RU2779495C2 - Dosing schemes and corresponding compositions and methods - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: methods for the treatment of a subject, who needs the treatment of a complement-mediated disorder, are proposed, including the administration of a long-acting compstatin analogue (hereinafter – LACA) containing PEG with an average molecular weight of 40 kDa, subcutaneously at a dose of 1080 mg. A single dosed composition of LACA containing PEG with an average molecular weight of 40 kDa is proposed for the treatment of a complement-mediated disorder in a subject for subcutaneous administration in the amount of 1080 mg. A device is proposed for delivery of a drug containing the specified single dosed composition.

EFFECT: proposed group of inventions provides for increase in doses of a long-acting compstatin analogue containing 40 kDa of PEG.

27 cl, 28 dwg, 23 tbl, 30 ex

Description

CROSS REFERENCES TO RELATED APPLICATIONS

[0001] This application claims priority under 35 U.S.C. § 119(e) U.S.C. Provisional Applications No. 62/483,295 filed April 7, 2017 and U.S. Provisional Application No. 62/485,343 filed April 13, 2017, in full each of which is incorporated herein by reference in its entirety.

State of the art

[0002] The complement system is a system of more than 30 plasma proteins and cell-associated proteins that plays an important role in both innate and adaptive immunity. Complement proteins function as a series of enzymatic cascades through a variety of protein interactions and cleavage processes. Complement activation occurs through three main pathways: the antibody-dependent classical pathway, the alternative pathway, and the mannose-binding lectin (MBL) pathway. Inappropriate or excessive complement activation is a major cause or contributory factor in a number of serious diseases and conditions, and over the past few decades, significant efforts have been directed to the study of various complement inhibitors for use as therapeutic agents.

Short description

[0003] In some aspects, the present invention provides certain analogs of long-acting compstatin and/or the present invention relates to such analogs. For example, the present invention provides and/or the present invention relates to such long acting compstatin analogues, compositions containing them, and methods for their preparation, identification, characterization and/or use. In some aspects, the present invention provides a physiologically acceptable composition comprising a long acting compstatin analogue and/or the present invention relates to said composition. In some aspects, the present invention provides a pharmaceutical grade composition comprising a long acting compstatin analogue and/or the present invention relates to said composition. Among other things, in certain aspects, the present text describes particularly suitable long-acting compstatin analogues and, moreover, suggests specific doses, dosage formats, administration schedules, unit dosage compositions, and other technologies associated with administering long-acting compstatin analogues to human subjects, for example, specific human subjects, for example, suffering from one or more specific diseases, disorders or conditions and/or susceptible to said diseases, disorders or conditions.

[0004] In some aspects, the present invention provides methods for treating a subject in need of treatment for a complement-mediated disorder, which methods may include administering to said subject a long-acting compstatin analog using specific doses, dosage formats (e.g., unit dosage formulations, and/or specific formulations) and/or administration schedules (eg, routes of administration, dosage intervals, etc. in some embodiments that have been found to be particularly useful in the treatment of certain diseases, disorders, or conditions).

[0005] In some embodiments, the complement-mediated disorder being treated according to the present invention is paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), or another disorder associated with complement-mediated hemolysis. In some embodiments, said disorder is an ischemic/reperfusion (I/R) injury (eg, from myocardial infarction, thromboembolic stroke, or surgery). In some embodiments, said violation is an injury. In some embodiments, said violation is graft rejection. In some embodiments, said disorder is a chronic respiratory disorder, such as asthma or COPD (chronic obstructive pulmonary disease).

[0006] All articles, books, patent applications, patents, other publications, websites and databases referenced herein are incorporated by reference herein. In the event of a conflict between the present disclosure and any of the incorporated references, the present disclosure (including any amendments thereto) shall govern. Unless otherwise indicated, the meanings of terms and abbreviations accepted in the respective fields of technology are used in this document. Some of the aspects described herein may utilize standard techniques in molecular biology, cell culture, recombinant nucleic acid (e.g., DNA), immunology and/or synthesis, detection, manipulation, and quantification of nucleic acids and polypeptides, and the like. corresponding to the qualifications of a person skilled in the art. See, for example: Ausubel, F., et al., (eds.), Current Protocols in Molecular Biology, Current Protocols in Immunology, Current Protocols in Protein Science, Current Protocols in Cell Biology, published by John Wiley & Sons, N.Y. , for example, the edition current as of January 2010 or later; Sambrook, Russell, Sambrook, Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2001; 4th ed., 2012.

Brief description of the drawings

[0007] FIG. 1 is a graph showing the percent complement activation inhibitory activity of the compstatin analog CA28 (SEQ ID NO: 28) and three long acting compstatin analogs (CA28-1, CA28-2, CA28-3) as a function of peptide concentration (μM). Inhibition of complement activation was studied in vitro using the classical complement pathway inhibition assay. The graph shows the values obtained by averaging the results of two series of measurements. CA28 (circles; red), CA28-1 (crosses (x); blue); CA28-2 (triangles, green), CA28-3 (squares (purple)).

[0008] FIG. 2 is a graph showing the percent complement activation inhibitory activity of CA28 and the long acting compstatin analogs CA28-2 and CA28-3 as a function of compound concentration (µM). CA28 (squares, light gray), CA28-2 (diamonds, black), CA28-3 (circles, dark gray). CA28-3 is a compound that contains multiple peptide fragments (ie multiple fragments). Although the activity per peptide fragment is less than the activity of a single CA28 molecule, the total activity of CA28-3 in terms of moles is higher than the activity of CA28.

[0009] FIG. 3 is a plot of CA28 and long-acting compstatin analogues CA28-2 and CA28-3 in plasma concentrations versus time in cynomolgus monkeys following a single intravenous injection. CA28 was administered at 200 mg/kg. CA28-2 and CA28-3 were each administered at 50 mg/kg. When calculating the doses for these experiments, it was assumed that the administered substances CA28-2 and CA28-3 contain 80 wt. % active compound, based on dry weight. However, when analyzing samples, the standard curve implied 100 wt. % active compound, based on dry weight, by about 30%. Thus, the Cmax values are higher than the real Cmax. CA28 (squares, light grey), CA28-2 (triangles, black), CA28-3 (circles, dark grey).

[0010] FIG. 4 is a graph showing the percentage of complement activation inhibitory activity of CA28 and long acting compstatin analogs CA28-4 as a function of compound concentration (µM). Inhibition of complement activation was studied in vitro using the classical complement pathway inhibition assay. The graph shows the values obtained by averaging the results of four series of CA28-4 measurements. CA28 (squares, light gray), CA28-4 (crosses, black).

[0011] In FIG. 5 is a graph showing concentrations versus time for CA28 and long-acting compstatin analogs CA28-2, CA28-3, and CA28-4 in cynomolgus monkeys after a single intravenous injection. CA28 was administered at 200 mg/kg. Each of CA28-2, CA28-3 and CA28-4 was administered in an amount of 50 mg/kg. When calculating the doses for these experiments, it was assumed that the administered substances CA28-2 and CA28-3 contain 80 wt. % active compound, based on dry weight. However, when analyzing samples, the standard curve implied 100 wt. % active compound, based on dry weight. Thus, the Cmax values exceed the Cmax values that would be obtained with the introduction of these compounds in the indicated doses, calculated on a dry weight, by about 30%. CA28 (squares, light grey), CA28-2 (triangles, black), CA28-3 (circles, dark grey), CA28-4 (inverted triangles, black).

[0012] FIG. 6 is a chromatogram showing ultraviolet (UV) detection of a long-acting PEG-based compstatin analogue using reverse phase HPLC. The peak with a retention time (RT) of 33.68 minutes corresponds to the pegylated analog of compstatin and has a relative area of 96%.

[0013] FIG. 7 is a graph showing the percent complement activation inhibitory activity of CA28 and the long acting compstatin analogs CA28-2CS, CA28-2GS, CA28-2HS and CA28-2TS as a function of compound concentration (µM). CA28-2CS (diamonds, red); CA28-2GS (crosses, blue); CA28-2HS (triangles, green); CA28-2TS (squares, black).

[0014] FIG. 8 is a graph showing the percentage of complement activation inhibitory activity of CA28 and the long-acting bifunctionalized compstatin analog, CA28-2GS-BF, as a function of compound concentration (micromoles). CA28 (open circles, blue); CA28-2G-SBF (solid circles, red)

[0015] FIG. 9 is a graph showing the plasma concentration versus time of CA28 and the long-acting compstatin analog CA28-2GS-BF in cynomolgus monkeys after either a single intravenous injection (CA28 (squares, red) and CA28-2GS-BF (circles, purple), or administered by subcutaneous injection once a day for 7 days (only CA28-2GS-BF, asterisks, blue) CA28-2GSBF was administered at a concentration of 25 mg/ml Dose volume was 2 ml/kg for iv (intravenous) and 0.28 ml/kg/day for subcutaneous administration.Data for CA28 were obtained in another experiment in which the indicated compound was also contained in 5% dextrose, and its concentration was 20 mg/ml at a dose volume of 10 ml /kg The basis in each case was represented by 5% dextrose in water.

[0016] FIG. 10(A) and 10(B) are graphs showing the percentage of complement activation inhibitory activity of CA28 and the long-acting bifunctionalized compstatin analog, CA28-2TS-BF, as a function of compound concentration (micromoles). (A) Classical pathway inhibition by CA28 (circles, red) and CA28-2TS-BF (crosses, blue). (B) Inhibition of the alternative pathway. CA28 (circles, red) and CA28-2TS-BF (crosses, blue).

[0017] FIG. 10(C) (at 40 kDa PEG fragment size) shows the structure of CA28-2TS-BF.

[0018] FIG. 11 is a graph showing the plasma concentration versus time of CA28 and the long-acting compstatin analog CA28-2TS-BF in cynomolgus monkeys after a single intravenous injection of CA28 at a dose of 200 μg/kg (squares, red), a single intravenous injection of CA28-2TS- BF at 7 mcg/kg (asterisks, purple), a single subcutaneous injection of CA28-2TS-BF at a dose of 7 mcg/kg (circles, blue) or a subcutaneous injection of CA28-2TS-BF at a dose of 7 mcg/kg once a day day for 7 consecutive days (inverted triangles, green). The base in each case was 5% dextrose in water.

[0019] FIG. 12 shows flow cytometry analysis of C3 deposition on red blood cells from a PNH patient exposed to activated complement in a modified Ham test. (A) Shown are the results of a dilution experiment demonstrating the effect of CA28 on C3 deposition. (B) Results of a dilution experiment showing the effect of CA28-2GS-BF on C3 deposition are shown. The compound concentrations used are shown on and above each panel.

[0020] FIG. 13 shows flow cytometry analysis of C3 deposition on red blood cells from a PNH patient exposed to activated complement in the modified Ham test in the absence of complement inhibitors (left panel), in the presence of the anti-C5 monoclonal antibody eculizumab (middle panel), and in the presence of CA28. -2GS-BF (right panel).

[0021] In FIG. 14 is a graph of ex vivo serum-induced hemolysis observed in a multiple escalating dose study of a long acting compstatin analog containing 40 kDa PEG in healthy subjects.

[0022] FIG. 15 is a schematic diagram of a target-mediated drag disposition (TMDD) model.

[0023] FIG. 16 shows the observed and predicted serum concentrations of LACA-40 during the nominal time after administration, divided by dosing regimen, for the single dose (s.d.) and multiple dose study over 28 days (ppm, once per day) in healthy subjects. The solid circles represent the observed mean PK (pharmacokinetic) concentrations at the respective nominal time points overlaid with the TMDD model predictions represented by the solid line. Each administration scheme is represented by a different color. LACA-40 was administered subcutaneously.

[0024] FIG. 17 shows the predicted residual (minimum) serum PK concentrations during the nominal time after administration after repeated daily (q.d.) subcutaneous (s.c.) administration of LACA-40 at three dose levels: 180, 270, and 360 mg q.d. . The predictions were based on the TMDD model.

[0025] In FIG. 18 shows the predicted residual serum PK concentrations over a nominal time after dosing after multiple daily (qd) s.c. and s.c. three times weekly (Monday/Wednesday/Friday) doses of LACA-40. Each injection scheme is represented by a distinct color line and the predictions were based on the TMDD model.

[0026] FIG. 19 shows the predicted residual serum PK concentrations over a nominal time after administration after multiple daily (qd) s.c. and s.c. twice a week (Monday/Thursday) doses of LACA-40. Each injection scheme is represented by a distinct color line and the predictions were based on the TMDD model.

[0027] In FIG. 20 shows the predicted residual serum PK concentrations over a nominal time after administration after multiple daily (qd) s.c. and once a week s.c./w. doses of various doses of LACA-40. Each injection scheme is represented by a distinct color line and the predictions were based on the TMDD model.

[0028] FIG. 21 shows the predicted residual serum PK concentrations over a nominal time after administration after multiple daily (qd) s.c. and once a week s.c./w. doses of various doses of LACA-40. Each injection scheme is represented by a distinct color line and the predictions were based on the TMDD model.

[0029] FIG. 22 shows the predicted serum concentrations during the nominal time after administration after multiple daily (q.d.) s.c., s.c. /n) of different doses of LACA-40 over a 28 day administration period corresponding to the administration schedules for cohorts 1, 2 and 3 in the clinical study described in Example 30. Each administration regimen is represented by a distinct color line and predictions were based on the TMDD model . For the twice-weekly schedule, the schedule reflects administration at alternating intervals of days 3 and 4 (ie, administration on days 1, 4, 8, 11, 15, 18, 22, and 25).

[0030] FIG. 23 shows an expanded view of the predictions up to day 35 for the LACA-40 concentrations shown in FIG. 22 for daily administration and administration twice a week for a period of 28 days.

[0031] In FIG. 24 shows plots of total observed individual and mean serum LACA-40 concentrations for cohorts 1 and 2 in the clinical study described in Example 30.

[0032] FIG. 25 shows plots of the total observed individual and mean serum concentrations of LACA-40 for cohort 1 in the clinical study described in Example 30 and the serum concentrations for this regimen predicted based on the TMDD model.

[0033] FIG. 26 shows graphs of the total observed individual and mean serum concentrations of LACA-40 for cohort 2 in the clinical study described in Example 30 and the serum concentrations for this regimen predicted based on the TMDD model.

[0034] FIG. 27 shows plots of mean total observed serum concentrations of LACA-40 for cohorts 1 and 2 in the clinical study described in Example 30 and serum concentrations for these dosing schedules predicted based on the TMDD model. The plots further illustrate that the observed FK data are consistent with the predictions of the TMDD model.

[0035] FIG. 28 shows the mean total observed serum concentrations of LACA-40 for cohorts 1 and 2 in the clinical study described in Example 30 and the serum concentrations for cohort 3 predicted based on the TMDD model.

Detailed Description of Some Implementations

[0036] I. Definitions

[0037] The terms "about"' or "about" in relation to a number typically include numbers falling within the range of ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0, 5% of the number, unless otherwise specified or otherwise evident from the context (unless such number unacceptably exceeds 100% of the possible value).

[0038] A "complement component" or "complement protein" is a protein involved in the activation of the complement system or involved in one or more complement-mediated activities. Components of the classical complement pathway include, for example, the C1q, C1r, C1s, C2, C3, C4, C5, C6, C7, C8, C9, and C5b-9 complex, also called the membrane attack complex (MAC), and active fragments or products of the enzyme cleavage of any of the above components (for example, C3a, C3b, C4a, C4b, C5a, etc.). Alternative pathway components include, for example, factors B, D, and properdin. Components of the lectin pathway include, for example, MBL2, MASP-1 and MASP-2. Complement components also include cell-bound receptors for soluble complement components, characterized in that said receptor mediates one or more biological activities of such a soluble complement component after binding of said soluble complement component. Such receptors include, for example, the C5a receptor (C5aR), the C3a receptor (C3aR), the complement receptor 1 (CR ¹ ), the complement receptor 2 (CR2), the complement receptor 3 (CR3, also known as CD45), and the like. It should be understood that the term "complement component" is not intended to include those molecules and molecular structures that serve as "triggers" for complement activation, such as antigen-antibody complexes, foreign structures found on microbial or artificial surfaces, and etc.

[0039] A "complement-mediated disorder" is any disorder for which, in at least some subjects suffering from said disorder, complement activation is known to be a contributing and/or at least partly causative factor; for example, disorders in which complement activation leads to tissue damage. Non-limiting examples of complement-mediated disorders include, but are not limited to, (i) various disorders characterized by hemolysis or hemolytic anemia such as atypical hemolytic uremic syndrome, autoimmune hemolytic anemia with warm agglutinin syndrome, cold agglutinin disease, paroxysmal nocturnal hemoglobinuria, transfusion reactions ; (ii) graft rejection (eg, hyperacute or acute graft rejection) or graft dysfunction; (iii) disorders involving ischemic/reperfusion injury such as trauma, surgery (eg, aneurysm repair), myocardial infarction, ischemic stroke; (iv) respiratory disorders such as asthma and chronic obstructive pulmonary disease (COPD); (v) arthritis, eg rheumatoid arthritis; (vi) ocular disorders such as age-related macular degeneration (AMD), diabetic retinopathy, glaucoma and uveitis. The term "disorder" is used herein interchangeably with the terms "disease", "condition" and similar terms, to refer to any disorder of health or condition of abnormal functioning of the body, for example, any condition in which the organization of medical care and / or surgical intervention is indicated, or in connection with which the subject seeks appropriate medical and/or surgical care. It should also be understood that specific violations are listed in specific categories for convenience and this is not intended to limit the present invention. It should be understood that certain violations may be listed in several suitable categories.

[0040] "Complement regulatory protein" (CRP) is a protein involved in the regulation of complement activity. The complement regulatory protein can down-regulate complement activity, for example by inhibiting complement activation or by inactivating or accelerating the destruction of one or more activated complement proteins. Examples of complement regulatory proteins include C1 inhibitor, C4 binding protein, clusterin, vitronectin, CFH, factor I, and cell-associated proteins CD46, CD55, CD59, CR ¹ , CR2 and CR3.

[0041] "Separated" as used herein means 1) separated from at least some of the components with which there is normally an initial association in nature; 2) processed or purified using a process involving human intervention; and/or 3) not naturally occurring, eg, present in the built environment. In general, unless otherwise indicated or otherwise evident, any substance, product, agent, composition, and the like, may be considered "isolated" if necessary.

[0042] As used herein, "linked" in relation to two or more fragments means that said fragments are physically linked or connected to each other to form a sufficiently stable molecular structure such that said fragments remain linked under the conditions under which said bond is formed, and preferably under conditions under which said novel molecular structure is used, eg under physiological conditions. According to certain preferred embodiments of the present invention, said bond is a covalent bond. In other embodiments, said bond is non-covalent. Fragments can be linked either directly or indirectly. If two fragments are directly linked, they are either covalently linked to each other or are at a sufficiently close distance so that their bond is maintained due to intermolecular forces between these two fragments. If two fragments are indirectly linked, each of them is connected either covalently or non-covalently with a third fragment, due to which the connection between the two fragments is maintained. In general, when two moieties are said to be linked by a "linking moiety" or "linking part", the bond between said two linked moieties is indirect, and typically each of the linked moieties is covalently bonded to the linking moiety. Two fragments can be connected using a "linker". The linker may be any suitable moiety that reacts with said compounds to bind them in a reasonable amount of time, under conditions appropriate to the stability of said compounds (the portions of which may be protected as appropriate, depending on the conditions), in sufficient quantity, with a satisfactory outcome. Typically, the linker contains at least two functional groups, one of which reacts with the first connection, and the other reacts with the second connection. It should be understood that after the linker has reacted with the compounds to be linked, the term "linker" may refer to the part of the resulting structure that is derived from said linker, or at least the part that does not include the reacted functional groups. The connecting fragment may contain a part that is not involved in the formation of bonds with the connected compounds, the main purpose of which may be the spatial separation of these compounds. Such a part may be called a "spacer".

[0043] As used herein, the phrase "physiological conditions" refers to a set of conditions, such as temperature, salt concentration, pH, that at least partially mimic conditions typically found in a living subject, e.g., a mammalian subject. In some aspects, physiological conditions refer to conditions in an aquatic environment, such as an environment containing at least 90%, 95%, 96%, 97%, 97%, 99%, or about 100% water by volume. In some embodiments, other fluids, if present, do not substantially affect the secondary or tertiary structure of the protein. In some embodiments, the physiological conditions at least partially mimic those found in a physiological fluid, such as blood or extracellular fluid, such as interstitial fluid, such as a mammalian subject. A variety of physiological conditions suitable for, for example, in vitro assays are known in the art. Typically, the environment under physiological conditions contains a physiological concentration of salt, such as sodium chloride. In some embodiments, physiological salt concentration refers to a concentration ranging from about 250 mOsm/L to about 350 mOsm/L, for example, from about 275 mOsm/L to about 325 mOsm/L, for example, about 300 mOsm/L. In some embodiments, physiological conditions are approximately isotonic with respect to bodily fluid, such as blood or extracellular fluid, such as interstitial fluid. In some embodiments, physiological conditions include pH values ranging from about 6.5 to about 7.8, such as from about 7.0 to about 7.5. In some embodiments, the physiological medium contains a buffering agent to help maintain the pH of the medium in the physiological range. In some embodiments, physiological conditions include conditions such that a typical mammalian protein, e.g., a protein that is typically present in a bodily fluid such as blood or extracellular fluid, substantially retains a secondary and, where appropriate, tertiary structure, which such a protein is present in the physiological fluid in which it is usually present. In some embodiments, components of the physiological medium are generally substantially non-toxic to mammalian cells at the concentration at which they are present in the physiological medium. A variety of physiological media (sometimes referred to as "buffers") are listed in various standard sources such as those cited above (eg, Sambrook, et al., Protocols series). In some embodiments, the physiological temperature ranges from about 25 degrees C to about 38 degrees C, for example, from about 30 degrees C to about 37 degrees C, for example, from 35 degrees C to 37 degrees C.

[0044] "Polypeptide" as used herein refers to an amino acid polymer optionally containing one or more amino acid analogs. A protein is a molecule made up of one or more polypeptides. The peptide is a relatively short polypeptide, typically about 2 to 60 amino acids in length, such as 8 to 40 amino acids in length. The terms "protein", "polypeptide" and "peptide" may be used interchangeably. The polypeptides herein may include amino acids such as those naturally occurring in proteins, amino acids not naturally occurring in proteins, and/or analogs of amino acids that are not amino acids. As used herein, an "analogue" of an amino acid may be another amino acid that is structurally similar to the specified amino acid, or a non-amino acid compound that is structurally similar to the specified amino acid. There are a significant number of analogs of the 20 amino acids commonly found in proteins ("standard" amino acids) known in the art. One or more amino acids of the polypeptide may be modified, for example, by adding a chemical compound such as a carbohydrate group, a phosphate group, a farnesyl group, an isofarnesyl group, a fatty acid group, a linker for conjugation, functionalization, or other modification, and the like. Certain suitable non-limiting analogs and modifications are described in WO 2004026328 and/or below. Said polypeptide may be acetylated, eg at the N-terminus and/or amidated, eg at the C-terminus.

[0045] The term "purified" as used herein refers to substances that have been separated from at least some or most of the components to which they are naturally or originally associated upon receipt, or to which they were associated prior to purification. In general, such cleansing involves human participation. Purified agents may be partially purified, substantially purified, or pure. Such agents may be, for example, at least 50% pure, 60% pure, 70% pure, 75% pure, 80% pure, 85% pure, 90% pure, 95% pure, 96% pure, 97% pure, 98% pure, 99% pure, or more than by 99%. In some embodiments, the nucleic acid, polypeptide or small molecule is purified such that it(a) is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more of the total amount of nucleic acid, polypeptide or small molecule material, respectively, present in the formulation. In some embodiments, an organic substance, such as a nucleic acid, polypeptide, or small molecule, is purified such that it is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more of the total amount of organic material present in the composition. Evaluation of purity can be based on, for example, dry weight, peak size in chromatographic determination (GC, HPLC, etc.), abundance of molecules, electrophoretic methods, intensity of bands on the gel, spectroscopic data (for example, NMR), elemental analysis, high throughput sequencing, mass spectrometry, or any method of quantification accepted in the art. In some embodiments, water, buffers, ions, and/or small molecules (eg, synthetic precursors such as nucleotides or amino acids) may optionally be present in the purified formulation. The purified agent can be obtained by separation from other substances (eg, other cellular materials), or by synthesis in such a way that the required degree of purity is achieved. In some embodiments, "partially purified" in relation to a molecule produced by a cell means that the molecule produced by the cell is no longer present in said cell, e.g., the cell has been lysed and, optionally, at least some cellular material (e.g., cell wall , cell(s) membrane(s), cell(s) organelle(s) has been removed, and/or said molecule has been separated or isolated from at least some molecules of the same type (protein, RNA, DNA, etc. ) present in the lysate.

[0046] "Recombinant host cells", "host cells" and other similar terms refer to prokaryotic or eukaryotic cells or cell lines that contain exogenous nucleic acid (typically DNA), such as an expression vector containing a nucleic acid that encodes a polypeptide of interest. It should be understood that such terms include the descendants of the original(s) cell(s), which(s) was(a) introduced(a) vector or other nucleic acid. Suitable host cells include any cells commonly used in the art to express polynucleotides (e.g., to obtain the polypeptide(s) encoded by such polynucleotides), including, for example, prokaryotes such as E. coli or other bacteria, such as Escherichia species; Lactobacillus, Bacillus (eg B. subtilis), Salmonella, Pseudomonas, Streptomyces, Staphylococcus and the like; and eukaryotes including, for example, fungi such as yeasts (eg Pichia (eg Pichia pastoris), Kluyveromyces eg K. lactis, Hansenula eg H. polymorpha). Examples of other fungal cells are filamentous fungal cells, eg Aspergillus spp., Neurospora spp., Fusarium spp. or Trichoderma spp., eg strains of A. oryzae, A. nidulans or A. niger; insect cells (eg Sf9), plant and animal cells, eg mammalian cells such as CHO, R ¹ .1, BW, LM, African green monkey kidney cells (eg COS-1, COS-7, BSC- 1, BSC-40 and BMT-10) and cultured human cells. Also covered are genetically modified cells of genetically modified (eg, transgenic) plants and animals, characterized in that the recombinant polypeptide is produced by at least some of such cells. The polypeptide may be secreted into milk, obtained from plant material, and the like. The exogenous nucleic acid may be stably maintained as an episome, such as a plasmid, or may be at least partially integrated into the genome of the host cell, optionally after copying or reverse transcribing. Terms such as "host cells" and the like are also used to refer to cells or cell lines that can be used as recipients of exogenous nucleic acid prior to administration of the nucleic acid. A "recombinant polynucleotide" is generally a polynucleotide that contains nucleic acid sequences that do not naturally occur in a directly linked form. For example, said nucleic acid sequences may occur in different genes or in different species, or one or more of said sequences may be a variant of a naturally occurring sequence, or may at least partially be an artificial sequence that is not homologous to a naturally occurring sequence. A "recombinant polypeptide" is usually a polypeptide at least partially obtained by transcription and translation of an exogenous nucleic acid by a recombinant host cell or in vitro cell-free expression system and/or containing amino acid sequences that do not occur naturally in a directly linked form. In the latter case, the recombinant polypeptide may be referred to as a "chimeric polypeptide". Amino acid sequences in a chimeric polypeptide may, for example, occur in different genes or in different species, or one or more of the sequences may be a variant of a naturally occurring sequence, or may at least partially be an artificial sequence that is not identical, or in some embodiments which is not homologous to a naturally occurring sequence in a region constituting a significant portion of the length. It should be understood that a chimeric polypeptide may contain two or more polypeptides. For example, the first and second polypeptides A and B of the chimeric polypeptide may be directly linked (A-B or B-A) or may be separated by part of the third polypeptide C (A-C-B or B-C-A). In some embodiments, part C is a polypeptide linker, which may, for example, contain several glycine and/or serine residues, or any of a wide variety of other amino acids. In some embodiments, two or more polypeptides may be linked by non-polypeptide(s) linker(s). "'Recombinant" as used herein encompasses, in some embodiments, polypeptides obtained by the addition (eg, chemical conjugation, enzymatic conjugation) of shorter recombinant polypeptides that can be produced in recombinant host cells. In some embodiments, the recombinant polypeptide may contain a signal sequence directing the secretion of said polypeptide, or a sequence directing the expressed polypeptide to a particular compartment or organelle. Suitable sequences are known in the art. Sequences appropriate for the host cell type of interest (eg, bacterial, fungal, mammalian, plant, etc.) can be selected. In some embodiments, the signal sequence may be located at or near the N-terminus or C-terminus (eg, within 10-50 amino acids). In some embodiments, the polypeptide contains a label. The label may be useful for facilitating detection and/or purification of the protein containing it. Examples of tags include a polyhistidine tag (e.g. 6X-His tag) (SEQ ID NO:69), glutathione-S-transferase, maltose binding protein, NUS tag, SNUT tag, Strep tag, epitope tags such as V5, HA, Mus or FLAG. In some embodiments, the protease cleavage site is located in the region between the tag and the polypeptide, which allows separation of the polypeptide from the tag by exposure to the protease. In some embodiments, a polynucleotide that encodes a recombinant polypeptide is at least partially codon-optimized for expression in a host cell of interest (eg, bacterial, fungal, mammalian, plant, etc.). The tag can be located at or near the N- or C-terminus (eg, within 10-50 amino acids) of the polypeptide according to various embodiments. The recombinant polypeptide may be isolated, purified, and the like. using any of a variety of methods. See, for example, Sambrook, Protocols series, or other standard sources. Methods of use may include, for example, dialysis (for example, using membranes with a given pore size), chromatography, precipitation, gel purification, or affinity methods, in which, in some embodiments, a label or a specific binding reagent, such as an antibody, can be used.

[0047] The term "reactive functional groups" as used herein refers to groups including, but not limited to, olefins, acetylenes, alcohols, phenols, ethers, oxides, halides, aldehydes, ketones, carboxylic acids, esters, amides, cyanates , isocyanates, thiocyanates, isothiocyanates, amines, hydrazines, hydrazones, hydrazides, diazo groups, diazonium, nitro groups, nitriles, mercaptans, sulfides, disulfides, sulfoxides, sulfones, sulfonic acids, sulfonic acids, acetals, ketals, anhydrides, sulfates, isonitriles of sulfonic acid , amidines, imides, imidates, nitrones, hydroxylamines, oximes, hydroxamic acids, thiohydroxamic acids, allenes, orthoesters, sulfites, enamines, inamines, ureas, pseudoureas, semicarbazides, carbodiimides, carbamates, imines, azides, azo compounds, azoxy compounds and nitroso compounds, complex n-hydroxysuccinimide esters, maleimides, sulfhydryls, and the like. Methods for obtaining each of these functional groups are well known in the art, and their use or modification for a particular purpose is within the skill of the art (see, for example, Sandler and Karo, eds. ORGANIC FUNCTIONAL GROUP PREPARATIONS, Academic Press, San Diego, 1989, and Hermanson, G., Bioconjugate Techniques, 2nd ed., Academic Press, San Diego, 2008.

[0048] "Specific binding" generally refers to a physical bond between a target polypeptide (or, more generally, a target molecule) and a binding molecule, such as an antibody or ligand. Binding generally depends on the presence of a particular structural feature of the target, such as an antigenic determinant, epitope, binding pocket or recess, recognized by the binding molecule. For example, if an antibody is specific for an A epitope, the presence of a polypeptide containing an A epitope, or the presence of free unlabeled A in a reaction involving both free labeled A and a binding molecule attached to it, will reduce the amount of labeled A that binds to said binding molecule. . It should be understood that specificity need not be absolute, and generally refers to the context in which the binding occurs. For example, it is well known in the art that many antibodies cross-react with epitopes other than those present in the target molecule. Such cross-reactivity may be acceptable, depending on the intended use of the antibody. One of skill in the art will be able to select antibodies or ligands with a sufficient degree of specificity suitable for any particular application (eg, target molecule detection, therapeutic purposes, etc.). It should also be understood that specificity may be assessed in the context of additional factors such as the affinity of a binding molecule for a target relative to the affinity of said binding molecule for other targets, such as competitive ones. If a binding molecule exhibits high affinity for the target molecule to be detected with it and low affinity for non-target molecules, the antibody is likely to be an acceptable reagent. Once the specificity of a binding molecule in one or more contexts has been established, it can be used in other, preferably similar, contexts without the need to retest its specificity. In some embodiments, the affinity (as measured by the equilibrium dissociation constant, Kd) of the two molecules exhibiting specific binding is 10 ^-3 M or less, such as 10 ^-4 M or less, such as 10 ^-5 M or less, such as 10 ^-6 M or less, 10 ^-7 M or less, 10 ^-8 M or less, or 10 ^-9 M or less under the conditions tested, eg under physiological conditions.

[0049] A "subject" to be treated in accordance with the present invention is typically a human, non-human primate, or lower animal (e.g., mouse or rat) that expresses or has at least one of the - or components) of C3 complement of a primate (eg, human) and, optionally, one or more additional components of a primate complement. In some embodiments, said subject is a male. In some embodiments, said subject is a female. In some embodiments, said subject is an adult, such as a human at least 18 years of age, such as between 18 and 100 years of age. In some embodiments, the human subject is at least 12 years old. In some embodiments, the subject is an adult, such as a human at least 18 years of age, such as between 18 and 100 years of age. In some embodiments, the subject is at least 40, 45, 50, 55, 60, 65, 70, 75, or 80 years old. In some embodiments, said subject is a child, such as a human between 0 and 4 years of age, or between 5 and 11 years of age.

[0050] "Treatment" as used herein in relation to treating a subject refers to providing treatment, i. providing any type of organization of medical care or surgical treatment of the subject. Said treatment may be provided to reverse, alleviate, suppress the progression of, prevent or reduce the occurrence of a disease, or to reverse, alleviate, inhibit or prevent the progression of, prevent or reduce the occurrence of one or more symptoms or manifestations of a disease. "Prevent" refers to preventing the onset of a disease, the onset of a symptom, or the manifestation of a disease for at least some period of time in at least some individuals. Treatment may include administering a compound or composition to said subject after the development of one or more symptoms or manifestations indicative of a disease, for example, in order to reverse, alleviate, reduce the severity and/or suppress or prevent the progression of said disease and/or reverse, alleviate, reduce the severity and/or suppress one or more symptoms or manifestations of said disease. The compound or composition may be administered to a subject who has developed a disease or has an increased risk of developing said disease compared to a member of the general population. The compound or composition may be administered to a subject who has developed a disease and has an increased risk of developing one or more specific symptoms, manifestations of said disease, or exacerbations of said disease compared to other individuals diagnosed with said disease, or compared to typical or average risk. occurrence of such a symptom, manifestation or exacerbation in said subject. For example, said subject could be exposed to a “trigger” that causes an increased risk (in particular, a temporarily increased risk) of developing an exacerbation in said subject. The compound or composition may be administered prophylactically, i. before the development of any symptom or manifestation of said disease. As a rule, in this case, the subject has a risk of developing the specified disease, for example, relative to a member of the general population, not necessarily having the same age, sex and/or other demographic(s) indicator(s).

[0051] A "vector" can be any of a variety of nucleic acid molecules, viruses, or parts thereof capable of mediating entry, such as transfer, transport, and the like. nucleic acid of interest between different genetic environments or into a cell. The nucleic acid of interest can be linked, eg introduced into a vector using, eg, restriction and ligation. Vectors include, for example, DNA or RNA plasmids, cosmids, naturally occurring or modified viral genomes or portions thereof, nucleic acids that can be packaged into viral capsids, minichromosomes, artificial chromosomes, and the like. Plasmid vectors typically include an origin of replication (eg, for replication in prokaryotic cells). The plasmid may include part or all of the viral genome (e.g., viral promoter, enhancer, processing or packaging signals, and/or sequences sufficient to produce a nucleic acid that can be integrated into the host cell's genome and/or can cause the production of an infectious virus) . Viruses or parts thereof that can be used to introduce nucleic acids into cells may be referred to as viral vectors. Viral vectors include, for example, adenoviruses, adeno-associated viruses, retroviruses (for example, lentiviruses, vaccinia and other poxviruses, herpesviruses (for example, herpes simplex virus), etc. Baculovirus is suitable, for example, for use in insect cells. A significant number of plant viral vectors that include, for example, based on or containing cauliflower mosaic virus, tobacco mosaic virus, or one or more of their genetic elements (for example, the 35S promoter of cauliflower mosaic virus).Viral vectors may or may not contain sufficient viral genetic information to produce infectious virus when introduced into host cells, i.e. viral vectors may be replication competent or replication defective In some embodiments, for example, if sufficient information is not available to produce an infectious virus, it may be provided by the host cell or another vector , introduced into the specified cell, for example, is required to obtain a virus. In some embodiments, such information is not provided, for example, if obtaining a virus is not required. The transfer nucleic acid may be incorporated into a naturally occurring or modified viral genome or portion thereof, or may be present in the viral capsid as a single nucleic acid molecule. The vector may contain one or more nucleic acids encoding a marker suitable for identifying and/or selecting cells that have received the vector. Markers include, for example, various proteins that increase or decrease either resistance or sensitivity to antibiotics or other agents (for example, a protein conferring resistance to an antibiotic such as puromycin, hygromycin or blasticidin), enzymes whose activity can be detected using assays known in the art (e.g. (3-galactosidase or alkaline phosphatase), and proteins or RNAs that detectably affect the phenotype of cells expressing them (e.g., fluorescent proteins). Vectors often include one or more appropriately located sites for restriction enzymes that can be used to facilitate the insertion into a vector of a nucleic acid, for example, a nucleic acid to be expressed An expression vector is a vector into which the desired nucleic acid has been or can be inserted in such a way that it will be operably linked to regulatory elements nts (also called "regulatory sequences", "expression control elements", or "expression control sequences") and can be expressed as an RNA transcript (eg, mRNA that can be translated into a protein or non-coding RNA). Expression vectors include regulatory sequence(s), for example, expression control sequences, sufficient(s) to direct transcription of the operably linked nucleic acid under at least some conditions; other elements necessary for or facilitating expression may be provided, for example, by the host cell or in vitro expression system. Such regulatory sequences typically include a promoter and may include enhancer sequences or upstream activating sequences. In some embodiments, the vector may include sequences encoding a 5' untranslated region and/or a 3' untranslated region, which may contain a cleavage and/or polyadenylation signal. In general, the regulatory elements may be present in the vector prior to insertion of the nucleic acid to be expressed, or may be present in the inserted nucleic acid, or may be inserted into the vector after the nucleic acid to be expressed is inserted. As used herein, a nucleic acid and a regulatory element(s) are "operably linked" if they are covalently linked in such a way that expression or transcription of said nucleic acid is under the influence or control of said regulatory element(s). (s). For example, a promoter region is operably linked to a nucleic acid if said promoter region is capable of transcribing said nucleic acid. The person skilled in the art will recognize that the specific characteristics of regulatory sequences suitable for gene expression may vary between species or cell types, but may generally include as appropriate sequences involved in transcription initiation, RNA processing, or translation initiation. The selection and design of the appropriate vector and control element(s) is within the ability and skill of the skilled artisan. For example, one skilled in the art can select an appropriate promoter (or other expression control sequences) for expression in the desired cell species (e.g., prokaryotic (bacterial) or eukaryotic (e.g., fungal, plant, mammalian cells)) or type. The vector may contain a promoter capable of directing expression in mammalian cells, such as a suitable viral promoter, e.g., from cytomegalovirus (CMV), retrovirus, simian virus (e.g., SV40), papillomavirus, herpes virus, or other virus that infects mammalian cells, or a promoter mammalian, for example, from a gene such as EF1-alpha, a gene for ubiquitin (for example, ubiquitin B or C), globin, actin, phosphoglycerate kinase (PGK) and the like, or a composite promoter such as a CAG protomore (combination of early enhancer CMV element and chicken beta-actin promoter). In some embodiments, a human promoter may be used. In some embodiments, for example, a promoter that typically directs transcription by a eukaryotic RNA polymerase I ("pol I promoter") (U6, H1, 7SK or tRNA promoter, or a functional variant thereof) can be used. In some embodiments, a promoter is used that typically directs transcription by a eukaryotic RNA polymerase II (pol II promoter) or a functional variant thereof. In some embodiments, a promoter is used that typically directs transcription by a eukaryotic RNA polymerase III ("pol III promoter") , for example, a promoter for transcription of ribosomal RNA (non-5S rRNA) or a functional variant thereof. One of skill in the art can select an appropriate promoter to direct transcription of the sequence of interest. Examples of expression vectors that can be used in mammalian cells include, for example, pcDNA vector group, pSV2 vector group, pCMV vector group, pRSV vector group, pEF1 vector group, Gateway® vectors, and the like. In some embodiments, regulable (e.g., inducible(s) or repressible(s)) expression control element(s), e.g., a regulatable promoter, are used such that expression can be regulated, e.g., turned on or up, or turn off or down. In some embodiments, the vector may contain a polynucleotide sequence that encodes a polypeptide, said polynucleotide sequence being in-frame with a nucleic acid inserted into said vector such that an N-terminal or C-terminal hybrid is formed. In some embodiments, the polypeptide encoded by said polynucleotide sequence may contain a signal sequence (which directs protein secretion) or a sequence which directs the expressed protein to a specific organelle or location in a cell, such as the nucleus or mitochondria. In some embodiments, the polypeptide contains a label. The label may be useful for facilitating detection and/or purification of the protein containing it. Examples of tags include a polyhistidine tag (e.g. 6X-His tag) (SEQ ID NO: 69), glutathione-8-transferase, maltose binding protein, NUS tag, SNUT tag, Strep tag, epitope tags such as V5, HA, Mus or FLAG. In some embodiments, the protease cleavage site is located in the region between the protein encoded by the inserted nucleic acid and said polypeptide, allowing said polypeptide to be removed by exposure to the protease. Vectors can be introduced into host cells using methods known in the art. One skilled in the art will be able to select an appropriate method based on, for example, vector, cell type, and the like. Examples of suitable methods include, for example, calcium phosphate mediated transfection, transfection using any of a variety of commercially available reagents, eg lipid-based or non-lipid-based, such as FuGENE, Lipofectamine, TurboFect; electroporation; bombardment with microparticles; and the like. Such methods are described in detail in standard sources, for example, Sambrook, Protocols series, and DR.

[0052] As used herein, the term "aliphatic" refers to a hydrocarbon moiety that can be straight chain (i.e., straight chain), branched, or cyclic (including fused, bridged, and spiro-fused polycyclic), and can be fully saturated, or contain one or more units of unsaturation, but is not aromatic. Unless otherwise indicated, aliphatic groups contain 1-30 carbon atoms. In some embodiments, aliphatic groups contain 1-10 carbon atoms. In other embodiments, aliphatic groups contain 1-8 carbon atoms. In other embodiments, the aliphatic groups contain 1-6 carbon atoms, and in further embodiments, the aliphatic groups contain 1^4 carbon atoms. Suitable aliphatic groups include, but are not limited to, linear or branched, alkyl, alkenyl and alkynyl groups, and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0053] As used herein, "alkyl" refers to a saturated straight, branched, or cyclic hydrocarbon containing from about 1 to about 22 carbon atoms (and all combinations and subcombinations of ranges and specific amounts of carbon atoms therein), according to some embodiments of the present invention. preferably about 1 to about 12, or about 1 to about 7 carbon atoms. Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, cyclopentyl, isopentyl, neopentyl, n-hexyl, isohexyl, cyclohexyl, cyclooctyl, adamantyl , 3-methylpentyl, 2,2-dimethylbutyl and 2,3-dimethylbutyl.

[0054] As used herein, "halogen" refers to F, Cl, Br, or I.

[0055] As used herein, "alkanoyl" refers to an optionally substituted straight or branched aliphatic acyclic moiety containing from about 1-10 carbon atoms (and all combinations and subcombinations of ranges, as well as a specific number of carbon atoms), for example, from about 1 to 7 carbon atoms, which is meant to be attached to the terminal C=O group by a single bond (and may also be referred to as an "acyl group"). Alkanoyl groups include, but are not limited to, formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, isopentanoyl, 2-methyl-butyryl, 2,2-dimethoxypropionyl, hexanoyl, heptanoyl, octanoyl, and the like; for the purposes of the present invention, a formyl group is considered an alkanoyl group. "Lower alkanoyl" refers to an optionally substituted straight or branched aliphatic acyclic moiety containing from about 1 to about 5 carbon atoms (and all combinations and subcombinations of ranges, as well as a specific number of carbon atoms). Such groups include, but are not limited to, formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, isopentanoyl, and the like.

[0056] As used herein, "aryl" refers to an optionally substituted mono- or bicyclic aromatic ring system containing from about 5 to about 14 carbon atoms (and all combinations and subcombinations of ranges and specific amounts of carbon atoms therein), preferably from about 6 up to about 10 carbon atoms. Non-limiting examples include, for example, phenyl and naphthyl.

[0057] As used herein, "aralkyl" refers to alkyl radicals bearing an aryl substituent and containing from about 6 to about 22 carbon atoms (and all combinations and subcombinations of ranges, as well as a specific number of carbon atoms), in some embodiments—preferably about 6 to about 12 carbon atoms. Aralkyl groups may optionally be substituted. Non-limiting examples include, for example, benzyl, naphthylmethyl, diphenylmethyl, triphenylmethyl, phenylethyl, and diphenylethyl.

[0058] As used herein, the terms "alkoxy" and "alkoxy" refer to an optionally substituted alkyl-O- group, where alkyl is as defined above. Examples of alkoxy groups and alkoxy groups include methoxy, ethoxy, n-propoxy, n-propoxy, n-butoxy and heptoxy.

[0059] As used herein, "carboxy" refers to the -C(=O)OH group.

[0060] As used herein, "alkoxycarbonyl" refers to -C(=O)O-alkyl

a group where alkyl is as defined above.

[0061] As used herein, "aroyl" refers to a -C(=O)-aryl group, where aryl is as defined above. Examples of aroyl groups include benzoyl and naphthoyl.

[0062] The term "cyclic ring system" refers to an aromatic or non-aromatic partially unsaturated or fully saturated 3-10-membered ring system, which includes single rings with 3-8 atoms, and bi- and tricyclic ring systems, which may include aromatic 5 - or 6-membered aryl or aromatic heterocyclic groups fused to a non-aromatic ring. Said heterocyclic rings include those containing from 1 to 3 heteroatoms independently selected from the group consisting of oxygen, sulfur and nitrogen. In certain embodiments, the term "heterocyclic" refers to a non-aromatic 5-, 6-, or 7-membered ring or polycyclic group in which at least one ring atom is a heteroatom selected from the group consisting of O, S, and N, including , but not limited to, a bi- or tricyclic group containing fused six-membered rings with one to three heteroatoms independently selected from the group consisting of oxygen, sulfur and nitrogen. In some embodiments, "cyclic ring system" refers to a cycloalkyl group, which according to the present application refers to groups with 3-10, for example, 4-7 carbon atoms. Cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like, which are optionally substituted. In some embodiments, "cyclic ring system" refers to a cycloalkenyl or cycloalkynyl moiety that is optionally substituted.

[0063] Typically, substituted chemical moieties contain one or more hydrogen substituents. Examples of substituents include, for example, halogen, alkyl, cycloalkyl, aralkyl, aryl, sulfhydryl, hydroxyl (-OH), alkoxy, cyano (-CN), carboxyl (-COOH), -C(=O)O-alkyl, aminocarbonyl ( -C(=O)NH ₂ ), -N-substituted aminocarbonyl (-C(=O)NHR''), CF ₃ , CF ₂ CF ₃ and the like. With respect to the above substituents, each R'' moiety may independently be, for example, any of H, alkyl, cycloalkyl, aryl, or aralkyl.

[0064] As used herein, "L-amino acid" refers to any of the naturally occurring left-handed alpha-amino acids commonly found in proteins or alkyl esters of said alpha-amino acids. The term "D-amino acid" refers to dextrorotatory alpha-amino acids. Unless otherwise indicated, all amino acids mentioned herein are L-amino acids.

[0065] As used herein, an "aromatic amino acid" is an amino acid containing at least one aromatic ring, such as an aryl group.

[0066] As used herein, an "aromatic amino acid analog" is an amino acid analog containing at least one aromatic ring, such as an aryl group.

[0067] II. Complement system

[0068] For ease of understanding of the present invention, and without any limitation of the present invention, this section provides a brief description of complement and its activation pathways. More information can be found, for example, in ^Kuby Immunology, 6th ed., 2006; Paul, WE, Fundamental Immunology, Lippincott Williams &Wilkins; 6th ^ed ., 2008; and Walport MJ., Complement. First of two parts. NEngl J Med., 344(14): 1058-66, 2001.

[0069] Complement is a part of the innate immune system that plays an important role in protecting the body from infectious agents. The complement system includes more than 30 serum and cellular proteins involved in three major pathways known as the classical, alternative, and lectin pathways. The classical pathway is usually initiated by binding of an antigen-IgM or IgG antibody complex to C1 (although some other activators may also initiate this pathway). Activated C1 cleaves C4 and C2 to give C4a and C4b, along with C2a and C2b. C4b and C2a combine to form C3 convertase, which cleaves C3 to form C3a and C3b. When C3b binds to C3 convertase, C5 convertase is formed, which cleaves C5 into C5a and C5b. C3a, C4a and C5a are anaphylatoxins and mediate various reactions in the acute inflammatory response. C3a and C5a are also chemotactic factors that attract cells of the immune system such as neutrophils.

[0070] The alternative pathway is initiated and amplified, for example, on the surface of microorganisms and various complex polysaccharides. In this pathway, hydrolysis of C3 to C3(H ₂ O), which occurs spontaneously at a low level, leads to the binding of factor B, which is cleaved by factor D to form a liquid-phase C3 convertase, which activates complement by cleaving C3 into C3a and C3b. C3b binds to targets such as the cell surface and forms a complex with factor B, which is later cleaved by factor D to form C3 convertase. Surface-bound C3 convertases cleave and activate additional C3 molecules, which leads to rapid deposition of C3b immediately adjacent to the site of activation, as well as additional production of C3 convertase, which in turn synthesizes additional C3b. This process leads to the launch of a cycle of C3 cleavage and the formation of C3-convertase, which significantly enhances the response. Cleavage of C3 and binding of the next C3b molecule to C3 convertase leads to the formation of C5 convertase. The C3 and C5 convertases of this pathway are controlled by the host cell molecules CR ¹ , DAF, MCP, CD59 and fH. The mechanism of action of these proteins includes either a degradation-accelerating activity (ie, the ability to degrade convertases), or the ability to function as cofactors in the degradation of C3b or C4b by factor I, or both. Normally, the presence of complement regulatory proteins on the surfaces of host cells prevents significant complement activation on them.

[0071] C5 convertases synthesized in both pathways cleave C5 to form C5a and C5b. C5b then binds to C6, C7 and C8 to form C5b-8, which catalyzes the polymerization of C9 to form the membrane attack complex (MAC) C5b-9. Said MAA is incorporated into target cell membranes and causes cell lysis. The presence of small amounts of MAA on the cell membrane can lead to a variety of consequences in addition to cell death.

[0072] The complement lectin pathway is initiated by the binding of mannose-binding lectin (MBL) and MBL-associated serine protease (MASP) to carbohydrates. The MB ¹ -1 gene (known as LMAN-1 in humans) encodes a type I integral membrane protein located in the intermediate region between the endoplasmic reticulum and the Golgi apparatus. The MBL-2 gene encodes a soluble mannose-binding protein found in serum. In the human lectin pathway, MASP-1 and MASP-2 are involved in C4 and C2 proteolysis leading to the formation of C3 convertase described above.

[0073] Complement activity is regulated by various mammalian proteins called complement control proteins (CCP/CCK) or complement activation regulatory proteins (RCA) (US Pat. No. 6,897,290). These proteins differ in ligand specificity and complement inhibition mechanism(s). They may accelerate the normal breakdown of convertases and/or function as factor I cofactors by enzymatically cleaving C3b and/or C4b into smaller fragments. CCBs are characterized by the presence of several (typically 4-56) homologous motifs known as short consensus repeats (SCRs/SCRs), complement control protein modules (PCMs), or SUSHI domains approximately 50-70 amino acids long, which contain a conserved motif comprising four linked by disulfide bonds of cysteine (two disulfide bonds), proline, tryptophan and many hydrophobic residues. The CCP family includes complement type 1 receptor (CR ¹ ; C3b:C4b receptor), complement type 2 receptor (CR2), membrane cofactor protein (MCP; CD46), decay accelerating factor (DAF), complement factor H (fH), and C4b- binding protein (C4bp). CD59 is a membrane-bound complement regulatory protein structurally unrelated to these CCBs. Complement regulatory proteins generally serve to limit the activation of complement that would otherwise occur in cells and tissues of said mammal, such as a human host. Thus, "own" cells are usually protected from the damaging effects that would occur if complement were activated on these cells. Deficiencies or defects in complement regulatory protein(s) have been implicated in the pathogenesis of a variety of complement-mediated disorders, such as those described herein.

[0074] III. Compstatin analogues

[0075] Compstatin is a cyclic peptide that binds to C3 and inhibits complement activation. US Pat. No. 6,319,897 describes a peptide with the sequence He-[Cys-Val-Val-Gln-Asp-Trp-Gly-His-His-Arg-Cys]-Thr (SEQ ID NO: 1), with a disulfide bond between two cysteines , enclosed in parentheses. It should be understood that the name "compstatin" was not used in US Pat. No. 6,319,897, but then became used in the scientific and patent literature (see, for example, Morikis, et al., Protein Sci., 7(3):619-27 , 1998) to refer to a peptide having the same sequence as SEQ ID NO: 2 described in US Pat. No. 6,319,897 but amidated at the C-terminus as shown in Table 1 (SEQ ID NO: 8). The term "comstatin" is used herein in accordance with such usage (ie, to denote the sequence of SEQ ID NO: 8). Compstatin analogs have been developed that have higher complement inhibitory activity compared to compstatin. See, for example, WO 2004/026328 (PCT/US 2003/029653), Morikis, D., et al., Biochem Soc Trans. 32(Pt 1):28-32, 2004, Mallik, B., et al., J. Med. Chem., 274-286, 2005; Katragadda, M., et al. J. Med. Chem., 49: 4616-4622, 2006; WO 2007062249 (PCT/US 2006/045539); WO 2007044668 (PCT/US 2006/039397), WO/2009/046198 (PCT/US 2008/078593); WO/2010/127336 (PCT/US 2010/033345) and description below.

[0076] Compstatin analogs can be acetylated or amidated, for example, at the N-terminus and/or C-terminus. For example, analogs of compstatin can be acetylated at the N-terminus and amidated at the C-terminus. As used in the art as used herein, the term "comstatin" and the activities of compstatin analogues described herein relative to compstatin activity refer to C-terminally amidated compstatin (Mallik, 2005, supra).

[0077] Concatemers or multimers of compstatin or its complement inhibitory analog are also suitable for the present invention.

[0078] As used herein, the term "comstatin analog" includes compstatin and any complement-inhibiting analog thereof. The term "comstatin analogue" encompasses compstatin and other compounds developed or identified from compstatin that have a complement inhibitory activity that is at least 50% of that of compstatin as assessed, for example, using any complement activation assay routinely used in the art. , or substantially similar or equivalent assays. Certain suitable assays are described in US Pat. WO 2007044668 (PCT/US 2006/039397), WO/2009/046198 (PCT/US 2008/078593); and/or WO/2010/127336 (PCT/US 2010/033345). Said assay may measure, for example, alternatively or classically mediated erythrocyte lysis, or it may be an ELISA assay. In some embodiments, the assay described in WO/2010/135717 (PCT/US 2010/035871) is used.

[0079] Compstatin analog activity can be expressed in terms of IC ₅₀ (concentration of a compound that inhibits complement activation by 50%), where a lower IC ₅₀ is known in the art to indicate higher activity. The activity of the preferred compstatin analogue for use according to the present invention is at least equal to that of compstatin. Note that certain modifications known to reduce or completely inhibit complement inhibitory activity may be expressly excluded from any embodiment of the present invention. Using an alternative pathway-mediated erythrocyte lysis assay, the IC ₅₀ for compstatin was found to be 12 μM (WO 2004/026328). It should be borne in mind that the exact IC ₅₀ value for a particular compstatin analogue when measured will vary depending on the experimental conditions (eg, the concentration in the serum used for analysis). Comparative values are used, such as those obtained from experiments in which the IC ₅₀ is determined for several different compounds under substantially identical conditions. In one embodiment, the IC ₅₀ of the compstatin analog does not exceed the IC ₅₀ of compstatin. According to certain embodiments of the present invention, the activity of said analog of compstatin is 2-99 times higher than that of compstatin (ie, the IC ₅₀ value of said analog is 2-99 times less than the IC ₅₀ value of compstatin). For example, said activity may be 10-50 times that of compstatin, or 50-99 times that of compstatin. According to certain embodiments of the present invention, the activity of the specified analogue of compstatin is 99-264 higher than that of compstatin. For example, said activity may be 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, or 264 times that of compstatin. According to certain implementation options, the specified activity is 250-300, 300-350, 350-400 or 400-500 times higher than the activity of compstatin. The present invention also encompasses analogs of compstatin with an activity greater than that of compstatin by 500-1000 times or more. In certain embodiments, the IC _{50 of} said compstatin analog is from about 0.2 μM to about 0.5 μM. In certain embodiments, the IC _{50 of} said compstatin analog is from about OD μM to about 0.2 μM. In certain embodiments, the IC _{50 of} said compstatin analog is from about 0.05 μM to about 0.1 μM. In certain embodiments, the IC _{50 of} said compstatin analog is from about 0.001 μM to about 0.05 μM.

[0080] The Kd of compstatin binding to C3 can be measured using isothermal titration calorimetry (Katragadda, et al., J. Biol. Chem., 279(53), 54987-54995, 2004). The C3 binding capacity of various compstatin analogs has been found to correlate with their activity, with lower Kd being known in the art to indicate higher binding capacity. A linear correlation between binding capacity and potency has been shown for some of the analogues tested (Katragadda, 2004, supra; Katragadda 2006, supra). According to certain embodiments of the present invention, said compstatin analog binds to C3 with a Kd value of 0.1 μM to 1.0 μM, 0.05 μM to 0.1 μM, 0.025 μM to 0.05 μM, 0.015 µM to 0.025 µM, 0.01 µM to 0.015 µM, or 0.001 µM to 0.01 µM.

[0081] Compstatin-derived or identified compounds include, but are not limited to, compounds containing an amino acid chain whose sequence is obtained by: (i) modifying the compstatin sequence (e.g., replacing one or more amino acids of said compstatin sequence with another amino acid or an amino acid analog, insertion of one or more amino acids or an amino acid analog into said compstatin sequence, or deletion of one or more amino acids from said compstatin sequence); (ii) selecting from a phage display peptide library in which one or more compstatin amino acids are randomized and optionally then modified according to method (i); or (iii) screening identification for compounds competing for binding to C3 or a fragment thereof with compstatin or any analogue thereof prepared by methods (i) or (ii). Many suitable analogs of compstatin contain a hydrophobic cluster, a beta-bend and a disulfide bridge.

[0082] According to certain embodiments of the present invention, the sequence of said compstatin analog comprises or consists essentially of a sequence obtained by making 1, 2, 3, or 4 substitutions in the compstatin sequence, i.e. substitution of 1, 2, 3 or 4 amino acids in the compstatin sequence with another standard amino acid or non-standard amino acid. The amino acid at position 4 is changed in certain embodiments of the present invention. The amino acid at position 9 is changed in certain embodiments of the present invention. The amino acids at positions 4 and 9 are changed in certain embodiments of the present invention. Only the amino acids at positions 4 are changed in certain embodiments of the present invention. and 9. According to certain embodiments of the present invention, the amino acid at position 4 or 9 is changed; or, in some embodiments, both amino acids 4 and 9 are changed, and further changed to 2 amino acids at positions selected from 1, 7, 10, 11, and 13. In certain embodiments of the present invention, the amino acids at positions 4, 7 are changed and 9. According to certain embodiments of the present invention, the amino acids at position 2 or 12, or both, are altered, provided that said alteration retains the cyclization ability of said compound. Such change(s) at positions 2 and/or 12 may be carried out along with change(s) at positions 1, 4, 7, 9, 10, 11 and/or 13. The sequence of any of these analogues compstatin, which is obtained by replacing one or more amino acids in the sequence of compstatin, optionally also contains up to 1, 2 or 3 additional amino acids at the C-terminus. In one embodiment, said additional amino acid is Gly. The sequence of any of these analogs of compstatin, which is obtained by replacing one or more amino acids in the compstatin sequence, optionally also contains up to 5 or up to 10 additional amino acids at the C-terminus. It should be understood that compstatin analogs may have any one or more of the characteristics or properties of the various embodiments described herein, and the characteristics or properties of any embodiment may further characterize any other embodiment described herein, unless indicated or apparent from context is different. According to certain embodiments of the present invention, the sequence of said compstatin analog contains or consists essentially of a sequence identical to that of compstatin except for positions corresponding to positions 4 and 9 in the compstatin sequence.

[0083] Compstatin and certain analogs of compstatin, the activity of which is higher than that of compstatin, contain exclusively standard amino acids ("standard amino acids" are glycine, leucine, isoleucine, valine, alanine, phenylalanine, tyrosine, tryptophan, aspartic acid, asparagine, glutamic acid, glutamine , cysteine, methionine, arginine, lysine, proline, serine, threonine and histidine). Certain high potency compstatin analogs contain one or more non-standard amino acids. Suitable non-standard amino acids include mono- and multi-halogenated (e.g., fluorinated) amino acids, D-amino acids, homo-amino acids, n-alkyl-amino acids, dehydroamino acids, aromatic amino acids from phenylalanine, tyrosine and tryptophan), ortho-, meta- or para-aminobenzoic acid, phosphoamino acids, methoxylated amino acids and a,a-disubstituted amino acids. In certain embodiments of the present invention, the compstatin analog is prepared by replacing one or more L-amino acids in the compstatin analog described elsewhere herein with the appropriate D-amino acid. Such compounds and methods for their use are aspects of the present invention. Examples of suitable non-standard amino acids include 2-naphthylalanine (2-NaI), 1-naphthylalanine (1-NaI), 2-indanylglycinecarboxylic acid (2Ig1), dihydrotryptophan (Dht), 4-benzoyl-L-phenylalanine (Bpa), 2-α -aminobutyric acid (2-Abu), 3-α-aminobutyric acid (3-Abu), 4-α-aminobutyric acid (4-Abu), cyclohexylalanine (Cha), homocyclohexylalanine (hCha), 4-fluoro-L-tryptophan (4fW), 5-fluoro-L-tryptophan (5fW), 6-fluoro-L-tryptophan (6fW), 4-hydroxy-L-tryptophan (40H-W), 5-hydroxy-L-tryptophan (50H-W ), 6-hydroxy-L-tryptophan (bOH-W), 1-methyl-L-tryptophan (lMeW), 4-methyl-L-tryptophan (4MeW), 5-methyl-L-tryptophan (5MeW), 7- aza-L-tryptophan (7aW), α-methyl-L-tryptophan (αMeW), β-methyl-L-tryptophan (βMeW), N-methyl-L-tryptophan (NMeW), ornithine (orn), citrulline, norleucine , γ-glutamic acid, and the like.

