Complement activity modulator

Abstract

To provide compositions comprising polypeptides that are useful as modulators of complement activity, and compositions for utilizing these modulators as therapeutics.SOLUTION: A pharmaceutical composition of the present disclosure is a water soluble pharmaceutical composition comprising: a) a C5 inhibitor; b) sodium chloride; and c) sodium phosphate, where the C5 inhibitor comprises a sequence Lys-Val-Glu-Arg-Phe-Asp-Xaa1-Xaa2-Tyr-Xaa3-Glu-Tyr-Pro-Xaa4-Lys, or a sequence having at least 90% identity thereto, where Xaa1 is N-methyl-aspartic acid, Xaa2 is tert-butylglycine, Xaa3 is 7-azatryptophan, and Xaa4 is cyclohexylglycine.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This invention relates to polypeptide-containing compounds that are useful as modulators of complement activity. Furthermore, methods for utilizing these modulators as therapeutic agents are also provided. [Background technology]

[0002] The immune response in vertebrates consists of adaptive and innate immune components. Adaptive immune responses are selective and slow-acting against specific pathogens, while components of innate immune responses recognize a wide range of pathogens and react rapidly upon infection. One such component of innate immune responses is the complement system.

[0003] The complement system contains approximately 20 circulating complement component proteins, primarily synthesized by the liver. These specific immune response components were initially named "complement" because they were observed to complement the antibody response in the destruction of bacteria. These proteins remain inactive until they are activated in response to infection. Activation occurs via a pathway of proteolytic cleavage initiated by pathogen recognition, leading to pathogen destruction. Three such pathways are known in the complement system: the classical pathway, the lectin pathway, and the alternative pathway. The classical pathway is activated when IgG or IgM molecules bind to the surface of a pathogen. The lectin pathway is initiated by mannan-binding lectin proteins that recognize sugar residues in the bacterial cell wall. The alternative pathway remains active at low levels even in the absence of any specific stimulus. While all three pathways differ in terms of their initiation events, all three converge on the cleavage of complement component C3. C3 is cleaved into two products, designated C3a and C3b. Of these, C3b is covalently bound to the pathogen surface, while C3a acts as a diffusive signal for promoting inflammation and mobilizing circulating immune cells. Surface-associated C3b forms complexes with other components to initiate cascade reactions between later components of the complement system. Because surface binding is required, complement activity maintains its localization and minimizes the destruction of non-target cells.

[0004] Pathogen-associated C3b promotes pathogen destruction in two ways. In one pathway, C3b is directly recognized by phagocytic cells, leading to phagocytosis of the pathogen. In the second pathway, pathogen-associated C3b initiates the formation of a membrane invasion complex (MAC). In the first step, C3b complexes with other complement components to form a C5 convertase complex. Depending on the initial complement activation pathway, the components of this complex may differ. C5 convertase formed as a result of the classical complement pathway contains C4b and C2a in addition to C3b. When formed by an alternative pathway, C5 convertase contains two subunits of C3b and one Bb component.

[0005] Complement component C5 is cleaved into C5a and C5b by either C5 convertase complex. C5a, much like C3a, diffuses into the circulatory system to promote inflammation and acts as a chemotactic for inflammatory cells. C5b remains bound to the cell surface and triggers MAC formation through interactions with C6, C7, C8, and C9. MACs are hydrophilic pores present throughout the cell membrane that disrupt cells by promoting the free flow of fluids into and out of the cell. [Overview of the project] [Problems that the invention aims to solve]

[0006] A crucial element of all immune activity is the immune system's ability to distinguish between self and non-self cells. Pathological conditions arise when the immune system fails to perform this distinction. In the complement system, vertebrate cells express proteins that protect themselves from the effects of the complement cascade. This ensures that the complement system's targets are limited to pathogenic cells. Many complement-related disorders and diseases involve the abnormal destruction of self-cells by the complement cascade. For example, subjects with paroxysmal nocturnal hemoglobinuria (PNH) are unable to synthesize functional complement regulatory proteins CD55 and CD59 on hematopoietic stem cells. This results in complement-mediated hemolysis and various downstream complications. Other complement-related disorders and diseases include, but are not limited to, autoimmune, neurological, hematological, and infectious diseases and disorders. Experimental evidence suggests that many complement-related disorders can be mitigated through the inhibition of complement activity. Therefore, compositions and methods for selectively blocking complement-mediated cell destruction are needed to treat these conditions. The present invention satisfies this need by providing related compositions and methods. [Means for solving the problem]

[0007] Summary of the Invention In some embodiments, the disclosure provides a pharmaceutical composition comprising R5000 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises sodium chloride at a concentration of about 25 mM to about 100 mM and sodium phosphate at a concentration of about 10 mM to about 100 mM. R5000 may be present at a concentration of about 1 mg / mL to about 400 mg / mL. The pharmaceutical composition may have a pH of about 6.5 to about 7.5. R5000 has an equilibrium dissociation constant (K) of about 0.1 nM to about 1 nM at C5. D ) may be bound. R5000 may block C5 production following activation of an alternative pathway for complement activation. R5000 may block the formation of membrane invasion complexes (MACs) following activation of the classical, alternative, or lectin pathways for complement activation.

[0008] In some embodiments, the disclosure provides a method for inhibiting hemolysis in a subject, comprising administering a pharmaceutical composition comprising R5000 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises sodium chloride at a concentration of about 25 mM to about 100 mM and sodium phosphate at a concentration of about 10 mM to about 100 mM. The pharmaceutical composition may be administered in a dose sufficient to achieve a plasma level of R5000 of about 0.1 μg / mL to about 20 μg / mL. Hemolysis may be inhibited by about 25% to 100% after administration. The pharmaceutical composition may be administered daily for at least 2 days. The pharmaceutical composition may be administered daily for 7 days. The pharmaceutical composition may be administered daily for at least 100 days. According to some methods, no adverse cardiovascular, respiratory, and / or central nervous system (CNS) effects are observed for at least 1 month after administration. According to some methods, no changes in heart rate and / or arterial pressure are observed for at least 1 month after administration. According to some methods, no changes in respiratory rate, tidal volume, and / or minute ventilation are observed for at least one month after administration.

[0009] In some embodiments, the Disclosure provides a method for inhibiting hemolysis in a subject, comprising administering a pharmaceutical composition comprising R5000 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises sodium chloride at a concentration of about 25 mM to about 100 mM and sodium phosphate at a concentration of about 10 mM to about 100 mM, wherein the pharmaceutical composition may be administered subcutaneously (SC) or intravenously (IV). The half-life (t) of R5000 levels in the subject plasma. 1 / 2 ) may be at least 4 hours. The R5000 level in the target plasma 1 / 2 The T of R5000 in the target plasma may be approximately 1 to 10 days. The steady-state distribution volume of R5000 in the target plasma may be approximately 10 mL / kg to approximately 200 mL / kg. The steady-state distribution volume of R5000 in the target plasma may correspond to at least 50% of the total blood volume. The total clearance rate of R5000 in the target plasma may be approximately 0.04 mL / hour / kg to approximately 4 mL / hour / kg. maxThe timeframe may be approximately 1 hour to approximately 48 hours. The presence of a measurable amount of R5000 may be substantially limited to the plasma compartment. The pharmaceutical composition may be administered in a dose sufficient to deliver approximately 0.01 mg to approximately 2 mg / kg of R5000 per subject body weight. Approximately 50% to approximately 99% of C5 activation in the subject may be inhibited. The pharmaceutical composition may be administered in a dose sufficient to deliver approximately 0.1 mg to approximately 0.4 mg / kg of R5000 per subject body weight. The pharmaceutical composition may be administered subcutaneously or intravenously. The pharmaceutical composition may be administered once or more times daily. The pharmaceutical composition may be administered over a period of 7 days. Percentage inhibition of hemolysis may be at least 90% to approximately 95% or more 3 hours after the first dose. Percentage inhibition of hemolysis may be at least 90% to approximately 95% or more as measured for at least 7 days after administration. Percentage inhibition of hemolysis may be at least 90% to approximately 95% or more for at least 4 days after administration. Maximum inhibition of hemolysis and / or maximum inhibition of complement activity may be achieved approximately 2 to 4 hours after administration.

[0010] In some embodiments, the disclosure provides a method for inhibiting hemolysis in a subject, comprising administering a pharmaceutical composition comprising R5000 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises sodium chloride at a concentration of about 25 mM to about 100 mM and sodium phosphate at a concentration of about 10 mM to about 100 mM, wherein R5000 is administered at a dose of 0.2 mg / kg. Hemolysis may be ≤3% 24 hours after the last dose. Complement activity may decrease from about 1 percent to about 10 percent over 7 days. Complement activity may be ≤5% 24 hours after the last dose. The pharmaceutical composition may be administered daily by subcutaneous or intravenous injection in a dose sufficient to deliver about 0.1 mg / day to about 60 mg / day / kg of subject body weight of R5000. The highest serum concentration (C) achieved max The concentration may be approximately 0.1 μg / mL to approximately 1000 μg / mL. The area under the curve (AUC) is approximately 200 μg. * time / mL ~ approx. 10,000μg * Time / mL may also be acceptable.

[0011] In some embodiments, the disclosure provides a method for treating paroxysmal nocturnal hemoglobinuria (PNH) in a subject requiring such treatment, comprising subcutaneous or intravenous administration of a pharmaceutical composition comprising R5000 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises sodium chloride at a concentration of about 25 mM to about 100 mM and sodium phosphate at a concentration of about 10 mM to about 100 mM. The subject may have been previously treated with an antibody-based therapeutic agent. The PNH in the subject may be resistant or unresponsive to treatment with an antibody-based therapeutic agent. The antibody-based therapeutic agent may be eculizumab.

[0012] In some embodiments, the Disclosure provides a kit comprising a pharmaceutical composition comprising R5000 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises sodium chloride at a concentration of about 25 mM to about 100 mM and sodium phosphate at a concentration of about 10 mM to about 100 mM.

[0013] In some embodiments, the Disclosure provides an automated injector comprising pharmaceutically acceptable excipients, wherein the pharmaceutically acceptable excipients include sodium chloride at concentrations of about 25 mM to about 100 mM and sodium phosphate at concentrations of about 10 mM to about 100 mM.

[0014] Other objectives, features, and advantages described above will become apparent from the following description of specific embodiments of the present invention, as well as from the attached drawings illustrating the principles of various embodiments of the present invention. [Brief explanation of the drawing]

[0015] [Figure 1] Scatter plot showing R5000 inhibition of C5a production. [Figure 2] Scatter plot showing R5000 inhibition of membrane invasion complex formation. [Figure 3] Scatter plot showing R5000 inhibitor activity in a cynomolgus monkey model. [Figure 4A]Scatter plot showing the pharmacokinetics and pharmacodynamic correlations of R5000 in male cynomolgus monkeys after multiple subcutaneous administrations at 0.21 mg / kg. [Figure 4B] Scatter plot showing the pharmacokinetics and pharmacodynamic correlations of R5000 in male cynomolgus monkeys after multiple subcutaneous administrations at 4.2 mg / kg. [Figure 5A] Graph showing R5000 levels over time after subcutaneous administration in rats and monkeys. [Figure 5B] Graph showing plasma concentrations over time after multiple subcutaneous doses of 0.21 and 4.2 mg / kg in monkeys. [Figure 6] A graph showing the predicted plasma concentration of R5000 in humans with daily administration of R5000. [Figure 7] A line graph showing the concentration of R5000 in cynomolgus monkeys after the first dose in a repeated-dose toxicity study. [Figure 8] A line graph showing the concentration of R5000 in cynomolgus monkeys after the final dose in a repeated-dose toxicity study. [Figure 9A] A graph showing the change in percentage hemolysis relative to R5000 concentration in a human study involving multiple doses. [Figure 9B] A graph showing the time course of plasma concentrations of R5000 in a multi-dose human study. [Figure 10A] A graph showing the time course of complement activity induced by R5000 treatment in a human trial involving multiple doses. [Figure 10B] A graph showing long-term changes in complement activity after R5000 treatment in a human trial involving multiple doses. [Figure 11A] A graph showing the dose-dependent peak plasma concentration levels of R5000 in a single-dose escalation clinical trial in humans. [Figure 11B] A graph showing the time course of plasma concentrations after a single dose of R5000. [Figure 12A] A graph showing the percentage of hemolysis over time following a single dose of R5000 administered to humans over a 4-day period. [Figure 12B]A graph showing the percentage CH50 over time after a single dose of R5000 in humans. [Figure 12C] A graph showing percentage hemolysis at various doses over a 28-day period in humans. [Figure 13] A graph showing the percentage complement activity over time after a single dose of R5000 in humans. [Modes for carrying out the invention]

[0016] Detailed explanation I. Compounds and Compositions The present invention provides compounds and compositions that function to modulate complement activity. Such compounds and compositions of the present invention may include inhibitors that block complement activation. As used herein, “complement activity” includes activation of the complement cascade, formation of cleavage products from complement components such as C3 or C5, assembly of downstream complexes after a cleavage event, or any process or event that accompanies or results from the cleavage of complement components such as C3 or C5. Complement inhibitors may include C5 inhibitors that block complement activation at the level of complement component C5. C5 inhibitors can bind to C5 and prevent its cleavage by C5 convertase into cleavage products C5a and C5b. As used herein, “complement component C5” or “C5” is defined as a complex that is cleaved by C5 convertase into at least cleavage products C5a and C5b. “C5 inhibitor” as used herein includes any compound or composition that inhibits the processing or cleavage of the complement component C5 complex or the cleavage products of complement component C5 before cleavage.

[0017] It is understood that inhibition of C5 cleavage prevents the assembly and activity of cytolytic membrane invasion complexes (MACs) against glycosylphosphatidylinositol (GPI) adhesion protein-deficient erythrocytes. Therefore, in some cases, the C5 inhibitors of the present invention may also bind to C5b and prevent C6 binding and subsequent assembly of the C5b-9 MAC.

[0018] Peptide-based compounds In some embodiments, the C5 inhibitors of the present invention are polypeptides. According to the present invention, any amino acid-based molecule (natural or unnatural) may be referred to as a "polypeptide," a term that encompasses "peptides," "peptidomimetics," and "proteins." A "peptide" is traditionally considered to be in the size range of about 4 to about 50 amino acids. Polypeptides larger than about 50 amino acids are generally referred to as "proteins."

[0019] C5 inhibitor polypeptides may be linear or cyclic. Cyclic polypeptides include any polypeptide having one or more cyclic features, such as loops and / or internal bonds, as part of their structure. In some embodiments, a cyclic polypeptide is formed when a molecule acts as a crosslinking portion to link two or more regions of a polypeptide. As used herein, the term “crosslinking portion” refers to one or more components of a crosslink formed between two adjacent or non-adjacent amino acids, unnatural amino acids, or non-amino acids in a polypeptide. The crosslinking portion may be a composition of any size. In some embodiments, the crosslinking portion may include one or more chemical bonds between two adjacent or non-adjacent amino acids, unnatural amino acids, non-amino acid residues, or a combination thereof. In some embodiments, such chemical bonds may be present between one or more functional groups in adjacent or non-adjacent amino acids, unnatural amino acids, non-amino acid residues, or a combination thereof. The crosslinking portion may include one or more amide bonds (lactams), disulfide bonds, thioether bonds, aromatic rings, triazole rings, and hydrocarbon chains. In some embodiments, the crosslinking portion includes an amide bond between amine functionality and carboxylic acid functionality (each present in the side chain of an amino acid, unnatural amino acid, or non-amino acid residue). In some embodiments, the amine or carboxylic acid functionality is part of a non-amino acid residue or a non-natural amino acid residue.

[0020] The C5 inhibitor polypeptide may be cyclized via the carboxyl terminus, the amino terminus, or any other convenient attachment site, such as the sulfur of cysteine ​​(e.g., via the formation of a disulfide bond between two cysteine ​​residues in the sequence), or via any side chain of an amino acid residue. Furthermore, the bonds forming the cyclic loop may include, but are not limited to, maleimide bonds, amide bonds, ester bonds, ether bonds, thiol ether bonds, hydrazone bonds, or acetamide bonds.

[0021] In some embodiments, the cyclic C5 inhibitor polypeptide of the present invention is formed using a lactam moiety. Such cyclic polypeptides may be formed, for example, by synthesis on a Wang resin solid support using standard Fmoc chemistry. In some cases, Fmoc-ASP(allyl)-OH and Fmoc-LYS(aloc)-OH are incorporated into the polypeptide to serve as precursor monomers for lactam crosslinking.

[0022] The C5 inhibitor polypeptide of the present invention may be peptidomimetic. A "peptidomimetic" or "polypeptidomimetic" is a polypeptide in which the molecule contains structural elements not found in natural polypeptides (i.e., polypeptides composed only of the 20 protein-constituting amino acids). In some embodiments, peptidomimetic molecules can reproduce or mimic the biological effects of natural peptides. Peptidomimetic molecules may differ from natural polypeptides in many respects, for example, through changes in their skeletal structure or through the presence of amino acids that do not exist in nature. In some cases, peptidomimetic amino acids may include side chains not found in the 20 known protein-constituting amino acids; non-polypeptide crosslinking moieties used to carry out cyclization between the ends or within the molecule; substitution of amide bond hydrogens with methyl groups (N-methylation) or other alkyl groups; exchange of peptide bonds with chemical groups or chemical bonds resistant to chemical or enzymatic treatment; N-terminus and C-terminus modifications; and / or amino acids having conjugations with non-peptide extensions (e.g., polyethylene glycol, lipids, carbohydrates, nucleosides, nucleotides, nucleoside bases, various small molecules, or phosphate or sulfate groups).

[0023] As used herein, the term “amino acid” includes residues of both natural and non-natural amino acids. The 20 natural protein-forming amino acids are identified and referenced herein by one-letter or three-letter names such as aspartic acid (Asp:D), isoleucine (Ile:I), threonine (Thr:T), leucine (Leu:L), serine (Ser:S), tyrosine (Tyr:Y), glutamic acid (Glu:E), phenylalanine (Phe:F), proline (Pro:P), histidine (His:H), glycine (Gly:G), lysine (Lys:K), alanine (Ala:A), arginine (Arg:R), cysteine ​​(Cys:C), tryptophan (Trp:W), valine (Val:V), glutamine (Gln:Q), methionine (Met:M), and asparagine (Asn:N). Natural amino acids exist in their levorotatory (L) stereoisomers. Amino acids as referred herein are L-stereoisomers unless otherwise specified. The term “amino acid” also includes amino acids having conventional amino protecting groups (e.g., acetyl or benzyloxycarbonyl), as well as natural and unnatural amino acids with protected carboxyl termini (e.g., as (C1-C6) alkyl, phenyl, or benzyl esters or amides; or as α-methylbenzylamides). Other suitable amino and carboxyl protecting groups are known to those skilled in the art (see, for example, Greene, TW; Wutz, PGM, “Protecting Groups In Organic Synthesis,” 2nd edition, 1991, John Wiley & Sons, Inc., New York, and the documents cited therein (the contents of each of these are incorporated herein by reference in their entirety)). Polypeptides and / or polypeptide compositions of the present invention may also contain modified amino acids.

[0024] "Non-natural" amino acids have side chains or other features not present in the 20 natural amino acids listed above, and include, but are not limited to, N-methyl amino acids, N-alkyl amino acids, α,α-substituted amino acids, β-amino acids, α-hydroxy amino acids, D-amino acids, and other non-natural amino acids known in the art (e.g., Josephson et al., 2005, Journal of the American Chemical Society (J.Am.C.Soc.), Vol. 127). See also pp. 11727-11735; Forster, AC et al., 2003, Proceedings of the National Academy of Sciences (Proc. Natl. Acad. Sci. USA), Vol. 100, pp. 6353-6357; Subtelny et al., 2008, Journal of the American Chemical Society (J. Am. Chem. Soc.), Vol. 130, pp. 6131-6136; Hartman, MCT et al., 2007, PLoS ONE, 2:e972; and Hartman et al., 2006, Proceedings of the National Academy of Sciences (Proc. Natl. Acad. Sci. USA), Vol. 103, pp. 4356-4361). Further non-natural amino acids useful for optimizing polypeptides and / or polypeptide compositions according to the present invention include 1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid, 1-amino-2,3-hydro-1H-indene-1-carboxylic acid, homolysine, homoarginine, homoserine, 2-aminoadipic acid, 3-aminoadipic acid, β-alanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 5-aminopentanoic acid, 5-aminohexanoic acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, desmosine, 2,3-Diaminopropionic acid, N-ethylglycine, N-ethylasparagine, homoproline, hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylpentylglycine, naphthylalanine, ornithine, pentylglycine, thioproline, norvaline, tert-butylglycine, phenylglycine, azatryptophan, 5-azatryptophan, 7-azatryptophan, 4-fluorophenylalanine, penicillamine, sarcosine, homocysteine, 1-aminocyclopropanecarboxylic acid, 1-aminocyclobutanecarboxylic acid, 1-aminocyclopentanecarboxylic acid, 1-aminocyclohexanecarboxylic acid, 4-aminotetrahydro-2H-pyran-4-carboxylic acid, (S)-2-amino-3-(1H-tetrazoleucine) Propanoic acid, cyclopentylglycine, cyclohexylglycine, cyclopropylglycine, η-ω-methylarginine, 4-chlorophenylalanine, 3-chlorotyrosine, 3-fluorotyrosine, 5-fluorotryptophan, 5-chlorotryptophan, citrulline, 4-chloro-homophenylalanine, homophenylalanine, 4-aminomethylphenylalanine, 3-aminomethylphenylalanine, octylglycine, norleucine, tranexamic acid, 2-aminopentanoic acid, 2-aminohexanoic acid, 2-aminoheptanoic acid, 2-aminooctanoic acid, 2-aminononanoic acid, 2-aminodecanoic acid, 2-aminoundecanoic acid, 2-aminododecanoic acid, aminovaleric acid and 2-(2-aminoethoxy)acetic acid, pipecoric acid, 2-carboxyazetidine, hexafluoroleucine, 3-fluorovaline, 2-amino-4,4-Difluoro-3-methylbutanoic acid, 3-fluoroisoleucine, 4-fluoroisoleucine, 5-fluoroisoleucine, 4-methylphenylglycine, 4-ethylphenylglycine, 4-isopropylphenylglycine, (S)-2-amino-5-azidopentaic acid (also referred to herein as "X02"), (S)-2-aminohepto-6-enoic acid (also referred to herein as "X30"), (S)-2-aminopento-4-ic acid (also referred to herein as " (Also referred to as "X31"), (S)-2-aminopento-4-enoic acid (also referred to herein as "X12"), (S)-2-amino-5-(3-methylguanidino)pentanoic acid, (S)-2-amino-3-(4-(aminomethyl)phenyl)propanoic acid, (S)-2-amino-3-(3-(aminomethyl)phenyl)propanoic acid, (S)-2-amino-4-(2-aminobenzo[d]oxazole-5-yl)butanoic acid, (S)-leucinol, (S)-valinol, ( S)-tert-leucinol, (R)-3-hetilbutan-2-amine, (S)-2-methyl-1-phenylpropan-1-amine, and (S)-N,2-dimethyl-1-(pyridine-2-yl)propan-1-amine, (S)-2-amino-3-(oxazole-2-yl)propanoic acid, (S)-2-amino-3-(oxazole-5-yl)propanoic acid, (S)-2-amino-3-(1,3,4-oxadiazole-2-yl)propanoic acid, (S)-2-amino-3-(1, 2,4-Oxadiazole-3-yl)propanoic acid, (S)-2-amino-3-(5-fluoro-1H-indazole-3-yl)propanoic acid, and (S)-2-amino-3-(1H-indazole-3-yl)propanoic acid, (S)-2-amino-3-(oxazol-2-yl)butanoic acid, (S)-2-amino-3-(oxazol-5-yl)butanoic acid, (S)-2-amino-3-(1,3,4-oxadiazole-2-yl)butanoic acid, (S)-2-amino-3-(1,2,Examples include, but are not limited to, 4-oxadiazole-3-yl)butanoic acid, (S)-2-amino-3-(5-fluoro-1H-indazole-3-yl)butanoic acid, and (S)-2-amino-3-(1H-indazole-3-yl)butanoic acid, 2-(2'MeOphenyl)-2-aminoacetic acid, tetrahydro-3-isoquinoline carboxylic acid, and their stereoisomers (including, but not limited to, D and L isomers).

