Compositions and methods for treating eye diseases

Abstract

The present disclosure generally relates to compositions and methods for preventing, reducing the risk of developing, or treating eye diseases (e.g., glaucoma or age-related macular degeneration). Age-related macular degeneration can be geographic atrophy.

Description

[Technical Field]

[0001] Related applications This patent application claims priority to U.S. Provisional Patent Application No. 63 / 121,629, filed on 4 December 2020, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Glaucoma and age-related macular degeneration (AMD) are leading causes of irreversible blindness worldwide. Geographic atrophy (GA), also known as atrophic AMD or advanced atrophic AMD, is an advanced form of AMD. The global prevalence of glaucoma in the population aged 40–80 is 3.54% (95% CrI, 2.09–5.82). Prevalence is projected to increase to 112 million worldwide and 4.7 million in North America by 2040. Currently approved treatments for glaucoma are limited to lowering intraocular pressure (IOP). Surgery, laser treatment, or IOP-lowering agents are all commonly used. However, even with good IOP management through medication or surgery, many glaucoma patients continue to experience progressive vision loss. Furthermore, there are no approved treatments or therapies for GA. Therefore, in the management of IOP, there is a significant unmet need for neuroprotective measures for patients whose treatment is insufficient to prevent ganglion cell, axon, or synaptic loss that leads to progressive and irreversible visual impairment. Thus, there is a need for novel therapies to prevent, reduce the risk of, and treat eye diseases (e.g., glaucoma and AMD such as geographic atrophy). [Overview of the Initiative]

[0003] This disclosure generally relates to compositions and methods for preventing, reducing the risk of developing, or treating eye diseases in human patients (e.g., glaucoma or age-related macular degeneration such as AMD, including geographic atrophy). Such methods include administering to a patient, via intravitreal injection, a composition comprising about 1 mg to about 10 mg of anti-C1q antibody, wherein the antibody comprises a light chain variable domain comprising HVR-L1 having the amino acid sequence of SEQ ID NO: 5, HVR-L2 having the amino acid sequence of SEQ ID NO: 6, and HVR-L3 having the amino acid sequence of SEQ ID NO: 7, and a heavy chain variable domain comprising HVR-H1 having the amino acid sequence of SEQ ID NO: 9, HVR-H2 having the amino acid sequence of SEQ ID NO: 10, and HVR-H3 having the amino acid sequence of SEQ ID NO: 11. In some embodiments, the antibody includes a light chain variable domain containing an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs. 4 and 35-38, wherein the light chain variable domain includes HVR-L1 having the amino acid sequence of SEQ ID NOs. 5, HVR-L2 having the amino acid sequence of SEQ ID NOs. 6, and HVR-L3 having the amino acid sequence of SEQ ID NOs. 7. In some embodiments, the light chain variable domain contains an amino acid sequence selected from SEQ ID NOs. 4 and 35-38. In some embodiments, the antibody includes a heavy chain variable domain containing an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs. 8 and 31-34, wherein the heavy chain variable domain includes HVR-H1 having the amino acid sequence of SEQ ID NOs. 9, HVR-H2 having the amino acid sequence of SEQ ID NOs. 10, and HVR-H3 having the amino acid sequence of SEQ ID NOs. 11. In some embodiments, the heavy chain variable domain contains an amino acid sequence selected from SEQ ID NOs. 8 and 31-34.In some embodiments, the antibody comprises a light chain variable domain having an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs. 4 and 35-38, and including HVR-L1 having the amino acid sequence of SEQ ID NOs. 5, HVR-L2 having the amino acid sequence of SEQ ID NOs. 6, and HVR-L3 having the amino acid sequence of SEQ ID NOs. 7; and a heavy chain variable domain having an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs. 8 and 31-34, and including HVR-H1 having the amino acid sequence of SEQ ID NOs. 9, HVR-H2 having the amino acid sequence of SEQ ID NOs. 11, and HVR-H3 having the amino acid sequence of SEQ ID NOs. In some embodiments, the antibody comprises a light chain variable domain having an amino acid sequence selected from SEQ ID NOs. 4 and 35-38, and a heavy chain variable domain having an amino acid sequence selected from SEQ ID NOs. 8 and 31-34. The antibody may be a monoclonal antibody, a humanized antibody, a human antibody, a chimeric antibody, an antibody fragment, or an antibody derivative thereof. The antibody fragment may be a Fab fragment, a Fab' fragment, an F(ab')2 fragment, an Fv fragment, a diabody, or a single-chain antibody molecule. In some embodiments, the Fab fragment includes the heavy chain Fab fragment of SEQ ID NO: 39 and the light chain Fab fragment of SEQ ID NO: 40.