[0084] In certain embodiments of the present invention, said compstatin analog comprises one or more Trp analogs (eg, at position 4 and/or 7 relative to the compstatin sequence). Examples of Trp analogues are given above. See also Beene, et. al. Biochemistry 41: 10262-10269, 2002 (which includes mono- and multi-halogenated Trp analogues); Babitzke & Yanofsky, J. Biol. Chem. 270: 12452-12456, 1995 (which describes, inter alia, methylated and halogenated Trp, and other Trp and indole analogues); and US Pat. Nos. 6,214,790; 6,169,057; 5,776,970; 4,870,097; Amino acids containing two or more aromatic rings, including substituted, unsubstituted or alternatively substituted variants, are of interest as Trp analogs. According to certain embodiments of the present invention, said Trp analog, for example at position 4, is 5-methoxy, 5-methyl-, 1-methyl-, or 1-formyl-tryptophan. In certain embodiments of the present invention, a Trp analog (eg, at the 4 position) is used containing a 1-alkyl substituent, eg, a lower alkyl (eg, C ₁ -C ₅ ) substituent. In certain embodiments, N(α) methyltryptophan or 5-methyltryptophan is used. In some embodiments, an analog containing a 1-alkanoyl substituent, such as a lower alkanoyl (eg, C ₁ -C ₅ ), is used. Examples include 1-acetyl-L-tryptophan and L-β-tryptophan.

[0085] In certain embodiments, said Trp analogue has increased hydrophobicity compared to Trp. For example, the indole ring may be substituted with one or more alkyl (eg, methyl) groups. In certain embodiments, said Trp analog participates in hydrophobic interactions with C3. Such a Trp analog may be located, for example, at position 4 relative to the compstatin sequence. In certain embodiments, said Trp analog contains a substituted or unsubstituted bicyclic aromatic ring component or two or more substituted or unsubstituted monocyclic aromatic ring components.

[0086] In certain embodiments, said Trp analog has an increased tendency to form hydrogen bonds with C3 compared to Trp, but does not have an increased hydrophobicity compared to Trp. Said Trp analogue may have an increased polarity compared to Trp and/or an increased ability to participate in electrostatic interaction with a C3 hydrogen bond donor. Certain examples of Trp analogs that have an increased ability to form hydrogen bonds contain an electronegative substituent on the indole ring. Such a Trp analog may be located, for example, at position 7 relative to the compstatin sequence.

[0087] In certain embodiments of the present invention, said compstatin analog comprises one or more Ala analogs (e.g., at position 9 relative to the compstatin sequence), e.g., Ala analogs that are identical to Ala except that they contain one or more CH ₂ - groups in the side chain. In certain embodiments, said Ala analog is a straight chain amino acid with a single methyl group, such as 2-Abu. In certain embodiments of the present invention, said compstatin analog comprises one or more Trp analogs (eg, at position 4 and/or 7 relative to the compstatin sequence) and an Ala analog (eg, position 9 relative to the compstatin sequence).

[0088] According to certain embodiments of the present invention, said compstatin analog is a compound that contains a peptide having the sequence (X'aa) _n - Gln-Asp-Xaa-Gly-(X''aa) _m , (SEQ ID NO: 2), where each X'aa and each X''aa is an independently selected amino acid or amino acid analog, and Xaa is Trp or Trp analog, and n>1 and m>1 and the value of n+m is from 5 up to 21. This peptide has the core sequence Gln-Asp-Xaa-Gly (SEQ ID NO:70), where Xaa is Trp or a Trp analog, for example, a Trp analog having an increased propensity to form hydrogen bonds with an H-bond donor at compared to Trp, however, in some embodiments, does not have increased hydrophobicity compared to Trp. For example, said analogue may be one in which the Trp indole ring is substituted with an electronegative moiety, eg a halogen such as fluorine. In one embodiment, Xaa is 5-fluorotryptophan. One skilled in the art will appreciate that, in the absence of evidence to the contrary, any non-naturally occurring peptide whose sequence contains the indicated core sequence and which inhibits complement activation and/or binds to C3 will be derived from a compstatin sequence. In an alternative implementation, Xaa is an amino acid or non-Trp analog of an amino acid that allows the Gln Asp Xaa Gly peptide to form a beta-fold.

[0089] According to certain embodiments of the present invention, said peptide has the core sequence X'aa-Gln Asp Xaa Gly (SEQ ID NO: 3), where X'aa and Xaa are selected from Trp and Trp analogs. According to certain embodiments of the present invention, said peptide has the core sequence X'aa-Gln-Asp-Xaa-Gly (SEQ ID NO: 3), where X'aa and Xaa are selected from Trp, Trp analogs, and other amino acids or amino acid analogs containing at least one aromatic ring. In certain embodiments of the present invention, said core sequence forms a beta-fold in the context of a peptide. Said p-fold may be flexible, allowing the peptide to exist in two or more conformations as assessed, for example, using nuclear magnetic resonance (NMR). In certain embodiments, X'aa is a Trp analog containing a substituted or unsubstituted bicyclic aromatic ring component or two or more substituted or unsubstituted monocyclic aromatic ring components. In certain embodiments of the present invention, X'aa is selected from the group consisting of 2-naphthylalanine, 1-naphthylalanine, 2-indanylglycine carboxylic acid, dihydrotryptophan, and benzoylphenylalanine. In certain embodiments of the present invention, X'aa is a Trp analog that is more hydrophobic than Trp. For example, X'aa may be 1-methyltryptophan. In certain embodiments of the present invention, Xaa is a Trp analogue having an increased propensity to form hydrogen bonds over Trp, but in some embodiments, not having an increased hydrophobicity over Trp. According to certain embodiments of the present invention, said Trp analogue, having an increased propensity to form hydrogen bonds compared to Trp, contains a modification on the indole ring of Trp, for example, in position 5, such as the replacement of a halogen atom with an H atom in position 5. For example, Xaa may be 5-fluorotryptophan.

[0090] According to certain embodiments of the present invention, the peptide has the core sequence X'aa-Gln-Asp-Xaa-Gly-X''aa (SEQ ID NO: 4), where X'aa and Xaa are each independently selected from Trp and Trp analogs, and X''aa is selected from His, Ala, Ala analogs, Phe and Trp. In certain embodiments of the present invention, X'aa is a Trp analog that is more hydrophobic than Trp, such as 1-methyltryptophan or another Trp analog with an alkyl substituent on the indole ring (e.g., at the 1, 4, 5, or 6 position) . In certain embodiments, X'aa is a Trp analog containing a substituted or unsubstituted bicyclic aromatic ring component or two or more substituted or unsubstituted monocyclic aromatic ring components. In certain embodiments of the present invention, X'aa is selected from the group consisting of 2-naphthylalanine, 1-naphthylalanine, 2-indanylglycine carboxylic acid, dihydrotryptophan, and benzoylphenylalanine. In certain embodiments of the present invention, Xaa is a Trp analogue having an increased propensity to form hydrogen bonds with C3 relative to Trp, but in some embodiments, not having increased hydrophobicity relative to Trp. According to certain embodiments of the present invention, said Trp analogue, having an increased propensity to form hydrogen bonds compared to Trp, contains a modification on the indole ring of Trp, for example, in position 5, such as the replacement of a halogen atom with an H atom in position 5. For example, Xaa may be 5-fluorotryptophan. In certain embodiments, X''aa is Ala or an Ala analog such as Abu, or another straight chain amino acid with a single methyl group. According to certain embodiments of the present invention, said peptide has the core sequence X'aa-Gln-Asp-Xaa-Gly-X''aa (SEQ ID NO: 4), wherein X'aa and Xaa are each independently selected from Trp, Trp analogs and amino acids or amino acid analogs containing at least one aromatic side chain, and X''aa is selected from His, Ala, Ala analogs, Phe and Trp. In certain embodiments, X''aa is selected from Trp analogs, aromatic amino acids, and aromatic amino acid analogs.

[0091] According to certain preferred embodiments of the present invention, said peptide is cyclic. Said peptide may be a ring isovan bond between any two amino acids, one of which is (X'aa)n and the other is located in (X''aa)m. According to certain embodiments, the length of the cyclic part of the specified peptide is from 9 to 15 amino acids, for example, 10-12 amino acids. 12. For example, a peptide may be 13 amino acids long, with a linkage between the amino acids at positions 2 and 12 that results in an 11 amino acid cyclic moiety.

[0092] In certain embodiments, said peptide comprises or consists of the sequence X'aa1-X'aa2-X'aa3-X'aa4 -Gln-Asp-Xaa-Gly-X''aa1-X''aa2-X 'aa3-X''aa4-X''aa5 (SEQ ID NO: 5). In certain embodiments, X'aa4 and Xaa are selected from Trp and Trp analogues, and X'aa1, X'aa2, X'aa3, X''aa1, X''aa2, X''aa3, X''aa4 and X ''aa5 are independently selected from amino acids and amino acid analogs. In certain embodiments, X'aa4 and Xaa are selected from aromatic amino acids and aromatic amino acid analogs. Any one or more of X'aal, X'aa2, X'aa3, X 'aal, X''aa2, X''aa3, X''aa4 and X''aa5 may be identical to the amino acid at the corresponding position of compstatin. In one embodiment, X''aa1 is Ala or a straight chain amino acid with a single methyl group. Said peptide may be cyclized by a covalent bond between (i) X'aa1, X'aa2 or X'aa3; and (ii) X''aa2, X''aa3, X''aa4 or X''aa5. In one embodiment, said peptide is cyclized by a covalent bond between X'aa2 and X''aa4. In one embodiment, each of said covalently linked amino acids is Cys and said covalent bond is a disulfide (SS) bond. In other embodiments, said covalent bond is a C-C, C-O, CS, or CN bond. In certain embodiments, one of said covalently linked residues is an amino acid or amino acid analog with a side chain containing a primary or secondary amine, the other covalently linked residue is an amino acid or amino acid analog with a side chain containing a carboxylic acid group, and said covalent bond is is an amide bond. Amino acids or amino acid analogs with a side chain containing a primary or secondary amine include lysine and diaminocarboxylic acids with the general structure NH ₂ (CH ₂ ) _n CH(NH ₂ )COOH, such as 2,3-diaminopropionic acid (DAPC), 2, 4-diaminobutyric acid (DABA) and ornithine (orn), where n=1 (DAPA), 2 (DABA) and 3 (orn), respectively. Examples of amino acids with a side chain containing a carboxylic acid group include dicarboxylic amino acids such as glutamic acid and aspartic acid. Analogues such as beta-hydroxy-L-glutamic acid may also be used. In some embodiments, the peptide is cyclized with a thioether bond, for example as described in PCT/US 2011/052442 (WO/2012/040259). For example, in some embodiments, the disulfide bond in any of these peptides is replaced by a thioether bond. In some embodiments, cystathionine is formed. In some embodiments, said cystathionine is delta cystathionine or gamma cystathionine. In some embodiments, the modification comprises replacing the Cys-Cys disulfide bond between cysteines with X'aa2 and X''aa4 in SEQ ID NO: 5 (or at appropriate positions in other sequences) with the addition of CH ₂ , to form homocysteine at X'aa2 or X''aa4, and introducing a thioether bond to form cystathionine. In one embodiment, said cystathionine is gamma-cystathionine. In another embodiment, said cystathionine is delta cystathionine. Another modification in accordance with the present invention involves replacing the disulfide bond with a thioether bond without the addition of CH ₂ , thus forming lanthiionine. In some embodiments, a compstatin analog containing a thioether instead of a disulfide bond has improved stability, at least under some conditions, compared to a compstatin analog containing a disulfide bond.

[0093] In certain embodiments, said compstatin analog is a compound that contains a peptide having the sequence:

[0094] Xaa1-Cys-Val-Xaa2-Gin-Asp-Xaa2*-Gly-Xaa3-His-Arg-Cys-Xaa4 (SEQ ID NO: 6); where:

Xaa1 is He, Val, Leu, B ¹ -Ile, B ¹ -Val, B ¹ -Leu, or a dipeptide containing Gly-Ile or B ¹ -Gly-Ile, and B ¹ is the first blocking fragment; Xaa2 and Xaa2* are independently selected from Trp and Trp analogues; Xaa3 is His, Ala or Ala, Phe, Trp analog or Trp analog; Xaa4 is L-Thr, D-Thr, He, Val, Gly, a dipeptide selected from Thr-Ala and Thr-Asn, or a tripeptide containing Thr-Ala-Asn, with the carboxy-terminal OH of any of these L-Thr, D-Thr, He, Val, Gly, Ala or Asn is optionally replaced by a second blocking moiety B ² ; and

these two Cys residues are linked by a disulfide bond. In some embodiments, Xaa4 is Leu, Nle, His, or Phe, or a dipeptide selected from Xaa5-Ala and Xaa5-Asn, or a Xaa5-Ala-Asn tripeptide, wherein Xaa5 is selected from Leu, Nle, His, or Phe, and where the carboxy-terminal OH of any of said L-Thr, D-Thr, He, Val, Gly, Leu, Nle, His, Phe, Ala or Asn is optionally replaced by a second blocking moiety B ² ; and these two Cys residues are linked by a disulfide bond.

[0095] In other embodiments, Xaa1 is absent or is any amino acid or amino acid analog, and Xaa2, Xaa2*, Xaa3, and Xaa4 are as defined above. If Xaa1 is absent, a B ¹ blocking fragment can be attached to the N-terminal residue of Cys.

[0096] In another embodiment, Xaa4 is any amino acid or amino acid analog, and Xaa1, Xaa2, Xaa2*, and Xaa3 are as defined above. In yet another embodiment, Xaa4 is a dipeptide selected from the group consisting of: Thr-Ala and Thr-Asn, wherein the carboxy-terminal OH or Ala or Asn is optionally replaced by a second B ² blocking moiety.

[0097] In any embodiment of the compstatin analogue of SEQ ID NO: 6, Xaa2 may be Trp.

[0098] In any embodiment, the compstatin analog of SEQ ID NO: 6 Xaa2 may be a Trp analog containing a substituted or unsubstituted bicyclic aromatic ring moiety or two or more substituted or unsubstituted monocyclic aromatic ring moieties. For example, said Trp analog may be selected from 2-naphthylalanine (2-NaI), 1-naphthylalanine (1-NaI), 2-indanylglycine carboxylic acid (Ig1), dihydrotryptophan (Dht), and 4-benzoyl-L-phenylalanine.

[0099] In any embodiment, the compstatin analog of SEQ ID NO: 6 Xaa2 may be a Trp analog with increased hydrophobicity compared to Trp. For example, said Trp analog may be selected from 1-methyltryptophan, 4-methyltryptophan, 5-methyltryptophan and 6-methyltryptophan. In one embodiment, said Trp analog is 1-methyltryptophan. In one embodiment, Xaa2 is 1-methyltryptophan, Xaa2* is Trp, Xaa3 is Ala, and the other amino acids are identical to the corresponding compstatin amino acids.

[00100] In any embodiment, the compstatin analog of SEQ ID NO: 6 Xaa2* may be a Trp analog, such as a Trp analog with an increased tendency to hydrogen bond to C3 compared to Trp, which, in some embodiments, is not characterized by increased hydrophobicity compared to Trp. In certain embodiments, said Trp analog contains an electronegative substituent on the indole ring. For example, said Trp analog may be selected from 5-fluorotryptophan and 6-fluorotryptophan.

[00101] In certain embodiments of the present invention, Xaa2 is Trp, and Xaa2* is a Trp analogue with an increased tendency to form a hydrogen bond with C3 compared to Trp, which, in some embodiments, does not have an increased hydrophobicity compared to Trp . In certain embodiments, the compstatin analog of SEQ ID NO: 6 Xaa2 is a Trp analog with increased hydrophobicity over Trp, such as a Trp analog selected from 1-methyltryptophan, 4-methyltryptophan, 5-methyltryptophan, and 6-methyltryptophan, and Xaa2* is a Trp analog with an increased propensity to form a hydrogen bond with C3 compared to Trp, which, in some embodiments, does not have an increased hydrophobicity compared to Trp. For example, in one embodiment, Xaa2 is methyltryptophan and Xaa2* is 5-fluorotryptophan.

[00102] In some of the embodiments mentioned above, Xaa3 is Ala. In some of the embodiments mentioned above, Xa3 is a straight chain amino acid with a single methyl group, such as Abu.

[00103] The present invention also provides compstatin analogs with the sequence of SEQ ID NO: 6 as described above, wherein Xaa2 and Xaa2* are independently selected from Trp, Trp analogs, and other amino acids or amino acid analogs that contain at least one aromatic ring, and Xaa3 is His, Ala or Ala, Phe, Trp analog, Trp analog, or other aromatic amino acid or aromatic amino acid analog.

[00104] According to certain embodiments of the present invention, the blocking moiety present at the N- or C-terminus of any of the compstatin analogs described herein is any moiety that stabilizes the peptide, preventing its degradation that would otherwise occur in the blood or interstitial fluid a mammal (eg, a human or a non-human primate). For example, blocking fragment B ¹ can be any fragment that changes the structure of the N-terminus of the peptide in such a way as to inhibit the cleavage of the peptide bond between the N-terminal amino acid of the specified peptide and the adjacent amino acid. Blocking fragment B ² can be any fragment that changes the structure of the C-terminus of the peptide in such a way as to inhibit the cleavage of the peptide bond between the C-terminal amino acid of the specified peptide and the adjacent amino acid. Any suitable blocking fragments known in the art may be used. According to certain embodiments of the present invention, the blocking fragment B ¹ contains an acyl group (ie, the part of the carboxylic acid remaining after removal of the -OH group). Said acyl group typically contains from 1 to 12 carbon atoms, for example from 1 to 6 carbon atoms. For example, in some embodiments of the present invention, the blocking moiety B ¹ is selected from the group consisting of: formyl, acetyl, proprionyl, butyryl, isobutyryl, valeryl, isovaleryl, and the like. In one embodiment, the blocking moiety B ¹ is an acetyl group, i. e. Xaa1 is Ac-Ile, Ac-Val, Ac-Leu or Ac-Gly-Ile.

[00105] According to certain embodiments of the present invention, the blocking moiety B ² is a primary or secondary amine (-NH ₂ or NHR ¹ where R is an organic moiety such as an alkyl group).

[00106] According to certain embodiments of the present invention, the B ¹ blocking moiety is any moiety that neutralizes or reduces the positive charge that may otherwise be present at the N-terminus at physiological pH levels. In certain embodiments of the present invention, a B ² blocking moiety is any moiety that neutralizes or reduces negative charge that may otherwise be present at the C-terminus at physiological pH levels.

[00107] According to certain embodiments of the present invention, the compstatin analog is acetylated or amidated at the N-terminus and/or C-terminus, respectively. The compstatin analog can be N-terminally acetylated, C-terminally amidated and/or both N-terminally acetylated and C-terminally amidated. According to certain embodiments of the present invention, the compstatin analog contains an alkyl or aryl group at the N-terminus, and not an acetyl group.

[00108] In certain embodiments, said compstatin analog is a compound that contains a peptide having the sequence:

[00109] Xaa1-Cys-Val-Xaa2-Gin-Asp-Xaa2*-Gly-Xaa3-His-Arg-Cys-Xaa4 (SEQ ID NO: 7); where:

Xaa1 is He, Val, Leu, Ac-Ile, Ac-Val, Ac-Leu, or a dipeptide containing Gly-He or Ac-Gly-He;

Xaa2 and Xaa2* are independently selected from Trp and Trp analogues; Xaa3 is His, Ala or Ala, Phe, Trp analog or Trp analog; Xaa4 is L-Thr, D-Thr, He, Val, Gly, a dipeptide selected from Thr-Ala and Thr-Asn, or a tripeptide containing Thr-Ala-Asn, with the carboxy-terminal -OH of any of L-Thr, D-Thr, He, Val, Gly, Ala or Asn is optionally replaced by -NH ₂ ; and these two Cys residues are linked by a disulfide bond. In some embodiments, Xaa4 is Leu, Nle, His, or Phe, or a dipeptide selected from Xaa5-Ala and Xaa5-Asn, or a Xaa5-Ala-Asn tripeptide, wherein Xaa5 is selected from Leu, Nle, His, or Phe, and where the carboxy-terminal OH of any of said L-Thr, D-Thr, He, Val, Gly, Leu, Nle, His, Phe, Ala, or Asn is optionally replaced by a second blocking moiety B ² ; and these two Cys residues are linked by a disulfide bond.

[00110] In some embodiments, Xaa1, Xaa2, Xaa2*, Xaa3, and Xaa4 are as described above for various implementations of SEQ ID NO: 6. For example, in some embodiments, Xaa2* is Trp. In certain embodiments, Xaa2 is a Trp analogue with increased hydrophobicity compared to Trp, for example, 1-methyltryptophan. In certain embodiments, Xaa3 is Ala. In certain embodiments, Xaa3 is a straight chain amino acid with a single methyl group.

[00111] In certain embodiments of the present invention, Xaa1 is He and Xaa4 is L-Thr.

[00112] According to certain embodiments of the present invention, Xaa1 is He, Xaa2* is Trp, and Xaa4 is L-Thr.

[00113] g The present invention also provides compstatin analogs with the sequence of SEQ ID NO: 7 as described above, wherein Xaa2 and Xaa2* are independently selected from Trp, Trp analogs, other amino acids, or aromatic amino acid analogs, and Xaa3 is His, Ala, or an Ala, Phe, Trp analog, Trp analog, or other aromatic amino acid or aromatic amino acid analog.

[00114] In certain embodiments of any of these compstatin analogs described herein, a Phe analog is used, but not Phe.

[00115] Table 1 provides a non-limiting list of compstatin analogues suitable for the present invention. In the left column are the abbreviations of the indicated analogs, with the specific modifications noted at the marked positions (1-13) relative to the parent peptide, compstatin. As used in the art, the term "comstatin" as used herein, as well as the comparative activities of the compstatin analogs described herein versus compstatin, refers to the C-terminally amidated compstatin peptide. Unless otherwise noted, the peptides in Table 1 are amidated at the C-terminus. Bold type is used to indicate certain modifications. Activity relative to compstatin activity is based on data from published sources and the assays described therein (WO 2004/026328, WO 2007044668, Mallik, 2005; Katragadda, 2006). In cases of access to several publications, where activity is indicated, the value published later is used; it should be understood that the values may be corrected in case of discrepancies between the assay data. It should also be borne in mind that, according to some embodiments of the present invention, the peptides listed in Table 1, when used in therapeutic compositions and methods according to the present invention, are cyclized by a disulfide bond between two Cys residues. Variants of methods of cyclization of these peptides are also included in the scope of the present invention. As already noted, according to various embodiments of the present invention, one or more amino acids of a compstatin analog (eg, any of the compstatin analogs described herein) may be an N-alkyl amino acid (eg, an N-methyl amino acid). For example, without limitation, at least one amino acid in the cyclic portion of the specified peptide, at least one amino acid in the direction of the N-terminus relative to the cyclic portion and/or at least one amino acid in the direction of the C-terminus relative to the cyclic portion may be N -alkyl-amino acid, for example, N-methyl-amino acid. According to some embodiments of the present invention, the compstatin analogue contains, for example, N-methylglycine, for example, at position 8 of compstatin and/or position corresponding to position 13 of compstatin. In some embodiments, one or more of the compstatin analogs in Table 1 contains at least one N-methylglycine, for example, at position 8 of compstatin and/or position corresponding to position 13 of compstatin. In some embodiments, one or more of the compstatin analogs in Table 1 contains at least one N-methyl-isoleucine, for example, in a position corresponding to position 13 of compstatin. For example, Thr at or near the C-terminus of a peptide whose sequence is shown in Table 1, or any other compstatin analog sequence, can be replaced with N-methyl-Ile. Obviously, in some embodiments, N-methylated amino acids contain M-methyl-Gly at position 8 and N-methyl-Ilee at position 13. In some embodiments, N-methylated amino acids contain N-methyl-Gly in the core sequence, such as SEQ ID NO: 3 or SEQ ID NO: 4. In some embodiments, the N-methylated amino acids contain N-methyl-C1y in a core sequence such as SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7 .

[00117] In certain embodiments of the compositions and methods of the present invention, said compstatin analog has a sequence selected from sequences 9-36. In certain embodiments of the compositions and methods of the present invention, said compstatin analog has a sequence selected from SEQ ID NOs: 14, 21, 28, 29, 32, 33, 34, and 36. In certain embodiments of the compositions and/or of the methods of the present invention, said compstatin analog has a sequence selected from SEQ ID NOs: 30 and 31. In one embodiment of the compositions and methods of the present invention, said compstatin analog has the sequence of SEQ ID NO: 28. In one embodiment of the compositions and methods of the present invention, said compstatin analog has the sequence of SEQ ID NO: 32. In one embodiment of the compositions and methods of the present invention, said compstatin analog has the sequence of SEQ ID NO: 34. In one embodiment of the compositions and methods of the present invention, said compstatin analog has the sequence of SEQ ID NO: 34. In one embodiment of the compositions and methods, proposed in the present invention, the specified compstatin analogue has the sequence of SEQ ID NO: 36.

[00118] In some embodiments, the B ¹ blocking fragment contains an amino acid that can be represented as Xaa0. In some embodiments, the B ² blocking fragment contains an amino acid that can be represented as XaaN. In some embodiments, the B ¹ and/or B ² blocking moiety contains a non-standard amino acid such as a D-amino acid, N-alkyl amino acid (eg, N-methyl amino acid). In some embodiments, the B ¹ and/or B ² blocking fragment contains a non-standard amino acid that is an analog of the standard amino acid. In some embodiments, the amino acid analogue contains a lower alkyl, lower alkoxy, or halo substituent relative to the standard amino acid of which it is analogous. In some embodiments, the substituent is on the side chain. In some embodiments, the substituent is on an alpha carbon atom. In some embodiments, a B ¹ blocking fragment containing an amino acid, eg, a non-standard amino acid, also contains a B ^1a fragment. For example, blocking fragment B ¹ can be represented as B ^1a -Xaa0. In some embodiments, B ^1a neutralizes or reduces the positive charge that may otherwise be present at the N-terminus at physiological pH. In some embodiments, B ^1a contains or consists of, for example, an acyl group, which, for example, contains from 1 to 12 carbon atoms, for example, from 1 to 6 carbon atoms. In certain embodiments, the blocking moiety B ^1a is selected from the group consisting of: formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, and the like. In some embodiments, a blocking B ² fragment containing an amino acid, eg, a non-standard amino acid, may also contain a B ² a fragment. For example, a B ² blocking fragment can be represented as XaaN-B ^2a where N is the corresponding amino acid number (which depends on the numbering system used for the rest of the peptide). In some embodiments, B ² a neutralizes or reduces the negative charge that may otherwise be present at the C-terminus at physiological pH. In some embodiments, B ² a contains or consists of a primary or secondary amine (eg, NH ₂ ). It should be understood that the blocking activity of the B ^1a -Xaa0 and/or XaaN-B ^2a fragment can be provided by any or both of the components of the specified fragment according to various implementation options. In some embodiments, a blocking moiety or portion thereof, such as an amino acid residue, may contribute to increase the affinity of said compound for C3 or C3b and/or increase the activity of said compound. In some embodiments, the contribution to the affinity or activity of the amino acid residue may be at least as significant as the contribution to the blocking activity. For example, in some embodiments, Xaa0 and/or XaaN in B ^1a -Xaa0 and/or XaaN-B ^2a may function primarily to increase the affinity or activity of said compound, while B ^1a and/or B ² a may inhibit cleavage and/ or neutralize the charge of the peptide. In some embodiments, the compstatin analog comprises an amino acid sequence corresponding to any of SEQ ID NOs: 5-36, wherein said sequence of SEQ ID NOs: 5-36 is further extended at the N- and/or C-terminus. In some embodiments, said sequence may be represented as B ^1a -Xaa0-SEQUENCE-XaaN-B ^2a , where ''SEQUENCE'' is any of SEQ ID NOs: 5-36, where B ^1a and B ^2a can independently be present or absent. For example, in some embodiments, the compstatin analogue comprises B ^1a -Xaa0 -X'aa1-X'aa2-X'aa3-X'aa4 -Gln-Asp-Xaa-Gly-X'aal-X''aa2-X''aa3-X''aa4-X''aa5 -XaaN-B ^2a (SEQ ID NO: 71), where X'aal-X'aa2-X'aa3-X'aa4, Xaa, X'aal, X'' aa2, X'aa3, X''aa4 and X''aa5 are as above for SEQ ID NO: 5.

[00119] In some embodiments, the compstatin analog comprises B ^1a -Xaa0-Xaa1-Cys-Val -Xaa2 -Gin-Asp-Xaa2*-Gly-Xaa3 -His -Arg -Cys-Xaa4-XaaN-B ^2a (SEQ ID NO : 72) where Xaa1, Xaa2, Xaa2*, Xaa3 and Xaa4 are as specified above for SEQ ID NO: 6, or where Xaa1, Xaa2, Xaa2*, Xaa3 and Xaa4 are as specified for SEQ ID NO: 6 or SEQ ID NO:7.

[00120] In some embodiments, the compstatin analog comprises B ^1a -Xaa0-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaab-Xaa7-Xaa8-Xaa9 -Xaa1O-Xaa1l-Xaa12-Xaa13-XaaN-B ^2a (SEQ ID NO: 73), where Xaa1, Xaa2, Xaa3, Xaa4, Xaa5, Xaab, Xaa7, Xaa8, Xaa9, Xaa1O, Xaa1 1, Xaa12 and Xaa13 are identical to the amino acids at positions 1-13 of any of their sequences of SEQ ID NOS: 9-36.

[00121] In some embodiments, Xaa0 and/or XaaN, in any compstatin analog sequence, contains an amino acid that contains an aromatic ring with an alkyl substituent at one or more positions. In some embodiments, the alkyl substituent is a lower alkyl substituent. For example, in some embodiments, the alkyl substituent is a methyl or ethyl group. In some embodiments, the substituent is located in any position that does not eliminate the aromatic character of said compound. In some embodiments, the substituent is located in any position that does not eliminate the aromatic character of the ring to which said substituent is attached. In some embodiments, the substituent is located at the 1, 2, 3, 4, or 5 position. In some embodiments, Xaa0 contains an O-methyl analogue of tyrosine, 2-hydroxyphenylalanine, or 3-hydroxyphenylalanine. For the purposes of this specification, a lowercase "m" followed by a three-letter amino acid designation may be used to specifically indicate that an amino acid is an N-methyl-amino acid. For example, the designation "mGly" is used herein to refer to N-methylglycine (also sometimes referred to as sarcosine, or Sar). In some embodiments, Xaa0 is or contains mGly, Ttr, Phe, Arg, Trp, Thr, Tyr(Me), Cha, mPhe, mVal, mile, mAla, DTyr, DPhe, DArg, DTrp, DThr, DTyr(Me) , mPhe, mVal, mile, DAla, or DCha. For example, in some embodiments, the compstatin analog comprises a peptide having the sequence B ¹ -Ile-[Cys-Val-Trp(Me)-Gln-Asp-Trp-mGly-Ala-His-Arg-Cys]-mIle-B ² ( SEQ ID NO: 74) or B ¹ -Ile-[Cys-Val-Trp(Me)-Gln-Asp--Trp-mGly-Ala-His-Arg-Cys]-mIle-B2 (SEQ ID NO: 75) . The two Cys residues in the active compounds are connected by a disulfide bridge. In some embodiments, said peptide is acetylated at the N-terminus and/or amidated at the C-terminus. In some embodiments, B ¹ contains B ^1a -Xaa0 and/or B ² contains XaaN-B ^2a as described above. For example, in some embodiments, B ¹ contains or consists of Gly, mGly, Tyr, Phe, Arg, Trp, Thr, Tyr(Me), mPhe, mVal, mile, mAla, DTyr, DPhe, OTrp, DCha, Dala; and B ² contains NH ₂ , for example, the carboxy-OH in mile is changed to NH ₂ . In some embodiments, B ¹ contains or consists of mGly, Tyr, DTyr, or Tyr(Me), and B ² contains NH ₂ , for example, the carboxy-terminal -OH in mile is replaced by NH ₂ . In some embodiments, the He at position Xaa1 is replaced by Gly. Complement inhibition potency and/or C3b binding performance of selected compstatin analogs are described in WO/2010/127336 (PCT/US 2010/033345) and/or Qu, et al., Immunobiology (2012), doi: 10.1016/j.imbio. 2012.06.003.

[00122] In some embodiments, a blocking moiety or portion thereof, such as an amino acid residue, may contribute to increase the affinity of a compound for C3 or C3b, and/or increase the activity of said compound. In some embodiments, the contribution to the affinity or activity of the amino acid or amino acid analog may be more significant than the blocking activity.

[00123] In certain embodiments of the compositions and methods of the present invention, said compstatin analog has a sequence as shown in Table 1, but the Ac group is replaced with a B ¹ blocking fragment variant as described herein. In some embodiments, the MH ₂ group is replaced with a B ² blocking fragment variant as described herein.

[00124] In one embodiment, said compstatin analog binds to substantially the same region of the human C3 β chain as compstatin. In one embodiment, said compstatin analogue is a compound that binds to an approximately 40 kDa fragment of the C-terminal portion of the human C3 β chain to which compstatin binds (Soulika, A.M., et al., Mol. Immunol, 35:160, 1998; Soulika, A.M., et al., Mol. Immunol. 43(12):2023-9, 2006). In certain embodiments, said compstatin analog is a compound that binds to a compstatin binding site defined by the compstatin-C3 structure, eg, a crystal structure or an NMR-determined three-dimensional structure. In certain embodiments, said compstatin analog is a compound that is capable of replacing compstatin in the compstatin-C3 structure and forms substantially the same intermolecular contacts with C3 as compstatin. In certain embodiments, said compstatin analog is a compound that binds to a peptide binding site with the sequence shown in Table 1, e.g., SEQ ID NO: 14, 21, 28, 29, 32, 33, 34, or 36, 37, 71 , 72, 73, 74, or 75, or another compstatin analog sequence described herein in a peptide-C3 structure, for example, a crystal structure. In certain embodiments, said compstatin analog is a compound that binds to a peptide binding site having the sequence of SEQ ID NO: 30 or 31 in a peptide-C3 structure, eg, in a crystal structure. In certain embodiments, said compstatin analogue is a compound that is capable of displacing a peptide of SEQ ID NOs: 9-36, e.g., a compound that is capable of displacing a peptide of SEQ ID NOs: 14, 21, 28, 29, 32, 33 , 34, or 36, 37, 71, 72, 73, 74, or 75, or another compstatin analog sequence described herein, in a peptide-C3 structure, and forming substantially the same intermolecular contacts with C3 as said peptide. In certain embodiments, said compstatin analog is a compound that is capable of replacing a peptide of SEQ ID NO: 30 or 31 in a peptide-C3 structure and forming substantially the same intermolecular contacts with C3 as said peptide.

[00125] One of ordinary skill in the art will readily be able to determine whether the compstatin analog binds to the C-terminal portion of the C3 β-chain using routine experimental methods. For example, one skilled in the art can synthesize a photo-crosslinked version of said compstatin analog by incorporating a photo-crosslinking amino acid such as p-benzoyl-L-phenylalanine (Bpa) into said compound, e.g., at the C-terminus of said sequence (Soulika, A. M., et al, above). Optionally, additional amino acids may be included, for example an epitope tag such as a FLAG tag or an HA tag, to facilitate detection of said compound, eg using Western blotting. The compstatin analog is incubated with the indicated fragment and crosslinking is initiated. Colocalization of said compstatin analog and C3 fragment indicates binding. Surface plasmon resonance can also be used to determine if the compstatin analog binds to the compstatin binding site on C3 or a fragment thereof. One of skill in the art can use molecular modeling software to predict if a compound forms substantially the same intermolecular contacts with C3 as compstatin or a peptide with the sequence of any of the peptides listed in Table 1, for example, SEQ ID NO: 14 , 21, 28, 29, 32, 33, 34, or 36, or, in some embodiments, SEQ ID NO: 30 or 31, 37, 71, 72, 73, 74, or 75, or with another compstatin analog sequence as described in this document.

[00126] Compstatin analogs can be prepared using various artificial peptide synthesis methods known in the art, by condensation of amino acid residues, for example, in accordance with standard peptide synthesis methods, can be obtained by expression in vitro or in living cells of suitable nucleic acid coding sequences acids using methods known in the art. For example, peptides can be synthesized using standard solid phase techniques as described in Malik, supra, Katragadda, supra, WO 2004026328 and/or WO 2007062249. Potentially reactive moieties such as amino groups and carboxyl groups, reactive functional groups, and the like, using various protecting groups and techniques known in the art. See, for example, Protective Groups in Organic Synthesis, 3rd ed. Greene, T. W. and Wilts, P. G, Eds., John Wiley & Sons, New York: 1999. Peptides can be purified using standard methods such as reverse phase HPLC. Separation of peptide diastereomers, if necessary, can be performed using known methods such as reverse phase HPLC. If desired, the formulations may be lyophilized and then dissolved in a suitable solvent, such as water. The pH of the resulting solution can be adjusted, for example, to physiological pH values, using a base such as NaOH. Peptide formulations can optionally be characterized by mass spectroscopy, eg to confirm mass and/or disulfide bond formation. See, for example, Mallik, 2005, and Katragadda, 2006.

[00127] In certain embodiments, the compstatin analogue may be or comprises a cell-reactive compstatin analogue. Cell-reactive compstatin analogs are compounds that contain a fragment of the compstatin analog and a cell-reactive functional group that is capable of reacting with a functional group exposed on the cell surface, eg, under physiological conditions, to form a covalent bond. Thus, the cell-reactive analogue of compstatin becomes covalently attached to the cell. Without being bound by a particular theory, it is believed that a cell-attached analog of compstatin protects the cell from complement-mediated damage by, for example, binding to C3 (which may be in the form of C3(H ₂ O)) on the cell surface and/or in the vicinity of the cell. and inhibition of C3 cleavage and activation, and/or by binding to C3b and inhibiting its deposition on the cell or participation in the complement activation cascade. In some aspects of the present invention, isolated cells are contacted with a cell-reactive analog of compstatin ex vivo (outside the body). According to some aspects of the present invention, the cells are present in an isolated tissue or organ, for example, in a tissue or organ that will be transplanted into a subject. In some aspects of the present invention, cells are contacted with a cell-reactive compstatin analog in vivo by administering the cell-reactive compstatin analog to a subject. The cell-reactive analogue of compstatin becomes covalently attached to cells in vivo. In some aspects, the approach of the present invention protects cells, tissues, and/or organs from the detrimental effects of complement activation for at least two weeks without the need to restart treatment during that time.

[00128] In some embodiments, the present invention provides and/or uses compstatin analogues comprising a targeting moiety that binds non-covalently to a target molecule present on the surface of cells or tissues, or to an extracellular substance not attached to cells or tissues. Such compstatin analogs are referred to herein as "targeted compstatin analogs". Often, the target molecule is a protein or carbohydrate attached to the cell membrane and exposed to the cell surface. The targeting moiety directs the compstatin analog to a cell, tissue, or location susceptible to complement activation. In some aspects of the present invention, contact of the isolated cells with the targeted compstatin analog is performed ex vivo (outside the body). In some aspects of the present invention, the cells are present in an isolated tissue or organ, such as a tissue or organ that will be transplanted into a subject. In some aspects of the present invention, a targeted compstatin analogue is administered to a subject and the analogue becomes non-covalently attached to a cell, tissue, or extracellular substance in vivo. In some aspects, the approach of the present invention protects cells, tissues, and/or organs from the detrimental effects of complement activation for at least two weeks without the need to restart treatment during that time. In some embodiments, the targeted compstatin analogue contains both a targeting moiety and a cell-reactive moiety. The targeting moiety directs the compstatin analogue to, for example, a specific cell type by non-covalently binding to the molecule on such cells. The cell-reactive moiety then covalently binds to the cell or extracellular substance. In other embodiments, the targeted compstatin analog does not contain a cell-reactive moiety.

[00129] In some aspects, the compstatin analog may be or comprise a long-acting compstatin analog, wherein the long-acting compstatin analogs contain a moiety, such as polyethylene glycol (PEG), that increases the lifetime of the compound in the body (e.g., by reducing its clearance from the blood ). In some embodiments, the long acting compstatin analog does not contain a targeting moiety or a cell reactive moiety. In some embodiments, the long acting compstatin analog comprises a targeting moiety and/or a cell reactive moiety.

[00130] The compstatin analogue, optionally associated with a cell-reactive fragment or targeting fragment, can be modified by the addition of a molecule such as polyethylene glycol (PEG) or similar molecules to stabilize the compound, reduce its immunogenicity, increase its lifetime in the body, increase or reducing solubility and/or increasing resistance to degradation. PEGylation methods are well known in the art (V Veronese, F.M. & Harris, Adv. Drug Deliv. Rev. 54, 453-456, 2002; Davis, F.F., Adv. Drug Deliv. Rev. 54, 457-458, 2002); Hinds, K.D. & Kim, S.W. Adv. drug deliv. Rev. 54, 505-530 (2002; Roberts, M.J., Bentley, M.D. & Harris, J.M. Adv. Drug Deliv. Rev. 54, 459-476; 2002); Wang, Y.S. et al. Adv. drug deliv. Rev. 54, 547-570, 2002). A wide range of polymers, such as PEGs and modified PEGs, including derivatized PEGs, to which polypeptides can be conveniently attached, are described in the Nektar Catalog (Nektar Advanced Pegylation 2005-2006 Product Catalog, Nektar Therapeutics, San Carlos, CA), where detailed descriptions of suitable conjugation procedures are also provided. In yet another embodiment, the compstatin analog is fused to the Fc domain of an immunoglobulin or a portion thereof. In some other embodiments, the compstatin analog is conjugated to an albumin moiety or an albumin-binding peptide. Accordingly, in some embodiments, the compstatin analog is modified with one or more polypeptide or non-polypeptide components, for example, said compstatin analog is pegylated or conjugated to another moiety. In some embodiments, said component is not an immunoglobulin Fc domain or part thereof. The compstatin analog may be a multimer or part of a supramolecular complex, which may contain either a single species of molecular species or several different species (eg, several different analogs).

[00131] In some embodiments, the compstatin analog is a multivalent compound containing multiple compstatin analog moieties covalently or non-covalently linked to a polymer backbone or scaffold. These compstatin analog fragments may be identical or different. In certain embodiments of the present invention, said multivalent compound contains multiple instances, or copies, of a single fragment of a compstatin analog. According to other embodiments of the present invention, said multivalent compound comprises one or more instances of each of two or more non-identical compstatin analog moieties, eg, 3, 4, 5 or more different compstatin analog moieties. In certain embodiments of the present invention, the number of compstatin analog fragments ("n") is between 2 and 6. In other embodiments, n is between 7 and 20. In other embodiments, n is between 20 and 100. In other embodiments, n is between 100 and 1000. In other embodiments of the present invention, n is between 1000 and 10,000. In other embodiments, n is between 10,000 and 50,000. In other embodiments, n is between 50,000 and 100,000. implementation options, n is between 100,000 and 1,000,000.

[00132] Fragments of the compstatin analog can be attached directly to the polymer scaffold or can be attached by a connecting fragment that links the specified compstatin analog fragment and the polymer scaffold. Said connecting moiety can be attached to one compstatin analog moiety and to the polymeric scaffold. Alternatively, a plurality of compstatin analog fragments may be attached to the linking moiety such that said linker moiety attaches multiple compstatin analog moieties to the polymeric scaffold.

[00133] In some embodiments, said compstatin analog comprises a side chain amino acid containing a primary or secondary amine, such as a Lys residue. For example, a Lys residue, or a sequence containing a Lys residue, is added at the N-terminus and/or C-terminus of said compstatin analog. In some embodiments, the Lys residue is separated from the cyclic portion of said compstatin analog by a rigid or flexible spacer. Said spacer may, for example, include a substituted or unsubstituted, saturated or unsaturated alkyl chain, an oligo(ethylene glycol) chain, and/or other moieties, for example, as described in Section VI for linkers. The chain length may be, for example, from 2 to 20 carbon atoms. In other embodiments, said spacer is a peptide. The length of said peptide spacer may be, for example, between 1 and 20 amino acids, such as between 4 and 20 amino acids. Suitable spacers may contain or consist of, for example, multiple Gly residues, multiple Ser residues, or both. The side chain amino acid containing a primary or secondary amine and/or at least one amino acid in a spacer is optionally a D-amino acid. Any of a variety of polymer backbones or scaffolds may be used. For example, the polymer backbone or scaffold may be a polyamide, polysaccharide, polyanhydride, polyacrylamide, polymethacrylate, polypeptide, polyethylene oxide, or dendrimer. Suitable methods and polymer backbones are described, for example, in WO 98/46270 (PCT/US 98/07171) or WO 98/47002 (PCT/US 98/06963). In one embodiment, said polymer backbone or scaffold contains several reactive functional groups such as carboxylic acids, anhydride or succinimide groups. The specified polymer skeleton or scaffold is reacted with analogs of compstatin. In one embodiment, said compstatin analogue contains any of a variety of different reactive functional groups, such as carboxylic acids, anhydride or succinimide groups, which are reacted with suitable groups on the polymer backbone. Alternatively, monomeric units, which can be linked together to form a polymer backbone or scaffold, are first reacted with compstatin analogs and the resulting monomers polymerized. different compositions collect longer polymers.

[00134] IV. Mi bags of compstatin

[00135] The structure of compstatin is known in the art, and NMR structures are also known for a number of compstatin analogues with higher activity than compstatin (Malik, supra). Structural information can be used to design compstatin mimetics.

[00136] In one embodiment, a compstatin mimetic is any compound that competes with compstatin or any compstatin analog (e.g., the compstatin analog whose sequence is shown in Table 1) for binding to C3 or a fragment thereof (such as the 40 kDa β fragment chain to which compstatin binds). In some embodiments, said compstatin mimetic has an activity equal to or greater than that of compstatin. In some embodiments, said compstatin mimetic is more stable, orally available, or has better bioavailability than compstatin. Said compstatin mimetic may be a peptide, nucleic acid, or small molecule. In certain embodiments, said compstatin mimetic is a compound that binds to a compstatin binding site defined by the compstatin-C3 structure, eg, a crystal structure or a three-dimensional structure established by NMR experiments. In certain embodiments, said compstatin mimetic is a compound that is capable of displacing compstatin in the compstatin-C3 structure and forming substantially the same intermolecular contacts with C3 as compstatin. In certain embodiments, said compstatin mimetic is a compound that binds to a peptide binding site having the sequence shown in Table 1, for example, SEQ ID NO: 14, 21, 28, 29, 32, 33, 34, or 36, or according to some embodiments of SEQ ID NO: 30 or 31, or with another compstatin analog sequence, in a peptide-C3 structure. In certain embodiments, said compstatin mimetic is a compound that is capable of displacing a peptide with the sequence shown in Table 1, e.g., SEQ ID NO: 14, 21, 28, 29, 32, 33, 34, or 36, or in some embodiments SEQ ID NO: 30 or 31, or another compstatin analog sequence, in the peptide-C3 structure, and forming essentially the same intermolecular contacts with C3 as the specified peptide. In certain embodiments, said compstatin mimetic has a non-peptidic backbone, but contains side chains aligned with a sequence derived from the compstatin sequence.

[00137] One skilled in the art will appreciate that once a particular desired conformation of a short peptide has been confirmed, generally known methods can be used to obtain a peptide or peptide o mimetic corresponding to said conformation. See, for example, G.R. Marshall (1993) Tetrahedron 49: 3547-3558; Hruby and Nikiforovich (1991), Molecular Conformation and Biological Interactions, P. Balaram & S. Ramasehan, eds., Indian Acad, of Sci., Bangalore, PP. 429-455), Eguchi M, Kahn M., Mini Rev Med Chem., 2(5):447-62, 2002. Of particular importance to the present invention is that the structure of peptide analogs can be further adjusted to account for the influence of amino acid residues. various side chains, for example, the effects of functional groups or spatial restrictions, according to the data available in the relevant field of technology regarding compstatin and its analogues, including.

[00138] Those skilled in the art will appreciate that a peptide mimetic may be just as well suited as a peptide to achieve the particular backbone conformation and side chain functionality required for binding to C3 and inhibiting complement activation. Accordingly, it is intended to be within the scope of the present invention to prepare and use a C3 binding complement inhibitory compound using either naturally occurring amino acids, amino acid derivatives, amino acid analogs, or non-amino acid molecules capable of linking to form a backbone in a suitable conformation. A non-peptidic analog, or an analog containing peptide and non-peptidic components, is sometimes referred to herein as a "peptidomimetic" or "isosteric mimetic" to refer to substitutions or varieties of a peptide having generally similar conformational features of the backbone and/or other functional groups, thus being sufficiently similar with exemplary peptides for inhibiting complement activation. More generally, a compstatin mimetic is any compound in which the pharmacophores are located similarly to their position in compstatin, even if the backbone is different.

[00139] The use of peptidomimetics in the development of high affinity peptide analogs is well known in the art. Taking into account rotational restrictions similar to those inherent in amino acid residues in a peptide, it is possible to analyze analogs containing non-amino acid fragments and refine their conformational motifs using the Ramachandran diagram (Hruby & Nikiforovich 1991), along with other known methods.

[00140] One skilled in the art will readily be able to select suitable screening methods to identify additional compstatin mimetics and select those having the desired inhibitory activity. For example, compstatin or its analogue can be labeled (eg, with a radioactive or fluorescent label) and brought into contact with C3 in the presence of different concentrations of the test compound. The ability of said test compound to reduce compstatin analogue binding to C3 is evaluated. A test compound that significantly reduces the binding of said compstatin analogue to C3 is a potential compstatin mimetic. For example, a test compound that reduces the steady-state concentration of the compstatin analog-C3 complex, or that reduces the rate of formation of the compstatin analog-C3 complex by at least 25% or at least 50%, is a potential compstatin mimetic. One skilled in the art will appreciate that several screening options may be used. Screening compounds include natural products, aptamer libraries, phage display libraries, libraries of compounds synthesized using combinatorial chemistry, and the like. The present invention encompasses synthesizing a combinatorial library of compounds based on the core sequence described above and screening said library to identify compstatin mimetics. Any of these methods can also be used to identify novel compstatin analogs having higher inhibitory activity than the compstatin analogs tested to date. It should be appreciated that compstatin mimetics can be used in the cell-reactive compounds of the present invention, and the present invention provides such cell-reactive compstatin mimetics.

[00141] V. Cell-reactive compstatin analogs or long-acting compstatin analogs

[00142] As noted above, in accordance with certain embodiments, the present invention provides and/or uses a variety of cell-reactive analogues of compstatin. In some aspects, the cell-reactive compstatin analog comprises a compound of the formula ALM, where A is a fragment containing a cell-reactive functional group J, L is an optionally present connecting moiety, and M contains a compstatin analog moiety. Said compstatin analog fragment may, in various embodiments, include any compstatin analog, such as any of the compstatin analogs described above. The ALM formula encompasses embodiments in which AL is present at the N-terminus of said compstatin analog fragment, embodiments in which AL is present at the C-terminus of said compstatin analog fragment, embodiments in which AL is attached to the amino acid side chain of said compstatin analog fragment , and embodiments in which the same or different ALs are present at both ends of the M. It will be appreciated that in cases where certain compstatin analog(s) are present as a compstatin analog moiety in conjunction with the formula ALM, the functional group of said compstatin analog will react with the functional group L to form a covalent bond with A or L. ₂ ) (such an analog of compstatin, which can be represented by formulas oh R ¹ - (NH ₂ )), may have the formula R ¹ -NH-L- A, where a new covalent bond with L (eg, N-C) is formed and hydrogen is lost. Accordingly, the term "comstatin analog moiety" includes molecular structures in which at least one compstatin analog atom participates in the formation of a covalent bond with a second moiety, such as a side chain modification. A similar principle applies to compstatin analogue moieties present in the multivalent compound described above. In some embodiments, the blocking moiety at the N-terminus or C-terminus of the compstatin analog, such as the compstatin analog described in Section IV above, is replaced by AL in the structure of the cell-reactive compstatin analog. In some embodiments, or L contains a blocking fragment. In some embodiments, the cell-reactive analogue of compstatin has a molar activity of at least about 10%, 20%, or 30%, e.g., 30% to 40%, 30% to 50%, 30% to 60%, 30% to 70%, 30% to 80%, 30% to 90% or more of the activity of a corresponding compstatin analog having the same amino acid sequence (and, where applicable, one or more blocking moieties), but not containing cell-reactive fragment. In some embodiments wherein the cell-reactive compstatin analog comprises multiple compstatin analog moieties, said cell-reactive compstatin analog has a molar activity of at least about 10%, 20%, or 30%, e.g., 30% to 40% , from 30% to 50%, from 30% to 60%, from 30% to 70%, from 30% to 80%, from 30% to 90% or more of the total activity of these compstatin analogue fragments.

[00143] Cell-reactive fragment A according to various embodiments may include any of a variety of cell-reactive functional groups J. In general, a cell-reactive functional group may be selected at least in part based on factors such as (a) a particular functional group the group to be targeted; (b) the ability of said reactive functional group to react with the target functional group under physiologically acceptable ex vivo conditions (eg physiologically acceptable pH and osmolarity) and/or in vivo conditions (eg blood); (c) the specificity of the reaction between the specified reactive functional group and the target functional group under physiologically acceptable conditions ex vivo and/or in vivo; (d) the stability (eg, in vivo conditions) of the covalent bond formed by the reaction of said reactive functional group with the target functional group; (e) ease of synthesis of a cell-reactive analog of compstatin containing said reactive functional group, and the like. In some embodiments, a reactive functional group is selected that reacts with the target chemical group without releasing the substitutable group. In some embodiments, a reactive functional group is selected that results in the release of the substitutable group upon reaction with the target. Compounds containing such groups may be suitable, for example, for monitoring the progress and/or extent of the response. In some embodiments, the substituted group is physiologically acceptable to cells, tissues, or organs in a synthesis amount (e.g., in terms of concentration and/or absolute synthesis amount) and/or medically acceptable in a subject in an in vivo synthesis amount (e.g., in terms of concentration in corresponding bodily fluid, such as blood, and/or in terms of absolute synthesized amount). In some embodiments, the ex vivo-synthesized substituent group is at least partially removed, such as by washing the cells, or by washing or perfusing the tissue or organ, such as with saline.

[00144] In many embodiments, a cell-reactive functional group suitable for the present invention reacts with the side chain of an amino acid residue and/or with an N-terminal amino group or a C-terminal carboxyl group of a protein. In some embodiments, said cell-reactive functional group is reactive with sulfhydryl (-SH) groups that are present on the side chains of cysteine residues. In some embodiments, a maleimide group is used. Maleimide groups react with sulfhydryl groups of cysteine residues in proteins at physiological pH and form a stable thioether bond. In some embodiments, a haloacetyl group is used, such as an iodoacetyl or bromoacetyl group. Halogenacetyls react with sulfhydryl groups at physiological pH. The reaction of the iodoacetyl group proceeds with the nucleophilic substitution of iodine by a sulfur atom from the sulfhydryl group, resulting in the formation of a stable thioether bond. In other embodiments, an iodoacetamide group is used. In some embodiments, the cell-reactive functional group reacts with amino groups (-NH ₂ ) that are present at the N-terminus of proteins and in the side chains of lysine residues (s-amino group). In some embodiments, an activated ester is used, such as a succinimidyl ester (ie, an NHS ester). For example, n-hydroxysuccinimide (NHS) or its water-soluble analogue (sulfo-NHS) can be used in the synthesis, so that the resulting cell-reactive analogue of compstatin contains an NHS ester. In some embodiments, the cell-reactive functional group reacts with carboxyl (-COOH) groups that are present at the C-terminus of proteins and in the side chains of various amino acid residues. In some embodiments, the cell-reactive analog of compstatin is reactive with hydroxyl (-OH) groups that are present on various amino acid side chains and on carbohydrate moieties of glycosylated proteins.

[00145] In general, the linking portion L may include any one or more aliphatic and/or aromatic moieties that allow the formation of a stable compound linking the linked moieties. The term "stable" as used herein preferably refers to compounds having sufficient stability to allow the preparation and maintenance of the integrity of the specified connection for a sufficient period of time, for example, suitable for one or more of the purposes described herein. In some embodiments, L contains a saturated or unsaturated substituted or unsubstituted branched or unbranched aliphatic chain length of 1 to 30, 1 to 20, 1 to 10, 1 to 6, or 5 or less carbon atoms, where under length is mean the number of C atoms in the main (longest) chain. In some embodiments, said aliphatic chain contains one or more heteroatoms (O, N, S), which may be independently selected. In some embodiments, at least 50% of the backbone atoms L are carbon atoms. In some embodiments, L contains a (CH ₂ ) _n saturated alkyl moiety, where n is from 1 to 30.

[00146] In some embodiments, L contains one or more heteroatoms and has a total length of 1 to 1000, 1 to 800, 1 to 600, 1 to 400, 1 to 300, 1 to 200, 1 to 100, 1 to 50, 1 to 30 or 1 to 10 carbon atoms in the chain. In some embodiments, L comprises an oligo(ethylene glycol) moiety (-(O-CH ₂ -CH ₂ -) _n ), where n is 1 to 500, 1 to 400, 1 to 300, 1 to 200, 1 to 100 10 to 200 200 to 300 100 to 200 40 to 500 30 to 500 20 to 500 10 to 500 1 to 40 1 to 30 1 to 20 or 1 to 10.

[00147] In some embodiments, L contains an unsaturated moiety, such as -CH=CH- or -CH ₂ -CH=CH-; a moiety containing a non-aromatic cyclic ring system (eg, cyclohexyl moiety), an aromatic moiety (eg, aromatic cyclic ring system such as a phenyl moiety); an ether fragment (-C-O-C-); amide moiety (-C(=O)-N-); an ester fragment (-CO-0-); carbonyl moiety (-C(=O)-); imine fragment (-C=N-); a thioether fragment (-CS-C-); amino acid residue; and/or any fragment that can be formed during the reaction of two compatible reactive functional groups. According to certain embodiments, in one or more fragments of the connecting part or in the cell-reactive fragment, one or more hydrogen atoms (or other atoms) are independently replaced by one or a large number of fragments, for example, but not limited to, aliphatic; aromatic, aryl; alkyl, aralkyl, alkanoyl, aroyl, alkoxy; thio; F; Cl; Br; I; -NO ₂ ; -CN; _-CF3 ; -CH ₂ CF ₃ ; -CHS12; _-CH2OH ; _{-CH2CH2OH ;} -CH ₂ NH ₂ ; -CH ₂ SO ₂ CH ₃ ;-or -GRG1 where G is -O-, -S-, -NRG2-, -C(=O)-, -S(=O)-, -SO ₂ -, - С(=O)O-, -C(=O)NRG2-, -OC(=O)-, -NRG2C(=O)-, -OC(=O)O-, -OC(=O)NRG2- , -NRG2C(=O)O-, -NRG2C(=O)NRG2-, -C(=S)-, -C(=S)S-, -SC(=S)-, -SC(=S) S-, -C(=NRG2)-, -C(=NRG2)0-, -C(=NRG2)NRG3-, -OC(=NRG2)-, -NRG2C(=NRG3>, -NRG2SO ₂ -, - NRG2SO ₂ NRG3-, or -SO ₂ NRG2-, wherein each of RG1, RG2, and RG3 independently includes, but is not limited to, hydrogen, halogen, or an optionally substituted aliphatic, aromatic, or aryl moiety. if present as substituents, may optionally be attached by a linear fragment Preferably, combinations of substituents and variables provided by the present invention are used that provide the formation of stable compounds suitable for any one or more of the methods described herein, for example, suitable for treatment of one one or more disorders and/or for making contact with a cell, tissue or organ as described herein, and/or suitable as intermediates in the preparation of one or more of such compounds.

[00148] L according to various implementation options may include any one or more of the fragments described in the previous paragraph. In some embodiments, L contains two or more different moieties linked together to form a structure, typically 1 to about 60 atoms long, 1 to about 50 atoms, e.g., 1 to 40, 1 to 30 , 1 to 20, 1 to 10, or 1 to 6 atoms, where length refers to the number of atoms in the main (longest) chain. In some embodiments, L comprises two or more different moieties linked together to form a structure, typically containing 1 to about 40, such as 1 to 30, such as 1 to 20, 1 to 10, or 1 to 6 carbon atoms in the main (longest) chain. In general, the structure of such a cell-reactive analog of compstatin can be represented by the formula A-(L ^Pj )jM, where the value of j is typically from 1 to 10, and each is independently selected from the fragments described in the previous paragraph. In many embodiments, L contains one or more carbon-containing chains, such as -(CH ₂ )n- and/or -(O-CH ₂ -CH ₂ -)n, which(s) are covalently attached to each other and /or with a cell-reactive functional group or analogue compstatin, for example, fragments (for example, amide fragments, fragments of esters or ethers) formed by the reaction of two compatible reactive functional groups. In some embodiments, L contains an oligo(ethylene glycol) moiety and/or a saturated alkyl chain. In some embodiments, L contains -(CH ₂ ) _m -C(=O)-NH-(CH ₂ CH ₂ O) _n (CH ₂ )PC(=O)- or -(CH ₂ ) _m -C(= O)-NH-(CH ₂ ) _p (OCH ₂ CH ₂ ) _n C(=O)-. In some embodiments, m, n, and p are selected such that the number of carbon atoms in said chain is from 1 to 500, e.g., 2 to 400, 2 to 300, 2 to 200, 2 to 100, 2 up to 50, 4 to 40, 6 to 30, or 8 to 20. In some embodiments, m is 2 to 10, n is 1 to 500, and/or p is 2 to 10. In some embodiments, p is 2 to 10. in embodiments, m is 2 to 10, n is 1 to 400, and/or p is 2 to 10. In some embodiments, m is 2 to 10, n is 1 to 300, and/or the value of p is from 2 to 10. In some embodiments, the value of m is from 2 to 10, the value of n is from 1 to 200, and/or the value of p is from 2 to 10. In some embodiments, the value of m is from 2 to 10, the value of n is from 1 to 100 and/or the value of p is from 2 to 10. Consistent but in some embodiments, the value of m is from 2 to 10, the value of n is from 1 to 50, and/or the value of p is from 2 to 10. In some embodiments, the value of m is from 2 to 10, the value of n is from 1 to 25, and /or the value of p is from 2 to 10. In some embodiments, the value of m is from 2 to 10, the value of n is from 1 to 8, and/or the value of p is from 2 to 10. Optionally, at least one -CH ₂ - is replaced to CH-R, where R can be any substituent. Optionally, at least one -CH ₂ - is replaced by a heteroatom, a cyclic ring system, an amide, ester or ether moiety. In some embodiments, L does not include an alkyl group containing more than 3 carbon atoms in the longest chain. In some embodiments, L does not include an alkyl group containing more than 4, 5, 6, 7, 8, 9, 10, or 11 carbon atoms in the longest chain.