[0025] Other non-natural amino acids useful for optimizing polypeptides or polypeptide compositions according to the present invention include, but are not limited to, fluorinated amino acids in which one or more carbon-bonded hydrogen atoms are replaced by fluorine. The number of fluorine atoms included can range from one to all hydrogen atoms. Examples of such amino acids include, but are not limited to, 3-fluoroproline, 3,3-difluoroproline, 4-fluoroproline, 4,4-difluoroproline, 3,4-difluoroproline, 3,3,4,4-tetrafluoroproline, 4-fluorotryptophan, 5-fluorotryptophan, 6-fluorotryptophan, 7-fluorotryptophan, and their stereoisomers.

[0026] Further non-natural amino acids useful for optimizing polypeptides according to the present invention include, but are not limited to, those with two α-carbon substitutions. These include amino acids in which the two substituents on the α-carbon are the same, such as α-aminoisobutyric acid and 2-amino-2-ethylbutanoic acid, as well as those with different substituents, such as α-methylphenylglycine and α-methylproline. Furthermore, substituents on the α-carbon may form a ring together, for example, 1-aminocyclopentanecarboxylic acid, 1-aminocyclobutanecarboxylic acid, 1-aminocyclohexanecarboxylic acid, 3-aminotetrahydrofuran-3-carboxylic acid, 3-aminotetrahydropyran-3-carboxylic acid, 4-aminotetrahydropyran-4-carboxylic acid, 3-aminopyrrolidine-3-carboxylic acid, 3-aminopiperidine-3-carboxylic acid, 4-aminopiperidine-4-carboxylic acid, and their stereoisomers.

[0027] Other non-natural amino acids useful for optimizing polypeptides or polypeptide compositions according to the present invention include, but are not limited to, analogs of tryptophan in which the indole ring system is replaced by another 9 or 10-membered bicyclic ring system containing 0, 1, 2, 3, or 4 heteroatoms independently selected from N, O, or S. Each ring system may be saturated, partially unsaturated, or fully unsaturated. Any of the substituted atoms in the ring system may be substituted by 0, 1, 2, 3, or 4 substituents. Each substituent may be independently selected from H, F, Cl, Br, CN, COOR, CONRR', oxo, OR, and NRR'. Each R and R' may be independently selected from H, C1-C20 alkyl, or C1-C20 alkyl-O-C1-20 alkyl.

[0028] In some embodiments, tryptophan analogs (also referred to herein as “tryptophan analogs”) may be useful in optimizing the polypeptides or polypeptide compositions of the present invention. Tryptophan analogs may include, but are not limited to, 5-fluorotryptophan [(5-F)W], 5-methyl-O-tryptophan [(5-MeO)W], 1-methyltryptophan [(1-Me-W) or (1-Me)W], D-tryptophan (D-Trp), azatryptophan (including, but not limited to, 4-azatryptophan, 7-azatryptophan, and 5-azatryptophan), 5-chlorotryptophan, 4-fluorotryptophan, 6-fluorotryptophan, 7-fluorotryptophan, and their stereoisomers. Unless otherwise specified, the term “azatryptophan” and its abbreviation “azaTrp” refer to 7-azatryptophan as used herein.

[0029] Modified amino acid residues useful for optimizing polypeptides and / or polypeptide compositions according to the present invention include, but are not limited to, those that are chemically blocked (reversible or irreversible); those with a chemically modified N-terminal amino group or its side chain group; those with a chemically modified amide skeleton, such as N-methylated residues; D (non-natural amino acid) and L (natural amino acid) stereoisomers; or residues with chemically modified side chain functional groups that have become other functional groups. In some embodiments, modified amino acids include, but are not limited to, methionine sulfoxides, methionine sulfones, aspartate-(β-methyl ester), modified amino acids of aspartate, N-ethylglycine, modified amino acids of glycine, alanine carboxamide, and / or modified amino acids of alanine. Non-natural amino acids can be purchased from Sigma-Aldrich (St. Louis, Missouri), Bachem (Trans, California), or other suppliers. Non-natural amino acids may further include any of those listed in Table 2 of U.S. Patent Application Publication No. 2011 / 0172126 (the contents of which are incorporated herein by reference in their entirety).

[0030] This invention examines variants and derivatives of polypeptides presented herein. These include variants and derivatives resulting from substitution, insertion, deletion, and covalent bonding. As used herein, the term “derivative” is used synonymously with the term “variant” and refers to a molecule that has been modified or altered to some extent with respect to the reference molecule or initiation molecule.

[0031] The polypeptides of the present invention may contain any of the following components, features, or parts, if the abbreviations used herein include: "Ac" and "NH2" represent acetyl and amidated ends, respectively; "Nvl" is an abbreviation for norvaline; "Phg" is an abbreviation for phenylglycine; "Tbg" is an abbreviation for tert-butylglycine; "Chg" is an abbreviation for cyclohexylglycine; and "(N-Me)X" is an abbreviation for the N-methylated form of an amino acid represented by a one- or three-letter amino acid code in place of the variable "X" written as N-methyl-X [for example, (N-Me)D or (N-Me)Asp is as "AzaTrp" is an abbreviation for azatryptophan, "(4-F)Phe" is an abbreviation for 4-fluorophenylalanine, "Tyr(OMe)" is an abbreviation for O-methyltyrosine, "Aib" is an abbreviation for aminoisobutyric acid, "(homo)F" or "(homo)Phe" is an abbreviation for homophenylalanine, "(2-OMe)Phg" means 2-O-methylphenylglycine, and "(5-F)W" means 5-fluorotryptophan. The abbreviations mean that "DX" means the D-stereoisomer of a given amino acid "X" [for example, (D-Chg) is an abbreviation for D-cyclohexylglycine], "(5-MeO)W" means 5-methyl-O-tryptophan, "HomoC" means homocysteine, "(1-Me-W)" or "(1-Me)W" means 1-methyltryptophan, "Nle" means norleucine, "Tiq" means tetrahydroisoquinoline residue, and "Asp(T)" means (S)-2-amino-3-(1H-tetra "(3-Cl-Phe)" means rhazol-5-yl)propanoic acid, "[(N-Me-4-F)Phe]" or "[(N-Me-4-F)Phe]" means N-methyl-4-fluorophenylalanine, "(m-Cl-homo)Phe" means meta-chlorohomophenylalanine, "(des-amino)C" means 3-thiopropionic acid, "(α-methyl)D" means α-methyl-L-aspartic acid, and "2Nal" means 2-naphthylalanine."(3-aminomethyl)Phe" means 3-aminomethyl-L-phenyalanine, "Cle" means cycloleucine, "Ac-pyran" means 4-amino-tetrahydropyran-4-carboxylic acid, "(Lys-C16)" means N-ε-palmitoyl lysine, "(Lys-C12)" means N-ε-lauryl lysine, "(Lys-C10)" means N-ε-caprylyl lysine, "(Lys-C8)" means N-ε-caprylic acid lysine, "[xxylyl(y,z)]" means xylyl crosslinking between two thiol-containing amino acids, where x can be m, p, or o, and indicates the use of (respectively) meta-, para-, or ortho-dibromoxylen to form the crosslinking, and the numerical identifiers y and z are rings The amino acid positions within the polypeptide of amino acids involved in the formation are determined, "[cyclo(y,z)]" means the formation of a bond between two amino acid residues, where the numerical identifiers y and z determine the positions of the residues involved in the bond, "[cyclo-olefinyl(y,z)]" means the formation of a bond between two amino acid residues by olefin metathesis, where the numerical identifiers y and z determine the positions of the residues involved in the bond, "[cyclo-thioalkyl(y,z)]" means the formation of a thioether bond between two amino acid residues, where the numerical identifiers y and z determine the positions of the residues involved in the bond, and "[cyclo-triazolyl(y,z)]" means the formation of a triazole ring between two amino acid residues, where the numerical identifiers y and z determine the positions of the residues involved in the bond. "B20" stands for N-ε-(PEG2-γ-glutamic acid-N-α-octadecanediic acid)lysine [also known as (1S,28S)-1-amino-7,16,25,30-tetraoxo-9,12,18,21-tetraoxa-6,15,24,29-tetraazahexatetracontane-1,28,46-tricarboxylic acid].

[0032] B20

[0033] [ka]

[0034] "B28" stands for N-ε-(PEG24-γ-glutamic acid-N-α-hexadecanoyl)lysine. B28

[0035] [ka]

[0036] "K14" stands for N-ε-1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl--L-lysine. All other symbols represent standard single-letter amino acid codes.

[0037] Some C5 inhibitor polypeptides contain approximately 5 to 10 amino acids, 6 to 12 amino acids, 7 to 14 amino acids, 8 to 16 amino acids, 10 to 18 amino acids, 12 to 24 amino acids, or 15 to 30 amino acids. In some cases, C5 inhibitor polypeptides contain at least 30 amino acids.

[0038] Some C5 inhibitors of the present invention include a C-terminal lipid moiety. Such a lipid moiety may include a fatty acid acyl group (for example, a saturated or unsaturated fatty acid acyl group). In some cases, the fatty acid acyl group may be a palmitoyl group.

[0039] A C5 inhibitor having a fatty acid acyl group may contain one or more molecular linkers that link the fatty acid to a peptide. Such molecular linkers may contain amino acid residues. In some cases, an L-γ-glutamic acid residue may be used as the molecular linker. In some cases, the molecular linker may contain one or more polyethylene glycol (PEG) linkers. The PEG linker of the present invention may contain about 1 to about 5, about 2 to about 10, about 4 to about 20, about 6 to about 24, about 8 to about 32, or at least 32 PEG units.

[0040] The C5 inhibitor of the present invention may have a molecular weight of approximately 200 g / mol to approximately 600 g / mol, approximately 500 g / mol to approximately 2000 g / mol, approximately 1000 g / mol to approximately 5000 g / mol, approximately 3000 g / mol to approximately 4000 g / mol, approximately 2500 g / mol to approximately 7500 g / mol, approximately 5000 g / mol to approximately 10000 g / mol, or at least 10000 g / mol.

[0041] In some embodiments, the C5 inhibitor polypeptide of the present invention comprises R5000. The core amino acid sequence of R5000 (SEQ ID NO: 1) comprises 15 amino acids (all L-amino acids) including four non-natural amino acids (N-methyl-aspartic acid, tert-butylglycine, 7-azatryptophan, and cyclohexylglycine); a lactam crosslink between K1 and D6 of the polypeptide sequence; and a C-terminal lysine residue having a modified side chain that forms an N-ε-(PEG24-γ-glutamic acid-N-α-hexadecanoyl)lysine residue (also referred to herein as "B28"). The C-terminal lysine side chain modification comprises a polyethylene glycol (PEG) spacer (PEG24), where PEG24 is attached to an L-γ-glutamic acid residue derivatized with a palmitoyl group.

[0042] In some embodiments, the present invention includes variants of R5000. In some R5000 variants, the C-terminal lysine side chain portion may be modified. In some cases, the PEG24 spacer (having a 24PEG subunit) of the C-terminal lysine side chain portion may contain fewer or additional PEG subunits. In other cases, the palmitoyl group of the C-terminal lysine side chain portion may be substituted with another saturated or unsaturated fatty acid. Further, the L-γ-glutamic acid linker of the C-terminal lysine side chain portion (between the PEG and acyl groups) may be substituted with an alternative amino acid or a non-amino acid linker.

[0043] In some embodiments, the R5000 variant may include modifications to the core polypeptide sequence in R5000, which may be combined with features of the cyclic or C-terminal lysine side chain portion of R5000. Such variants may have at least 50%, at least 55%, at least 65%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with respect to the core polypeptide sequence of SEQ ID NO: 1. In some cases, the R5000 variant may be cyclized between amino acids other than those used in R5000 by forming lactam crosslinks.

[0044] The C5 inhibitors of the present invention may be developed or modified to achieve specific binding properties. Inhibitor binding may be evaluated by measuring the ratio of association and / or dissociation with a particular target. In some cases, the compound exhibits potent and rapid association with the target combined with slow dissociation. In some embodiments, the C5 inhibitors of the present invention exhibit potent and rapid association with C5. Such inhibitors may further exhibit slow dissociation with C5.

[0045] The C5 inhibitors of the present invention that bind to the C5 complement protein are approximately 0.001 nM to approximately 0.01 nM, approximately 0.005 nM to approximately 0.05 nM, approximately 0.01 nM to approximately 0.1 nM, approximately 0.05 nM to approximately 0.5 nM, approximately 0.1 nM to approximately 1.0 nM, approximately 0.5 nM to approximately 5.0 nM, approximately 2 nM to approximately 10 nM, and approximately 8 nM to approximately Equilibrium dissociation constants (K) of 20 nM, approximately 15 nM to 45 nM, approximately 30 nM to 60 nM, approximately 40 nM to 80 nM, approximately 50 nM to 100 nM, approximately 75 nM to 150 nM, approximately 100 nM to 500 nM, approximately 200 nM to 800 nM, approximately 400 nM to 1,000 nM, or at least 1,000 nM. D ) may bind to the C5 complement protein.

[0046] In some embodiments, the C5 inhibitors of the present invention block the formation or production of C5a from C5. In some cases, the formation or production of C5a is blocked following the activation of an alternative pathway for complement activation. In some cases, the C5 inhibitors of the present invention block the formation of membrane invasion complexes (MACs). Such inhibition of MAC formation may be due to the binding of the C5 inhibitor to the C5b subunit. The binding of the C5 inhibitor to the C5b subunit may prevent the binding of C6, resulting in interference with MAC formation. In some embodiments, this inhibition of MAC formation occurs after the activation of a classical, alternative, or lectin pathway.

[0047] The C5 inhibitors of the present invention may be synthesized using chemical processes. In some cases, such synthesis eliminates the risks associated with the production of biological products in mammalian cell lines. In some cases, chemical synthesis may be simpler and more cost-effective than biological production processes.

[0048] Isotope mutations Polypeptides according to the present invention may contain one or more atoms that are isotopes. As used herein, the term “isotope” means a chemical element having one or more additional neutrons. In one embodiment, polypeptides according to the present invention may be deuterized. As used herein, the term “deuterized” means a substance having one or more hydrogen atoms replaced by a deuterium isotope. A deuterium isotope is an isotope of hydrogen. While a hydrogen nucleus contains one proton, a deuterium nucleus contains both a proton and a neutron. Compounds and pharmaceutical compositions according to the present invention may be deuterized to alter their physical properties, such as stability, or to make them usable for diagnostic and experimental purposes.

[0049] II.How to use Provided herein are methods for regulating complement activity using the compounds and / or compositions of the present invention.

[0050] treatment index A crucial component of all immune activity (innate and adaptive) is the immune system's ability to distinguish between self and non-self cells. Pathogenesis occurs when the immune system is unable to make this distinction. In the case of the complement system, vertebrate cells express inhibitory proteins that protect them from the effects of the complement cascade, ensuring that the complement system is specific to pathogenic microorganisms. Numerous complement-related disorders and diseases are associated with the abnormal destruction of self cells by the complement cascade.

[0051] The methods of the present invention include methods for treating complement-related disorders using the compounds and compositions of the present invention. When referred to herein, “complement-related disorders” may include any conditions relating to the cleavage or processing of complement components, such as C5, which are dysfunctions of the complement system.

[0052] In some embodiments, the methods of the present invention include methods for inhibiting complement activity in a subject. In some cases, the percentage of complement activity inhibited in the subject may be at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.9%. In some cases, this level of inhibition and / or maximum inhibition of complement activity may be achieved from about 1 hour to about 3 hours after administration, from about 2 hours to about 4 hours after administration, from about 3 hours to about 10 hours after administration, from about 5 hours to about 20 hours after administration, or from about 12 hours to about 24 hours after administration. Inhibition of complement activity may continue throughout a period of at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks. In some cases, this level of inhibition may be achieved through daily administration. Such daily administration may include administration for at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 2 months, at least 4 months, at least 6 months, at least 1 year, or at least 5 years. In some cases, a subject may be administered the compounds or compositions of the Disclosure for the life of such subject.

[0053] In some embodiments, the methods of the present invention include methods for inhibiting C5 activity in a subject. “C5-dependent complement activity” or “C5 activity,” as used herein, refers to activation of the complement cascade via C5 cleavage, assembly of downstream cleavage products of C5, or any other process or event associated with or resulting from C5 cleavage. In some cases, the percentage of C5 activity inhibited in a subject may be at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.9%.

[0054] In some embodiments, the methods of the present invention may include a method of inhibiting hemolysis by administering one or more compounds or compositions of the present invention to a subject or patient in need thereof. According to some such methods, hemolysis may be reduced to about 25% to about 99%. In other embodiments, hemolysis is reduced to about 10% to about 40%, about 25% to about 75%, about 30% to about 60%, about 50% to about 90%, about 75% to about 95%, about 90% to about 99%, or about 97% to about 99.5%. In some cases, hemolysis is reduced to at least 50%, 60%, 70%, 80%, 90%, or 95%.

[0055] According to some methods, the percentage inhibition of hemolysis is approximately ≥90% to ≥99% (e.g., ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%). In some cases, this level of inhibition and / or maximum inhibition of hemolysis may be achieved from approximately 1 hour to 3 hours after administration, 2 hours to 4 hours after administration, 3 hours to 10 hours after administration, 5 hours to 20 hours after administration, or 12 hours to 24 hours after administration. Inhibition of hemolytic activity levels may continue throughout a period of at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks. In some cases, this level of inhibition may be achieved through daily administration. Such daily administration may include administration for at least two days, at least three days, at least four days, at least five days, at least six days, at least seven days, at least two weeks, at least three weeks, at least four weeks, at least two months, at least four months, at least six months, at least one year, or at least five years. In some cases, the compounds or compositions of the Disclosure may be administered to a subject for the life of such subject.

[0056] The C5 inhibitors of the present invention may be used to treat one or more symptoms, with minimal or no adverse effects resulting from C5 inhibitor treatment. In some cases, no adverse cardiovascular, respiratory, and / or central nervous system (CNS) effects occur. In some cases, no changes in heart rate and / or arterial pressure occur. In some cases, no changes in respiratory rate, tidal volume, and / or minute ventilation occur.

[0057] "Reduction" or "decrease" in relation to disease markers or symptoms means a significant, often statistically significant, reduction at such levels. This reduction could be, for example, at least 10%, at least 20%, at least 30%, at least 40%, or more, and preferably to a level that is considered within the normal range in individuals without such disorder.

[0058] "Increasing" or "elevating" in relation to a disease marker or symptom means a significant, often statistically significant, increase at such a level. The increase could be, for example, at least 10%, at least 20%, at least 30%, at least 40%, or more, and preferably to a level that is considered within the normal range in individuals without such disorder.

[0059] A therapeutic or preventive effect is evident when there is a significant, often statistically significant, improvement in one or more parameters of the disease, or in preventing the worsening or development of symptoms that would normally be expected. For example, a favorable change of at least 10%, preferably at least 20%, 30%, 40%, 50%, or more, in a measurable parameter of the disease may indicate an effective treatment. The efficacy of a given compound or composition may also be determined using experimental animal models of a given disease known in the art. When using experimental animal models, the efficacy of the treatment is demonstrated when there is a statistically significant adjustment in a marker or symptom.