[0004] In some embodiments, the antibody is administered once a week, once every two weeks, once every three weeks, once a month, once every four weeks, once every six weeks, once every eight weeks, once every two months, once every ten weeks, once every twelve weeks, once every three months, or once every four months. In some embodiments, the antibody is administered for at least six months, at least seven months, at least eight months, at least nine months, at least ten months, at least eleven months, or at least twelve months.

[0005] In some embodiments, the administered composition contains about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, or about 10 mg of anti-C1q antibody. The administered composition may contain about 1 mg to about 5 mg of anti-C1q antibody. The administered composition may contain about 1 mg to about 2.5 mg, about 2.5 mg to about 5 mg, about 5 mg to about 7.5 mg, or about 7.5 mg to about 10 mg of anti-C1q antibody. The administered composition may contain about 5 mg of anti-C1q antibody. The administered composition may contain about 10 mg of anti-C1q antibody.

[0006] In some embodiments, the eye disease is glaucoma or age-related macular degeneration such as geographic atrophy. [Brief explanation of the drawing]

[0007] [Figure 1] The human C1q binding assay is described below. Human C1q was bound to Mab2-Fab, FabA, and Mab2 using a one-sided ELISA. The bound antibody or Fab molecule was detected using an enzyme-labeled anti-human Fc or anti-human kappa antibody, followed by the enzyme substrate. The antibodies showed comparable binding affinity to human C1q. The EC50 values ​​for Mab2-Fab, FabA, and Mab2 were 4.4, 2.5, and 4.9 ng / mL (in the range of 34–95 pM), respectively. [Figure 2]This study demonstrates that FabA inhibits the classical pathway but not the lectin pathway or the complement accessory pathway. FabA and Mab2 were evaluated for their ability to inhibit the classical pathway, lectin pathway, and complement accessory pathway using an ELISA-based assay kit from Eurodiagnostica (Weislab®). Wells were coated with specific activators for the classical pathway (IgM), lectin pathway (mannan), or accessory pathway (lipopolysaccharide), and activation of all pathways was assessed using a C5b-9 terminal complex detection antibody. A C5 inhibitory antibody was used as a positive control. FabA and Mab2 selectively blocked the classical pathway with an IC50 of 0.3 μg / mL, while anti-C5 inhibited all three pathways. [Figure 3] This study demonstrates the inhibition of hemolysis of IgM-coated red blood cell blasts (RBCs) in human serum. Sheep RBCs, pre-sensitized with a surface-reactive polyclonal IgM antibody, were co-cultured with human serum (diluted to 100x) at 37°C for 20-30 minutes. RBC hemolysis was quantified by measuring hemoglobin release and expressed as the percentage of hemolysis caused by the absence of treatment. [Figure 4]This study shows a reduction in the number of damaged axons in the optic nerve of eyes treated with Mab1-Fab, Mab1, or Mab2. In one eye of each animal, on day 1, 1 μl of 6 μm polystyrene beads, 1 μl of 10 μm polystyrene beads (Polybead Microspheres; Polysciences, Inc., Warrington, PA, USA), and 1 μl of viscoelastic solution (10 mg / mL sodium hyaluronate; Advanced Medical Optics Inc., USA) were injected into the anterior chamber to induce an increase in IOP. The contralateral eye was left untreated to serve as a control. Antibodies Mab2, Mab1, and Mab1-Fab (Fab derived from enzymatic digestion of Mab1), versus saline, were administered to eyes with intravitreous microbead injections one day before and one week after microbead injection (days 0 and 7, 2 μL of 10 mg / mL antibody saline versus saline alone for each injection). Two weeks after injury, optic nerves were collected from animals (perfused with saline and 4% paraformaldehyde), post-fixed with 4% paraformaldehyde and 1% osmium, dehydrated at increasing alcohol concentrations, and placed in 1% uranyl acetate / ethanol. The nerves were embedded in epoxy resin, and semi-thin sections (1 μm) were excised. The total number of degenerated axons was estimated using StereoInvestigator software (MicroBrightfield, Inc., VT, USA). Scale bar = 20 μm. Both Mab1-Fab and Mab2 significantly reduced the formation of damaged axons in the optic nerve, while antibody Mab1 showed a similar trend. [Figure 5A] This study demonstrates the protection of photoreceptor nerve loss and retinal function using Mab1 antibodies in a mouse photoinjury model. The report describes a 7-day photoinjury model in mice, followed by intravitreal administration (IVT) of Mab1 antibodies, and evaluation of retinal function and histology on day 14. On day 7, 1 μL of 7.5 mg / mL of Mab1 or an isotype control antibody was administered to mice via IVT. [Figure 5B]In a mouse photoinjury model, the use of Mab1 antibody demonstrates protection against photoreceptor neuron loss and retinal function. Mab1 treatment resulted in a significant reduction in Tunel-positive photoreceptor cells in the outer granular layer of the retina compared to isotype controls. [Figure 5C] In a mouse photoinjury model, we demonstrate protection of photoreceptor nerve loss and retinal function using the Mab1 antibody. Mab1 treatment resulted in an increase in the number of photoreceptor rows in the outer granular layer compared to isotype controls. [Figure 5D] In a mouse photoinjury model, the use of Mab1 antibody demonstrates protection against photoreceptor nerve loss and retinal function. Mab1 antibody treatment resulted in a significant increase in A and B waves on the electroretinogram at day 14 compared to isotype control antibody treatment. [Figure 6] This shows the free C1q in aqueous humor after a single IVT injection. [Modes for carrying out the invention]