[00149] According to some embodiments of the present invention, A contains a cell-reactive functional group J and a linker L ¹ containing a connecting part L ^P1 and a reactive functional group that reacts with a compstatin analog to form AM. In some embodiments, a bifunctional L ² linker is used containing two reactive functional groups and an L ^P2 linking moiety. The reactive functional groups from L react with the appropriate reactive functional groups from A and M to form a cell-reactive analog of compstatin ALM. In some embodiments, said compstatin analog comprises an L ³ linker containing an L ^P3 connecting moiety. For example, as described below, a linker containing a reactive functional group may be present at the N- or C-terminus, or a fragment containing a reactive functional group may be attached to the N- or C-terminus by a linker. Accordingly, L may contain several connecting parts of L ^p introduced, for example, A, the linker(s) used to combine A and M, and/or a compstatin analogue. It should be understood that if present in the ALM structure, certain reactive(s) functional group(s) present(s) prior to the reaction in L ¹ , L ² , L ³ etc. will pass( ut) through the reaction, so that only part(s) of the specified(s) reactive(s) functional group(s) will(ut) be present in the final ALM structure, and the specified compound will contain fragments formed during the reaction of the specified functional groups. In general, if the connection contains two or more connecting parts, these connecting parts may be the same or different, and may be independently selected according to various implementation options. Several connecting parts L ^p can be connected to each other to form a larger connecting part L, and at least some of these connecting parts can be attached to one or more analogs of compstatin and/or one or more cell-reactive functional groups. In molecules containing multiple compstatin analogs, said compstatin analogs may be the same or different, and if they are different, may be independently selected. The same applies to connecting parts and reactive functional groups. The present invention encompasses the use of multivalent analogs of compstatin containing one or more cell-reactive functional groups, and the use of concatemers of analogs of compstatin containing one or more cell-reactive functional groups. In some embodiments, at least one bond is a stable non-covalent bond, such as a biotin/(strept)avidin bond, or other non-covalent bond of approximately equivalent strength.

[00150] In some embodiments, a cell-reactive compstatin analog comprises a compstatin analog wherein any of SEQ ID NOs: 3-36, 71, 72, 73, 74, or 75 is extended with one or more amino acids at the N-terminus , C-terminus, or both ends, wherein at least one of said amino acids has a side chain containing a reactive functional group such as a primary or secondary amine, a sulfhydryl group, a carboxyl group (which may be present as a carboxylate group), a guanidino group , a phenolic group, an indole ring, a thioether, or an imidazole ring. In some embodiments, said amino acid(s) is/are L-amino acid(s). In some embodiments, any one or more of said amino acids is a D-amino acid. If multiple amino acids are added, said amino acids can be independently selected. In some embodiments, said reactive functional group (eg, a primary or secondary amine) is used as a target for adding a cell-reactive functional group-containing moiety. Amino acids with a side chain containing a primary or secondary amine include lysine (Lys) and diaminocarboxylic acids with the general structure NH ₂ (CH ₂ ) _n CH(NH ₂ )COOH, such as 2,3-diaminopropionic acid (DAPC), 2, 4-diaminobutyric acid (DABA) and ornithine (orn), where n=1 (DAPA), 2 (DABA) and 3 (orn), respectively. In some embodiments, at least one amino acid is cysteine, aspartic acid, glutamic acid, arginine, tyrosine, tryptophan, methionine, or histidine. Cysteine contains a side chain containing a sulfhydryl group. Aspartic acid and glutamic acid contain a side chain containing a carboxyl group (capable of ionization to a carboxylate group). Arginine contains a side chain containing a guanidino group. Tyrosine contains a side chain containing a phenolic group (capable of ionization to a phenolate group). Tryptophan contains a side chain containing an indole ring. Methionine contains a side chain containing a thioether group. Histidine contains a side chain containing an imidazole ring. A wide range of non-standard amino acids are available containing side chains containing one or more of these reactive functional groups, including naturally occurring amino acids and amino acids not found in nature. See, for example, Hughes, B. (ed.), Amino Acids, Peptides and Proteins in Organic Chemistry, Volumes 1-4, Wiley-VCH (2009-2011); Blaskovich, M., Handbook on Syntheses of Amino Acids General Routes to Amino Acids, Oxford University Press, 2010. The present invention encompasses embodiments in which one or more non-standard amino acids are used to add a cell-reactive functional group-containing moiety to the backbone. Any one or more of said amino acids may be suitably protected during the synthesis of said compound. Thus, one or more amino acids can be protected during the reaction(s) involving(their) side chain of the target amino acid (target). In some embodiments that are characterized in that a sulfhydryl-containing amino acid is used as the basis for adding the cell-reactive functional group-containing moiety, said sulfhydryl is protected during cyclization of said compound by forming an intramolecular disulfide bond between other amino acids, e.g., cysteines .

[00151] In the description contained in this paragraph, an amino acid with a side chain containing an amino group is given as an example. The present invention covers similar embodiments when an amino acid with a side chain containing other reactive functional groups is used. In some embodiments, an amino acid with a side chain containing a primary or secondary amine is attached directly to the N-terminus or C-terminus of any of SEQ ID NOs: 3-36, 37, 71, 72, 73, 74, or 75, or by a peptide bond . In some embodiments, an amino acid with a side chain containing a primary or secondary amine is attached to the N- or C-terminus of any of SEQ ID NOs: 3-36, 37, 71, 72, 73, 74, or 75, or connecting (linker) a part that may contain any one or more of the connecting fragments described above. In some embodiments, at least two amino acids are added at one or both ends. Said two or more added amino acids may be linked to each other by peptide bonds, or at least some of said added amino acids may be linked to each other by a connecting portion, which may contain any one or more connecting moieties described herein. Accordingly, in some embodiments, the cell-reactive compstatin analog comprises a compstatin analog fragment M of the formula B1-R1-M ₁ -R2-B2, where M ₁ is any of the sequences of SEQ ID NOs: 3-36, 37, 71, 72, 73, 74 or 75, or R1 or R2 may be absent; at least one R1 and R2 contains an amino acid with a side chain containing a primary or secondary amine; and B ¹ and B ² are optionally present blocking fragments. R1 and/or R2 may/may be attached to Mi by a peptide bond or a non-peptide bond. R1 and/or R2 may/may contain a connecting part L ^P3 . For example, R ¹ may be of the formula M ₂ -L ^P3 u/or R2 may be of the formula L ^P3 - M ₂ where L ^P3 is a connecting moiety and M ₂ contains at least one amino acid with a side chain containing a primary or secondary amine. For example, Mg may be Lys or an amino acid chain containing Lys. In some embodiments, L ^P3 contains one or more amino acids or is composed of one or more amino acids. For example, the length of L ^P3 may be from 1 to about 20 amino acids, for example, from 4 to 20 amino acids. In some embodiments, L ^P3 contains several Gly, Ser and/or Ala residues, or consists of several Gly, Ser and/or Ala residues. In some embodiments, L ^P3 does not include amino acids containing a reactive SH group, such as Cys. In some embodiments, L ^P3 contains an oligo(ethylene glycol) moiety and/or a saturated alkyl chain. In some embodiments, L ^P3 is attached to the N-terminal amino acid Mi by an amide bond. In some embodiments, L ^P3 is attached to the C-terminal amino acid Mi by an amide bond. The specified connection can then be extended at either or both ends by adding the next(their) connecting(their) portion(s) and/or amino acid(s). These amino acids may be the same or different; if the amino acids are different, they can be chosen independently. In some embodiments, two or more amino acids with side chains containing reactive functional groups are used, characterized in that said reactive functional groups may be the same or different. These two or more reactive functional groups can be used as a basis for adding two or more fragments. In some embodiments, two or more cell-reactive fragments are added. In some embodiments, a cell-reactive fragment and a targeting fragment are added. In some embodiments, after incorporation of said amino acid into the peptide chain, a linker and/or a cell-reactive fragment is attached to the side chain of the amino acid. In some embodiments, the linker and/or cell-reactive fragment is already attached to the side chain of the amino acid prior to the use of said amino acid in the synthesis of the cell-reactive analogue of compstatin. For example, a Lys derivative having a linker attached to its side chain can be used. Said linker may contain a cell-reactive functional group or may later be modified to contain a cell-reactive functional group.

[00152] A more detailed description of certain cell-reactive analogs of compstatin is provided below. In the description below, the peptide with the amino acid sequence Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr (SEQ ID NO: 37) (corresponding to the compstatin analogue with the sequence of SEQ ID NO: 28, where the asterisks in SEQ ID NO: 37 denote cysteines in the active compound connected by a disulfide bond, and (1Me)Trp means 1-nontype-tryptophan)) are used as a typical compstatin analogue fragment; mapeimide (abbreviated as Mai) is used as an example of a cell-reactive functional group; (CH ₂ ) _n and (O-CH ₂ -CH ₂ ) _n are used as examples of connecting parts; lysine is used as an example of an amino acid containing a reactive functional group (in some compounds), and acetylation and amidation of the N- and C-terminus, respectively, are used as optionally present exemplary blocking moieties in some compounds and are shown in italics, i.e. Ac and NH ₂ respectively. It should be borne in mind that these compounds can be obtained using a variety of synthetic methods and using a variety of precursors. The discussion of various synthetic methods and precursors below is not intended to limit the present invention. In general, any property of any of the compounds described below or herein may be combined without limitation with the property(s) of other compounds described below or herein, and the present invention encompasses such embodiments.

[00153] In some embodiments, the cell-reactive moiety is provided with a cell-reactive compound containing a maleimide group (as the cell-reactive functional group) and an alkanoic acid (RCOOH) where R is an alkyl group. For example, 6-maleimido caproic acid (Mal-(CH ₂ ) ₅ -COOH) shown below can be used.

[00154] In some embodiments, the cell-reactive moiety is provided by an apanoic acid derivative that contains an activated carboxylic acid moiety, such as an OH moiety, that has been converted to a better substituent group. For example, the carboxyl group of compound I can be reacted with EDA, followed by reaction with NHS (which may optionally be a water-soluble sulfo-NHS), to give an ester derivative of 6-maleimidocaproic acid N-hydroxysuccinimide, i.e. the N-hydroxysuccinimide (NHS) ester of 6-maleimidohexanoic acid (pictured below).

[00155] The connection with the sequence of SEQ ID NO: 37 can be modified at the N- and/or C-terminus to obtain a cell-reactive analogue of compstatin. For example, compound II can be used to synthesize the following cell-reactive analogue of compstatin by reaction with the N-terminal amino group of He.

[00156] Maleimide-(CH ₂ ) ₅ -C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 38). It will be appreciated that in SEQ ID NO: 38, the -C(=O) fragment is attached directly to the C-terminal amino acid (He) by a CN bond, where N is part of the indicated amino acid and is not shown.

[00157] In other embodiments, the maleimide group is fused to Thr at the C-terminus to form the following cell-reactive analog of compstatin:

[00158] Ac-I1e-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-(C=O)-(CH ₂ ) ₅ -maleimide ( SEQ ID NO: 39).

[00159] In some embodiments, a cell-reactive analog of compstatin can be synthesized using a bifunctional linker (eg, a heterobifunctional linker). An exemplary heterobifunctional linker containing fragments (CH ₂ -CH ₂ -O) _n and (CH ₂ ) _m (where m=2) is shown below:

[00160] Compound III contains a maleimide group as a cell-reactive functional group and an NHS ester moiety that readily reacts with an amino group (eg, an N-terminal amino group or an amino side chain amino group of an amino acid).

[00161] An embodiment of compound III, where n=2, can be used to synthesize the following cell-reactive compstatin analog using the compstatin analog of SEQ ID NO: 37:

[00162] Maleimide-(CH ₂ ) ₂ -C(=O)-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln -Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 40)

[00163] It should be noted that in connection with the sequence of SEQ ID NO: 40, the -C(=O) fragment is attached to the N-terminal amino acid (He residue) by a CN bond, where N is part of the specified amino acid and is not shown. In some embodiments, the linker has the formula Compound W, where n>1. Examples of n values for the fragment (CH ₂ -CH ₂ -O)P are given in this document.

[00164] In some embodiments, the alkyl chain connecting the maleimide moiety to the rest of the molecule contains more or less methylene units, the oligo(ethylene glycol) moiety contains more or less ethylene glycol units, and/or said oligo(ethylene glycol) moiety is flanked by more or less fewer methylene units on either or both sides, compared to a compound of SEQ ID NO: 39 or SEQ ID NO: 40. Examples of cell-reactive analogs of compstatin, illustrating some of these options, are given below (SEQ ID NO: 41 -46):

[00165] Maleimide-(CH ₂ ) ₂ -C(=O)-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln -Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 41)

[00166] Maleimide-(CH ₂ ) ₃ -C(=O)-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln -Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 42)

[00167] Maleimide-(CH ₂ ) ₅ -C(=O)-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln -Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 43)

[00168] Maleimide-(CH ₂ ) ₄ -C(=O)-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln -Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 44)

[00169] Maleimide-(CH ₂ ) ₄ -C(=O)-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln -Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 45)

[00170] Maleimide-(CH ₂ ) ₅ -C(=O)-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ C(=O)-Ile-Cys*-Val-(1Me)Trp-Gln -Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 46)

[00171] In some embodiments, SEQ ID NO: 37 is extended by including a Lys residue at the N- or C-terminus of said peptide, for example, following the C-terminal bond example below:

[00172] Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-Lys-NH ₂ (SEQ ID NO: 47).

[00173] In some embodiments, a Lys residue is attached to the N- or C-terminus of SEQ ID NO: 37 with a peptide linker, such as in the C-terminal bond example below:

[00174] Ac-Ile-Cys*-Val-(1Me)-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-(Gly) ₅ -Lys-NH ₂ (SEQ ID NO: 48 ).

[00175] In some embodiments, a linker containing a primary or secondary amine is added to the N- or C-terminus of the compstatin analog. In some embodiments, said linker contains an alkyl chain and/or an oligo(ethylene glycol) moiety. For example, NH ₂ (CH ₂ CH ₂ O) _n CH ₂ C(=O)OH (for example, 8-amino-3,6-dioxaoctanoic acid (AEEAc) or 11-amino-3,6,9-trioxaundecanoic acid) can be used. acid) or its NHS ester (for example, the NHS ester of 8-amino-3,6-dioxaoctanoic acid or 11-amino-3,6,9-trioxaundecanoic acid). In some embodiments, the resulting compound is represented as follows (linker portion in bold):

[00176] NH ₂ (CH ₂ ) ₅ C(=O)-Ile-Cys-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys-Thr-NH ₂ (SEQ ID NO: 49)

[00177] NH ₂ (CH ₂ CH ₂ O) ₂ CH ₂ C(=O)-Ile-Cys-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys-Thr- NH ₂ (SEQ ID NO: 50)

[00178] In some embodiments, a Lys residue is attached to the N- or C-terminus of SEQ ID NO: 37 with a linker containing a non-peptide moiety. For example, said linker may contain an alkyl chain, an oligo(ethylene glycol) chain, and/or a cyclic ring system. In some embodiments, 8-AEEAc or its NHS ester is used to give (if Lys is attached at the C-terminus) the following compound (the 8-AEEAc part is in bold):

[00179] Ac-Ile-Cys*-Val-(1Me)-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr -NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ -C( =O)-Lys-NH ₂ (SEQ ID NO: 51)

[00180] It should be noted that in SEQ ID NOs: 49 and 50, the -C(=O) fragment is attached to an adjacent He residue by a C-N bond, where N is part of the indicated amino acid and is not shown. Similarly, in SEQ ID NO: 51, a -C(=O) fragment is attached to an adjacent Lys residue by a C-N bond, where N is part of the indicated amino acid and is not shown. It should also be noted that in SEQ ID NO: 51, the NH moiety is directly attached to the N-terminal amino acid (Thr) by a C-N bond, where C is the carbonyl carbon atom of the indicated amino acid and is not shown.

[00181] Compounds with the sequences of SEQ ID NO: 47-51 can easily be modified at the primary amino group to obtain a cell-reactive analogue of compstatin. For example, compounds of SEQ ID NOs: 47-51 (or other compounds containing a primary or secondary amine and a compstatin analog fragment) can be reacted with 6-maleimidocaproic acid N-succinimidyl ester to produce the following cell-reactive compstatin analogs:

[00182] Ac-Ile-Cys*-Val-(1Me)-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-Lys-(C(=O)-(CH ₂ ) ₅ - Mal-NH ₂ (SEQ ID NO: 52)

[00183] Ac-Ile-Cys*-Val-(1Me)-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-(Gly) ₅ -Lys--(C(=O)- (CH ₂ ) ₅ -Mal-NH ₂ (SEQ ID NO: 53)

[00184] Mal-(CH ₂ ) ₅ -(C(=O)-NH(CH ₂ ) ₅ C(=O)-Ile-Cys-Val-(1Me)-Gln-Asp-Trp-Gly-Ala- His-Arg-Cys-Thr-(Gly) ₅ -Lys--(C(=O)-(CH ₂ ) ₅ -Mal-NH ₂ (SEQ ID NO: 54)

[00185] Mal-(CH ₂ ) ₅ -(C(=O)-NH(CH ₂ CH ₂ O)CH ₂ C(=O)-Ile-Cys-Val-(1Me)-Gln-Asp-Trp- Gly-Ala-His-Arg-Cys-Thr-NH ₂ (SEQ ID NO: 55)

[00186] Ac-Ile-Cys*-Val-(1Me)-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr -NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ -C( =O)-Lys-(C(=O)-(CH ₂ ) ₅ -Mal-NH ₂ (SEQ ID NO: 56)

[00187] In another embodiment, the cell-reactive analog of compstatin is as follows: Ac-Ile-Cys*-Val-(1Me)-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-Lys-( C(=O)-CH ₂ (OCH ₂ CH2) ₂ NH(C(=O)-(CH ₂ ) ₅ -Mal-NH ₂ (SEQ ID NO: 57)

[00188] The present invention provides sequence variants of SEQ ID NOs: 38-57, where -Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr- (SEQ ID NO:76) is replaced with an amino acid sequence containing the amino acid sequence of any other compstatin analog, for example, according to any of SEQ ID NOs 3-27 or 29-36, 37, 71, 72, 73, 74, or 75, provided blocking moiety(s) present(s) at the N- and/or C-terminus of the compstatin analog may be absent, replaced by a linker (which may contain a blocking moiety) or attached to another N- or C- terminal amino acid present in the corresponding(their) option(s).

[00189] Other bifunctional cross-linkers containing maleimide as a cell-reactive moiety and an NHS ester as an amino-reactive moiety suitable for various embodiments of the present invention include, for example, succinimidyl 4-(p-maleimidophenyl)butyrate (SMPB); succinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (SMCC); ester of N-γ-maleimidobutyryl-oxysuccinimide (GMBS). Addition of a sulfonate to the NHS ring gives water-soluble analogs such as sulfosuccinimidyl(4-iodoacetyl)-aminobenzoate (sulfo-SIAB), sulfo-succinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (sulfo-SMCC), sulfosuccinimidyl 4-(p-maleimidophenyl)butyrate (sulfo-SMPB), sulfo-N-γ-maleimidobutyryloxysuccinimide ester (sulfo-GMBS) and the like, which can avoid the need to use an organic solvent. In some embodiments, a long chain version of any of the above analogues is used, containing a spacer between the NHS ester moiety and the remainder of said molecule. Said spacer may contain, for example, an alkyl chain. An example is succinimidyl-4-[N-maleimidomethyl]cyclohexane-l-carboxy-[6-amidocaproate].

[00190] In some embodiments, a bifunctional linker is used containing an NHS ester (as an amino-reactive moiety) and an iodoacetyl group (reactive with sulfhydryl groups). Such linkers include, for example, n-succinimidyl (4-iodoacetyl)-aminobenzoate (SIAB); succinimidyl 6-[(iodoacetyl)-amino]hexanoate (SIAX); succinimidyl 6-[6-(((iodoacetyl)amino)-hexanoyl)amino]hexanoate (SIAXX); succinimidyl 4-((iodoacetyl)amino)methyl)-cyclohexane-1-carboxylate (SIAC); succinimidyl 6-((((4-(iodoacetyl)amino)methyl-cyclohexane-1-carbonyl)amino)hexanoate (SIACX);

[00191] In some embodiments, a bifunctional linker is used containing an NHS ester (as an amino-reactive moiety) and a pyridyl disulfide group (as a cell-reactive moiety reactive with sulfhydryl groups). Examples include N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP); succinimidyloxycarbonyl-α-methyl-α-(2-pyridyldithio)toluene (SMPT) and variants containing a sulfonate on the NHS ring and/or a spacer containing an alkyl chain between the NHS ester moiety and the rest of the molecule (e.g., succinimidyl 6-( 3-[2-pyridyldithio]-propionamido)hexanoate) (LC-SPDP). Variants of such linkers containing additional or different moieties may be used. For example, a longer or shorter alkyl chain may be used for the spacer, or an oligo(ethylene glycol) moiety may be used in place of the alkyl chain.

[00192] In general, a cell-reactive analogue of compstatin can be synthesized using a variety of methods. Cell reactive compounds containing a cell reactive functional group and a linker are often commercially available as ready-to-use formulations. For example, 6-maleimidocaproic acid and 6-maleimidocaproic acid N-hydroxysuccinimide ester are available from various vendors. Alternatively, such compounds can be synthesized using methods known in the art. See, for example, Keller O., Rudinger J. Helv Chim Acta. 58(2):531-41, 1975 and Hashida S, et al., J Appl Biochem., 6(l-2):56-63, 1984. See also Hermanson, G. supra, and references cited. , for descriptions of methods and reagents suitable for the synthesis of conjugates. In general, the present invention encompasses any method for preparing a compound containing a compstatin analog moiety and a cell-reactive functional group, and the resulting compounds.

[00193] In some embodiments, linear peptide synthesis uses an amino acid containing a linker attached to a side chain. Said linear peptide can be synthesized using standard peptide synthesis methods known in the art, for example, using standard solid phase peptide synthesis. The linear peptide is then cyclized (eg, by oxidation of Cys residues to form an intramolecular disulfide). Then, the reaction of the specified cyclic compound with a linker containing a cell-reactive functional group can be carried out. In other embodiments, the fragment containing the cell-reactive functional group is reacted with a linear compound prior to its cyclization. In general, the reactive functional groups may be suitably protected to avoid unnecessary reaction between them during the synthesis of the cell-reactive analog of compstatin. The cell-reactive functional group, any of the amino acid side chains and/or either or both ends of said peptide may be protected during said reaction and then the protection may be deprotected. For example, the SH groups of Cys residues and/or SH-reactive moieties such as maleimides can be protected until the end of the cyclization to avoid reaction between them. The reaction conditions are selected at least in part taking into account the conditions necessary for a particular reactive(s) functional group(s) to obtain a satisfactory yield in an acceptable period of time. The temperature, pH and concentration of the reactants can be adjusted to provide the desired degree or rate of reaction. See, for example, Hermanson, supra. The desired product may be purified, for example, by removing unreacted cell-reactive functional group-containing compound, unreacted compstatin analog, linker(s), products other than the desired compstatin cell-reactive analog that may have formed during the reaction, other substances, present in the reaction mixture, and the like. Compositions and methods for preparing cell-reactive analogs of compstatin, and intermediates in their synthesis, are aspects of the present invention.

[00194] According to some aspects of the present invention, the above-described linker(s) are used in the preparation of analogs of compstatin containing a fragment such as a polyethylene glycol (PEG) chain or other polymer(s), for example, stabilizing the specified connection, increasing the time its life in the body, increasing its solubility, reducing immunogenicity and/or increasing its resistance to degradation. Without limiting the present invention in any way, such a fragment may be referred to herein as a "clearance-reducing fragment" (GFR), and a compstatin analog containing such a fragment may be referred to as a "long-acting compstatin analog" (LACA). In some embodiments, the mean plasma half-life of the long-acting compstatin analogue is at least 1 day, e.g., 1-3 days, 3-7 days, 7-14 days, or 14-28 days, when administered intravenously at a dose of 10 μg/kg in humans or non-human primates, or at a dose of about 1-3 μg/kg, 3-5 μg/kg, 5-10 μg/kg, eg 7 μg/kg. In some embodiments, the mean plasma half-life of the long-acting compstatin analog is at least 1 day, such as 1-3 days, 3-7 days, 7-14 days, or 14-28 days, when administered subcutaneously, such as at a dose of about 1-3 µg/kg, 3-5 µg/kg, 5-10 µg/kg, eg 7 µg/kg, to humans or non-human primates. In some embodiments, the mean plasma half-life (e.g., terminal half-life) of the long-acting compstatin analog is from about 4-10, 5-9, 5-8, 6-9, 7-9, or 8-9 days, e.g., approximately 4; 4.5; 5; 5.5; 6; 6.5; 7; 7.5; eight; 8.5; 9; 9.5 or 10 days when administered intravenously, eg at a dose of about 1-3 µg/kg, 3-5 µg/kg, or 5-10 µg/kg, eg 7 µg/kg, to humans or non-humans primates. In some embodiments, the mean plasma half-life (e.g., terminal half-life) of the long-acting compstatin analogue is between about 4-10, 5-9, 5-8, 6-9, 7-9, or 8-9 days, e.g., about 4;4 ,5; 5;5.5; 6; 6.5; 7; 7.5; eight; 8.5; 9; 9.5 or 10 days, when administered subcutaneously, for example, at a dose of approximately 1-3 μg/kg, 3-5 μg/kg, 5-10 μg/kg, for example, 7 μg/kg, to humans or non-humans primate people. In certain embodiments, a long acting compstatin analog is characterized in that it is extensively absorbed from the site of administration over a period of time following subcutaneous injection and provides, for example, at or about 1-2 days after administration, a blood level comparable to that can be achieved by administering the same amount of compound intravenously. In some embodiments, the indicated blood level at or about 2, 3, 4, 5, 6, 7, 8 or more days after the subcutaneous dose is within about 5%, 10%, 15%, 20%, or 25%. on the blood level that can be achieved by administering the same amount of the compound intravenously. See, for example, FIG. 11 showing the pharmacokinetics of an intravenous and subcutaneous dose for an exemplary compound described herein approximately 1-2 days after administration. In some embodiments, the mean plasma half-life of a long-acting compstatin analog when administered intravenously at a dose of 10 μg/kg to humans or non-human primates is increased by at least 2-fold, e.g., 25.5.10.10 50 or 50 100 or 100 150 or 150-200 times compared to the corresponding compstatin analog having the same sequence (and containing one or more blocking moieties where appropriate) but not containing the specified GFR. It should be understood that, in various embodiments, such an increase in half-life may be observed following administration by other routes, such as subcutaneous administration, and/or other doses, such as other doses described herein, such as 20 μg/kg.

[00195] As noted, in some embodiments, the compstatin analog of any of SEQ ID NOs: 3-36, 37, 71, 72, 73, 74, or 75 is extended with one or more amino acids at the N-terminus, C-terminus, or at both ends, wherein at least one of said amino acids contains a side chain containing a reactive functional group such as a primary or secondary amine, a sulfhydryl group, a carboxyl group (which may be present as a carboxylate group), a guanidino group, a phenol group, an indole ring , thioether or imidazole ring, which facilitates conjugation with a reactive functional group with the addition of GFR to the specified compstatin analog. It should be understood that the corresponding non-GFR compstatin analog may also lack one or more of those amino acids present in the long-acting compstatin analog to which it corresponds. Thus, the corresponding compstatin analogue containing any of SEQ ID NOs: 3 36, 37, 71, 72, 73, 74, or 75 and not containing GFR is intended to "have the same amino acid sequence" as the sequences of SEQ ID NO: 3-36, 37, 71. 72, 73, 74 or 75, respectively. For example, a corresponding compstatin analog containing the amino acid sequence of SEQ ID NO: 14, 21, 28, 29, 32, 33, 34, or 36 and not containing GFR is meant to "have the same amino acid sequence" as SEQ ID NO : 14, 21, 28, 29, 32, 33, 34 or 36, respectively.

[00196] In some embodiments, the plasma half-life is the terminal half-life after a single intravenous dose. In some embodiments, the plasma half-life is the final half-life after reaching steady state after multiple intravenous doses. In some embodiments, after administration of a single IV dose to a primate, or after administration of multiple IV doses, a long-acting compstatin analog plasma Cmax value is achieved that is at least 5-fold higher than that of the corresponding GFR-free compstatin analog, e.g., 5-50 times. In some embodiments, after administration of a single IV dose to a primate, or after administration of multiple IV doses, a long-acting compstatin analog plasma Cmax value is 10-20 times higher than that of the corresponding GFR-free compstatin analog.

[00197] In some embodiments, the primate is a human. In some embodiments, the primate is a non-human primate, such as a monkey, such as a cynomolgus or rhesus monkey.

[00198] In some embodiments, the renal clearance of the long-acting compstatin analogue during the first 24 hours after 10 mg/kg or 20 μg/kg IV administration in humans or non-human primates is at least 2-fold lower, for example, 25, 510, 1050 or 50100 or 100150 or 150200 times the renal clearance of the corresponding compstatin analog. It should be understood that, in various embodiments, such a decrease in renal clearance may be observed following administration by other routes, such as subcutaneous administration, and/or other doses, such as other doses described herein, such as 20 μg/kg.

[00199] The concentration of the compstatin analogue in blood and/or urine samples can be measured using, for example, UV, HPLC, mass spectroscopy (MS), or anti-GFR antibodies, or a combination of methods such as LC/MS or LC/MS/ MS. Pharmacokinetic parameters such as half-life and clearance can be determined using methods known to those skilled in the art. Pharmacokinetic analysis can be performed, for example, using WinNonlin v 5.2 software (Pharsight Corporation, St. Louis, MO) or other suitable programs.

[00200] In certain embodiments, GFR is stable under physiological conditions for at least 24 hours or more. In certain embodiments, GFR is stable in the blood, plasma, or serum of a mammal, eg, a primate, eg, a human, or a non-human primate (eg, a monkey) for at least 24 hours. In various embodiments, at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the GFR molecules remain intact when incubated under physiological conditions for 24 hours, 48 hours, 72 hours, 96 hours, 120 hours, 144 hours, 168 hours or more. In various embodiments, at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the GFR molecules remain intact when incubated in blood, plasma or serum at 37 degrees C for 48 hours, 72 hours, 96 hours, 120 hours, 144 hours, 168 hours or more. Incubation can be carried out using GFR at a concentration of 1 μg/ml to about 100 mg/ml according to various implementation options. Samples can be analyzed at different time points. Size or intactness can be assessed using, for example, chromatography (eg, HPLC), mass spectrometry, Western blot, or any other suitable method. Such stability characteristics can be conferred on the GFR-conjugated fragment. In various embodiments, the GFR long-acting analog of compstatin may have any of the aforementioned stability characteristics. In some aspects, intact to the long acting compstatin analog means that said fragment of the compstatin analog remains conjugated to the GFR and the size of the GFR remains approximately the same as at the start of incubation or administration.

[00201] In some embodiments, the long acting compstatin analogue has a molar activity of at least about 10%, 20%, 30%, e.g., 30% to 40%, 30% to 50%, 30% to 60 %, 30% to 70%, 30% to 80%, 30% to 90% or more of the activity of the corresponding compstatin analogue having the same amino acid sequence (and, where appropriate, containing one or more blocking fragments), but not containing GFR. In some embodiments where the long acting compstatin analog comprises multiple compstatin analog moieties, said long acting compstatin analog has a molar activity of at least about 10%, 20%, or 30%, e.g., 30% to 40%, 30% up to 50%, from 30% to 60%, from 30% to 70%, from 30% to 80%, from 30% to 90% or more of the total activity of these compstatin analog fragments.

[00202] In some embodiments, the polyethylene glycol (PEG) contains a (CH ₂ CH ₂ O) _n moiety with a molecular weight of at least 500 daltons.

[00203] In some embodiments, the linker described above contains a (CH ₂ CH ₂ O) _n fragment with an average molecular weight of from about 500; 1000; 1500; 2000; 5000; 10,000; 20,000; 30,000; 40,000; 50,000; 60,000; 70,000; 80,000; 90,000; up to 100,000 daltons.

[00204] In some embodiments, the average molecular weight of the PEG is at least 20,000 Daltons, and up to about 100,000; 120,000; 140,000; 160,000; 180,000; or 200,000 daltons. "Average molecular weight" refers to the average molecular weight. In some embodiments, the polydispersity D of (CH ₂ CH ₂ O) _n is 1.0005 to 1.50, eg 1.005 to 1.10; 1.15;1.20; 1.25; 1.30; 1.40 or 1.50; or takes any value between 1.0005 and 1.50.

[00205] In some embodiments, the CH ₂ CH ₂ O) _n moiety is monodisperse and the polydispersity value of the (CH ₂ CH ₂ O) _n moiety is 1.0. Such CH ₂ CH ₂ O) _n monodisperse fragments are known in the art, commercially available from Quanta BioDesign (Powell, Ohio) and include, by way of non-limiting example, monodisperse fragments whose n value is 2, 4, 6, 8, 12, 16, 20 or 24.

[00206] In some embodiments, a compound contains multiple (i.e., multiple) (CH ₂ CH ₂ O) _n moieties, characterized in that the total molecular weight of said (CH ₂ CH ₂ O) _n moieties is between about 1000, 5000 , 10,000, 20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000 to 100,000 daltons. In some embodiments, the average total molecular weight of said compound or CH ₂ CH ₂ O) _n moieties is at least 20,000 Daltons, and up to about 100,000; 120,000; 140,000; 160,000; 180,000; or 200,000 daltons. In some embodiments, said compound contains multiple CH ₂ CH ₂ O) _n fragments of a specific length, e.g., n=4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 or more. In some embodiments, said compound contains a sufficient number of CH ₂ CH ₂ O) _n fragments of a certain length such that the total molecular weight of said CH ₂ CH ₂ O) _n fragments is from about 1000, 5000, 10,000, 20,000, 30,000, 40 000, 50,000, 60,000, 70,000, 80,000, 90,000 to 100,000 daltons. In some embodiments, the average total molecular weight of said compound or CH ₂ CH ₂ O) _n moieties is at least 20,000 Daltons and up to about 100,000; 120,000; 140,000; 160,000; 180,000; or 200,000 daltons. In some embodiments, the value of n is from about 30 to about 3000.

[00207] In some embodiments, a compstatin analog moiety is attached at each end of the linear PEG. A bifunctional PEG containing a reactive functional group at each end of said chain can be used, for example, as described above. In some embodiments, said reactive functional groups are identical, while in some embodiments, different reactive functional groups are present at each end.

[00208] In some embodiments, several CH ₂ CH ₂ O) _n moieties are represented as branched structures. The branches may be attached to a linear polymeric backbone (eg in a comb structure) or may extend from one or more groups of the central core, eg in a star structure. In some embodiments, the branched molecule contains 3 to 10 CH ₂ CH ₂ O) _n chains. In some embodiments, the branched molecule contains 4 to 8 chains (CH ₂ CH ₂ O) _n In some embodiments, the branched molecule contains 10, 9, 8, 7, 6, 5, 4, or 3 chains CH ₂ CH ₂ O) _n . In some embodiments, the star-shaped molecule contains 10-100, 10-50, 10-30, or 10-20 CH ₂ CH ₂ O) _n chains extending from the central core group. In some embodiments, the long-acting compstatin analog may thus contain, for example, 3-10 compstatin analog moieties, e.g., 4-8 compstatin analog moieties, each attached to the CH ₂ CH ₂ O) _n chain with a functional group at the end the specified chain. In some embodiments, the long acting compstatin analog may comprise, for example, 10-100 compstatin analog moieties, each attached to the CH ₂ CH ₂ O) _n chain with a functional group at the end of said chain. In some embodiments, the branches (sometimes referred to as "arms") of the branched or star-shaped PEGs contain approximately the same number of moieties (CH ₂ CH ₂ O). In some embodiments, the lengths of at least some of the branches may differ. It should be understood that, in some embodiments, one or more of the CH ₂ CH ₂ O) _n chains does not contain an attached compstatin analog moiety. In some embodiments, at least about 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% of said chains have a compstatin analog moiety attached.

[00209] In general, in the compounds shown herein, the polyethylene glycol moiety is depicted with an oxygen atom on the right side of the repeat unit or on the left side of the repeat unit. Where only one orientation is depicted, the present invention encompasses both orientations (ie, CH ₂ CH ₂ O) _n and CH ₂ CH ₂ O) _n of polyethylene glycol moieties for a particular compound or class; or, in cases where the compound or class contains multiple polyethylene glycol moieties, all orientation combinations are encompassed by the present invention.

[00210] The formulas of some examples of monofunctional PEGs containing a reactive functional group are given below. For illustrative purposes, formulas are shown in which said reactive functional group(s) includes an NHS ester, but other reactive functional groups, for example as described above, may be used. In some embodiments, (CH ₂ CH ₂ O) _n is shown with a methoxy group (OCH ₃ ) at the left end, however, it should be understood that the chains depicted below and elsewhere herein may end in a different OR moiety (e.g., an aliphatic group, an alkyl group, a lower alkyl group, or any other suitable PEG end group) or an OH group. It should also be borne in mind that, according to various implementation options fragment (CH ₂ CH ₂ O) _n NHS-group can connect fragments other than those shown.

[00211] In some embodiments, the monofunctional PEG has formula A:

where "Reactive functional group" and n are as defined above and the descriptions of classes and subclasses herein;

R ¹ represents hydrogen, an aliphatic group, or any suitable end group; and

T is a covalent bond or straight or branched C _1-12 hydrocarbon chain, where one or more T carbon units are optionally and independently replaced by -O-, -S-, -N(RX)-, -C(O)-, -C(O)O-, -OC(O)-, -N(R ^x )C(O)-, -C(O)N(R ^x )-, -S(O)-, -S(O ) ₂ -, -N(R ^x )SO ₂ - or -SO ₂ N(R ^x )-; and each R ^x is independently hydrogen or aliphatic C _1-6 .

[00212] Examples of monofunctional PEGs of Formula A include:

[00213] In formula I, the moiety containing the reactive functional group has the general structure -CO-(CH ₂ ) _m -COO-NHS, where m=2. In some embodiments, the monofunctional PEG has a structure corresponding to formula I, where m is 1 to 10, such as 1 to 5. For example, in some embodiments, m is 3, as shown below:

[00214] In formula II, the moiety containing the reactive functional group has the general structure -(CH ₂ ) _m -COO-NHS, where m=1. In some embodiments, the monofunctional PEG has a formula II structure where m is from 1 to 10 (e.g., where m is 5 as shown in Formula III below), or where m is 0 (as shown in Formula below). Sha).

[00215] In some embodiments, a bifunctional linear PEG contains a fragment containing a reactive functional group at each end. Said reactive functional groups may be the same (homobifunctional) or different (heterobifunctional). In some embodiments, the structure of the bifunctional PEG may be symmetrical, with identical moieties used to link the reactive functional group to the oxygen atoms at each end of the -(CH ₂ CH ₂ O) _n chain. In some embodiments, different moieties are used to link said two reactive functional groups to the PEG moiety of said molecule. The structures of exemplary bifunctional PEGs are shown below. For illustrative purposes, formulas are given in which the specified(s) reactive(s) functional group(s) include an NHS ester, but other reactive functional groups can be used.

[00216] In some embodiments, the bifunctional linear PEG has formula B:

where each T and "Reactive functional group" independently correspond to the above definitions and are described in classes and subclasses in this document, and n corresponds to the above definitions and is described in classes and subclasses in this document.

[00217] Examples of bifunctional PEGs having Formula B include:

[00218] In formula IV, the moiety containing the reactive functional group has the general structure -(CH ₂ ) _m -COO-NHS, where m=1. In some embodiments, the bifunctional PEG has a structure corresponding to formula IV, where the value of m is from 1 to 10, for example, from 1 to 5. In certain embodiments, m=0, for example, a moiety containing a reactive functional group has the general structure - COO-NHS. For example, in some embodiments, the bifunctional PEG has the structure of formula IVa as shown below:

[00219] In formula V, the moiety containing the reactive functional group has the general structure -CO-(CH ₂ ) _m -COO-NHS, where m=2. In some embodiments, bifunctional PEGs have a structure corresponding to formula V, where the value of m is from 1 to 10, for example, from 1 to 5. In certain embodiments, for example, m=2, as shown below:

[00220] In some embodiments, the present invention provides a polymer-conjugated analogue of compstatin. In some embodiments, to synthesize such conjugates, a functional group (eg, an amine, hydroxyl, or thiol group) is reacted on a compstatin analog with a PEG-containing compound having a "reactive functional group" as described herein. For example, formulas III and IV, respectively, can form compstatin analog conjugates with the structure:

на аналоге компстатина. Согласно определенным вариантам реализации аминогруппа представляет собой группу боковой цепи лизина.

analogous to compstatin. In certain embodiments, the amino group is a lysine side chain group.

[00221] It should be borne in mind that appropriate conjugates can be obtained with any of the PEG-containing compounds and classes described in this application, depending on the choice of the reactive functional group and/or the functional group of the compstatin. For example, formulas IVa and Va, respectively, can form compstatin analog conjugates with the structure

In certain embodiments, the PEG component of such conjugates has an average molecular weight of about 20 kDa-100 kDa, about 20 kDa-90 kDa, about 20 kDa-80 kDa, about 20 kDa-70 kDa, about 20 kDa-60 kDa, approximately 20 kDa-50 kDa, approximately 30 kDa-80 kDa, approximately 30 kDa-70 kDa, approximately 30 kDa 60 kDa, approximately 30 kDa 50 kDa, approximately 30 kDa 45 kDa, approximately 35 kDa 50 kDa, approximately 35 kDa 45 kDa , approximately 36 kDa-44 kDa, approximately 37 kDa-43 kDa, approximately 38 kDa-42 kDa or approximately 39 kDa-41 kDa. In certain embodiments, the PEG component of such conjugates has an average molecular weight of approximately 40 kDa.

[00222] The term "bifunctional" or "bifunctionalized" is sometimes used herein to refer to a compound containing two compstatin analog fragments associated with GFR. Such connections may be designated by the letters "BF". In some embodiments, the bifunctionalized compound is symmetrical. In some embodiments, the associations between the GFR and each of the compstatin analogue fragments of the bifunctionalized compound are identical. In some embodiments, each linkage between GFR and the compstatin analogue of the bifunctionalized compound contains a carbamate. In some embodiments, each linkage between GFR and the compstatin analog of the bifunctionalized compound contains a carbamate and does not contain an ester. In some embodiments, each compstatin analogue of the bifunctionalized compound is directly linked to GFR through a carbamate. In some embodiments, each compstatin analogue of the bifunctionalized compound is directly linked to GFR via a carbamate, and said bifunctionalized compound has the structure:

представляет точку присоединения группы боковой цепи лизина в аналоге компстатина, имеющем структуру:[00223] According to some embodiments of the formulas and embodiments described herein,

represents the point of attachment of the lysine side chain group in the compstatin analogue having the structure:

where the symbol indicates the point of attachment of the chemical fragment to the rest of the molecule or chemical formula.

[00224] In some embodiments, the branched, comb, or star-shaped PEG contains a fragment containing a reactive functional group at the end of each of several chains -(CH ₂ CH ₂ O) _n . These reactive functional groups may be the same, or at least two groups may be different. In some embodiments, the branched, comb, or star PEG has a formula selected from the following:

[00225] where each R2 is independently a "reactive functional group", or R ¹ , and each of T, n and "reactive functional groups" independently corresponds to the above definitions and is described in classes and subclasses in this application. The structure of examples of branched PEGs (with 8 arms or branches) containing NHS moieties as reactive functional groups is shown below:

[00226] The structure of exemplary branched PEGs (with 4 arms or branches) containing NHS fragments as reactive functional groups is depicted below:

[00227] The number of branches extending from the core may vary. For example, the number 4 in the above formulas VI and VII can be changed to any other integer from 0 to 10 according to various implementation options. In certain embodiments, one or more branches do not contain a reactive functional group; said branch ends with a -CH ₂ CH ₂ OH or -CH ₂ CH ₂ OR group, as described above.

[00228] In some embodiments, a branched PEG has a structure corresponding to formula VII, VIII, or IX (or variants thereof with a different number of branches), provided that x is

[00229] In some embodiments, a branched PEG has a structure corresponding to formula VII, VIII, or IX (or variants thereof with a different number of branches), provided that x is

[00230] Of course, the methylene (CH ₂ ) group in the above fragment x may alternatively contain a longer alkyl chain (CH ₂ ) _m , where m takes values up to 2, 3, 4, 5, 6, 8, 10, 20 or 30, or may contain one or more of the other fragments described in this application.

[00231] In some embodiments, examples of branched PEGs containing NHS or maleimide reactive groups are depicted below:

[00232] In some embodiments, a formula X or XI variant is used, where 3 or each of the 4 branches contains a reactive functional group.

[00233] The following examples of PEGs may also be given:

[00234]

As already noted, it should be borne in mind that, as described in this application, in various implementations, any of a variety of fragments can be introduced between the peptide component and the (CH ₂ CH ₂ O) _n -R fragment of the long-acting analog of compstatin, such as linear alkyl, ester, amide, aromatic ring (eg, substituted or unsubstituted phenyl), substituted or unsubstituted cycloalkyl structure, or combinations thereof. In some embodiments, such(s) fragments) may render the compound more susceptible to hydrolysis, which may detach the peptide portion of said compound from the GFR. In some embodiments, such release may increase in vivo tissue penetration and/or increase the activity of said compound. In some embodiments, the hydrolysis is total (eg, acid-base) hydrolysis. In some embodiments, the hydrolysis is enzyme-catalyzed, eg, esterase-catalyzed. Of course, both types of hydrolysis can occur. Examples of PEGs containing one or more of these moieties and an NHS ester as a reactive functional group are given below:

[00235]

[00236]

[00237]

[00238] In some embodiments, the branched (multiple branches) PEG or star PEG contains a pentaerythritol core, a hexaglycerol core, or a tripentaerythritol core. It should be understood that, in some embodiments, the branches do not necessarily radiate from the same point.

[00239] Monofunctional, bifunctional, branched, and other PEGs containing one or more reactive functional groups can, in some embodiments, be obtained from, for example, NOF America Corp. White Plains, NY or BOC Sciences, 45-16 Ramsey Road, Shirley, New York 11967, USA, including, or can be obtained using methods known in the art.

[00240] In some embodiments, the bond between GFR and the compstatin analog contains a carbamate. In some embodiments, the compstatin analog is directly linked to GFR via a carbamate. In some embodiments, the bond between GFR and the compstatin analog does not contain an ester. In some embodiments, the linkage between GFR and the compstatin analog contains a carbamate and does not contain an ester. In some embodiments, the bond between GFR and the compstatin analog contains a carbamate and does not contain a bond that is more susceptible to hydrolysis in an aqueous environment than carbamate. In some embodiments, the SCF contains a PEG moiety or consists of a PEG moiety.

[00241] In some embodiments, the linkage between the GFR and the compstatin analog contains an amide. In some embodiments, the compstatin analogue is directly linked to GFR via an amide. In some embodiments, the linkage between GFR and the compstatin analog contains an amide and does not contain an ester. In some embodiments, the bond between GFR and the compstatin analog contains an amide and does not contain a bond that is more susceptible to hydrolysis in an aqueous environment than an amide. In some embodiments, the SCF contains a PEG moiety or consists of a PEG moiety.

[00242] In some embodiments, one or more compstatin analogs of a multifunctionalized compound (eg, a bifunctionalized, trifunctionalized, or more highly functionalized compound) is linked to the GFR by a bond containing a carbamate. In some embodiments, one or more compstatin analogues of a multifunctional compound (eg, a bifunctional, trifunctional, or more highly functional compound) is linked to a GFR by a bond that does not contain an ester. In some embodiments, one or more compstatin analogs of a multifunctional compound (eg, a bifunctional, trifunctional, or more highly functional compound) is linked to a GFR by a bond that contains a carbamate and no ester. In some embodiments, one or more compstatin analogs of a multifunctional compound (e.g., a bifunctional, trifunctional, or more highly functional compound) is linked to a GFR bond that contains a carbamate and does not contain a bond that is more sensitive to aqueous hydrolysis than carbamate. In some embodiments, each compstatin analog of a multifunctional compound (eg, bifunctional, trifunctional, or higher functional compound) is directly linked to GFR via a carbamate.

[00243] In some embodiments, the SCF contains a PEG moiety or consists of a PEG moiety. In some embodiments, one or more compstatin analogs of a multifunctional compound (eg, a bifunctional, trifunctional, or more highly functional compound) is linked to the GFR by an amide-containing bond. In some embodiments, one or more compstatin analogs of a multifunctional compound (eg, a bifunctional, trifunctional, or more highly functional compound) is linked to a GFR by a bond that contains an amide and no ester. In some embodiments, one or more compstatin analogues of a multifunctional compound (e.g., a bifunctional, trifunctional, or higher functional compound) is linked to a GFR bond that contains an amide and does not contain a bond that is more susceptible to hydrolysis in aqueous media than the amide. In some embodiments, each compstatin analog of a multifunctional compound (eg, bifunctional, trifunctional, or higher functional compound) is directly linked to GFR via an amide. In some embodiments, the SCF contains a PEG moiety or consists of a PEG moiety.

[00244] In some embodiments, the present invention provides a compstatin analogue conjugated to a polymer, said polymer being non-PEG. In some embodiments, the polymer is a polyoxazoline (POZ). Examples of mono- and polyfunctionalized polyoxazoline derivatives for direct conjugation or for conjugation with a linker are shown below:

ZT-[N(COR ^x )CH ₂ CH ₂ ] _n -TR ¹ ;

R ¹ -{[N(CO-TZ)CH ₂ CH ₂ ] _m -[N(COR ^x )CH ₂ CH ₂ ] _n } ^a -TR ¹ ;

R ¹ -{[N(CO-TZ ¹ )CH ₂ CH ₂ ] _p -[N(COR ^x )CH ₂ CH ₂ ] _n -[N(CO-TZ ² )CH ₂ CH ₂ ] _m } ^a -TR ¹ ;

R ¹ -{[N(CO-TZ ¹ )CH ₂ CH ₂ ] _p -[N(COR ^x )CH ₂ CH ₂ ] _n -[N(CO-TZ ² )CH ₂ CH ₂ ] _m } ^a -TZ ;

R ¹ -[N(COR ^x )CH ₂ CH ₂ ] _n -TB(-R ¹ )(-TZ)-T-[N(COR ^x )CH ₂ CH ₂ ] _m -R ¹ ;

where:

each of Z, Z ¹ and Z ² independently represents a reactive functional group, corresponding to the above definition and described in classes and subclasses in this document;

each of T, R ^x and R ¹ independently corresponds to the above definition and is described in classes and subclasses in this document;

each of m, n, and p is independently an integer from 0 to 1000, with the limitation that the sum, p, for each formula is not 0;

a="ran", i.e. random ("random") copolymer, or "block", i.e. block copolymer;

B is a branched moiety linked, with or without a linker, to other parts of the polymer.

Other examples of functionalized polyoxazoline derivatives for conjugation are described in detail in the art, including but not limited to those described in PCT Published Patent Applications Nos. , each of which is incorporated herein by reference in its entirety.

[00245] Examples of compstatin analog conjugates with polyoxazoline polymers are shown below:

where each variable independently corresponds to the above definition and is described in classes and subclasses in this document.

[00246] In some embodiments, the present invention provides a polymer-conjugated analog of compstatin, wherein said analog of compstatin is linked to the polymer via one or more linkers. In some embodiments, the polymer is selected from the PEG-containing compounds and genera described above and in the classes and subclasses herein. In some embodiments, the present invention provides conjugates of compstatin analogues to the PEG-containing compounds and genera described herein, characterized in that said compstatin analogue is connected to the PEG-containing moieties via one or more linkers. Mono- and polyfunctional PEGs that contain one or more reactive functional groups for conjugation are defined above and are described in classes and subclasses herein, including but not limited to the corresponding formulas A, I, Ia, II, III, IIIa, B, IV, IVa, V, Va, C, D, E, F, G, H, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, or XVI.

[00247] Suitable linkers for connecting a compstatin analog and a polymer moiety, such as PEG or polyoxazoline, are detailed above and in the classes and subclasses herein. In some embodiments, the linker contains multiple functional groups, with one functional group linked to the compstatin analog and the other linked to the polymer moiety. In some embodiments, the linker is a bifunctional compound. In some embodiments, the linker has the structure NH ₂ (CH ₂ CH ₂ O) _n CH ₂ C(=O)OH, where n is from 1 to 1000. In some embodiments, the linker is 8-amino-3,6- dioxaoctanoic acid (AEEAc). In some embodiments, the linker is activated to conjugate to a polymer moiety or a functional group of a compstatin analogue. For example, in some embodiments, the AEEAc carboxyl group is activated prior to conjugation to the side chain amino group of the lysine group.

[00248] In some embodiments, conjugation to the polymer moiety, either directly or via a linker, uses an appropriate functional group (eg, amine, hydroxyl, thiol, or carboxylic acid group) on the compstatin analog. In some embodiments, the compstatin analog is conjugated via an amine group to a PEG moiety by a linker. In some embodiments, the amine group is an a-amino group of an amino acid residue. In some embodiments, the amine group is a lysine side chain amine group. In some embodiments, the compstatin analog is conjugated to the PEG moiety via a lysine side chain amino group (s-amino group) with a linker having the structure NH ₂ (CH ₂ CH ₂ O) _n CH ₂ C(=O)OH, where n is from 1 to 1000. In some embodiments, the compstatin analog is conjugated to the PEG moiety via the amino group of the lysine side chain with an AEEAc linker. In some embodiments, the NH ₂ (CH ₂ CH ₂ O) _n CH ₂ C(=O)OH linker introduces the NH(CH ₂ CH ₂ O) _n CH ₂ C(=O) moiety onto the lysine side chain of compstatin after conjugation. In some embodiments, the AEEAc linker introduces a -NH(CH ₂ CH ₂ O) ₂ CH ₂ C(=O)- moiety onto the lysine side chain of compstatin after conjugation.

[00249] In some embodiments, the compstatin analog is conjugated to the polymer moiety by a linker, said linker comprising an AEEAc moiety and an amino acid residue. In some embodiments, the compstatin analog is conjugated to the polymer moiety by a linker, said linker containing an AEEAc moiety and a lysine residue. In some embodiments, the polymer is PEG. In some embodiments, the C-terminus of the compstatin analog is connected to an AEEAc amino group, and the C-terminus of AEEAc is connected to a lysine residue. In some embodiments, the C-terminus of the compstatin analog is connected to the amino group of AEEAc, and the C-terminus of AEEAc is connected to the a-amino group of the lysine residue. In some embodiments, the C-terminus of the compstatin analog is connected to the amino group of AEEAc, the C-terminus of AEEAc is connected to the a-amino group of the lysine residue, and the polymer moiety, e.g., the PEG moiety, is conjugated through the s-amino group of said lysine residue. In some embodiments, the C-terminus of the lysine residue is modified. In some embodiments, the C-terminus of the lysine residue is modified by amidation. In some embodiments, the N-terminus of the compstatin analog is modified. In some embodiments, the N-terminus of the compstatin analog is acetylated.

эбозначает точку присоединения аминной группы на аналоге компстатина,

обозначает аналог компстатина, присоединенный через С-конец, и при этом каждая из других переменных независимым образом соответствует приведенному выше определению и описана в классах и подклассах в настоящем документе. Согласно некоторым вариантам реализации аминная группа представляет собой аминогруппу боковой цепи лизина.[00250] Examples of conjugates containing an AEEAc linker and a polymer are shown below, with

e denotes the point of attachment of the amine group on the compstatin analogue,

denotes a compstatin analogue attached through the C-terminus, and while each of the other variables independently corresponds to the above definition and is described in classes and subclasses in this document. In some embodiments, the amine group is a lysine side chain amino group.

[00251] In certain embodiments, a compstatin analogue can be represented as M-AEEAc-Lys-B ₂ where B ₂ is a blocking moiety, e.g., NH ₂ , M is any of SEQ ID NOs: 3-36, 37, 71, 72, 73, 74, or 75, provided that the C-terminal amino acid corresponding to any of SEQ ID NOs: 3-36, 37, 71, 72, 73, 74, or 75 is peptide-linked to AEEAc-Lys- The B ₂ NHS moiety of a monofunctional or multifunctional (e.g. bifunctional) PEG reacts with the free amine of the lysine side chain to form a monofunctionalized (one compstatin analog moiety) or multifunctionalized (multiple compstatin analog moieties) long acting compstatin analog. In various embodiments, any amino acid containing a side chain that contains a reactive functional group can be used instead of Lys (or in addition to Lys). A monofunctional or multifunctional PEG containing an appropriate reactive functional group can be reacted with such a side chain in a similar manner to the reaction of NHS-activated PEG with Lys.

[00252] With respect to any of the formulas and structures described above, it should be understood that embodiments in which the compstatin analog component comprises any compstatin analog described herein, for example, any compstatin analog corresponding to the sequences of SEQ ID NOs; 3-36, 37, 71, 72, 73, 74, 75. For example, without limitation, the compstatin analog may comprise an amino acid sequence corresponding to the sequence of SEQ ID NO: 28. An example of a long-acting compstatin analog wherein the compstatin analog component contains the sequence amino acids corresponding to the sequence of SEQ ID NO: 28 is depicted in FIG. 10(C). It should be understood that a PEG fragment may have a different molecular weight or average molecular weight in different embodiments, as described herein. For example, the molecular weight of the individual chains in a formulation may vary and/or different formulations may have different average molecular weight and/or polydispersity as described herein. In certain embodiments, the PEG moiety in the compound of FIG. 10(C) has an average molecular weight of approximately 20 kDa to 100 kDa, approximately 20 kDa-90 kDa, approximately 20 kDa-80 kDa, approximately 20 kDa-70 kDa, approximately 20 kDa-60 kDa, approximately 20 kDa- 50 kDa, approximately 30 kDa-80 kDa, approximately 30 kDa-70 kDa, approximately 30 kDa-60 kDa, approximately 30 kDa-50 kDa, approximately 30 kDa-45 kDa, approximately 35 kDa-50 kDa, approximately 35 kDa-45 kDa, approximately 36 kDa-44 kDa, approximately 37 kDa-43 kDa, approximately 38 kDa-42 kDa, or approximately 39 kDa-41 kDa. In some embodiments, the PEG moiety in the compound of FIG. 10(C) has an average molecular weight of about 30 kD to about 50 kD, eg, about 35 kD to about 45 kD, about 37.5 kD to about 42.5 kD. In certain embodiments in which the PEG moiety has an average molecular weight of approximately 40 kDa, e.g., 37.5 kDa to 42.5 kDa, 38 kDa, 39 kDa, 40 kDa, 41 kDa, 42 kDa, referred to in this document as CA28-2TS-BF. In certain embodiments, a GFR-containing compound, e.g., a PEG moiety that has an average molecular weight of approximately 40 kDa, e.g., 37.5 kDa to 42.5 kDa, 38 kDa, 39 kDa, 40 kDa, 41 kDa, 42 kDa, the terminal half-life of said compound is at least about 5 days, e.g., about 5-10 days, e.g., about 5, 6, 7, 8, 9 days, when administered intravenously or subcutaneously to non-human primates or humans, for example, at a dose of about 1-3 µg/kg, 3-5 µg/kg, or 5-10 µg/kg.

[00253] In some aspects, the present invention relates to the use of click chemistry with compstatin analogues. "Click chemistry" is well known in the art and is suitable for use in some aspects of the present invention. Click chemistry includes, in some embodiments, versatile cycloaddition reactions between azides and alkynes, providing a number of useful applications. Methods for carrying out click chemistry reactions are known in the art and are described in the sources: Kolb, H.C.; Sharpless, K. V., Drug Disc. Today, 2003, 1128-1137; Moses, J.E.; Moorhouse, A. D.; Chem. soc. Rev., 2007, 1249-1262; which are incorporated herein by reference in their entirety. Click chemistry is a common bioconjugation technique due to its high reactivity and selectivity, even in biological media. See: Kolb, H.C.; Finn, M. G.; Sharpless, K.V. Angew. Chem. Int. Ed. 2001, 40, 2004-2021; and Wang, Q.; Chan, T.R.; Hilgraf, R.; Fokin, V. V.; Sharpless, K. V.; Finn, M. G. J. Am. Chem. soc. 2003, 125, 3192-3193. In addition, currently available recombinant techniques and synthesis methods allow the introduction of azides and alkyne-bearing non-canonical amino acids into peptides, proteins, cells, viruses, bacteria and other biological objects that consist of proteins or display proteins. See: Link, A.J.; Vink, M. K. S.; Tirrell, D. A. J. Am. Chem. soc. 2004, 126, 10598-10602; Deiters, A.; Cropp, T. A.; Mukherji, M.; Chin, J. W.; Anderson, C; Schultz, P. G. J. Am. Chem. soc. 2003, 125, 11782-11783.

[00254] As used herein, the term "click chemistry" is sometimes used to refer to a reactive functional group capable of participating in a click chemistry reaction with a suitable second reactive functionality, said second reactive functionality also being a click chemistry group. . Said first and second click chemical groups or entities (eg molecules) containing such groups may be referred to as complementary. The first and second entities, such as molecules that contain complementary click chemical groups, may be referred to as click chemical partners. An object or molecule containing a click-chemical group may be referred to as "click-functionalized". A bond formed by the reaction of complementary click chemical partners may be referred to as a "click chemical bond".

[00255] In some embodiments, the present invention provides click-functionalized analogs of compstatin, for example, for conjugation to a complementary moiety on a partner molecule or partner biomolecule. In some embodiments, the complementary partner molecule or biomolecule partner is a polymer, peptide, protein, or molecule that functions as a clearance-reducing moiety. In some embodiments, said "click-functionalized" moiety is an alkyne or an alkyne derivative capable of undergoing [3+2] cycloaddition reactions with complementary azide-bearing molecules and biomolecules. In another embodiment, the "click-functionalized" functional group is an azide or azide derivative capable of undergoing [3+2] cycloaddition reactions with complementary alkyne-bearing molecules and biomolecules (i.e., click chemistry).

[00256] In some embodiments, the click-functionalized compstatin analog bears an azide group on any side chain group of the compstatin analog. In some embodiments, the click-functionalized analog of compstatin carries an azide group on a lysine side chain group.

[00257] In some embodiments, the click-functionalized compstatin analog bears an alkyne group on any side chain group of the compstatin analog. In some embodiments, the click-functionalized analog of compstatin carries an alkyne group on a lysine side chain group.

[00258] In some embodiments, the present invention provides compstatin conjugates comprising a compstatin analog, a molecule that functions as a clearance-reducing moiety, and a triazole linker. In some embodiments, the triazole linker is the result of a click-chemical conjugation between a compstatin conjugate and a molecule that functions as a clearance-reducing moiety. In some embodiments, the FFR may be any FFR disclosed herein. For example, the GFR may be a PEG, a polypeptide, or a POS.

[00259] In some embodiments, the present invention provides compstatin conjugates comprising a compstatin analog, a PEG moiety, and a triazole linker. In some embodiments, the triazole linker is the result of a click-chemical conjugation between the compstatin conjugate and the PEG moiety.

[00260] In some embodiments, the present invention provides compstatin conjugates comprising a compstatin analog, a polyoxazoline moiety, and a triazole linker. In some embodiments, the triazole linker is the result of a click-chemical conjugation between a compstatin conjugate and a polyoxazoline moiety.

[00261] In some embodiments, the click chemistry reaction between the compstatin analog and another moiety is catalyzed by a transition metal. Copper-containing molecules that catalyze the "click" reaction include, but are not limited to, copper wire, copper bromide (CuBr), copper chloride (CuCl), copper sulfate (CuSO ₄ ), copper sulfate pentahydrate (CuSO ₄ ⋅5H ₂ O ), copper acetate (Cu ₂ (AcO ₄ ), copper iodide (CuI), [Cu(MeCN) ₄ ](OTf), [Cu(MeCN) ₄ ](PF ₆ ), colloidal copper sources and immobilized copper sources. According to in some embodiments, other metals such as ruthenium Reducing agents and organic and inorganic metal binding ligands can be used in combination with metal catalysts and include, but are not limited to, sodium ascorbate, tris(triazolyl)amine ligands, tris(carboxyethyl) phosphine (TCEP), sulfonated bathophenanthroline ligands and benzimidazole based ligands.