[0060] Paroxysmal nocturnal hemoglobinuria In some embodiments, provided herein are methods for treating paroxysmal nocturnal hemoglobinuria (PNH) using compounds or compositions of the present invention, such as pharmaceutical compositions. PNH is a rare complement-related disorder caused by acquired mutations in the phosphatidylinositol glycan anchor biosynthesis class A (PIG-A) gene derived from pluripotent hematopoietic stem cells (Pu, JJ et al., Clinical and Translational Science (Clin Transl Sci.), June 2011, Vol. 4, No. 3, pp. 219-24). PNH is characterized by bone marrow dysfunction, hemolytic anemia, and thrombosis. The PIG-A gene product is required for the production of glycosylphosphatidylinositol (GPI), a glycolipid anchor used to tether proteins to the plasma membrane. Two complement regulatory proteins involved in protecting cells from the lytic activity of terminal complement complex (PTF) CD55 (a decomposition promoter) and CD59 (a membrane inhibitor of reactive lysis) become non-functional in the absence of GPI. This leads to C5 activation and the accumulation of specific complement proteins on the surface of red blood cells (RBCs), resulting in complement-mediated disruption of these cells.

[0061] Patients with PNH initially present with hemoglobinuria, abdominal pain, smooth muscle dystonia, and fatigue, for example, PNH-related symptoms or disorders. PNH is also characterized by intravascular hemolysis (the primary clinical symptom of the disease) and venous thrombosis. Venous thrombosis can occur in unusual sites, including, but not limited to, hepatic veins, mesenteric veins, cerebral veins, and cutaneous veins (Parker, C. et al., 2005, Blood, Vol. 106, pp. 3699-709 and Parker, CJ, 2007, Experimental Hematol., Vol. 35, pp. 523-33). Currently, eculizumab (SOLIRIS®, Alexion Pharmaceuticals, Cheshire, Connecticut), a C5 inhibitor monoclonal antibody, is the only approved treatment for PNH.

[0062] Treatment with eculizumab provides adequate control of intravascular hemolysis in most PNH patients (Schrezenmeier, H. et al., 2014, Haematologica, Vol. 99, pp. 922-99). However, Nishimura and colleagues have described 11 patients in Japan (3.2% of patients with PNH) who had mutations in the C5 gene that inhibited eculizumab binding to C5 and were unresponsive to antibody treatment (Nishimura, JI. et al., 2014, New England Journal of Medicine, Vol. 370, pp. 632-639). Furthermore, eculizumab is administered every two weeks as an IV infusion under the supervision of a medical professional, which is inconvenient and burdensome for patients.

[0063] Long-term intravenous administration can lead to serious complications such as infection, local thrombosis, hematoma, and progressively reduced venous access. In addition, eculizumab is a large protein and is associated with an increased risk of immunogenicity and hypersensitivity. Ultimately, while eculizumab binds to C5 and inhibits C5b production, any C5b produced due to incomplete inhibition can initiate MAC formation, potentially leading to hemolysis.

[0064] Peripheral blood from patients with PNH can show a variability in the ratio of normal to abnormal cells. The disease is classified based on its clinical features, bone marrow characteristics, and the percentage of GPI-AP-deficient polymorphonuclear leukocytes (PMNs) by the International PNH Interest Committee. They are further classified according to the Group. Since GPI-AP-deficient erythrocytes are more susceptible to destruction in PNH patients, flow cytometry analysis of PMN is considered to provide more useful information (Parker, CJ, 2012, Current Opinion in Hematology, Vol. 19, pp. 141-148). Flow cytometry analysis in classical PNH shows 50-100% GPI-AP-deficient PMN.

[0065] Hemolytic anemia in PNH is independent of autoantibodies (Coombs-negative) and is caused by unregulated activation of the alternative complement pathway (AP). In some embodiments, the compounds and compositions of the present invention, such as pharmaceutical compositions, are particularly useful in the treatment of PNH. Such compounds and compositions may include C5 inhibitors (e.g., R5000). The C5 inhibitors of the present invention that are useful for the treatment of PNH can, in some cases, block the cleavage of C5 to C5a and C5b. In some cases, the C5 inhibitors of the present invention may be used as an alternative to eculizumab therapeutics for PNH. Unlike eculizumab, the C5 inhibitors of the present invention can bind to C5b, preventing binding to C6 and subsequent assembly of the C5b-9MAC.

[0066] In some cases, R5000 and its compositions may be used to treat PNH in subjects. Such subjects may include those that have had adverse reactions to, were unresponsive to, showed reduced responsiveness to, or shown resistance to other treatments (e.g., using eculizumab). In some embodiments, treatment using the compounds and compositions of this disclosure may inhibit hemolysis of PNH erythrocytes in a dose-dependent manner.

[0067] In some embodiments, R5000 is administered in combination with eculizumab in a regimen that may include parallel or sequential treatment. Based on sequence and structural data, R5000 may be particularly useful for the treatment of PNH in a limited number of patients with mutations in the C5 gene that prevent eculizumab from binding to C5. An example of such a patient is a single missense C5 heterozygous mutation c.2654G->A, which predicts the polymorphism p.Arg885His (for a description of this polymorphism, see Nishimura, J. et al., The New England Journal of Medicine, 2014, Vol. 370, No. 7, pp. 632-639, the entire content of which is incorporated herein by reference). Similar to eculizumab, R5000 blocks the proteolytic cleavage of C5 to C5a and C5b. Unlike eculizumab, R5000 can also bind to C5b and block its association with C6, thereby preventing subsequent MAC assembly. Therefore, advantageously, any C5b resulting from incomplete inhibition by R5000 is prevented from binding to C6 and completing the assembly of the MAC.

[0068] In some cases, R5000 can be used as a therapeutic alternative to eculizumab in patients with PNH, potentially providing greater efficacy without the inconvenience and disadvantages of IV administration, as well as the known risks of immunogenicity and hypersensitivity associated with monoclonal antibodies. Furthermore, serious complications of long-term IV administration, such as infections, decreased venous access, local thrombosis, and hematomas, can be overcome with R5000 administered by subcutaneous (SC) injection.

[0069] Inflammatory indications In some embodiments, the compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat subjects having inflammation-related diseases, disorders, and / or pathological conditions. Inflammation can be upregulated during the complement system proteolytic cascade. While inflammation can have beneficial effects, excessive inflammation can lead to various pathological conditions (Markiewski, 2007, American Journal of Pathology, Vol. 17, pp. 715-727). Therefore, the compounds and compositions according to the present invention may be used to reduce or eliminate inflammation associated with complement activation.

[0070] sterile inflammation In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat, prevent, or delay the onset of sterile inflammation. Sterile inflammation is inflammation that occurs in response to stimuli other than infection. Sterile inflammation can be a normal response to stress such as genomic stress, hypoxic stress, nutrient stress, and endoplasmic reticulum stress, which are caused by physical, chemical, or metabolic noxious stimuli. Sterile inflammation can contribute to the pathogenesis of many diseases, such as, but not limited to, ischemia-induced injury, rheumatoid arthritis, acute lung injury, drug-induced liver injury, inflammatory bowel disease, and / or other diseases, disorders, or pathologies. Methods and compositions for the treatment, prevention, and / or delay of the mechanisms and symptoms of aseptic inflammation may include those taught in Rubartelli, Frontiers in Immunology, 2013, Vol. 4, pp. 398-99; Rock et al., Annual Review of Immunology, 2010, Vol. 28, pp. 321-342; or U.S. Patent No. 8,101,586 (the contents of which are incorporated herein by reference in whole).

[0071] Systemic inflammatory response (SIRS) and sepsis In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used for the treatment and / or prevention of systemic inflammatory response syndrome (SIRS). SIRS is inflammation affecting the entire body. When SIRS is caused by infection, it is referred to as sepsis. SIRS can also be caused by non-infectious events such as trauma, injury, burns, ischemia, bleeding, and / or other pathological conditions. Among the negative outcomes associated with SIRS and / or sepsis is multiple organ failure (MOF). Complement inhibition at the C3 level in Gram-negative sepsis not only significantly protects organs against Escherichia coli (E. coli)-induced progressive MOF but also interferes with bacterial clearance. The compounds and compositions described herein include C5 complement component inhibitors that may be administered to subjects with sepsis to provide the benefits of organ protection without adversely altering bacterial clearance.

[0072] In some embodiments, this disclosure provides a method for treating sepsis. Sepsis may be induced by a microbial infection. The microbial infection may include at least one Gram-negative infectious pathogen. As used herein, the term “infectious pathogen” means any entity that invades or otherwise infects cells, tissues, organs, compartments, or bodily fluids of a sample or subject. In some cases, the infectious pathogen may be a bacterium, a virus, or another pathogen. A Gram-negative infectious pathogen is a Gram-negative bacterium. A Gram-negative infectious pathogen may include, but is not limited to, Escherichia coli (E. coli).

[0073] A method for treating sepsis may involve administering one or more C5 inhibitors to a subject. The C5 inhibitor may be R5000. According to some methods, complement activation may be reduced or inhibited. The reduction or inhibition of complement activation may be determined by detecting one or more products of complement activity in the subject sample. Such products may include C5 cleavage products (e.g., C5a and C5b) or downstream complexes (e.g., C5b-9) formed as a result of C5 cleavage. In some embodiments, the disclosure provides a method for treating sepsis using R5000, where the levels of C5a and / or C5b-9 are reduced or removed in and / or in at least one sample obtained from the subject. For example, C5a and / or C5b-9 levels may be reduced by approximately 0% to approximately 0.05%, approximately 0.01% to approximately 1%, approximately 0.05% to approximately 2%, approximately 0.1% to approximately 5%, approximately 0.5% to approximately 10%, approximately 1% to approximately 15%, approximately 5% to approximately 25%, approximately 10% to approximately 50%, approximately 20% to approximately 60%, approximately 25% to approximately 75%, and approximately 50% to approximately 100% when comparing subjects (or samples obtained from such subjects) that are not treated with R5000 (or subjects) (including subjects treated with other complement inhibitors), or when comparing with the same subjects (or samples) during the prior treatment period or the initial period of treatment.

[0074] In some embodiments, the C5b-9 levels reduced by R5000 treatment are C5b-9 levels associated with one or more of the classical pathway of complement activation, the alternative pathway of complement activation, and the lectin pathway of complement activation.

[0075] In some embodiments, the presence, absence, and / or levels of one or more factors associated with sepsis may be modulated by administering R5000 to subjects with sepsis. The presence or absence of such factors may be determined using assays for their detection. Changes in factor levels may be determined by measuring the levels of such factors in subjects with sepsis after R5000 treatment and comparing those levels to earlier levels in the same subjects (either before R5000 treatment or during one or more earlier periods of treatment) or in subjects not treated with R5000 (including subjects with sepsis who have not received any treatment or subjects receiving some other form of treatment). The comparison may be indicated by the percentage difference in factor levels between subjects treated with R5000 and those not treated with R5000.

[0076] C5 cleavage products may contain any proteins or complexes that may result from C5 cleavage. In some cases, C5 cleavage products may contain, but are not limited to, C5a and C5b. C5b cleavage products may proceed to the formation of complexes with complement proteins C6, C7, C8, and C9 (referred herein to as "C5b-9"). Therefore, C5 cleavage products containing C5b-9 may be detected and / or quantified to determine whether complement activity is reduced or inhibited. Detection of C5b-9 precipitation may be carried out, for example, by using the Wieslab® ELISA (Euro Diagnostica, Malmö, Sweden) kit. For the quantification of cleavage products, measurement may be performed using "complement arbitrary units" (CAU), as described by others (see, for example, Bergseth G. et al., 2013, Molecular Immunology (Mol Immunol.), Vol. 56, pp. 232-239 (the contents of which are incorporated herein by reference in their entirety)).

[0077] In some embodiments, C5b-9 production may be reduced or inhibited by treating sepsis with a C5 inhibitor (e.g., R5000). According to the present invention, administration of R5000 to a subject may result in modification of bacterial clearance in the subject and / or in at least one sample obtained from the subject. Bacterial clearance, as referred herein, is the partial or complete removal / reduction of bacteria from a subject or sample. Clearance may occur through killing bacteria or otherwise rendering them unable to grow and / or reproduce. In some cases, bacterial clearance may occur through lysis and / or immunodestruction (e.g., through phagocytosis, bacterial cytolysis, opsonization, etc.). According to some methods, bacterial clearance in a subject treated with a C5 inhibitor (e.g., R5000) may have no effect on bacterial clearance or a beneficial effect. This may result from the absence or reduction of the effect on C3b levels by C5 inhibition. In some embodiments, a method of treating sepsis with R5000 may avoid interference with or enhance C3b-dependent opsonization.

[0078] In some cases, bacterial clearance associated with R5000 treatment may be enhanced compared to bacterial clearance in untreated subjects or subjects treated with other forms of complement inhibitors, such as C3 inhibitors. In some embodiments, subjects with sepsis treated with R5000 may experience 0% to at least 100% enhanced bacterial clearance when compared to bacterial clearance in untreated subjects (including subjects treated with other complement inhibitors), or when compared to earlier bacterial clearance levels in the same subjects before or during earlier treatment periods with R5000. For example, bacterial clearance in subjects treated with R5000 and / or in at least one sample obtained from such subjects may be increased by approximately 0% to approximately 0.05%, approximately 0.01% to approximately 1%, approximately 0.05% to approximately 2%, approximately 0.1% to approximately 5%, approximately 0.5% to approximately 10%, approximately 1% to approximately 15%, approximately 5% to approximately 25%, approximately 10% to approximately 50%, approximately 20% to approximately 60%, approximately 25% to approximately 75%, and approximately 50% to approximately 100% when compared to subjects not treated with R5000 (including subjects treated with other complement inhibitors) and / or when compared to samples obtained from such subjects, or when compared to the same subjects during the pre-treatment period or the early period of treatment and / or when compared to samples obtained from the same subjects during the pre-treatment period or the early period of treatment.

[0079] Bacterial clearance may be measured in a subject by directly measuring bacterial levels in and / or in a sample of the subject, or by measuring one or more indicators of bacterial clearance (e.g., the level of bacterial components released after lysis). The bacterial clearance level may then be determined by comparison with previous measurements of bacterial / indicator levels or with bacterial / indicator levels in subjects that are untreated or receiving different treatments. In some cases, colony-forming units (CFUs) from collected blood (e.g., to obtain blood at CFU / ml) are tested to determine bacterial levels.

[0080] In some embodiments, sepsis treatment using R5000 may be performed without effect on phagocytosis or without substantial impairment of phagocytosis. This may include neutrophil-dependent and / or monocyte-dependent phagocytosis. Unimpaired or substantially unimpaired phagocytosis due to R5000 treatment may be due to limited or absent changes in C3b levels associated with R5000 treatment.

[0081] Oxidative burst is a C5a-dependent process characterized by the production of peroxides by certain cells, particularly macrophages and neutrophils, after pathogen loading (see Mollnes et al., 2002, Blood, Vol. 100, pp. 1869-1877, the contents of which are incorporated herein by reference in their entirety).

[0082] In some embodiments, oxidative bursts may be reduced or prevented in subjects with sepsis after treatment with R5000. This may be due to a decrease in C5a levels due to R5000-dependent C5 inhibition. Oxidative bursts may be reduced by approximately 0% to approximately 0.05%, approximately 0.01% to approximately 1%, approximately 0.05% to approximately 2%, approximately 0.1% to approximately 5%, approximately 0.5% to approximately 10%, approximately 1% to approximately 15%, approximately 5% to approximately 25%, approximately 10% to approximately 50%, approximately 20% to approximately 60%, approximately 25% to approximately 75%, and approximately 50% to approximately 100% in subjects administered with R5000 compared with subjects not treated with R5000 (including subjects treated with other complement inhibitors), or compared with the same subjects during the prior treatment period or the early period of treatment.

[0083] Lipopolysaccharide (LPS) is a known immunostimulatory factor and a component of the bacterial cell coat. Complement-dependent lysis results in LPS release, which can contribute to inflammatory responses, such as those characteristic of sepsis. In some embodiments, treatment of sepsis with R5000 may reduce LPS levels. This may be due to a decrease in complement-mediated lysis caused by inhibition of C5-dependent complement activity. In some embodiments, LPS levels in subjects administered with R5000 (or in samples obtained from such subjects) may be reduced or eliminated by approximately 0% to approximately 0.05%, approximately 0.01% to approximately 1%, approximately 0.05% to approximately 2%, approximately 0.1% to approximately 5%, approximately 0.5% to approximately 10%, approximately 1% to approximately 15%, approximately 5% to approximately 25%, approximately 10% to approximately 50%, approximately 20% to approximately 60%, approximately 25% to approximately 75%, and approximately 50% to approximately 100% when compared to subjects (or samples obtained from such subjects) that are not treated with R5000 (or control samples) (including subjects treated with other complement inhibitors), or when compared to the same subjects (or control samples) during the pre-treatment period or the early period of treatment.

[0084] In some embodiments, LPS levels in subjects (or control samples) with sepsis treated with R5000 can be reduced by 100% compared to subjects (or control samples) with sepsis not treated with R5000 (including subjects receiving one or more other forms of treatment), or compared to the same subjects (or control samples) during the pre-treatment period or the early period of treatment.

[0085] In some embodiments of this disclosure, the sepsis-inducing levels of one or more cytokines may be reduced in conjunction with R5000 treatment. Cytokines include numerous cellular signaling molecules that stimulate an immune response to infection. A "cytokine storm" is a dramatic upregulation of at least four cytokines, interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor α (TNFα), resulting from bacterial infection and potentially contributing to sepsis. C5a is known to induce the synthesis and activity of these cytokines. Therefore, inhibitors of C5 may reduce cytokine levels by reducing C5a levels. Cytokine levels may be evaluated in subjects or subject samples to assess the ability of C5 inhibitors to reduce the levels of one or more inflammatory cytokines that are upregulated during sepsis. IL-6, IL-8, MCP-1, and / or TNFα levels may decrease by approximately 0% to 0.05%, 0.01% to 1%, 0.05% to 2%, 0.1% to 5%, 0.5% to 10%, 1% to 15%, 5% to 25%, 10% to 50%, 20% to 60%, 25% to 75%, and 50% to 100% in subjects treated with R5000 compared to subjects not treated with R5000 (including subjects treated with other complement inhibitors), or compared to the same subjects during the prior treatment period or the early stages of treatment. In some embodiments, IL-6, IL-8, MCP-1, and / or TNFα levels may be reduced by 100% in subjects with sepsis treated with R5000 compared to subjects with sepsis not treated with R5000 (including subjects receiving one or more other forms of treatment), or compared to the same subjects during the prior treatment period or the early period of treatment.

[0086] One complication associated with sepsis is dysregulation of the coagulation and / or fibrinolytic pathways (Levi M. et al., 2013, Seminars in thrombosis and hemostasis, Vol. 39, pp. 559-566; Rittirsch D. et al., 2008, Nature Reviews Immunology, Vol. 8, pp. 776-777; and Dempfle C., 2004, A Thromb Haemost., Vol. 91, No. 2, pp. 213-24; the contents of each of these are incorporated herein by reference in their entirety). While controlled local activation of these pathways is important for defense against pathogens, uncontrolled systemic activation can be harmful. Complement activity associated with bacterial infections can promote dysregulation of coagulation and / or fibrinolysis due to increased host cell and tissue damage associated with MAC formation. In some embodiments, treatment of sepsis with R5000 can normalize the coagulation and / or fibrinolysis pathways.

[0087] Dysregulation of coagulation and / or fibrinolysis associated with sepsis may include disseminated intravascular coagulation (DIC). DIC is a condition that results in tissue and organ damage due to activated coagulation and clot formation in small vessels. This activity reduces blood flow to tissues and organs and consumes blood factors necessary for coagulation in other parts of the body. The absence of these blood factors in the bloodstream can lead to uncontrolled bleeding in other parts of the body. In some embodiments, DIC may be reduced or eliminated by treatment of sepsis with R5000.

[0088] Coagulation disorders associated with sepsis may be detected by measuring activated partial thromboplastin time (APTT) and / or prothrombin time (PT). These are tests performed on plasma samples to determine whether coagulation factor levels are low. In subjects with DIC, APTT and / or PT are prolonged due to decreased levels of coagulation factors. In some embodiments, treatment of sepsis subjects with R5000 may reduce and / or normalize APTT and / or PT in samples obtained from the treated subjects.

[0089] Sepsis-associated coagulation disorders may be further evaluated through analysis of thrombin-antithrombin (TAT) complex levels and / or tissue factor (TF) mRNA leukocyte expression. Elevated TAT complex levels and TF mRNA leukocyte expression are associated with coagulation disorders and are consistent with DIC. In some embodiments, treatment of sepsis with R5000 may result in reductions of approximately 0.005% to 0.05%, 0.01% to 1%, 0.05% to 2%, 0.1% to 5%, 0.5% to 10%, 1% to 15%, 5% to 25%, 10% to 50%, 20% to 60%, 25% to 75%, and 50% to 100% in TAT levels and / or leukocyte TF mRNA levels when compared to subjects not treated with R5000 (including subjects treated with other complement inhibitors), or to the same subjects during the prior treatment period or the early stages of treatment. In some embodiments, TAT levels and / or leukocyte TF mRNA levels may be reduced by 100% in subjects with sepsis treated with R5000 compared to subjects with sepsis not treated with R5000 (including subjects receiving one or more other forms of treatment), or compared to the same subjects during the prior treatment period or the early period of treatment.

[0090] Factor XII is a crucial factor for normal blood coagulation in plasma. Factor XII levels may be reduced in plasma samples taken from subjects with coagulation disorders (e.g., DIC) due to the consumption of factor XII related to coagulation in small vessels. In some embodiments, sepsis treatment with R5000 may reduce factor XII consumption. Therefore, factor XII levels may be elevated in plasma samples taken from subjects with sepsis after R5000 treatment. Factor XII levels in plasma samples may increase by approximately 0.005% to 0.05%, 0.01% to 1%, 0.05% to 2%, 0.1% to 5%, 0.5% to 10%, 1% to 15%, 5% to 25%, 10% to 50%, 20% to 60%, 25% to 75%, and 50% to 100% when compared to subjects not treated with R5000 (including subjects treated with other complement inhibitors), or when compared to plasma samples taken from the same subjects during the prior treatment period or the early stages of treatment. In some embodiments, factor XII levels may be 100% elevated in plasma samples from subjects with sepsis treated with R5000 compared to plasma samples from subjects with sepsis not treated with R5000 (including subjects receiving one or more other forms of treatment), or compared to plasma samples taken from the same subjects during the pre-treatment period or the early stages of treatment.