[0008] General Overview This disclosure generally pertains to compositions and methods for preventing, reducing the risk of developing, or treating eye diseases, such as glaucoma or age-related macular degeneration, including AMD with geographic atrophy.

[0009] Disclosed herein are recombinant humanized immunoglobulin G (IgG1) antigen-binding fragments (Fabs) that inhibit the classical complement cascade without affecting the lectin pathway or complement accessory pathway. Anti-C1q Fabs (e.g., FabA, i.e., anti-C1q Fab containing the heavy chain Fab fragment of SEQ ID NO: 39 and the light chain Fab fragment of SEQ ID NO: 40) are being developed as reagents for intravitreous (IVT) administration for the treatment of ophthalmic diseases, such as glaucoma and AMDs such as geographic atrophy (GA). A hypervariable region derived from mouse antibody M1 (Mab1 antibody containing the heavy chain variable domain of SEQ ID NO: 3 and the light chain variable domain of SEQ ID NO: 7) is described as a human IgG1 Fab fragment construct (FabA). A full-length human IgG4 antibody (Mab2, i.e., antibody containing the heavy chain variable domain of SEQ ID NO: 8 and the light chain variable domain of SEQ ID NO: 4) containing a hypervariable region derived from Mab1 is also described. Mab1 and Mab2, as well as their Fabs (Mab1-Fab and Mab2-Fab), were used as surrogate molecules for FabA in pharmacological studies. FabA is a monovalent Fab construct lacking Fc heavy chain constant domains 2 and 3 (CH2 and CH3), and therefore cannot bind to C1q via interaction with the Fc domain. Furthermore, because FabA possesses only a single antigen-binding arm, it does not exhibit agonist activity against C1q across a wide range of FabA concentrations.

[0010] The complement cascade is a crucial component of innate immunity and can be activated via three distinct pathways: the classical pathway, the lectin pathway, and the secondary pathway. All three pathways lead to the activation of complement component C3, which ultimately results in the recruitment of immune cells, inflammation, cell membrane lysis via the membrane invasion complex, and cell death.

[0011] C1q, a molecule that initiates the classical complement cascade, is thought to be associated with the initiation and progression of neurodegenerative diseases such as glaucoma and AMDs such as GA. Inhibiting C1q may delay nerve and synaptic damage by blocking the initiation of the classical complement cascade, directly reducing damage to nerve cell membranes, and reducing inflammation resulting from complement activation.

[0012] Mab2-Fab and / or FabA exhibit high affinity binding to human C1q, as measured by Biacore (<10 pM) and enzyme-linked immunosorbent assay (ELISA) (40–50 pM, Figure 1). Mab1 binds to the isolated globular head domain of C1q but not to the collagenous tail of C1q (determined by ELISA). Consistent with this finding, Mab1 inhibits substrate interactions mediated by the globular head domain of C1q (IgM, C-reactive protein [CRP], and phosphatidylserine), and FabA inhibits the functional interaction of C1q with immunoglobulin M (IgM) coated erythrocytes (RBCs) (blocking hemolysis, Figure 3). The antibody Mab1 specifically recognizes C1q and does not bind to other complement components (C3b and C5), or other C1q / tumor necrosis factor (TNF) superfamily members, including TNF, or to adiponectin, a protein that shares the highest sequence identity with C1q in its globular head domain. Consistent with these results, FabA does not inhibit the lectin complement pathway initiated by mannose-binding lectin (MBL, another C1q / TNF superfamily member), nor does it inhibit the complement activation side pathway (initiated by C3b) (Figure 2).

[0013] Glaucoma is a series of progressive eye disorders that damage the optic nerve and ultimately lead to vision loss. In primary open-angle glaucoma, the main cause of optic nerve damage is elevated intraocular pressure (IOP), which leads to a characteristic loss of visual acuity over time as measured by visual field tests. This vision loss is due to (1) progressive degeneration of retinal nerve cells or ganglion cells in the retinal ganglion cell (RGC) layer, (2) progressive degeneration of axons of retinal nerve cells or ganglion cells in the retinal nerve fiber layer (RNFL), and (3) a decrease in the number of nerve cell synapses, primarily in the intraretinal plexiform layer.