[00262] In some embodiments, compstatin analogs are conjugated to other moieties using metal-free click chemistry (also known as copper-free click chemistry) to provide a metal-free composition or conjugates. Unlike standard click chemistry, also known as copper-catalyzed click chemistry (CuACC), metal-free click chemistry reactions take place between either a deformed cyclic alkyne or an alkyne precursor such as oxanorbornadiene and an azide group. As the name suggests, a metal catalyst is not necessary for the reaction to proceed. Examples of such chemical reactions include those involving cyclooctyne derivatives (Codelli, et. al., J. Am. C hem. Soc, 2008, 130, 11486-11493; Jewett, et. al., J. Am. Chem. Soc, 2010, 132, 3688-3690; Ning, et. al., Angew. Chem. Int. Ed., 2008, 47, 2253-2255), difluoro-oxanorbornene derivatives (van Berkel, et. al., ChemBioChem, 2007, 8, 1504-1508) or nitrile oxide derivatives (Lutz, et. al., Macromolecules, 2009, 42, 5411-5413). In certain embodiments, the metal-free click reaction is a [3+2] cycloaddition reaction, a Diels-Alder reaction, or a metal-free radical thiol-alkene addition reaction. Examples of click chemistry reactions and click chemistry groups are described in, for example, Joerg Lahann, Click Chemistry for Biotechnology and Materials Science, 2009, John Wiley & Sons Ltd, ISBN 978-0-470-69970-6; Becer, Hoogenboom, and Schubert, Click Chemistry beyond Metal-Catalyzed Cycloaddition, Angewandte Chemie International Edition (2009) 48: 4900 4908. In certain embodiments, the click chemistry group contains diarylcyclooctyne.

[00263] Some examples of click chemical reactions without metals are shown in the scheme below.

(по Ning et. al; Angew Chem Lit Ed, 2008, 47, 2253); двуфтористые циклооктины (DIFO или DFO)

(no Codelli, et. al.; J. Am. Chem. Soc. 2008, 130, 11486-11493.); биарилазациклооктинон (BARAC)

(no Jewett et. al.; J. Am. Chem. Soc. 2010, 132, 3688); или бициклононин (BCN)

(по Dotnmerholt, et al.; Angew Chem bit Ed, 2010, 49, 9422-9425) или дибензилциклооктин (DBCO)

[00264] Certain click fragments without metals are known from the literature. Examples include 4-dibenzocyclooctynol (DIBO)

(after Ning et. al; Angew Chem Lit Ed, 2008, 47, 2253); difluoric cyclooctynes (DIFO or DFO)

(no Codelli, et. al.; J. Am. Chem. Soc. 2008, 130, 11486-11493.); biarylazacyclooctinone (BARAC)

(no Jewett et. al.; J. Am. Chem. Soc. 2010, 132, 3688); or bicyclononin (BCN)

(according to Dotnmerholt, et al.; Angew Chembit Ed, 2010, 49, 9422-9425) or dibenzylcyclooctyne (DBCO)

[00265] A reaction scheme involving the reaction of DBCO and azide is shown below:

[00266] In the above scheme, according to various embodiments, A may contain or consist of a compstatin analog fragment, and B may contain or consist of GFR, for example, a polymer such as PEG or POS, or a polypeptide, or B may contain or consist of from a fragment of a compstatin analogue and may contain or consist of a GFR, for example, a polymer such as PEG or POS, or a polypeptide.

[00267] In some embodiments, the "click-functionalized metal-free moiety" is acetylene or an acetylene derivative capable of undergoing [3+2] cycloaddition reactions with complementary azide-bearing molecules and biomolecules without the use of a metal catalyst.

[00263] In some embodiments, the R and R' groups of the metal-free click chemicals are a compstatin analog or any molecule described herein to which a compstatin analog can be conjugated. In some embodiments, such compstatin analogs carry a click-functionalized moiety on the lysine side chain. In some embodiments, such compstatin analogs are linked to the click-functionalized moiety by a linker. In some embodiments, such compstatin analogs are coupled to the click-functionalized moiety via AEEAc.

[00269] In some embodiments, the click chemical reagent contains DBCO. Examples of reagents and examples of their use are given below:

[00270] DBCO acid. In some embodiments, the DBCO acid may be used to react with the amine containing moiety.

[00271] A DBCO-NHS ester (above) or a BBCO-sulfo-NHS ester (below) can be used to incorporate a DBCO functional group into an amine-containing molecule, such as a compstatin analogue or a polypeptide containing a lysine residue.

[00272] DBCO-PEG4-NHS-Ether. In some embodiments, such a reagent is suitable for introducing a DBCO moiety via reaction with an available amine functional group. In some aspects, the presence of a PEG chain as a hydrophilic spacer may be useful for, for example, increasing solubility or providing flexibility.

[00273] DBCO-amine. In some embodiments, the click chemical reagent contains a carbonyl/carboxyl-reactive dibenzylcyclooctyne that can react with acids, reactive esters, and/or aldehydes.

[00274] In certain embodiments, the click chemistry reaction includes the cyclooctyne depicted below:

[00275] In certain embodiments, click chemistry reactions include reactions between nitrones and cyclooctynes (see, for example, Ning, Xinghai; Temming, Rinske P.; Dommerholt, Jan; Guo, Jun; Ania, Daniel B.; Debets, Marjoke F.; Wolfert, Margreet A.; Boons, Geert-Jan et al. (2010) "Protein Modification by Strain-Promoted Alkyne-Nitrone Cycloaddition" Angewandte Chemie International Edition 49 (17): 3065), oxime/hydrazone formation from aldehydes and ketones, ligation of tetrazines (see, for example, Blackman, Melissa L.; Royzen, Maksim; Fox, Joseph M. (2008). "The Tetrazine Ligation: Fast Bioconjugation based on Inverse-electron-demand Diels-Alder Reactivity Journal of the American Chemical Society 130 (41): 13518-9), tetrazole ligation, isonitrile-based click reaction (see e.g. Stackmann, Henning; Neves, André A.; Stairs, Shaun; Brindle, Kevin M., Leeper, Finian J. (2011), "Exploring isonitrile-based click chemistry for ligation with biomolecules", Organic & Biomolecular Chemistry 9 (21): 7303) and quadricyclane ligation (see, for example, Sletten, Ellen M.; Bertozzi, Carolyn R. (2011). "A Bioorthogonal Quadricyclane Ligation". Journal of the American Chemical Society 133 (44): 17570-3). In certain embodiments, the click chemistry is Staudinger ligation (phosphine azide).

[00276] Any analogue of compstatin can be modified to include a click chemistry group according to various embodiments. For example, a compstatin analog containing a sequence corresponding to any of SEQ ID NOs: 3-36, 37, 71, 72, 73, 74, or 75 can be modified in this way. In some embodiments, any such sequence also contains a lysine residue or an AEEAc-Lys fragment, for example, at the C-terminus. In some embodiments, the click chemistry group is inserted after peptide synthesis. For example, the Lys side chain can be reacted with azidoacetic acid to introduce an azide moiety as the click chemistry group. In some embodiments, the click chemistry group is inserted after cyclization, and in some embodiments, after the addition of a blocking moiety at the N- and/or C-terminus. In some embodiments, the click chemistry group is inserted during peptide synthesis. For example, in the synthesis of a compstatin analog, an amino acid containing a side chain that contains a click chemical group can be used. Various such amino acids are commercially available from a number of sources, eg AAPPTes (Louisville, Kentucky), Jena Bioscience GmBH (Jena, Germany). In some aspects, provided herein are methods for preparing a click-chemically functionalized analog of compstatin.

[00277] In some embodiments, compositions are provided comprising a compstatin analogue and a click chemical. a click chemical group, for example, any click chemical group known in the art. In some aspects, the composition may be incubated under suitable conditions (which may include providing a suitable catalyst, light (eg, UV)) to functionalize the compstatin analog with the click chemical functional group. In some embodiments, the present invention provides analogs of compstatin that contain any click chemical group, including but not limited to those described herein. In some embodiments, methods are provided for making a long acting compstatin analogue. In some embodiments, these methods include mixing a compstatin analog containing a first click chemistry group with an SCF containing a complementary click chemistry group under conditions suitable for the click chemistry reaction to occur. Additional steps may include purification of the resulting conjugate. In some embodiments, purification comprises removing at least some of the unreacted components, for example, using a suitable scavenger.

[00278] In some embodiments, a click chemistry reaction is used to connect two or more SKFs, at least two of which contain an attached compstatin analogue moiety. Said compstatin analog fragments may be the same or different according to various embodiments. These fragments of the compstatin analog may or may not be attached to the GFR via a click chemical reaction. For example, in some embodiments, a first heterobifunctional PEG containing a first click moiety at the first end and an NHS ester at the second end is coupled to the compstatin analog moiety via an NHS ester. In a separate reaction, a second heterobifunctional PEG containing a second click chemistry group at the first end and an NHS ester at the second end is coupled to the compstatin analog moiety via an NHS ester. The resulting two compounds are then click-chemically reacted to produce a larger molecule containing two compstatin analogue moieties. PEG is mentioned as an example of GFR, however, it should be understood that this approach can be used with any GFR. For example, in some embodiments, it may be used with a GFR containing a polypeptide, such as HSA or a portion thereof, or an albumin or an albumin-binding peptide, or an antibody or portion thereof. In some embodiments, this approach may be applied with the POS.

[00279] Compstatin analogs containing a click chemistry group can be used in a variety of ways. In some embodiments, the compstatin analogue containing the first click chemical group is reacted with any substance containing the complementary click chemical group. A substance containing a complementary click chemical group may contain, for example, a label (eg, a fluorophore, a fluorescent protein, a radioactive isotope, etc.), an affinity reagent, an antibody, a targeting moiety, a metal, a particle, and the like. In some embodiments, a click chemistry group is used to attach a compstatin analog moiety to a surface, said surface containing or functionalized to contain a complementary click chemistry group. In some embodiments, the surface is sensory, such as a C3 capture/detection surface or sensor. In some embodiments, the surface forms part of a medical device, tubing, membrane, reservoir, implant, or other material that can contact blood (e.g., in vitro) or be temporarily or indefinitely implanted in a subject (e.g., a prosthesis or device for drug delivery). In some embodiments, the surface is functionalized with a compstatin analog to reduce complement activation thereon. In some embodiments, the device or tubing is used to circulate blood, such as for dialysis, during surgery, and the like. In some embodiments, the device is a hemodialyzer or an extracorporeal circulatory apparatus. Such compstatin analogue-functionalized devices and methods for their preparation are provided by the present invention.

[00280] According to some embodiments of the present invention, the compstatin analog contains both a cell-reactive functional group and GFR. In some aspects, the present invention provides molecular variants of any of the aforementioned cell-reactive analogs of compstatin wherein the cell-reactive functional group or moiety is replaced by a (CH ₂ CH ₂ O) _n moiety (e.g., any of the PEGs described herein) or other a polymer (e.g., POS, polypeptide) with a molecular weight of at least 500 Daltons, e.g., at least 1500 Daltons about 100,000 Daltons (e.g., an average molecular weight of about 20,000; 30,000; 40,000; 50 000; 60,000; 70,000; 80,000; 90,000 or 100,000 Daltons). In some embodiments, the average molecular weight of said compound or moieties (CH ₂ CH ₂ O) _n (or other polymer, eg POS or polypeptide) is at least 20,000 Daltons and up to about 100,000; 120000; 140000; 160000; 180,000 or 200,000 daltons. Thus, it should be understood that the ideas set forth herein regarding cell-reactive compstatin analogs, e.g., useful compstatin analog moieties and linkages by which a compstatin analog moiety is attached to a cell-reactive moiety, can be applied to long-acting compstatin analogs, and the long-acting compstatin analogue can be characterized by any of the structures designated ALM as described above, where A includes a clearance-reducing moiety (e.g., any of the clearance-reducing moieties described herein), and in addition, where one, two, or more may be present. (eg, 3, 4, 5, 6, 7, 8) fragments of the compstatin analogue M, attached to A by connecting parts, referred to herein as L (or L ^P1 , L ^P2 or L ^P3 )). Compstatin analog fragments may contain a peptide whose sequence contains any of SEQ ID NOs: 3-36, 37, 37A, 38A, 39A, 40A, or 41A or variants thereof (e.g., any variant described herein), optionally extended by one or more amino acids at the N-terminus, C-terminus, or both, wherein at least one of the amino acids contains a side chain that contains a reactive functional group such as a primary or secondary amine (e.g., Lys), a sulfhydryl group, a carboxyl group (which may be present as a carboxylate group), a guanidino group, a phenol group, an indole ring, a thioether, or an imidazole ring that facilitates the conjugation of the GFR-containing moiety to the compstatin analog (it is understood that after conjugation, such a reactive functional group will react to form a bond) . It is also understood that where a compstatin analog fragment comprising any of SEQ ID NOs: 3-36, 37, 37A, 38A, 39A, 40A, or 41A or variants thereof is extended by one or more amino acids at the N-terminus, C-terminus, or their both, wherein at least one of the amino acids contains a side chain containing a reactive functional group, such extensions of one or more amino acids can be separated from the cyclic portion of the compstatin analog fragment by a rigid or flexible spacer fragment containing, for example, substituted or unsubstituted, saturated or an unsaturated alkyl chain, an oligo(ethylene glycol) chain, and/or any of the other moieties referred to herein as L (or L ^P1 , L ^P2 or L ^P3 ).

[00281] The following are examples of long-acting analogs of compstatin, characterized in that the value of n is sufficient to provide an average molecular weight of from about 500; 1000; 1500; 2000; 5000; 10,000; 20,000; 30,000; 40,000; 50,000; 60,000; 70,000; 80,000; 90,000 and 100,000 Daltons. In some embodiments, t is sufficient to provide an average molecular weight of from about 20,000 Daltons to about 100,000; 120,000; 140,000; 160,000; 180,000; or 200,000 daltons.

[00282] CH ₂ CH ₂ O) _n C(=O)-Ile-Cys-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys-Thr-NH ₂ (SEQ ID NO:58)

[00283] Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ -C (=O)-CH ₂ CH ₂ O) _n -NH ₂ (SEQ ID NO: 59)

[00284] Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-Lys-C(=O)-CH ₂ CH ₂ O) _n -NH ₂ (SEQ ID NO: 60)

[00285] Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-(Gly) ₅ -Lys-C(=O)-( CH ₂ CH ₂ O) _n -NH ₂ (SEQ ID NO: 61)

[00286] Ac-(CH ₂ CH ₂ O) _n C(=O)-Lys-(Gly) ₅ -Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His- Arg-Cys*-Thr-NH ₂ (SEQ ID NO: 62)

[00287] Ac-(CH ₂ CH ₂ O) _n C(=O)-Lys-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*- Thr-NH ₂ (SEQ ID NO: 63)

[00288] In the sequence of SEQ ID NO: 58, (CH ₂ CH ₂ 0) n is linked by an amide bond to the N-terminal amino acid. In the sequences of SEQ ID NO: 59-63, the specified fragment (CH ₂ CH ₂ O) _n is connected by an amide bond to the side chain of Lys; accordingly, it should be understood that the NH ₂ at the C-terminus in the sequences of SEQ ID NO: 59, 60 and 61 indicates amidation of the C-terminus of the indicated peptide, and in the sequences of SEQ ID NO: 62 and 63 Ac at the N-terminus indicates acetylation N-terminus of said peptide as described above. Those skilled in the art will also appreciate that the free end of the (CH ₂ CH ₂ O) _n moiety typically ends in ( OR ), where the underlined O represents the O atom at the terminal (CH ₂ CH ₂ O) group. Often ( OR ) is a moiety such as hydroxyl ( OH ) or methoxy ( -OCH ₃ ), although other groups (eg, other alkoxy groups) may be used. Accordingly, the sequence of SEQ ID NO: 59, for example, can be represented by the sequence Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH- CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ -C(=O)-Lys-(C(=O)-(CH ₂ CH ₂ O) _n -R)-NH2 (SEQ ID NO: 64) wherein R is is, for example, either H or CH ₃ in the case of a linear PEG. In the case of a bifunctional, branched or star PEG, R is the remainder of said molecule. Further, it should be understood that the fragment containing the reactive functional group may vary as described in this application (for example, corresponding to any of the formulas given in this application).

For example, long-acting compstatin analogues containing the same peptide sequence as SEQ ID NO: 64, in which the reactive functional group-containing moiety contains an ester and/or an alkyl chain, can be represented by the following

Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ -C(=O )-Lys-(C(=O)-(CH2) _m -(CH ₂ CH ₂ O) _n -R-NH2 (SEQ ID NO: 65)

Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ -C(=O )-Lys-(C(=O)-(CH2) _m -(CH ₂ CH ₂ O) _n -R-NH2 (SEQ ID NO: 66)

Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ -C(=O )-Lys-(C(=O)-(CH2) _m -C(=O)-(CH2)j (CH ₂ CH ₂ O) _n -R-NH2 (SEQ ID NO: 67)

In the sequences of SEQ ID NO: 65, 67 m, according to various embodiments, can vary from 1 to about 2, 3, 4, 5, 6, 7, 8, 10, 15, 20, or 30. In the sequences of SEQ ID NO: 67, j according to different implementation options may vary from 1 to about 2, 3, 4, 5, 6, 7, 8, 10, 15, 20, or 30. It should also be borne in mind that as described in this application in various implementations between Lys -(C(=O)- and (CH ₂ CH ₂ O) _n -R other moieties may be introduced such as an amide, an aromatic ring (eg substituted or unsubstituted phenyl) or a substituted or unsubstituted cycloalkyl structure.

[00289] The present invention provides sequence variants of SEQ ID NOs: 58-67 wherein Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr- (SEQ ID NO: 76) is replaced by an amino acid sequence containing the amino acid sequence of any other compstatin analog, such as any of SEQ ID NOs 3-27 or 29-36, 37, 71, 72, 73, 74, or 75, provided that blocking fragment(s) present(s) at the N- and/or C-terminus of the compstatin analog may be absent, replaced by a linker (which may contain a blocking fragment), or attached to a different N- or C-terminal amino acid present in the respective variant(s).

[00290] Any analogue of compstatin, for example, any compound containing any of the sequences of SEQ ID NO: 3-37, 37A, 38A, 39A, 40A or 41A, can according to various implementation options be attached through the N-terminus or C-terminus, or near the N-terminus or C-terminus (e.g., through the amino acid side chain at or near the N-terminal or C-terminal amino acid), directly or indirectly, to any moiety containing a reactive functional group, for example, any compound with formula I XVI or formula AN.

[00291] In some embodiments, the GFR comprises a polypeptide that is present in human serum, or a fragment thereof, or a substantially similar variant of said polypeptide or fragment thereof. In some embodiments, said polypeptide, fragment, or variant has a molecular weight of 5 kDa to 150 kDa, such as at least 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 kDa, or more, for example, from 100 to 120, or from 120 to 150 kDa. In some embodiments, the preparation of a long acting compstatin analog comprises reacting a compstatin analog containing a reactive functional group with one or more amino acid side chains of said polypeptide, wherein said side chain contains a compatible functional group. In some embodiments, the preparation of a long acting compstatin analog comprises reacting a compstatin analog containing a reactive functional group with an N-terminal amine and/or a C-terminal carboxyl group of said polypeptide. In some embodiments, the preparation of a long-acting compstatin analog comprises reacting an amino-reactive functional group-containing compstatin analog with primary amine side chain amino acids (e.g., lysine) and/or the N-terminal amine of said polypeptide. In some embodiments, the preparation of a long acting compstatin analog comprises reacting a compstatin analog containing a carboxyl-reactive functional group with the C-terminal carboxyl group of said polypeptide. In some embodiments, a compstatin analog fragment is attached at each end of said polypeptide and optionally to the side chain of one or more internal amino acids. In some embodiments, the preparation of a long acting compstatin analog comprises reacting a compstatin analog containing a sulfhydryl-reactive functional group with one or more sulfhydryl groups of said polypeptide.

[00292] In some embodiments, at least one reactive functional group is introduced into said polypeptide. For example, in some embodiments, at least one side chain of said polypeptide is modified to convert a first reactive functional group to another reactive functional group prior to reaction with said compstatin analog. In some embodiments, a thiol is administered. Several methods are available for introducing thiols into biomolecules, including the reduction of internal disulfides, as well as the conversion of amines, aldehydes, or carboxylic acid groups to thiol groups. Cystine disulfide crosslinks in proteins can be reduced to cysteine residues using dithiothreitol (DTT/DTT), tris-(2-carboxyethyl)phosphine (TCEP/TCEP), or tris-(2-cyanoethyl)phosphine. Amines can be indirectly thiolated by reaction with succinimidyl 3-(2-pyridyldithio)propionate (SPDP) followed by reduction of the 3-(2-pyridyldithio)propionyl conjugate using DTT or TCEP. Amines can be indirectly thiolated by reaction with succinimidylacetylthioacetate, followed by removal of the acetyl group with 50 mM hydroxylamine or hydrazine at near neutral pH. Amines can be directly thiolated by reaction with 2-iminothiolane, which maintains the overall charge of the indicated molecule and introduces a free thiol. Tryptophan residues in thiol-free proteins can be oxidized to mercaptotryptophan residues, which can then be modified with iodoacetamides or maleimides. A polypeptide containing one or more thiols can be reacted with a compstatin analog containing a maleimide group, such as Ac-Ile-Cys*-Val-Trp(1-Me)-Gln-Asp-Trp-Gly-Ala-His- Arg-Cys*-Thr-AEEAc-Lys-(C(=O)-(CH ₂ ) ₅ -Mal)-NH ₂ (SEQ ID NO: 68), for the synthesis of a long acting compstatin analogue.

[00293] In some embodiments, said polypeptide is produced in a recombinant manner. In some embodiments, said polypeptide is at least partially produced recombinantly (e.g., in bacterial or eukaryotic host cells such as fungal, insect, plant, or vertebrate cells) and/or at least partially produced using chemical synthesis. In some embodiments, said polypeptide is purified. For example, in some embodiments, the polypeptide is purified from a host cell lysate or from a culture medium into which it has been secreted by the host cells. In some embodiments, said polypeptide is glycosylated. In some embodiments, said polypeptide is not glycosylated. In some embodiments, said polypeptide is human serum albumin (HSA). In some embodiments, a substantially similar variant of said polypeptide has sufficient similarity to the polypeptide of which it is a variant not to be recognized as foreign by the subject's normal immune system, e.g., a human subject. In some embodiments, changes in the sequences of a substantially similar variant compared to the polypeptide of which it is a variant are selected to avoid generating MHC class I epitopes. Various techniques known in the art can be used to predict the presence of an MHC class I epitope in a sequence. ways.

[00294] In some embodiments, one or more amino acids in a polypeptide or linker or composition can be chosen to be hydrophobic or hydrophilic, or chosen to impart increased hydrophilicity or, in some embodiments, increased hydrophobicity, connection that contains them. As is known in the art, the terms "hydrophilic" and "hydrophobic" are used to denote the degree of affinity of a substance for water. In some aspects, a hydrophilic substance has a high affinity for water, tending to dissolve in water, mix with water, or be wetted by water, while a hydrophobic substance has substantially no affinity for water and tends to repel and not absorb water, and does not soluble in water, not miscible with water, or not wetted by water. Amino acids can be classified based on their hydrophobicity, as is well known in the art. Examples of "hydrophilic amino acids" are arginine, lysine, threonine, alanine, asparagine, glutamine, aspartate, glutamate, serine, and glycine. Examples of "hydrophobic amino acids" are tryptophan, tyrosine, phenylalanine, methionine, leucine, isoleucine and valine. In certain embodiments, an analog of a standard amino acid is used, wherein said analog has increased or decreased hydrophilicity or hydrophobicity compared to the amino acid of which it is an analog.

[00295] The present invention also provides multimers, e.g., concatemers, containing two or more (e.g., 2 to 10) GFR analogs of compstatin, characterized in that the average molecular weight of the resulting molecule (or its GFR components) is from 20,000; 30,000; 40,000; 50,000; 60,000; 70,000; 80,000; 90,000; up to 100 2000 Daltons. In some embodiments, the average molecular weight of the resulting molecule (or its GFR components) is at least 20,000 Daltons, and up to about 100,000; 120,000; 140,000; 160,000; 180,000; or 200,000 daltons. In some embodiments, said GFR-containing compstatin analogs may be linked via any of the linking moieties described above. Compositions and methods for the preparation of long acting compstatin analogs, as well as synthetic intermediates, are aspects of the present invention.

[00296] In some embodiments, the total molecular weight of the long acting compstatin analog, including fragments of the compstatin analog, is not greater than 50 kDa. For example, in the case of LACA containing 40 kDa PEG, in some embodiments, the molecular weight of the remainder of the compound, including the compstatin analog moiety or moieties, may be no more than 10 kDa, such as 1.5 kDa-5.0 kDa or 5.0 kDa-10 kDa. In some embodiments, the total molecular weight of LACA, including compstatin analog moieties, is between 45 kDa and 50 kDa. In some embodiments, the total molecular weight of LACA, including compstatin analog fragments, is 40 kDa to 45 kDa, 15 kDa to 40 kDa, e.g., 15 kDa to 25 kDa, 25 kDa to 35 kDa, 35 kDa to 40 kDa. Thus, whenever the present specification refers to a compstatin analog containing a polymer or GFR that has a particular molecular weight or has a molecular weight within a particular range, in some embodiments, the total molecular weight of the compstatin analog may exceed the molecular weight of the polymer or GFR, for example, 1.5 kDa-5 kDa, or in some embodiments may exceed the molecular weight of the polymer by 5 kDa-10 kDa. It is understood that the molecular weight of a compound, for example a compound containing a polymer, may mean the average molecular weight of the molecules of such a compound in the composition.

[00297] A wide range of methods and assays suitable for the detection of polymers, for example, PEG, POS and/or polypeptides, and/or suitable for measuring the physical and/or structural properties of polymers, for example, PEG, POS and/or polypeptides, is known in the art. of the art and can, if necessary, be used to detect a compstatin analog, for example, a cell-reactive compstatin analog, a long-acting compstatin analog, a targeted compstatin analog, or a fragment of a compstatin analog. For example, methods and assays suitable for determining properties such as aggregation, solubility, size, structure, melting characteristics, purity, presence of degradation products or contaminants, water content, hydrodynamic radius, and the like are available. Such methods include, for example, analytical centrifugation, various types of chromatography such as liquid chromatography (for example, ion exchange HPLC, size exclusion HPLC, reverse phase HPLC), light scattering, capillary electrophoresis, circular dichroism, isothermal calorimetry, differential scanning calorimetry, fluorescence, infrared (IR), nuclear magnetic resonance (NMR), Raman spectroscopy, refractometry, UV/visible spectroscopy, mass spectrometry, immunological methods, and the like. It should be understood that the methods may be combined. In some aspects, a cell-reactive compstatin analog, a long-acting compstatin analog, or a targeted compstatin analog (or a composition comprising a cell-reactive compstatin analog, a long-acting compstatin analog, or a targeted compstatin analog) has one or more of the properties described herein. , as assessed using any of the above methods. In some aspects, methods suitable for detecting and/or quantifying a long acting compstatin analog are described herein.

[00298] VI. Targeted compstatin analogues

[00299] The present invention provides and/or uses targeted compstatin analogs comprising a targeting moiety and a compstatin analog moiety, characterized in that said targeting moiety non-covalently binds to the target molecule. In some aspects, the present invention provides targeted compstatin analogs similar to the cell-reactive compstatin analogs described in Section VI, characterized in that said compounds contain a targeting moiety in addition to or instead of a cell-reactive moiety. Said targeting fragment may include, for example, an antibody, polypeptide, peptide, nucleic acid (eg, aptamer), carbohydrate, small molecule, or supramolecular complex that specifically binds to the target molecule. In some embodiments, the affinity (as measured by the equilibrium dissociation constant, Kd) of the targeting moiety for the target molecule (as measured by the equilibrium dissociation constant, Kd) is 10 ^-3 M or less, such as 10 ^-4 M or less, such as 10 ^{- 5} M or less, eg 10 ^-6 M or less, 10 ^-7 M or less, 10 ^-8 M or less, or 10 ^-9 M or less under the conditions tested, eg physiological conditions.

[00300] In those embodiments of the present invention in which the targeting fragment is an antibody, said antibody can be any immunoglobulin or derivative thereof that retains binding ability, or any protein containing a binding domain that is homologous or substantially homologous to the immunoglobulin binding domain. Such proteins can be obtained from natural sources, or partially or completely obtained synthetically (for example, using recombinant DNA technologies, chemical synthesis, etc.). Said antibody may belong to any species, for example, human, rodent, rabbit, goat, chicken, etc. Said antibody may belong to any class of immunoglobulins, including any class of human immunoglobulins: IgG, IgM, IgA, IgD, and IgE . According to various embodiments of the present invention, said antibody may be an antibody fragment such as Fab', F(ab').sub.2, scFv (single chain variable), or another fragment that retains an antigen binding site, or a recombinantly produced scFv fragment. , including recombinantly produced fragments. See, for example, Allen, T., Nature Reviews Cancer, Vol. 2, 750-765, 2002 and references incorporated therein. Monovalent, bivalent or multivalent antibodies can be used. Said antibody may be a chimeric antibody, such as one in which a rodent-derived variable domain is fused to a human-derived constant domain, thus retaining the specificity of the rodent antibody. In some embodiments, a human antibody or portion thereof is synthesized, for example, in rodents whose genome contains human immunoglobulin genes, using display technology such as phage display and the like. In some embodiments, a humanized antibody is synthesized by grafting one or more complementarity determining regions from a non-human species (eg, mouse) onto a human antibody sequence. Said antibody may be partially or completely humanized. See, for example, Almagro JC, Fransson J. Humanization of antibodies. Front Biosci. 13:1619-33 (2008) for an overview of various methods for producing humanized antibodies that can be used to generate a targeting fragment suitable for the present invention. The antibody may be polyclonal or monoclonal, although monoclonal antibodies are generally preferred for purposes of the present invention. In certain embodiments of the present invention, an F(ab')2 or F(ab') fragment is used, while in other embodiments, antibodies containing an Fc domain are used. Methods are known in the art for producing specific binding antibodies for virtually any desired molecule. Thus, monoclonal or polyclonal antibodies can be purified from natural sources, for example, from the blood or ascitic fluid of an animal in which the indicated antibody is synthesized (for example, after immunization with the indicated molecule or its antigenic fragment), or can be obtained recombinantly, in cell culture. . Methods for producing antibody fragments are known in the art, for example by cleavage, disulfide reduction or synthesis.

[00301] According to various embodiments of the present invention, the targeting fragment can be any molecule that specifically binds to the target molecule by a mechanism other than antigen-antibody interaction. Such a targeting moiety is referred to as a "ligand". For example, according to various embodiments of the present invention, the ligand may be a polypeptide, peptide, nucleic acid (eg, DNA or RNA), carbohydrate, lipid or phospholipid, or a small molecule. In some embodiments, a small molecule is an organic compound, either naturally occurring or artificially produced, that has a relatively low molecular weight and is not a protein, polypeptide, nucleic acid, or lipid, typically with a molecular weight of less than about 1500 g/mol, and, as a rule, containing several carbon-carbon bonds. In general, an aptamer is an oligonucleotide (e.g., RNA or DNA) optionally containing one or more modified nucleosides (e.g., bases or sugars other than the standard 5 bases (A, G, C, T, U) or sugars (ribose and deoxyribose ), which are most commonly found in RNA and DNA), or modified internucleoside bonds (e.g., non-phosphodiester bonds) which, for example, stabilize said molecule, for example, making it more resistant to nuclease degradation) that binds to a particular protein. In some embodiments, oligonucleotide a is up to about 100 nucleosides in length, eg, 12 to 100 nucleosides. Aptamers can be made using an in vitro evolutionary process called SELEX, and methods are known in the art to make aptamers specific for a protein of interest. See, for example, Brody E N, Gold L. J Biotechnol. March 2000; 74(1):5-13. In some embodiments, a peptidic nucleic acid or a locked nucleic acid is used.

[00302] In certain embodiments of the present invention, the targeting fragment comprises a peptide. In some embodiments, a peptide that binds to a target molecule of interest is identified using a display technology such as phage display, ribosome display, yeast display, and the like.

[00303] Small molecules can be used as ligands. Methods for identifying such ligands are known in the art. For example, small molecule ligands can be identified for numerous proteins of interest using in vitro small molecule library screening, including combinatorial libraries, and computer screening, for example, to identify small organic compounds that bind to protein surface depressions (pockets) (Huang, Z., Pharm. & Ther. 86: 201-215, 2000).

[00304] According to certain embodiments of the present invention, targeting fragments are not proteins or molecules that are commonly used as carriers and conjugated to antigens to stimulate antibody synthesis. Examples are carrier proteins or molecules such as bovine serum albumin, fissure hemocyanin, bovine gamma globulin and diphtheria toxin. In certain embodiments of the present invention, said targeting fragment is not an Fc portion of an immunoglobulin molecule. In some embodiments, the targeting moiety is part of a complex containing one or more additional moieties to which it is covalently or non-covalently attached.

[00305] According to various embodiments of the present invention, the target molecule can be any molecule synthesized by a cell (including any forms expressed on the surface of said cell, or modified forms thereof, at least in part resulting from extracellular modification). In some embodiments, the target molecule is an extracellular substance present in or on the tissue. In some embodiments, the target molecule is characteristic of a particular disease or physiological condition, or characteristic of one or more cell or tissue types. The target molecule is often a molecule at least partially present on the cell surface (e.g., a transmembrane or otherwise membrane-attached protein) such that at least a portion of said molecule is available for binding to an extracellular binding agent, such as an antibody. The target molecule may, but need not be, specific to any cell type. For example, a cell type-specific target molecule is often a protein, peptide, mRNA, lipid, or carbohydrate that is present on cells or in cells of a particular type(s) in greater abundance than on cells or in many other cell types. In some instances, the cell type-specific target molecule is present in detectable amounts only in cells or on cells of a particular cell type of interest. At the same time, it should be borne in mind that a suitable cell-type-specific target molecule need not be absolutely specific for the cell type of interest in order to be considered specific for said cell type. In some embodiments, expression levels of a cell type-specific target molecule are at least 3-fold higher in cells of that cell type than in cells of a reference population, which may be, for example, a mixture containing approximately equal numbers of cells from several (e.g., 5 -10 or more) different tissues or organs. In some embodiments, the cell type-specific target molecule is present in amounts at least 4-5 times, 5-10 times, or greater than 10 times the average expression in the reference population. In some embodiments, detection or measurement of a cell type-specific target molecule allows one of skill in the art to distinguish the cell type or types of interest from many, most, or all other cell types. In general, the presence and/or high abundance of most of the target molecules can be determined using one or more standard techniques such as Northern blotting, in situ hybridization, RT-PCR, sequencing, immunological methods such as immunoblotting, immunodetection (e.g., immunohistochemical), or detection by fluorescence after staining with fluorescently labeled antibodies (eg using FACS), oligonucleotide or cDNA microarrays or membrane chips, microarray protein analysis, mass spectroscopy, and the like.

[00306] In some embodiments, the target molecule is a channel, transporter, receptor, or other molecule at least partially exposed on the cell surface. In some embodiments, the target molecule is an anion transporter or water channel (eg, aquaporin protein).

[00307] In some embodiments, the target molecule is a protein at least partially exposed on the surface of red blood cells, such as a glycophorin (e.g., glycophorin A, B, C, or D) or band 3 protein.

[00308] In some embodiments, said target molecule is a protein at least partially exposed on the surface of endothelial cells. In some embodiments, said target molecule is present on the surface of normal, healthy vessels. In some embodiments, said target molecule is present on the surface of activated endothelial cells. In some embodiments, said target molecule is present on the surface of activated endothelial cells but not on the surface of non-activated endothelial cells. In some embodiments, the target molecule is one whose expression or display is induced by a stimulus such as injury or inflammation. In some embodiments, the target molecule will be recognized as "non-self" by the recipient upon receipt of a transplant containing cells expressing the specified target molecule. In some embodiments, said target molecule is a carbohydrate xenoantigen, antibodies to which are typically found in humans. In some embodiments, said carbohydrate comprises a blood group antigen. In some embodiments, said carbohydrate comprises a xenoantigen, e.g., an alpha-Gal epitope (Galalphal-3Galbetal-(3)4GlcNAc-R) (see, e.g., Macher BA and Galili U. The Galalphal, 3Galbetal, 4GlcNAc-R (alpha- Gal) epitope: a carbohydrate of unique evolution and clinical relevance Biochim Biophys Acta 1780(2):75-88 (2008).

[00309] According to some embodiments of the present invention, the compstatin analogue contains both a targeting moiety and GFR.

[00310] In some embodiments, the targeted compstatin analogue contains multiple targeting moieties, which may be the same or different. Different targeting fragments can bind to the same target molecule or to different target molecules. The present invention provides a targeted compstatin analog that is multivalent in the targeting moiety, the compstatin analog, or both.

[00311] In general, the present invention encompasses any method for preparing a compound containing a compstatin analog moiety and a targeting moiety, and the resulting compounds. In some embodiments, a targeted compstatin analog can be prepared using methods generally similar to those described in Section VI, except that the targeting moiety is used in place of, or in addition to, the cell-reactive moiety. In some embodiments, a targeted compstatin analog containing a peptide as a targeting moiety is synthesized as a polypeptide chain containing a compstatin analog moiety and a peptide targeting moiety. Said polypeptide chain optionally contains one or more spacer peptides between the compstatin analog fragment and the targeting fragment.

[00312] In some embodiments, the targeted compstatin analogue has a molar activity of at least about 10%, 20%, or 30%, e.g., 30% to 40%, 30% to 50%, 30% to 60 %, 30% to 70%, 30% to 80%, 30% to 90% or more, of the activity of the corresponding compstatin analogue having the same amino acid sequence (and, where appropriate, one or more blocking fragments), but not containing the targeting fragment. In some embodiments wherein the targeted compstatin analog comprises multiple compstatin analog moieties, said targeted compstatin analog has a molar activity of at least about 10%, 20%, or 30%, e.g., 30% to 40%, from 30% to 50%, 30% to 60%, 30% to 70%, 30% to 80%, 30% to 90% or more, of the total activity of said compstatin analogue fragments. Compositions and methods for preparing targeted compstatin analogs and synthetic intermediates are aspects of the present invention.

[00313] VII. Application

[00314] Cell-reactive compstatin analogs, long-acting or targeted compstatin analogs have a wide range of uses. In this application, without any limitation of the present invention, certain uses of cell-reactive compstatin analogs, long-acting or targeted compstatin analogs and related aspects of the present invention are described. In some embodiments, a cell-reactive, long-acting, or targeted compstatin analog is administered to a subject suffering from complement-mediated damage or at risk for complement-mediated damage to an organ, tissue, or cells. In some embodiments, the cell-reactive analog of compstatin is brought into contact with an organ, tissue, or cells ex vivo and covalently bound to them. Said organ, tissue or cells are administered to the subject and are protected from damage that would otherwise occur under the action of the recipient's complement system.

[00315] Compstatin analogs that do not covalently bind to cells can be used for the purposes described herein. For example, a compstatin analog modified with a fragment that increases the lifetime of the specified compound in the body, and/or a compstatin analog containing a fragment that targets the specified compstatin analog in relation to a cell type or region sensitive to complement activation can be used; the present invention covers such applications. In some embodiments, a long acting compstatin analog is used. In some embodiments, a compstatin analog containing a targeting moiety is used. In some embodiments, a compstatin analog is used that contains both a moiety that increases the lifetime of said compound in the body and a targeting moiety. to targeted compstatin analogs; and (at least in aspects involving administration of the compstatin analog to the subject) embodiments wherein the compstatin analog that does not contain a targeting moiety or a cell-reactive moiety, optionally a long-acting compstatin analog, is used instead of a cellular -reactive analogue of compstatin or along with it.

[00316] Certain uses of interest include: (1) protection of red blood cells (RBCs) from complement mediated damage in individuals with disorders such as paroxysmal nocturnal hemoglobinuria or atypical hemolytic uremic syndrome or other disorders characterized by complement mediated lysis kkk; (2) protection of transplanted organs, tissues and cells from complement-mediated damage; (3) reduction of ischemic/reperfusion (I/R) injury (eg, in individuals suffering from trauma, vascular obstruction, myocardial infarction, or other situations in which I/R injury may occur); and (4) protection of various body structures (eg, retina) or membranes (eg, synovial membrane) that may be exposed to complement components from complement-mediated damage in any of a variety of complement-mediated disorders. The beneficial effects of inhibiting complement activation on the surface of cells or other body structures are not limited to the direct protection of said cells or structures themselves from direct complement-mediated damage (eg, preventing cell lysis). Thus, inhibition of complement activation using the cellular reactive analogue compstatin may reduce anaphylatoxin synthesis and, as a result, neutrophil influx/activation and other pro-inflammatory manifestations, and/or reduce the potentially damaging release of intracellular contents, thus potentially having a beneficial effect on long-term organ systems or the entire body.

[00317] A. Protection of blood cells

[00318] In some embodiments of the present invention, a cell-reactive compstatin analog, a cell-targeted compstatin analog, and/or a non-targeted compstatin analog (e.g., a long-acting but non-targeted compstatin analog) is used to protect blood cells from complement-mediated damage. Said blood cells may be any cellular component of blood, such as erythrocytes (red blood cells, RCC), leukocytes (white blood cells, 6 RCC) and/or platelets. In some embodiments, a cell-targeted analog of compstatin targets a target molecule displayed on the surface of kKK cells, such as glycophorin or band 3 protein. A variety of disorders are associated with complement-mediated damage to blood cells. Such disorders may develop as a result, for example, of a deficiency or defect in one or more cellular or soluble RBCs in an individual, for example, due to (a) mutation(s) of the gene(s) encoding(s) such proteins; (b) mutation(s) in genes necessary for the synthesis or normal functioning of one or more RBCs, and/or (c) the presence of autoantibodies against one or more RBCs. Complement-mediated lysis of kKK can occur as a result of the presence of autoantibodies to antigens of kKK, which can appear due to a variety of reasons (often idiopathic). Individuals with such mutation(s) in the genes coding for RBC and/or antibodies against RBC or against their own cKK have an increased risk of disorders, including complement-mediated damage to the cKK. Individuals who have had one or more characteristic episodes of the disorder have an increased risk of relapse.

[00319] Paroxysmal nocturnal hemoglobinuria (PNH) is a relatively rare disorder including acquired hemolytic anemia characterized by complement-mediated intravascular hemolysis, hemoglobinuria, bone marrow failure, and thrombophilia (a tendency to form blood clots). It affects approximately 16 individuals per million worldwide, occurs in both sexes and can occur at any age, often affecting young adults (Bessler, M. & Hiken, J., Hematology Am Soc Hematol Educ Program, 104-110 (2008 ); Hillman, P. Hematology Am Soc Hematol Educ Program, 116-123 (2008)). PNH is a chronic debilitating disease with recurrent episodes of acute hemolysis leading to severe clinical manifestations and reduced life expectancy. In addition to anemia, many patients experience abdominal pain, dysphagia, erectile dysfunction and pulmonary hypertension, and are at increased risk of renal failure and thromboembolic complications.

[00320] PNH was first described as a separate phenomenon in the first decade of the 19th century, but it was not until the 1950s, after the discovery of an alternative pathway for complement activation, that the cause of hemolysis in PNH was clearly established (Parker CJ. Paroxysmal nocturnal hemoglobinuria: an historical overview. Hematology Am Soc Hematol Educ Program 93-103 (2008)). Normally, CD55 and CD59 are attached to the cell membrane by glycosylphosphatidylinositol (GPI) anchors (glycolipid structures that anchor certain proteins to the plasma membrane). PNH occurs as a consequence of a non-malignant clonal expansion of a hematopoietic stem cell(s) that has acquired a somatic mutation in the PIGA gene encoding a protein involved in the synthesis of GPI anchors (Takeda J, et al. Deficiency of the GPI anchor caused by a somatic mutation of the PIG-A gene in paroxysmal nocturnal hemoglobinuria Cell 73:703-711 (1993)). The descendants of such stem cells are deficient in GPI-anchored proteins, including CD55 and CD59. This defect renders cells susceptible to complement-mediated lysis of cCK. Diagnosis is often made by flow cytometric analysis using antibodies to GPI-anchored proteins. It allows detecting the deficiency of GPI-anchored proteins on the cell surface, determining the degree of deficiency and the proportion of affected cells (Brodsky RA. Advances in the diagnosis and therapy of paroxysmal nocturnal hemoglobinuria. Blood Rev. 22(2):65-74 (2008). Cells cCCs in type III PNH are completely deficient in GPI-related proteins and are highly complement sensitive, whereas cCCs in type II PNH are partially deficient and less sensitive.FLAER is a fluorescently labeled, inactive variant of proaerolysin (a bacterial toxin that binds GPI anchors), and it is increasingly being used in conjunction with flow cytometry to diagnose PNH.The absence of FLAER binding to granulocytes is sufficient to diagnose PNH.In some embodiments, a cell-reactive analogue of compstatin protects cCC in PNH from deposition of C3b.

[00321] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is administered to a subject suffering from an atypical hemolytic syndrome (aHUS). aHUS is a chronic disorder characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure, and is caused by inappropriate complement activation, often due to mutations in genes encoding complement regulatory proteins (Warwicker, P., et al.. Kidney Int 53, 836 -844 (1998) Kavanagh, D. & Goodship, T. Pediatr Nephrol 25, 2431-2442 (2010) Mutations in the complement factor H (CFH) gene are the most common genetic anomaly among patients with aHUS, and 60-70% of these patients die or reach end-stage renal disease within one year of onset (Kavanagh & Goodship, supra). Mutations of factor I, factor B, C3, factor H-related proteins 1-5, and thrombomodulin have also been described. Other causes of aHUS include the presence of autoantibodies against complement regulatory proteins such as CFH.

[00322] In some embodiments, a cell-reactive compstatin analog, long-acting or targeted compstatin analog is administered to a subject found to have a mutation in factor I, factor B, C3, factor H-related proteins 1-5, or thrombomodulin or antibodies against a complement regulatory protein, eg, CFH, or against a convertase (eg, C3Nef) or complement factor, have been found to be present.

[00323] Complement-mediated hemolysis has been observed in a heterogeneous group of other conditions, including autoimmune hemolytic anemias involving antibodies that bind to cCK and leading to complement-mediated hemolysis. For example, such hemolysis can occur in primary chronic cold agglutinin disease, autoimmune hemolytic anemia with warm agglutinin syndrome (tAIGA), and during certain reactions to drugs and other foreign substances (Berentsen, S., et al., Hematology 12, 361- 370 (2007); Rosse, W.F., Hillmen, P. & Schreiber, A.D. Hematology Am Soc Hematol Educ Program, 48-62 (2004)). In some embodiments of the present invention, the cell-reactive analog of compstatin is administered to a subject suffering from chronic cold agglutinin disease or to a subject who is at risk for chronic cold agglutinin disease. In some embodiments of the present invention, a cell-reactive analog of compstatin is administered to a subject suffering from TAAIHA, or to a subject who is at risk for TAAIHA. In another embodiment, a cell-reactive analog of compstatin is used to treat a subject suffering from HELLP syndrome, or a subject who is at risk of developing HELLP syndrome, which is diagnosed by the presence of hemolysis, elevated liver enzymes, and low platelets, and who is in at least some subjects is associated with mutations in the complement regulatory protein(s) (Fakhouri, F., et al., 112: 4542-4545 (2008)).

[00324] In other embodiments, cell-reactive analogs of compstatin are used to protect cCC or other cellular blood components intended for transfusion into a subject. Some examples of such applications are described in more detail below. As already noted, targeted and/or long-acting compstatin analogues can be used in the methods described above for inhibiting complement-mediated hemolysis and/or damage to cCC. In some embodiments, a long-acting compstatin analogue containing a (CH ₂ CH ₂ O) moiety is used to treat PNH or aHUS.

[00325] B. Transplantation

[00326] Transplantation is an increasingly important method of treatment that provides ways to replace organs and tissues damaged by injury, disease, or other conditions. The kidneys, liver, lungs, pancreas, and heart are among the organs that can be successfully transplanted. Tissues often transplanted include bones, cartilage, tendons, corneas, skin, heart valves, and blood vessels. Pancreatic islet or islet cell transplantation is a promising approach for the treatment of diabetes, eg type I diabetes. For the purposes of the present invention, an organ, tissue, or cell (or population of cells) that is to be transplanted, is being transplanted, or has been transplanted, may be referred to as a "graft". For the purposes of the present invention, a blood transfusion is considered a "transplant".

[00327] During transplantation, the graft is exposed to a variety of disruptive influences and stimuli that can contribute to the development of graft dysfunction and, potentially, graft failure. For example, ischemic/reperfusion (I/R) injury is a common and significant cause of morbidity and mortality for many grafts (particularly parenchymal organs) and may be a major factor in determining the likelihood of graft survival. Graft rejection is one of the major risks associated with transplantation between genetically different individuals and can lead to graft failure and the need to remove said graft from the recipient's body.

[00328] According to some embodiments of the present invention, a cell-reactive compstatin analog, a cell-targeted compstatin analog, and/or a long-acting compstatin analog is used to protect a graft from complement-mediated damage. The cell-targeted analogue of compstatin reacts with the transplant cells, attaches covalently to it, and inhibits complement activation. The cell-targeting analog of compstatin binds to a target molecule in the graft (eg, expressed by endothelial cells or other cells in the graft) and inhibits complement activation. The target molecule may, for example, be a molecule whose expression is induced or stimulated by a stimulus such as injury or inflammation, a molecule that is recognized as "non-self" by the recipient, a carbohydrate xenoantigen against which humans normally have antibodies, such as a blood group antigen, or a xenoantigen, for example, a molecule containing an alpha-Gal epitope. In some embodiments, the reduction in complement activation may manifest as a decrease in mean C4d deposition in the blood vessels of grafts that have been contacted with a compstatin analog, e.g., a cell-reactive compstatin analog, compared to the mean C4d deposition in grafts that have not been brought into contact with a compstatin analogue (eg, in subjects with similar transplants receiving other similar therapy).

[00329] The graft may be contacted with a long-acting or targeted cell-reactive analog of compstatin before, during and/or after transplantation, in accordance with various embodiments of the present invention. For example, prior to transplantation, a graft removed from a donor's body may be contacted with a fluid containing a cell-reactive compstatin analogue, a long-acting or targeted compstatin analogue. For example, said graft may be immersed in said solution and/or perfused with said solution. In another embodiment, the cell-reactive compstatin analog, long-acting or targeted compstatin analog is administered to the donor prior to graft retrieval. In some embodiments, the cell-reactive, long-acting or targeted compstatin analog is administered to the recipient during and/or after graft administration. In some embodiments, cell-reactive compstatin, a long-acting or targeted analog, is administered topically to the transplanted graft. In some embodiments, the cell-reactive analogue of compstatin is administered systemically, for example intravenously.

[00330] The present invention provides a composition comprising: (a) an isolated graft; and (b) a cell-reactive compstatin analog, a long-acting or targeted compstatin analog. In some embodiments, said composition further comprises a liquid solution suitable for contacting (e.g., suitable for rinsing, rinsing, immersing, perfusing, maintaining, or storing) a graft (e.g., an organ), in particular an isolated graft removed from the body. donor and intended for transplantation to the recipient. In some embodiments, the present invention provides a composition comprising: (a) a liquid solution suitable for contact with a graft (eg, organ); and (b) a cell-reactive compstatin analogue, a long-acting or targeted compstatin analogue. Said liquid solution may be any liquid solution that is physiologically acceptable to the graft (e.g., an appropriate osmotic formulation, not cytotoxic), medically acceptable in view of subsequent administration of the graft to the recipient (e.g., preferably sterile, or at least free of unacceptable amounts of microorganisms or other contaminants) and compatible with a cell-reactive compstatin analog (i.e., not abolishing the reactivity of said compstatin analog), or compatible with a long-acting or targeted compstatin analog. In some embodiments, the solution is any solution known in the art for any such purpose. In some embodiments, the liquid solution is Marshall's or hyperosmolar citrate solution (Soltran®, Baxter Healthcare), University of Wisconsin-Madison (UW) solution (ViaSpan™, Bristol Myers Squibb), histidine-tryptophan-ketoglutarate (HTK) solution ( Custodial®, Kohler Medical Limited), EuroCollins (Fresenius) and Celsior® (Sangstat Medical), Polysol, IGL-1 or AQIX® RS-1. Of course, other solutions related to physiologically acceptable compositions can be used, for example, containing equivalent or similar ingredients in the same or different concentrations. In some embodiments, the solution does not contain ingredient(s) with which said cell-reactive compstatin analog is expected to actively react, and any solution can be modified or designed to be free of such ingredients. In some embodiments, said cell-reactive compstatin analog is present in the graft-compatible solution at a concentration of, for example, 0.01 mg/mL to 100 mg/mL, or may be added to said solution to provide such a concentration.

[00331] In some embodiments, the present invention provides a kit comprising: (a) a cell-reactive compstatin analog, a long-acting or targeted compstatin analog; and (b) a graft-compatible solution or solid (eg, powdered) components thereof. Said cell-reactive, long-acting or targeted compstatin analogue may be in solid form (eg, powder) or solution, in at least partially dissolved form. In some embodiments, the cell-reactive compstatin analog, the long-acting or targeted compstatin analog, and/or the graft-compatible solution are provided in predetermined amounts such that, when combined, a solution is formed containing said cell-reactive compstatin analog, the compstatin analog. long-acting or targeted action, with a suitable concentration to bring into contact with the graft. In many embodiments, the cell-reactive compstatin analog, the long-acting or targeted compstatin analog, and the graft-compatible solution or solid (eg, powdered) components thereof are in separate containers included in the kit. In some embodiments, both the cell-reactive analog of compstatin and the components of the graft-compatible solution are in solid form (eg, powder), in separate containers, or as a mixture. In some embodiments, said kit contains instructions for use, for example, instructions for adding the cell-reactive compstatin analog, long-acting or targeted compstatin analog to the graft-compatible solution and/or instructions for contacting the graft with the cell-reactive compstatin analog. Said kit does not necessarily contain a label approved by the government agency responsible for the control of products used for transplantation, cell therapy and/or blood transfusion.

[00332] The present invention also provides a method for covalently linking a compstatin analog to an isolated graft, comprising contacting said isolated graft with a cell-reactive compstatin analog. The present invention also provides an isolated graft to which a compstatin analog is covalently attached. Typically, a plurality of compstatin analog molecules are attached to said isolated graft. In some embodiments, the graft is or contains a parenchymal organ, such as a kidney, liver, lung, pancreas, or heart. In some embodiments, the graft is or contains bone, cartilage, fascia, tendon, ligaments, cornea, sclera, pericardium, skin, heart valve, blood vessel, amniotic membrane, or dura mater. In some embodiments, the graft comprises multiple organs, such as a heart-lung or pancreas-kidney transplant. In some embodiments, the graft does not completely include an organ or tissue. In particular, the graft may contain a portion of an organ or tissue, such as a lobe of a liver, a portion of a blood vessel, a skin flap, or a heart valve. In some embodiments, the graft comprises a preparation containing isolated cells or tissue fragments derived from the original tissue but retaining at least some of the tissue architecture, such as pancreatic islets. In some embodiments, the preparation contains isolated cells that are not connected to each other by connective tissue, for example, hematopoietic stem cells or progenitor cells derived from peripheral and / or umbilical cord blood, or whole blood, or any blood product containing cells, for example, erythrocytes ( ccc) or platelets. In some embodiments, the graft is from a deceased donor (eg, after brain death (DBD donor) or after cardiac death). In some embodiments, depending on the particular type of graft, the graft is obtained from a living donor. For example, kidneys, liver sections, blood cells are among the types of transplants that are often obtained from a living donor without undue risk to said donor and in accordance with sound medical practice.

[00333] In some embodiments, the graft is a xenograft (i.e., the donor and recipient are from different species). In some embodiments, the graft is an autograft (ie, a graft transferred from one body part to another body part of the same individual). In some embodiments, the graft is an isograft (i.e., the donor and recipient are genetically identical). In most embodiments, said graft is an allograft (i.e., the donor and recipient are genetically non-identical members of the same species). In the case of an allograft, the donor and recipient may or may not be genetically related (for example, they may be members of the same family). As a rule, the donor and recipient are blood type compatible (at least compatible in the ABO system and optionally in the Rh, Kell and/or other systems of antigenic compatibility of blood cells). The recipient's blood may be screened for allogeneic antibodies against the graft and/or the recipient and the donor, since the presence of such antibodies can lead to hyperacute rejection (i.e., rejection that begins almost immediately, for example, within minutes of the graft starting to come into contact with recipient's blood). The complement dependent cytotoxicity (CDC) assay can be used to screen a subject's serum for anti-HLA antibodies. Serum is incubated with a panel of lymphocytes with a known HLA phenotype. If the serum contains antibodies against HLA molecules on target cells, cell death occurs as a result of complement-mediated lysis. The use of a selected panel of target cells makes it possible to determine the specificity of the detected antibody. Other techniques suitable for determining the presence or absence of anti-HLA antibodies, and optionally determining their HLA specificity, include ELISA assay, flow cytometric assay, microbead technologies (eg, Luminex technology). The methodology for conducting these assays is well known, and there are a variety of commercially available kits for conducting them.

[00334] In some embodiments, a cell-reactive, long-acting, or targeted compstatin analog inhibits complement-mediated rejection. For example, in some embodiments, a cell-reactive, long-acting, or targeted compstatin analog inhibits hyperacute rejection. Hyperacute rejection is due at least in part to antibody-mediated activation of the recipient's complement system via the classical pathway and, as a result, deposition of MAC on the graft. This is usually due to antibodies already present in the recipient that react with the graft. Although it is desirable to try to avoid hyperacute rejection prior to transplantation by ensuring that a compatible graft is selected, this is not always possible, for example due to time and/or resource constraints. In addition, some recipients (e.g., individuals after multiple transfusions, individuals who have previously received transplants, women after multiple pregnancies) may already have such a significant amount of antibodies, including, potentially, antibodies against antigens that are tested against. usually do not carry out that it can be difficult or even almost impossible to select a guaranteed compatible graft in a timely manner. Such individuals have an increased risk of hyperacute rejection.

[00335] In some embodiments, the cell-reactive compstatin analog, long-acting or targeted compstatin analog inhibits acute graft rejection or failure. In this application, "acute rejection" refers to rejection occurring at least 24 hours, usually at least a few days - a week after transplantation, and up to 6 months after transplantation. Acute antibody-mediated rejection (AMR) often involves a dramatic increase in donor-specific alloantibodies (DSA) during the first few weeks after transplantation. Without being bound by a particular theory, it is possible that pre-existing plasma cells and/or the conversion of memory B cells to new plasma cells play a role in increasing DSA synthesis. Such antibodies can cause complement-mediated damage to the graft, which can be inhibited by bringing said graft into contact with a cell-reactive analogue of compstatin. Without being bound by a particular theory, inhibition of complement activation in the graft may reduce leukocyte infiltration (eg, by neutrophils) also contributing to acute graft failure.

[00336] In some embodiments, a cell-reactive, long-acting, or targeted compstatin analog inhibits complement-mediated I/R graft injury. As described in more detail below, I/R damage can occur when tissue is reperfused to which its blood supply has been temporarily interrupted, as occurs in transplanted organs. Reduction of I/R damage reduces the likelihood of acute graft dysfunction or reduces its severity and reduces the likelihood of acute graft failure.

[00337] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog inhibits chronic graft rejection and/or chronic graft failure. In this application, "chronic graft rejection or failure" refers to rejection or failure occurring at least 6 months after transplantation, e.g., 6 months 1, 2, 3, 4, 5 years or more, after transplantation, often after months or years of normal graft functioning. This is due to a chronic inflammatory and immune response against said graft. For the purposes of the present invention, chronic rejection may include chronic allograft vasculopathy, a term used to refer to fibrosis of the internal blood vessels of transplanted tissue. Since the incidence of acute rejection has been reduced by immunosuppressive therapy programs, chronic rejection has become increasingly important as a cause of graft dysfunction and failure. There is increasing evidence that alloantibody synthesis by B cells is an important element in the occurrence of chronic graft rejection and failure (Kwun J. and Knechtle SJ, Transplantation, 88(8):955-61 (2009). Earlier graft damage may represent contributing factor leading to chronic processes, such as fibrosis, which can eventually lead to chronic rejection.Thus, inhibition of such early damage by the use of a cell-reactive analogue of compstatin may delay and/or reduce the onset or severity of chronic transplant rejection.

[00338] In some embodiments, a long-acting compstatin analog is administered to a transplant recipient to inhibit transplant rejection and/or transplant failure.

[00339] C. Ischemic/reperfusion injury

[00340] Ischemic/reperfusion (I/R) injury is an important cause of tissue damage following trauma and other conditions associated with temporary interruption of blood flow, e.g., myocardial infarction, stroke, severe infection, vascular disease, aneurysm repair, cardiac - pulmonary bypass and transplantation.

[00341] In trauma, systemic hypoxemia, hypotension, and local interruption of blood supply resulting from bruising, pressure syndrome, and vascular injury lead to ischemia that damages metabolically active tissues. Restoration of blood supply triggers an intense systemic inflammatory response, often more detrimental than ischemia itself. After reperfusion of the ischemic area, locally synthesized and released factors enter the circulatory system and reach distant areas, sometimes causing significant damage to organs not affected by the initial ischemic stroke, such as the lungs and intestines, leading to single organ dysfunction and multiorgan dysfunction. Complement activation occurs shortly after reperfusion and is a key mediator of post-ischemic injury, acting both directly and indirectly through chemoattractive and stimulatory effects on neutrophils. All three major complement pathways are activated and, acting jointly or independently, are included in I/R-related adverse events affecting many organ systems. In some embodiments of the present invention, a cell-reactive and compstatin analog, long-acting or targeted compstatin analog is administered to a subject who has recently (e.g., within the previous 2, 4, 8, 12, 24, or 48 hours) experienced an injury, e.g., trauma leading to a risk of I/R injury in the specified subject, for example, due to systemic hypoxemia, hypotension and/or local interruption of blood supply. In some embodiments, the cell-reactive analog of compstatin may be administered intravascularly, optionally in a blood vessel that supplies blood to an injured body part, or directly to said body part. In some embodiments, the subject is suffering from spinal cord injury, traumatic brain injury, burns, and/or hemorrhagic shock.

[00342] In some embodiments, a cell-reactive, long-acting, or targeted compstatin analog is administered to a subject before, during, or after a surgical procedure, such as a surgical procedure that is expected to temporarily stop blood flow to a tissue, organ, or part of the body. Examples of such procedures include cardiopulmonary bypass, angioplasty, heart valve repair/replacement, aneurysm repair, or other vascular surgery. Said cell-reactive analogue of compstatin may be administered before, after and/or during a period of time overlapping with the surgical procedure.