[0091] Fibrinolysis is the breakdown of fibrin due to enzymatic activity, a critical process for blood clot formation. Dysregulation of fibrinolysis can occur in severe sepsis, and it has been reported to affect normal coagulation in baboons infected with Escherichia coli (P. de Boer JP et al., 1993, Circulatory Shock, Vol. 39, pp. 59-67, the entire content of which is incorporated herein by reference). Plasma indicators of sepsis-dependent fibrinolytic dysfunction (including, but not limited to, DIC-associated, fibrinolytic dysfunction) may include, but are not limited to, decreased fibrinogen levels (indicating a reduced ability to form fibrin clots), increased tissue plasminogen activator (tPA) levels, increased plasminogen activator inhibitor type 1 (PAI-1) levels, increased plasmin-anti-plasmin (PAP) levels, increased fibrinogen / fibrin degradation products, and increased D-dimer levels. In some embodiments, treatment of sepsis with R5000 may result in a decrease in plasma fibrinogen levels and / or an increase in plasma levels of tPA, PAI-1, PAP, fibrinogen / fibrin degradation products, and / or D-dimers when compared to levels in plasma samples from subjects not treated with R5000 (including subjects treated with other complement inhibitors), or when compared to levels in plasma samples taken from the same subjects during the prior treatment period or the early stages of treatment, by approximately 0.005% to approximately 0.05%, approximately 0.01% to approximately 1%, approximately 0.05% to approximately 2%, approximately 0.1% to approximately 5%, approximately 0.5% to approximately 10%, approximately 1% to approximately 15%, approximately 5% to approximately 25%, approximately 10% to approximately 50%, approximately 20% to approximately 60%, approximately 25% to approximately 75%, and approximately 50% to approximately 100%. In some embodiments, sepsis-related decreases in plasma fibrinogen levels and / or sepsis-related increases in plasma levels of tPA, PAI-1, PAP, fibrinogen / fibrin degradation products, and / or D-dimers may differ by at least 10,000% when compared to levels in plasma samples from subjects with sepsis treated with R5000.

[0092] Another consequence of excessive complement activity associated with sepsis is a decrease in red blood cells due to complement-dependent hemolysis and / or C3b-dependent opsonization. A method for treating sepsis using R5000 according to this disclosure may include reducing complement-dependent hemolysis. One method for evaluating complement-dependent hemolysis associated with sepsis includes obtaining a complete blood count. The complete blood count may be obtained through an automated process that counts the cell types present in a blood sample. Results from complete blood count analysis typically include hematocrit, red blood cell (RBC) count, white blood cell (WBC) count, and platelet levels. The hematocrit level is used to determine the percentage of blood (by volume) composed of red blood cells. Hematocrit levels, platelet levels, RBC levels, and WBC levels may be decreased in sepsis due to hemolysis. In some embodiments, treatment of sepsis with R5000 results in an increase in hematocrit levels, platelet levels, RBC levels, and / or WBC levels. This increase may be immediate or may occur over time with treatment (e.g., single or multiple doses).

[0093] In some embodiments, treatment of a subject with R5000 may reduce the activation of sepsis-related leukocytes (e.g., neutrophils and macrophages). When used herein in relation to leukocytes, "activation" refers to the recruitment and / or maturation of these cells to perform related immune functions. The reduction in leukocyte activation by R5000 treatment may be determined by evaluating the subject being treated or a sample obtained from the subject being treated.

[0094] In some embodiments, treatment of sepsis with R5000 may improve one or more vital signs in the patient during treatment. Such vital signs may include, but are not limited to, heart rate, mean systemic arterial pressure (MSAP), respiratory rate, oxygen saturation, and body temperature.

[0095] In some embodiments, treatment of sepsis with R5000 may stabilize or reduce sepsis-related capillary leakage and / or endothelial barrier dysfunction (i.e., maintain or improve capillary leakage and / or endothelial barrier dysfunction). Stabilization or reduction of capillary leakage and / or endothelial barrier dysfunction may be determined by measuring total plasma protein levels and / or plasma albumin levels. An increase in either level compared to sepsis-related plasma levels may indicate a decrease in capillary leakage. Therefore, treatment of sepsis with R5000 may increase total plasma protein and / or plasma albumin levels.

[0096] The methods of this disclosure may include methods for treating sepsis using R5000, in which the level of one or more acute-phase proteins is reduced. Acute-phase proteins are proteins produced by the liver under inflammatory conditions. R5000 treatment may reduce sepsis-related inflammation and result in reduced hepatic production of acute-phase proteins.

[0097] According to some methods of the present invention, treatment with R5000 may reduce, reverse, or prevent sepsis-induced organ injury and / or organ dysfunction. Indicators that may be reduced along with improved organ function may include, but are not limited to, plasma lactate (indicating improved intravascular perfusion and clearance), creatinine, blood urea nitrogen (both indicating improved renal function), and hepatic transaminase (indicating improved hepatic function). In some embodiments, in subjects treated for sepsis with R5000, the risk of fever, secondary infection, and / or sepsis recurrence is reduced.

[0098] The methods of this disclosure may include preventing sepsis-related death and / or improving the survival time of subjects suffering from sepsis through treatment with R5000. The improved survival time may be determined by comparing the survival time of subjects treated with R5000 with the survival time of untreated subjects (including subjects treated with one or more other forms of treatment). In some embodiments, the survival time is extended by at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 1 month, at least 2 months, at least 4 months, at least 6 months, at least 1 year, at least 2 years, at least 5 years, or at least 10 years.

[0099] In some embodiments, R5000 is administered as a single dose. In some embodiments, R5000 is administered as multiple doses. For example, R5000 may consist of an initial dose followed by one or more repeat doses. Repeat doses may be administered approximately 1 to 24 hours, 2 to 48 hours, 4 to 72 hours, 8 to 96 hours, 12 to 36 hours, or 18 to 60 hours after a previous dose. In some cases, repeat doses may be administered 1, 2, 3, 4, 5, 6, 7 days, 2 weeks, 4 weeks, 2 months, 4 months, 6 months, or more than 6 months after a previous dose. In some cases, repeat doses may be administered in response to a need to stabilize or reduce sepsis in the subject or to stabilize or reduce one or more sepsis-related effects. The repeated dose may contain the same amount of R5000 or a different amount.

[0100] The compounds and compositions according to the present invention may be used for the control and / or balancing of complement activation for the prevention and treatment of SIRS, sepsis, and / or MOF. Methods of applying complement inhibitors for the treatment of SIRS and sepsis may include those described in U.S. Patent Application Publication No. 2013 / 0053302 or U.S. Patent No. 8,329,169 (each of which is incorporated herein by reference in whole).

[0101] Acute respiratory distress syndrome (ARDS) In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used for the treatment and / or prevention of the onset of acute respiratory distress syndrome (ARDS). ARDS is a widespread inflammation of the lungs that can be caused by trauma, infection (e.g., sepsis), severe pneumonia, and / or inhalation of harmful substances. ARDS is typically a serious, life-threatening complication. Studies suggest that neutrophils may contribute to the development of ARDS by influencing the accumulation of polymorphonuclear cells in the damaged alveolar and interstitial tissues of the lungs. Therefore, compounds and compositions according to the present invention may be administered to reduce and / or prevent tissue factor production in alveolar neutrophils. Compounds and compositions according to the present invention may, in some cases, be used for the treatment, prevention, and / or delay of ARDS in accordance with any of the methods taught in International Publication No. 2009 / 014633 (the contents of which are incorporated herein by reference in their entirety).

[0102] Periodontitis In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat or prevent the development of periodontitis and / or related conditions. Periodontitis is a generalized chronic inflammation that results in the destruction of periodontal tissues, which are the tissues that surround and support the teeth. The condition also includes alveolar bone loss (the bone that holds the teeth together). Periodontitis can be caused by a lack of oral hygiene, which leads to the accumulation of bacteria at the gingival margin, also known as plaque. Certain health conditions, such as diabetes or malnutrition and / or habits such as smoking, can increase the risk of periodontitis. Periodontitis can increase the risk of seizures, myocardial infarction, atherosclerosis, diabetes, osteoporosis, premature birth, and even other health problems. Studies have demonstrated a correlation between periodontitis and local complement activity. Periodontal bacteria can inhibit or activate certain components of the complement cascade. Therefore, compounds and compositions according to the present invention may be used to prevent and / or treat periodontitis and related diseases and conditions. Complement activation inhibitors and therapeutic methods may include any of those taught by Hajishengallis, Biochem Pharmacol. 2010, 15;80(12):1 and Lambris or U.S. Patent Application Publication No. 2013 / 0344082 (each of which is incorporated herein by reference in whole).

[0103] dermatomyositis In some embodiments, the compounds, compositions, and / or methods of the present invention, such as pharmaceutical compositions, may be used to treat dermatomyositis. Dermatomyositis is an inflammatory muscle disease characterized by muscle weakness and chronic myositis. Dermatomyositis often begins with a concurrently associated skin rash or precedes muscle weakness. The compounds, compositions, and / or methods of the present invention may be used to reduce or prevent dermatomyositis.

[0104] Wounds and injuries Compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat and / or promote the healing of various types of wounds and / or injuries. As used herein, the term “injury” typically means physical trauma, but may include localized infection or disease processes. Injuries may be characterized by damage, injury, or destruction caused by external events affecting parts of a living organism and / or organs. Wounds relate to cuts, bruises, burns, and / or other impacts to the skin that result in destruction or damage to the skin. Wounds and injuries are often acute, but if not properly healed, they can lead to chronic complications and / or inflammation.

[0105] Wounds and burn wounds In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used for the treatment and / or promotion of wound healing. Healthy skin provides a waterproof protective barrier against pathogens and other environmental agents. Skin also controls body temperature and fluid evaporation. When skin is damaged, these functions are disrupted, making skin healing difficult. Damage triggers a group of physiological processes related to the immune system that restore and regenerate tissue. Complement activation is one of these processes. As taught by van de Goot et al., Journal of Burn Care Res, 2009, Vol. 30, pp. 274-280, and Cazander et al., Clinical and Developmental Immunology, 2012, Vol. 2012, pp. 534-291 (the contents of which are incorporated herein by reference in their entirety), studies of complement activation have identified several complement components involved in wound healing. In some cases, excessive complement activation can lead to cell death and increased inflammation (potentially resulting in impaired wound healing and chronic wounds). In some cases, the compounds and compositions according to the present invention can be used to reduce or eliminate such complement activation and promote wound healing. Treatment with the compounds and compositions according to the present invention may be carried out in accordance with any of the methods for treating wounds disclosed in International Publication No. 2012 / 174055 (the contents of which are incorporated herein by reference in their entirety).

[0106] head injury In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat and / or promote the healing of head injuries. Head injuries include injuries to the scalp, skull, or brain. Examples of head injuries include, but are not limited to, concussions, contusions, skull fractures, traumatic brain injuries, and / or other injuries. Head injuries can be mild or severe. In some cases, head injuries may result in long-term physical and / or psychological complications or death. Studies suggest that head injuries may induce inappropriate intracranial complement cascade activation, which may lead to a local inflammatory response and cause secondary brain damage resulting in cerebral edema and / or neuronal death (Stahel et al., Brain Research Reviews, 1998, Vol. 27, pp. 243-2456 (the entire content of which is incorporated herein by reference)). The compounds and compositions according to the present invention may be used for the treatment of head trauma and / or to reduce or prevent associated secondary complications. Methods of using the compounds and compositions according to the present invention to control complement cascade activation in head trauma may include any of those taught by Holers et al. in U.S. Patent No. 8,911,733 (the contents of which are incorporated herein by reference in their entirety).

[0107] crush injury In some embodiments, the compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat and / or promote the healing of crush injuries. Crush injuries are injuries caused by force or pressure applied to the body, resulting in bleeding, bruises, fractures, nerve damage, wounds, and / or other bodily injuries. The compounds and compositions according to the present invention may be used to promote post-crush injury healing (e.g., promoting nerve regeneration, promoting fracture healing, preventing or treating inflammation and / or other related complications) by reducing complement activation after a crush injury. The compounds and compositions according to the present invention may be used to promote treatment in accordance with any of the methods taught in U.S. Patent No. 8,703,136, International Publication No. 2012 / 162215, International Publication No. 2012 / 174055, or U.S. Patent Application Publication No. 2006 / 0270590 (each of which is incorporated herein by reference in whole).

[0108] Ischemia / reperfusion injury In some embodiments, the compounds, compositions, and / or methods of the Disclosure may be used to treat injuries associated with ischemia and / or reperfusion, which may be accompanied by surgical intervention (e.g., transplantation). Accordingly, the compounds, compositions, and / or methods of the Disclosure may be used to reduce or prevent ischemic and / or reperfusion injury.

[0109] autoimmune diseases Compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat subjects with autoimmune diseases and / or disorders. The immune system can be divided into innate and adaptive systems, representing nonspecific immediate defense mechanisms and more complex antigen-specific systems, respectively. The complement system is part of the innate immune system and recognizes and eliminates pathogens. In addition, complement proteins can modulate adaptive immunity, linking innate and adaptive responses. Autoimmune diseases and disorders are immune abnormalities in which the system targets the body's own tissues and substances. Autoimmune diseases may involve certain tissues or organs in the body. Compounds and compositions according to the present invention may be used to modulate complement in the treatment and / or prevention of autoimmune diseases. In some cases, such compounds and compositions may be used according to the methods presented in Ballanti et al., Immunologic Research, 2013, Vol. 56, pp. 477-491 (the contents of which are incorporated herein by reference in their entirety).

[0110] Antiphospholipid syndrome (APS) and fulminant antiphospholipid syndrome (CAPS) In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat antiphospholipid syndrome (APS) by controlling complement activation. APS is an autoimmune condition caused by antiphospholipid antibodies that cause blood clotting. APS can cause recurrent venous or arterial thrombosis in organs and complications in the placental circulation, potentially leading to pregnancy-related complications such as miscarriage, stillbirth, pre-eclampsia, preterm birth, and / or other complications. Fulminant antiphospholipid syndrome (CAPS) is an extreme acute form of a similar condition that simultaneously causes venous occlusion in several organs. Studies suggest that complement activation may contribute to APS-related complications, including pregnancy-related complications, thrombotic (coagulation) complications, and vascular complications. Compounds and compositions according to the present invention may be used to treat APS-related conditions by reducing or eliminating complement activation. In some cases, the compounds and compositions according to the present invention may be used to treat APS and / or APS-related complications in accordance with the methods taught by Salmon et al., Annals of the Rheumatic Disease, 2002, Vol. 61 (Suppl II), ii46-ii50, and Mackworth-Young, Clinical and Experimental Immunology, 2004, Vol. 136, pp. 393-401.

[0111] cold agglutinin disease In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat cold agglutinin disease (CAD), also referred to as cold agglutinin-mediated hemolysis. CAD is an autoimmune disease caused by high concentrations of IgM antibodies that interact with red blood cells at low body temperatures [Engelhardt et al., Blood, 2002, Vol. 100, No. 5, pp. 1922-23]. CAD can lead to conditions such as anemia, fatigue, dyspnea, hemoglobinuria, and / or acrocyanosis. CAD is associated with robust complement activation, and studies have shown that CAD can be treated with complement inhibitor therapy. Therefore, the present invention provides a method for treating CAD using compounds and compositions according to the present invention. In some cases, the compounds and compositions according to the present invention may be used to treat CAD in accordance with the methods taught in Roth et al., Blood, 2009, Vol. 113, pp. 3885-86, or in International Publication No. 2012 / 139081 (each of which is incorporated herein by reference in whole).

[0112] myasthenia gravis In some embodiments, the compounds, compositions, and / or methods of the present invention may be used to treat myasthenia gravis, a neuromuscular disease caused by autoimmunity. The compounds, compositions, and / or methods of the present invention may be used to reduce or prevent neuromuscular problems associated with myasthenia gravis.

[0113] Guillain-Barré syndrome In some embodiments, the compounds, compositions, and methods of the present invention, such as pharmaceutical compositions, may be used to treat Guillain-Barré syndrome (GBS), an autoimmune disease involving autoimmune attacks of the peripheral nervous system. The compounds, compositions, and / or methods of the present invention may be used to reduce or prevent peripheral nerve problems associated with GBS.

[0114] Vascular indications In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat vascular indications affecting blood vessels (e.g., arteries, veins, and capillaries). Such indications may affect blood circulation (blood pressure), blood flow, organ function, and / or other bodily functions.

[0115] Thrombotic microangiopathy (TMA) In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat and / or prevent thrombotic microangiopathy (TMA) and related diseases. Microangiopathy affects the body's microvessels (capillaries), thickening and weakening the capillary walls, making them more susceptible to bleeding and slow blood circulation. TMA tends to lead to the development of vascular thrombosis, endothelial cell damage, thrombocytopenia, and hemolysis. Organs such as the brain, kidneys, muscles, gastrointestinal system, skin, and lungs may be affected. TMA may result from medical procedures and / or conditions, including, but not limited to, hematopoietic stem cell transplantation (HSCT), renal impairment, diabetes, and / or other conditions. TMA can be caused by underlying complement system dysfunction, as described in Meri et al., European Journal of Internal Medicine, 2013, Vol. 24, pp. 496-502 (the contents of which are incorporated herein by reference in their entirety). Generally, TMA can result from elevated levels of certain complement components that lead to thrombosis. In some cases, this can be caused by mutations in complement proteins or related enzymes. The resulting complement dysfunction can lead to complement targeting of endothelial cells and platelets, which can also lead to an increased risk of thrombosis. In some embodiments, TMA can be prevented and / or treated using compounds and compositions according to the present invention. In some cases, a person treating TMA with the compounds and compositions according to the present invention may do so in accordance with the description in U.S. Patent Publication No. 2012 / 0225056 or U.S. Patent Publication No. 2013 / 0246083, the contents of which are incorporated herein by reference in their entirety.

[0116] Disseminated intravascular coagulation (DIC) In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to treat and / or prevent disseminated intravascular coagulation (DIC) by controlling complement activation. DIC is a pathological condition in which the coagulation cascade is extensively activated in the blood, leading to the formation of blood clots, particularly in the capillaries. DIC can cause obstruction of blood flow to tissues, ultimately damaging organs. In addition, DIC can affect the normal process of blood coagulation, leading to serious bleeding. Compounds and compositions according to the present invention may be used to treat, prevent, or reduce the severity of DIC by modulating complement activity. In some cases, compounds and compositions according to the present invention may be used in accordance with any of the methods for treating DIC taught in U.S. Patent No. 8,652,477 (the contents of which are incorporated herein by reference in their entirety).

[0117] vasculitis In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat vasculitis. Generally, vasculitis is a disorder associated with inflammation of blood vessels, including veins and arteries, characterized by leukocytes attacking tissues and causing dilation of blood vessels. Vasculitis can be associated with infection, as in the case of Rocky Mountain spotted fever or autoimmune diseases. An example of autoimmune-associated vasculitis is anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis. ANCA vasculitis is caused by abnormal antibodies that attack the body's own cells and tissues. ANCA attacks the cytoplasm of certain leukocytes and neutrophils, causing an attack on the blood vessel walls in certain organs and tissues of the body. ANCA vasculitis can affect the skin, lungs, eyes, and / or kidneys. Studies have shown that ANCA diseases activate complement accessory pathways, generating certain complement components that form an inflammatory amplification loop leading to vascular damage (Jennette et al., 2013, Seminars in Nephrology, Vol. 33, No. 6, pp. 557-564 (the entire content of which is incorporated herein by reference)). In some cases, the compounds and compositions according to the present invention may be used to prevent and / or treat ANCA vasculitis by inhibiting complement activation.

[0118] Atypical hemolytic uremic syndrome In some embodiments, the compounds, compositions, e.g., pharmaceutical compositions, and / or methods of the present disclosure may be useful for the treatment of atypical hemolytic uremic syndrome (aHUS), a rare disease caused by uncontrolled complement activation characterized by blood clot formation in small vessels. The compositions and methods of the present invention may be useful for reducing or inhibiting complement activation associated with aHUS.

[0119] Neurological indications The compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used for the prevention, treatment, and / or alleviation of symptoms of neurological indications, including neurodegenerative diseases and related disorders. Neurodegeneration generally involves loss of neuronal structure or function, including neuronal death. Such disorders may be treated by inhibiting the action of complement on neuronal cells using the compounds and compositions according to the present invention. Neurodegenerative disorders include, but are not limited to, amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), Parkinson's disease, and Alzheimer's disease.

[0120] Amyotrophic lateral sclerosis (ALS) In some embodiments, the compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used for the prevention, treatment, and / or symptom relief of ALS. ALS is a fatal motor neuron disease characterized by degeneration of spinal cord neurons, brainstem, and motor cortex. ALS causes muscle weakness that ultimately leads to respiratory failure. Since complement dysfunction may contribute to ALS, therapies using the compounds and compositions according to the present invention that target complement activity may prevent, treat, and reduce the symptoms of ALS. In some cases, the compounds and compositions according to the present invention may be used to promote nerve regeneration. In some cases, the compounds and compositions according to the present invention may be used as complement inhibitors in accordance with either of the methods taught in U.S. Patent Application Publication No. 2014 / 0234275 or U.S. Patent Application Publication No. 2010 / 0143344 (each of which is incorporated herein by reference in whole).