[0014] Currently approved treatments for glaucoma are limited to lowering intraocular pressure (IOP). Surgery, laser treatment, or IOP-lowering agents are all commonly used. However, even with good IOP management through medication or surgery, many glaucoma patients continue to experience progressive vision impairment. In the Collaborative Early Glaucoma Trial (CIGTS), glaucoma patients were randomized to receive either trabeculectomy (surgery) or topical medication as initial treatment. Throughout the trial, despite mean IOPs ranging from 17.1–18.3 mmHg in the medication group and 13.8–14.4 mmHg in the surgery group, the proportion of patients who substantially worsened (mean deviation of ≥3 dB on Humphrey visual field testing [HVF]) was 23.1% and 34.1%, respectively, at 9 years. In the Early-Onset Glaucoma Trial, patients were randomized to receive either argon laser trabeculoplasty combined with topical beta-blockers or to a group that received no treatment until progression was observed. In the group randomized to initial treatment, 45% of patients progressed at 6 years, based on HVF or optic nerve examination, despite a 25% reduction in IOP from baseline. These data demonstrate a significant unmet need for neuroprotective measures for patients whose IOP management is insufficient to prevent ganglion cell, axon, or synaptic loss that leads to progressive and irreversible visual impairment.

[0015] C1q recognizes certain pathogens, denatured autoantigens, antigen-binding antibodies, or specific molecules on the cell surface. In normal aging, C1q accumulates at synapses, but this may weaken due to age or stress on nerve cells, followed by various pathophysiological stimuli that can trigger activation of the classical complement cascade, leading to improper synaptic elimination. This abnormal inflammatory response accompanied by synaptic elimination is called complement-mediated neurodegeneration (CMND). CMND is involved in Alzheimer's disease, schizophrenia, Huntington's disease, frontotemporal dementia, spinal muscular atrophy, and glaucoma. In the presence of degenerative stress in the retina, C1q activation leads to synaptic elimination, contributing to RGC and optic nerve loss. Increased C1q and complement activation have been observed in the retinas of human glaucoma patients and have been demonstrated by proteomic analysis and histological staining. CMND has also been reported in rat, mouse, and dog models of glaucoma. In a mouse model of chronic, spontaneous glaucoma (DBA / 2J mice), retinal C1q accumulation and synaptic loss occurred early in disease progression, while C1q gene-deficient models were protected, with significantly delayed RGC loss and optic nerve degeneration. These results were replicated in another mouse model involving a rapid increase in IOP due to microbead injection into the anterior chamber. Both C1q gene deficiency and pharmacological inhibition using direct injection of complement inhibitors into the eye protected against neuronal loss and retinal degeneration. In another study, retinal degeneration following transient ischemia induced by a short-term hyperincrease in IOP was examined, and again, RGC loss and retinal thinning were mitigated in C1q knockout mice.

[0016] Age-related macular degeneration (AMD) is the leading cause of blindness worldwide. GA, also known as atrophic AMD or advanced atrophic AMD, is an advanced form of AMD that leads to progressive and irreversible loss of retinal central photoreceptor cells, retinal pigment epithelium, and choroidal capillaries, resulting in visual impairment. Currently, there are no approved treatments or therapies for GA.

[0017] Complement appears to be genetically linked to AMD, along with polymorphisms identified by six different proteins that can alter complement pathway activity. By inhibiting the interaction of the target-binding head of C1q, as performed by FabA, the activity of the classical pathway can be completely inhibited, leaving the lectin pathway and complement-activating accessory pathways untreated. The C1q / classical complement pathway mediates the elimination of undesirable synapses during development. In adults, C1q accumulates on synapses, and with age and disease, it can abnormally induce synaptic elimination, neuroinflammation, and degeneration. Inhibition of C1q is protective in numerous models of neurodegeneration.

[0018] C1q, a molecule that initiates the classical complement cascade, is thought to be associated with the onset and progression of GA. With aging, C1q accumulates on photoreceptor synapses and drusen in the outer plexiform layer during the progression of two distinct and important GA-related diseases. Drusens are extracellular accumulations of lipids and proteins from degenerating photoreceptor outer segments, and an increase in their size and area is associated with an increased risk of AMD development. Activation of both the classical pathway and the complement activation accessory pathway is evident on the photoreceptor outer segments and has been shown to cause retinal atrophy in GA. In a photodamage-induced GA mouse model, abnormal C1q / classical pathway activity is associated with photoreceptor loss. Localized C1q produced by retinal microglia / macrophages has been shown to be the culprit behind inflammasome activation and inflammation. In this model, inhibiting retinal C1q with FabA slowed retinal atrophy, reduced photoreceptor loss, increased retinal thickness, and preserved retinal function. FabA offers a novel approach to treating complement-mediated retinal degeneration, including GA.