[00343] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is administered to a subject with a history of myocardial infarction, thromboembolic stroke, deep vein thrombosis, or pulmonary embolism. Said cell-reactive analog of compstatin may be administered in combination with a thrombolytic agent such as tissue plasminogen activator (tPA) (e.g., alteplase (activase), reteplase (retavase), tenecteplase (TNKase)), anistreplase (eminase), streptokinase (cabikinase, streptase) or urokinase (abbokinase). Said cell-reactive compstatin analog, long-acting or targeted compstatin analog may be administered before, after and/or during a period of time overlapping with the administration of the thrombolytic agent.

[00344] In some embodiments, a cell-reactive, long-acting, or targeted compstatin analog is administered to a subject to treat I/R injury.

[00345] D. Other complement mediated disorders

[00346] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is administered to the eye to treat an ocular disorder such as age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, or uveitis. For example, a cell-reactive compstatin analog, a long-acting compstatin analog, or a targeted compstatin analog can be administered to a vitreous cavity (eg, by intravitreal injection) to treat a subject suffering from or at risk of AMD. In some embodiments, AMD is neovascular (wet) AMD. In some embodiments, AMD is dry AMD. As understood by one of ordinary skill in the art, dry AMD includes geographic atrophy (GA), intermediate AMD, and early AMD. In some embodiments, a subject with HA is treated to slow or halt the progression of the disease. For example; in some embodiments, treating a subject with GA reduces the rate of retinal cell death. Evidence of a reduction in the rate of retinal cell death may be a reduction in the rate of growth of HA lesion in patients treated with LACA compared to controls (eg, patients who receive a sham injection). In some embodiments, the subject is suffering from intermediate AMD. In some embodiments, the subject is suffering from early AMD. In some embodiments, a subject with intermediate or early AMD is treated to slow or halt the progression of the disease. For example, in some embodiments, treating a subject with intermediate AMD may slow or prevent progression to a late form of AMD (neovascular AMD or GA). In some embodiments, treating a subject with early AMD may slow or prevent progression to intermediate AMD. In some embodiments, the eye is characterized by both HA and neovascular AMD. In some embodiments, the eye has HA but does not have wet AMD. In some embodiments, the cell-reactive, long-acting or targeted compstatin analog is administered by intravitreal injection for the treatment of glaucoma, uveitis (eg, posterior uveitis), or diabetic retinopathy. In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is administered to the anterior chamber, for example, to treat anterior uveitis.

[00347] In some embodiments, a cell-reactive, long-acting, or targeted compstatin analogue is used to treat a subject suffering from an autoimmune disease, or a subject who is at risk for an autoimmune disease, e.g., an autoimmune disease mediated by at least partly with antibodies against one or more self-antigens.

[00348] Cell-reactive compstatin analogs, long-acting compstatin analogs, or targeted compstatin analogs can be administered to a synovial cavity, for example, in a subject suffering from arthritis (eg, rheumatoid arthritis). Of course, they can be administered systemically additionally or

[00349] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from an intracerebral hemorrhage or a subject who is at risk of an intracerebral hemorrhage.

[00350] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from myasthenia gravis or a subject who is at risk of developing myasthenia gravis.

[00351] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from neuromyelitis optic nerve (NMO/OND) or a subject who is at risk of developing NON.

[00352] In some embodiments, a cell-reactive, long-acting, or targeted compstatin analog is used to treat a subject suffering from membranous proliferative glomerulonephritis (MPGN), or a subject who is at risk of developing MPGN, e.g., Type I MPGN , type II MPGN or type III MPGN.

[00353] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from a neurodegenerative disease or a subject at risk for a neurodegenerative disease. In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from neuropathic pain, or a subject who is at risk of developing neuropathic pain. In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from rhinosinusitis or nasal polyposis or a subject at risk for rhinosinusitis or nasal polyposis. In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from cancer or a subject at risk of developing cancer. In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from sepsis or a subject at risk of developing sepsis. In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a subject suffering from adult respiratory distress syndrome or a subject at risk of developing adult respiratory distress syndrome.

[00354] In some embodiments, a cell-reactive compstatin analogue, a long-acting or targeted compstatin analogue is used to treat a subject suffering from anaphylaxis or an infusion reaction, or a subject who is at risk for an anaphylaxis or infusion reaction. For example, in some embodiments, the subject may receive pretreatment before, during, or after receiving a drug or vehicle that may cause an anaphylaxis or infusion reaction. In some embodiments, a subject at risk for anaphylaxis, or a subject suffering from an anaphylactic reaction to a food (e.g., peanut, shellfish, or other food allergens), an insect (e.g., bee, wasp) sting, is treated with a cell-reactive a long-acting or targeted-acting compstatin analogue.

[00355] Said cell-reactive compstatin analog, long-acting or targeted compstatin analog may, according to various embodiments of the present invention, be administered topically or systemically.

[00356] In some embodiments, a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is used to treat a respiratory disease such as asthma or chronic obstructive pulmonary disease (COPD) or idiopathic pulmonary fibrosis. Said cell-reactive compstatin analogue, long acting or targeted compstatin analogue may, for example, be administered to the respiratory tract by inhalation, for example, as a dry powder or spray, or may be administered by injection, for example, intravenously, intramuscularly, or subcutaneously, according to various implementation options. In some embodiments, the cell-reactive compstatin analog, long acting or targeted compstatin analog is used to treat severe asthma, eg, asthma that is not adequately controlled by bronchodilators and/or inhaled corticosteroids.

[00357] In some aspects, methods for treating a complement-mediated disorder, such as a chronic complement-mediated disorder, are provided, comprising administering a long-acting complement inhibitor to a subject in need of treatment for said disorder. Said long-acting compstatin analog, in various embodiments, can be any of the long-acting compstatin analogs described herein. In some aspects, methods for treating a Th17-related disorder are provided, comprising administering a long-acting complement inhibitor to a subject in need of treatment for said disorder.

[00358] In some aspects, a "chronic disorder" is a disorder that persists for at least 3 months and/or is considered a chronic disorder in the art. In many embodiments, the chronic disorder persists for at least 6 months, such as at least 1 year or more, such as indefinitely. One of skill in the art will appreciate that at least some of the manifestations of the various chronic disorders may be intermittent and/or may wax and wane in severity over time. A chronic disorder may be progressive, for example, tending to increase in severity or affect large areas over time. This document discusses a number of chronic complement-mediated disorders. A chronic complement-mediated disorder can be any chronic disorder in which complement activation (eg, excessive or inappropriate complement activation) is involved, eg, as a contributing and/or at least partly causative factor. For convenience, disorders are sometimes grouped on the basis of an organ or system, in particular, frequently affected in subjects suffering from said disorder. It should be borne in mind that some disorders may affect several organs or systems, and such classification(s) is in no way limiting. In addition, some manifestations (eg, symptoms) may be observed in subjects suffering from any of a number of different disorders. Non-limiting information regarding disorders of interest in terms of the present invention can be found, for example, in standard therapeutic guidelines such as the Cecil Textbook of Medicine (for example, edition 23), Harrison's Principles of Internal Medicine (for example, edition 17), and/or standard guidelines for specific areas of medicine, specific body systems or organs, and/or specific disorders.

[00359] In some embodiments, the chronic complement-mediated disorder is a Th2-related disorder. As used herein, a Th2-related disorder is a disorder characterized by an excess and/or excess or inappropriate activity of CD4+ helper T cells of the Th2 subtype ("Th2 cells") in an organism or part thereof, e.g., in at least one tissue, organ or structure. For example, there may be a predominance of Th2 cells relative to CD4+ helper T cells of the Th1 subtype ("Th1 cells"), for example, in at least one tissue, organ, or structure affected by the disorder. As is known in the art, Th2 cells typically secrete characteristic cytokines such as interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-13 (IL-13), while Th1 -cells typically secrete interferon-γ (IFN-γ) and tumor necrosis factor β (TNF-β). In some embodiments, a Th2-related disorder is characterized by excessive synthesis and/or amount of IL-4, IL-5, and/or IL-13, e.g., relative to IFN-γ and/or TNF-β, e.g., in at least some of at least one tissue, organ or structure.

[00360] In some embodiments, the chronic complement-mediated disorder is a Th17-related disorder. According to some aspects, described in more detail in the application PCT/US2012/043845, filed June 22, 2012, entitled "Methods of Treating Chronic Disorders with Complement Inhibitors", complement activation and Th17 cells are involved in a cycle that involves dendritic cells and antibodies and contributes to the maintenance of the pathological immunological microenvironment underlying a number of disorders. Without relying on a specific theory, the indicated pathological immunological microenvironment, once having arisen, is self-sustaining and contributes to damage to cells and tissues. In some aspects, long acting compstatin analogs are suitable for the treatment of Th17 related disorders.

[00361] As used herein, a Th17-related disorder is a disorder characterized by an excess and/or excess or inappropriate activity of CD4+ helper T cells of the Th17 subtype ("Th17 cells") in an organism or part thereof, e.g., in at least one tissue, organ or structure. For example, there may be a predominance of Th17 cells relative to Th1 and/or Th2 cells, for example, in at least one tissue, organ, or structure affected by the disorder. In some embodiments, Th17 cell predominance is a relative predominance, for example, the proportion of Th17 cells to Th17 cells and/or the proportion of Th17 cells to Th17 cells is increased relative to normal values. In some embodiments, the proportion of Th17 cells relative to regulatory T cells (regulatory CD4 ⁺ CD25 ⁺ T cells, also referred to as "Treg cells") is increased relative to normal values. Th17 cell formation and/or Th17 cell activation is facilitated by various cytokines, such as interleukin 6 (IL-6), interleukin 21 (IL-21), interleukin 23 (IL-23), and/or interleukin 1β (IL-1β) . The formation of Th17 cells involves the differentiation of progenitor T cells, such as naive CD4+ T cells, into the Th17 phenotype and their maturation into functional Th17 cells. In some embodiments, Th17 cell generation encompasses any aspect of development, proliferation (propagation), survival, and/or maturation of Th17 cells. In some embodiments, a Th17-related disorder is characterized by excessive synthesis and/or amount of IL-6, IL-21, IL-23, and/or IL-1β. Th17 cells typically secrete characteristic cytokines such as interleukin-17A (IL-17A), interleukin-17F (IL-17F), interleukin-21 (IL-21), and interleukin-22 (IL-22). In some embodiments, a Th17-related disorder is characterized by excessive synthesis and/or amount of an effector Th17 cytokine, eg, IL-17A, IL-17F, IL-21, and/or IL-22. In some embodiments, the excess synthesis or amount of the cytokine is detectable in the blood. In some embodiments, the excess synthesis or amount of the cytokine is detectable locally, for example, in at least one tissue, organ, or structure. In some embodiments, the Th17-associated disorder is associated with a reduced amount of Treg and/or a reduced amount of a Treg-associated cytokine. In some embodiments, a Th17 disorder is any chronic inflammatory disease; the term encompasses a range of diseases characterized by a self-sustaining immune attack against various tissues that is presumably no longer associated with the original causative lesion (which may not be known). In some embodiments, the Th17-associated disorder is any autoimmune disease. Most, if not all, "chronic inflammatory diseases" may actually be autoimmune diseases. Examples of Th17-related disorders include inflammatory skin diseases such as psoriasis and atopic dermatitis; systemic scleroderma and sclerosis; inflammatory bowel disease (IBD) (such as Crohn's disease and ulcerative colitis); Behcet's disease; dermatomyositis; polymyositis; multiple sclerosis (PC); dermatitis; meningitis; encephalitis; uveitis; osteoarthritis; lupus nephritis; rheumatoid arthritis (RA), Sjögren's syndrome, multiple sclerosis, vasculitis; inflammatory disorders of the central nervous system (CNS), chronic hepatitis; chronic pancreatitis, glomerulonephritis; sarcoidosis; thyroiditis, abnormal immune responses to tissue/organ transplantation (eg, transplant rejection); COPD, asthma, bronchiolitis, hypersensitivity, pneumonitis, idiopathic pulmonary fibrosis (IPF), periodontitis, and gingivitis. In some embodiments, the Th17 disease is a classic known autoimmune disease such as type I diabetes or psoriasis. In some embodiments, the Th17-related disorder is age-related macular degeneration.

[00362] In some embodiments, a chronic complement-mediated disorder is an IgE-related disorder. As used herein, an "IgE-related disorder" is a disorder characterized by excessive and/or inadequate synthesis and/or quantity of IgE, excessive or inadequate activity of IgE-producing cells (e.g., IgE-producing B cells or plasma cells), and/or excessive and /or inadequate activity of IgE-responding cells such as eosinophils or mast cells. In some embodiments, the IgE-related disorder is characterized by elevated levels of total IgE and/or in some embodiments, allergen-specific IgE in the subject's plasma and/or locally.

[00363] In some embodiments, a chronic complement-mediated disorder is characterized by the presence of autoantibodies and/or immune complexes in the body that can activate complement through, for example, the classical pathway. Autoantibodies can, for example, bind to self antigens, for example, on cells or tissues in the body. In some embodiments, autoantibodies bind to antigens in blood vessels, skin, nerves, muscles, connective tissue, heart, kidney, thyroid, and the like. In some embodiments, the subject suffers from optic neuromyelitis and the subject develops autoantibodies (e.g., IgG autoantibody) against aquaporin 4. In some embodiments, the subject suffers from pemphigoid and the subject develops autoantibodies (e.g., IgG or IgE autoantibody) against structural hemidesmosome component (e.g., transmembrane collagen XVII (BP180 or BPAG2) and/or BP230 plakin family protein (BPAG1). In some embodiments, the subject suffers from a polyneuropathy, e.g., chronic polyneuropathy, such as multifocal motor neuropathy or acute polyneuropathy, and the subject anti-ganglioside autoantibodies are formed that bind to gangliosides present on nerve cell membranes In some embodiments, a chronic complement-mediated disorder is not characterized by the presence of autoantibodies and/or immune complexes.

[00364] In some embodiments, the chronic complement disorder is a respiratory disorder. In some embodiments, the chronic respiratory disorder is asthma or chronic obstructive pulmonary disease (COPD). In some embodiments, the chronic respiratory disorder is pulmonary fibrosis (eg, idiopathic pulmonary fibrosis), radiation-induced lung injury, allergic bronchopulmonary aspergillosis, hypersensitivity pneumonitis (also known as allergic alveolitis), eosinophilic pneumonia, interstitial pneumonia, sarcoid, Wegener's granulomatosis, or obliterating bronchiolitis. In some embodiments, the present invention provides a method of treating a subject in need of treatment of a chronic respiratory disorder, e.g., asthma, COPD, pulmonary fibrosis, radiation-induced lung injury, allergic bronchopulmonary aspergillosis, hypersensitivity pneumonitis (also known as allergic alveolitis), eosinophilic pneumonia , interstitial pneumonia, sarcoid, Wegener's granulomatosis, or bronchiolitis obliterans, said method comprising administering a long acting complement inhibitor to a subject in need of treatment for said disorder.

[00365] In some embodiments, the chronic complement disorder is allergic rhinitis, rhinosinusitis, or nasal polyposis. In some embodiments, the present invention provides a method of treating a subject in need of treatment for allergic rhinitis, rhinosinusitis, or nasal polyposis, said method comprising administering a long acting complement inhibitor to a subject in need of treatment for said disorder.

[00366] In some embodiments, a chronic complement disorder is a disorder affecting the musculoskeletal system. Examples of such disorders include inflammatory conditions of the joints (eg, arthritis such as rheumatoid arthritis or psoriatic arthritis, juvenile chronic arthritis, spondyloarthropathies, Reiter's syndrome, gout). In some embodiments, the musculoskeletal disorder results in symptoms such as pain, stiffness, and/or limited mobility of the affected body part(s). Inflammatory myopathies include dermatomyositis, polymyositis, and various other chronic muscle inflammation disorders of unknown etiology that result in muscle weakness. In some embodiments, the chronic complement disorder is myasthenia gravis. In some embodiments, the present invention provides a method for treating any of the above disorders affecting the musculoskeletal system, said method comprising administering a long acting complement inhibitor to a subject in need of treatment for said disorder.

[00367] In some embodiments, a chronic complement-mediated disorder is a disorder affecting the integumentary system. Examples of such disorders include, for example, atopic dermatitis, psoriasis, pemphigoid, pemphigus, systemic lupus erythematosus, dermatomyositis, scleroderma, sclerodermatomyositis, Sjögren's syndrome, and chronic urticaria. In some aspects, the present invention provides a method for treating any of the above disorders affecting the integumentary system, said method comprising administering a long acting complement inhibitor to a subject in need of treatment for said disorder.

[00368] In some embodiments, a chronic complement disorder affects the nervous system, such as the central nervous system (CNS) and/or the peripheral nervous system (PNS). Examples of such disorders include, for example, multiple sclerosis, other chronic demyelinating diseases (eg, neuromyelitis of the optic nerve), acute or chronic neuropathies caused by autoantibodies against nerve components such as anti-ganglioside antibodies (eg, Guillain-Barré syndrome, multifocal motor neuropathy) , amyotrophic lateral sclerosis, chronic pain, stroke, allergic neuritis, Huntington's disease, Alzheimer's disease and Parkinson's disease. In some embodiments, the present invention provides a method of treating any of the foregoing disorders affecting the nervous system, said method comprising administering a complement inhibitor, e.g. .

[00369] In some embodiments, a chronic complement disorder affects the circulatory system. For example, in some embodiments, said disorder is a vasculitis or other disorder associated with vascular inflammation, eg, blood vessels and/or lymphatic vessels. In some embodiments, the vasculitis is polyarthritis nodosa, Wegener's granulomatosis, giant cell arteritis, Churg-Strauss syndrome, microscopic polyangiitis, Henoch-Schonlein purpura, Takayasu's syndrome, Kawasaki's disease, or Behcet's disease. In some embodiments, the subject, for example, the subject in need of treatment for vasculitis, is positive for anti-neutrophil cytoplasmic antibodies (ANCA).

[00370] In some embodiments, the chronic complement disorder affects the gastrointestinal tract. For example, said disorder may be an inflammatory bowel disease such as Crohn's disease or ulcerative colitis. In some embodiments, the present invention provides a method for treating a chronic complement-mediated disorder affecting the gastrointestinal tract, said method comprising administering a long acting complement inhibitor to a subject in need of treatment for said disorder.

[00371] In some embodiments, the chronic complement disorder is thyroiditis (eg, Hashimoto's thyroiditis, Graves' disease, postpartum thyroiditis), myocarditis, hepatitis (eg, hepatitis C), pancreatitis, glomerulonephritis (eg, membranous proliferative glomerulonephritis, or membranous glomerulonephritis) or panniculitis.

[00372] In some embodiments, the chronic complement disorder affects the kidneys. For example, said disorder may be glomerulonephritis (eg, membranous proliferative glomerulonephritis or membranous glomerulonephritis). In some aspects, such disorders are characterized by the deposition in the kidneys of complement fragments that may be present along with antibodies. For example, in some embodiments, the disorder that affects the kidneys is C3 glomerulopathy, which includes dense deposit disease (DDD) and C3 glomerulonephritis (C3G). In patients suffering from C3 glomerulopathy, there is staining of C3 fragments (eg, iC3b, C3c, C3dg) in the glomeruli, which can be detected using antibodies. In such patients, overactivation of the alternative complement pathway may be observed, for example, due to the presence of autoantibodies stabilizing C3 convertase, such as C3Nef, or anti-factor B or anti-C3b antibodies. C3 glomerulopathy includes conditions in which a mutation in a gene encoding a complement component or complement-like protein is causally associated with kidney disease. Examples include C3 mutated sludge disease and C3 glomerulonephritis with mutations in CFH or CFH related (CFHR) genes, eg CFHR1, CFHR3 and/or CFRH5. In some embodiments, the disorder affecting the kidney is IgA nephropathy, which is characterized by mesangial deposition of IgA, C3, and/or C3 fragments, and, to a lesser extent, IgG and IgM. In some embodiments, the subject may experience overactivation of the alternative and/or lectin pathway.

[00373] According to some embodiments, the present invention provides methods for treating a subject suffering from chronic pain, characterized in that said methods comprise administering a long acting complement inhibitor to a subject in need of such administration. In some embodiments, the subject is suffering from neuropathic pain. Neuropathic pain has been defined as pain initiated or caused by a primary injury or dysfunction of the nervous system, in particular pain arising as a direct consequence of an injury or disease affecting the somatosensory system. For example, neuropathic pain may result from lesions that involve somatosensory pathways with damage to small fibers of peripheral nerves and/or to the spinothalamocortical system in the CNS. In some embodiments, neuropathic pain results from an autoimmune disease (eg, multiple sclerosis), a metabolic disease (eg, diabetes), infection (eg, a viral disease such as shingles or HIV), vascular disease (eg, stroke), trauma (eg, injury, surgery) or cancer. For example, neuropathic pain may be pain that persists after healing of an injury or after the disappearance of a stimulus to peripheral nerve endings, or pain that occurs due to nerve damage. Examples of neuropathic pain conditions or those associated with neuropathic pain include painful diabetic neuropathy, postherpetic neuralgia (eg, pain that persists or recurs at the site of an acute herpes zoster lesion 3 or more months after an acute episode), trigeminal neuralgia, cancer-related neuropathic pain associated with chemotherapy neuropathic pain associated with HIV neuropathic pain (eg, in HIV neuropathy), central/post-stroke neuropathic pain, neuropathy associated with back pain, eg, low back pain (eg, in radiculopathy, such as constriction spinal nerve roots, such as compression of the lumbar roots, which can result from a herniated disc), spinal stenosis, pain from peripheral nerve injury, phantom limb pain, polyneuropathy, pain associated with spinal cord injury, myelopathy, and multiple sclerosis. In certain embodiments of the present invention, a complement inhibitor is administered according to the inventive administration schedule for the treatment of neuropathic pain in a subject with one or more of the conditions mentioned above.

[00374] In some embodiments, the chronic complement disorder is a chronic ocular disorder. In some embodiments, said chronic ocular disorder is characterized by macular degeneration, choroidal neovascularization (CNV), retinal neovascularization (RNR), ocular inflammation, or any combination of the above. Macular degeneration, CNV, NVS and/or ocular inflammation may be a defining and/or diagnostic feature of said disorder. Examples of disorders that are characterized by one or more of these features include, but are not limited to, conditions associated with macular degeneration, diabetic retinopathy, retinopathy of prematurity, proliferative vitreoretinopathy, uveitis, keratitis, conjunctivitis, and scleritis. Conditions associated with macular degeneration include, for example, age-related macular degeneration (AMD). In some embodiments, the subject is in need of treatment for wet AMD. In some embodiments, the subject is in need of treatment for dry AMD. In some embodiments, the subject is in need of treatment for geographic atrophy (GA). In some embodiments, the subject is in need of treatment for ocular inflammation. Ocular inflammation can affect a significant number of structures in the eye, such as the conjunctiva (conjunctivitis), cornea (keratitis), episclera, sclera (scleritis), uveal tract, retina, vasculature, and/or optic nerve. Signs of ocular inflammation may include the presence of inflammation-associated cells such as white blood cells (eg, neutrophils, macrophages) in the eye, the presence of endogenous inflammatory mediator(s), one or more symptoms such as pain in the eye , redness, sensitivity to light, blurred vision and "flies" before the eyes, etc. Uveitis is a general term that refers to inflammation in the choroid (uveal tract) of the eye, for example in any structures of the choroid, including the iris, ciliary body, or choroid. Specific types of uveitis include iritis, iridocyclitis, cyclitis, parsplanitis, and choroiditis. In some embodiments, the subject is in need of treatment for geographic atrophy (GA). In some embodiments, a chronic ocular disorder is an ocular disorder characterized by damage to the optic nerve (eg, optic nerve degeneration), such as glaucoma.

[00375] As noted above, in some embodiments, the chronic respiratory disease is asthma. Information regarding risk factors, epidemiology, pathogenesis, diagnosis, modern methods of modern asthma management, etc., can be found, for example, in the guide: "Expert Panel Report 3: Guidelines for the Diagnosis and Management of Asthma". National Heart Lung and Blood Institute. 2007. http://www.nhlbi.nih.gov/guidelines/asthma/asthgdln.pdf. (“NHLBI Guidelines”; www.nhlbi.nih.gov/guidelines/asthma/asthgdln.htm). Global Initiative for Asthma, Global Strategy for Asthma Management and Prevention 2010" "GINA Report") and/or standard therapeutic guidelines such as Cecil Textbook of Medicine (20th edition), Harrison's Principles of Internal Medicine (17th edition), and/or standard guidelines for pulmonology. Asthma is a chronic inflammatory disorder of the airways in which many cells and cellular elements such as mast cells, eosinophils, T lymphocytes, macrophages, neutrophils and epithelial cells play a role. Individuals suffering from asthma experience recurring episodes associated with symptoms such as obstruction, respiratory failure (also called dyspnea or shortness of breath), chest tightness, and cough. These episodes are usually associated with widespread but variable airway obstruction, often reversible, either spontaneously or with treatment. Inflammation also leads to an associated increase in existing bronchial hyperresponsiveness to a variety of stimuli. Airway hyperresponsiveness (excessive bronchoconstriction in response to stimuli) is a typical symptom of asthma. In general, airflow limitation is due to bronchoconstriction and airway edema. In some patients with asthma, reversibility of air restriction may not be complete. For example, airway remodeling can lead to persistent narrowing of the airways. Structural changes may include thickening of the subbasal membrane, subepithelial fibrosis, hypertrophy and hyperplasia of airway smooth muscle, proliferation and dilation of blood vessels, hyperplasia and hypersecretion of mucosal glands.

[00376] Individuals with asthma may experience exacerbations, identifiable as events characterized by a change in the individual's prior status. Severe asthma exacerbations can be defined as events that require urgent action by the individual and his/her treating physician to prevent a serious adverse outcome such as hospitalization or death from asthma. For example, a severe exacerbation of asthma may require the use of systemic corticosteroids (eg, oral corticosteroids) in a subject whose asthma is usually adequately controlled without PKD, or an increase in the established maintenance dose may be required. Moderate exacerbations of asthma can be defined as events that cause concern to the subject, which indicate the need for a change in treatment, but are not severe. These events are clinically identified as being outside of the subject's usual range of day-to-day asthma variation.

[00377] Current medications for use in asthma generally fall into two general classes: long-term control medications ("control medications") such as inhaled corticosteroids (ICS), oral corticosteroids (OCS), long-acting bronchodilators ( BDD), leukotriene modifiers (eg, leukotriene receptor antagonists or leukotriene synthesis inhibitors, anti-IgE antibodies (omalizumab (Xolair®)), chromolin and nedocromil used to achieve and maintain control of persistent asthma, and rapid-acting drugs such as short-acting bronchodilators (BCD); used to treat acute symptoms and exacerbations. For the purposes of the present invention, these treatment options may be referred to as "conventional therapy." Treatment of exacerbations may also include increasing the dose and / or intensity of control medication therapy. For example, to restore control of asthma, PC course FROM. Current guidelines call for daily administration of a control medication or, in many cases, administration of multiple doses of a control medication daily to subjects with persistent asthma (with the exception of Xolair, which is administered once every 2 or 4 weeks).

[00378] A subject is generally considered to have persistent asthma if said subject has symptoms on average more than twice a week and/or the subject typically uses a fast-acting medication (e.g., CCD) more than twice a week to control symptoms. "Asthma severity" can be classified based on the intensity of treatment required to control a subject's asthma after treatment of relevant comorbidities and optimization of inhalation techniques and regimens (see, for example, GINA Report; Taylor, DR, Eur Respir J 2008; 32:545 -554). Treatment intensity descriptions may be based on recommended medications and doses according to a step-by-step treatment algorithm, which can be found in guidelines such as the NHLBI Guidelines 2007, the GINA Report and their predecessors, and/or standard medical guidelines. For example, asthma can be classified as intermittent, mild, moderate or severe according to the data in Table X, where "treatment" refers to treatment sufficient to provide the best level of asthma control in a subject. (It should be understood that the categories of mild, moderate, and severe asthma generally refer to persistent rather than intermittent asthma.) One of skill in the art will appreciate that Table X is exemplary, and that not all of these medications are available in every healthcare system, which may affect the assessment of asthma severity in some settings. It should also be understood that the classification of mild/moderate asthma may be influenced by other methods under development or new methods. However, the principle of asthma being defined as mild if sufficient control can be achieved with largely low-intensity treatment and severe if high-intensity treatment is required remains applicable. The severity of asthma can also or alternatively be classified based on the inherent intensity of the disease when left untreated (see, for example, NHBLI Guidelines 2007). An assessment can be made based on current spirometry and the patient's description of symptoms over the previous 2-4 weeks. Current impairment rates and future risk can be assessed and included in determining asthma severity. In some embodiments, asthma severity is determined according to FIG. 3.4(a), 3.4(b), 3.4(c) of the NHBLI Guidelines, for individuals aged 0-4, 5-11 or ≥12 years old, respectively.

[00379] "Asthma control" refers to the extent to which asthma symptoms are reduced or eliminated with treatment (pharmacological or non-pharmacological). Asthma control can be assessed based on factors such as frequency of symptoms, presence of symptoms at night, objective measures of lung function such as spirometry (eg, %FEV1 _of predicted, _FEV1 variability, need for CCD to control symptoms). Current impairment rates and future risk can be assessed and included in determining the level of asthma control. In some embodiments, asthma control is determined according to FIG. 4.3(a), 4.3(b) or 4.3(c) of the NHBLI Guidelines, for individuals aged 0-4, 5-11, or ≥12 years old, respectively.

[00380] In general, one skilled in the art can select appropriate methods for determining severity and/or degree of asthma control, and any classification system deemed appropriate by those skilled in the art can be used.

[00381] According to some embodiments of the present invention, a subject suffering from persistent asthma is treated with a complement inhibitor using the regimen of the present invention. In some embodiments, said subject suffers from mild or moderate asthma. In some embodiments, said subject is suffering from severe asthma. In some embodiments, the subject is suffering from asthma that is not adequately controlled by conventional therapy. In some embodiments, the subject suffers from asthma requiring, when treated with conventional therapy, the use of ICS for sufficient control. In some embodiments, the subject suffers from asthma that cannot be adequately controlled despite the use of ICS. In some embodiments, the subject suffers from asthma requiring, when treated with conventional therapy, the use of a PKC to provide adequate control. In some embodiments, the subject suffers from asthma that cannot be adequately controlled despite the use of high-intensity conventional therapy, including PKC. In some embodiments of the present invention, the long acting complement inhibitor is administered as a control medication, or allows the subject to avoid or reduce the dose of a conventional control medication.

[00382] In some embodiments, said subject is suffering from allergic asthma, which is applicable to most asthmatic individuals. In some embodiments, a subject suffering from asthma is considered to have allergic asthma if a non-allergenic asthma trigger (eg, cold, exercise) is unknown and/or identified by standard diagnostic evaluation. In some embodiments, a subject suffering from asthma is considered to have allergic asthma if said subject (i) reproducibly develops asthma symptoms (or worsening of asthma symptoms) after exposure to an allergen, or allergen(s), to which said subject is sensitive. subject; (ii) IgE specific for the allergen or allergen(s) to which the subject is susceptible is observed; (iii) a positive skin prick test is observed for the allergen or for the allergen(s) to which the said subject is sensitive; and/or (iv) other characteristic symptom(s) consistent with atopy are observed, such as allergic rhinitis, eczema, or elevated serum total IgE levels. Keep in mind that a specific allergenic trigger may not be identified; however, it may be suspected or suspected if the subject experiences worsening of symptoms, such as under specific conditions.

[00383] Inhalation allergen stimulation is a technique widely used in the evaluation of allergic respiratory diseases. Inhalation of an allergen results in a cross-reaction between allergen-specific IgE bound to IgE receptors, for example, on mast cells and basophils. There is an activation of the secretory pathways, which leads to the release of mediators of bronchoconstriction and vascular permeability. Individuals with allergic asthma may develop various manifestations after allergen stimulation, such as early asthmatic response (EAR), delayed asthmatic response (LAR), airway hyperresponsiveness (AHR), and airway eosinophilia, all of which can be detected and quantified in according to the known state of the art. For example, airway eosinophilia can be detected by elevated levels of eosinophils in sputum and/or bronchoalveolar lavage fluid. EAR, sometimes referred to as the immediate asthmatic response (IAR), is a response to an allergen challenge upon inhalation that can be detected shortly after inhalation, typically within 10 minutes (min.) after inhalation, for example, by a decrease in FEV ₁ . The EAR typically peaks within 30 minutes. and disappears within 23 hours (h) after stimulation. For example, a subject may be considered to have a "positive" EAR if his/her FEV ₁ decreases by at least 15%, such as at least 20%, within a specified time period, relative to baseline FEV ₁ (where ""original" in this context refers to the state prior to stimulation, e.g., a state equivalent to the normal state of the specified subject in the absence of an exacerbation of asthma and the absence of exposure to allergic stimuli to which the specified subject is sensitive). The onset of delayed asthmatic response (LAR) typically occurs 3-8 hours after stimulation; it is characterized by cellular inflammation of the airways, increased bronchovascular permeability, and mucus secretion. As a rule, it is detected by a decrease in FEV ₁ , the magnitude of which may be greater than that associated with EAR, and potentially more clinically significant. For example, a subject may be considered to have a "positive" LAR if his/her FEV ₁ is reduced by at least 15%, eg, at least 20%, relative to baseline FEV ₁ within a relevant time period. Delayed airway response (DAR) may begin at approximately 26 to 32 hours, peak at approximately 32 and 48 hours, and disappear within approximately 56 hours after stimulation (Pelikan, Z. Ann Allergy Asthma Immunol. 2010, 104(5): 394-404).

[00384] In some embodiments, the chronic respiratory disorder is chronic obstructive pulmonary disease (COPD). COPD encompasses a spectrum of air-restricted conditions that are not completely reversible even with therapy and tend to progress. Symptoms of COPD include dyspnea (breathing failure), decreased exercise tolerance, cough, sputum production, wheezing, and chest tightness. Individuals with COPD may experience episodes of acute (eg, developing in less than a week and often in 24 hours or less) worsening of symptoms (called COPD exacerbations), the frequency and duration of which can vary and are associated with significant complications. They may be triggered by events such as a respiratory tract infection, exposure to toxic particles, or may have an unknown etiology. Smoking is the most common risk factor for developing COPD; other inhalation exposures may also contribute to the development and progression of this disease. The role of genetic factors in COPD is an area of active research. A small percentage of COPD patients have an inherited deficiency of alpha-1 antitrypsin, the major circulating serine protease inhibitor, and this deficiency can lead to a rapidly progressive form of the disease.

[00385] The characteristic pathophysiological features of COPD include narrowing and structural changes in the small airways and destruction of the lung parenchyma (particularly around the alveoli), most commonly as a result of chronic inflammation. The chronic air restriction seen in COPD typically involves a combination of these factors, and the relative importance of their contribution to air restriction symptoms varies from individual to individual. The term "emphysema" refers to the expansion of the air cavities (alveoli) located distal to the terminal bronchioles, with the destruction of their walls. It should be noted that the term "emphysema" is often used in the clinic to refer to the medical condition associated with such pathological changes. Some individuals with COPD suffer from chronic bronchitis, which is defined in clinical terms as a cough with sputum production on most days out of 3 months of the year, for 2 consecutive years. Additional information on risk factors, epidemiology, pathogenesis, diagnosis, and current management of patients with COPD can be found, for example, in the Global Strategy for the Diagnosis, Management, and Prevention of Chronic Obstructive Pulmonary Disease manual. and Prevention in Chronic Obstructive Pulmonary Disease, updated 2009), available from the Global Initiative on Chronic Obstructive Pulmonary Disease website, me. (GOLD) (www.goldcopd.org), also referred to in this document as the "GOLD Report"; American Thoracic Society/European Respiratory Society Guidelines (2004), available on the ATS website at: www.thoracic.org/clinical/copd-guidelines/resources/copddoc.pdf, referred to in this ATC/ERS COPD Guidelines; and standard therapeutic guidelines such as Cecil Textbook of Medicine (20th ^edition ), Harrison's Principles of Internal Medicine ( ^17th edition); and/or standard guidelines for pulmonology.

[00386] In some embodiments, the methods disclosed herein provide for inhibition (interference, disruption) of the DC-Th17-B-Ab-C-DC cycle described above. For example, administration of a complement inhibitor can disrupt the cycle in which complement stimulates DC cells to direct differentiation towards the Th17 phenotype. As a result, the number and/or activity of Th17 cells is reduced, which in turn reduces the degree of Th17-mediated stimulation of B cells and the production of polyclonal antibodies. In some embodiments, these effects lead to a "reboot" of the immunological microenvironment with a shift towards a more normal, less pathological state. As described in Example 1 PCT/US2012/043845 (WO/2012/178083) and USSN 20140371133 (briefly summarized in Example 27 below), evidence supporting the possibility of a prolonged inhibitory effect of complement inhibition on Th17-associated cytokine synthesis was obtained in an asthma model on animals.

[00387] In some embodiments, inhibition of the DC-Th17-B-Ab-C-DC cycle has a disease-modifying effect. Without being bound by a particular theory, inhibition of the DC-Th17-B-Ab-C-DC cycle may not simply treat the symptoms of the disorder, but interact with fundamental pathological mechanisms that may contribute to ongoing tissue damage, even in cases where symptoms are present. under sufficient control, and/or which may contribute to disease exacerbations. In some embodiments, inhibition of the DC-Th17-B-Ab-C-DC cycle results in remission of the chronic disorder. In some embodiments, remission refers to a state of absence or substantially absence of disease activity in a subject with a chronic disorder, in the presence of a recurrence of the disease. In some embodiments, remission can be maintained for an extended period of time (e.g., at least 6 months, e.g., 6-12 months, 12-24 months, or more) with no further therapy, or with a reduced dose or extended dosing interval. . In some aspects, complement inhibition can alter the immunological microenvironment of Th17 cell rich tissue and convert it to a regulatory T cell (Treg) rich microenvironment. This can allow the immune system to "reboot" and enter a state of remission. In some embodiments, remission, for example, may be maintained until a trigger event occurs. The trigger event can be, for example, infection (which can result in the production of polyclonal antibodies that react with both the infectious agent and self protein), exposure to specific environmental conditions (for example, high levels of airborne contaminants such as ozone , or a specific substance or constituents of smoke, e.g. cigarette smoke, allergens), etc. Genetic factors may play a role. For example, individuals who have specific alleles of genes encoding complement components may have a higher baseline level of complement activity, a more reactive complement system, and/or a lower baseline activity of endogenous complement regulatory proteins. In some embodiments, the individual has a genotype associated with an increased risk of AMD. For example, said subject may have a polymorphism in a gene encoding a complement protein or complement regulatory protein, eg, CFH, C3, factor B, said polymorphism being associated with an increased risk of AMD.

[00388] In some embodiments, the immunological microenvironment may begin to progress towards a pathological condition over time, for example, in a subject who has not yet developed symptoms of a chronic disorder, or in a subject who has developed said disorder and who has been treated as described in this document. Such a transition may occur stochastically (e.g., be due at least in part to presumably random fluctuations in antibody levels and/or affinity) and/or as a result of an accumulation of "subthreshold" trigger events that are not of sufficient intensity to trigger a symptomatic outbreak of the disorder.

[00389] In some embodiments, it is contemplated that a relatively short course of a long-acting compstatin analog, eg, 1 week to 6 weeks, eg, about 24 weeks, may provide a lasting benefit. In some embodiments, remission is provided over an extended period of time, such as 1-3 months, 3-6 months, 6-12 months, 12-24 months, or more. In some embodiments, the subject may be monitored and/or prophylactically treated until symptoms return. For example, the subject may receive treatment before or during exposure to the trigger event. In some embodiments, the subject may be monitored, such as monitoring for an increase in a biomarker, e.g., a biomarker containing an indicator of Th17 cells or Th17 cell activity, or complement activation, and the subject may be treated for an increase in such biomarker. See, for example, a more detailed description in PCT/US2012/043845.

[00390] VIII. Compositions and introduction

[00391] The present invention provides and/or uses a variety of compositions comprising a cell-reactive compstatin analog, a long-acting or targeted compstatin analog. According to various implementation options, the composition may have any property or combination of properties described in this application, provided that they are not mutually exclusive. The present invention provides embodiments of such compositions and methods for their use, characterized in that said compstatin analogue is any compstatin analogue.

[00392] In some embodiments, the composition comprises a purified cell-reactive compstatin analogue, a long-acting or targeted compstatin analogue. Purification can be carried out using a variety of methods, which a person skilled in the art can choose based on the required degree of purity of the various components present in the specified composition before purification. For example, filtration, high performance liquid chromatography, affinity chromatography, and/or other methods and combinations thereof may be used. In some embodiments, said composition comprises at least 80%, 85%, 90%, 95%, 98%, 99% or more long acting or targeted cell reactive compstatin analogs as a percentage of the total weight of the compstatin analog. In some embodiments, said composition comprises at least 80%, 85%, 90%, 95%, 98%, 99% or more of a long-acting or targeted cell-reactive compstatin analog as a percentage of total compstatin analog on a mole basis. . In some embodiments, the composition consists or essentially consists of a long-acting or targeted cell-reactive analog of compstatin.

[00393] In some embodiments, a compstatin cell-reactive analog composition and a cell-reactive functional group-containing compound are characterized in that the ratio of the cell-reactive compstatin analog to the cell-reactive functional group-containing compound, in terms of moles is at least 10:1, 20:1, 50:1, 100:1, 500:1, 1000:1 or more. In some embodiments, said composition comprises at least 80%, 85%, 90%, 95%, 98%, 99% or more of a cell-reactive compstatin analog as a percentage of the total weight of the compstatin analog. In some embodiments, said composition comprises at least 80%, 85%, 90%, 95%, 98%, 99% or more of cell-reactive compstatin analog as a percentage of total compstatin analog on a mole basis. In some embodiments, the composition contains at least 80%, 85%, 90%, 95%, 98%, 99% or more of a cell-reactive compstatin analog by weight. In some embodiments, the composition contains at least 80%, 85%, 90%, 95%, 98%, 99% or more of a long acting compstatin analog by weight. In some embodiments, the composition contains at least 80%, 85%, 90%, 95%, 98%, 99% or more of a targeted compstatin analog by weight. In some embodiments, the composition contains at least 80%, 85%, 90%, 95%, 98%, 99% or more of a targeted compstatin analog by weight. In some embodiments, the weight is a dry weight.

[00394] In some aspects, the present invention provides a pharmaceutical grade composition comprising a cell-reactive compstatin analog, a long-acting or targeted compstatin analog. Said pharmaceutical grade composition, in various embodiments, may have any of the above purity characteristics. Said pharmaceutical grade composition is sufficiently free of endotoxins, heavy metals, and unidentified and/or uncharacterized substances to be acceptable, without further purification, for use as a pharmaceutical composition suitable for administration to a human subject, or for the preparation of a pharmaceutical composition for administration. the human subject. In some embodiments, said pharmaceutical grade composition is sterile.

[00395] Suitable formulations, for example, substantially pure formulations of a cell-reactive compstatin analog, a long-acting compstatin analog, or a targeted compstatin analog, or other active agent, can be combined with pharmaceutically acceptable carriers or vehicles, and the like. to obtain a suitable pharmaceutical composition. The term “pharmaceutically acceptable carrier or base” refers to a non-toxic carrier or base that does not interfere with the pharmacological activity of the specified compound with which it is formulated. One of skill in the art will appreciate that a carrier or vehicle is "non-toxic/non-toxic" if it is compatible with administration to a subject in an amount suitable to deliver the compound without undue toxicity. Pharmaceutically acceptable carriers or bases that may be used in the compositions of the present invention include, but are not limited to, water, physiological saline, Ringer's solution, sodium or potassium acetate solution, 5% dextrose, and the like. The specified composition may contain other components suitable for the desired composition, for example, as described in this application. Additional active compounds, for example, compounds independently suitable for the treatment of a subject suffering from a complement-mediated disorder, may also be included in these compositions. The present invention provides such pharmaceutical compositions comprising a cell-reactive compstatin analog, a long-acting or targeted compstatin analog, and optionally a second active agent suitable for treating a subject suffering from a complement-mediated disorder.

[00396] According to some embodiments, the present invention provides a pharmaceutically acceptable composition suitable for human administration, the packaging of which contains a label approved by a government agency responsible for the control of pharmaceutical agents, for example, the United States Food and Drug Administration (FDA ). In some embodiments, the present invention provides a pharmaceutical kit or package comprising: (a) a pharmaceutically acceptable cell-reactive, long-acting, or targeted compstatin analog in solid form; (b) a pharmaceutically acceptable carrier or vehicle. In some embodiments, the solid form is a powder form, such as a lyophilized powder. Said kit or package optionally contains instructions for dissolving the cell-reactive compstatin analog, long-acting or targeted compstatin analog in a vehicle. In some embodiments, a pharmaceutical kit or package is provided. In some embodiments, said package or kit contains an amount of the pharmaceutical composition sufficient for at least a single dose, such as 1 to 200 doses, or any number or sub-range of doses in between. In some embodiments, the package or kit comprises (i) a first container containing a sufficient amount of cell-reactive compstatin analog, long-acting compstatin analog, or targeted compstatin analog for one or more doses; (ii) a second container containing a pharmaceutically acceptable carrier to be combined with the contents of the first vessel to obtain a composition suitable for administration to a subject by, for example, subcutaneous (abbreviated as SC) or intravitreal (i/v/t) injection. In some embodiments, the pharmaceutical package or kit contains one or more needles and optionally one or more syringes. In some embodiments, at least one pre-filled syringe is provided (eg, from 1 to 200, or any sub-range number in between). In some embodiments, one or more unit dosage forms or pre-measured aliquots are provided. In some embodiments, one or more pens or cartridge pens are provided. In some embodiments, instructions for administration are provided, which, in some embodiments, include instructions for self-administration, such as by subcutaneous injection.

[00397] The pharmaceutical composition may be administered to a subject by any suitable route of administration, including, but not limited to, intravenously, intramuscularly, subcutaneously, by inhalation, nasally, intrathecally, intracranially, intraarterially, orally, rectally, transdermally, intradermally, subcutaneously, and etc. In some embodiments, a composition comprising a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is administered intravenously. In some embodiments, a composition comprising a cell-reactive compstatin analog, a long-acting or targeted compstatin analog is administered intra-arterially. Said composition may be administered locally, either into the vascular system supplying the organ or tissue, or extravascularly in the vicinity of the organ or tissue. The term "administering" is to be understood to encompass directly administering a compound or composition to a subject, instructing third parties to administer a compound or composition to a subject, administering or offering a compound or composition to a subject (e.g., for self-administration), self-administration, and, as appropriate, other methods of providing a compound or composition to a subject.

[00398] Pharmaceutical compositions suitable for injection (for example, for intravenous administration) or administration through a pump or catheter generally include sterile aqueous solutions (for water-soluble compositions) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. Sterile solutions can be prepared by incorporating said compound in the correct amount in a suitable solvent, optionally with a single ingredient or a combination of ingredients such as buffers, eg acetates, citrates, lactates or phosphates; agents for correcting tonicity such as sodium chloride or dextrose; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid, glutathione or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; and other suitable ingredients and the like, as needed, followed by filter sterilization. Many physiologically acceptable compounds are known to those skilled in the art that can be included in a pharmaceutical composition. Other suitable compounds include, for example, carbohydrates such as glucose, sucrose, lactose; dextrans; amino acids such as glycine; polyols, for example mannitol or sorbitol. These compounds can, for example, serve as fillers and/or stabilizers, for example for powder, and/or if lyophilization is involved in the preparation or storage process. Surfactant(s), e.g., Tween-80, Pluronic-F108/F68, deoxycholic acid, phosphatidylcholine, etc., may be included in the composition, e.g. to improve solubility or obtain a microemulsion for the delivery of hydrophobic drugs. The pH value can be adjusted if necessary with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral formulation may be contained in ampoules, disposable syringes or infusion bags, or in single or multi-dose vials made of glass or plastic. Preferably the injectable solutions are sterile and do not contain unacceptable amounts of endotoxin.

[00399] Among other things, the present disclosure provides a specific understanding that, in certain contexts, subcutaneous and/or intramuscular injection may be particularly desirable for administering the compstatin analog described herein (eg, LACA). For example, in some embodiments, subcutaneous and/or intramuscular injection may be preferred (over, for example, intravenous delivery), particularly for the treatment of chronic disorders. Alternatively, or in addition, subcutaneous and/or intramuscular injection may be particularly suitable when, for example, it is desired and/or expected that patients will self-administer the compstatin analog composition (e.g., patients do not need to be administered by a healthcare professional, or patients do not rely on administration by a healthcare professional, especially where such administration by a healthcare professional may require a trip to an infusion center).

[00400] In certain specific aspects, the present disclosure encompasses the understanding that, in those embodiments in which the drug is administered daily, it may be desirable to maintain a more constant blood level than with a longer dosing interval, such as weekly or once every 2 weeks. A more constant blood level may be particularly desirable according to some embodiments (eg, treatment of a chronic disorder described herein). The present description of the invention takes into account that daily IV infusion on a long-term basis can be quite inconvenient, especially for a non-hospitalized patient.

[00401] The present specification specifically recognizes that, in some embodiments, subcutaneous administration may be even more effective and/or desirable than intramuscular injection, especially when self-administration is envisaged or such administration is desirable.

[00402] In certain embodiments, the composition described herein, and in particular the composition containing LACA, can be administered using a device that delivers a dose of the pharmaceutical composition by injection, in some embodiments in at least partially automatic operation. after activation. Such a device is referred to in the art as a "pen" or "auto-injector", and these terms are used interchangeably herein. Typically, a pen or auto-injector allows the dose of the pharmaceutical composition contained in the cartridge, reservoir, or syringe to be injected through a hypodermic needle or needles administered automatically or manually, or via a high velocity jet. The pen or auto-injector can be designed to deliver a single dose or multiple doses.

[00403] In some specific embodiments, such a pen or auto-injector is used for intramuscular and/or subcutaneous injection. According to the present specification, a pen or other auto-injector may be particularly suitable for embodiments that use subcutaneous injection. Pens are typically devices that contain (or can be injected with) a drug in an insulated cartridge or reservoir and to which a needle can be attached.

[00404] In certain embodiments, as noted herein, the present specification provides an understanding that SC and/or IM injection may be a particularly desirable method of delivering one or more of the LACAs described herein. For example, certain particularly desirable and/or effective administration regimens have been documented in the present specification, including administration of LACA by injection (eg, by subcutaneous injection); in some embodiments, such an injection is achieved using a syringe pen (eg, which may have been pre-injected with an appropriate dose or volume). Pens can be long-term (and reusable) or single use. A long-acting pen typically uses a replaceable cartridge, which is removed when it becomes empty, and a new cartridge is inserted into the pen. A single-use pen is usually supplied with a cartridge or reservoir pre-filled with medication. When the cartridge or reservoir becomes empty, the pen is discarded. The cartridge or reservoir may contain a single dose or multiple doses. To use the pen, a needle is attached to the pen and inserted into the skin. Typically, a button is pressed to administer a dose, although other activation methods may be used in some embodiments. In certain embodiments, the auto-injector may comprise a spring-loaded syringe, although one of ordinary skill in the art will appreciate that many technologies are available for auto-injection. In certain embodiments, pressing a button or otherwise activating the device automatically inserts the needle and delivers the drug. In certain embodiments, an auto-injector can be designed to insert the needle automatically and/or exactly to the desired depth into subcutaneous tissue. The pen or auto-injector may contain devices such as a dial that allows the user to select or adjust the dose or depth of injection.

[00405] In certain embodiments, the composition described herein, and in particular; the composition containing LACA is administered using a device containing a dual-chamber syringe. One chamber contains a dry drug (eg lyophilized). The second chamber contains a suitable pharmaceutically acceptable carrier. To use the device, the drug is first reconstituted by mixing the contents of the chambers. Mixing can be done in various ways. In certain embodiments, pressing the plunger causes the contents of the chambers to mix, for example by transferring the carrier into the chamber containing the lyophilized drug.

[00406] Thus, a variety of drug delivery devices can be provided containing the composition described herein (e.g., containing LACA), e.g., pre-filled syringes, dual-chamber syringes, long-acting and/or single-use pens, and cartridges. suitable for use with syringe pens. Such devices may contain one or more doses (eg, one or more of any of the number of doses described herein).

[00407] In some embodiments, LACA can be administered, for example, subcutaneously, using a drug delivery device (sometimes referred to simply as a "delivery device") that includes a pump for introducing a liquid composition containing LACA into the subject's body. Obviously, a pump can be any device that moves liquids by mechanical action, as opposed to the conventional manually operated syringe, which is characterized in that the individual who injects the drug (for example, a healthcare professional or a subject who injects himself drug) must directly immerse the plunger into the container containing the drug to inject. It is obvious that the pump can be driven electrically or mechanically, for example, as described in this document. In some aspects, a delivery device comprising a pump may provide convenient administration of suitable doses with a dosing schedule on alternate days, three times a week, twice a week, once every three, four, five, or six days, or once a week, for example as described below. Of course, such delivery devices can also be used for administration according to other administration schedules, eg regimens involving more frequent administration, eg regimens involving daily administration.

[00408] In some embodiments, the delivery device is portable. The portable device, also referred to as "ambulatory" device, is reasonably light in weight, is sized appropriately to allow the subject to move freely during use of the device, and does not require connection to a power pole or mains outlet. In some embodiments, the portable delivery device can be attached to a belt or shoulder strap, or carried in a case that can be attached to a belt or shoulder strap, or can be stowed in a clothing pocket.

[00409] One of ordinary skill in the art will appreciate that the pump can operate in any of a variety of ways and can use a variety of power sources, such as disposable or rechargeable batteries, AC power (e.g., through a wall outlet in a building), compressed gas, or energy stored in a compressed spring or in a stretched expandable elastic chamber. A device that contains fluid in an expandable balloon reservoir and the fluid supply is controlled by pressure from the elastic walls of the balloon reservoir can be referred to as an "elastomer infusion pump".

[00410] In some embodiments, the delivery device comprises a pump and a syringe containing the fluid to be administered, detachably attached to the device, and a drive unit that can be controlled electronically by a controller, arranged to allow the syringe plunger to slide, resulting in infusion of liquid directly or through a flexible tube through a piercing element, such as a needle or cannula, which is inserted into the subject's body under the skin. For example, in some embodiments, the pump may include a motor that turns a screw that pushes a plunger on a syringe containing fluid. Depressing the plunger causes fluid to be expelled from the syringe and injected into the subject's body through the attached piercing element. Examples of pumps are described in, for example, US Pat. Nos. 6,447,487; 6,592,551; 6,645,177; 8,187,228; In the publications of applications for the US Patent No. 20020123740, 20030229311, 20060184123, 20070100281, 20090123309, 20150038906. Crono PID (NDC No. 8423.2000.02), Crono S-PID30 and Crono S-S-PID 50 (NDC No. 8423.2000.0.0) (CAN goal) Medical Technology (Rivoli, Italy)), and the T34™ Ambulatory Syringe Pump and T60™ Ambulatory Syringe Pump (CME Medical, Blackpool, UK) are examples of portable syringe infusion pumps that can be used in some embodiments.

[00411] In some embodiments, the pump may be electronically programmable or controlled. In some embodiments, the pump is not electronically programmable or controlled.

[00412] In some embodiments, the pump uses electricity as a power source. In some embodiments, the pump does not use electricity as a power source. Such a pump may, for example, use a compressed spring or compressed gas as a power source.

[00413] In some embodiments, the pump is a constant pressure pump that exerts constant pressure to depress the barrel of a syringe containing fluid to be administered. An example of a constant pressure pump is the Freedom60® infusion set (RMS Medical Products, Chester, NY). In some embodiments, the FreedomEdge® infusion system (RMS Medical Products) can be used, for example with a syringe capable of holding up to 20 ml, or a syringe capable of holding up to 30 ml. Another example of a constant pressure device is the SCIg60 syringe pump (EMED Technologies, El Dorado Hills, CA). In some embodiments, the valve may control the rate of fluid flow. In some embodiments, a tube attached to a syringe can control the rate of fluid flow, for example, as described in U.S. Patent Applications Nos. 20150374911 and/or 20160256625.

[00414] In some embodiments, the fluid to be administered is contained in a pressurized chamber prior to administration. In some embodiments, the liquid is contained in a resilient, expandable portion of a container, such as a bladder or balloon, prior to delivery. The expandable portion of the container may be made from a compatible butyl, silicone, or other medical grade material suitable for storing fluid, or may have an internal lining of said material. A portion of the container expands upon filling with a liquid (eg, a dosage unit of the compound being administered) to apply pressure to the liquid. One of ordinary skill in the art will appreciate that a portion of a container can be filled in a variety of ways. In some embodiments, filling of the expandable portion of the container may be done manually, for example, using a manually operated syringe, or may be done using a filling device. Once the device has been attached to the subject's skin, a piercing element such as a needle or cannula, which may be provided with a spring, may automatically or upon further activation, for example by pressing a button, move out of the body of the device and pierce the skin. The pressure then automatically or after additional activation, for example, as a result of pressing a button, displaces the liquid from the chamber or container into the subject's body through a needle or cannula. Examples of devices are described in U.S. Patent Application Publication Nos. 20130018326 and/or 20150217058.

[00415] In some embodiments, the delivery device is a "wearable delivery device"; the term refers to a delivery device comprising a chamber or other part of a container for storing a liquid that is administered to a subject, said device being capable of delivering the liquid while being directly attached to the subject's skin without the need for a separate pad or external reservoir, and generally allowing the subject to stay mobile during the delivery process. The liquid storage chamber may be contained in the housing. Typically, the wearable delivery device is attached to the subject's skin with an adhesive. The device is attached firmly enough so that the device is self-supporting. The device may be provided with an adhesive layer, for example, on the outer surface of the housing, which is used to fix the device directly to the skin. An adhesive layer may surround the portion of the device from which the piercing member, such as a needle or cannula, protrudes to provide a seal around the punctured skin. In some embodiments, a wearable delivery device is available from Sensile Medical AG (Hagendorf, Switzerland). For example, devices known as Senselnfuse, SensePatch or Senseflex can be used. In some embodiments, a wearable delivery device is available from Enable Injections, Inc. (Cincinnati, Ohio). In some embodiments, a device that includes a resilient, expandable container portion, such as a bladder or balloon, for ejecting fluid is a wearable delivery device. In some embodiments, the device, such as a wearable delivery device, is designed such that the piercing element, such as a needle, is invisible to the user prior to or during use of the device. In some embodiments, the piercing element, such as a needle, may be retracted when fluid delivery is complete or when the device is detached from the skin. Obviously, a piercing element, such as a needle, for use with the delivery device described herein may be any suitable gauge or inside diameter, such as those gauges or inside diameters as described elsewhere herein.

[00416] In some embodiments, the delivery device includes a housing into which a vial, cartridge, or syringe containing a liquid (eg, a liquid that contains LACA) can be embedded. The fluid is injected after the device is activated. In some embodiments, the liquid is transferred to the chamber of the device prior to administration. In some embodiments, the delivery device is a refillable device, for example, it can be refilled or refilled with a new vial, cartridge, or syringe after the content has been administered.

[00417] It will be apparent to those skilled in the art upon reading this disclosure that, as is standard practice under operating conditions, a container containing a specific volume as described herein may contain an additional volume sufficient to ensure that the designated specific volume (e.g., dosage unit) has been removed from the administration container.

[00418] In some embodiments, the delivery device is a disposable device, i. the device is designed for use with the aim of introducing a single (single) dose or for use in a single injection session. For example, the device may be designed to be attached to a subject's skin, activated to deliver a dose, detached, and then recycled or disposed of instead of being used to deliver one or more additional doses.

[00419] In some embodiments, a delivery device may be used that delivers fluid to two or more sites. In some embodiments, the number of sites is between 1 and 5. In some embodiments, the number of sites is greater than 5, such as between 6 and 10. Delivery to two or more sites may be simultaneous or sequential. The device may contain a pair of syringes, each of which is placed so as to connect with one of the sites and be associated with the housing, which houses the device drive system. Examples of devices are described in WO2011154928 and US Patent Application Publication No. 20120143133. In some embodiments, a multi-needle infusion set may be used. In some embodiments, a multi-needle infusion set comprises a flexible tubing that communicates at one end with a chamber (this term is used interchangeably with the term "reservoir") containing fluid (e.g., a syringe), while the other end branches into a plurality of tubes, at the end of each of which is the needle. Neria™ multi-infusion sets (Unomedical A/S, Östed, Denmark) are examples of multi-needle infusion sets.

[00420] In some embodiments, the delivery device may collect device usage data. Such data may include, for example, the date and time the device was used, delivery parameters such as volume administered, duration of administration, whether any problems occurred during administration, and so on. The data may be stored on a computer-readable medium physically associated with the device and/or may be transmitted to a remote location, such as a remote server, where the data may be stored, analyzed, or transmitted further for storage or analysis. A device may contain one or more processors, sensors, software packages, and associated communications that enable communication between the device and other products and systems. Data can be transferred using radio frequency identification (RFID), barcode/2D code scanning, cellular, Bluetooth low energy (BTLE), physical circuitry, or a combination of these. The data can be transferred via a suitable network, such as the Internet. The data can be analyzed and/or stored in the Cloud. In some embodiments, the device comprises an active or passive RFID tag or chip, hereinafter referred to as an "RFID tag". An RFID tag may contain data that identifies a device. The RFID tag may be an active tag or chip that signals use-related information such as device activation and/or completion of a dose. In some embodiments, data received from a particular device may be made available to one or more organizations or individuals, such as healthcare organizations or their employees, or caregivers of the subject. Such organizations or individuals may additionally or alternatively be automatically notified of the occurrence or non-occurrence of predetermined events. For example, if a dose was not delivered on a day that the administration schedule would require such administration, or if the device was used on a day that the administration schedule would not allow administration, one or more healthcare workers or caregivers may be alerted. subject. Upon notification, the organization or individual can take appropriate action, such as contacting the subject. In some embodiments, the monitoring system automatically attempts to contact the subject, such as by phone or text message, if the dose has not been administered as scheduled.

[00421] In some embodiments, the delivery system may include a delivery device and a remote control device. The remote control device may, for example, provide programming to the delivery device and/or it may be used to activate the delivery device to begin delivery of liquid or to initiate termination of delivery of liquid by the delivery device.

[00422] According to certain embodiments, the present disclosure provides for providing a subject (e.g., by mail or via organized delivery or other regular delivery method) with a set of devices described herein, which together provide a supply of an active agent (e.g., LACA), sufficient to last for a predetermined period of time (eg, one week, two weeks, three weeks, four weeks, etc.). In certain embodiments, such a kit is regularly sent to the patient's residence (eg, every week, two weeks, three weeks, four weeks, etc.) with time frames chosen so that the agent does not run out. In certain embodiments, the composition (eg, LACA) may be contained in a container (eg, vial) or in any of the drug delivery devices or packages mentioned herein. In some embodiments, the supply is sufficient to last for 4 to 12 weeks, 12 to 26 weeks, or more.

[00423] In certain embodiments, the composition is lyophilized or stored refrigerated or frozen immediately prior to administration, during which time it is reconstituted (in the case of a lyophilized composition) or thawed (in the case of a frozen composition). The composition may be brought to room temperature prior to administration.

[00424] In certain embodiments, the composition may be administered using a patch system (eg, for subcutaneous or transdermal administration). In certain embodiments, the patch may comprise a reservoir, or may be attached to a reservoir that may contain, for example, up to about 10 ml of solution, allowing 10 ml of solution to be administered to a patient in a single dose. In some embodiments, the reservoir may contain more than 10 ml, such as 10 ml to 20 ml, 20 ml to 30 ml, 30 ml to 50 ml, or more.