[0121] Alzheimer's disease In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat Alzheimer's disease by controlling complement activity. Alzheimer's disease is a chronic neurodegenerative disease with symptoms that may include disorientation, memory loss, mood swings, behavioral problems, and ultimately, loss of physical function. Alzheimer's disease is thought to be caused by extracellular brain accumulations of amyloid associated with inflammation-related proteins such as complement proteins (Sjoberg et al., 2009, Trends in Immunology, Vol. 30, No. 2, pp. 83-90 (the entire content of which is incorporated herein by reference)). In some cases, compounds and compositions according to the present invention may be used as complement inhibitors in accordance with any of the Alzheimer's disease therapies taught in U.S. Patent Application Publication No. 2014 / 0234275 (the entire content of which is incorporated herein by reference).

[0122] Kidney-related indications The compounds and compositions according to the present invention, such as pharmaceutical compositions, may in some cases be used to treat certain kidney-related diseases, disorders, and / or conditions by inhibiting complement activity. The kidneys are organs involved in removing metabolic waste products from the bloodstream. They are essential for various bodily functions as they control blood pressure, the urinary system, and homeostatic mechanisms. The kidneys may be more severely affected by inflammation due to their unique structural features and exposure to blood (compared to other organs). The kidneys also produce their own complement proteins, which can be activated by infection, kidney disease, and kidney transplantation. In some cases, the compounds and compositions according to the present invention may be used as complement inhibitors in the treatment of certain kidney diseases, conditions, and / or disorders, according to the methods taught in Quigg, Journal of Immunology (J Immunol), 2003, No. 171, pp. 3319-3324 (the contents of which are incorporated herein by reference in their entirety).

[0123] Lupus nephritis In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat lupus nephritis by inhibiting complement activity. Lupus nephritis is inflammation of the kidneys caused by an autoimmune disease called systemic lupus erythematosus (SLE). Symptoms of lupus nephritis include high blood pressure, foamy urine, swelling of the legs, feet, hands, or face, joint pain, muscle pain, fever, and rash. Lupus nephritis may be treated with inhibitors that control complement activity, including compounds and compositions according to the present invention. Methods and compositions for preventing and / or treating lupus nephritis by complement inhibition may include either of those taught in U.S. Patent Application Publication No. 2013 / 0345257 or U.S. Patent No. 8,377,437 (each of which is incorporated herein by reference in whole).

[0124] Membranoglomerulonephritis (MGN) In some embodiments, the compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat membranous glomerulonephritis (MGN) disorder by inhibiting the activation of certain complement components. MGN is a kidney disorder that can result in inflammation and structural changes. MGN is caused by antibodies that bind to soluble antigens in the renal capillaries (glomeruli). MGN may affect renal function, such as fluid filtration, and can lead to renal failure. The compounds and compositions according to the present invention may be used in accordance with the methods for preventing and / or treating MGN by complement inhibition taught in U.S. Patent Application Publication No. 2010 / 0015139 or International Publication No. 2000 / 021559 (each of which is incorporated herein by reference in its entirety).

[0125] hemodialysis complications In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat complications associated with hemodialysis by inhibiting complement activation. Hemodialysis is a medical procedure used to maintain kidney function in patients with renal failure. In hemodialysis, waste products derived from the blood, such as creatinine, urea, and free water, are removed externally. A common complication of hemodialysis treatment is chronic inflammation caused by contact between the blood and the dialysis membrane. Another common complication is thrombosis, which represents the formation of blood clots that obstruct blood circulation. Studies have suggested that these complications are related to complement activation. Hemodialysis may be combined with complement inhibitor therapy to provide a means of controlling inflammatory responses and pathological conditions, and / or preventing or treating thrombosis, in patients undergoing hemodialysis due to renal failure. Methods of using the compounds and compositions according to the present invention for the treatment of hemodialysis complications may be carried out in accordance with either the method taught by DeAngelis et al., Immunobiology, 2012, Vol. 217, No. 11, pp. 1097-1105, or by Kourtzelis et al., Blood, 2010, Vol. 116, No. 4, pp. 631-639 (the contents of which are incorporated herein by reference in their entirety).

[0126] eye disease In some embodiments, the compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat certain eye-related diseases, disorders, and / or conditions. In healthy eyes, the complement system is activated at low levels and continuously regulated by membrane-bound and soluble intraocular proteins that protect against pathogens. Complement activation therefore plays a crucial role in several eye-related complications, and control of complement activation may be used to treat such diseases. The compounds and compositions according to the present invention may be used as complement inhibitors in the treatment of eye diseases in accordance with any of the methods taught in Jha et al., Molecular Immunology (Mol Immunol.), 2007, Vol. 44, No. 16, pp. 3901-3908 or U.S. Patent No. 8,753,625 (each of which is incorporated herein by reference in whole).

[0127] Age-related macular degeneration (AMD) In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat age-related macular degeneration (AMD) by inhibiting intraocular complement activation. AMD is a chronic eye disease that causes central vision impairment, central vision blind spot, and / or ultimate central vision loss. Central vision affects the ability to read, drive, and / or recognize faces. AMD is generally classified into two types: non-exudative (dry) and exudative (wet). Dry AMD refers to deterioration of the macula, the central tissue of the retina. Wet AMD refers to subretinal vascular insufficiency that results in leakage of blood and fluids. (Jha et al., Molecular Immunology) As discussed in Immunol., 2007, Vol. 44, No. 16, pp. 3901-3908, several human and animal studies have identified complement proteins associated with AMD, and novel therapeutic strategies have involved the regulation of complement activation pathways. Methods according to the present invention, including the use of compounds and compositions according to the present invention for the prevention and / or treatment of AMD, may include either of the teachings in U.S. Patent Application Publication No. 2011 / 0269807 or U.S. Patent Application Publication No. 2008 / 0269318 (each of which is incorporated herein by reference in whole).

[0128] Corneal diseases In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat corneal diseases by inhibiting intraocular complement activation. The complement system plays a crucial role in protecting the cornea from pathogenic particles and / or inflammatory antigens. As the outermost front part of the eye, covering and protecting the iris, pupil, and anterior chamber, the cornea is exposed to external factors. Corneal diseases include, but are not limited to, keratoconus, keratitis, ocular herpes, and / or other diseases. Corneal complications can cause pain, blurred vision, tearing, redness, photophobia, and / or corneal scarring. While the complement system is crucial for corneal protection, complement activation, which involves the high expression of certain complement compounds, can cause damage to corneal tissue after the infection has been cleared. The present invention for modulating complement activity in the treatment of corneal diseases may include any of the methods taught in Jha et al., Molecular Immunology, 2007, Vol. 44, No. 16, pp. 3901-398.

[0129] Autoimmune uveitis In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat uveitis, an inflammation of the uveal layer of the eye. The uvea is the pigmented area of ​​the eye, including the choroid, iris, and ciliary body. Uveitis can cause redness, blurred vision, pain, adhesions, and ultimately blindness. Complement-activated products are present in the eyes of patients with autoimmune uveitis, and studies suggest that complement plays a crucial role in disease development. In some cases, compounds and compositions according to the present invention may be used to prevent and / or treat uveitis according to any of the methods identified in Jha et al., Molecular Immunology (Mol Immunol.), 2007, Vol. 44, No. 16, pp. 3901-3908 (the contents of which are incorporated herein by reference in their entirety).

[0130] diabetic retinopathy In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat diabetic retinopathy, a disease caused by changes in retinal blood vessels in diabetic patients. Retinopathy can cause vascular dilation and fluid leakage and / or abnormal vascular growth. Diabetic retinopathy can affect vision and ultimately lead to blindness. Studies have suggested that complement activation plays an important role in the progression of diabetic retinopathy. In some cases, compounds and compositions according to the present invention may be used in accordance with the treatment of diabetic retinopathy described by Jha et al., Molecular Immunology (Mol Immunol.), 2007, Vol. 44, No. 16, pp. 3901-398 (the contents thereof are incorporated herein by reference in their entirety).

[0131] Neuromyelitis optica (NMO) In some embodiments, the compounds, compositions, such as pharmaceutical compositions, and / or methods of the present invention may be used to treat neuromyelitis optica (NMO), an autoimmune disease that results in the destruction of the optic nerve. The compounds and / or methods of the present invention may also be used to prevent nerve destruction in subjects having NMO.

[0132] Sjögren's syndrome In some embodiments, the compounds, compositions, and / or methods of the present invention may be used to treat Sjögren's syndrome. Sjögren's syndrome is an eye disorder characterized by dry eyes that may cause flushing and / or itching. It is an autoimmune disorder in which the immune system targets glands in the eyes and mouth (which are involved in keeping those areas moist). The compounds, compositions, and / or methods of the present disclosure may be used to treat and / or reduce the symptoms of Sjögren's syndrome.

[0133] Pre-eclampsia and HELLP syndrome In some embodiments, compounds and compositions according to the present invention, such as pharmaceutical compositions, may be used to prevent and / or treat pre-eclampsia and / or HELLP syndrome (an abbreviation representing the syndrome features of 1) hemolysis, 2) elevated liver enzymes, and 3) low platelet count by complement inhibitor therapy. Pre-eclampsia is a pregnancy disorder characterized by symptoms including elevated blood pressure, bloating, shortness of breath, renal failure, hepatic dysfunction, and / or low platelet count. Pre-eclampsia is typically diagnosed by high urinary protein levels and hypertension. HELLP syndrome is a combination of hemolysis, elevated liver enzymes, and a low platelet condition. Hemolysis is a disorder involving the rupture of red blood cells, resulting in the release of hemoglobin from red blood cells. Elevated liver enzymes may represent pregnancy-induced hepatic conditions. Low platelet levels result in reduced coagulation ability and cause excessive bleeding. HELLP is associated with pre-eclampsia and liver dysfunction. HELLP syndrome typically occurs in later stages of pregnancy or after childbirth. It is typically diagnosed by blood tests that indicate the presence of three related conditions. HELLP is typically treated by inducing labor.

[0134] Studies suggest that complement activation occurs in HELLP syndrome and pre-eclampsia, and that complement components are present at elevated levels in HELLP and pre-eclampsia. Complement inhibitors can be used as therapeutic agents to prevent and / or treat these conditions. The compounds and compositions according to the present invention may be used in accordance with the methods for the prevention and / or treatment of HELLP and pre-eclampsia taught in Heager et al., Obstetrics & Gynecology, 1992, Vol. 79, No. 1, pp. 19-26 or in International Publication No. 201 / 078622 (the contents of which are incorporated herein by reference in their entirety).

[0135] formulation In some embodiments, the compounds or compositions of the present invention, such as pharmaceutical compositions, are formulated in an aqueous solution. In some cases, the aqueous solution further comprises one or more salts and / or one or more buffers. The salt may include sodium chloride, which may be present in concentrations of about 0.05 mM to about 50 mM, about 1 mM to about 100 mM, about 20 mM to about 200 mM, or about 50 mM to about 500 mM. Furthermore, the solution may contain at least 500 mM of sodium chloride. In some cases, the aqueous solution contains sodium phosphate. Sodium phosphate may be present in the aqueous solution in concentrations of about 0.005 mM to about 5 mM, about 0.01 mM to about 10 mM, about 0.1 mM to about 50 mM, about 1 mM to about 100 mM, about 5 mM to about 150 mM, or about 10 mM to about 250 mM. In some cases, a sodium phosphate concentration of at least 250 mM is used.

[0136] The compositions of the present invention may contain a C5 inhibitor at concentrations of approximately 0.001 mg / mL to approximately 0.2 mg / mL, approximately 0.01 mg / mL to approximately 2 mg / mL, approximately 0.1 mg / mL to approximately 10 mg / mL, approximately 0.5 mg / mL to approximately 5 mg / mL, approximately 1 mg / mL to approximately 20 mg / mL, approximately 15 mg / mL to approximately 40 mg / mL, approximately 25 mg / mL to approximately 75 mg / mL, approximately 50 mg / mL to approximately 200 mg / mL, or approximately 100 mg / mL to approximately 400 mg / mL. In some cases, the compositions of the present invention contain a C5 inhibitor at a concentration of at least 400 mg / mL.

[0137] The compositions of the present invention may contain a C5 inhibitor at any of the following concentrations, approximately, roughly, or precisely: 0.001 mg / mL, 0.2 mg / mL, 0.01 mg / mL, 2 mg / mL, 0.1 mg / mL, 10 mg / mL, 0.5 mg / mL, 5 mg / mL, 1 mg / mL, 20 mg / mL, 15 mg / mL, 40 mg / mL, 25 mg / mL, 75 mg / mL, 50 mg / mL, 200 mg / mL, 100 mg / mL, or 400 mg / mL. In some cases, the compositions of the present invention contain a C5 inhibitor at a concentration of at least 40 mg / mL.

[0138] In some embodiments, the compositions of the present invention include an aqueous composition comprising at least water and a C5 inhibitor (e.g., a cyclic C5 inhibitor polypeptide). The aqueous C5 inhibitor composition of the present invention may further comprise one or more salts and / or one or more buffers. In some cases, the aqueous composition of the present invention comprises water, a cyclic C5 inhibitor polypeptide, a salt, and a buffer.

[0139] The aqueous C5 inhibitor formulations of the present invention may have pH levels of approximately 2.0–3.0, approximately 2.5–3.5, approximately 3.0–4.0, approximately 3.5–4.5, approximately 4.0–5.0, approximately 4.5–5.5, approximately 5.0–6.0, approximately 5.5–6.5, approximately 6.0–7.0, approximately 6.5–7.5, approximately 7.0–8.0, approximately 7.5–8.5, approximately 8.0–9.0, approximately 8.5–9.5, or approximately 9.0–10.0. In some cases, the compounds and compositions of the present invention are prepared in accordance with Good Manufacturing Practices (GMP) and / or current GMP (cGMP). Guidelines used to implement GMP and / or cGMP may be obtained from one or more of the following sources: the U.S. Food and Drug Administration (FDA), the World Health Organization (WHO), and the International Conference on Harmonisation of Registration of Pharmaceuticals for Human Use (ICH).

[0140] Dosage and administration For the treatment of human subjects, C5 inhibitors may be formulated as pharmaceutical compositions. Depending on the subject to be treated, the method of administration, and the type of treatment desired (e.g., prevention, prophylaxis, or treatment), C5 inhibitors may be formulated in a manner that conforms to these parameters. Outlines of such techniques can be found in "Remington: The Science and Practice of Pharmacy," 21st edition, Lippincott, Williams & Wilkins, 2005; and "Encyclopedia of Pharmaceutical Technology," edited by J. Swarbrick and J.C. Boylan, 1988–1999, Marcel Dekker, New York, each of which is incorporated herein by reference.

[0141] The C5 inhibitors of the present invention may be provided in therapeutically effective doses. In some cases, therapeutically effective doses of the C5 inhibitors of the present invention can be achieved by administering one or more C5 inhibitors in doses of about 0.1 mg to about 1 mg, about 0.5 mg to about 5 mg, about 1 mg to about 20 mg, about 5 mg to about 50 mg, about 10 mg to about 100 mg, about 20 mg to about 200 mg, or at least 200 mg.

[0142] In some embodiments, a subject may be administered a therapeutic dose of a C5 inhibitor based on the subject's body weight. In some cases, the C5 inhibitor may be approximately 0.001 mg / kg to approximately 1.0 mg / kg, approximately 0.01 mg / kg to approximately 2.0 mg / kg, approximately 0.05 mg / kg to approximately 5.0 mg / kg, approximately 0.03 mg / kg to approximately 3.0 mg / kg, approximately 0.01 mg / kg to approximately 10 mg / kg, approximately 0.1 mg / kg to approximately 2.0 mg / kg, approximately 0.2 mg / kg to approximately 3.0 mg / kg, approximately 0.4 mg / kg to approximately 4.0 mg / kg, approximately 1.0 mg / kg to approximately 5.0 mg / kg, and approximately 2.0 mg The drug is administered in doses of approximately 4.0 mg / kg, 1.5 mg / kg to 7.5 mg / kg, 5.0 mg / kg to 15 mg / kg, 7.5 mg / kg to 12.5 mg / kg, 10 mg / kg to 20 mg / kg, 15 mg / kg to 30 mg / kg, 20 mg / kg to 40 mg / kg, 30 mg / kg to 60 mg / kg, 40 mg / kg to 80 mg / kg, 50 mg / kg to 100 mg / kg, or at least 100 mg / kg. This range may include a range suitable for administration to human subjects. The dose level may be highly dependent on the nature of the situation; drug efficacy; patient condition; practitioner's judgment; and frequency and method of administration.

[0143] In some cases, the C5 inhibitor of the present invention is provided at a concentration adjusted to achieve the desired level of the C5 inhibitor in a sample, a biological system, or a subject (e.g., plasma level in a subject). In some cases, the desired concentration of the C5 inhibitor in a sample, a biological system, or a subject may include a concentration of about 0.001 μM to about 0.01 μM, about 0.005 μM to about 0.05 μM, about 0.02 μM to about 0.2 μM, about 0.03 μM to about 0.3 μM, about 0.05 μM to about 0.5 μM, about 0.01 μM to about 2.0 μM, about 0.1 μM to about 50 μM, about 0.1 μM to about 10 μM, about 0.1 μM to about 5 μM, or about 0.2 μM to about 20 μM. In some cases, the desired concentration of the C5 inhibitor in subject plasma may be about 0.1 μg / mL to about 1000 μg / mL. In other cases, the desired concentration of the C5 inhibitor in subject plasma may be about 0.01 μg / mL to about 2 μg / mL, about 0.02 μg / mL to about 4 μg / mL, about 0.05 μg / mL to about 5 μg / mL, about 0.1 μg / mL to about 1.0 μg / mL, about 0.2 μg / mL to about 2.0 μg / mL, about 0.5 μg / mL to about 5 μg / mL, about 1 μg / mL to about 5 μg / mL, about 2 μg / mL to about 10 μg / mL, about 3 μg / mL to about 9 μg / mL, about 5 μg / mL to about 20 μg / mL, about 10 μg / mL to about 40 μg / mL, about 30 μg / mL to about 60 μg / mL, about 40 μg / mL to about 80 μg / mL, about 50 μg / mL to about 100 μg / mL, about 75 μg / mL to about 150 μg / mL, or at least 150 μg / mL. In other embodiments, the C5 inhibitor is administered at a dose sufficient to achieve a peak serum concentration (C max ) of at least 0.1 μg / mL, at least 0.5 μg / mL, at least 1 μg / mL, at least 5 μg / mL, at least 10 μg / mL, at least 50 μg / mL, at least 100 μg / mL, or at least 1000 μg / mL.

[0144] In some embodiments, a dose sufficient to maintain a C5 inhibitor level of about 0.1 μg / mL to about 20 μg / mL is provided to reduce hemolysis in a subject by about 25% to about 99%.

[0145] In some embodiments, the C5 inhibitor is administered daily at a dose sufficient to deliver approximately 0.1 mg / day to approximately 60 mg / day per kg of body weight of the subject. In some cases, the C5 achieved with each administration max The concentration ranges from approximately 0.1 μg / mL to approximately 1000 μg / mL. In such cases, the area under the curve (AUC) between doses is approximately 200 μg. * time / mL ~ approx. 10,000μg * Time / mL may also be acceptable.

[0146] According to some methods of the present invention, the C5 inhibitor of the present invention is provided at a concentration required to achieve the desired effect. In some cases, the compounds and compositions of the present invention are provided in an amount required to reduce a given reaction or process by half. The concentration required to achieve such reduction is referred herein as the 50% inhibitory concentration (half maximal inhibitory concentration) or "IC50% inhibitory concentration". 50 This is referred to as ". Alternatively, the compounds and compositions of the present invention may be provided in amounts required to half-maximal enhance a given reaction, activity, or process. The concentration required for such enhancement is referred to herein as half maximal effective concentration or "EC 50 It is referred to as "...".

[0147] The C5 inhibitor of the present invention may be presented in an amount of 0.1 to 95% by weight of the total weight of the composition. In some cases, the C5 inhibitor is provided by intravenous (IV) administration. In some cases, the C5 inhibitor is provided by subcutaneous (SC) administration.

[0148] SC administration of the C5 inhibitor of the present invention may, in some cases, offer advantages over IV administration. SC administration may enable patients to self-administer the treatment. Such treatment may be advantageous in that patients can administer the treatment to themselves at their own homes, avoiding the need to travel to a provider or medical facility. Furthermore, SC treatment may enable patients to avoid long-term complications associated with IV administration, such as infections, decreased venous access, local thrombosis, and hematomas. In some embodiments, SC treatment may enhance patient compliance, patient satisfaction, and quality of life, and reduce treatment costs and / or drug requirements.

[0149] In some cases, daily SC administration provides steady-state C5 inhibitor concentrations reached within 1-3 doses, 2-3 doses, 3-5 doses, or 5-10 doses. In some cases, a daily SC dose of 0.1 mg / kg can achieve sustained C5 inhibitor levels of 2.5 μg / mL or higher and / or inhibition of complement activity exceeding 90%.

[0150] The C5 inhibitors of the present invention may exhibit slow absorption pharmacokinetics (time to peak observation concentration exceeding 4-8 hours) and high bioavailability (approximately 75%-100%) after SC administration. In some embodiments, the dose and / or administration is determined by the half-life (t) of the C5 inhibitor level in the target or target body fluid (e.g., plasma). 1 / 2 ) is changed to adjust. In some cases, t 1 / 2This is at least 1 hour, at least 2 hours, at least 4 hours, at least 6 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 16 hours, at least 20 hours, at least 24 hours, at least 36 hours, at least 48 hours, at least 60 hours, at least 72 hours, at least 96 hours, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, or at least 16 weeks.

[0151] In some embodiments, the C5 inhibitor of the present invention has a long terminal t 1 / 2 This can show an extended terminal t. 1 / 2 This may be due to further target binding and / or additional plasma protein binding. In some cases, the C5 inhibitors of the present invention exhibit t(24 hours or more) in both plasma and whole blood. 1 / 2 The values ​​are shown. In some cases, C5 inhibitors do not lose their functional activity after incubation in human whole blood at 37°C for 16 hours.