[0019] All sequences cited herein are incorporated by reference from U.S. Patent Application No. 14 / 933,517, U.S. Patent Application No. 14 / 890,811, U.S. Patent No. 8,877,197, U.S. Patent No. 9,708,394, U.S. Patent Application No. 15 / 360,549, U.S. Patent No. 9,562,106, U.S. Patent No. 10,450,382, U.S. Patent No. 10,457,745, and International Patent Application No. PCT / US2018 / 022462 (each of which is incorporated herein by reference with respect to the antibodies and associated compositions disclosed therein).

[0020] Full-length antibodies can be prepared using recombinant DNA modification techniques. Such modified versions include, for example, those made from the variable region of a native antibody by insertion, deletion, or alteration within or into the amino acid sequence of the native antibody. Specific examples of this type include their modified variable region domains containing at least one CDR from a first antibody and optionally one or more framework amino acids and the remainder of the variable region domain from a second antibody. Antibody-encoding DNA can be prepared by deleting all but the desired portion of the full-length antibody-encoding DNA. Chimeric antibody-encoding DNA can be prepared by recombining DNA that substantially or exclusively encodes the human constant region with DNA encoding a variable region substantially or exclusively derived from the variable region sequence of a non-human mammal. Humanized antibody-encoding DNA can be prepared by recombining DNA encoding a variable region other than the constant region and complementarity-determining region (CDR) substantially or exclusively derived from the corresponding human antibody region with DNA encoding a CDR substantially or exclusively derived from a non-human mammal.

[0021] Suitable sources of antibody-encoding DNA molecules include cells such as hybridomas that express full-length antibodies. For example, antibodies can be isolated from host cells that express expression vectors encoding the antibody's heavy and / or light chains.

[0022] Antibody fragments, and / or antibody derivatives, including but not limited to Fab fragments, can also be prepared by the use of recombinant DNA modification techniques, including the manipulation and re-expression of DNA encoding the antibody variable and constant regions. Standard molecular biotechnologies may be used, if desired, to modify, add, or delete further amino acids or domains. Any changes to the variable or constant regions are still encompassed within the terms “variable” and “constant” regions as used herein. In some examples, C H To ensure that translation of one domain stops at interchain cysteine, C H PCR is used to generate antibody fragments by introducing a stop codon immediately after the codon encoding the interchain cysteine. Methods for designing suitable PCR primers are well known in the art, and antibody C H Single-domain sequences are readily available. In some embodiments, stop codons can be introduced using site-directed mutagenesis techniques.

[0023] The antibodies of this disclosure may be derived from any antibody isotype ("class"), including, for example, IgG, IgM, IgA, IgD, and IgE and their subclasses (including, for example, IgG1, IgG2, IgG3, and IgG4). In certain preferred embodiments, the heavy and light chains of the antibody are derived from IgG. The heavy and / or light chains of the antibody may be derived from mouse IgG or human IgG. In certain other preferred embodiments, the heavy and / or light chains of the antibody are derived from human IgG1. In yet another preferred embodiment, the heavy and / or light chains of the antibody are derived from human IgG4.