[00425] In certain embodiments, a composition comprising a compstatin analog as described herein, such as a cell-reactive compstatin analog, LACA, or targeted compstatin analog, and a pharmaceutically acceptable carrier has a pH in the range of 6.5 to 7.5 , for example, from 6.8 and 7.2, for example, 7.0. Stability studies have shown that lower pH results in increased stability of certain LACAs compared to neutral pH. Accordingly, in some embodiments, a composition comprising LACA and a pharmaceutically acceptable carrier has a pH of 6.0 to 6.5, 5.5 to 6.0, or 5.0 to 5.5. In certain embodiments, even lower pH may be used, for example between 4.5 and 5.0. In certain embodiments, the composition also contains one or more pharmaceutically acceptable buffering agents suitable for maintaining the pH within a selected range (eg, any of the aforementioned ranges). Suitable buffer substances are described herein (eg acetates, citrates, lactates or phosphates). In certain embodiments, the composition additionally or alternatively contains a salt, for example any of the pharmaceutically acceptable salts described herein. In certain specific embodiments, the composition contains a phosphate.

[00426] It should be noted that, for brevity, certain aspects and embodiments described herein, for example, with respect to administration schedules, doses, routes of administration, devices used for administration, and/or methods of treatment, are described in relation to compstatin analogues. long action. However, it should be understood that similar embodiments are contemplated where the compstatin analogue is not LACA. The compstatin analog may, for example, be a cell-reactive compstatin analog or a targeted compstatin analog that does not contain a clearance-reducing moiety.

[00427] Typically, dispersions are prepared by incorporating the active compound into a sterile base that contains a basic dispersion medium and other suitable ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, preparation methods may include vacuum and freeze drying to obtain a powder of the active ingredient plus any additional desired ingredient, for example, from a pre-filtered sterile solution.

[00428] In certain embodiments, oral administration may be used. Compositions for oral administration typically include an inert diluent or edible carrier. For oral therapeutic administration, the active compound may be formulated with excipients and administered in the form of tablets, lozenges or capsules, eg gelatine capsules. Pharmaceutically compatible binding agents and/or adjuvant materials may be included in the composition. Said tablets, pills, capsules, lozenges, etc. may contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel or corn starch; a lubricant such as magnesium stearate or Sterotes; a lubricant such as colloidal silicon dioxide; a sweetener such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate or orange flavor. A liquid composition may also be administered orally. Formulations for oral delivery may contain agents to improve stability in the gastrointestinal tract and/or to enhance absorption.

[00429] When administered by inhalation, the compstatin analog can be administered in the form of an aerosol spray from a pressurized container or dispenser containing a suitable nebulizing agent, for example a gas such as carbon dioxide. A metered dose inhaler or nebulizer may be used. The aerosol may contain liquid particles or a dry aerosol (eg, dry powders, large porous particles, etc.). Suitable aqueous bases useful in various embodiments include water or saline, optionally including alcohol in some embodiments. In certain embodiments, the composition contains glycerol in water, for example, about 2% glycerol in water. In certain embodiments, the composition contains a surfactant suitable for pulmonary administration. Additionally or alternatively, other excipients suitable for pulmonary administration may be used.

[00430] A variety of different devices are available for administration by inhalation (which is referred to interchangeably as "inhalation administration", "respiratory administration", or "pulmonary administration"). Nebulizers are devices that convert drug solutions or suspensions into aerosols suitable for deposition in the lower respiratory tract. Nebulizer types include jet type nebulizers, ultrasonic wave nebulizers, and electron mesh nebulizers. A partial list of available electronic mesh nebulizers includes eFlow (Pan), i-Neb (Respironics), MicroAir (Omron), IHSO Nebulizer (Beurer) and Aeroneb® (Aerogen). Respimat.®Soft Mist.™ inhaler (Boeringer Ingelheim) can be used. A metered dose inhaler (MDI) is a portable aerosol device that uses an aerosol to deliver a therapeutic agent. MDIs contain a pressurized metal container that contains a pharmaceutical agent in suspension or solution, a nebulizer, a surfactant (typically) and a metering valve. Chlorofluorocarbons (CFCs) have been widely used as propellants, but have been predominantly replaced by hydrofluorocarbons (HFCs, also known as hydrofluoroalkanes (HFAs)), such as HFC-134a and HFC-227ea. Other alternatives are carbon monoxide and nitrogen. A dry powder inhaler (DPI) is a breath-activated device that delivers drug in the form of particles contained in a capsule or blister that is punctured prior to use, usually without the use of an atomizer. Examples of DPIs currently used to deliver drugs for asthma and/or COPD include, for example, Diskus, Aerolizer, HandiHaler, Twisthaler, Flexhaler. Such devices can be used to deliver a compstatin analog in various embodiments. Other exemplary DPI devices that can be used in various embodiments include 3M Conix™, TAIFUN® (AKELA Pharma), Acu-Breathe™ (Respirics), and devices that include Taper Dry Prowder Inhalation technology such as the API-5000, in in which the active pharmaceutical substance (APS) is stored on a microstructured carrier tape.

[00431] Inhalation accessory devices (IAD) are generally divided into 2 categories: spacers and holding chambers. Spacers and holding chambers lengthen the mouthpiece of the inhaler and direct drug mist into the mouth, reducing drug loss to air. The use of an MDI spacer can help reduce the amount of drug that is deposited at the back of the throat, improving the targeting and accumulation of the drug delivered by the MDI. Valved holding chambers (VHCs) allow a cloud of fine drug particles to remain in the spacer until the patient inhales it through a one-way valve, delivering a dose of drug into the lungs. Examples include Aerochamber and Optichamber.

[00432] Particulate compositions can be characterized based on various parameters such as fine particle fraction (FPF), delivered dose, average particle density, and mass median aerodynamic diameter (MMAD). Suitable methods are known in the art, some of which are described in U.S. Patent Nos. 6,942,868 and 7,048,908 and U.S. Publications Nos. MMAD), for example, about 5 µm, although smaller or larger particles can be used. In certain embodiments, particles with a mass median aerodynamic diameter of 1 µm to 25 µm, such as 1 µm to 10 µm, are used.

[00433] A dry particle composition containing particles less than about 1 mm in diameter is also referred to herein as a dry powder. A "dry" composition is characterized by a relatively low liquid content, whereby the particles are readily dispersible, for example in a powder inhalation device to form an aerosol or spray. The "powder" consists primarily or predominantly exclusively of fine solid particles which are relatively free-flowing and capable of being easily dispersed in an inhalation device and then inhaled by the subject, preferably such that a significant proportion of the particles can reach the desired part of the respiratory tract. In certain embodiments, large porous particles are used which have geometric mean diameters ranging from 3 to 15 microns and a bulk density of 0.04 to 0.6 g/cm ³ . See, for example, US patent No. 7048908; Edwards, D. et al. Science 276:1868-1871, 1997; and Vanbever, R., et al., Pharmaceutical Res. 16:1735-1742, 1999).

[00434] In some aspects, various dosages, administration schedules, compositions, and methods are described herein useful for treating patients by inhaled administration of a compstatin analog. The compstatin analog can be any of the compstatin analogs described herein. The compstatin analogue may contain any of the peptides described in Section III of this document. For example, a compstatin analog may comprise or consist of any of SEQ ID Nos: 3-36 described herein and/or may contain any of the modifications compared to compstatin described herein. In specific embodiments, the compstatin analog comprises or consists of a peptide whose sequence is shown in Table 1, for example, SEQ ID NOs: 14, 21, 28, 29, 32, 33, 34, or 36. In specific embodiments, the compstatin analog does not contain a reducing clearance of the fragment, cell-reactive fragment, or targeting fragment. In certain embodiments, the compstatin analog has a molecular weight of 1.5 kDa to 2.0 kDa, or 2.0 kDa to 2.5 kDa. In certain embodiments, the compstatin analog contains 11 to 25 amino acids, for example exactly 13, 14, 15, 16, 17, or 18 amino acids.

[00435] In certain embodiments, the total daily dose for administering a compstatin analog by inhalation (herein, the term is used interchangeably with "pulmonary administration" or "respiratory administration") is at least 5 mg/day, for example, at least 10 mg/day, at least 15 mg/day, at least 20 mg/day, at least 25 mg/day, or at least 30 mg/day. In certain embodiments, the daily dose is 5 mg/day - 20 mg/day, such as 10 mg/day or 15 mg/day. In certain embodiments, the daily dose is from 20 mg/day to 60 mg/day. In certain specific embodiments, the dose is 10 mg/day, 15 mg/day, 20 mg/day, 25 mg/day, 30 mg/day, 35 mg/day, 40 mg/day, 45 mg/day, 50 mg/day. day, 55 mg/day or 60 mg/day. As described in the examples, in a phase 1 clinical study of a compstatin analogue administered to healthy subjects by inhalation, possible evidence of pharmacological activity was observed at doses of 30 mg/day or 60 mg/day.

[00436] In certain embodiments, the daily dose is 60 mg/day to 150 mg/day, such as 60, 70, 80, 90, 100, 110, 120, 130, 140, or 150 mg/day. In certain embodiments, the daily dose is 150 mg/day - 350 mg/day, e.g. , 310, 320, 330, 340, or 350 mg/day. In certain embodiments, the daily dose may be administered in a single administration. In certain embodiments, the daily dose may be administered as two or more doses throughout the day (eg, morning and evening).

[00437] For topical application, the compstatin analogue may be provided as a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical application include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax, and water. Alternatively, pharmaceutically acceptable compositions may be provided as a suitable lotion or cream containing the compstatin analogue suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitol monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[00438] Systemic administration can also be done transmucosally or transdermally. For transmucosal or transdermal administration, penetrants suitable for penetrating such a barrier can be used in said formulation. Such penetrants are well known in the art and include, for example, for transmucosal administration, surfactants, bile salts and fusidic acid derivatives. Transmucosal administration can be carried out, for example, using nasal sprays or suppositories. For transdermal administration, the active compounds are typically formulated into ointments, balms, gels or creams well known in the art.

[00439] These compounds can also be prepared in the form of suppositories (eg, standard suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

[00440] In certain embodiments of the present invention, the compstatin analog or other active compound is formulated with carriers that protect said compound from rapid elimination from the body, such as controlled release formulations such as implants and microencapsulated delivery systems. For example, the compstatin analog may be formulated or encapsulated in a microparticle or nanoparticle formulation. Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, ether polymers, polylactic acid, PLGA, and the like can be used. Liposomes or other lipid-based particles may be used as pharmaceutically acceptable carriers. They can be obtained in accordance with methods known to those skilled in the art, for example, as described in US patent No. 4522811 and/or other sources referenced in this application. Depot formulations containing a compstatin analogue may be used. Said compstatin analog is released from the depot over a period of time, eg, so as to provide a therapeutic concentration over a longer period of time than when the compound is administered intravenously. In some aspects, GFR confers depot properties to the compstatin analog. A person skilled in the art will understand that in order to obtain a controlled release formulation, an implant, and the like. it is necessary to choose materials and methods that allow maintaining the activity of the specified compound.

[00441] It should be appreciated that the compstatin analogue and/or additional active agent(s) may be presented as a pharmaceutically acceptable salt. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentane propionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate , hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate , sulfate, tartrate, thiocyanate, tosylate and undecanoate. Also, pharmaceutically acceptable salts may optionally be prepared as alkali or alkaline earth metal salts, such as sodium, potassium or calcium salts, depending on the properties of the active agent.

[00442] It should be understood that the pharmaceutically acceptable carriers, compounds and methods of preparation referred to in this application are given as examples and are non-limiting. See, for example. Remington: The Science and Practice of Pharmacy. 21st Edition. Philadelphia, PA. Lippincott Williams & Wilkins, 2005 for additional description of pharmaceutically acceptable compounds and methods for preparing various types of pharmaceutical compositions.

[00443] The pharmaceutical composition may be administered in an amount effective to achieve the desired beneficial effect. In some embodiments, the effective amount is sufficient to provide one or more of the following beneficial effects: (i) reduction of at least one symptom or sign of a complement-mediated disorder; (ii) improving the quality of life; (iii) reduce hospitalization rates; (iv) mortality reduction. One of skill in the art will appreciate that the particular beneficial effect will depend at least in part on various factors, such as the particular disorder being treated. One of skill in the art is aware of the symptoms and signs that can be observed in subjects with complement-mediated disorders. Examples of symptoms and signs of various complement-mediated disorders are presented in this document. Thus, in some embodiments, for example, if the subject is suffering from PNH or aHUS, the beneficial effect is a reduction in complement-mediated lysis of red blood cells. In some aspects, the beneficial effect is statistically significant and/or therapeutically significant as judged by one of skill in the art.

[00444] According to certain embodiments of the present invention, a pharmaceutical composition comprising a cell-reactive compstatin analog, a long-acting or targeted compstatin analog, is administered parenterally. In some embodiments, said composition is administered intravenously. In some embodiments, said composition is administered by intravenous injection. In some embodiments, said composition is administered as an IV bolus or IV infusion. In some embodiments, said composition is administered as an IV drip infusion. In some embodiments, said composition is administered as an IV bolus followed by an IV infusion or IV drip. In some embodiments, the IV infusion is administered over about 1, 2, 3, 4, 5, 15, 20, 30, 60, or 120 minutes. In some embodiments, the IV drip is administered over a period of more than about 60 minutes, such as over about 1, 2, 3, or more hours.

[00445] In some embodiments, a total amount of compstatin analog is administered from about 0.1 mg/kg/day to about 2000 mg/kg/day, for example, from about 1 mg/kg/day to about 1000 mg/kg/day. day, for example, from about 5 mg/kg/day to about 500 mg/kg/day. In some embodiments, a total amount of the compstatin analog is administered from about 10 mg/kg/day to about 100 mg/kg/day, for example, from about 10 mg/kg/day to about 50 mg/kg/day, for example, from about 10 mg/kg/day to approximately 20 mg/kg/day. In some embodiments, about 0.5 mg/kg/day to about 10 mg/kg/day of the compstatin analogue is administered. In some embodiments, about 1 mg/kg/day to about 5 mg/kg/day of the compstatin analogue is administered. In some embodiments, about 1 mg/kg/day to about 3 mg/kg/day of the compstatin analogue is administered. In some embodiments, about 3 mg/kg/day to about 5 mg/kg/day of the compstatin analogue is administered. In some embodiments, about 5 mg/kg/day to about 7.5 mg/kg/day of the compstatin analogue is administered. In some embodiments, about 7.5 mg/kg/day to about 10 mg/kg/day of the compstatin analogue is administered. It should be borne in mind that the introduction of the desired total daily amount can be carried out by implementing a variety of administration schemes. For example, the desired amount of compstatin analog can be administered in a single administration or in multiple administrations, eg, over a 24 hour period. For example, a subject may receive two or more doses over a 24 hour period, and said doses may be administered over the same time periods or over different time periods.

[00446] In some embodiments, the cell-reactive compstatin analog, long-acting or targeted compstatin analog is administered at intervals greater than 24 hours. For example, in various embodiments, doses may be administered on an average every other day, every 34 days, weekly, biweekly, and the like. In some embodiments, covalently attached compstatin analogs, long-acting or targeted compstatin analogs protect cells, tissues, organs for a period of weeks or months without the need for repeat treatment. For example, subjects may receive maintenance retreatment at intervals of 1-2 weeks, 2-4 weeks, 4-6 weeks, 6-8 weeks, or even less frequently. In some embodiments, at least some of the doses are administered subcutaneously. For example, some embodiments provide for administering about 0.1-5 mg/kg/day, eg, about 0.5-2 mg/kg/day; for example, in a volume of approximately 0.25 ml - 2 ml, for example, in a volume of approximately 1 ml. In some embodiments, the concentration is about 50 mg/ml - about 300 mg/ml, for example, about 50 mg/ml - about 100 mg/ml or about 100 mg/ml - about 200 mg/ml.

[00447] In some embodiments, administration is daily. According to some variants of implementation of the introduction is carried out 1 or 2 times a day. As detailed in the examples below, daily subcutaneous administration of an example long acting compstatin analog easily achieves blood levels well above 5 μM. In some embodiments, intramuscular administration is used to deliver similar amounts of a compound.

[00448] In some embodiments, a long-acting analog of compstatin is administered to a subject using a therapeutically effective amount, such administration results in blood concentrations of said compound greater than at least 1 μM, at least 2 μM, at least at least 2.5 µM, at least 3 µM, at least 4 µM, at least 5 µM, at least 6 µM, at least 7 µM, at least 8 µM, at least 9 µM, at least at least 10 µM, at least 11 µM, at least 12 µM, or at least 13 µM, at least 14 µM, at least 15 µM, at least 16 µM, at least 18 µM, or at least about 20 µM, or at least about 25 µM, or a level within any range from 4 µM to about 15 µM, or about 20 µM, or about 25 µM. In some embodiments, this level is maintained for at least about 24 hours, or at least about 48 hours, or at least about 72 hours, or at least about 96 hours, or at least about 120 hours, or for at least about 144 hours after a single intravenous injection or after daily subcutaneous injections for about 5-7 days. With prolonged absorption, levels can be maintained for a longer period, for example, up to about 10 days, 12 days, 14 days, or more.

[00449] In some embodiments, the subject receives such treatment to maintain an equilibrium level of about 1.0 µM, about 2.0 µM, about 2.5 µM, about 3.0 µM, about 3.5 µM, about 4 .0 μM, approximately 4.5 μM, approximately 5.0 μM, approximately 5.5 μM, approximately 6.0 μM, approximately 6.5 μM, approximately 7.0 μM, approximately 7.5 μM, approximately 8.0 μM, approximately 8.5 μM, approximately 9.0 μM, approximately 9.5 μM, or approximately 10 μM. In some embodiments, the equilibrium level is from about 1.0 µM to about 10.0 µM, for example, from about 2.0 µM to about 5.0 µM, from about 2.5 µM to about 5.0 µM, from about 5.0 µM to about 7.5 µM, or from about 7.5 µM to about 10 µM, or any value in between within any of the above ranges. In some embodiments, the concentration is sufficient to inhibit substantially the lysis of the red blood cells of PNH patients exposed to human serum in vitro, for example using a modified Ham assay with human serum (see, for example, Example 8). In some embodiments, the concentration is sufficient to reduce at least 50%, 60%, 70%, 80%, 90% or more of red blood cell lysis of PNH patients exposed to human serum in vitro, for example, using a modified Ham analysis with human serum (see, for example, example 8). In some embodiments, the Ham assay can be performed on human serum adjusted to about 0.005 mol/L magnesium and lowered to about 6.2 pH to activate complement. Examples 18 and 19 provide data supporting the ability of the compstatin analogs described herein to inhibit cCC lysis from PNH patients.

[00450] In some aspects, compstatin analogs, for example, long-acting compstatin analogs, can protect the red blood cells of PNH patients from surface accumulation of significant amounts of C3 and/or C3 activation products. Thus, PNH-affected red blood cells that are protected from complement-mediated lysis by compstatin analogs, eg, long-acting compstatin analogs, may also be protected from surface accumulation of significant amounts of C3 and/or C3 activation products. As known in the art, eculizumab (Soliris®, Alexion Pharmaceuticals. Inc.) is a humanized anti-C5 monoclonal antibody approved for the treatment of PNH and aHUS in several countries (see, for example, Dmytrijuk A, FDA report: eculizumab ( Soliris) for the treatment of patients with paroxysmal nocturnal hemoglobinuria Oncologist 2008 Sep;13(9):993-1000 doi: 10.1634/theoncologist.2008-0086 Epub 2008 Sep 10; Westra D., A new era in the diagnosis and treatment of atypical haemolytic uraemic syndrome Neth J Med 2012 Apr;70(3):121-9). It has been reported that when PNH-affected cCC are exposed to eculizumab, accumulation of significant amounts of C3 and/or C3 activation products may be observed on the surface of these cells, which may contribute to their excretion and/or extravascular hemolysis (for example, in the spleen), and, accordingly, may at least partially cause persistent hematologic abnormalities, such as persistent anemia, which occurs in some patients with PNH despite treatment with eculizumab. Without being bound by a particular theory, this may be due to eculizumab's inhibition of MAC formation, protecting cells from MAC-mediated lysis but not inhibiting C3 activation or deposition of C3 and/or C3 activation products, and leaving PNH-affected cells vulnerable to surface C3 and deposition of C3 and/or C3 activation products due to the absence of GPI-anchored complement-inhibiting proteins. Without being bound by a particular theory, the ability of the compstatin analogs described herein to inhibit C3 activation and thus inhibit the production of C3 activation products may provide a significant advantage. In some embodiments, a subject who has received or is receiving treatment with eculizumab and who continues to show signs of hemolysis, e.g., clinically significant hemolysis, e.g., leading to anemia and/or requiring a blood transfusion, is treated with a compstatin analog described herein. In some embodiments, the compstatin analog is used at a concentration sufficient to ensure that the level of C3 and/or C3 activation products on affected PNH cCC exposed to said compstatin analog (in vitro (e.g., Ham assay) or in vivo) is within the range observed in normal kKK from healthy subjects. In some embodiments, the level of C3 and/or C3 activation products on affected PNH cCC exposed to said compstatin analog (in vitro (e.g., Ham assay) or in vivo) is no more than about 1.2; 1.5; 2.0; 2.5; 3.0; 3.5; 4.0; 4.5; or 5.0 from the average level or the upper limit of the normal level. In some embodiments, the level of C3 and/or C3 activation products on affected PNH cCK treated with the compstatin analog (in vitro (e.g., in the Ham assay) or in vivo) is lower than the level of C3 and/or C3 activation products on affected PNH cCK treated with Soliris at a concentration providing equivalent protection against complement-mediated lysis. In some embodiments, the level of C3 and/or C3 activation products on affected PNH cCC exposed to a compstatin analog (in vitro or in vivo) does not exceed about 5%, 10%, 15%, 20%, 25%, 30% , 40% or 50% of C3 levels and/or C3 activation products on affected PNH cCC exposed to eculizumab at a concentration providing equivalent protection against complement-mediated lysis. In some embodiments, the level of C3 and/or C3 activation products on affected PNH cCC exposed to said compstatin analog (in vitro or in vivo) is no more than about 1.2; 1.5; 2.0; 2.5; 3.0; 3.5; 4.0; 4.5; or 5.0 from the mean level or upper limit of the normal range. In some embodiments, the PNH-affected cells comprise or consist of PNH-affected type II cells, PNH-affected type III cells, or a combination thereof. In some embodiments, said kKK are at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or more of type III and/or type II cells. In some embodiments, said cells may contain a number of type I cells. In some embodiments, cCCs can be classified as type I, II, or III cells based on the level of a GPI-anchored protein, such as CD59, on the surface, which can be measured using flow cytometry, immunofluorescence, or ELISA, for example, using an antibody ( e.g., a monoclonal antibody) or other binding agent that binds said GPI-anchored protein. In some aspects, inhibition of C3 deposition and/or C3 activation products on cells or surfaces can be used as an indicator of the efficacy of a compstatin analog in other complement-mediated diseases such as aHUS, other complement-mediated hemolytic diseases, or other complement-mediated diseases. For example, in some embodiments, a compstatin analogue inhibits such deposition on endothelial cells in a subject with aHUS. In some embodiments, said level of C3 and/or C3 activation product(s) can be measured using flow cytometry, immunofluorescence, or ELISA, e.g., using an antibody (e.g., a monoclonal antibody) or other binding agent that binds C3 and/or one or more C3 activation products. In some embodiments, the C3 activation product is C3b, C3c, or C3d. In some embodiments, the binding agent binds C3d. In some embodiments, the binding agent binds C3d and at least one other C3 activation product. In some embodiments, cCC from a PNH patient contacted with an in vitro compstatin analog (e.g., in the Ham assay) is protected from activated complement such that the relative abundance (percentage) of type I and II cells and type III or the relative content or percentage of cells III and I; II and I; or Type III and II are approximately the same as those obtained in the control assay using inactivated complement (eg, heat-inactivated complement). In some embodiments, cCC from a PNH patient contacted with an in vitro compstatin analog (e.g., in the Ham assay) is protected from activated complement such that the relative abundance or percentage of type I and II cells and type III cells, or the relative content or percentage of cells III and I; II and I; or Type III and II differ by no more than 5% from the content or percentage obtained in the control assay using inactivated complement (eg heat-inactivated complement) is used. In some embodiments, complement can be inactivated by heat treatment, which can be implemented by heating complement components or serum or plasma containing complement components to 56° C. or more.

[00451] In some embodiments, to determine the level of lysis, an LDH measurement (lactate dehydrogenase, an enzyme present in significant amounts in red blood cells that can serve as a marker of hemolysis), a measurement of one or more hematological parameters, for example, a measurement of hematocrit, can additionally or alternatively be used to determine the level of lysis , hemoglobin level, reticulocyte count. In some embodiments, one or more of these methods can be used to determine the level of lysis of cKC, e.g., cKC susceptible to complement-mediated lysis, e.g., cells from patients with PNH, cells from patients with aHUS, cells from subjects with other complement-mediated hematological disorders, cells exposed to abnormally high levels of activated complement. In some aspects, provided herein is a method comprising contacting one or more cells in vitro or in vivo with a compstatin analog described herein and measuring the effect of said compstatin analog on one or more indicators of complement-mediated cell damage and/or activation or deposition of complement on the cell surface. In some embodiments, said contact of one or more cells for a sufficient period of time at sufficient concentration results in a decrease in the abnormally high value or an increase in the abnormally low value to values that are within the normal range, or to values that are no more than 5% apart, 10%, 15%, 20%, or 25% of the lower or upper limit of the normal range.

[00452] In some aspects, provided herein is a method for selecting or modifying an administration regimen (dosing regimen) or one or more components of an administration regimen for a patient with a complement-mediated hemolytic disease, such as PNH. Said one or more components of the administration regimen may include a dose, a dosing interval, a route of administration (eg, intravenous or subcutaneous), or a combination thereof. The dose may be a loading dose, a maintenance dose, or both. In some embodiments, one or more blood samples may be obtained from the patient and the schedule for administering the compstatin analog, e.g., a long-acting compstatin analog, or component thereof may be selected or modified to achieve the desired level of protection of the patient's bcc against lysis and /or from the accumulation of C3 and/or product(s) of C3 activation in vitro. In some embodiments, one or more doses of a compstatin analog, such as a long acting compstatin analog, may be administered to a patient, and then one or more blood samples may be obtained from the patient and assessed for C3 levels and/or activation product(s). C3 on the surface. In some embodiments, the administration regimen or component thereof, e.g., dose, dosing interval, or route of administration, may be selected or modified to provide the desired level of protection to the patient's cCC from lysis and/or accumulation of C3 and/or activation products. C3 in vitro or in vivo. In some embodiments, the administration schedule or component thereof, e.g., dose, dosing interval, or route of administration, may be selected or modified to provide the desired level of protection to the patient's cCC from extravascular excretion and/or extravascular lysis in vivo. The desired level may, for example, be represented by a level accepted in the art as providing a clinically significant beneficial effect, a level providing a clinically significant beneficial effect in a particular patient, a level within the normal range, a level selected by a medical professional, or any other level selected. . The normal range for a parameter may be known in the art and/or may be a reference range established by a laboratory, e.g., a clinical laboratory, where the specified value of the relevant parameter when measured is at least 95%, 96%, 97%, 98 % or 99% of the general population, or at least 95%, 96%, 97%, 98%, or 99% of healthy individuals (optionally with matching one or more demographic variables such as sex, age, etc.) or biological samples obtained from them (such as blood samples) is within the specified reference range. The reference range can be established using a population of samples representative of the general population or representative of healthy individuals.

[00453] In some embodiments, the GFR-containing long-acting compstatin analogue is designed to provide slower systemic absorption following subcutaneous or intramuscular administration to a subject compared to a non-GFR compstatin analogue. In some embodiments, specific properties of the GFR, such as length, are chosen to provide a desired rate of systemic absorption following subcutaneous or intramuscular administration compared to at least some other GFRs. In some embodiments, the Cmax is reduced compared to a comparable dose of a non-GFR compstatin analog, thus may contribute to maintaining the plasma concentration of said compound within a desired concentration window, e.g., the therapeutic window. In some embodiments, the long acting compstatin analog composition is characterized in that the subcutaneous dose is completely absorbed within about 1, 2, 3, 4, 6, 8, 12, 15, 30, 45, 60, 90, or 120 hours after injections assessed based on visual inspection of the injection site.

[00454] It should be understood that there may be an initial stage of treatment during which treatment is carried out more frequently and/or higher doses are administered. For example, a subject with PNH or aHUS may need to administer multiple doses to protect a substantial portion of the subject's CCC. Thereafter, lower doses may be administered and/or dosing may be less frequent, for example, to protect newly formed kKK and/or to maintain protection of existing kKK. Of course, similar methods can, in appropriate cases, be used to treat any disease. In some embodiments, treatment begins with IV administration and then moves on to an alternative route, such as subcutaneous, intramuscular, transdermal, or intradermal administration for maintenance therapy. Depending on the disease, treatment may be continued at intervals, for example, for months, years or an unlimited period of time. For example, in some embodiments, a subject may be treated using an administration schedule described herein, such as an administration schedule every other day, three times a week, once every three days, twice a week, or weekly, for a period of time of at least at least 6 weeks, at least 13 weeks, at least 26 weeks, at least 52 weeks or more. In some embodiments, the subject may be treated for 1-2 years, 2-5 years, such as 5 to 10 years, or more. In some embodiments, the disorder is a chronic disorder, which may be a lifetime disorder, and the subject is treated for the period of time that he or she continues to suffer from the disorder, such as for the remainder of the subject's life. Appropriate dosages and administration schedules depend at least in part on the potency and half-life of said compstatin analog (or other active agent), and may optionally be adjusted for the particular recipient, e.g., by administering increasing doses until the desired pre-known response, e.g. complement inhibition and/or cell protection. If necessary, the specific dosage level for any particular subject may be at least in part selected based on a variety of factors including the potency of the particular compound used, the particular condition being treated, age, body weight, general health, route of administration, rate of elimination, combination with any drug; and/or the expression or activity of the complement protein as measured in one or more samples obtained from said subject.

[00455] In some aspects, described herein are methods whereby a patient is switched from treatment with a complement inhibitor that does not contain a compstatin analog to treatment with a compstatin analog, such as a long-acting compstatin analog (LACA), a cell-reactive compstatin analog, or a compstatin analog. targeted action. For brevity, such methods are described in relation to LACA, but alternatively they can be used to transfer a patient to treatment with a cell-reactive compstatin analog or a targeted compstatin analog. In certain embodiments, a patient receiving a compstatin analogue, eg, LACA, is treated with another complement inhibitor that does not contain the compstatin analogue at the time of initiation of therapy with the compstatin analogue, eg, LACA. In embodiments in which the patient receiving the compstatin analog is being treated with a different complement inhibitor that does not contain the compstatin analog, at the time of initiation of therapy with the compstatin analog, the patient's complement system (e.g., as assessed using a suitable assay, such as a serum-induced ex vivo hemolysis using the patient's serum) may already be inhibited at the time of initiation of LACA therapy. In certain embodiments, an excellent complement inhibitor is a C5 inhibitor (ie, an agent that inhibits C5 activation and/or activity, typically by binding to C5). In certain embodiments, the C5 inhibitor is an anti-C5 antibody such as eculizumab, an anti-C5 siRNA such as ALN-CC5 (Alnylam Pharmaceuticals), an anti-C5 polypeptide such as Coversin (Volution Immuno Pharmaceuticals, Ltd.), or an anti-C5 siRNA such as C5. In certain embodiments, an excellent complement inhibitor may be a C3 inhibitor. In certain embodiments, treatment with a different complement inhibitor, such as a C5 inhibitor, is stopped and the patient is switched to LACA treatment. It should be understood that switching to LACA treatment is preferably carried out in such a way that the patient does not experience a clinically significant decrease in the inhibition of the complement system during the transition to treatment with LACA as complement inhibitor monotherapy.

[00456] For example, in the case of a patient with complement-mediated hemolytic anemia, such as PNH, the patient does not experience a clinically significant increase in hemolysis, for example, as measured by LDH and/or ex vivo serum hemolytic activity, when treatment with a different complement inhibitor is stopped. In certain embodiments, the patient is monitored and transitioned such that the patient does not experience a measurable increase in LDH levels greater than 10% or a measurable increase in hemolytic activity greater than 10%. In certain embodiments, the transition to LACA treatment as a single complement inhibitor therapy is carried out so as to maintain at least 80% ex vivo inhibition of serum hemolytic activity during the transition period. "Transition period" means the period of time beginning on the day of the first dose of LACA and ending X days after the last dose of the excellent complement inhibitor times 4, where "X" is the interval between doses at which the excellent complement inhibitor was administered to the patient prior to the first doses of LACA, for example, the recommended interval between doses of an excellent complement inhibitor when used at the recommended dose. "Recommended dose" of a medicinal product means the dose indicated in the instructions for use of this medicinal product on the label approved by the US FDA, or, if the drug is not approved by the FDA, the dose indicated in the instructions for use of the drug on the label approved by the authority responsible for drug approval in the specific jurisdiction in which the drug is used. "Recommended Dosing Interval" for a drug means the dosing interval specified for that drug in the instructions for use and/or on the label approved by the US FDA, or, if the drug is not approved by the FDA, the dose interval specified in the instructions for use and / or on the label approved by the authority responsible for the approval of medicinal products in the particular jurisdiction in which the medicinal product is used. It is understood that a drug may have two or more different recommended administration schedules. Such administration regimens may be suitable for different indications, suitable for patients who have different characteristics, or simply for alternative therapeutically effective combinations of doses, intervals between doses and routes of administration. When a drug has two or more recommended dosing schedules, it is understood that the term "recommended dosing interval" means the recommended dosing interval for a particular dose administered by the particular route of administration used.

[00457] In certain embodiments, LACA is first administered to a patient who has received at least one dose of an excellent complement inhibitor for a period of time not more than 1.5 times the recommended interval between doses of an excellent complement inhibitor, for example, for a period no longer than the recommended dosing interval of the excellent complement inhibitor, for example, no more than half the recommended dosing interval of the excellent complement inhibitor. In certain embodiments, the patient has received at least one dose of a different complement inhibitor within the 2 weeks prior to the patient's first dose of LACA. In certain embodiments, the patient received at least one dose of a different complement inhibitor within 1 week prior to the patient's first dose of LACA. In certain embodiments, the patient received a dose of an excellent complement inhibitor less than a week prior to the patient's first dose of LACA. In certain embodiments, the patient received a dose of a different complement inhibitor on the day of the patient's first dose of LACA. In certain embodiments, the patient is treated with an effective amount of the excellent complement inhibitor and an effective amount of LACA in order to maintain therapeutically effective blood levels of both the excellent complement inhibitor and LACA over a period of time. Such a drug co-administration period may last for, for example, 1 week to 2 weeks, 2 weeks to 4 weeks, 4 weeks to 6 weeks, 6 weeks to 8 weeks, 8 weeks to 12 weeks, 12 weeks to 26 weeks. In certain embodiments, the period of co-administration of drugs may be 1, 2, 3, 4, 5, or 6 months. In certain embodiments, the co-medication period is the period of time from the first dose of LACA to the last dose of a different complement inhibitor. An excellent complement inhibitor can be administered at the usual maintenance dose that the patient received prior to starting LACA treatment. In certain embodiments, the excellent complement inhibitor is administered at a dose that decreases over time (decreases) until the excellent complement inhibitor treatment is eventually discontinued. For example, in some embodiments, the dose of the excellent complement inhibitor can be reduced to about 67% of the maintenance dose, then to about 33% of the maintenance dose for the last dose. In certain embodiments, the dose of the excellent complement inhibitor can be reduced to about 75% of the maintenance dose, then to about 50% of the maintenance dose, then to about 25% of the maintenance dose for the last dose. In certain embodiments, the total period during which a patient is treated with both LACA and a different complement inhibitor (i.e., the time from the first dose of LACA to the last dose of the first complement inhibitor) is between 2 weeks and 26 weeks. In certain embodiments, the dose of the excellent complement inhibitor is not reduced. Instead, in some embodiments, for example, once LACA has reached a therapeutically effective blood level, the blood level of the excellent complement inhibitor decreases as it is metabolized or otherwise eliminated from the body. In certain embodiments, ex vivo serum-induced hemolysis induced by the patient's serum is monitored during and/or after the transition. In certain embodiments, if ex vivo serum-induced hemolysis induced by the patient's serum is not reduced by at least 80%, such as at least 90%, as assessed during the transition from excellent complement inhibitor treatment to LACA treatment as the only complement inhibitor, the dose of LACA can be increased. In certain embodiments, a decrease in opsonization of a patient's red blood cells with PNH fragments of C3, such as C3d, acts as an indicator that the patient is receiving a therapeutically effective amount of LACA. In certain embodiments, treatment with both a different complement inhibitor, eg, a C5 inhibitor, and LACA is continued for at least 6 months, eg, 6 to 12 months, 12 to 24 months, or more (eg, indefinitely). In certain embodiments, a different complement inhibitor and/or LACA is administered at a lower dose than would be used as complement inhibitor monotherapy for a patient. It should be noted that similar methods can be used to switch a patient from LACA treatment to treatment with a different complement inhibitor, if necessary, or to switch a patient from treatment with a first LACA to treatment with a different LACA.

[00458] In some aspects, a subject suffering from any of the complement-mediated disorders described herein, who is being treated with a long-acting or targeted compstatin analog or a compstatin cell-reactive analog, is refractory to treatment with one or more alternative therapies ( ie, therapy options that do not include a long-acting or targeted compstatin analog or a cell-reactive compstatin analog) or demonstrate a suboptimal response to such alternative therapy. In some embodiments, the alternative therapy comprises a corticosteroid. In some embodiments, the alternative therapy comprises an anti-B cell antibody such as rituximab. In some embodiments, the alternative therapy comprises a complement inhibitor, which is a C5 inhibitor, a factor D inhibitor, or a C1 inhibitor. In some embodiments, the alternative therapy comprises an antibody that is a C5 inhibitor, a factor D inhibitor, or a C1 inhibitor.

[00459] In certain aspects, certain dosages, administration schedules, compositions, and methods suitable for administering a long acting compstatin analog (LACA) to a subject are described herein. In some aspects, the specific doses, regimens, compositions, and methods described herein have been used in clinical studies involving subcutaneous or intravitreal administration of LACA containing a 40 kDa linear PEG and two compstatin analog moieties (each attached to each end linear PEG), healthy human subjects or patients with PNH, or in studies involving intravitreal administration of the same LACA in AMD patients (see examples). Accordingly, in certain dosage aspects, the administration schedules, compositions, and/or methods described herein can be used to administer a LACA comprising a linear PEG of approximately 40 kDa and two fragments of a compstatin analog to a subject (e.g., a patient who is suffering from complement-mediated disorder). In certain embodiments, two compstatin analog moieties are attached to the ends of a linear PEG (ie, one compstatin analog moiety is attached to each end of the PEG). In certain embodiments, each compstatin analog moiety is C-terminally attached to a linear PEG. In certain embodiments, the LACA may be CA28-2GS-BF or CA28-2TS-BF.

[00460] As described herein, a fragment of a compstatin analog may contain a spacer that separates an amino acid that contains a side chain containing a reactive functional group, for example, a primary or secondary amine, sulfhydryl, or sulfhydryl reactive group (or an amino acid sequence containing such an amino acid ), from the portion of the compstatin analog fragment containing the cyclic portion, or such linking portion may be absent. If present, such a spacer moiety may contain a substituted or unsubstituted, saturated or unsaturated alkyl chain, an oligo(ethylene glycol) chain, and/or any of the other moieties referred to herein as L (or L ^P1 , L ^P2 or L ^P3 ). When two or more compstatin analog moieties containing spacer moieties are present in a compound, the spacer moieties may be the same or different in various embodiments. Doses, administration regimens, compositions and/or methods can be used to administer long acting compstatin analogues containing compstatin analogue fragments that contain any such spacer fragments.

[00461] As also described herein, in various embodiments, compstatin analog fragments can be attached to PEG (or other polymer) by any of a wide variety of different linkages. In certain embodiments, one or more compstatin analog moieties can be attached to the GFR with an unsaturated alkyl moiety, a non-aromatic cyclic ring system moiety, an aromatic moiety, an ether moiety, an amide moiety, an ester moiety, a carbonyl moiety, an imine moiety, a thioether moiety , an amino acid residue and/or any of the fragments indicated above as L (or L ^P1 , L ^P2 or L ^P3 ). For example, as described herein, in specific embodiments, one or more compstatin analog moieties may be attached to the GFR by a carbamate bond, an ester bond, an amide bond, or a combination of two or more of such bonds. When two or more compstatin analog moieties are present in a compound, the bonds to the polymer may be the same or different in various embodiments. Doses, regimens, compositions and/or methods can be used to administer long acting compstatin analogs containing compstatin analog moieties attached to GFR by any of such moieties.

[00462] In certain dosage embodiments, administration schedules, compositions, and/or methods described herein can be used to administer LACA that differs in one or more aspects from the LACA used in the clinical studies described herein. For example, such a LACA may contain: (i) a different polymer as the GFR (eg, POS, a PEG-branched polypeptide), (ii) a polymer that has a molecular weight less than or greater than 40 kDa as the GFR; (iii) a different number of compstatin analog fragments (e.g., 1, 3, 4 or more compstatin analog fragments instead of 2 compstatin analog fragments), (iv) one or more compstatin analog fragments containing a sequence that differs from SEQ ID NO: 28, instead of SEQ ID NO: 28 (eg, any of the other compstatin analog fragments described herein); (v) (one or more) compstatin analog fragments extended by another amino acid residue (or an amino acid sequence containing such amino acid residue) that contains a side chain containing a primary or secondary amine instead of lysine, or containing a different reactive functional group, such as sulfhydryl or a sulfhydryl reactive group; (vi) (one or more) compstatin analog fragments fused to the GFR at the N-terminus of the compstatin analog fragment and not at the C-terminus; (vii) a distinct blocking moiety or moieties at the free end of one or more compstatin analog moieties; (viii) any combination of two or more of (i), (ii), (iii), (iv), (v), (vi), (vii) and (vii).

[00463] As mentioned above, in certain dosage aspects, the administration schedules, compositions, and/or methods described herein can be used to administer LACA containing a polymer (e.g., PEG) with a molecular weight of approximately 40 kDa as GFR, e.g., a polymer with a molecular weight of 36 kDa - 44 kDa, 37 kDa - 43 kDa, 38 kDa - 42 kDa or 39 kDa - 41 kDa. However, in some dosage embodiments, administration schedules, compositions, and/or methods can be used to administer LACA containing a polymer whose molecular weight is in a broader range. In certain embodiments of particular interest, the molecular weight of the polymer is not greater than 45 kDa. In certain embodiments, the polymer has a molecular weight of 10 kDa - 45 kDa. In certain embodiments, the polymer has a molecular weight of 20 kDa-45 kDa. In certain embodiments, the polymer has a molecular weight of 30 kDa-40 kDa. In certain embodiments, the polymer has a molecular weight of 30 kDa - 45 kDa. In certain embodiments, the polymer has a molecular weight of 35 kDa-45 kDa. In certain embodiments, the polymer has a molecular weight greater than 45 kDa, for example up to 50 kDa. It is understood that polymer molecular weights may mean the average molecular weight of the polymer molecules in the composition, as discussed herein.

[00464] In certain embodiments, the compositions contain 5% dextrose as a pharmaceutically acceptable carrier. However, any pharmaceutically acceptable carrier may be used.

[00465] In various dose embodiments, administration regimens, compositions, and/or methods described herein can be used to treat any of the complement-mediated disorders mentioned herein. Without limitation, such disorders include, for example, PNH, NZN, myasthenia gravis, aHUS, COPD, idiopathic pulmonary fibrosis, organ transplant rejection (eg, acute or chronic rejection), pemphigoid, and AMD. In certain embodiments, the complement-mediated disorder is a chronic complement-mediated disorder, such as any of the chronic complement-mediated disorders referred to herein. In certain embodiments, the complement-mediated disorder is a Th17-associated disorder, for example, any of the Th17-associated disorders mentioned herein.

[00466] In some aspects, various dosages, administration schedules, compositions, and methods suitable for treating patients by subcutaneous administration of LACA are described herein. In experiments with LACA containing a 40 kDa PEG moiety, it was found that viscosity, not solubility, is the main factor that limits the concentration of a composition containing LACA that can be administered by subcutaneous or/i/t injection in a manner that is appropriate. for physicians and patients in terms of the time and pressure required to deliver a given volume of composition through a needle of a given internal diameter or gauge number. The gauge number describes the outer diameter of the cannula, and a larger gauge number indicates a smaller outer diameter. In general, higher gauge needles may be preferred by patients and clinicians because their use may be (or may be perceived to be) associated with less pain and/or tissue damage compared to lower gauge needles. The inner diameter depends on both the outer diameter and the wall thickness. For standard needles, the higher the gauge number, the smaller the inside diameter (for example, a 27 gauge needle has a larger inside diameter than a 29 gauge needle, which in turn has a larger inside diameter than a 30 gauge needle).

[00467] According to certain embodiments, the composition, for example, a pharmaceutical composition containing LACA, is characterized by a concentration of 150 mg/ml. It was found that a solution of 150 mg/ml LACA containing 40 kDa PEG can be easily delivered using a 27 gauge needle. Accordingly, in some embodiments, a 27 gauge needle is used to administer a dose of LACA containing approximately 40 kDa PEG at concentrations up to 150 mg/ml, e.g., 75 mg/ml - 90 mg/ml, 85 mg/ml - 95 mg/ml, 90 mg/ml - 100 mg/ml, 95 mg/ml - 105 mg/ml 100 mg/ml - 110 mg/ml, 105 mg/ml - 115 mg/ml, 110 mg/ml - 120 mg /ml, 115mg/ml - 125mg/ml, 120mg/ml - 130mg/ml, 125mg/ml - 135mg/ml, 130mg/ml 140mg/ml, 135mg/ml 145mg/ml , 140 mg/ml - 150 mg/ml, for example, 100 mg/ml, 125 mg/ml or 150 mg/ml. In some embodiments, a composition that contains a LACA containing a PEG of approximately 40 kDa is characterized by a concentration of up to 150 mg/mL, e.g., 75 mg/mL - 90 mg/mL, 85 mg/mL - 95 mg/mL, 90 mg /ml - 100 mg / ml, 95 mg / ml - 105 mg / ml, 100 mg / ml - 110 mg / ml, 105 mg / ml - 115 mg / ml, 110 mg / ml - 120 mg / ml, 115 mg / ml - 125 mg / ml, 120 mg / ml - 130 mg / ml, 125 mg / ml - 135 mg / ml, 130 mg / ml 140 mg / ml, 135 mg / ml - 145 mg / ml, 140 mg / ml - 150 mg/ml, for example, 100 mg/ml, 125 mg/ml or 150 mg/ml. In certain embodiments, a composition that contains LACA containing 40 kDa PEG is characterized by a concentration of 150 mg/ml -180 mg/ml. In certain embodiments, a composition that contains a LACA containing a PEG of approximately 40 kDa has a concentration of 180 mg/mL - 200 mg/mL. In certain embodiments, a 27 gauge needle is used to administer a composition that contains LACA containing approximately 40 kDa PEG at a concentration of 150 mg/mL - 160 mg/mL, 160 mg/mL - 170 mg/mL, 170 mg/mL - 180 mg / ml, 180 mg / ml - 190 mg / ml or 190 mg / ml - 200 mg / ml. Of course, instead of a needle with a gauge number of 27 (or higher), a needle with a lower gauge number (for example, gauge 25, 26) can alternatively be used.

[00468] The present description of the invention includes the conclusion that a 200 mg/ml solution of LACA containing 40 kDa PEG can be easily delivered using a 25 gauge needle. In certain embodiments, a composition that contains LACA containing 40 kDa PEG at a concentration of 200 mg/ml or more is administered using a 25 gauge needle or a 26 gauge needle. In certain embodiments, a composition containing LACA at a concentration of 200 mg/ml - 225 mg/ml or 225 mg/ml - 250 mg/ml is administered using a 25 gauge needle or a 26 gauge needle.

[00469] According to some aspects, the present description of the invention presents the specific applicability of certain thin-walled needles for the introduction of LACA according to the present invention. For example, in some embodiments, a thin-walled needle is used for subcutaneous, intramuscular, or intraocular (eg, intravitreal) injection of LACA. Thin-walled needles are characterized by outer diameters identical to the outer diameters of standard needles, but larger inner diameters for a given gauge. For example, a thin-walled needle may have an inside diameter that is the same as a standard gauge needle one to two values lower (e.g., a 29-gauge thin-walled needle may have an inside diameter that is the same as a standard 27 gauge needle). gauge or 28 gauge, but with an outer diameter that is the same as that of a standard 29 gauge needle). Increasing the inside diameter can result in a significant increase in fluid flow at a given pressure and/or a significant decrease in the pressure required to maintain that flow. Less pressure means that less injection force is required to deliver a composition of a given viscosity. Generally, lower injection force facilitates administration and is therefore generally a desirable property. In certain embodiments, the thin-walled needle is characterized by a given inside diameter that is constant along the entire length of the needle. In certain embodiments, the thin-walled needle has an internal diameter that varies along the length of the needle. For example, the diameter may be the same as the diameter of a standard 29 gauge needle at one end of the needle and increase to the diameter of a standard 27 gauge needle at the other end. In certain embodiments, a microconical needle may be used. In certain embodiments, a scalpel-tipped needle may be used. Needle length may vary. In certain embodiments, a short needle may be used, such as a 5 mm or 6 mm needle. According to certain implementation options, you can use a needle, the length of which is from 6 mm to 8 mm or from 8 mm to 12 mm. Suitable needles and syringes are commercially available from, for example, Becton Dickinson and Company (BD), Terumo Corp. etc.

[00470] In certain embodiments, a composition that has a given viscosity and/or concentration can be administered using a thin-walled needle that is one or two gauges larger than the gauge that is preferably used when a standard needle is used to administer the composition of that the same viscosity and/or concentration at a selected flow rate and/or with a selected injection force. For example, in some embodiments, a composition that is preferably administered using a standard 25 gauge needle to achieve the desired flow rate and/or injection force may be administered at the same flow rate and/or injection force using a thin walled 26 gauge needle or 27 gauge. In certain embodiments, a composition that is preferably administered using a standard 27 gauge needle to achieve the desired flow rate and/or injection force may be administered at the same flow rate and/or injection force using a thin walled 28 or 29 gauge needle. . In certain embodiments, a composition that is preferably administered using a standard 29 gauge needle to achieve the desired flow rate and/or injection force may be administered at the same flow rate and/or injection force using a thin walled 30 or 31 gauge needle. . In certain embodiments, a 29-gauge thin-walled needle is used to administer a composition that contains LACA containing a polymer (eg, PEG) with a molecular weight of approximately 40 kDa at a concentration of 150 mg/mL - 160 mg/mL, 160 mg/mL - 170 mg/mL. ml, 170 mg/ml - 180 mg/ml, 180 mg/ml - 190 mg/ml or 190 mg/ml - 200 mg/ml. Of course, instead of a thin-walled needle with a gauge number of 29 (or higher), a thin-walled needle with a smaller gauge number (for example, 27, 28 gauge) can alternatively be used.

[00471] In some aspects, a suitable volume for an individual sc injection may be up to 2 milliliters (ml) - 3 ml. Thus, for example, according to some embodiments, a volume of more than 1.0 ml can be administered by sc injection, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 ml. In certain embodiments, a volume greater than 2.0 ml can be administered by sc injection, such as 2.1, 2.2, 2.3, 2.4, or 2.5 ml. In certain embodiments, a volume greater than 2.5 ml may be administered by sc injection, such as 2.6, 2.7, 2.8, 2.9, or 3.0 ml. In certain embodiments, a volume of 1 ml or less per injection is used, such as 0.5, 0.6, 0.7, 0.8, 0.9, or 1.0 ml. In certain embodiments, the total daily volume administered to the patient is between 1.0 and 2.5 ml, e.g., 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1, 6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4 or 2.5 ml. In some embodiments, the total daily volume administered to the patient is 2.5 to 3.0 ml, such as 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 ml. In some embodiments in which two sc injections are administered, a volume of up to 5 ml may be administered. In certain embodiments, a volume of 1.0-2.5 ml is administered as a single daily injection. In certain embodiments, a volume of 1.0-2.5 ml is administered as two separate injections. In some embodiments, a volume of 2.5-3.0 ml is administered as two separate injections. The volumes administered in two separate injections may be the same or different, as long as they combine to provide a total volume sufficient to administer the desired amount of LACA. In certain embodiments, two separate injections may be administered up to 12 hours apart. However, in some embodiments, two separate injections are administered up to 5, 10, 15, 20, 30, or 60 minutes apart from each other.

[00472] In some aspects, one or more fixed doses may be administered to a variety of patients whose weight and/or body surface area may vary within relatively wide ranges. In certain embodiments, 2, 3 or more different fixed doses may be provided, with different doses being more appropriate for individuals whose weight and/or body surface area are in different ranges, for example.

[00473] In certain embodiments, a fixed dose may be used to administer an amount of a compstatin analog (e.g., LACA) that will achieve a desired level of complement inhibition (e.g., as assessed in a plasma sample) in at least 25%, at least 50% , at least 75%, at least 90%, or at least 95% or more of healthy subjects, and/or at least 25%, at least 50%, at least 75%, at least 90% or at least 95% or more of patients who are in need of treatment for a particular disorder referred to herein. In certain embodiments, a fixed dose may be used to administer an amount that will achieve a desired level of efficacy (e.g., as assessed using an appropriate endpoint) in at least 25%, at least 50%, at least 75%, or at least at least 90% of patients who need treatment for the specific disorder mentioned in this document. In some aspects, the dosage described herein may be particularly suitable for subcutaneous (SC) administration. However, in some embodiments, the doses described herein may be administered to a subject using other parenteral routes of administration, such as intravenous, transdermal, or intramuscular routes, instead of or in addition to the subcutaneous route. Thus, when the specification mentions subcutaneous administration of a particular dose, the specification provides for embodiments in which that dose is administered by a different route of administration (eg, intravenous, transdermal, or intramuscular).

[00474] In certain embodiments, the total daily dose for parenteral, e.g., subcutaneous, administration of LACA comprising two fragments of a compstatin analog and a PEG of approximately 40 kDa is at least 30 mg/day, e.g., at least 45 mg /day, eg at least 90 mg/day, eg at least 150 mg/day, eg at least 180 mg/day. In certain embodiments, the daily dose is 90 mg/day 180 mg/day. In certain embodiments, the daily dose is 180 mg/day - 270 mg/day. In certain embodiments, the dose is, for example, 180 mg/day - 230 mg/day. In certain embodiments, the dose is at least 190 mg/day, eg 190 mg/day - 240 mg/day. In certain embodiments, the dose is at least 200 mg/day, eg 200 mg/day 250 mg/day. In certain embodiments, the dose is at least 210 mg/day, eg 210 mg/day - 260 mg/day. In certain embodiments, the dose is at least 220 mg/day, eg 220 mg/day - 270 mg/day. In certain embodiments, the dose is at least 230 mg/day, eg 230 mg/day - 280 mg/day. In certain specific embodiments, the dose is 180 mg/day, 190 mg/day, 200 mg/day, 205 mg/day, 210 mg/day, 215 mg/day, 220 mg/day, 225 mg/day, 230 mg/day. day, 235 mg/day, 240 mg/day, 245 mg/day, 250 mg/day, 255 mg/day, 260 mg/day, 265 mg/day, or 270 mg/day. In certain embodiments, a dose of 180 mg/day - 270 mg/day is administered in a single daily injection of 1.5 ml - 2.0 ml. In certain embodiments, a dose of 180 mg/day - 270 mg/day is administered in single daily injections up to 2.0 - 2.5 ml. In certain embodiments, a dose of 180 mg/day to 270 mg/day is administered in two daily injections each having a volume of less than 2.0 ml, eg, 1.0 ml to 1.9 ml. For example, in a specific embodiment, a daily dose of 270 mg/day is administered in a single 1.5 ml injection. In another specific embodiment, the daily dose of 270 mg/day is administered in a single 1.8 ml injection. In another specific embodiment, the daily dose of 270 mg/day is administered in two daily injections of 0.9 ml each. In some specific embodiments, the daily dose of 270 mg/day is administered in two daily injections of 0.9 ml of 1.2 ml each, such as 1.0 ml, 1.05 ml, 1.1 ml, 1.15 ml, or 1 .2 ml. In some specific embodiments, a daily dose of 270 mg/day is administered in two daily injections of 1.2 ml - 1.5 ml each, for example, 1.2 ml, 1.25 ml, 1.35 ml, 1.40 ml, 1.45 ml or 1.5 ml. In some specific embodiments, a daily dose of 270 mg/day is administered in two daily injections of 1.5 ml - 1.9 ml each, for example, 1.55 ml, 1.6 ml, 1.65 ml, 1.7 ml, 1.75 ml, 1.8 ml, 1.85 ml or 1.90 ml.

[00475] In certain embodiments, the total daily dose for parenteral, eg, subcutaneous, administration of LACA containing two compstatin analog moieties and approximately 40 kDa PEG is 270 mg/day - 360 mg/day. In certain embodiments, the dose is 270 mg/day - 300 mg/day. In certain embodiments, the dose is 300 mg/day - 330 mg/day. In certain embodiments, the dose is 330 mg/day - 360 mg/day. In certain specific embodiments, the dose is 295 mg/day, 300 mg/day, 305 mg/day, 310 mg/day, 315 mg/day, 320 mg/day, 325 mg/day, 330 mg/day, 335 mg/day. day, 340 mg/day, 345 mg/day, 350 mg/day, 355 mg/day, or 360 mg/day. In certain embodiments, a dose of 270 mg/day - 360 mg/day is administered in a single daily injection of 1.5 ml - 2.0 ml or in a single daily injection of up to 2.0 - 2.5 ml. In certain embodiments, a dose of 270 mg/day - 360 mg/day (e.g., any of the above doses) is administered in two daily injections each less than 2.0 ml in volume, such as between 1.0 ml and 1.9 ml, e.g. 1.0 ml, 1.05 ml, 1.10 ml, 1.15 ml, 1.20 ml, 1.25 ml, 1.30 ml, 1.35 ml, 1.40 ml, 1 .45 ml, 1.50 ml, 1.55 ml, 1.60 ml, 1.65 ml, 1.70 ml, 1.75 ml, 1.80 ml, 1.85 ml, 1.90 ml.

[00476] In certain embodiments, the total daily dose for s.c. administration of LACA containing two compstatin analog and PEG moieties of approximately 40 kDa is at least 360 mg/day, e.g., 360 mg/day - 540 mg/day . In some embodiment, the total daily dose is greater than 360 mg/day, for example up to about 540 mg/day. For example, the total daily dose may be 370 mg/day, 380 mg/day, 390 mg/day, 400 mg/day, 410 mg/day, 420 mg/day, 430 mg/day, 440 mg/day, 450 mg /day, 460mg/day, 470mg/day, 480mg/day, 490mg/day, 500mg/day, 510mg/day, 520mg/day, 530mg/day or 540mg/day. In certain embodiments, a dose of 360 mg/day-540 mg/day is administered in a single daily injection, eg, in a volume of about 2.0-2.5 ml or 2.5 ml-3.0 ml. In certain embodiments, a dose of 360 mg/day up to about 540 mg/day is administered in two daily injections each less than 2.0 ml in volume, such as 1.0 ml to 1.9 ml.

[00477] It is understood that the doses and volumes described herein for subcutaneous (or other parenteral route of administration) administration may be administered using a separate needle and syringe, pen, auto-injector, or other devices known to those skilled in the art. In some embodiments, a syringe pump may be used. In some embodiments, a wearable drug delivery device may be used.

[00478] In some aspects, methods are described herein that include administering a long-acting compstatin analog subcutaneously to a subject at intervals of 2 days or more. As described herein and in PCT/US2016/56144, subcutaneous (SC) administration of 180 mg or more of a compound containing approximately 40 kDa PEG and two fragments of a compstatin analog, referred to herein as LACA-40, per day resulted in pharmacologically significant levels of complement inhibition in a clinical study in healthy subjects (eg, as described in Example 20). Moreover, in clinical studies in subjects suffering from PNH, subcutaneous administration of 180 mg or more of LACA-40 per day resulted in significant changes in blood biomarker levels (eg, as described in examples 21 and 22). Daily s.c. administration of LACA-40 was well tolerated at doses up to 360 mg (the highest dose administered in multiple dose clinical studies as of the filing date of this application). In some aspects, the present invention provides the understanding, at least in part based on data obtained from these clinical studies and pharmacokinetic modeling, that LACA containing approximately 40 kDa PEG and two compstatin analog fragments has a suitable administration profile according to dosing regimens in which doses are administered subcutaneously no more frequently than every two days, for example, every other day, three times a week (sometimes referred to as "three times a week"), once every three days, twice a week (sometimes referred to as " twice a week"), once every four days, once every five days, once every six days, or once a week (sometimes referred to as "once a week" or "weekly"). In some aspects, such administration regimens may provide increased patient convenience compared to, for example, regimens in which LACA is administered daily. In some aspects, such administration regimens may provide a similar or greater mean LACA-40 plasma concentration at steady state compared to daily subcutaneous (SC) administration at a lower dose (e.g., 180 mg to 270 mg daily) and / or a similar or greater minimum (residual) plasma concentration of LACA-40 at equilibrium compared to daily subcutaneous administration at a lower dose (for example, from 180 mg to 270 mg daily), while avoiding excessive fluctuations in plasma concentration, potentially leading to a peak (maximum) plasma concentration that may be undesirably high and/or a residual (trough) plasma concentration that may be undesirably low and may adversely affect efficacy. Without being bound by any theory, it is believed that this fact may be partly due to the fact that LACA-40 is characterized by a relatively slow rate of absorption into the vascular compartment after subcutaneous administration.

[00479] A three times per week schedule is an administration schedule in which a compound, e.g., LACA, e.g., LACA-40, is administered three days a week, with two of those administration days followed by a day on which the compound is not administered, and after the third day of administration, two days follow in which the compound is not administered. For the purpose of describing the introduction schedule, the days of the week can be numbered from Day 1 to Day 7. Obviously, any day of the week can be considered "Day 1". Examples of administration schedules for administration three times a week include dosing on Day 1, Day 3 and Day 5 each week or Day 1, Day 4 and Day 6 each week. For example, assuming that Monday is considered Day 1, an example administration schedule for administration three times a week would be to administer a dose on Monday, Wednesday, and Friday. Many different doses of LACA, for example, LACA-40, can be entered according to the schedule of administration three times a week. In some embodiments, a dose of at least 375 mg, such as 375 mg to 585 mg, is administered subcutaneously three times per week. In some embodiments, a dose of at least 585 mg, such as 585 mg to 885 mg, is administered three times per week. In some embodiments, a dose of 585 mg to 630 mg is administered three times per week. In some embodiments, a dose of at least 630 mg, such as 630 mg to 675 mg, is administered subcutaneously three times per week. In some embodiments, a dose of at least 675 mg, such as 675 mg to 795 mg, is administered three times per week. For example, the dose may be 675 mg to 715 mg, 715 mg to 755 mg, or 755 mg to 795 mg. In some embodiments, a dose of at least 795 mg, such as 795 mg to 885 mg, is administered three times per week. In some embodiments, a dose of at least 840 mg, such as 840 mg to 885 mg or 885 to 930 mg, is administered three times per week. It is also within the scope of the present invention to administer doses of at least 930 mg three times a week, for example doses of 930 mg to 975 mg, 975 mg to 1030 mg, 1030 to 1075 mg, 1075 mg to 1150 mg, 1150 mg to 1240 mg, 1240 mg to 1330 mg, 1330 mg to 1420 mg, 1420 mg to 1510 mg, 1510 mg to 1600 mg, or 1600 mg to 1690 mg.