[0152] In some embodiments, the dose and / or administration is modified to adjust the steady-state volume of distribution of the C5 inhibitor. In some cases, the steady-state volume of distribution of the C5 inhibitor is approximately 0.1 mL / kg to approximately 1 mL / kg, approximately 0.5 mL / kg to approximately 5 mL / kg, approximately 1 mL / kg to approximately 10 mL / kg, approximately 5 mL / kg to approximately 20 mL / kg, approximately 15 mL / kg to approximately 30 mL / kg, approximately 10 mL / kg to approximately 200 mL / kg, approximately 20 mL / kg to approximately 60 mL / kg, approximately 30 mL / kg to approximately 70 mL / kg, approximately 50 mL / kg to approximately 200 mL / kg, approximately 100 mL / kg to approximately 500 mL / kg, or at least 500 mL / kg. In some embodiments, the dose and / or administration of the C5 inhibitor is adjusted to ensure that the steady-state volume of distribution is equal to at least 50% of the total blood volume. In some embodiments, the distribution of the C5 inhibitor may be restricted to a plasma compartment.

[0153] In some embodiments, the C5 inhibitors of the present invention exhibit total clearance rates of approximately 0.001 mL / hour / kg to approximately 0.01 mL / hour / kg, approximately 0.005 mL / hour / kg to approximately 0.05 mL / hour / kg, approximately 0.01 mL / hour / kg to approximately 0.1 mL / hour / kg, approximately 0.05 mL / hour / kg to approximately 0.5 mL / hour / kg, approximately 0.1 mL / hour / kg to approximately 1 mL / hour / kg, approximately 0.5 mL / hour / kg to approximately 5 mL / hour / kg, approximately 0.04 mL / hour / kg to approximately 4 mL / hour / kg, approximately 1 mL / hour / kg to approximately 10 mL / hour / kg, approximately 5 mL / hour / kg to approximately 20 mL / hour / kg, approximately 15 mL / hour / kg to approximately 30 mL / hour / kg, or at least 30 mL / hour / kg.

[0154] The duration during which the highest concentration of the C5 inhibitor is maintained in the target (e.g., target serum) (T maxThe T(II, IV) values ​​may be adjusted by changing the dose and / or administration (e.g., subcutaneous administration). In some cases, C5 inhibitors are used for approximately 1 minute to 10 minutes, approximately 5 minutes to 20 minutes, approximately 15 minutes to 45 minutes, approximately 30 minutes to 60 minutes, approximately 45 minutes to 90 minutes, approximately 1 hour to 48 hours, approximately 2 hours to 10 hours, approximately 5 hours to 20 hours, approximately 10 hours to 60 hours, approximately 1 day to 4 days, approximately 2 days to 10 days, or at least 10 days. max It has a value.

[0155] In some embodiments, the C5 inhibitors of the present invention may be administered without off-target effects. In some cases, the C5 inhibitors of the present invention do not inhibit hERGs (human etheragogo-related genes) even at concentrations of 300 μM or less. SC injections of the C5 inhibitors of the present invention at dose levels up to 10 mg / kg are well tolerable and may not cause any cardiovascular adverse effects (e.g., increased risk of long-term ventricular repolarization) and / or respiratory adverse effects.

[0156] The dose of a C5 inhibitor may be determined using no-observed-adverse-effect levels (NOAELs) observed in other species. Such species may include, but are not limited to, monkeys, rats, rabbits, and mice. In some cases, the human equivalent dose (HED) may be determined by non-proportional scaling from NOAELs observed in other species. In some cases, the HED results in therapeutic limits of approximately 2-5 times, 4-12 times, 5-15 times, 10-30 times, or at least 30 times. In some cases, the therapeutic limit is determined by human C5 exposure and human C5 levels evaluated in humans. max It is determined by using the level.

[0157] In some embodiments, the C5 inhibitors of the present invention enable rapid drug-free periods in cases of infection where long-term inhibition of the complement system has been found to be harmful. The administration of C5 inhibitors described in this invention may be modified to reduce potential clinical risks to the subject. Infection with Neisseria meningitidis is a known risk of C5 inhibitors, including eculizumab. In some cases, the risk of infection with Neisseria meningitidis can be minimized by initiating one or more prophylactic steps. Such steps may include excluding subjects who may already be colonized by these bacteria. In some cases, the prophylactic steps may include the co-administration of one or more antibiotics. In some cases, ciprofloxacin may be co-administered. In some cases, ciprofloxacin may be administered orally co-administered at a dose of about 100 mg to about 1000 mg (e.g., 500 mg).

[0158] In some embodiments, C5 inhibitor administration may be carried out using an automated injection device. Such a device may enable self-administration (e.g., daily administration). Medication frequency In some embodiments, the C5 inhibitor of the present invention is administered hourly, every 2 hours, every 4 hours, every 6 hours, every 12 hours, every 18 hours, every 24 hours, every 36 hours, every 72 hours, every 84 hours, every 96 hours, every 5 days, every 7 days, every 10 days, every 14 days, weekly, every 2 weeks, every 3 weeks, every 4 weeks, monthly, every 2 months, every 3 months, every 4 months, every 5 months, every 6 months, annually, or at least annually. In some cases, the C5 inhibitor is administered once daily, or two, three, or more partial doses at appropriate intervals throughout the day.

[0159] In some embodiments, the C5 inhibitor is administered multiple times a day. In some cases, the C5 inhibitor is administered daily for 7 days. In some cases, the C5 inhibitor is administered daily for 7 to 100 days. In some cases, the C5 inhibitor is administered daily for at least 100 days. In some cases, the C5 inhibitor is administered daily indefinitely.

[0160] C5 inhibitors delivered intravenously may be delivered by prolonged infusions, for example, over 5, 10, 15, 20, or 25 minutes. Doses may be repeated, for example, regularly, for example, hourly, daily, weekly, every other week (i.e., every two weeks), for one month, two months, three months, four months, or more than four months. After the initial treatment plan, the therapeutic agent may be administered on a lower frequency basis. For example, after three months of every-other-week administration, doses may be repeated once every one month, six months, or one year or more. Administration of a C5 inhibitor may reduce, decrease, increase, or alter binding or any physiologically harmful processes (for example, in the patient's cells, tissues, blood, urine, or other compartments) by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% or more.

[0161] Prior to administration of the full dose of a C5 inhibitor and / or C5 inhibitor composition, the patient may be administered a smaller dose, for example 5% of the full dose, and monitored for adverse effects, such as allergic or infusion reactions, or elevated lipid levels or blood pressure. In another example, the patient may be monitored for undesirable immunostimulatory effects, such as elevated cytokine levels (e.g., TNF-α, Il-1, Il-6, or Il-10).

[0162] Genetic predisposition plays a role in the development of certain diseases or disorders. Therefore, patients requiring C5 inhibitors may be identified by taking a family history or, for example, by screening for one or more genetic markers or variants. Before prescribing or administering the therapeutic composition of the present invention, healthcare providers, such as physicians, nurses, or family members, may analyze the family history.

[0163] III. Kit Any of the C5 inhibitors described herein may be provided as part of a kit. In non-limiting cases, the C5 inhibitor may be included in a kit for treating a disease. The kit may include a vial of sterile, dried C5 inhibitor powder, a sterile solution for dissolving the dried powder, and a syringe for an infusion set for administering the C5 inhibitor.

[0164] When the C5 inhibitor is provided as a dry powder, it is considered that an amount between 10 micrograms and 1000 milligrams, or at least or more, of the C5 inhibitor may be provided in the kit of the present invention.

[0165] A typical kit may include at least one vial, test tube, flask, bottle, syringe, and / or other container or device containing, and preferably suitably, a C5 inhibitor formulation. The kit may also include one or more secondary containers containing sterile, pharmaceutically acceptable buffers and / or other diluents.

[0166] In some embodiments, the compounds or compositions of the present invention are provided in a borosilicate vial. Such a vial may include a cap (e.g., a rubber stopper). In some cases, the cap includes a rubber stopper coated with FLUROTEC®. The cap may be secured in place by an overseal, but not limited to an aluminum flip-off overseal.

[0167] The kit may further include instructions for using the kit components, as well as instructions for using any other reagents not included in the kit. The instructions may include feasible variations.

[0168] IV. Definition Bioavailability: As used herein, the term “bioavailability” refers to the systemic availability of a given amount of a compound (e.g., a C5 inhibitor) administered to a subject. Bioavailability is the area under the curve (AUC) or peak serum or plasma concentration (C5) of the unchanged form of the compound after administration to the subject. max AUC can be evaluated by measuring the area under the curve. AUC is a measure of the area under the curve when the serum or plasma concentration of a compound is plotted on the y-axis against time on the x-axis. Generally, the AUC for a particular compound can be calculated using methods known to those skilled in the art and / or methods described in GS Banker, Modern Pharmaceutics, Drugs and the Pharmaceutical Sciences, Vol. 72, Marcel Dekker, New York, Inc., 1996 (the contents of which are incorporated herein by reference in their entirety).

[0169] Biosystem: As used herein, the term “biological system” refers to a cell, cell compartment, cell, cell culture, tissue, organ, organ system, organism, multicellular organism, body fluid, or any biological entity that performs at least one biological function or task within a cell membrane, cell compartment, cell, cell culture, tissue, organ, organ system, organelle, body fluid, biological signaling pathway (e.g., receptor-activated signaling pathway, charge-activated signaling pathway, metabolic pathway, cellular signaling pathway, etc.), group of proteins, group of nucleic acids, or group of molecules (including, but not limited to, biomolecules). In some embodiments, a biological system is a cellular signaling pathway that includes intracellular and / or extracellular signaling biomolecules. In some embodiments, a biological system includes a proteolytic cascade (e.g., a complement cascade).

[0170] Buffering agents: As used herein, the term “buffering agent” refers to a compound used in a solution intended to resist changes in pH. Such compounds include, but are not limited to, acetic acid, adipic acid, sodium acetate, benzoic acid, citric acid, sodium benzoate, maleic acid, sodium phosphate, tartaric acid, lactic acid, potassium metaphosphate, glycine, sodium bicarbonate, potassium phosphate, sodium citrate, and sodium tartrate.

[0171] Clearance rate: As used herein, the term “clearance rate” refers to the rate at which a particular compound is removed from a biological system or bodily fluid. Compound: As used herein, the term “compound” refers to a characteristic chemical entity. In some embodiments, a particular compound may exist in one or more isomeric or isotopic forms (including, but not limited to, stereoisomers, geometric isomers, and isotopes). In some embodiments, a compound is provided or used only in a single such form. In some embodiments, a compound is provided or used as a mixture of two or more such forms (including, but not limited to, racemic mixtures of stereoisomers). Those skilled in the art will understand that some compounds exist in different forms and exhibit different properties and / or activities (including, but not limited to, biological activity). In such cases, in use according to the present invention, selecting or avoiding particular forms of the compound is within the scope of those skilled in the art. For example, a compound containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms.

[0172] Cyclic or Cyclized: As used herein, the term “cyclic” refers to the presence of a continuous loop. A cyclic molecule does not need to be cyclic only when linked together to form an indestructible chain of subunits. A cyclic polypeptide may contain a “cyclic loop” formed when two amino acids are linked by a crosslinking site. The cyclic loop contains amino acids along the polypeptide present between the crosslinked amino acids. The cyclic loop may contain 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids.

[0173] Downstream Events: As used herein, the terms “downstream” or “downstream event” refer to any event that occurs after and / or as a result of another event. In some cases, downstream events are events that occur after or as a result of C5 cleavage and / or complement activation. Such events may include, but are not limited to, the production of C5 cleavage products, MAC activation, hemolysis, and hemolysis-related disorders (e.g., PNH).

[0174] Equilibrium dissociation constant: As used herein, the term "equilibrium dissociation constant" or "K D " refers to a value that indicates the tendency for two or more drugs (for example, two proteins) to reversibly separate. In some cases, K D This indicates the concentration of the primary drug when half of the total levels of the secondary drug are related to the primary drug.

[0175] Half-life: Where used herein, the term "half-life" or "t 1 / 2 "Terminal half-life" refers to the time it takes for a given process or compound concentration to reach half of its final value. 1 / 2 This refers to the time it takes for the plasma concentration of a factor to decrease by half after the factor's concentration reaches a false equilibrium.

[0176] Hemolysis: As used herein, the term "hemolysis" refers to the destruction of red blood cells. Identity: As used herein, the term “identity” refers to the relationship between sequences by comparison when referring to polypeptides or nucleic acids. The term is used to describe the degree of sequence relevance between polymer sequences and may include percentages of monomeric components that conform to gap alignments (if any) processed by a particular mathematical model or computer program (i.e., “algorithm”). The identity of related polypeptides can be readily calculated by known methods. Such methods include, but are not limited to, those described by others (Lesk, AM, ed., “Computational Molecular Biology,” Oxford University Press). University Press, New York, 1988; Smith, DW (ed.), "Biocomputing: Informatics and Genome Projects," Academic Press, New York, 1993; Griffin, AM et al. (eds.), "Computer Analysis of Sequence Data This includes previously published works in: "Sequence Data)", Part 1, Humana Press, New Jersey, 1994; von Heinje, G., "Sequence Analysis in Molecular Biology", Academic Press, 1987; Gribskov, M. et al., eds., "Sequence Analysis Primer", M. Stockton Press, New York, 1991; and Carillo et al., Applied Mathematics, SIAM J, 1988, Vol. 48, p. 1073).

[0177] Inhibitor: As used herein, the term “inhibitor” refers to any agent that blocks or reduces the development of a specific event; cellular signaling; chemical pathway; enzymatic reaction; cellular process; interaction between two or more entities; biological event; disease; disorder; or pathological condition.

[0178] Intravenous: As used herein, the term “intravenous” refers to the region inside a blood vessel. Intravenous administration typically refers to the delivery of a compound into the bloodstream via injection into a blood vessel (e.g., a vein).

[0179] in vitro: As used herein, the term “in vitro” refers to an event that occurs in an artificial environment (e.g., in a test tube or reaction vessel, under cell culture conditions, in a petri dish, etc.) rather than inside a living organism (e.g., an animal, plant, or microorganism).

[0180] in vivo: As used herein, the term “in vivo” refers to an event occurring within an organism (for example, an animal, plant, or microorganism, or its cells or tissues).

[0181] Lactam crosslinking: As used herein, the term “lactam crosslinking” refers to amide bonds that form crosslinks between chemical groups in a molecule. In some cases, lactam crosslinking is formed between amino acids in a polypeptide.

[0182] Linker: The term "linker," as used herein, refers to a group of atoms (e.g., 10 to 1,000 atoms), a molecule, or other compound used to link two or more entities. A linker may link such entities via covalent or non-covalent (e.g., ionic or hydrophobic) interactions. A linker may contain a chain of two or more polyethylene glycol (PEG) units. In some cases, a linker may be cleavable.

[0183] Ventilation per minute: As used herein, the term "ventilation per minute" refers to the volume of air inhaled or exhaled from the lungs of the subject per minute. Non-protein composition: As used herein, the term "non-protein composition" refers to a composition containing any non-natural proteins, such as non-natural components, such as non-natural amino acids.

[0184] Patient: As used herein, “patient” means a person who seeks or may need treatment, who needs treatment, who is receiving treatment, who will receive treatment, or who is under the care of a specialist skilled in a particular disease or condition.

[0185] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” means a composition comprising at least one active ingredient (e.g., a C5 inhibitor) in a form and amount that allows the active ingredient to be therapeutically effective.

[0186] Pharmacopoeia-acceptable: The term "pharmacopoeia-acceptable" is used herein to mean a compound, material, composition, and / or dosage form that is suitable for use in contact with human and animal tissues, free from excessive toxicity, irritation, allergic reactions, or other problems or complications, balanced by a reasonable risk-benefit ratio, and within the bounds of sound medical judgment.

[0187] pharmaceutically acceptable excipients: The term “pharmaceutically acceptable excipients,” as used herein, refers to any component present in a pharmaceutical composition other than the active agent (e.g., R5000 or its variants) that is substantially non-toxic and non-inflammatory in the patient. In some embodiments, a pharmaceutically acceptable excipient is a solvent capable of suspending or dissolving the active agent. Excipients may include, for example, anti-adhesives, antioxidants, binders, coatings, compression aids, disintegrants, pigments (colorants), emollients, emulsifiers, fillers (diluents), film-forming agents or coatings, flavorings, aromas, lubricants (flow enhancers), preservatives, printing inks, adsorbents, suspending agents or dispersants, sweeteners, and hydration water. Examples of excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, calcium (II) phosphate, calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0188] Plasma compartment: As used herein, the term "plasma compartment" refers to the intravascular space occupied by plasma. Salt: As used herein, the term “salt” refers to a compound consisting of an anion bonded to a cation. Such compounds may include sodium chloride (NaCl) or, but are not limited to, other classes of salts including acetates, chlorides, carbonates, cyanides, nitrites, nitrates, sulfates, and phosphates.

[0189] Sample: As used herein, the term “sample” refers to a certain volume or portion taken from a source and / or provided for analysis or processing. In some embodiments, a sample is derived from a biological source such as tissue, cells, or components (body fluids including, but not limited to, blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, amniotic fluid, amniotic blood, urine, vaginal fluid, and semen). In some embodiments, a sample may be or contain homogenates, solubilizers, or extracts (prepared from, but not limited to, a whole organism or a subset or fraction or portion thereof of its tissues, cells, or components, such as plasma, serum, cerebrospinal fluid, lymph, skin, airways, intestines, and external sections of the genitourinary tract, tears, saliva, milk, blood cells, tumors, or organs). In some embodiments, a sample is or contains a culture medium, such as a nutrient broth or gel, which may contain cellular components, such as proteins. In some embodiments, the “primary” sample is a certain volume of the source. In some embodiments, the primary sample may undergo one or more processing steps (e.g., separation, purification, etc.) to prepare a sample for analysis or other applications.

[0190] Subcutaneous: As used herein, the term “subcutaneous” refers to the space beneath the skin. Subcutaneous administration is the delivery of a compound beneath the skin. Subject: As used herein, the term “subject” refers to any organism to which a compound according to the present invention may be administered, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, pig subjects, non-human primates, and humans).

[0191] Substantially: As used herein, the term “substantially” refers to a qualitative state that represents the total amount or degree or extent of the characteristics or properties of interest. Those skilled in the art in biology will understand that biological and chemical phenomena rarely, if they exist, reach completion and / or completeness, or achieve or avoid absolute results. Thus, the term “substantially” is used herein to capture the potential lack of completeness inherent in a number of biological and chemical phenomena.

[0192] Therapeutic dose: As used herein, the term “therapeutic dose” means the amount of drug to be delivered (e.g., a C5 inhibitor) that, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, is sufficient to treat, improve, diagnose, prevent, and / or delay the onset of a disease, disorder, and / or condition.

[0193] Tidal volume: As used herein, the term “tidal volume” refers to the normal lung volume of air exchanged between breaths (in the absence of any extra effort). T max :As used herein, the term "T max " " refers to the period during which the highest concentration of a compound in the subject or body fluid is maintained.

[0194] To treat: As used herein, the term “to treat” means to partially or completely alleviate, remit, improve, relieve, delay the onset of, inhibit the progression of, reduce the severity of, and / or decrease the incidence of one or more symptoms or features of a particular disease, disorder, and / or condition. Treatment may be applied to subjects who do not show signs of a disease, disorder, and / or condition, and / or subjects who show only the initial signs of a disease, disorder, and / or condition, with the aim of reducing the risk of developing pathology associated with the disease, disorder, and / or condition.

[0195] Volume of distribution: When used herein, the term “volume of distribution” or “V dist "Volume of distribution" refers to the volume of body fluid required to contain the total amount of a compound in the body at the same concentration as in blood or plasma. Volume of distribution can reflect the extent to which a compound is present in extravascular tissue. A large volume of distribution reflects the tendency of the compound to bind to tissue components compared to plasma protein components. In clinical situations, V dist By using this method, the loading dose of the compound can be determined, and the steady-state concentration of the compound can be achieved.

[0196] V. Equivalents and Range While various embodiments of the present invention have been specifically presented and described, it will be understood by those skilled in the art that various modifications to the form and details can be made herein without departing from the spirit and scope of the invention as defined in the appended claims.

[0197] Those skilled in the art will be able to identify, or confirm, many equivalent forms of the specific embodiments of the present invention described herein without going beyond ordinary experimentation. The scope of the present invention is not limited to the foregoing description, but is as specified in the appended claims.

[0198] In the claims, articles such as “a,” “an,” and “the” can mean one or more unless there is a conflicting indication or it is particularly obvious from the context. A claim or description containing “or” between one or more members of a group is considered satisfied if one, more than one, or all of the group members are present in, used in, or otherwise adapted to a given product or process, unless there is a conflicting indication or it is particularly obvious from the context. The present invention includes embodiments in which strictly one member of the group is present in, used in, or otherwise adapted to a given product or process. The present invention includes embodiments in which more than one or all of the group members are present in, used in, or otherwise adapted to a given product or process.

[0199] Furthermore, it should be noted that the term “comprising” is intended to be open and allows for, but does not require, the inclusion of additional elements or processes. Therefore, wherever the term “comprising” is used herein, the terms “consisting of” and “or including” are also implicitly disclosed.

[0200] If a range is given, the endpoints are included. Furthermore, unless otherwise specified or is particularly obvious from the context and the understanding of those skilled in the art, values ​​expressed as a range should be understood to mean any specific value or subrange within the specified range in various embodiments of the invention, up to 1 / 10th of the lower limit of the range, unless the context clearly distinguishes otherwise.