[0024] The antibodies of this disclosure may bind to C1q, C1r, or C1s and inhibit their biological activity. For example, (1) C1q binding to autoantibodies, (2) C1q binding to C1r, (3) C1q binding to C1s, (4) C1q binding to phosphatidylserine, (5) C1q binding to pentraxin-3, (6) C1q binding to C-reactive protein (CRP), (7) C1q binding to globular C1q receptor (gC1qR), (8) C1q binding to complement receptor 1 (CR1), (9) C1q binding to B amyloid, or (10) C1q binding to calreticulin.In other embodiments, the biological activity of C1q includes: (1) activation of the classical complement activation pathway, (2) decreased lysis and / or decreased C3 deposition, (3) activation of antibody and complement-dependent cell cytotoxicity, (4) CH50 hemolysis, (5) decreased erythrocyte lysis, (6) decreased erythrocyte phagocytosis, (7) decreased dendritic cell infiltration, (8) inhibition of complement-mediated erythrocyte lysis, (9) decreased lymphocyte infiltration, (10) decreased macrophage infiltration, (11) decreased antibody deposition, and (12) (13) Decreased neutrophil infiltration, (14) Decreased platelet phagocytosis, (15) Improved graft survival, (16) Decreased macrophage-mediated phagocytosis, (17) Decreased autoantibody-mediated complement activation, (18) Decreased erythrocyte destruction due to transfusion, (19) Decreased erythrocyte lysis due to alloantibodies, (20) Decreased hemolysis due to transfusion, (21) Decreased alloantibody-mediated platelet lysis, (22) Improved anemia, (23) Decreased eosinophilia, (24 (25) Decreased C3 deposition on red blood cells (e.g., decreased deposition of C3b, iC3b, etc. on RBCs), (26) Decreased C3 deposition on platelets (e.g., decreased deposition of C3b, iC3b, etc. on platelets), (27) Decreased anaphylatoxin production, (28) Decreased autoantibody-mediated rash formation, (29) Decreased autoantibody-induced lupus erythematosus, (30) Decreased red blood cell destruction due to transfusion reaction, (31) Decreased platelet lysis due to transfusion reaction, (32) Obesity (32) decreased cell activation, (33) decreased mast cell histamine release, (34) decreased vascular permeability, (35) decreased complement deposition on graft endothelium, (36) B cell antibody production, (37) dendritic cell maturation, (38) T cell proliferation, (39) cytokine production, (40) microglia activation, (41) decreased Artus reaction, (42) decreased anaphylatoxin production in graft endothelium, or (42) activation of complement receptor 3 (CR3 / C3) expressing cells.

[0025] In some embodiments, CH50 hemolysis includes human, mouse, and / or rat CH50 hemolysis. In some embodiments, the antibody is capable of neutralizing at least about 50% to at least about 95% of the CH50 hemolysis. In some embodiments, the antibody is capable of neutralizing 50%, 60%, 70%, 80%, 90%, or 100% of the CH50 hemolysis. The antibody may also be capable of neutralizing at least 50% of the CH50 hemolysis at doses of less than 150 ng / ml, less than 100 ng / ml, less than 50 ng / ml, or less than 20 ng / ml.

[0026] Other in vitro assays for measuring complement activity include ELISA assays for measuring the resolution products of complement components or complexes formed during complement activation. Complement activation via the classical pathway can be measured by tracking serum C4d and C4 levels. Activation of the secondary pathway can be measured by ELISA by assessing the levels of Bb or C3bBbP complexes in circulation. In vitro antibody-mediated complement activation assays can also be used to assess the inhibition of C3a production.

[0027] The antibodies of this disclosure may be monoclonal antibodies, polyclonal antibodies, recombinant antibodies, humanized antibodies, human antibodies, chimeric antibodies, multispecific antibodies, antibody fragments thereof, or derivatives thereof. In some embodiments, the antibody is a humanized antibody.

[0028] The antibodies of this disclosure may also be antibody fragments, such as Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, diabodies, or single-chain antibody molecules. In some embodiments, the antibody fragment is a Fab fragment.

[0029] In some embodiments, the antibody is a human monoclonal antibody that can be prepared, expressed, produced or isolated by recombinant means, e.g., (a) an antibody isolated from an animal (e.g., mouse) that is transgenic or transchromosome of a human immunoglobulin gene or hybridoma prepared therefrom (as further described below), (b) an antibody isolated from host cells transformed to express the antibody, e.g., an antibody isolated from a transfectoma, (c) an antibody isolated from a recombinant combinatorial human antibody library, and (d) an antibody prepared, expressed, produced or isolated by any other means including splicing of the human immunoglobulin gene sequence to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline and / or non-germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies may undergo in vitro mutagenesis (or, if an animal transgenic to the human Ig sequence is used, in vivo somatic mutagenesis), and thus the recombinant antibody may be V H and V L The amino acid sequence of the region is human germline V H and V L These sequences are derived from and related to other sequences, but do not necessarily need to be naturally present in the human antibody germline repertoire in vivo.

[0030] In some embodiments, the antibody is a humanized and / or chimeric monoclonal antibody that can be established by immunizing rodents (e.g., mice, rats, hamsters, and guinea pigs) with (1) a natural complement component (e.g., C1q) derived from the enzymatic digestion of purified complement components from human plasma or serum, or (2) a recombinant complement component or its derivative expressed by either a eukaryotic or prokaryotic system. Other animals, such as non-human primates, transgenic mice expressing human immunoglobulins, and severe combined immunodeficiency (SCID) mice transplanted with human B lymphocytes, may be used for immunization.

[0031] Polyclonal and monoclonal antibodies are naturally produced as immunoglobulin (Ig) molecules in the immune system's response to pathogens. The dominant form in human serum, IgG1, with a concentration of approximately 8 mg / ml and a molecular weight of about 150 kDa, consists of two identical heavy chains of approximately 50 kDa and two identical light chains of approximately 25 kDa.