[00480] The "every other day" schedule, also referred to as the "every other day" or "every two days" schedule, is an administration schedule in which a compound, e.g., LACA, e.g., LACA-40, is administered one once a day every other day, but not on intermediate days. For example, the compound can be administered on Days 1, 3, 5, and 7 of a given week, and then on Days 2, 4, and 6 of the following week in a schedule that is repeated every 2 weeks. In some embodiments, any of the doses described herein in relation to three times per week schedules may be administered on an alternate day schedule. In some embodiments, any of the doses described above for thrice weekly administration can be modified for alternate day administration by reducing the dose by 10% to 15%.

[00481] A twice-weekly schedule is an administration schedule in which a compound, e.g., LACA, e.g., LACA-40, is administered two days per week, with one of these administration days followed by two days in which the compound is not administered. , and after another day of administration, three days follow in which the compound is not administered. Such a dosing schedule may include dosing on Day 1 and Day 4, or on Day 1 and Day 5. For example, assuming Monday is considered "Day 1", an example biweekly dosing schedule would include dosing on Monday and Thursday. A variety of different doses can be administered on a twice-weekly schedule. In some embodiments, a dose of at least 585 mg, such as 585 mg to 900 mg, is administered subcutaneously twice a week. For example, the dose may be 585 mg to 630 mg, 630 mg to 675 mg, 675 mg to 720 mg, 720 mg to 765 mg, 765 mg to 810 mg, 810 mg to 855 mg, or 855 mg up to 900 mg. In some embodiments, a dose of at least 900 mg, such as 900 mg to 945 mg, is administered subcutaneously twice a week. In certain embodiments of particular interest, a dose of at least 945 mg, such as 945 mg to 990 mg, is administered subcutaneously twice a week. In certain embodiments of particular interest, a dose of at least 990 mg, such as 990 mg to 1350 mg, such as 990 mg to 1215 mg, or 1215 mg to 1305 mg, or 1305 mg to 1350 mg, injected subcutaneously twice a week. For example, the dose may be 990 mg to 1035 mg, 1035 mg to 1080 mg, 1080 mg to 1125 mg, 1125 mg to 1170 mg, or 1170 mg to 1215 mg. In some embodiments, a dose of at least 1215 mg, such as 1215 mg to 1260 mg, is administered subcutaneously twice a week. In some embodiments, a dose of 1215 mg to 1395 mg is administered subcutaneously twice a week. In some embodiments, a dose of at least 1260 mg, such as 1260 mg to 1305 mg or 1305 mg to 1350 mg, is administered subcutaneously twice a week. It is also within the scope of the present invention to administer doses of at least 1350 mg twice a week. For example, in certain embodiments, the dose is 1350 mg to 1395 mg, 1395 mg to 1440 mg, 1440 mg to 1485 mg, 1485 mg to 1530 mg, 1530 mg to 1575 mg, 1575 mg to 1620 mg, 1620 mg to 1665 mg, 1665 mg to 1710 mg, 1710 mg to 1755 mg, 1755 mg to 1800 mg, 1800 mg to 1925 mg, 1925 mg to 1970 mg, 1970 mg to 2015 mg, from 2015 mg to 2060 mg, 2060 mg to 2105 mg, 2105 mg to 2150 mg, 2150 mg to 2195 mg, 2195 mg to 2240 mg, 2240 mg to 2285 mg, 2285 mg to 2330 mg, 2330 mg up to 2375 mg, from 2375 mg to 2420 mg, from 2420 mg to 2465 mg or from 2465 mg to 2510 mg.

[00482] The "once every three days" administration schedule is an administration schedule in which a compound, eg, LACA, eg, LACA-40, is administered once every three days, but not on intermediate days. For example, the compound can be administered on Days 1, 4, and 7 of a given week, and then on Days 2 and 5 of the following week, on a schedule that is repeated every 2 weeks. In some embodiments, any of the doses described herein in relation to twice-weekly schedules may be administered on a once-every-three-day schedule. In some embodiments, any of the doses described above for twice weekly administration can be modified for once every three days administration by reducing the dose by 10% to 20% or 20% to 30%.

[00483] The "once every four days" administration schedule is an administration schedule in which a compound, eg, LACA, eg, LACA-40, is administered once every four days, but is not administered on intermediate days. For example, as long as the days are numbered consecutively starting from Day 1, the compound can be entered on Days 1, 5, 9, 13, 17, 21, 25, 29, and so on. In some embodiments, a four day dosing schedule may use any of the doses described herein for twice weekly dosing schedules. In some embodiments, any of the doses described above for twice-weekly administration can be modified to be administered once every four days by increasing the dose by 10% to 20% or 20% to 30%.

[00484] The "once every five days" administration schedule is an administration schedule in which a compound, eg, LACA, eg, LACA-40, is administered once every five days, but is not administered on intermediate days. For example, as long as the days are numbered consecutively starting from Day 1, the compound can be entered on Days 1, 6, 11, 16, 21, 26, 31, and so on. In some embodiments, a five day dosing schedule may use any of the doses described herein for twice weekly dosing schedules. In some embodiments, any of the doses described herein for twice-weekly administration can be modified to be administered once every four days by increasing the dose by 20% to 30%. In some embodiments, any of the doses described herein for twice-weekly administration can be modified to be administered once every four days by increasing the dose by 30% to 40%.

[00485] The "once every six days" administration schedule is an administration schedule in which a compound, eg, LACA, eg, LACA-40, is administered once every five days, but is not administered on intermediate days. For example, as long as the days are numbered consecutively starting from Day 1, the compound can be entered on Days 1, 7, 13, 19, 25, 31, and so on. In some embodiments, a six-day dosing schedule may use any of the doses described herein for weekly dosing schedules. In some embodiments, any of the doses described herein for weekly administration may be modified to be administered once every six days by reducing the dose by 10% to 15%. In some embodiments, any of the doses described herein for weekly administration may be modified to be administered once every six days by reducing the dose by 15% to 20%.

[00486] The once-a-week schedule is an administration schedule in which a compound, e.g., LACA, e.g., LACA-40, is administered one day each week, followed by six days in which the compound is not administered, i. the interval between injections is 7 days. For example, the dose may be administered every Monday. Of course, any day of the week can be chosen as the day on which the dose is administered. A variety of different doses can be administered on a weekly schedule. In some embodiments, a dose of at least 1080 mg, such as 1080 mg to 5040 mg, is administered subcutaneously weekly. In some embodiments, the dose is between 1080 mg and 1440 mg. In some embodiments, the dose is between 1440 mg and 1800 mg. In some embodiments, the dose is between 1800 mg and 2160 mg. In some embodiments of particular interest, the dose is at least 2160 mg. For example, in some embodiments, the dose is between 2160 mg and 2520 mg. Without being bound by any theory, it is believed that a dose of 2160 mg administered subcutaneously weekly is predicted to provide an LACA exposure that is at least as great as that provided by a dose of 270 mg administered daily subcutaneously, while the 2520 mg dose administered subcutaneously on a weekly basis is predicted to provide an effect greater than that provided by the daily subcutaneous dose of 270 mg. In some embodiments, the dose is between 2520 mg and 2880 mg. In some embodiments, the dose is between 2880 mg and 3240 mg. Without being bound by any theory, it is believed that a weekly subcutaneous dose of 2880 mg is predicted to provide an LACA exposure that is at least as great as that provided by a daily subcutaneous dose of 360 mg, while the 3240 mg dose administered subcutaneously on a weekly basis is predicted to provide an effect greater than that provided by the daily subcutaneous dose of 360 mg. In some embodiments, the dose is between 3240 mg and 3600 mg. It is also within the scope of the present invention to administer doses of at least 3600 mg once a week, for example, doses of 3600 mg to 3960 mg, 3960 mg to 4320 mg, up to 4680 mg, from 4680 mg to 5040 mg.

[00487] In some embodiments, the administration schedule is every other day, three times a week, once every three days, twice a week, once every four days, once every five days, once every six days, or weekly provides a residual concentration LACA, e.g. LACA-40, in a plasma at equilibrium that is at least 300 μg/ml, e.g. 300 μg/ml to 325 μg/ml, 325 μg/ml to 350 μg/ml, from 350 µg/ml to 375 µg/ml, 375 µg/ml to 400 µg/ml, or 400 µg/ml to 425 µg/ml. In some embodiments, the dosing schedule is every other day, three times a week, once every three days, twice a week, once every four days, once every five days, once every six days, or weekly to provide a residual concentration of LACA, for example , LACA-40, in plasma at steady state that is at least 425 µg/ml, e.g., 425 µg/ml to 450 µg/ml, 450 µg/ml to 475 µg/ml, 475 µg/ml up to 500 µg/ml, 500 µg/ml to 525 µg/ml, 525 µg/ml to 550 µg/ml, or 550 µg/ml to 575 µg/ml. In some embodiments, the dosing schedule is every other day, three times a week, once every three days, twice a week, once every four days, once every five days, once every six days, or weekly to provide a residual concentration of LACA, for example , LACA-40, in plasma at equilibrium, which is at least 575 μg/ml, for example, from 575 μg/ml to 600 μg/ml, from 600 μg/ml to 625 μg/ml. In some embodiments, the dosing schedule is every other day, three times a week, once every three days, twice a week, once every four days, once every five days, once every six days, or weekly to provide a residual plasma concentration of equilibrium state, which is at least 625 μg/ml, for example, from 625 μg/ml to 650 μg/ml, from 650 μg/ml to 675 μg/ml. In some embodiments, any of the above values can be measured in an individual subject, which may be a healthy subject or may be suffering from a complement-mediated disorder. In some embodiments, the value may be the median or mean value measured in a group of two or more subjects, which may be healthy subjects or may be suffering from a complement-mediated disorder.

[00488] In some embodiments, the ratio of peak plasma concentration to residual plasma concentration (i.e., peak plasma concentration divided by residual plasma concentration), sometimes referred to as the peak/residual action ratio, at equilibrium, which provides an administration scheme every other day, three times a week, once every three days, twice a week, once every four days, once every five days, once every six days or weekly, is not more than 1.5. For example, the peak/residual action ratio may be 1.05 to 1.1, 1.1 to 1.2, 1.2 to 1.3, 1.3 to 1.4, or 1.4 to 1.5. In some embodiments, any of the above relationships can be measured in an individual subject, which may be a healthy subject or may be suffering from a complement-mediated disorder. In some embodiments, the indicated value may be the median or mean value measured in a group of two or more subjects, which may be healthy subjects or may be suffering from a complement-mediated disorder.

[00489] In some embodiments, LACA, for example, LACA-40, can be administered at a concentration of 25 mg/ml to 50 mg/ml, for example, when administered according to the administration schedule every other day, three times a week, two times a week, once every three, four, five or six days, or once a week. In some embodiments, such administration is carried out using a pump, such as a syringe pump. In some embodiments, such administration is via a wearable delivery device. In some embodiments, LACA, such as LACA-40, can be administered at a concentration of 50 mg/mL to 75 mg/mL, such as when administered according to a schedule of administration every other day, three times a week, twice a week, once a day. three, four, five or six days or once a week. In some embodiments, such administration is carried out using a pump, such as a syringe pump. In some embodiments, such administration is via a wearable delivery device. In some embodiments, LACA, such as LACA-40, can be administered at a concentration of 75 mg/mL to 100 mg/mL, such as when administered according to a schedule of administration every other day, three times a week, twice a week, once a day. three, four, five or six days or once a week. In some embodiments, such administration is carried out using a pump, such as a syringe pump. In some embodiments, such administration is via a wearable delivery device. In some embodiments, LACA, such as LACA-40, can be administered at a concentration of 100 mg/mL to 150 mg/mL, such as when administered according to a schedule of administration every other day, three times a week, twice a week, once a day. three, four, five, or six days, or once a week In some embodiments, such administration is via a pump, such as a syringe pump. In some embodiments, such administration is via a wearable delivery device. Thus, in various embodiments, LACA, for example, LACA-40, can be administered according to any of the administration schedules described herein, for example, subcutaneously at intervals of 2 days or more, for example, according to the administration schedule every other day, three times a week , once every three days, twice a week, once every four days, once every five days, once every six days, or weekly, using any suitable delivery device, such as obtained from any of the device vendors described in this document.

[00490] The concentration of the LACA composition and the volume administered can be selected to provide the desired dose for administration weekly, twice a week, or three times a week (or for administration every other day, once every three days, once every four days, once in five days, once every six days), for example, any of the doses described herein. In some embodiments, LACA, such as LACA-40, is administered in a volume of 2 ml to 50 ml, such as 5 ml to 10 ml, 10 ml to 15 ml, 15 ml to 20 ml, 20 ml to 25 ml. ml, 25 ml to 30 ml, 30 ml to 35 ml, 35 ml to 40 ml, 40 ml to 45 ml, or 45 ml to 50 ml. It should be understood that such volumes can be used for administration according to other administration schedules described herein, for example, for more frequent administration, for example, administration daily, according to certain embodiments. For example, in some embodiments, a dose of 270 mg is administered daily sc in a 10 ml volume at a concentration of 27 mg/ml or in a 5 ml volume at a concentration of 54 mg/ml. In some embodiments, a dose of 360 mg is administered daily sc in a 10 ml volume at a concentration of 36 mg/ml or in a 5 ml volume at a concentration of 72 mg/ml. Typically, the volume does not exceed 50 ml, although in certain embodiments, larger volumes may be used, for example, up to 100 ml or more, especially if doses greater than 5000 mg are administered.

[00491] It should be understood that a subject treated according to any of the administration regimens described herein (e.g., administration regimens including administration daily, every other day, three times a week, once every three days, twice a week , once every four days, once every five days, once every six days, or weekly) may suffer from any of the complement-mediated disorders described herein. For example, in some embodiments, the subject suffers from a complement-mediated hemolytic anemia, such as PNH, or from a hemolytic anemia, which includes antibodies that bind to RBCs (red blood cells) and lead to complement-mediated lysis, such as autoimmune hemolytic anemia with warm agglutinin syndrome or cold agglutinin disease. In some embodiments, the subject suffers from myasthenia gravis. In some embodiments, the subject is suffering from optic neuromyelitis. In some embodiments, the subject suffers from a complement-mediated disorder that affects the kidneys, such as C3 glomerulopathy or IgA nephropathy.

[00492] In some aspects, a dosage unit of LACA, for example, LACA-40, may contain any of the amounts described herein as suitable for an administration schedule daily, every other day, three times a week, two times a week, once every three , four, five or six days or once a week. In some embodiments, single dosage unit a is greater than 540 mg, such as at least 541 mg, such as at least 545 mg. The dosage unit may contain an amount within any of the ranges described herein, or any specific value within the specified range. In some embodiments, a container, cartridge, syringe, or drug delivery device containing said dosage unit is described herein.

[00493] In certain embodiments, treatment with a compstatin analog, e.g., LACA, by subcutaneous administration or other parenteral route of administration, e.g., according to any of the administration schedules and/or using any of the doses or administration devices described herein, can be continue for any period of time, such as indefinitely. In certain embodiments, the patient may be treated with a short course of treatment (eg, for up to 1 week, 2 weeks, or up to 1, 2, 3, 4, 5, 6 months). In certain embodiments, such a short course of treatment may be sufficient to modify the course of a disorder (e.g., PNH, AIHA (autoimmune hemolytic anemia), NZN, myasthenia gravis, COPD, asthma, idiopathic pulmonary fibrosis, vasculitis, pemphigoid, nephropathy) so that when treatment was interrupted, a long-term remission was maintained (for example, which is confirmed by the absence of exacerbations or relapses), i.e. LACA may be washed out of the body between treatments. In certain embodiments, the subject may receive short course treatment once or twice a year, every two years, and so on. In certain embodiments, a short course of treatment may modify the course of a disorder (e.g., PNH, AIHA, NZN, myasthenia gravis, COPD, asthma, idiopathic pulmonary fibrosis, vasculitis, pemphigoid, nephropathy) such that long-term remission or cure is achieved without further treatment. Without being bound by a particular theory, it is believed that a short course of treatment with a compstatin analogue, such as LACA, may be sufficient to disrupt the cycle that maintains an immune response against cells or tissues (eg, their components such as proteins or lipids) that are targeted. immune system attacks in complement-mediated disorders. In certain embodiments, for example, a short course of LACA treatment may be sufficient to disrupt the cycle that maintains an immune response against cells or tissues in the bone marrow (eg, against hematopoietic stem cells) (eg, in a patient with PNH), in the nervous system ( e.g., against nerve cells, glial cells) (e.g., in a patient with ONH or myasthenia gravis), in the circulatory system (e.g., against endothelial cells, red blood cells) (e.g., in a patient with vasculitis), in the respiratory system (e.g., in a patient with COPD, asthma, or idiopathic pulmonary fibrosis), in the integumentary system (eg, in a patient with pemphigoid).

[00494] In some aspects, various dosages, administration schedules, compositions, and methods of use are described herein for the treatment of a patient in need of treatment for a complement-mediated ocular disorder that affects the posterior segment of the eye, e.g., the retina, by intravitreal (and/in /r) introduction of LACA. In certain embodiments, the ocular disorder is AMD, for example, advanced AMD (geographic atrophy (GA) or neovascular AMD), diabetic retinopathy, glaucoma, or uveitis.

[00495] In certain embodiments, the dose for intravitreal injection of LACA containing two compstatin analogue fragments and a PEG of approximately 40 kDa is 5 mg to 20 mg. In certain embodiments, the dose is 10 mg. In certain embodiments, the dose is 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, or 20 mg. In certain specific embodiments, the dose is 15 mg. In certain embodiments, any of the aforementioned doses are administered by intravitreal injection in a volume of 90 to 110 microliters, for example, in a volume of 100 microliters. In certain embodiments, any of the above doses is administered by intravitreal injection using a 27, 28, 29, or 30 gauge needle. In phase 1b clinical studies, doses up to 20 mg (ie 5 mg, 10 mg and 20 mg) in 100 microliter volumes, administered by intravitreal injection using a 29 gauge needle, were found to be well tolerated by patients with AMD. In certain embodiments, a dosage and needle gauge combination is selected that provides for administration by i/v/t injection in 10 seconds or less. In certain embodiments, a dose-needle combination is used that delivers a dose by i/v/t injection in 5-6 seconds or less. A dose of 15 mg in a volume of 100 microliters (150 mg/mL) has been found to result in a composition that has a favorable viscosity for administration by intravitreal injection in 5-6 seconds or less through a 27 gauge thin walled needle.

[00496] In certain embodiments, the dose of the LACA-containing composition is administered by i/v/t injection with a volume greater than 100 microliters. For example, a volume of 100-110 microliters, 110-125 microliters, or 125-150 microliters can be used. Such larger volumes may allow for the administration of a higher dose than a 100 microliter injection volume without increasing the time required for dose delivery and/or without the need for a smaller gauge (larger diameter) needle. Such an increase in the total dose may be proportional to the increase in volume. For example, a 50% increase in dose volume would provide a 50% increase in the amount of LACA administered without increasing the time required for dose delivery and/or without the need for a smaller gauge needle.

[00497] In certain embodiments, the dose of the LACA-containing composition is administered by ophthalmic administration (e.g., i/v/t injection) once a day, once every 6 weeks, or once every 2 months (i.e., every other month ). In certain embodiments, the dose of the LACA-containing composition is administered by i/v injection once every 3 months, once every 4 months, once every 5 months, or once every 6 months, or less frequently, such as once every 9 months, once a year). Thus, in some embodiments, a patient may receive 1 to 6 injections per year, typically at approximately equal time intervals. In certain embodiments, the patient is initially treated with monthly injections (e.g., for the first 3-6 months or the first 6-12 months) followed by less frequent administration (e.g., every 1, 3, 4, 5, or 6 months or less often, e.g. once every 9 months, once a year).

[00498] In certain embodiments, LACA treatment by i/v/t injection according to any of the aforementioned administration schedules can be continued indefinitely. In certain embodiments, the patient may be treated with a short course of treatment (eg, 1, 2, 3, 4, 5, or 6 i/v/t injections). In certain embodiments, a short course of treatment may be sufficient to halt or substantially halt the progression of a disorder (eg, AMD, eg, GA, or early or intermediate AMD), such that additional treatment is not required. For example, a short course of LACA treatment may be sufficient to disrupt the cycle that maintains an immune response against the retina or retinal pigment epithelium.

[00499] In certain embodiments, LACA is administered using a syringe with one or more design features that reduce friction and/or force required during injection, such as a relatively short barrel and/or a relatively larger plunger size.

[00500] In experiments conducted using LACA containing PEG of various molecular weights, it was determined that a solution of LACA containing a lower molecular weight PEG (for example, 10 kDa - 30 kDa) has a reduced viscosity compared to LACA containing 40 kDa PEG , at the same concentration in mg/ml. A lower viscosity may favor the use of a higher gauge needle (eg 29 gauge instead of 27 gauge). In certain embodiments, LACA containing a polymer, e.g., PEG, with a molecular weight of less than 40 kDa, can be administered using a needle with a smaller inner diameter and/or a larger gauge number than LACA containing a polymer, e.g., PEG, with a molecular weight of 40 kDa or more. In certain embodiments, a LACA containing a polymer, such as PEG, with a molecular weight of less than 40 kDa can be administered at a concentration of 80 mg/mL to 150 mg/mL, such as about 100 mg/mL or about 125 mg/mL. In certain embodiments, LACA containing a polymer, such as PEG, with a molecular weight of less than 40 kDa can be administered at a concentration of 150 mg/mL to 250 mg/mL. In certain embodiments, a LACA containing a polymer, e.g., PEG, with a molecular weight of less than 40 kDa (e.g., 10 kDa - 35 kDa) can be administered at a higher concentration than 250 mg/ml, for example, up to 300 mg/ml, 300 mg / ml - 400 mg / ml, 400 mg / ml - 500 mg / ml or more.

[00501] In certain embodiments, a LACA comprising a GFR (e.g., PEG, POS, polypeptide, or other polymer) with a molecular weight of less than about 40 kDa (e.g., 10 kDa - 35 kDa) and a specified number of compstatin analog fragments (e.g., 1, 2, 3, 4) can be administered at approximately the same molar dose as any of the amounts described above for LACA containing approximately 40 kDa GFR, resulting in a lower dose by weight still containing approximately the same number of compstatin analog fragments. The introduction of a smaller total amount of connection per dose (by weight) may allow to reduce the volume of the dose. Reducing the volume of dose administration may allow for a reduction in the number of injections (eg, one daily injection instead of two daily injections in some embodiments) and/or may provide a reduction in injection time (the length of time an individual dose is administered). A lower total dose (by weight) may allow the use of a lower concentration (by weight) of the compound in a given volume, resulting in reduced viscosity and allowing the use of needles with a smaller internal diameter and/or a shorter injection time. In certain embodiments, a LACA containing a GFR (e.g., PEG, POS, polypeptide, or other polymer) with a molecular weight of less than about 40 kDa (e.g., 10 kDa 30 kDa) and a specified number of compstatin analog fragments (e.g., 1, 2, 3, 4 ) can be administered at a higher molar dose than LACA containing approximately 40 kDa GFR, resulting in the administration of more compstatin analogue fragments per dose. In certain embodiments, the molar dose can be up to 1.2, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5 times greater depending on, for example, the molecular weight of GFR.

[00502] In certain embodiments, a LACA comprising a GFR (e.g., PEG, POS, polypeptide, or other polymer) with a molecular weight of less than about 40 kDa (e.g., 10 kDa - 35 kDa) and a specified number of compstatin analog fragments (e.g., 1, 2 , 3, 4) can be administered at a lower dose (by weight) per unit time (eg, lower daily dose, lower weekly dose, lower monthly dose, etc.), at about the same dose per unit time or at a higher dose per unit time than that described above for LACA containing a GFR of approximately 40 kDa. In certain embodiments, it is contemplated that the dose (by weight) of LACA containing a GFR with a molecular weight of less than about 40 kDa (e.g.; 10 kDa - 30 kDa) may be lower or higher by up to a factor of up to about 3 (e.g., about 1, 1, 1.2, 1.5, 2.0, 2.5, 3.0) compared with a daily dose of LACA containing 40 kDa GFR.

[00503] The present invention encompasses the administration of a compstatin analog in combination with adjunctive therapy. Such additional therapy may include the introduction of any(s) agent(s) used in the art or potentially suitable for the treatment of a subject suffering from the specified disease. For example, in some embodiments of LACA, a cell-reactive compstatin analog or targeted compstatin analog is administered in combination with a C5 inhibitor (e.g., eculizumab or any of the other C5 inhibitors mentioned herein or known in the art) to a patient, e.g., a patient with PNH or any of the other complement-mediated disorders mentioned herein. In certain embodiments of LACA, a cell-reactive compstatin analog or targeted compstatin analog is administered in combination with an anti-vascular endothelial growth factor (VEGF) agent to a subject with wet AMD. Anti-VEGF agents include antibodies that bind to VEGF, such as ranibizumab (Lucentis) and bevacizumab (Avastin), polypeptides containing the soluble portion of the VEGF receptor, such as aflibercept (Eylea, also known as VEGF-Trap).

[00504] If two or more therapeutic agents (e.g., compounds or compositions) are used or administered “in combination” with each other, according to various embodiments of the present invention, they can be administered at the same time, over overlapping time periods, or sequentially (for example, can be separated in time by a period of up to 2 weeks or more, for example, separated in time by a period of up to 4, 6, 8 or 12 weeks). They can be introduced in the same way or in different ways. In some embodiments, said compounds or compositions are administered at 48 hour intervals. In some embodiments, the compstatin analogue may be administered before or after the administration of the additional compound(s), e.g., close enough in time such that said compstatin analogue and the additional compound(s) are(are) present in appropriate amounts. in the body at least once. In some embodiments, said compounds or compositions are administered close enough in time such that no more than 90% of the previously administered composition is metabolized to inactive metabolites or eliminated, e.g., by elimination from the body, by the time the second compound or composition is administered.

[00505] In some embodiments, a composition is administered comprising both a cell-reactive compstatin analog or a long-acting or targeted compstatin analog and an additional compound or compounds.

[00506] In certain embodiments, a subject who is being treated or is to be treated with a compstatin analog using, for example, inhalation or parenteral administration by, for example, subcutaneous, intramuscular, or intravenous injection, is vaccinated against one or more pathogens. For example, the subject may receive a vaccine against Neisseria meningitidis, Haemophilus influenzae and/or Streptococcus pneumoniae. In certain embodiments, the subject is vaccinated against all three of these organisms. In certain embodiments, the subject may receive the vaccine at least 1, 2, 3, 4, 5, or 6 weeks prior to the first dose of the compstatin analog. If necessary, the subject may receive one or more additional doses of the vaccine.

[00507] In certain embodiments, a subject who is being treated or should be being treated with a compstatin analogue using, for example, inhalation administration or parenteral administration by, for example, subcutaneous, intramuscular, or intravenous injection, may receive an antibiotic that is expected to be effective in preventing or limiting infection caused by one or more pathogens such as Neisseria meningitidis, Haemophilus influenzae and/or Streptococcus pneumoniae. In certain embodiments, the subject may receive an antibiotic prophylactically. Prophylactic administration may begin before the subject has received the first dose of the compstatin analogue, or at any time thereafter. Suitable antibiotics are known to those of ordinary skill in the art. For example

Illustrative examples

[00508] Example 1: Preparation of pegylated compstatin analogs substantially retaining complement inhibitory activity

[00509] A compstatin analogue with the amino acid sequence of the compstatin analogue of SEQ ID NO: 28 but containing an AEEAc-Lys fragment located C-terminally from the Thr residue of SEQ ID NO: 28 was synthesized for subsequent conjugation of an NHS ester-activated PEG with an amino group side chain of Lys. This compound was synthesized using standard methods. Briefly, amino acids (including AEEAc) were obtained as Fmoc-protected amino acids, characterized in that the α-amino group of each amino acid is protected with Fmoc. The functional groups of the side chains are also blocked with various suitable protecting groups. The synthesis was carried out by the solid phase method described by Merrifield (J. Amer. Chem. Soc. 85, 2149 (1963)). The chains were collected on the solid phase, at the end of which the N-terminus was acetylated; the peptide was then cleaved from the solid phase, simultaneously deprotected by acidolysis using TEA, and amidated. The linear peptide was then oxidized and purified. Compstatin analog obtained is represented by the following: Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-AEEAc-Lys-M/2 (SEQ ID NO : 51), abbreviated as CA28-AEEAc-Lys. Note that, for brevity, the N-terminal acetyl group and C-terminal amino groups are not indicated in the abbreviation. 30 kDa and 40 kDa monofunctional linear activated PEG NHS ester (NOF America Corp. White Plains, NY, Cat. No. SUNBRIGHT® ME-300GS and Cat. No. SUNBRIGHT® ME-400GS), respectively, coupled to the lysine side chains of CA28-AEEAc-Lys to give the following long acting compstatin analogs: CA28-AEEAc-Lys-(PEG30k) and CA28-AEEAc-Lys-(PEG40K) and purified. Note that the number after the term "PEG" and before the letter "k" is the molecular weight of the indicated PEG fragment in kilodaltons, and "k" is short for "kDa"). CA28-AEEAc-Lys-(PEG30k) is also referred to as CA28-1. CA28-AEEAc-Lys-(PEG40K) is also referred to as CA28-2.

[00510] The inhibitory activity of synthesized compounds was assessed by measuring the effect of said compounds on complement activation in the classical way using a standard complement inhibition assay. This protocol measures C3b deposition in ELISA format. C3b deposition monitored in this manner is mediated by classically activated complement. Briefly, 96-well plates are coated with BSA. Human plasma, chicken ovalbumin (OVA), anti-OVA polyclonal antibody, and test compound (referred to as "drug") are added, incubated, and then hP-conjugated anti-human C3 antibody is added. After additional incubation, the substrate is added and the signal is recorded. The details of the protocol are given below:

[00511] Classical Complement Pathway Inhibition Assay Protocol

Materials:

• 96-well plate (Polystyrene plate, Thermo Scientific, 9205)

• Chicken ovalbumin (Sigma A5503-5G)

• Rabbit anti-chicken ovalbumin antibody (Abeam abl221)

• Blocking buffer (Startingblock buffer, Thermo Scientific 37538)

• Veronal buffer (concentration 5X, Lonza 12-624E)

• Human plasma (collected supplemented with lepirudin at a final concentration of 50 µg/mL)

• HRP-conjugated goat anti-human C3, (MP Biomedicals, 55237)

• Wash buffer Tween-20 (0.05% Tween 20-FSB buffer)

• TMB (Peroxidase Substrate, BD 555214) - 1:1 mixture of BD 51-2607KC and 51-2606KC.

• 1M H ₂ SO ₄

Protocol:

1. Add 100 µl/well 1% chicken ovalbumin (in PBS)

2. Incubate overnight at 4°C or room temperature for 12 hours.

3. Remove by shaking and tapping the plate.

4. Block by adding 200 µl blocking buffer

5. Incubate for 1 hour at room temperature

6. Remove by shaking and tapping the plate

7. Add 100 µl anti-chicken ovalbumin polyclonal antibody 1:1000 dilution in blocking buffer

8. Incubate for 1 hour at room temperature

9. Wash twice with wash buffer

10. Add 50 µl of VB ⁺⁺ to wells #2-12

11. Add 100 µl of drug at initial dilution (2X in VB ⁺⁺ ) to well 1.

12. Prepare serial dilutions (1:2) of the indicated drug from wells 1-10 as follows:

a. Take 50 µl of solution from the original well

b. Add the specified solution to the next well

With. Mix by repeated pipetting

d. Repeat until hole #10

Note: Remove 50 µl from well #10 and discard.

13. Add 50 µl of plasma at a 2X dilution (1:37.5 dilution of original plasma) to wells 111

14. Incubate for 1 hour

15. Rinse with wash buffer

16. Add 100 µl 1/1000 dilution of hP anti-C3 antibody in blocking buffer

17. Incubate for 1 hour

18. Rinse with wash buffer

19. Add 100 µl TMB to all wells

20. Incubate for 5 10 min in the dark

21. Add 50 µl 1M H2SO4*

22. Read plate at 450 nm

VB ⁺⁺

Formula:

barbital 5 mM NaCl 72.5 mm MgCl ₂ 0.5 mm _CaCl2 0.15 mM pH 7.4

Initial solutions:

To obtain 50 ml working buffer:

• Weigh out 210 mg NaCl

• Add 10 ml 5X VB

• Add 100 µl CaCl ₂ (500X)

• Add 250 µl MgCl (200X)

• Make up to 50 ml with H ₂ O

• Adjust pH to 7.4

[00512] Data was analyzed using GraphPad Prism5 software. The data sets for each experiment were normalized for percent activation compared to a control with 100% activation corresponding to the well to which no compound was added. Drug concentration values (X values) were logarithmically converted and percent activation (Pa) (Y values) were converted to percent inhibition (Pi) using the following formula: Pi=100-Pa (Yi=100-Ya). Build a graph of the percentage of suppression of the concentration of the drug; to approximate the resulting dataset, a sigmoidal dose-response function [Y=Bottom+(Top-Bottom)/(1+10 ((Log EC-X)))] was used. IC ₅₀ values were obtained using the fitting function parameters.

[00513] The results are shown in FIG. 1, the IC ₅₀ values are shown in Table 2 (in Example 2). According to the data, CA28-1 and CA28-2 exhibited an activity of approximately 30% of the activity of CA28 in terms of moles.

[00514] Example 2: Preparation of Long-acting Compstatin Analogs Demonstrating Increased Molar Activity

[00515] 8 arm 40 kDa activated NHS ester PEG (NOF America Corp. White Plains, NY, cat. no. SUNBRIGHT® HGEO-400GS; chemical formula: hexaglycerol octa(succinimidyloxyglutaryl)polyoxyethylene) was attached to the lysine side chain of CA28-AEEAc-Lys to give the following long-acting compstatin analogue: (CA28-AEEAc) ₈ -PEG40K, also referred to as CA28-3. Complement-suppressing activity of CA28-3 was investigated using the assay described in Example 1. The results are plotted in FIG. 1, and the IC50 value is shown in Table 1, both as a function of CA28 concentration. The concentration of CA28 was calculated using the extinction coefficient of CA28 at 283 nm (10208.14 l⋅mol ^-1 ⋅cm ^-1 ). Based on another analysis (UV absorbance versus material mass and elemental CHN analysis), it was concluded that there are 7.5 CA28 moieties per CA28-3 molecule. Thus, the activity of CA28-3 on a mole basis is 7.5 times higher than that shown in FIG. 1 and shown in Table 2. Thus, the IC50 value in Table 2 is 7.5 times higher than the actual IC50 value of CA28-3 in terms of moles. When calculated, the IC50 value of CA28-3 in terms of moles is approximately 0.26 (less than that of the parent compound CA28). In FIG. 2 shows the percentage of complement inhibitory activity of CA28 and long-acting compstatin analogs CA28-2 and CA28-3 as a function of CA28-3 concentration (µM), i.e. the activity of CA28-3 was corrected for the fact that the specified compound contains 7.5 CA28 fragments. On a mole basis, the complement inhibitory activity of CA28-3 is higher than that of CA28.

[00516] The solubility of CA28-1, CD28-2 and CA28-3 in water in the presence or absence of a variety of buffers and/or excipients was observed to be higher than the solubility of the parent compound CA28.

[00517] Example 3: Long-acting compstatin analogues demonstrating a significant increase in plasma half-life and Cmax

[00518] This example describes the determination of the pharmacokinetic parameters of the long acting compstatin analogs CA28-2 and CA28-3 after administration to cynomolgus monkeys.

[00519] Dosing and sample collection

[00520] CA28-2 and CA28-3 were administered at time 0 by intravenous injection to female cynomolgus monkeys (three per group, age 2-5 years, 2.9-3.5 kg). Compounds were administered at 50 mg/kg in 5% dextrose in water at a concentration of 25 mg/ml. Blood samples (~1 ml each) were collected from the femoral vein at the following time points: pre-dose, 5 min, 15 min, 30 min, 1 hour (h), 4 h, 8 h, 24 h, 48 hours, 96 hours (4 days) and 192 hours (8 days) after dosing. Samples were collected by direct venipuncture, placed in a red cap serum tube containing no anticoagulant, and kept at room temperature for at least 30 minutes. Blood samples were centrifuged at 4°C at 3000×g for 5 minutes. Throughout the processing, the samples were kept cool. Serum samples were collected after centrifugation and placed in sample tubes. The samples were stored in a freezer set to maintain temperatures from -60°C to -80°C. All animals showed normal activity throughout the study. No compound-related abnormalities were noted in the animals during the course of the study.

[00521] Sample analysis. Plasma samples obtained as described above were analyzed by LC/MS/MS using the following methods for determining the concentration of the compound: 50 μl of the sample was mixed with an internal standard (CA28-AEEAc-Arg), then 100 μl of 1 M NH ₄ OAc was added, pH 3.5 with HOAc and stirred. Then 250 μl of acetonitrile was added and mixed. The sample was centrifuged, the supernatant was poured into another tube and dried. The sample was redissolved and injected into the LC/MS/MS system. Mobile phase A was provided with 5 mM NH ₄ OAc with 0.1% FA, Mobile phase B was provided (ACN:50 mM NH ₄ OAc) in a 90:10 ratio with 0.1% FA. An Intrada WP-RP 2×150 mm, 3 µm LC column was used. Quantification was performed on an Applied Biosystems API-4000 triple quadrupole mass spectrometer in positive ion mode. Collision-induced dissociation (CDI) was used to fragment the compound in the source of the mass spectrometer, ions with m/z 144 were selected by mass in Q1, fragmented, ions with m/z 77 were selected by mass in Q3 and detected. Data were processed using Analyst 1.4.2 software.

[00522] Results. Serum concentrations in μg/ml of CA28-2 and CA28-3 at each time point are shown in Table 3 below. Data are presented for each of the 3 monkeys treated with the indicated compound. Means and standard deviations are easy to calculate. Data for all animals are in good agreement. For CA28, data are already available from a previous study in which CA28 was intravenously administered to cynomolgus monkeys. In this study, CA28 was determined in samples using HPLC.

[00523] The results for each compound were averaged and the resulting graph is shown in FIG. 3. A significant increase in half-life and Cmax compared to CA28 was observed for both CA28-2 and CA28-3. The terminal half-life for both CA28-2 and CA28-3 was approximately 4-4.5 days. Based on these data, intravenous administration at approximately 1-2 week intervals is expected to provide consistent compound levels and effectively suppress complement activation in human subjects, although dosing intervals may be both shorter and longer.

[00524] Example 4: Long-acting compstatin analog containing HSA as a clearance-reducing moiety

[00525] Human serum albumin (HSA) side chain lysines were converted to thiols using 2-iminothiolane and reacted with a compstatin analog containing maleimide as a reactive functional group: Ac-Ile-Cys*-Val-Trp(1-Me )-Gln-Asp-Trp-Gly-Ala-His-Arg-Cys*-Thr-AEEAc-Lys-(C(=O)-(CH ₂ ) ₅ -Mal)-NH ₂ (SEQ ID NO: 68) . The complement inhibitory activity (FIG. 4) of the resulting long acting compstatin analog (CA28-4) was tested in vitro as described in Example 1, and the pharmacokinetic properties were tested in vivo as described in Example 3. The pharmacokinetic parameters of CA28-4 after administration to cynomolgus monkeys were determined as described in the previous example. The results are shown in FIG. 5 (together with results for CA28, CA28-1, CA28-2 and CA28-3). PK data for CA28-4 are presented in Table 4.

[00526] Example 5: Synthesis and activity of PEG-based compstatin analogues using various activated NHS-PEGs

[00527] A compstatin analog having a compstatin analog amino acid sequence corresponding to the sequence of SEQ ID NO: 28, but containing an AEEAc-Lys fragment located C-terminally from the Thr residue corresponding to the sequence of SEQ ID NO: 28, for the purposes of subsequent conjugation of activated NHS-ester PEGs with the amino group of the Lys side chain were synthesized as described in example 1. The resulting analog of compstatin is represented by the following: Ac-Ile-Cys*-Val-(1Me)Trp-Gln-Asp-Trp-Gly-Ala-His-Arg- Cys*-Thr-AEEAc-Lys-NH ₂ (SEQ ID NO: 51), abbreviated as CA28-AEEAc-Lys. Monofunctional linear activated 40 kDa monomethoxy NHS ester/carbonate PEGs differing in NHS carboxylate addition chemistry (NOF America Corp. White Plains, NY, cat. no. SUNBRIGHT® ME-400CS, SUNBRIGHT® ME-400GS, SUNBRIGHT® ME-400HS, SUNBRIGHT® ME-400TS) were linked to the CA28-AEEAc-Lys lysine side chain by an amide bond. (The Lys residue is Lys 15 because the AEEAc linker contains an amino acid residue.) All compounds were N-terminally acetylated, C-terminally amidated, and disulfide cyclized between Cys2 and Cys12. (Acetylation, amidation, and cyclization were performed to PEG coupling.) Compounds were obtained as trifluoroacetate salts and purified. These compounds are shown in the following table (table 5). The designation letters CS, GS, HS and TS correspond to different linker moieties between the PEG moiety and the NHS moiety as described in more detail in Table 5. It will be understood that the different names and designations for each compound can be used interchangeably. Note that CA28-2 (see example 1) is identical to CA28-2GS.

[00528] Compounds were analyzed by reverse phase HPLC. In FIG. 6 shows a representative chromatogram of one of these compounds. A VariTide RFC column was used. Eluent A: 0.1% TFA in water; Eluent B: 0.1% TFA in 50% ACN/40% water. The flow rate was 1,000 ml/min, with a gradient from 0% B to 100% B over 40 minutes. The peak with a retention time of 33.68 minutes corresponds to the pegylated compound and has a relative area of 96.50%.

[00529] The inhibitory activity of these compounds was assessed by measuring the effect of these compounds on complement activation via the classical pathway using a standard complement inhibition assay as described in Example 1. The results are plotted in FIG. 7. The results shown are a combination of the results of two separate experiments. The compounds showed substantially similar complement inhibitory activity.

[00530] Example 6: Synthesis and activity of PEG-based bifunctionalized analogs of compstatin

[00531] Bifunctional linear activated monomethoxy NHS ester/carbonate 40 kDa PEGs differing in NHS carboxylate addition chemistry were obtained from NOF America Corp. (White Plains, NY). Activated PEGs were linked to the lysine side chain of CA28-AEEAc-Lys by an amide bond such that two CA28-AEEAc-Lys fragments were linked to each PEG chain. All compounds were acetylated at the N-terminus and amidated at the C-terminus of the CA28-AEEAc-Lys fragments and cyclized with a disulfide bond between Cys2 and Cysl2. (Acetylation, amidation and cyclization were performed prior to coupling with PEG.) Compounds were obtained as acetate salts and purified. These compounds are represented by the compounds shown in the following table (table 6).

[00532]

[00533] The inhibitory activity of CA28-2GS-BF was assessed by measuring the effect of said compound on complement activation via the classical pathway using a standard complement inhibition assay as described in Example 1 and analyzed as described in Example 1. The results are plotted in FIG. 8. As described above, CA28-2GS-BF contains two compstatin analog moieties per molecule. Although CA28-2GS-BF has lower activity per compstatin analog moiety than the activity of the individual CA28 molecule, the molar activity of the two compounds is virtually identical over a wide range of concentrations.

[00534] Example 7: Subcutaneous administration of a PEG-based bifunctionalized analogue of compstatin

[00535] This example describes the determination of the pharmacokinetic parameters of the long-acting compstatin analog CA28-2GS-BF after administration to cynomolgus monkeys either by a single intravenous (IV) injection or by multiple (once a day) subcutaneous injections for seven days.

[00536] Dosing and Sampling CA28-2GS was administered at time O by intravenous injection or multiple subcutaneous injections (daily for seven days) to male cynomolgus monkeys. This study used six previously treated male cynomolgus monkeys, aged 15 years, whose body weight ranged from 4.6 to 5.3 kg. At the start of the seven-day study, the animals were healthy. The study was not blind. Animals had free access to water and were fed commercial feed twice daily prior to the start of the study. Feed was provided to the animals using a standard design device (SOP) prior to the start of the study. Animals were not limited in food. Animals were dosed intravenously and subcutaneously at time 0 on the corresponding day. For subcutaneous injection, a 22 gauge needle was used. The compound was administered at a concentration of 50 μg/kg in 5% dextrose in water at a concentration of 25 mg/ml. Blood samples (~1 ml each) were collected from the femoral vein at the following time points: Day 1: before drug administration, 5 min, 15 min, 30 min, 1 hour (h), 4 h, 8 h. Days 2 9:0 min. Day 16: Last sample based on day 1 dose. Each blood sample (˜1.0 ml) was collected from the monkey's femoral or saphenous vein by direct venipuncture and placed in a red-capped serum tube containing no anticoagulant and kept at room temperature for at least 30 minutes. Blood samples were centrifuged at 4°C at 3000×g for 5 minutes. The samples were kept refrigerated throughout the processing. Serum samples were collected after centrifugation and placed in sample tubes. The samples were stored in a freezer to maintain a temperature of -60°C - -80°C. Serum samples and remnants of dosing solutions for analysis were transported frozen on dry ice.

[00537] Each site of subcutaneous injection was examined to establish the rate of absorption of the injection volume, as well as to determine whether after the introduction of the composition, the seal remains or it completely resolves. Dosing sites were examined at each sampling and during the daytime for 27 days. All doses were absorbed throughout the study. Based on observations, doses were absorbed within 15 minutes of administration. All animals showed normal activity throughout the study. Animals showed no junction-related abnormalities throughout the study.

[00538] Sample analysis. Plasma samples obtained as described above were analyzed using LC/MS/MS enhanced with CID (collision-induced dissociation) in a manner similar to that described in Example 3.

[00539] Results. Serum concentration of CA28-2GS-BF versus time when administered intravenously or subcutaneously as described above is plotted in FIG. 9. Data points correspond to all detected CA28 pegylated compounds. The data for CA28 shown in FIG. 9 are retrospective data from a previous study in which CA28 was intravenously administered to cynomolgus monkeys. In this study, CA28 was detected in samples using HPLC.

[00540] A peak serum concentration of 500 μg/ml (11 μM) was achieved by subcutaneous administration of CA28-2GS-BF. The terminal half-life of CA28-2GS-BF was approximately 5 days when administered either iv or subcutaneously. The results are summarized in the tables below:

[00541] Example 8: Inhibition of complement-mediated lysis of red blood cells from patients with PNH

[00542] A modified Ham test was performed to measure the ability of compstatin analogues to inhibit complement-mediated lysis of red blood cells from PNH patients in vitro. Complement was activated with acidified magnesium-supplemented serum to lyse PNH-affected red cells. Incubation was carried out for 90 minutes. Data were recorded using flow cytometry of PNH-affected red cells using standard markers. Heat-inactivated serum (not causing hemolysis) was used as a control. Acidified serum without the addition of a complement inhibitor causes maximum lysis. The experiment was performed with serial twofold dilutions of compstatin analogs CA28, CA28-2, CA28-2CS, CA28-2CS-BD, CA28-2GS, CA28-2GS-BF, CA28-2HS, CA28-2HS-BF, CA28-2TS, CA28- 2GS-BF and CA28-3. The concentration of each compound required to completely block hemolysis in vitro was determined. Also, red blood cells were stained for C3 fragment deposition using anti-C3 polyclonal antibodies that do not contain any bridges to cause agglutination (e.g., Ab4214 or Ab14396, both commercially available as FITC conjugates from Abcam, Cambridge, UK), to measure the ability of these compounds to inhibit the deposition of C3 fragments on red blood cells in PNH. The results were compared with those obtained for eculizumab using the same assays.

[00543] Example 9: Long-acting compstatin analogues in patients with PNH

[00544] The cohort of subjects diagnosed with PNH was divided into 4 groups. Subjects in Groups 1 and 2 were treated with intravenous CA28-2 or CA28-3, respectively, at a dose of 5 mg/kg and 20 mg/kg, at intervals of 1 and 2 weeks. Treatment was optionally initiated at shorter intervals and then reduced with maintenance therapy. Subjects in Group 3 received eculizumab treatment according to the recommended dosing regimen. Group 4 served as control (no complement inhibitor therapy). Changes over time were monitored for intravascular hemolysis (based on measurements of LDH and/or cCK(51)Cr labeling), reticulocytosis (indicator of anemia), hematocrit, hemoglobin concentration in the blood, erythrocyte opsonization (deposition of C3 activation products, such as C3b, on erythrocytes, which can be detected using flow cytometry), symptoms of PNH, need for transfusion, thromboembolism, hemolysis-related nitric oxide depletion, rates of pulmonary hypertension, quality of life, and survival. Results were compared between groups and retrospective data for control patients with PNH obtained from clinical trials of eculizumab. Improvement in persistent anemia (as evidenced, for example, by decreased reticulocytosis, decreased signs of hemolysis, increased hematocrit, increased hemoglobin levels), improved quality of life, reduced symptoms of PNH, reduced need for transfusions, decreased incidence of thromboembolism, decreased hemolysis-related nitric oxide depletion , reduction in rates of pulmonary hypertension, improved quality of life and/or improved survival in subjects receiving CA28-2 (Group 1) or CA28-3 (Group 2) compared with subjects in group 4, is indicative of efficacy.

[00545] Example 10: Long-acting compstatin analogues in patients with PNH

[00546] Example 9 was repeated with the difference that the subjects were individuals with PNH who remained transfusion dependent and/or whose hemoglobin content continued to remain below the cut-off value (e.g., 9.0 g/dL) despite treatment with eculizumab. The results were compared between groups.

[00547] Example 11: Long-acting compstatin analogs in patients with aHUS

[00548] The cohort of subjects diagnosed with aHUS was divided into 4 groups. Subjects in Groups 1 and 2 were treated with intravenous CA28-2 or CA28-3, respectively, at a dose of 5 mg/kg to 20 mg/kg, at intervals of 1 to 2 weeks. Treatment was optionally initiated at shorter intervals and then reduced with maintenance therapy. Subjects in Group 3 receive eculizumab treatment according to the recommended dosing regimen. Changes over time were monitored for intravascular hemolysis (based on LDH measurements), erythrocyte opsonization (deposits of C3 activation products such as C3b on erythrocytes), aHUS symptoms, renal function, need for plasma exchange or dialysis, quality of life and survival. Results were compared between groups and retrospective data for control patients with aHUS obtained from clinical trials of eculizumab. Reducing signs of hemolysis, improving quality of life, reducing symptoms of aHUS, reducing the need for plasma exchange or dialysis, improving quality of life and/or improving survival in subjects receiving CA28-2 or CA28-3 compared to subjects in group 4 is indicative of efficacy.

[00549] Example 12: Examples 811 were repeated using CA28-2GS-BF, CA28-2HS, CA28-2HS-BF, CA28-2TS and CA28-2GS-TS-BF.

[00550] Example 14: Examples 9-12 were repeated using CA28-2GS-BF, CA28-2HS, CA28-2HS-BF, CA28-2TS and CA28-2GS-TS-BF administered daily by subcutaneous injection.

[00551] Example 14: Examples 8-11 were repeated using additional long-acting compstatin analogues.

[00552] Example 15: Examples 8-11 were repeated using cell-reactive analogs of compstatin.

[00553] Example 16: Activation of Complement Inhibitory Activity of a Long-Acting Compstatin Analog

[00554] CA28 and CA28-AEEAc-Lys were synthesized as described above. CA28-2TS-BF was synthesized using a reactive bifunctional TS-type PEG for NHS-carboxylate addition chemistry, which was coupled to two molecules of CA28-AEEAc-Lys via the primary amine of the lysine side chain. Complement activation inhibitory activity of CA28 and CA28-2TS-BF was assessed by measuring the effect of these compounds on complement activation via the classical pathway and via the alternative pathway using standard complement inhibition assays. The Classical Pathway Activation Assay protocol is described in Example 1. The Alternate Pathway Activation Protocol also measures C3b deposition in an ELISA format and is described below. The deposition of C3b, which is monitored using this method, is achieved by complement activated through an alternative pathway of lipopolysaccharide (LPS). Briefly, 96-well plates are coated with LPS. Test compound (referred to as "drug") is added, followed by plasma or serum as a source of complement, and incubated. hP-conjugated anti-human C3 antibody is then added. After additional incubation, the substrate is added and the signal is recorded. A detailed description of the protocol is given below:

[00555] ELISA-based Complement Activation Via Alternative Pathway Assay

Materials:

• 96-well ELISA plate (Corning 3590)

• LPS from Salmonella typhosa - Sigma L7136 (40 µg/ml in PBS)

• BSA 1% in PBS - Calbiochem #126626, dilution 1/30

• Veronal buffer +10 mM MgCl ₂ +10 mM EGTC (VB-Mg EGTC)

• Human plasma (collected supplemented with lepirudin at a final concentration of 5 µg/mL)

• hR-conjugated antibodies against human C3 (polyclonal antibodies to C3-HRP Ab, Cappel 55237)

• Wash buffer Tween-20 (0.05% in PBS)

• TMB (Peroxidase Substrate) - 1:1 mix of BD 51-2607KC and 51-2606KC.

• 3M H ₂ SO ₄

• Microplate Reader

Protocol:

1. Add 50 µl/well of LPS at a concentration of 40 µg/ml (in PBS)

2. Incubate for 2 hours at room temperature

3. Remove by shaking and tapping the plate.

4. Block by adding 200 µl 1% BSA/PBS

5. Incubate for 1 hour at room temperature

6. Remove by shaking and tapping the plate

7. Add 50 µl of VB-Mg EGTA to wells #2-12

8. Add 100 µl of first dilution drug (2x in VB-Mg EGTA) to well 1.

9. Prepare serial dilutions (1:2) of the drug from wells 1-10 as follows:

a. Take 50 µl of solution from the original well

b. Add the specified solution to the next well

c. Mix by repeated pipetting

d. Repeat until hole #10

Note: Remove 50 µl from well #10 and discard

10. Add 50 µl of 2x plasma dilution to wells 111

11. Incubate for 1 hour

12. Wash twice with wash buffer

13. Add 50 µl of C3-HRP Ab diluted 1/1000 in 1% BSA/PBS

14. Incubate for 1 hour

15. Add 100 µl of TMB to all wells

16. Incubate for 30 min.

17. Add 50 µl 3M H ₂ SO ₄

18. Read plate at 450 nm

Formula VB Mg EGTC

Barbital 5 mM NaCl 72.5 mm MgC12 10 mM EGTC 10 mM pH 7.3-7.4

Initial solutions:

To obtain 20 ml working buffer:

• Weigh out 84 mg NaCl

• Add 4 ml 5X VB

• Add 2 ml EDTA 10X

• Add 2 ml MgCl 10X

• Make up to 20 ml with H ₂ O

• Adjust pH to 7.4

[00556] Results

[00557] FIG. 10(A) shows the percent inhibition of the classical complement pathway by the inhibitory activity of CA28 and CA28-2TS-BF as a function of the molar concentrations of these compounds. In FIG. 10(B) shows the percent inhibition of the alternative pathway of complement activation by the inhibitory activity of CA28 and CA28-2TS-BF as a function of the molar concentrations of these compounds. Raw data are shown in Table 9 below (4 replicates for each condition). Based on the inhibition curves shown in the figures and the underlying data, the complement inhibitory activity of CA28-2TS-BF is at least equal to that of CA28 on a molar basis within the experimental error of this assay. These results further confirm the suitability of the long acting compstatin analogs described herein, for example, for therapeutic use.

[00558]

[00559] Example 17: Pharmacokinetic properties of a long-acting compstatin analogue when administered intravenously or subcutaneously

[00560] This example describes the determination of the pharmacokinetic parameters of the long-acting compstatin analog CA28-2TS-BF after administration to cynomolgus monkeys by a single intravenous (IV) injection, a single subcutaneous injection, or subcutaneous injection once a day for seven days. CA28-2TS-BF was synthesized using a reactive bifunctional TS-type PEG for NHS-carboxylate addition chemistry, which was coupled to two molecules of CA28-AEEAc-Lys via the primary amine of the lysine side chain.

[00561] Dosing and sample collection

[00562] CA28-2TS-BF was administered to cynomolgus monkeys at time 0 by intravenous injection into the saphenous vein, or by single subcutaneous injection or repeated subcutaneous injections (once a day for seven days). This study used six previously treated female cynomolgus macaques (three per group), aged 25 years, whose weight ranged from 2.6 to 3.9 kg. The animals were healthy at the start of the study. The study was not blind. Animals had free access to water and were fed commercial feed twice daily prior to the start of the study. Feed was provided to the animals using a standard design device (SOP) prior to the start of the study. Animals were not limited in food. Animals were administered 7 μg/kg intravenously or subcutaneously at time 0 on the respective day(s). The concentration of the compound to be dosed in solution was 3.5 mg/mL for IV administration and 25 mg/mL for subcutaneous administration. The dose volume was 2 ml/kg for intravenous administration and 0.28 ml/kg for subcutaneous administration. For subcutaneous injection, a 23G3/4 gauge needle was used. The compound was administered in 5% dextrose in water.

[00563] Blood samples (-0.5-1 ml) were collected from the femoral vein at the following time points: Day 1: pre-drug, 5 min, 15 min, 30 min, 1 hour (h), 4 h , 8 hours Days 2 - 9: 0 min. Day 15: Last sample based on day 1 dose. Each blood sample was collected from a monkey's femoral vein by direct venipuncture and placed in a red cap serum tube containing no anticoagulant and kept at room temperature for at least 30 minutes. Blood samples were centrifuged at 4°C at 3000 xg for 5 minutes. The samples were kept refrigerated throughout the processing. Serum samples were collected after centrifugation and placed in sample tubes. The samples were stored in a freezer to maintain temperatures from -60°C to -80°C. Serum samples and remnants of dosing solutions for analysis were transported frozen on dry ice.

[00564] Each site of subcutaneous injection was examined to establish the rate of absorption of the injection volume, as well as to determine whether after the introduction of the composition, the seal remains or it completely resolves. Dosing sites in subcutaneously injected animals were examined on the evening of each dosing day. No induration was observed at the dosing sites and the dose was completely absorbed by this time as judged by visual inspection. All animals were examined twice a day; the animals showed normal activity throughout the study. Animals showed no junction-related abnormalities throughout the study.

[00565] Sample analysis. Plasma samples obtained as described above were analyzed using LC/MS/MS enhanced with CID (collision-induced dissociation) in a manner similar to that described in Example 3.

[00566] Results.

[00567] Serum concentrations of CA28-2TS-BF versus time when administered intravenously or subcutaneously as described above are plotted in FIG. 11. Data points correspond to all detected CA28 pegylated compounds. The data for CA28 shown in FIG. 11 are retrospective data from a previous study in which CA28 was intravenously administered to cynomolgus monkeys. In this study, CA28 was detected in samples using HPLC/MS.

[00568] The peak serum concentration of approximately 500 μg/ml was achieved by subcutaneous administration of CA28-2TS-BF once a day for 7 days. The terminal half-life of CA28-2TS-BF was approximately 8 days when given either iv or a single subcutaneous injection. Raw data are shown in Tables 10(A) (IV) and 10(B) (SC) below. (In FIG. 11 and Tables 10(A) and 10(B), the day of dosing is considered day 0).

[00569] As noted above, CA28-2TS-BF was synthesized using a reactive, bifunctional TS-type PEG to form a carbamate after reaction with the primary amine of lysine. CA28-2GS-BF was synthesized using a reactive bifunctional GS-type PEG for NHS-carboxylate addition chemistry to form an amide after reaction with the primary lysine amine. Said compound also contains an ester bond not found in CA28-2TS-BF. It should be noted that the terminal half-life of approximately 8 days achieved with CA28-2TS-BF in this experiment was markedly longer than that of CA28-2GS-BF, which was found to have a half-life of approximately 5 days in a similar experiment. experiment (see example 8).

[00570] Example 18: Compstatin analogs inhibit C3 deposition on red blood cells of PNH patients and protect against complement-mediated lysis

[00571] A modified Ham test was performed to evaluate the ability of compstatin analogues to protect affected PNH cCC from complement-mediated lysis. CCC cells from a PNH patient exposed to acidified human serum (as a source of complement components) and magnesium (Mg ²⁺ , required to activate the alternative pathway) in the absence of complement inhibitors or in the presence of varying amounts of compstatin analogues CA28 or CA28-2GS-BF . Exposure to heat-inactivated human serum was used as a control corresponding to the absence of significant complement-mediated lysis, since complement is inactivated by heat. Exposure to acidified human serum and magnesium (Mg ²⁺ ) in the absence of complement inhibitors (panel labeled "Mg ²⁺ ") was used as a control corresponding to maximum lysis.

[00572] After incubation, cells were stained with antibodies against CD59 and C3d. The level of CD59 allowed the classification of affected PNH cCC as type I, type II, or type III cells. Staining for C3d, the product of C3 activation and cleavage, was used as a marker for C3 and deposition of C3 activation products (load). Flow cytometry analysis was performed to assess CD59 and C3d on cKC surfaces and quantify the percentage of type I, type II, and type III cells present in various samples.

[00573] The results of a dilution experiment demonstrating the effect of different concentrations of CA28 on C3 deposition and percentage of cells are shown in FIG. 12(A). The results of a dilution experiment demonstrating the effect of different concentrations of CA28-2GS-BF on C3 deposition and percentage of cells are shown in FIG. 12(B). Quantitative results are presented in Table 11 below. Type I cells (highlighted in orange in Fig. 12) contain normal levels of CD59. Type III cells (highlighted in blue in FIG. 12) contain essentially no detectable CD59. These cells are very sensitive to complement-mediated lysis. CD59 levels in type II cells (shown in purple in FIG. 12) are reduced compared to normal or type I cells and are moderately susceptible to complement-mediated lysis. When complement is activated, type III cells rapidly undergo lysis. A decrease or absence of lysis may be indicated by the presence of an increased number of type III cells, as indicated by a higher percentage of type III cells in the panel where there is no lysis compared to the panel (in the presence of Mg ²⁺ (maximum lysis)), as in FIG. 12(A) and 12(B). In other words, the positive control contains relatively fewer type III cells than the negative control. Type II cells may eventually be lysed in the presence of activated complement, but before that they may accumulate a significant amount of C3 activation products, such as C3d. Decrease or absence of lysis may be indicated by increased levels of C3 or C3 activation products on type II cells, as indicated by comparing the level of C3d on type II cells in the panel where lysis is absent, with the level of C3d on type II cells in the panel with maximum lysis on both figures, fig. 12(A) and FIG. 12(B). In other words, cells in the panel with maximum lysis have more C3d than cells in the panel with no lysis. Type I cells contain functional CD59; thus, they deactivate the convertase and therefore do not accumulate as much C3d as type II cells. At the same time, the amount of C3d that they accumulate can be used as a surrogate indicator of the level of lysis of more vulnerable cells (types II and III). Accordingly, a reduced amount of C3d on type I cells indicates protection from lysis. A shift in the relative percentages of type I, II and III cells relative to the percentages in the control panels with maximum lysis (Mg ²⁺ ) towards the percentages in the control panels where lysis is absent (heat-inactivated serum) indicates protection from complement-mediated lysis. These percentages are shown in the table below. The column labeled "%C'3" in Table 11 refers to the percentage of cells counted as "positive" for C3 and C3 activation products ("C3 load"). As seen in FIG. 12(A) and (B) and from Table 11, CA28 and CA28-2GS-BF at tested concentrations showed similar protection against lysis of PNH-affected red cells with virtually no C3 loading of PNH-affected red cells at compound concentrations of 100 μg/mL or greater. high concentrations. Note that the percentages of type III, II and I cells in the presence of 100 μg/ml or more of the compstatin analog were essentially the same as the percentages in the control with no lysis, indicating complete protection from complement-mediated lysis as assessed by this assay. Concentrations below 100 µg/mL but above 60 µg/mL, such as at least 70 µg/mL, at least 80 mg/mL, or at least 90 µg/mL but below 100 µg/mL have not been tested in the specified experiment, but may also provide significant protection. 100 μg/ml CA28-2GS-BF corresponds to a concentration of approximately 2.5 μM, which is readily achievable in vivo as described herein.