[0201] In addition, any particular embodiment of the present invention that is included in the prior art should be understood to be expressly excluded from any one or more claims. Since such embodiments are considered to be known to those skilled in the art, they may be excluded even if their exclusion is not explicitly stated herein. Any particular embodiment of a composition according to the present invention (e.g., any nucleic acid, a protein encoded thereby, any method of production, any method of use, etc.) may be excluded from any one or more claims for any reason, with or without the existence of prior art.

[0202] All cited sources, such as references, publications, databases, database entries, and technologies cited herein, are incorporated by reference to this application, even if not explicitly stated in the citation. In the event of any conflict between the content of a cited source and the content of this application, the content of this application shall prevail.

[0203] Sections and titles are not intended to be limiting. (Examples) Example 1. Preparation of R5000 aqueous solution Polypeptides were synthesized using a standard solid-phase Fmoc / tBu method. Synthesis was performed using a Liberty automated microwave peptide synthesizer (CEM, Matthews, North Carolina) with a standard protocol using Rink amide resin, although other automated synthesizers without microwave capabilities may be used. All amino acids were obtained from suppliers. The coupling reagent used was 2-(6-chloro-1-H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate (HCTU), and the base was diisopropylethylamine (DIEA). Polypeptides were cleaved from the resin over 3 hours using 95% TFA, 2.5% TIS, and 2.5% water, and isolated by precipitation with ether. Crude polypeptides were purified over 30 minutes using reverse-phase preparative HPLC with a C18 column and a 20% to 50% gradient of acetonitrile / water and 0.1% TFA. The fractions containing pure polypeptides were collected and freeze-dried, and all polypeptides were analyzed by LC-MS.

[0204] R5000 (SEQ ID NO: 1) was prepared as a cyclic peptide having 15 amino acids (four of which are non-natural amino acids), an acetylated N-terminus, and a C-terminal carboxylic acid. The C-terminal lysine of the core peptide has a modified side chain forming an N-ε-(PEG24-γ-glutamic acid-N-α-hexadecanoyl)lysine residue. This modified side chain contains a polyethylene glycol spacer (PEG24) attached to an L-γ-glutamic acid residue derivatized with a palmitoyl group. Cyclization of R5000 is via lactam crosslinking between the L-Lys1 and L-Asp6 side chains. All amino acids in R5000 are L-amino acids. R5000 has a molecular weight of 3562.23 g / mol and C 172 H 278 N 24 O 55 It has the chemical formula.

[0205] Similar to eculizumab, R5000 blocks the proteolytic cleavage of C5 into C5a and C5b. Unlike eculizumab, R5000 also binds to C5b and can block its binding to C6, thereby preventing subsequent MAC assembly.

[0206] R5000 was prepared as an aqueous solution for injection containing 40 mg / mL of R5000 in a formulation of 50 mM sodium phosphate and 75.7 mM sodium chloride at a pH of 7.0 ± 0.3.

[0207] Example 2. Administration and storage of R5000 R5000 is administered by subcutaneous (SC) or intravenous (IV) injection, and the dose (dosage) is adjusted based on the subject's body weight in mg / kg. This is obtained using a set of fixed doses adjusted for a set of body weight tiers. Overall, human administration supports a wide body weight range of 43–109 kg. Subjects with heavier body weights (>109 kg) should be handled on a case-by-case basis, taking medical monitoring into consideration.

[0208] R5000 should be stored at 2°C to 8°C [36°C to 46°F]. Once dispensed to the target, R5000 should be stored at a controlled room temperature (20°C to 25°C [68°F to 77°F]) for up to 30 days, protected from sources of excessive temperature fluctuations such as high heat or exposure to light. Storage of R5000 at temperatures other than room temperature should preferably be avoided. R5000 may be stored under these conditions for up to 30 days.

[0209] Example 3. Stability Test Stability testing will be conducted in accordance with International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) Q1A, "Stability of New Drug Substances and Products." Samples from the aqueous solution of Example 1 will be held at three temperatures: -20°C, 5°C, and 25°C. Test intervals will be every three months after one month, two months, and three months, up to a maximum of 24 months. Samples will be tested for appearance (e.g., clarity, color, presence of precipitate), pH, osmotic pressure, concentration, purity, target activity (e.g., by RBC solubility assay), particulate matter level, endotoxin level, and sterility. A sample is considered stable if, at each of the temperature conditions tested, it has a clear, colorless appearance without visible particles; a pH of 7 ± 0.3; an osmotic pressure of 260–340 mOsm / kg; a purity of ≥95% (and no single impurities >3%); target activity comparable to the reference standard; a particle level of ≤6,000 particles / vial for ≥10 μm particles and ≤600 particles / vial for ≥25 μm particles; an endotoxin level of ≤100 EU / mL; and no microbial growth.

[0210] Example 4. Freeze-thaw stability Tests were conducted to examine the stability of the aqueous solution of Example 1 when exposed to multiple freeze-thaw cycles. R5000 showed no degradation or other changes after 5 freeze-thaw cycles.

[0211] Example 5. Evaluation of coupling based on surface plasmon resonance (SPR). The bond interaction between R5000 and C5 was measured using surface plasmon resonance. R5000 has an equilibrium dissociation constant of 0.42 nM at 25°C (K D )(n=3) and K at 0.78 nM at 37°C D (n=3) bonded to C5. Overall surface plasmon resonance data, combined with high-resolution cocrystal structure analysis, indicates that R5000 exhibits a specific, strong, and rapid association with C5 and a slow dissociation rate.

[0212] Example 6. Evaluation of C5 cleavage inhibition R5000 was evaluated for its inhibition of C5 cleavage to C5a and C5b. The inhibitory activity of R5000 against host C5 is an important factor in selecting an animal model suitable for drug safety. Inhibition of C5 cleavage is the basis for the clinical efficacy of eculizumab, the only currently approved therapeutic agent for the treatment of PNH. R5000 showed dose-dependent inhibition of C5a formation following activation of the classical pathway (IC 50 = 4.8 nM; Figure 1) and dose-dependent inhibition of C5b (measured as formation of C5b-9 or MAC) upon activation of the classical and alternative complement pathways (IC 50 = 5.1 nM; Figure 2).

[0213] Example 7. Inhibition of Complement-Induced Red Blood Cell (RBC) Hemolysis The RBC lysis assay is a reliable method for screening complement inhibitors in serum / plasma from various species and comparing the relative activities of test substances. To evaluate the inhibitory activity of peptides containing R5000 against complement function in several species, an in vitro functional assay was used. In this assay, the ability of complement components of the classical pathway to lyse sheep RBCs pre-coated with rabbit anti-sheep RBC antibody was tested. When antibody-coated RBCs are incubated with test serum, the classical pathway of complement is activated, and the hemolysis result is monitored by the release of hemoglobin. Antibody-sensitized sheep erythrocytes were used as a vehicle for lysis in this assay, and sera and / or plasma from various species were used at their predetermined 50% hemolytic complement activity (CH 50 ). [[ID=I4]]

[0214] R5000 showed potent inhibition of complement-induced RBC hemolysis in human, non-human primate, and porcine sera and / or plasma (see the following table).

[0215] [Table 1]

[0216] Weak activity was observed in rat plasma (>100 times lower than in cynomolgus monkeys), and little activity was observed in other rodents, dogs, or rabbits. Structural data obtained from co-crystallization of human C5 with molecules closely related to R5000, through careful analysis of the primary amino acid sequence at the drug binding site of the target protein, provide an explanation for this type of selectivity. While the primate sequence was 100% conserved within the residues involved in R5000 interaction, significant differences existed within these residues in rodents, particularly dogs, when identical portions of the protein were absent. These amino acid differences were sufficient to explain the heterologous activity characteristics of R5000.

[0217] The ability of R5000 to inhibit complement-mediated erythrocyte lysis via classical and alternative complement activation pathways was also tested. The classical pathway was evaluated using two different assays with antibody-sensitized sheep erythrocytes. In one method, hemolysis was evaluated using 1% normal human serum, while in the second assay, 1.5% C5-depleted human serum containing 0.5 nM human C5 was used. Inhibition of the alternative complement activation pathway was evaluated using rabbit erythrocytes in 6% normal human serum in the absence of calcium (see table below).

[0218] [Table 2]

[0219] R5000 exhibited complement-mediated lysis in both classical and alternative pathway assays. Example 8. Pharmacodynamics in cynomolgus monkeys Since R5000 is a potent complement inhibitor in primates, cynomolgus monkeys were selected for a multi-dose study to evaluate the inhibitory activity of R5000 in an animal model. Plasma drug concentrations were measured by LC-MS, and complement activity was assayed using the RBC lysis assay described in previous examples. Overall results from these studies indicated that a plasma drug concentration of 2.5 μg / mL or higher was required in monkeys to achieve >90% inhibition of complement activity (see Figure 3).

[0220] In a 7-day study, R5000 was administered to cynomolgus monkeys multiple times daily via subcutaneous injection (SC). Blood samples were analyzed for hemolysis as an indicator of complement activity at specified times (days 1, 4, and 7 after the initial dose, with data reported before administration each day) using an ex vivo sheep RBC lysis assay (using 1% plasma in the assay). Drug concentrations were measured from the same samples using an R5000-specific LC-MS method. As shown in the table below and Figures 4A and 4B, when R5000 was administered daily at either 0.21 or 4.2 mg / kg for 7 days, the lowest complement activity (<3% of the previous dose) was observed throughout the entire administration period.

[0221] [Table 3]

[0222] In ex vivo assays, hemolysis in the 0.21 mg / kg group was maintained at less than 90% of baseline after the initial dose, throughout the entire administration period, and for up to 24 hours after the final dose. After discontinuation of treatment, an increase in hemolysis levels was observed. Four days after the final dose (264 hours in Figure 4A), hemolysis was >75% of baseline. This correlates well with the measured plasma levels of the compound during and after administration (dotted line in Figure 4A). In the second animal group in the multiple-dose study, R5000 was administered daily at a dose of 4.2 mg / kg. In this group, hemolysis was essentially completely inhibited (<1%) throughout the entire administration period and maintained at less than 3% for 48 hours after the final dose (day 9; 216 hours in Figure 4B). Four days after the final dose (264 hours in Figure 4B), hemolysis reached approximately 10% of baseline. These results also correlated with plasma drug concentrations, showing suppression of complement activity throughout the entire administration period (compared to the results of the previous administration), and demonstrated an excellent correlation between pharmacokinetic and pharmacodynamic values.

[0223] The complement inhibitory activity of R5000 was evaluated in a 28-day repeated-dose study in cynomolgus monkeys using an ex vivo RBC hemolysis assay. R5000 was administered daily by subcutaneous injection at doses of 0, 1, 2, or 4 mg / kg / day for 28 days (Day 1: Figure 7, and Day 28: Figure 8). Results showed complete inhibition of hemolysis from 2 hours after the initial dose to Day 28, with hemolysis percentages <5% in the 1, 2, and 4 mg / kg / day groups compared to >90% in the control group. After a 28-day recovery period, sample values ​​returned to near baseline hemolysis levels, and little complement inhibition was observed. The absence of complement inhibitory activity at the end of the recovery period indicated drug clearance from the animals.

[0224] Complement inhibition was also tested as part of a 13-week repeated-dose study in cynomolgus monkeys. Monkey samples were analyzed using an ex vivo RBC hemolysis assay. R5000 was administered daily via subcutaneous injection at doses of 0, 0.25, 1, 2, or 10 mg / kg / day for 13 weeks. Similar to the 28-day study, results from the 13-week study showed complete ex vivo inhibition of hemolysis from 2 hours after the initial dose to 13 weeks of administration, with hemolysis percentages <5% in the 0.25, 1, 2, and 10 mg / kg / day groups compared to >90% in the control group. After a 28-day recovery period, sample values ​​returned to near baseline hemolysis levels, and little complement inhibition was observed. The absence of complement inhibitory activity at the end of the recovery period indicated clearance of the drug from the animals.

[0225] Example 9. Safety pharmacology When R5000 was administered to cynomolgus monkeys, no adverse effects on cardiovascular, respiratory, or central nervous system parameters were observed. Safety pharmacological parameters were evaluated in vitro using a human etheragogo-related gene (hERG) assay, and in vivo in monkeys, for cardiovascular, respiratory, and CNS parameters. CNS safety pharmacological evaluation was performed as part of a 28-day non-human primate toxicity study. A summary of the safety pharmacological studies using R5000 is shown in the table below.

[0226]

Table 4

[0227] The in vitro effect of R5000 on the cloned hERG potassium channel current (surrogate for IKr, rapidly activating delayed rectifier cardiac potassium current) expressed in human embryonic kidney 293 cells was evaluated using a parallel patch clamp system. The highest test concentration (300 μM) did not result in more than 50% hERG inhibition. Therefore, the IC 50 for R5000 was estimated to be greater than 300 μM (1.07 mg / mL).

[0228] In vivo cardiovascular and respiratory safety pharmacology studies were performed in conscious male cynomolgus monkeys. No deaths or significant clinical events were observed after administration of R5000. R5000-related effects on morphology and complete interbeat intervals were not seen at any R5000 dose (2 or 10 mg / kg on days 1 and 8). Only normal circadian variations were observed in electrocardiograms and body temperature (comparable to readings with vehicle treatment). Furthermore, no changes in heart rate and arterial pressure attributable to R5000 were observed at doses up to 10 mg / kg and plasma drug concentrations up to 79.1 μg / mL.

[0229] After treatment with R5000 at 2 or 10 mg / kg, no changes were observed in any of the respiratory parameters (respiratory rate, tidal volume, and minute ventilation) compared to values obtained after pretreatment or vehicle administration.

[0230] R5000 was administered daily to cynomolgus monkeys at 1, 2, or 4 mg / kg / day via subcutaneous injection to test its effect on the central nervous system (CNS). Parameters for evaluation included general attitude, behavior, motor function, cranial nerves, proprioceptive sensation, postural responses, and spinal nerves. No neurological changes were observed after treatment with R5000.

[0231] In conclusion, R5000 has a C content of 79.1 μg / mL. max Subcutaneous (SC) injection at dose levels up to 10 mg / kg was well-tolerated, and no adverse cardiovascular (without increased risk of QT prolongation or measurement of delayed ventricular repolarization), respiratory, or central nervous system effects occurred in conscious cynomolgus monkeys.

[0232] Example 10. Pharmacokinetics and drug metabolism in animals The table below lists the tests used to evaluate the in vitro and in vivo absorption, distribution, metabolism, and excretion of R5000. In the table below, CYP refers to cytochrome P450 enzymes, and UGT refers to UDP-glucuronosyltransferase enzymes.

[0233] [Table 5]

[0234] R5000 is extremely stable in vitro in rat, monkey, and human plasma, however, its pharmacokinetic properties after intravenous (IV) and SC administration in monkeys differed from those in rats (Figure 5A). The slow elimination kinetics observed in monkeys were primarily driven by high affinity interactions with the target protein C5 and other plasma proteins (e.g., albumin). The lack of specific target binding in rats resulted in more rapid elimination of R5000, with a terminal elimination time of 4-5 hours compared to >3 days in monkeys. 1 / 2 This was reflected in [the document].

[0235] Overall, preclinical data showed a high bioavailability (>75%) of R5000 after subcutaneous administration. In monkeys, the peak blood concentration (t max This is achieved between 8 and 16 hours after SC administration, indicating relatively slow absorption from the subcutaneous space. Aggregate data, including volume of distribution, high plasma protein binding, and distribution to plasma compartments in whole blood, indicate that R5000 is primarily confined to the plasma space and has little distribution to tissues.

[0236] absorption Pharmacokinetic (PK) studies were conducted in rats (single dose) and cynomolgus monkeys (single and multiple doses) using R5000 in a phosphate-buffered saline solution (pH 7.0).

[0237] In the rat study, a single dose of R5000 was administered subcutaneously to male Sprague-Dolly rats (n=3) at a dose of 1 mg / kg or 10 mg / kg. The measured pharmacokinetic (PK) parameters were as follows: max (maximum plasma drug concentration), T max (time required to reach peak plasma concentration after drug administration), t 1 / 2 (Half-life), AUC 0-last (Area under the plasma concentration-time curve between the first and final doses), and AUC 0-∞ This includes the area under the plasma concentration-time curve from time 0 to infinity. The results are summarized in the table below.

[0238] [Table 6]

[0239] Average AUC for both 1 mg / kg and 10 mg / kg 0-last The values ​​suggest exposure proportional to the dose. In primate studies, pharmacokinetic analysis was performed in cynomolgus monkeys after a single intravenous or subcutaneous administration of 0.4 or 0.5 mg / kg. The measured pharmacokinetic (PK) parameters were clearance (CL) and V z (distribution volume), V ss (Apparent volume of distribution in steady state), C max (maximum plasma drug concentration), T max (time required to reach peak plasma concentration after drug administration), t 1 / 2 (Half-life), AUC 0-last (Area under the drug concentration-time curve between the first and last dose), AUC 0-∞ The data includes the area under the drug concentration-time curve from time 0 to infinity, and %F (fraction). The results are shown in the table below (NA indicates inapplicability).

[0240] [Table 7]

[0241] As a result of a single 0.4 mg / kg SC administration, plasma exposure (AUC) of R5000 after IV and SC administration was observed. last ) are 429,638 and 325,317ng respectively. * The value was time / mL. The highest plasma concentration of R5000 after IV and SC administration (C max The values ​​were 4,745.5 and 2,490 ng / mL, respectively, and the T after SC administration max The duration was 8 hours. The subcutaneous bioavailability at 0.4 mg / kg was determined to be 75.7%. 1 / 2 The terminal phase in IV administration was 182.5 hours and 177.5 hours for IV and SC, respectively. z The mean distribution volume related to ) and clearance (CL) was determined to be 175.5 mL / kg and 0.011 mL / min / kg, respectively. This characteristic does not indicate clear binding based on in vitro activity testing, and therefore, the t of R5000 1 / 2 This is in contrast to rats, where the incubation period was 4-5 hours (see Figure 5A).

[0242] The repeated-dose pharmacokinetic study in monkeys included two subcutaneous dose levels, 0.21 and 4.2 mg / kg, administered daily for 7 days, with PK assessed daily and for 14 days after the final dose. In the multiple-dose study conducted in monkeys, C max The concentration increased with subsequent doses, reaching a steady-state peak and then a trough drug concentration (after 2-3 doses; see Figures 4A, 4B, and 5B). Plasma concentrations in the 0.2 and 4 mg / kg dose groups were 2,615 and 51,700 ng / mL, respectively, after the initial dose, with a mean C12. max It reached C max The mean C increased in both groups, and each consecutive dose was due to the long half-life of the molecule. By the fourth dose, the mean C in the 0.21 and 4.2 mg / kg dose groups was higher. maxThese values ​​were 5,305 and 68,750 ng / mL, which were 2.0 and 1.3 times the initial dose, respectively.

[0243] Overall, absorption could be characterized by slow absorption from the subcutaneous space and high bioavailability with subcutaneous administration. distribution In vitro plasma protein binding was >99.9% in human, rat, and monkey plasma, as measured by equilibrium dialysis at drug concentrations of 10 and 100 μM. High protein binding and limited distribution volume suggest that R5000 may be confined primarily to the plasma compartment and not readily distribute to the perivascular space.

[0244] blood distribution The ratio of drug distribution between plasma and red blood cells is an important parameter required to evaluate the pharmacokinetic properties of a drug, and was therefore calculated (see table below). In the table below, RBC represents red blood cells, P represents plasma, and WB represents whole blood.

[0245] [Table 8]

[0246] In whole blood partition assays, R5000 was found to be primarily present in the plasma fraction and did not show a significant distribution in the red blood cell fraction. Pharmacokinetics and drug interactions Cyclosporine (CsA) is a commonly administered drug in patients with paroxysmal nocturnal hemoglobinuria (PNH). Since R5000 is likely to be administered concurrently with CsA in PNH patients enrolled in planned clinical trials, the potential for drug-drug interactions between R5000 and CsA was evaluated in cynomolgus monkeys.

[0247] R5000 (2 mg / kg, subcutaneous, single dose) and cyclosporine A (CsA) (15 mg / kg, subcutaneous, single dose) were administered independently or together to two male monkeys, and plasma levels were evaluated using LC-MS / MS. No significant changes were observed in plasma exposure to either drug, indicating a low likelihood of drug-drug interaction (see table below). In the table below, C max The peak plasma drug concentration is shown, AUC is the area under the plasma concentration-time curve, and "a" indicates the difference between adjacent exposures, showing the ratio of exposure for R5000 + cyclosporine / R5000. "b" indicates the difference between adjacent exposures, showing the ratio of exposure for cyclosporine + R5000 / cyclosporine.

[0248] [Table 9]

[0249] No changes were observed in serum chemical parameters, including bilirubin (an endogenous substrate of OATP1 and OATP1B3), indicating no additive effect of CsA and R5000 on these transporters. In short, co-administration of CsA and R5000 has a low potential for drug-drug interaction, showed no effect on serum chemical parameters at plasma levels close to or exceeding those expected in clinical use, and demonstrated good tolerability.

[0250] Example 11. Pharmacokinetic / pharmacodynamic modeling and simulation of human pharmacokinetics The PK / PD model was constructed in silico using in vivo data obtained in cynomolgus monkeys. Model fit and accuracy were evaluated by comparing simulation results with newly generated experimental data. Once confirmed in monkeys, the final model was used to predict human pharmacokinetics by applying non-proportional scaling to its parameters. The resulting simulations support a planned dosing interval of once daily or less frequently in humans, and a daily dose of 0.1 mg / kg that maintains nearly 90% target inhibition at steady state (see Figure 6). Due to the long half-life of R5000, multiple doses are required to reach the final peak and trough drug concentrations. Plasma C max It is expected that when the drug concentration reaches a steady state after one week of daily administration, it will be approximately three times higher than the initial dose.