[0032] Hybridomas can be generated by conventional procedures, such as fusing B lymphocytes from immunized animals with myeloma cells. Anti-C1q antibodies can also be generated by screening recombinant single-stranded Fv or Fab libraries from human B lymphocytes in a phage display system. The specificity of MAb for human C1q can be tested by enzyme-linked immunosorbent assay (ELISA), Western immunoblotting, or other immunochemical techniques.

[0033] The inhibitory activity of antibodies identified in the screening process against complement activation can be evaluated by hemolysis assays using unsensitized rabbit or guinea pig RBCs for the complement subpathway, or sensitized chicken or sheep RBCs for the classical complement pathway. Hybridomas exhibiting specific inhibitory activity against the classical complement pathway are cloned by limiting dilution. The antibodies are purified for characterization regarding their specificity to human C1q using the assays described above.

[0034] definition As used herein, "a" or "an" may mean one or more. As used herein in claims (plural), when used in conjunction with the term "including", the term "a" or "an" may mean one or more. For example, a reference to "antibody" can refer to one to many antibodies. As used herein, "another" may mean at least a second or more.

[0035] As used herein, "co-administered" with another compound or composition includes simultaneous administration and / or administration at different times. Co-administration also includes administration as a co-formulation or as separate compositions, including using different dosing frequencies or intervals, and the same or different routes of administration.

[0036] The term "immunoglobulin" (Ig) is used interchangeably herein with "antibody". The term "antibody" as used herein is used in the broadest sense and specifically includes monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) formed from at least two intact antibodies, antibody fragments only when they exhibit biological activity, and antibody derivatives.

[0037] The basic four-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. V H and V L When paired together, a single antigen-binding site is formed. For the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology, 8th Ed., Daniel P. Stites, Abba I. Terr and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, page 71 and Chapter 6.

[0038] Light chains (L chains) derived from any vertebrate species can be assigned to one of two distinct types called kappa ("κ") and lambda ("λ") based on the amino acid sequence of their constant domains. Depending on the amino acid sequence of the constant domain (CH) of their heavy chains, immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM (each having a heavy chain denoted as alpha ("α"), delta ("δ"), epsilon ("ε"), gamma ("γ"), and mu ("μ"), respectively). The γ and α classes are further classified into subclasses (isotypes) based on relatively small differences in CH sequence and function. For example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known and generally described, for example, in Abbas et al., Cellular and Molecular Immunology, 4th ed. (WBSaunders Co., 2000).

[0039] A "full-length antibody" is typically a heterotetrameric glycoprotein of approximately 150,000 daltons, containing two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to the heavy chain by one covalent disulfide bond, while the number of disulfide bonds varies between heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has a variable domain (V) at one end. H ) and several subsequent constant domains. Each light chain has a variable domain (V) at one end. L It has a constant domain at the other end, the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the variable domain of the light chain is aligned with the variable domain of the heavy chain. Certain amino acid residues are thought to form an interface between the variable domain of the light chain and the variable domain of the heavy chain.

[0040] An “isolated” molecule or cell is a molecule or cell identified and separated from at least one contaminating molecule or cell that would normally be associated with the environment in which it was produced. Preferably, the isolated molecule or cell does not associate with all components associated with the environment in which it was produced. The isolated molecule or cell is in a form other than that which is found in nature or in any context. Thus, an isolated molecule is distinct from a molecule that is naturally present in a cell; an isolated cell is distinct from a cell that is naturally present in a tissue, organ, or organism. In some embodiments, the isolated molecule is the anti-C1q antibody of this disclosure. In other embodiments, the isolated cell is a host cell or hybridoma cell that produces the anti-C1q antibody of this disclosure.

[0041] An “isolated” antibody is one identified, separated, and / or recovered from components of its production environment (e.g., natural or recombinant). Preferably, the isolated polypeptide does not associate with any other contaminants from its production environment. Contaminants from its production environment, such as those resulting from recombinant transfection cells, are substances that would typically interfere with the study, diagnostic, or therapeutic use of the antibody and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In certain preferred embodiments, the polypeptide would be purified to (1) more than 95% by weight of the antibody, and in some embodiments more than 99% by weight, as determined, for example, by the Lowry method; (2) to a degree sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a spinning cup sequencer; or (3) to a degree that homogeneity is obtained by SDS-PAGE under non-reducible or reducing conditions using Coomassie blue, or preferably silver staining. Since the isolated antibody does not contain at least one component of the antibody’s natural environment, it contains the antibody in situ within recombinant T cells. However, typically, isolated polypeptides or antibodies are prepared by a process that includes at least one purification step.