[00574]

[00575] Example 19: Effect of Compstatin Analog and Soliris on C3 Deposition on Red Blood Cells of a PNH Patient

[00576] An experiment similar to that described in Example 18 was conducted to further demonstrate the protective effect of the compstatin analogue CA28-2GS-BF and compare it to that of the anti-C5 antibody Soliris. A modified Ham assay according to Example 18 was performed using PNH-affected cCC incubated in the presence of activated complement either in the absence of a complement inhibitor (left panel) or in the presence of Soliris (middle panel) or CA28-2GS-BF (50 μg/mL) (right panel). panel). After antibody staining with antibodies against CD59 and C3d, flow cytometry was performed. The results are shown in FIG. 13. In this figure, quadrant 1 (Q1) and quadrant 3 (Q3) correspond to type III cells. Quadrant 2 (Q2) and quadrant 4 (Q4) correspond to type I and II cells. Q1 and Q2 correspond to cells with a significant and abnormally high amount of C3 activation product deposition (eg, C3d). Q3 and Q4 correspond to cells without significant deposition of C3d or with slightly elevated levels (right side of Q4), but less than cells in Q2. The percentage of cells in different quadrants is shown below each panel in FIG. 13 and in table 12 below.

[00577]

[00578] As can be seen, in the absence of inhibitors, the vast majority of cells are located in Q4 (type I or type II, with low levels of deposition of C3 activation products). Type III cells had to be mostly lysed, so their percentage (Q1 and Q3) is low. Cells in Q2 that accumulate C3 deposition products eventually undergo lysis, so their numbers remain relatively small. In the presence of eculizumab, type III cells are protected from lysis, at least initially, but accumulate C3 activation products (eg, C3d), as evidenced by the high percentage of cells in Q1 compared to the panel without inhibitor (36.79% vs. 0.09%) . The relative abundance of Q2+Q4 cells (types I and II) is lower as a result of increased survival of type III cells. However, it is clear that significant deposition of C3 activation products (eg C3d) occurs on type III cells, which may eventually lead to lysis or excretion (in vivo). CA28-2GS-BF-treated (right panel) afflicted PNH cCC essentially did not show C3d deposition, whether type I, II, or III, in contrast to the results obtained with eculizumab. The percentage of cells in Q1 and Q2 is negligible. There is a dramatic increase in the percentage of type III cells (61.55%) compared to results obtained without inhibitor or with eculizumab, indicating (along with the absence of C3d deposition) increased protection from lysis by CA28-2GS-BF.

[00579] Example 20: Phase 1 clinical studies of a long-acting compstatin analogue in healthy subjects

[00580] Two randomized, double-blind, placebo-controlled phase 1 clinical trials of a long-acting compstatin analog containing a linear 40 kDa PEG and two compstatin analog fragments (each attached to each end of the linear PEG), each containing a peptide with the amino acid sequence of SEQ ID NO: 28, a C-terminally extended fragment containing AEEAc-Lys to attach a PEG fragment, were initiated to evaluate safety, tolerability, pharmacokinetics and pharmacodynamics: a study of single ascending doses (SAD) and multiple incremental doses ( multiple ascending doses, MAD). In examples 20-26, this compound is referred to for convenience as LACA-40. In a single incremental dose study, healthy subjects were randomized to one of six cohorts with doses ranging from 45 mg to 1440 mg (45, 90, 180, 360, 720, or 1440 mg) in 5% dextrose. LACA-40 was administered by subcutaneous injection on the first day of the study followed by 29 or 43 days of observation, depending on the dose level administered. Each cohort included 4 subjects who received drug and 1 or 2 subjects who received placebo. In a multiple incremental dose study, LACA-40 was administered by subcutaneous injection to healthy subjects daily for 28 consecutive days followed by 56 observation days after the last dose. Subjects were included in one of four cohorts with doses ranging from 30 mg to 270 mg/day (30, 90, 180, or 270 mg/day). Each cohort included 4 subjects who received drug and 1 subject who received placebo. Safety was assessed through intensive clinical observation. Serial blood sampling was performed to determine serum LACA-40 concentrations. Blood samples were also taken to determine the appropriate complement activity markers (C3, CH50 and AP50). In the latter cohorts, additional relevant FD markers were measured (pharmacodynamics; intact C3, iC3b, C3a, C4a and C5a). Prior to multiple doses, subjects received vaccinations against Neisseria meningitides, Streptococcus pneumoniae, and Haemophilus influenzas type B (Hib).

[00581] Results

[00582] When a total of 24 healthy subjects received single doses of LACA-40 at doses up to 1440 mg in a phase 1 single incremental dose study and a total of 16 healthy subjects received multiple doses of LACA-40 for 28 consecutive days at doses up to 270 mg/day in a multiple escalating dose study, 11 healthy subjects received single or multiple placebo doses in the studies, analysis found that LACA-40 was well tolerated in both studies with no serious adverse events or post-treatment adverse events resulting in to discontinuation of the drug study, or reported severe adverse events. In addition, in each study, there were no safety signals of clinical relevance when examining laboratory data, vital signs, physical examinations, or electrocardiography results.

[00583] The pharmacokinetics of LACA-40 were consistent with predictions based on preclinical data, with little variability between patients.

[00584] In a multiple escalating dose study, plasma concentrations of LACA-40 were found to increase linearly with dose, reaching peak concentrations on days 14 to 28. Serum concentrations after 28 days of daily dosing were close to steady state.

[00585] Both studies found a dose-dependent increase in C3 levels, indicating that LACA-40 binds to C3.

[00586] In a single incremental dose study, a decrease in alternative pathway mediated hemolytic activity (AP50) was observed after a single dose of 1440 mg of LACA-40.

[00587] In the third cohort of the multiple escalating dose study of LACA-40 at a dose level of 180 mg/day, a reduction in ex vivo serum-induced hemolysis was observed as early as seven days after the start of treatment, which continued throughout treatment and peaked at more than 80% in two of the four subjects and more than 60% in the other two subjects (Fig. 14). In the fourth cohort study of multiple incremental doses of LACA-40 at a dose level of 270 mg/day, a decrease in serum-induced hemolysis ex vivo was observed as early as seven days after the start of treatment, which continued during treatment and reached a maximum of more than 80% in three of four subjects. The result for the fourth subject is an outlier: he experienced a decrease of approximately 40% from baseline.

[00588] Percent inhibition of serum-induced hemolysis ex vivo was determined based on a standard rabbit erythrocyte hemolysis assay when complement is activated by the alternative pathway (AR). Rabbit erythrocytes are spontaneous activators of human AR. The assay exploits the fact that when rabbit erythrocytes are incubated in serum with the addition of EGTA to chelate Ca ²⁺ (to inhibit complement activation by the classical and lectin pathways), AP convertase is formed leading to C3 activation and subsequent erythrocyte lysis, which can be detected. by detecting free hemoglobin by spectrophotometry. It should be noted that this assay is likely to underestimate the actual percentage of hemolysis inhibition because there is no serum in the negative control sample, while serum is present in samples from subjects and contributes to baseline uptake in these samples.

[00589] It should be noted that in clinical studies of eculizumab, a complement inhibitor that binds to C5, in patients with PNH, 80% inhibition of serum-induced hemolysis ex vivo (using serum from patients with PNH) has been shown to be of significant benefit in treatment of PNH (Hillmen, P., et al., N Engi J Med 2004; 350:552-9). The present example, in this regard, confirms that pharmacologically significant levels of complement inhibition were achieved at least at a dose of LACA-40 180 mg/day. This example also confirms that the pharmacological doses of LACA-40 were safe and well tolerated, that the pharmacokinetic/pharmacodynamic profile of LACA-40 (PK/PD) supports daily sc administration, and that the daily doses of LACA-40 180 mg and 270 mg significantly reduced hemolytic activity as early as seven days after the start of dosing, and this inhibition was maintained throughout the entire period of dosing.

[00590] In one aspect, PK data from these studies was used to develop a PK/PD model that can be used to facilitate dose selection in patients with PNH or other patients who are administered LACA-40.

[00591] The present disclosure provides that lower doses of LACA-40 may also be effective in PNH. For example, the ex vivo serum-induced hemolysis assay only measures MAC-induced lysis, which reflects intravascular hemolysis. LACA-40 (and certain other compounds described herein) protect cells from MAC as well as from opsonization by C3 fragments, such as C3b, which cause extravascular hemolysis and potential dysfunction that eculizumab does not alleviate. Therefore, without wishing to be bound by theory, the present disclosure provides that in the treatment of PNH, according to some embodiments, efficacy can be achieved even at doses that are lower than required to inhibit 80% of ex vivo serum-induced hemolysis, at least partly due to inhibition of extravascular hemolysis.

[00592] Thus, among other things, this example demonstrates that subcutaneous dosing (e.g., daily subcutaneous dosing over an appropriate period of time, e.g., at least one day, 2 days, 3 days, 4 days , 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks or more) of a long-acting compstatin analog containing a linear 40 kDa PEG and two fragments of a compstatin analog, effective effects can be achieved. The present example specifically demonstrates the effective effects that were achieved with a daily dose of 180 mg/day, and specifically provides that both higher and lower doses may well be suitable in appropriate circumstances. The present example also specifically demonstrates the effective effects that were achieved with a daily dose of 270 mg/day, and specifically provides that both higher and lower doses may well be suitable in appropriate circumstances.

[00593] Moreover, in light of the specific demonstration of the effective results that have been achieved with the proposed LACA-40 administration regimens, the present example confirms the specific utility of long acting compstatin analogs, and in particular analogs containing at least two compstatin analog moieties, and /or analogues containing a PEG fragment with a molecular weight of approximately 40 kDa (for example, in the range from approximately 10 kDa to approximately 50 kDa, specifically including a molecular weight of approximately 20 kDa, 30 kDa, 40 kDa, etc.), in such schemes .

[00594] Alternatively or additionally, this example demonstrates the specific applicability of long-acting compstatin analogs that have a total molecular weight of no more than about 50 kDa in the administration regimens described herein (see above).

[00595] Example 21: Phase Ib LACA-40 Clinical Study in PNH Subjects

[00596] A phase Ib clinical study of single and multiple incremental doses of LACA-40 was initiated to evaluate the safety, tolerability, PK and PD of LACA-40 in combination with eculizumab (Soliris) in adult patients with PNH. In this clinical study, subcutaneous doses of LACA-40 in 5% dextrose were administered to patients with PNH, all of whom were receiving concomitant eculizumab therapy. To participate in the study, patients must be at least 18 years of age, weigh over 55 kg, have been treated with eculizumab for at least 3 months, have a hemoglobin level of <10 g/dl at screening, or have received at least one blood transfusion within 12 months prior to screening, characterized by a platelet count >30,000/mm ³ and an absolute neutrophil count >500 cells/µl. Prior to dosing, all subjects were started on prophylactic oral antibiotics and vaccinated against Neisseria meningitides. Subjects in cohorts 3 and 4 were also vaccinated against Streptococcus pneumoniae and Haemophilus influenzae type B (Hib).

[00597] Each of the first two cohorts consisted of two patients who received a single dose of LACA-40 followed by at least 28 days of observation. If after this observation period it was concluded that the single dose was well tolerated, the patient then received a daily subcutaneous dose regimen of LACA-40 for an additional 28 consecutive days. The third and fourth cohorts consisted of two and six patients, respectively, who received subcutaneous doses of LACA-40 daily for 28 consecutive days. The studied doses were as follows:

Cohort 1: Single sc dose of 25 mg LACA-40 and repeated sc doses of 5 mg/day

Cohort 2: Single sc dose of 50 mg LACA-40 and repeated sc doses of 30 mg/day

Cohort 3: Repeated sc doses of 180 mg/day LACA-40

Cohort 4: Repeated sc doses of 270 mg/day LACA-40

[00598] Safety was assessed through clinical observation and all drugs were administered by qualified nurses at the subjects' home or clinic. Serial blood serum samples were taken to determine the concentrations of LACA-40. Pharmacodynamic (PD) activity and efficacy signals were assessed, including levels of lactate dehydrogenase (LDH), hemoglobin, PNH cCC clone distribution, blood transfusion requirements, complement levels, C3 fragment deposition on cCC and reticulocytes.

[00599] Results

[00600] When the dosing of the first three cohorts was completed, the analysis showed that LACA-40 was well tolerated; one severe adverse event was reported, which was assessed as probably not associated with the introduction of LACA-40.

[00601] When dosing two subjects was completed with the pharmacologically active dose of 180 mg, both subjects showed clinical improvement and corresponding changes in blood biomarkers. Hemoglobin levels increased in both subjects during the first two weeks of treatment and remained stable until the end of treatment on day 28. LDH levels were stable at ~1.5x ULN (upper limit of normal) in one subject and decreased from ~1.5x ULN to within normal range in another subject. The cCC ratio of PNH type III (CD59-negative) approximately doubled in both subjects, rising from 22.3% to 52% and from 32.5% to 62.5% from day 1 to day 29, respectively. RCC transfusion requirements also decreased during the dosing period compared to recent history data. No treatment-related serious adverse events were reported, and there were no treatment-related adverse events leading to treatment discontinuation. Proceeded to dosing cohort 4 (270 mg daily).

[00602] When dosing to three subjects for 28 days of dosing with sc LACA-40 270 mg/day was completed, all three subjects experienced similar clinical improvement associated with corresponding changes in blood biomarkers. Hb levels increased, LDH levels decreased, reticulocyte levels decreased, and the percentage of PNH type III cCC increased. Based on a risk/benefit profile assessment, a protocol amendment was approved to allow continuous dosing in the 270 mg/day cohort for an additional 56 days for a total of 84 days of treatment in addition to ongoing eculizumab therapy. None of the three subjects required a cCC transfusion since the start of LACA-40 treatment.

[00603] The data confirm that pharmacological doses of LACA-40 are safe and well tolerated in subjects with PNH, and daily SC dosing of LACA-40 provides sustained inhibition of hemolytic activity in patients with PNH. The data also confirm that inhibition of C3 provides clinical benefit in subjects with PNH who have a suboptimal response to anti-C5 treatment (eg, treatment with eculizumab).

[00604] Thus, the present disclosure provides for subcutaneous dosing (e.g., daily subcutaneous dosing for an appropriate period of time, e.g., for at least one day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks or more, e.g. lasting for months or years) of a long-acting compstatin analog containing a linear 40 kDa PEG and two fragments of a compstatin analog, whether as monotherapy or in combination with other therapy (eg, eculizumab therapy).

[00605] Example 22: Phase 1b LACA-40 Clinical Study in PNH Patients

[00606] An open-label phase 1b clinical trial of LACA-40 in untreated patients with PNH was performed to evaluate the safety, PK, PD, and preliminary efficacy of repeated doses of LACA-40. Male and female patients diagnosed with hemolytic PNH met the inclusion criteria. Subjects were required to have had a blood transfusion within the previous 12 months and had lactate dehydrogenase (LDH) levels >2 times the upper limit of normal (ULN). Prior to dosing, subjects were vaccinated against Neisseria meningittdes, Streptococcus pneumoniae, and Haemophilus influenzae type B (Hib) and subjects were started on prophylactic oral antibiotics. Doses of LACA-40 in 5% dextrose were administered by subcutaneous injection for at least 28 days and up to 84 consecutive days. (The term "untreated" in this context means patients who have not previously received treatment with a complement inhibitor). The study included two cohorts of 3 patients each. Doses of 180 mg/day for the first cohort and 270 mg/day for the second cohort were studied. Doses were administered in a volume of 1.8 ml as a single daily injection or as 2 daily injections of 0.9 ml. The primary efficacy endpoint for the study was the change in LDH levels as an indicator of intravascular hemolysis. Other relevant markers of efficacy measured included hemoglobin, PNH cCK clone distribution, total complement hemolytic activity (CH50), alternative pathway-mediated hemolytic activity (AP50), transfusion requirements, reticulocyte count, and deposition of C3 fragments on blood cells (as an indicator of the potential for extravascular hemolysis). ). Measurement of the deposition of C3 fragments was performed by flow cytometry, for example, as described above. The antibody used was cross-reactive with C3b, C3c and C3d.

[00607] Results

[00608] Cohort 1

[00609] Following the completion of cohort 1, two subjects received doses of LACA-40 180 mg for 28 days, and one subject withdrew informed consent due to a reaction after the first dose. In both subjects who received treatment for 28 days, there was a marked decrease in LDH levels, from 2078 U/L to 1082 U/L and from 1325 U/L to 709 U/L (normal 100 - 250 U/L). /l) from day 1 to day 29, respectively. None of the subjects met the criteria for continuing with Part 2 of the study.

[00610] Screening hemoglobin (Hb) levels were below 80 g/L and both subjects received a transfusion 3 weeks prior to dosing LACA-40. Hb levels in both subjects were maintained at 80 g/L and neither subject required transfusion during the dosing period. Both subjects received a transfusion within approximately 4 weeks of stopping LACA-40 treatment.

[00611] The cCC fraction of PNH type III (CD59-negative) in both subjects increased approximately threefold, increasing from 5.1% and 13.4% on day 1 to 17.4% and 37.6% on day 29, respectively.

[00612] In both subjects who completed 28 days of dosing, LACA-40 was safe and well tolerated. A third subject developed nausea, vomiting, and a rash 5-6 hours after receiving the 1st dose of LACA-40. This has been reported as a serious adverse event possibly related to LACA-40. To justify the continuation of the study, this reaction was further investigated by studying the reactivity of cells in vitro and performing a skin injection test with LACA-40 and 40 kDa PEG. The study concluded that there was no evidence of T cell activation and the skin test was negative. As a result, returning the subject to the study was considered medically safe. However, subject withdrew informed consent for personal reasons.

[00613] Cohort 2

[00614] After two subjects completed a period of treatment with 270 mg/day LACA-40, administered by sc injection, for 28 days, both subjects experienced a marked decrease in LDH levels to within 2-fold upper limit of normal and an increase in the percentage kKK APG type III. After analysis of all data, subjects in this cohort were eligible to continue daily LACA-40 if clinical benefit was observed and if continued use was requested by the investigator. Both treated subjects met the previously defined criteria for continued dosing up to 84 days. One subject left the study for personal reasons. The other subject continued dosing and continued to experience a sustained decrease in LDH at study day 57. Dosing of the next subject in cohort 2 was deferred.

[00615] In conclusion, LACA-40 administered daily was safe and well tolerated and resulted in sustained hemolysis suppression in PNH patients who did not receive eculizumab.

[00616] Additionally, the present disclosure specifically provides a specific understanding relating, in particular, to certain desirable characteristics of devices (e.g., needle gauge, hole diameter, and/or needle wall thickness, etc.) particularly suitable for delivery of certain LACA compositions, including, in particular, certain LACA-40 compositions described herein (see above).

[00617] Example 23: Preclinical studies of intravitreal LACA-40

[00618] Preclinical studies in monkeys were performed to evaluate the safety and pharmacology of LACA-40 when intravitreal injection. Intravitreally administered LACA-40 in cynomolgus macaques enters the vasculature and then enters further and/or is slowly excreted from the body. The results of toxicokinetic analyzes of LACA-40 concentrations in the vitreous and serum after repeated intravitreal injections for nine months at doses up to 24.8 mg/eye in 5% dextrose in volumes of 50 or 100 μl/eye indicated that during repeated injections slight intraocular or serum accumulation of the drug. In addition, a complete toxicological analysis that included indirect and slit lamp ophthalmic examination, spectral optical coherence tomography, electroretinography and ocular tonometry, and histopathological examination of both eyes and approximately 50 additional tissues from each monkey found no evidence of LACA-40- mediated changes at any of the studied doses.

[00619] Evaluation of the pharmacokinetic profile of a single intravitreal dose of LACA-40 (10 mg/eye in both eyes) in monkeys resulted in a vitreous half-life of approximately 3.2 days. Following intravitreal injection, serum concentrations of LACA-40 increased until day 7 post-dose and then decreased with an apparent half-life of 10.4 days.

[00620] Among other things, provided herein are regimens for administering a long-acting compstatin analogue comprising a linear 40 kDa PEG and two fragments of the compstatin analogue, in which the timing of individual dose administration is specifically chosen to provide the desired PK pattern given the period half-life of LACA-40 described herein.

[00621] Example 24: Phase lb clinical study of single incremental doses of LACA-40 in subjects with AMD

[00622] An open-label phase 1 clinical study of single incremental doses of LACA-40 in patients with wet AMD receiving anti-VEGF therapy (more specifically Lucentis®, Eylea® or Avastin®) was initiated to evaluate the safety, tolerability and PK of LACA-40. In this study, patients received a single dose of LACA-40 by intravitreal injection followed by 113 days of observation. Nine patients were originally planned to be included in the study, in three cohorts of three patients each, at doses of 5 mg, 10 mg, and 20 mg of LACA-40 in 5% dextrose in a volume of 100 microliters. After completion of enrollment of patients in all three cohorts, the third cohort was expanded from three patients to a total of 12 patients. LACA-40 was well tolerated by the original nine patients and no serious adverse events were reported.

[00623] The present disclosure provides administration regimens in which a long-acting compstatin analogue containing a linear 40 kDa PEG and two fragments of a compstatin analogue is administered by intravitreal injection. In certain embodiments, a long-acting compstatin analogue comprising a linear 40 kDa PEG and two fragments of the compstatin analogue is administered as monotherapy; in some embodiments, it is administered in combination with another therapy (eg, anti-VEGF therapy) such that the patient is simultaneously affected by both therapies.

[00624] This example specifically describes and justifies administration regimens in which a long-acting compstatin analogue containing a linear 40 kDa PEG and two fragments of a compstatin analogue is administered by intravitreal injection to subjects receiving anti-VEGF therapy. In certain embodiments, subjects treated with both anti-VEGF therapy and LACA-40 therapy received doses of the anti-VEGF agent at longer intervals than those used for otherwise comparable subjects not receiving LACA-40 therapy. Many anti-VEGF agents have been developed (for a review, for example, see Lanzetta Br J Opthamol 97:1497, 2013). For example, dosing regimens described in the literature for certain anti-VEGF agents include intravitreal injections of ranibizumab 0.5 mg or bevacizumab 1.25 mg given every 4 weeks (q4) or as needed; in some embodiments, such regimens serve as appropriate reference regimens against which the anti-VEGF combination therapy regimens described herein are evaluated.

[00625] In light of the description of the invention presented herein, including in this example, it will be apparent to those skilled in the art that certain combination therapy regimens have been proposed, for example, in which each of LACA-40 and an anti-VEGF agent is administered by an intravitreal route. ; in some embodiments, LACA-40 and the anti-VEGF agent may be co-administered in a single injection for certain (though not necessarily all) doses. In certain embodiments, fewer doses of the anti-VEGF agent are administered in a selected time period than would be administered in the absence of LACA-40 administration.

[00626] Example 25: Phase 2 Clinical Study of Single Incremental Doses of LACA-40 in Subjects with Geographic Atrophy

[00627] Conducted a randomized, single-blind, placebo-controlled clinical study of LACA-40 in patients with GA. Approximately 240 patients were included in the study. Patients in the study were diagnosed with macular HA due to age-related macular degeneration, which was confirmed 14 days prior to randomization at a central reading center using fundus autofluorescence imaging and the following criteria: total HA area as determined by image screening FAF, must be ≥2.5 mm ² and ≤17.5 mm ² (1 and 7 disc areas [DA], respectively).

[00628] Patients were randomized in a 2:2:1:1 ratio and patients received LACA-40 monthly, LACA-40 bimonthly, placebo injection monthly, or placebo injection bimonthly. Patients in the LACA-40 groups received a dose of 15 mg LACA-40 injected into the vitreous fluid in a volume of 0.1 cm ³ monthly or bimonthly for 12 months followed by six months of follow-up after the end of treatment. In cohorts receiving placebo injections, patients received a sham injection. The safety, tolerability, PK, and activity evidence of multiple intravitreal injections of LACA-40 in patients with GA in at least one eye was assessed. The primary efficacy endpoint was change in GA lesion size from baseline to month 12. The study design was designed to detect a reduction in lesion size growth of at least 30% between LACA-40 and placebo-controlled groups from baseline to month 12 The primary safety end point was the number and severity of local and systemic adverse events that occurred after the start of treatment.

[00629] Example 26: Phase 2 Clinical Study of Single Incremental Doses of LACA-40 in Subjects with Intermediate AMD

[00630] Conducted a randomized, single-blind, placebo-controlled clinical study of LACA-40 in patients with intermediate AMD. Patients in the study were diagnosed with intermediate age-related macular degeneration. The aim of the study was to evaluate whether LACA-40 was able to prevent progression from intermediate AMD to HA or wet AMD.

[00631] Patients were randomized in a 2:2:1:1 ratio and patients received LACA-40 monthly, LACA-40 bimonthly, placebo injection monthly, or placebo injection bimonthly. Patients in the LACA-40 groups received a dose of 15 mg LACA-40 injected into the vitreous fluid in a volume of 0.1 cm ³ monthly or every two months for at least 12 months followed by six months of follow-up after the end of treatment . In cohorts receiving placebo injections, patients received a sham injection. The safety, tolerability, and evidence of activity of multiple intravitreal injections of LACA-40 were assessed in patients with intermediate AMD in at least one eye. Efficacy endpoints included the rate of progression from intermittent AMD to LA and/or wet AMD, with a reduction in frequency in patients treated with LACA-40 compared to patients treated with placebo injections, providing evidence of efficacy, and change in amount and/or volume of high-risk drusen; and total number and/or volume of drusen. The greater reduction in the number, volume, and/or growth rate of drusen in patients treated with LACA-40 compared with patients who received placebo injections provides evidence of efficacy. The primary safety end point was the number and severity of local and systemic adverse events that occurred after the start of treatment.

[00632] Example 27: Preclinical studies of inhaled CA28

[00633] Single dose or repeat dose inhalation studies were performed in dogs and cynomolgus monkeys. In all animals at the highest inhalation doses studied, there were no drug-related events. These doses were 25 mg/kg/d in a 7 day repeated dose study in dogs, 80 mg/kg in a single dose study in monkeys, and 30 mg/kg/day in a 14 day repeated dose study in monkeys.

[00634] The Ascaris suum skin test was used to investigate the in vivo pharmacological effect of CA28 in cynomolgus monkeys compared to corticosteroids. Doses of nebulized CA28 at 20 mg/day or 15 mg/kg/d (for 14 consecutive days) had a pharmacological effect on the control of inflammatory cytokine levels in the lungs (as measured in bronchoalveolar lavage fluid) after allergen challenge as during a treatment period of 14 days, and within 28 days after stopping treatment after washing out the drug.

[00635] Example 28: Phase 1 Clinical Study of Single and Multiple Incremental Doses of Inhaled CA28 in Healthy Subjects

[00636] An open-label, randomized, placebo-controlled, phase 1 clinical trial of single and multiple escalating daily doses of a nebulized formulation of compstatin analog (CA28) was conducted to evaluate the safety, tolerability, and PK of single and multiple inhaled doses of the drug in healthy volunteers. A solution of CA28 in (2% glycerol) or placebo (in 2% glycerol) was administered using a PARI LC Sprint® jet nebulizer with a PARI TurboBOY® S airflow interrupter. four entities each. These subjects were administered a single dose of CA28 at doses ranging from 20 mg to 350 mg (20, 60, 150 or 350 mg) and the subjects were observed for 14 days post-treatment. In this part of the study, CA-28 was well tolerated and no serious adverse events were reported.

[00637] Four subjects were included in the first cohort of the multiple escalating dose portion of the study. These subjects received treatment with a dose of 60 mg/day of the drug for 14 consecutive days. However, after nine days of treatment at a dose of 60 mg/day, one subject developed signs and symptoms suggestive of a potential bacterial infection, which was considered possibly related to the pharmacology of the drug. The study was suspended and then restarted in subjects receiving 30 mg/day. Another subject developed signs and symptoms suggestive of a potential bacterial infection, which was considered possibly related to drug pharmacology, after 10 days of treatment at a dose of 30 mg/day. Then the study was terminated.

[00638] Both subjects responded within hours to first-line antibiotic treatment, indicative of bacterial pathogenesis. In this study, subjects were vaccinated against Neisseria meningitides and closely monitored for signs of infections. Although all bacterial cultures were negative, it is believed that Haemophilus influenzas or Streptococcus pneumonias may have been involved in the observed febrile episodes, since individuals suffering from C3 deficiency are known to be at increased risk of infection with Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae. Vaccines against these three pathogens are available and it is thought that the risk of infection can be eliminated by vaccination, possibly supplemented with prophylactic antibiotics (eg penicillin V).

[00639] Example 29: Pharmacokinetic (PK) model for LACA-40

[00640] LACA-40 is a potent and selective C3 complement inhibitor. Its high binding affinity affects the distribution of LACA-40 and its target, complement C3, as demonstrated by the dose-dependent increase in complement C3 that was observed in the single incremental dose and multiple incremental dose studies in healthy subjects described above (Example 20). The family of pharmacokinetic (PK) models that describe this relationship between a drug and its target is known as Target Mediated Drug Disposition (TMDD) (Levy, G: Pharmacologic target-mediated drug disposition. Clin. Pharmacol. Ther (1994) 56: 248-252). A schematic diagram of this model is shown in Fig. 15. Data from the completed single incremental dose and multiple incremental dose studies of LACA-40 in healthy subjects described in Example 20 were used to develop a TMDD model for LACA-40. The FA of LACA-40, together with the effect of C3 complement binding, has been well described using TMDD (QSS approximation, Mager DE, JuskoWJ: General pharmacokinetic model for drugs exhibiting target mediated drug disposition. J. Pharmacokinet. Pharmacodyn. (2001) 28: 507-532 ). In FIG. 16 shows the observed and predicted serum concentrations of LACA-40 during the nominal time after administration, divided by dosing regimen, for the single dose (s.d.) and multiple dose study over 28 days (ppm, once per day) in healthy subjects. The figure demonstrates that the TMDD model provides good model fit across time, dose, and schedule.

[00641] The TMDD model was used to predict serum LACA-40 residual concentrations for daily dosing of 180 mg, 270 mg, or 360 mg (FIG. 17) and to predict serum LACA-40 concentration for several different dose levels when administered three times a week, twice a week or weekly. Model predictions are shown in Fig. 18 and 19 (for administration three times a week and twice a week, respectively) and in FIG. 20 and 21 (for weekly administration). All predicted values presented are based on model assumptions from TMDD; they are generally referred to as "usual value" and can be considered equivalent to the median.

[00642] Example 30: Pharmacokinetic and pharmacodynamic studies of various treatment regimens

[00643] A phase I, double-blind, randomized, clinical trial was initiated to evaluate the safety, tolerability, PK, and PD of various LACA-40 regimens in healthy volunteers. This study included three or more cohorts dosed as described in the following tables 13-15.

[00644]

[00645] Prior to the first administration, subjects were vaccinated against Neisseria meningitides, Streptococcus pneumoniae, and Haemophilus influenzae type B (Hib).

[00646] Participants in Cohort 1 and Cohort 2 were randomized according to a 4:1 randomization ratio such that four participants received LACA-40 and one participant received placebo. Cohort 3 included two signal cohorts; randomized 1:1 with one participant receiving LACA-40 and another participant receiving placebo; after which four participants were randomized in a 3:1 ratio with three participants receiving LACA-40 and 1 participant receiving placebo. The number of participants and randomization ratio for cohort 4 (which is an optional cohort) is to be determined.

[00647]

[00648] For cohort 1, a 36 mg/mL LACA-40 solution was used. For cohorts 2 and 3, a 65 mg/mL LACA-40 solution was used. In each case, the compound was administered in sorbitol acetate buffer at pH 5.0. Subjects in Cohort 1 were administered LACA-40 or placebo daily for 28 days using a Neria™ infusion set with a 29G needle. Subjects in cohort 2 were administered LACA-40 or placebo using a Precision Flow rate tubing RM-F2400 (LACA-40) and RM-F60 (placebo) kit and one High Flo 24G 9 mm needle. Subjects in cohort 3 were administered LACA-40 or placebo using a Precision Flow rate tubing RM-F2400 (LACA-40) and RM-F60 (placebo) kit and a single needle or High Flo 24G 9 mm two-needle kit. For cohort 3, 40 ml was delivered using two syringes, with the second syringe attached to the same infusion tubing immediately after the first syringe was empty. The schedule of injections and planned sites of administration for cohorts 1, 2 and 3 are presented in the table below, where "quadrant" refers to the abdominal quadrant.

[00649]

[00650] All drugs were administered at the study site and subjects remained in the clinic until Day 35. Safety was assessed during clinical monitoring, including evaluation of the infusion site. Serial blood serum samples were taken to determine the concentrations of LACA-40.

[00651] Results

[00652] When the introduction of the first three cohorts was completed, the analysis showed that LACA-40 was safe and, in general, well tolerated.

[00653] PK data from cohorts 1 and 2 were analyzed and compared with the predictions of the TMDD model described above. Fig. 22 and 23 are graphs showing predictions. Individual and mean pooled observed PK data for cohorts 1 and 2 are shown in FIG. 24. The observed data for cohort 1 (360 mg/d) PK were consistent with the predictions of the TMDD model (FIG. 25). The data observed for cohort 1 FC at late time points were not significantly lower; than predicted by the model; but did not go beyond the expected variability/tolerance (Fig. 25). The PK data observed for cohort 2 (1300 mg twice weekly) were highly consistent with the predictions of the TMDD model (FIG. 26). At later time points, there was some evidence of a marginally higher observed PK, but the results were within expected variability/tolerance (eg, within-subject vs. between-study variations, number of subjects, etc.). The mean observed data for cohorts 1 and 2 are shown in FIG. 27 without individual data to provide better visualization and further demonstrate that the observed PK data are consistent with predictions. In FIG. Table 28 shows the observed total PK data from cohorts 1 and 2 superimposed on the prediction for cohort 3. The following table 16 shows the observed and predicted Cmax and area under the curve (AUC) over 35 days (816 hours) for cohorts 1 and 2 and predicted values for cohort 3. When compared with the results of a previous study with the introduction of 270 mg c.d. s.c. (Cmax: 670 µg/mL) the results demonstrate that in this PK study, exposure in the 360 mg q.d. was less than expected, but did not go beyond the expected variability/tolerance (for example, variations for a given subject and variations between studies, number of subjects, etc.). Observed Cmax for 1300 mg 2 r./n. was higher than predicted, which was due to the presence of one subject with a Cmax of 835 µg/ml.

[00654]

[00655] The data obtained in the described example further confirms that pharmacologically relevant doses of LACA-40 administered by subcutaneous infusion are safe and well tolerated, and also demonstrates that LACA-40 is compatible with administration by subcutaneous infusion using various pumping systems. devices and at less frequent administration than the daily administration used in previous studies involving subcutaneous administration. The data further support the ability of the TMDD pharmacokinetic model described above to accurately predict the significant PK parameters of LACA-40 for different dosing schedules, eg dosing regimens with more than a day interval between dosing (eg twice a week).

[00656] Those skilled in the art are aware of many equivalents of the specific embodiments of the present invention described in this application, or such equivalents can be identified using no more than routine experimentation. It is assumed that the scope of the present invention is not limited to the above description and is reflected in the attached claims. It should be understood that the present invention is in no way dependent on the specific results obtained in any particular example or in any particular embodiment. The use of singular nouns and the term "specified/s" may mean "one/one or more than one/one", unless otherwise indicated or obvious from the context. Claims or descriptions that include the conjunction "or" between one or more members of the group are intended to be satisfied if one or more are present, use(s), or are otherwise associated with a particular product or process. than one or all members of the specified troupe, unless otherwise indicated or evident from the context. The present invention includes embodiments in which exactly one member of the specified group is present, used, or otherwise associated with a particular product or process. For example, without limitation, it should be understood that in cases where the claims or description indicates that the residue at a particular position can be selected from a particular group of amino acids or amino acid analogs, the present invention includes individual implementations in which the residue at the specified position is any of the listed amino acids or analogs of amino acids. The present invention also includes embodiments in which more than one or all of the members of the specified group are(s) present, used or otherwise associated with a particular product or process. In addition, it should be understood that the present invention covers all variants, combinations and permutations in which one or more of the limitations, elements, conditions, descriptive terms, and the like, from one or more of the following claims or the above description is introduced into another claim in the formula. For example, any claim that is dependent on another claim may be modified to include one or more of the elements, limitations, conditions, or descriptive terms present in any other claim that is dependent on the same base claim. In addition, in cases where a composition is mentioned in the claims, it should be understood that the scope of the present invention includes methods of administering said composition, corresponding to any of the methods described in this application, and methods of using said composition for any of the purposes described in this application. ; Included within the scope of the present invention are methods of preparing a composition according to any of the methods of preparation described in this application, unless otherwise indicated, or unless it is obvious to a person skilled in the art that a contradiction or inconsistency will arise. Methods for treating a subject may include the step of providing a subject in need of such treatment (e.g., a subject who has had a disease, or a subject who has an increased risk of a disease), diagnosing a disease in a subject, and/or selecting a subject for treatment with a cellular reactive analog. compstatin. Where elements are presented as lists, it is to be understood that each subgroup of said elements is also disclosed, and that any element(s) may be removed from said group. For brevity, only some of these implementation options are specifically presented in this application, however, the present invention includes all such implementation options. It should also be understood that, in general, when the present invention or aspects of the present invention are said to include(s) specific elements, properties, and the like, certain embodiments of the present invention or aspects of the present invention consist, or consist essentially, from such elements, properties, etc. Descriptions of various diseases, disorders and conditions in various sections of this application are provided for convenience and are not intended to limit the present invention.

Limits are included when specifying ranges. In addition, it should be understood that, unless otherwise indicated or obvious from the context and for a person skilled in the art, values expressed in terms of ranges, according to various embodiments of the present invention, can mean any specific value or subrange within the specified ranges, up to tenths of units of the lower limit of the specified range, unless the context clearly implies otherwise. Any particular implementation, aspect, element, feature, or the like. of the present invention may be explicitly excluded from the claims, even if such an exception is not explicitly stated in this application. For example, any compstatin analog, functional group, linking moiety, clearance-reducing moiety, disease, or indication may be explicitly excluded.

--->

SEQUENCE LIST

<110> APELLIS PHARMACEUTICALS, INC.

<120> ADMINISTRATION REGIMENS AND RELATED COMPOSITIONS AND METHODS

<130> 2008575-0280

<140> PCT/US2018/026753

<141> 2018-04-09

<150> 62/485.343

<151> 2017-04-13

<150> 62/483.295

<151> 2017-04-07

<160> 74

<170> PatentIn version 3.5

<210> 1

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<400> 1

Ile Cys Val Val Gln Asp Trp Gly His His Arg Cys Thr

1 5 10

<210> 2

<211> 42

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (1)..(19)

<223> Any independently selected amino acid or analogue

amino acids

<220>

<221> additional_characteristic

<222> (1)..(19)

<223> This area can contain from 2 to 19 residues, and

some leftovers may be missing

<220>

<221> MOD_RES

<222> (22)..(22)

<223> Trp or equivalent Trp

<220>

<221> MOD_RES

<222> (24)..(42)

<223> Any independently selected amino acid or analogue

amino acids

<220>

<221> additional_characteristic

<222> (24)..(42)

<223> This area can contain from 2 to 19 residues, and

some leftovers may be missing

<220>

<223> For a detailed description of substitutions and preferred embodiments, see

in the filed description of the invention

<400> 2

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

1 5 10 15

Xaa Xaa Xaa Gln Asp Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

35 40

<210> 3

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(1)

<223> Trp, Trp analog, or other amino acid or amino acid analog,

containing at least one aromatic ring

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Trp, Trp analog, or other amino acid or amino acid analog,

containing at least one aromatic ring

<220>

<223> For a detailed description of substitutions and preferred embodiments, see

in the filed description of the invention

<400> 3

Xaa Gln Asp Xaa Gly

fifteen

<210> 4

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(1)

<223> Trp, Trp analog, or other amino acid or amino acid analog,

containing at least one aromatic side chain

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Trp, Trp analog, or other amino acid or amino acid analog,

containing at least one aromatic side chain

<220>

<221> MOD_RES

<222>(6)..(6)

<223> His, Ala, Ala analog, Phe, Trp or Trp analog

<220>

<223> For a detailed description of substitutions and preferred embodiments, see

in the filed description of the invention

<400> 4

Xaa Gln Asp Xaa Gly Xaa

fifteen

<210> 5

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(3)

<223> Any independently selected amino acid or analogue

amino acids

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Trp or equivalent Trp

<220>

<221> MOD_RES

<222>(7)..(7)

<223> Trp or equivalent Trp

<220>

<221> MOD_RES

<222> (9)..(13)

<223> Any independently selected amino acid or analogue

amino acids

<220>

<223> See the filed specification for details.

substitutions and preferred embodiments

<400> 5

Xaa Xaa Xaa Xaa Gln Asp Xaa Gly Xaa Xaa Xaa Xaa Xaa

1 5 10

<210> 6

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(2)

<223> Ile, Val, Leu, B1-Ile, B1-Val, B1-Leu or dipeptide,

containing Gly-Ile or B1-Gly-Ile, where B1 is

first blocking fragment

<220>

<221> additional_characteristic

<222> (1)..(2)

<223> This area can contain from 1 to 2 residues, with some

provisions may not be

<220>

<221> additional_characteristic

<222> (3)..(13)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (5)..(5)

<223> Independently selected from Trp or Trp analogs

<220>

<221> MOD_RES

<222>(8)..(8)

<223> Independently selected from Trp or Trp analogs

<220>

<221> MOD_RES

<222> (10)..(10)

<223> His, Ala or equivalent Ala, Phe, Trp or equivalent

trp

<220>

<221> MOD_RES

<222> (14)..(16)

<223> L-Thr, D-Thr, Ile, Val, Gly, a dipeptide which is selected from Thr-Ala

or Thr-Asn, or a tripeptide containing Thr-Ala-Asn, where

c-terminal -OH any of L-Thr, D-Thr, Ile, Val, Gly,

Ala or Asn is optionally substituted by the second blocking fragment

<220>

<221> additional_characteristic

<222> (14)..(16)

<223> This area can contain from 1 to 3 residues, with some

provisions may not be

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 6

Xaa Xaa Cys Val Xaa Gln Asp Xaa Gly Xaa His Arg Cys Xaa Xaa Xaa

1 5 10 15

<210> 7

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(2)

<223> Ile, Val, Leu, Ac-Ile, Ac-Val, Ac-Leu or dipeptide

containing Gly-Ile or Ac-Gly-Ile

<220>

<221> additional_characteristic

<222> (1)..(2)

<223> This area can contain from 1 to 2 residues, with some

provisions may not be

<220>

<221> additional_characteristic

<222> (3)..(13)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (5)..(5)

<223> Independently selected from Trp or Trp analogs

<220>

<221> MOD_RES

<222>(8)..(8)

<223> Independently selected from Trp or Trp analogs

<220>

<221> MOD_RES

<222> (10)..(10)

<223> His, Ala or equivalent Ala, Phe, Trp or equivalent

trp

<220>

<221> MOD_RES

<222> (14)..(16)

<223> L-Thr, D-Thr, Ile, Val, Gly, a dipeptide which is selected from Thr-Ala

or Thr-Asn, or a tripeptide containing Thr-Ala-Asn, where

c-terminal -OH any of L-Thr, D-Thr, Ile, Val, Gly, Ala, or Asn

optionally substituted with -NH2

<220>

<221> additional_characteristic

<222> (14)..(16)

<223> This area can contain from 1 to 3 residues, with some

provisions may not be

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 7

Xaa Xaa Cys Val Xaa Gln Asp Xaa Gly Xaa His Arg Cys Xaa Xaa Xaa

1 5 10 15

<210> 8

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal H

<220>

<223> C-terminal CONH2

<400> 8

Ile Cys Val Val Gln Asp Trp Gly His His Arg Cys Thr

1 5 10

<210> 9

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<223> C-terminal CONH2

<400> 9

Ile Cys Val Val Gln Asp Trp Gly His His Arg Cys Thr

1 5 10

<210> 10

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<223> C-terminal CONH2

<400> 10

Ile Cys Val Tyr Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 11

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<223> C-terminal COOH

<400> 11

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 12

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<223> C-terminal CONH2

<400> 12

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 13

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (13)..(13)

<223> d-Thr

<220>

<223> C-terminal COOH

<400> 13

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 14

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 2-Nal

<220>

<223> C-terminal CONH2

<400> 14

Ile Cys Val Ala Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 15

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 2-Nal

<220>

<223> C-terminal COOH

<400> 15

Ile Cys Val Ala Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 16

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 1-Nal

<220>

<223> C-terminal COOH

<400> 16

Ile Cys Val Ala Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 17

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 2-Needle

<220>

<223> C-terminal CONH2

<400> 17

Ile Cys Val Gly Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 18

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 2-Needle

<220>

<223> C-terminal COOH

<400> 18

Ile Cys Val Gly Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 19

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Dht

<220>

<223> C-terminal COOH

<400> 19

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 20

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Bpa

<220>

<223> C-terminal COOH

<400> 20

Ile Cys Val Phe Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 21

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Bpa

<220>

<223> C-terminal CONH2

<400> 21

Ile Cys Val Phe Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 22

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Bta

<220>

<223> C-terminal COOH

<400> 22

Ile Cys Val Xaa Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 23

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Bta

<220>

<223> C-terminal CONH2

<400> 23

Ile Cys Val Xaa Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 24

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222>(9)..(9)

<223> 2-Abu

<220>

<223> C-terminal CONH2

<400> 24

Ile Cys Val Trp Gln Asp Trp Gly Xaa His Arg Cys Thr

1 5 10

<210> 25

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal H

<220>

<223> C-terminal COOH

<400> 25

Gly Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Ala Asn

1 5 10 15

<210> 26

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 5f-Trp

<220>

<223> C-terminal CONH2

<400> 26

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 27

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 5-methyl-Trp

<220>

<223> C-terminal CONH2

<400> 27

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 28

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 1-methyl-Trp

<220>

<223> C-terminal CONH2

<400> 28

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 29

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222>(7)..(7)

<223> 5f-Trp

<220>

<223> C-terminal CONH2

<400> 29

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 30

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 5f-Trp

<220>

<221> MOD_RES

<222>(7)..(7)

<223> 5f-Trp

<220>

<223> C-terminal CONH2

<400> 30

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 31

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 5-methyl-Trp

<220>

<221> MOD_RES

<222>(7)..(7)

<223> 5f-Trp

<220>

<223> C-terminal CONH2

<400> 31

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 32

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 1-methyl-Trp

<220>

<221> MOD_RES

<222>(7)..(7)

<223> 5f-Trp

<220>

<223> C-terminal CONH2

<400> 32

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 33

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal H

<220>

<221> MOD_RES

<222> (5)..(5)

<223> 6f-Trp

<220>

<221> MOD_RES

<222>(8)..(8)

<223> 6f-Trp

<220>

<223> C-terminal COOH

<400> 33

Gly Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Asn

1 5 10 15

<210> 34

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 1-formyl-Trp

<220>

<223> C-terminal CONH2

<400> 34

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 35

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (4)..(4)

<223> 1-methoxy-Trp

<220>

<223> C-terminal CONH2

<400> 35

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 36

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal H

<220>

<221> MOD_RES

<222> (5)..(5)

<223> 5f-Trp

<220>

<221> MOD_RES

<222>(8)..(8)

<223> 5f-Trp

<220>

<223> C-terminal COOH

<400> 36

Gly Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Asn

1 5 10 15

<210> 37

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<400> 37

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 38

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)5-C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 38

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 39

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal (C=O)-(CH2)5-maleimide

<400> 39

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 40

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)2-C(=O)-NH-CH2CH2OCH2CH2O

CH2CH2C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 40

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 41

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)2-C(=O)-NH-CH2CH2OCH2CH2C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 41

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 42

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)3-C(=O)-NH-CH2CH2OCH2CH2

OCH2C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 42

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 43

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)5-C(=O)-NH-CH2CH2OCH2CH2

OCH2C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 43

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 44

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)4-C(=O)-NH-CH2CH2OCH2CH2

OCH2CH2C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 44

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 45

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)2-C(=O)-NH-CH2CH2OCH2CH2

OCH2CH2C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 45

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 46

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal maleimide-(CH2)5-C(=O)-NH-CH2CH2OCH2CH2

OCH2C(=O)

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 46

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 47

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 47

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 48

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 48

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Gly Gly Gly

1 5 10 15

Gly Gly Lys

<210> 49

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal NH2(CH2)5C(=O)

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 49

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 50

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal NH2(CH2CH2O)2CH2C(=O)

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 50

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 51

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> NH-CH2CH2OCH2CH2OCH2-C(=O)-Lys

<220>

<223> C-terminal NH2

<400> 51

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 52

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> Lys-(C(=O)-(CH2)5-Mal)

<220>

<223> C-terminal NH2

<400> 52

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 53

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (19)..(19)

<223> Lys-(C(=O)-(CH2)5-Mal)

<220>

<223> C-terminal NH2

<400> 53

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Gly Gly Gly

1 5 10 15

Gly Gly Lys

<210> 54

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Mal-(CH2)5-(C(=O)-NH(CH2)5C(=O)

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 54

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 55

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Mal-(CH2)5-(C(=O)NH(CH2CH2O)2CH2C(=O)

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<400> 55

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 56

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> NH-CH2CH2OCH2CH2OCH2-C(=O)-Lys-(C(=O)-(CH2)5-Mal)

<220>

<223> C-terminal NH2

<400> 56

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 57

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> Lys-C(=O)-CH2(OCH2CH2)2NH(C(=O)-(CH2)5-Mal)

<220>

<223> C-terminal NH2

<400> 57

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 58

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal (CH2CH2O)nC(=O)

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 58

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 59

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> NH-CH2CH2OCH2CH2OCH2-C(=O)-Lys-C(=O)-(CH2CH2O)n

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 59

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 60

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> Lys-C(=O)-(CH2CH2O)n

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 60

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 61

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (19)..(19)

<223> Lys-C(=O)-(CH2CH2O)n

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 61

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Gly Gly Gly

1 5 10 15

Gly Gly Lys

<210> 62

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (1)..(1)

<223> (CH2CH2O)nC(=O)Lys

<220>

<221> additional_characteristic

<222> (8)..(18)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (10)..(10)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 62

Lys Gly Gly Gly Gly Gly Ile Cys Val Trp Gln Asp Trp Gly Ala His

1 5 10 15

Arg Cys Thr

<210> 63

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> MOD_RES

<222> (1)..(1)

<223> (CH2CH2O)nC(=O)Lys

<220>

<221> additional_characteristic

<222> (3)..(13)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (5)..(5)

<223> (1Me)Trp

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 63

Lys Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr

1 5 10

<210> 64

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> NH-CH2CH2OCH2CH2OCH2-C(=O)-Lys-(C(=O)-(CH2CH2O)n-R)

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 64

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 65

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> NH-CH2CH2OCH2CH2OCH2-C(=O)-Lys-(C(=O)-(CH2)m-

(CH2CH2O)n-R)

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 65

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 66

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> NH-CH2CH2OCH2CH2OCH2-C(=O)-Lys-(C(=O)-(CH2)m-C(=O)-

(CH2CH2O)n-R)

<220>

<223> C-terminal NH2

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 66

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 67

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> MOD_RES

<222> (14)..(14)

<223> NH-CH2CH2OCH2CH2OCH2-C(=O)-Lys-(C(=O)-(CH2)m-C(=O)

-(CH2)j(CH2CH2O)n-R)

<220>

<223> C-terminal NH2

<400> 67

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 68

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<223> N-terminal Ac

<220>

<221> additional_characteristic

<222> (2)..(12)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (4)..(4)

<223> (1Me)Trp

<220>

<221> additional_characteristic

<222> (13)..(14)

<223> Residues at these positions are connected using the AEEAc fragment

<220>

<221> MOD_RES

<222> (14)..(14)

<223> Lys-(C(=O)-(CH2)5-Mal)

<220>

<223> C-terminal NH2

<400> 68

Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Thr Lys

1 5 10

<210> 69

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

6xHis-label

<400> 69

His His His His His His His

fifteen

<210> 70

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(1)

<223> Any amino acid

<220>

<221> MOD_RES

<222> (2)..(4)

<223> Any independently selected amino acid or analogue

amino acids

<220>

<221> MOD_RES

<222> (5)..(5)

<223> Trp or equivalent Trp

<220>

<221> MOD_RES

<222>(8)..(8)

<223> Trp or equivalent Trp

<220>

<221> MOD_RES

<222> (10)..(14)

<223> Any independently selected amino acid or analogue

amino acids

<220>

<221> MOD_RES

<222> (15)..(15)

<223> Any amino acid

<220>

<223> Detailed description of replacements and preferred embodiments

see the filed description of the invention

<400> 70

Xaa Xaa Xaa Xaa Xaa Gln Asp Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa

1 5 10 15

<210> 71

<211> 18

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(1)

<223> Any amino acid

<220>

<221> MOD_RES

<222> (2)..(3)

<223> Ile, Val, Leu, B1-Ile, B1-Val, B1-Leu or dipeptide,

containing Gly-Ile or B1-Gly-Ile, where B1 is

first blocking fragment

<220>

<221> additional_characteristic

<222> (2)..(3)

<223> This area can contain from 1 to 2 residues, with some

provisions may not be

<220>

<221> additional_characteristic

<222> (4)..(14)

<223> Disulfide bond

<220>

<221> MOD_RES

<222>(6)..(6)

<223> Independently selected from Trp or Trp analogs

<220>

<221> MOD_RES

<222>(9)..(9)

<223> Independently selected from Trp or Trp analogs

<220>

<221> MOD_RES

<222> (11)..(11)

<223> His, Ala or equivalent Ala, Phe, Trp or equivalent

trp

<220>

<221> MOD_RES

<222> (15)..(17)

<223> L-Thr, D-Thr, Ile, Val, Gly, a dipeptide which is selected from Thr-Ala

or Thr-Asn, or a tripeptide containing Thr-Ala-Asn, where

c-terminal -OH any of L-Thr, D-Thr, Ile, Val, Gly,

Ala or Asn is optionally substituted by the second blocking fragment

<220>

<221> additional_characteristic

<222> (15)..(17)

<223> This area can contain from 1 to 3 residues, with some

provisions may not be

<220>

<221> MOD_RES

<222> (18)..(18)

<223> Any amino acid

<220>

<223> For a detailed description of substitutions and preferred embodiments, see

filed description of the invention

<400> 71

Xaa Xaa Xaa Cys Val Xaa Gln Asp Xaa Gly Xaa His Arg Cys Xaa Xaa

1 5 10 15

Xaa Xaa

<210> 72

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(1)

<223> Any amino acid

<220>

<221> MOD_RES

<222> (2)..(2)

<223> Ile or Gly

<220>

<221> MOD_RES

<222> (3)..(3)

<223> Ile or Cys

<220>

<221> MOD_RES

<222> (4)..(4)

<223> Val or Cys

<220>

<221> MOD_RES

<222> (5)..(5)

<223> Val, Tyr, Trp, 2-Nal, 1-Nal, 2-Igl, Dht, Bpa, Bta,

5f-Trp, 5-methyl-Trp, 1-methyl-Trp, 1-formyl-Trp or

1-methoxy-Trp

<220>

<221> MOD_RES

<222>(6)..(6)

<223> Gln, Trp, 6f-Trp or 5f-Trp

<220>

<221> MOD_RES

<222>(7)..(7)

<223> Asp or Gln

<220>

<221> MOD_RES

<222>(8)..(8)

<223> Trp, Asp or 5f-Trp

<220>

<221> MOD_RES

<222>(9)..(9)

<223> Gly, Trp, 6f-Trp or 5f-Trp

<220>

<221> MOD_RES

<222> (10)..(10)

<223> His, Ala, 2-Abu or Gly

<220>

<221> MOD_RES

<222> (11)..(11)

<223> His or Ala

<220>

<221> MOD_RES

<222> (12)..(12)

<223> Arg or His

<220>

<221> MOD_RES

<222> (13)..(13)

<223> Cys or Arg

<220>

<221> MOD_RES

<222> (14)..(14)

<223> Thr, D-Thr or Cys

<220>

<221> MOD_RES

<222> (15)..(15)

<223> Any amino acid

<220>

<223> For a detailed description of substitutions and preferred embodiments, see

filed description of the invention

<400> 72

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

1 5 10 15

<210> 73

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(1)

<223> Any amino acid

<220>

<221> additional_characteristic

<222> (3)..(13)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (5)..(5)

<223> Trp(Me)

<220>

<221> MOD_RES

<222>(9)..(9)

<223>mGly

<220>

<221> MOD_RES

<222> (14)..(14)

<223> miles

<220>

<221> MOD_RES

<222> (15)..(15)

<223> Any amino acid

<220>

<223> For a detailed description of substitutions and preferred embodiments, see

filed description of the invention

<400> 73

Xaa Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Ile Xaa

1 5 10 15

<210> 74

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Artificial sequence description: Synthetic

peptide

<220>

<221> MOD_RES

<222> (1)..(1)

<223> Any amino acid

<220>

<221> additional_characteristic

<222> (3)..(13)

<223> Disulfide bond

<220>

<221> MOD_RES

<222> (5)..(5)

<223> Trp(Me)

<220>

<221> MOD_RES

<222>(9)..(9)

<223>mGly

<220>

<221> MOD_RES

<222> (14)..(14)

<223> miles

<220>

<221> MOD_RES

<222> (15)..(15)

<223> Any amino acid

<220>

<223> For a detailed description of substitutions and preferred embodiments, see

filed description of the invention

<400> 74

Xaa Ile Cys Val Trp Gln Asp Trp Gly Ala His Arg Cys Ile Xaa

1 5 10 15

<---

Claims (42)

1. A method of treating a subject in need of treatment for a complement-mediated disorder comprising administering a long acting compstatin analogue (LACA) to said subject subcutaneously according to a dosing regimen in which 1080 mg of LACA is administered thrice a week, once every three days, twice weekly or weekly wherein LACA comprises a clearance reducing fragment fused to two fragments of a compstatin analog, each fragment of the compstatin analog comprising a cyclic peptide comprising the amino acid sequence shown in SEQ ID NO: 28 extended with a lysine residue or a sequence containing a lysine residue at the N-terminus, C-terminus or both ends, and the specified lysine residue is separated from the cyclic part of the specified peptide by a spacer containing 8-amino-3,6-dioxaoctanoic acid (AEEAc); and said clearance-reducing fragment contains a polymer, where each end of the specified polymer is associated with one of the specified fragments of the analog compstatin carbamate, and wherein said polymer is PEG with an average molecular weight of 40 kDa. 2. The method according to claim 1, characterized in that LACA is administered weekly. 3. The method according to p. 1, characterized in that LACA is administered twice a week. 4. The method according to p. 1, characterized in that LACA is administered three times a week. 5. The method according to p. 1, characterized in that LACA is administered once every three days. 6. The method according to claim 1, characterized in that said complement-mediated disorder is hemolytic anemia, paroxysmal nocturnal hemoglobinuria (PNH), myasthenia gravis, glomerulonephritis, NZN (optic neuromyelitis), polyneuropathy, nephropathy, or vasculitis. 7. The method of claim 1, wherein said complement-mediated disorder is PNH. 8. The method according to p. 1, characterized in that LACA contains a compound having the structure:

, moreover, the specified polymer is a PEG with an average molecular weight of 40 kDa. 9. A single dosage composition of LACA for the treatment of a complement-mediated disorder in a subject in need thereof, wherein said single dosage composition is for subcutaneous administration, and wherein the amount of said single dosage composition is from 1080 mg, moreover, LACA contains a clearance-reducing fragment attached to two fragments of a compstatin analog, and each fragment of the compstatin analog contains a cyclic peptide containing the amino acid sequence shown in SEQ ID NO: 28, extended by a lysine residue or a sequence containing a lysine residue, at the N-terminus, C-terminus, or both ends, and the specified lysine residue is separated from the cyclic parts of said peptide with a spacer containing 8-amino-3,6-dioxaoctanoic acid (AEEAc); and wherein said clearance-reducing fragment contains a polymer, where each end of the specified polymer is associated with one of the specified fragments of the analog compstatin carbamate, and moreover, the specified polymer is a PEG with an average molecular weight of 40 kDa. 10. A single dosage composition according to claim 9, characterized in that said single dosage composition is intended for subcutaneous administration once every three days. 11. A unit dosage composition according to claim 9, characterized in that said unit dosage composition is for weekly subcutaneous administration. 12. A single dosage composition according to claim 9, characterized in that said single dosage composition is intended for subcutaneous administration twice a week. 13. A single dosage composition according to claim 9, characterized in that said single dosage composition is intended for subcutaneous administration three times a week. 14. A single dosage composition according to claim 9, additionally containing a pharmaceutically acceptable carrier. 15. Single dosage composition according to claim 9, characterized in that LACA contains a compound having the structure:

, moreover, the specified polymer is a PEG with an average molecular weight of 40 kDa. 16. A drug delivery device containing a single dosage composition according to claim 9. 17. The device according to claim 16, characterized in that said device is a syringe or container. 18. A method of treating a subject in need of treatment for a complement-mediated disorder, comprising subcutaneously administering a single dosage composition according to claim 9 to said subject. 19. The method according to p. 18, characterized in that the specified unit dosage composition is administered using a syringe pump. 20. The method of claim 18, wherein said unit dosage composition is administered using a wearable delivery device. 21. The method according to p. 18, characterized in that said single dosage composition is administered once every three days. 22. The method of claim. 18, characterized in that said single dosage composition is administered weekly. 23. The method according to p. 18, characterized in that the specified single dosage composition is administered twice a week. 24. The method according to p. 18, characterized in that the specified single dosage composition is administered three times a week. 25. The method according to claim 18, characterized in that said complement-mediated disorder is hemolytic anemia, paroxysmal nocturnal hemoglobinuria (PNH), myasthenia gravis, glomerulonephritis, NZN (optic neuromyelitis), polyneuropathy, nephropathy, or vasculitis. 26. The method of claim 18, wherein said complement-mediated disorder is PNH. 27. The method according to p. 18, characterized in that LACA contains a compound having the structure:

, moreover, the specified polymer is a PEG with an average molecular weight of 40 kDa.

Images