[0251] Example 12. Efficacy in humans: Phase I clinical trial design To evaluate the safety and pharmacokinetics of R5000, randomized, placebo-controlled, double-blind, single-escalation dose and multi-dose studies were conducted in healthy volunteers aged 18–65 years (excluding children and the elderly). In Part 1 of the study, cohorts of subjects were isolated and administered either a single-escalation dose (SAD) of R5000 or placebo. In Part 2 of the study, the multi-dose cohort (MD) was administered 0.2 mg / kg of R5000 (n=4) or placebo (n=2) daily for 7 days. All doses of R5000 were administered by subcutaneous injection at a dose determined by the cohort's dose requirements and the subject's body weight. Pregnant or lactating subjects, as well as any subjects with systemic infection or colonization of Neisseria meningitides, were excluded. Furthermore, all subjects received prophylaxis with ciprofloxacin, and subjects in the highest single-dose cohort (i.e., 0.4 mg / kg) as well as subjects in the multi-dose cohort received vaccination against Neisseria meningitides for at least 14 days prior to the study.

[0252] A total of 22 subjects were enrolled in a single-dose cohort study (n=14), of which two received R5000 at 0.05 mg / kg, and four each received 0.10, 0.20, and 0.40 mg / kg. These doses were selected using evaluated safety margins in humans (see previous studies and the table below). In the table below, C max This indicates the highest plasma drug concentration, AUC 0-last This line shows the area under the plasma concentration-time curve between the first and final doses.

[0253] [Table 10]

[0254] The initial dose of 0.05 mg / kg is far below 1 / 10 of the estimated human equivalent dose (HED). This dose is considered appropriate because no significant complement inhibition was expected at this dose. The exposure at the final NOAEL (day 28) in monkeys exceeds the predicted systemic exposure following the highest single SC dose of 0.8 mg / kg proposed in the trial.

[0255] Six subjects were enrolled in a multi-dose cohort; four received R5000 (0.2 mg / kg) and two received a placebo. Example 13. Treatment of a patient with PNH Patients with PNH are treated with R5000 at an effective dose of 0.1 mg / kg / day to 40 mg / kg / day. In these patients, complement inhibition is observed in more than 90% of cases. max The concentration reaches 3.1 μg / mL.

[0256] Example 14. Clinical trial of multiple doses of R5000 A phase 1 multi-dose clinical pharmacology study was conducted in healthy human volunteers to evaluate the safety, tolerability, pharmacokinetics, and pharmacokinetic and pharmacodynamic properties of R5000 after once-daily subcutaneous (SC) injection over 7 days. The study was single-center, randomized, double-blind, and placebo (PBO) controlled. Subjects were housed in a clinical pharmacology unit and received daily SC doses of 0.2 mg / kg of R5000 or a suitable PBO for 7 days. Safety was assessed by intensive clinical monitoring, and daily blood samples were taken immediately before administration and at 3 and 6 hours after administration each day to measure R5000 concentration and its ability to inhibit complement-mediated RBC lysis in an ex vivo antibody-sensitized sheep erythrocyte hemolysis assay by liquid chromatography / high-resolution mass spectrometry.

[0257] A total of six subjects were enrolled in the trial (four received the R5000 and two received the PBO). The demographics of the subjects are shown in the table below.

[0258] [Table 11]

[0259] As shown in the table below and the related Figure 9A (showing percentage hemolysis and plasma concentrations over 7 days), plasma concentrations showed a stable increase in exposure over the 7 days of administration. From these data, the half-life of R5000 was determined to be 7 days. Plasma levels returned to approximately 2000 ng / ml by day 15 and to approximately 1000 ng / ml by day 21 (Figure 9B).

[0260] [Table 12]

[0261] The pharmacokinetic (PK) parameters of R5000 after multiple dose SC administration (0.2 mg / kg / day) over 7 days are shown in the table below. The measured pharmacokinetic (PK) parameters are clearance (CL) and C. max (maximum plasma drug concentration), T max(Time to reach maximum plasma concentration after drug administration), t 1 / 2 (Half-life), AUC tau (Area under the plasma concentration-time curve from time 0 to 24 hours), AUC 0-inf (Area under the plasma concentration-time curve from time 0 to infinity), V z / F(Apparent volume of distribution), K el (Elimination rate), and F(Fraction) are included.

[0262]

Table 13

[0263] Mean C per day max and AUC tau were 2533 ng / mL and 50,010 ng * hours / mL, respectively, and were consistent with the results from the 0.2 mg / kg single-dose cohort over the same post-dose period. After 7 days of daily SC administration, C max and AUC tau increased by approximately 2.9-fold (mean 7-day C max = 7290 ng / mL) and approximately 3.0-fold (mean 7-day AUC tau = 151,300 ng * hours / mL), respectively. The median time to reach maximum plasma concentration on day 7 (T max ) was 3.0 hours, which was consistent with T max after single-dose SC administration (1-day T max median = 3.0 - 4.6 hours). This indicates consistent absorption rate of R5000 with repeated dosing. The 7-day mean of the apparent total body clearance of R5000 (7-day CL / F = 1.3 mL / hour / kg) was slightly increased compared to the total body clearance after single SC administration at 0.2 mg / kg [single ascending dose (SAD) was 0.2 mg / kg, CL / F = 0.29 mL / hour / kg]. However, the elimination rate constant (K el ) in R5000 was the same for single and repeated dosing (0.2 mg / kg SAD, mean K el = 0.0041 hour -1 ; 0.2 mg / kg MD, 7-day mean K el = 0.0043 hour -1) It was consistent later, which indicates that the clearance of R5000 did not significantly change with repeated dosing. The apparent volume of distribution (V z / F) of R5000 showed some increase with multiple-dose administrations of R5000 (SAD at 0.2 mg / kg, mean V z / F = 71.4 mL / kg; MD at 0.2 mg / kg, 7-day mean V z / F = 311.6 mL / kg). However, the 7-day V z / F in R5000 was still less than the total body water volume, which suggests that R5000 does not distribute into the extravascular space during repeated SC administration.

[0264]

Table 14

[0265] The mean percent inhibition compared to the baseline of hemolysis started at the starting point after administration on Day 1 and reached ≧95% 3 hours after administration, and continued throughout the 7-day administration (see the table below). All subjects showed a ≧90% reduction in hemolysis at all time points. It was observed that hemolysis was ≦3% in all subjects on Day 8 (24 hours after receiving the final dose). Hemolysis returned to the pre-dose level within 2 weeks after the final dose.

[0266] The study suggests that a low daily dose is suitable for achieving a steady-state level that completely and continuously inhibits complement and suppresses hemolysis. The study also suggests that once-weekly dosing may be sufficient to inhibit complement activity and reduce hemolysis in humans.

[0267] Complement activity in subject plasma samples was measured by Wieslab® ELISA (Euro Diagnostica, Malmö, Sweden) analysis. This assay measures alternative pathways for complement activation. Measurements via this assay indicated that the suppression of complement activity was rapid, complete, and persistent throughout the administration period in all subjects (see Figure 10A and the table below). In the table below, SEM values ​​show the mean standard error.

[0268] [Table 15]

[0269] On day 8 (24 hours after the final dose), complement activity was found to be ≤5% in all subjects. Complement activity returned to pre-dose levels within 2 weeks after the final dose (Figure 10B). R5000 was safe and well-tolerated in healthy volunteers, with the exception of some injection site erythema (ISE) in 3 out of 6 subjects, although without pain, induration, tenderness, or swelling. All subjects recovered spontaneously. No clinically significant changes were observed in vital signs, laboratory parameters (hematology, blood chemistry, coagulation, and urinalysis), physical examination, or ECG.

[0270] R5000 was measured in the 0.20 mg / kg dose group of the multi-dose arm of the study (see table below). In the table below, C max AUC refers to the highest plasma drug concentration. 0-24 This refers to the area under the plasma concentration-time curve from time 0 to 24 hours.

[0271] [Table 16]

[0272] Example 15. Phase 1 single-dose escalation clinical trial of R5000 A phase 1 single-dose escalation clinical pharmacology study was conducted in healthy human volunteers to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of R5000 after subcutaneous (SC) injection. The study was a randomized, double-blind, and placebo (PBO) controlled trial involving four SC single-dose escalation cohorts housed in a clinical pharmacology unit for three days. All subjects received one dose of R5000 on day 1. Four subjects (two receiving R5000 and two receiving PBO) received the lowest dose level (0.05 mg / kg), and six subjects per cohort (four receiving R5000 and two receiving PBO) received three higher dose levels (0.1, 0.2, and 0.4 mg / kg) sequentially. Demographic information for the subjects is presented in the table below.

[0273] [Table 17]

[0274] Safety was assessed through intensive clinical monitoring, and blood samples were frequently obtained to measure R5000 concentration by liquid chromatography / high-resolution mass spectrometry and its ability to inhibit complement-mediated RBC lysis in ex vivo antibody-sensitized sheep erythrocyte hemolysis assays.

[0275] The pharmacokinetic (PK) parameters measured in this study were clearance (CL) and C max (Maximum plasma drug concentration, Figure 11A), T max (the time it takes for a drug to reach its peak plasma concentration), t 1 / 2 (Half-life), AUC 0-24 (Area under the curve of plasma concentration over time from 0 to 24 hours; see Figure 11B for plasma concentration over time), AUC 0-inf (Area under the curve of plasma concentration versus time from time 0 to infinity; see Figure 11B for plasma concentration over time), V z This includes (apparent distribution volume in the terminal phase), K (efflux rate), and F (fraction). The results for each parameter are shown in the table below.

[0276] [Table 18]

[0277] All cohorts match the predictions from the in silico PK model created using data from non-human primate (NHP) studies. max The level was achieved. The plasma concentration of a single SC injection was C max A linear relationship was observed between the dose level and the mean peak plasma concentration (C). (Figure 11A) max The concentration ranged from 1010 to 5873 ng / mL throughout administration. The mean area under the concentration-time curve (AUC) from 0 to 24 hours post-administration was also measured. 0-24 ) administered to a dose of 21,440 to 112,300 ng * The range was time / mL. These results showed that plasma concentration (C) increased with increasing dose of R5000. max ) and exposure (AUC 0-24 This indicates an increase that is roughly proportional to the time to reach the peak observed plasma concentration (t). max The median absorption time was in the range of 3.0–4.6 hours throughout administration, indicating that R5000 exhibits an intermediate absorption rate from the SC space to the central (blood) compartment. The mean apparent systemic clearance (CL / F) after R5000 administration was low, ranging from 0.2481–0.4711 mL / hour / kg. Mean half-life (t 1 / 2 The apparent total volume of distribution (V) in the terminal phase after extravasation was consistent throughout the dose level and ranged from 155.6 to 185.4 hours. z / F) The mean range was 61.89–105.1 mL / kg, which indicates that R5000 was mainly localized within the circulating blood compartment and had minimal extravascular distribution. Approximate t across the entire cohort 1 / 2 It was determined to be 7 days.

[0278] R5000 also exhibits rapid dose-dependent inhibition of hemolysis [direct hemolysis (Figure 12A) and %CH]. 50(Figure 12B) and time-course erythrocyte lysis in 1% plasma (Figure 12C) were observed, along with inhibition of complement activity (measured by Wieslab® ELISA in all subjects after a single dose; see Figure 13). The maximum pharmacodynamic effect was observed approximately 3 hours after administration. The results showed that at the highest plasma concentration, the maximum percentage inhibition of hemolysis compared to baseline reached >90% in the 0.1, 0.2, and 0.4 mg / kg dose cohorts, and 60% in the lowest dose (0.05 mg / kg) cohort. Dose-dependent inhibition of hemolysis lasted up to 4 days in the 0.1, 0.2, and 0.4 mg / kg dose cohorts. In particular, mean hemolysis remained above baseline for up to 2 days in the 0.05 mg / kg cohort, up to 4 days in the 0.1 mg / kg cohort, and up to 7 days in the 0.2 and 0.4 mg / kg cohorts.

[0279] Similarly, complement activity analysis showed that the inhibition of complement activity remained potent for 4 days after a 0.4 mg / kg injection (see Figure 13). Human plasma samples obtained from subjects receiving a 0.4 mg / kg injection were subjected to Wieslab® ELISA (Euro Diagnostica, Malmö, Sweden). This assay measures alternative pathways for complement activity. When measured via this assay, complement activity was suppressed by 3% at 3 hours post-administration and remained below 13% 96 hours after R5000 administration.

[0280] A single dose of R5000 via SC was safe and well-tolerated in healthy volunteers. ISE (isotropic edema) was observed in three subjects at the highest dose, but it was mild (grade 1) without pain, induration, tenderness, or swelling, and resolved spontaneously within 2–5 hours post-injection. No clinically significant changes were observed in vital signs, laboratory parameters, physical examination, or ECG.

[0281] This study suggests that a steady-state level suitable for >80% suppression of hemolysis can be achieved with a low daily dose, and that once-weekly administration may be sufficient. Specifically, 0.2 mg / kg may result in complete suppression of complement activity and complete inhibition of hemolysis. (Note) As a preferred embodiment, the technical concept that can be understood from the above embodiment is described below. [Item 1] A pharmaceutical composition comprising R5000 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises sodium chloride at a concentration of about 25 mM to about 100 mM and sodium phosphate at a concentration of about 10 mM to about 100 mM. [Item 2] The pharmaceutical composition described in item 1, wherein R5000 is present at a concentration of approximately 1 mg / mL to approximately 400 mg / mL. [Item 3] A pharmaceutical composition as described in item 1, containing a pH of approximately 6.5 to approximately 7.5. [Item 4] R5000 has an equilibrium dissociation constant (K) of approximately 0.1 nM to approximately 1 nM. D A pharmaceutical composition as described in item 1, wherein the C5 is bonded at ) [Item 5] The pharmaceutical composition according to item 1, wherein R5000 blocks the production of C5a following activation of an alternative pathway for complement activation. [Item 6] The pharmaceutical composition described in item 1, wherein R5000 blocks the formation of membrane invasion complexes (MACs) following activation of the classical, alternative, or lectin pathways of complement activation. [Item 7] A method for inhibiting hemolysis in a subject, comprising administering a pharmaceutical composition described in any one of items 1 to 6. [Item 8] The method according to item 1, wherein the pharmaceutical composition is administered in a dose sufficient to achieve a plasma R5000 level of approximately 0.1 μg / mL to approximately 20 μg / mL. [Item 9] The method described in item 8, which inhibits hemolysis by approximately 25% to 100% after administration. [Item 10] The method according to item 9, wherein the pharmaceutical composition is administered daily for at least two days. [Item 11] The method according to item 9, wherein the pharmaceutical composition is administered daily for 7 days. [Item 12] The method according to item 9, wherein the pharmaceutical composition is administered daily for at least 100 days. [Item 13] The method described in any one of items 9 to 12, wherein no adverse effects on the cardiovascular, respiratory, and central nervous system (CNS) are observed for at least one month after administration. [Item 14] The method described in item 13, wherein no change is observed in at least one of the heart rate and arterial pressure for at least one month after administration. [Item 15] The method according to item 13, wherein no change is observed in at least one of the respiratory rate, tidal volume, and minute ventilation for at least one month after administration. [Item 16] The method according to any one of items 7 to 15, wherein the pharmaceutical composition is administered subcutaneously (SC) or intravenously (IV). [Item 17] Half-life (t) of R5000 levels in the target plasma 1 / 2 The method described in item 16, wherein the duration is at least 4 hours. [Item 18] t levels of R5000 in the target plasma 1 / 2 The method described in item 16, which takes approximately 1 to 10 days. [Item 19] The method described in item 16, wherein the steady-state distribution volume of R5000 in the target plasma is approximately 10 mL / kg to approximately 200 mL / kg. [Item 20] The method according to item 16, wherein the steady-state distribution volume of R5000 in the target plasma is equal to at least 50% of the total blood volume. [Item 21] The method described in item 16, wherein the total clearance rate of R5000 in the target plasma is approximately 0.04 mL / hour / kg to approximately 4 mL / hour / kg. [Item 22] T of R5000 in the target plasma max The method described in item 16, which is approximately 1 hour to approximately 48 hours. [Item 23] The method described in item 16, wherein the presence of a measurable amount of R5000 is substantially limited to the plasma compartment. [Item 24] The method according to item 7, wherein the pharmaceutical composition is administered in a dose sufficient to deliver R5000 in an amount of about 0.01 mg to about 2 mg / kg of body weight. [Item 25] The method described in item 24, which inhibits approximately 50% to 99% of C5 activation in the subject. [Item 26] The method according to item 7, wherein the pharmaceutical composition is administered in a dose sufficient to deliver R5000 at a dose of approximately 0.1 mg to approximately 0.4 mg / kg of target body weight. [Item 27] The method according to any one of items 24 to 26, wherein the pharmaceutical composition is administered subcutaneously or intravenously. [Item 28] The method according to item 27, wherein the pharmaceutical composition is administered once or more times daily. [Item 29] The method according to item 28, wherein the pharmaceutical composition is administered over a period of 7 days. [Item 30] The method described in item 27, wherein the percentage inhibition of hemolysis up to 3 hours after the first dose is at least 90% to about 95% or more. [Item 31] The method described in item 27, wherein the percentage inhibition of hemolysis is at least 90% to about 95% or more, as measured at least 7 days after administration. [Item 32] The method described in item 27, wherein the percentage inhibition of hemolysis is at least 90% to about 95% or more for at least 4 days after administration. [Item 33] The method according to item 27, wherein at least one of the maximum inhibition of hemolysis and the maximum inhibition of complement activity is achieved approximately 2 hours to approximately 4 hours after administration. [Item 34] The method described in item 33, in which R5000 is administered at a dose of 0.2 mg / kg. [Item 35] The method described in item 28, wherein hemolysis is 3% or less 24 hours after the final dose. [Item 36] The method described in item 29, wherein complement activity decreases to approximately 1 percent to approximately 10 percent during the aforementioned 7 days. [Item 37] The method described in item 28, wherein complement activity is 5% or less 24 hours after the final dose. [Item 38] The method according to item 7, wherein the pharmaceutical composition is administered daily by subcutaneous or intravenous injection in a dose sufficient to deliver R5000 in an amount of about 0.1 mg / day to about 60 mg / day / kg of target body weight. [Item 39] The highest serum concentration achieved (C max The method described in item 38, wherein the concentration is approximately 0.1 μg / mL to approximately 1000 μg / mL. [Item 40] Area under the curve (AUC) is approximately 200μg * time / mL ~ approx. 10,000μg * The method described in item 38, which is time / mL. [Item 41] A method for treating paroxysmal nocturnal hemoglobinuria (PNH) in a subject requiring such treatment, comprising subcutaneous or intravenous administration of a pharmaceutical composition described in any one of items 1 to 6. [Item 42] The method described in item 41, wherein the subject has been previously treated with an antibody-based therapeutic agent. [Item 43] The method according to item 42, wherein the PNH in the subject showed resistance or unresponsiveness to treatment using the antibody-based therapeutic agent. [Item 44] The method according to item 42 or 43, wherein the antibody-based therapeutic agent is eculizumab. [Item 45] A kit comprising a pharmaceutical composition described in any one of items 1 to 6 and instructions for its use. [Item 46] An automated injection device containing a pharmaceutical composition as described in any one of items 1 to 6. [Sequence List] [ka] [ka]

Claims

1. a) C5 inhibitors, b) Sodium chloride, and c) A water-soluble pharmaceutical composition comprising sodium phosphate, The C5 inhibitor has the following sequence: ______________________________________________P__________________ １ ︁ ２ ____________________________ ３ .......................... ４ yys Includes, Xaa １ However, it is N-methyl-aspartic acid, Xaa ２ However, it is tert-butylglycine, Xaa ３ However, it is 7-azatryptophan, Xaa ４ However, it is a water-soluble pharmaceutical composition that is cyclohexylglycine.

2. The pharmaceutical composition according to claim 1, wherein the C5 inhibitor comprises a lactam crosslink between the residues at position 1 and position 6.

3. The pharmaceutical composition according to claim 1 or 2, wherein the N-terminus of the C5 inhibitor is acetylated.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the C-terminal residue of the C5 inhibitor is lysine modified with a lysine side chain portion containing PEG and a saturated or unsaturated fatty acid.

5. The pharmaceutical composition according to claim 4, wherein a linker is present between the PEG and the fatty acid group.

6. A water-soluble pharmaceutical composition for use as a pharmaceutical, a) C5 inhibitors, b) Salts selected from the group consisting of acetates, chlorides, carbonates, and sulfates, c) Contains a buffering agent, The C5 inhibitor has the following sequence: ______________________________________________P__________________ １ ︁ ２ ____________________________ ３ .......................... ４ yys Includes, Xaa １ However, it is N-methyl-aspartic acid, Xaa ２ However, it is tert-butylglycine, Xaa ３ However, it is 7-azatryptophan, Xaa ４ However, it is cyclohexylglycine, A pharmaceutical composition administered subcutaneously.

7. The pharmaceutical composition according to claim 6, wherein the C5 inhibitor comprises a lactam crosslink between the residues at positions 1 and 6.

8. The pharmaceutical composition according to claim 6 or 7, wherein the N-terminus of the C5 inhibitor is acetylated.

9. The pharmaceutical composition according to any one of claims 6 to 8, wherein the C-terminal residue of the C5 inhibitor is lysine modified with a lysine side chain portion containing PEG and a saturated or unsaturated fatty acid.

10. The pharmaceutical composition according to claim 9, wherein a linker is present between the PEG and the fatty acid group.

11. The pharmaceutical composition according to any one of claims 6 to 10, wherein the salt is sodium chloride.

12. The pharmaceutical composition according to any one of claims 6 to 11, wherein the buffering agent comprises sodium phosphate.

13. The pharmaceutical composition according to any one of claims 6 to 12, wherein the pharmaceutical composition is self-administered.

14. The pharmaceutical composition according to any one of claims 6 to 13, wherein daily subcutaneous administration provides a steady-state inhibitor concentration that reaches within 1 to 3 doses, 2 to 3 doses, 3 to 5 doses, or 5 to 10 doses.

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