[0042] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of the heavy or light chain of the antibody. The variable domains of the heavy and light chains are respectively called "V H " and "V L These domains can be referred to as "antigen-binding sites." These domains are typically the most variable parts of an antibody (compared to other antibodies of the same class) and contain antigen-binding sites.

[0043] The term "variable" refers to the fact that a particular segment of a variable domain has a significantly different sequence depending on the antibody. The V domain mediates antigen binding and defines the specificity of a particular antibody to that particular antigen. However, variability is not evenly distributed throughout the variable domain. Rather, it is concentrated in three segments called hypervariable regions (HVRs) in both the light and heavy chain variable domains. The more highly conserved portion of the variable domain is called the framework region (FR). The variable domains of the native heavy and light chains each contain four FR regions that largely adopt a beta-sheet configuration, connected by three HVRs that link beta-sheet structures and, in some cases, form loops that form part of the beta-sheet structure. The HVRs within each chain are closely linked to one another by the FR regions and, together with the HVRs of the other chain, contribute to the formation of the antibody's antigen-binding site (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, MD (1991)). The constant domain does not directly participate in antibody binding to antigens, but it exhibits various effector functions, such as the involvement of antibodies in antibody-dependent cytotoxicity.

[0044] As used herein, the term “CDR” or “complementarity-determining region” is intended to mean a discontinuous antigen-binding site found within the variable region of both heavy and light chain polypeptides. CDRs are described in Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., USDept. of Health and Human Services, “Sequences of proteins of immunological interest” (1991) (also referred to herein as Kabat 1991); Chothia et al., J. Mol. Biol. 196:901-917 (1987) (also referred to herein as Chothia 1987); and MacCallum et al., J. Mol. Biol. 262:732-745 (1996) (the definition includes duplication or subsets of amino acid residues when compared to one another). Nevertheless, any application of the definitions to refer to the CDR of an antibody or graft antibody or its variant is intended to be within the scope of the terms defined and used herein.

[0045] As used herein, the terms "CDR-L1," "CDR-L2," and "CDR-L3" refer to the first, second, and third CDRs in the light chain variable region, respectively. As used herein, the terms "CDR-H1," "CDR-H2," and "CDR-H3" refer to the first, second, and third CDRs in the heavy chain variable region, respectively. As used herein, the terms "CDR-1," "CDR-2," and "CDR-3" refer to the first, second, and third CDRs in the variable region of either chain, respectively.

[0046] The term "monoclonal antibody," as used herein, refers to an antibody obtained from a substantially homogeneous population of antibodies; that is, the individual antibodies in that population are identical except for naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation) that may be present in trace amounts. Monoclonal antibodies are highly specific and directed against a single antigenic site. In contrast to polyclonal antibody preparations, which typically contain different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous because they are typically synthesized by hybridoma culture and are free from contamination with other immunoglobulins. The modifier "monoclonal" indicates a characteristic of the antibody that it is obtained as a substantially homogeneous population of antibodies, and should not be interpreted as requiring the production of the antibody by any particular method. For example, monoclonal antibodies used in accordance with this disclosure include, for example, hybridoma methods (e.g., Kohler and Milstein, Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14(3):253-260 (1995); Harlow et al., Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, NY, 1981)), recombinant DNA methods (e.g., see U.S. Patent Nos. 4,816, 567), and phage display techniques (e.g., Clackson et al., Nature, 352:624-628 (1991); Marks et al. al.,J.Mol.Biol.222:581-597(1992);Sidhu et al.,J.Mol.Biol.338(2):299-310(2004);Lee et al.,J.Mol.Biol.340(5):1073-1093(2004);Fellouse,Proc.Nat'l Acad.Sci.See USA 101(34):12467-472(2004); and Lee et al., J. Immunol Methods 284(1-2):119-132(2004), and techniques for generating human or human-like antibodies in animals having a human immunoglobulin locus or part or all of a human gene encoding a human immunoglobulin sequence (e.g., WO1998 / 24893; WO1996 / 34096; WO1996 / 33735; WO1991 / 10741; Jakobovits et al., Proc. Nat'l Acad. Sci. USA 90:2551(1993); Jakobovits et al., Nature 362:255-258(1993); Bruggemann et al., Year in Immunol. 7:33 (1993); US Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016; Marks et al., Bio / Technology 10:779-783 (1992); Lonberg et al., Nature 368:856-859 (1994); Morrison, Nature 368:812-813 (1994); Fishwild et al., Nature Biotechnol. 14:845-851 (1996); Neuberger, Nature Biotechnol. 14:826 (1996); and Lonberg and It can be generated by a variety of techniques, including Huszar, Intern. Rev. Immunol. 13:65-93 (1995).

[0047] The terms "full-length antibody," "intact antibody," and "complete antibody" are used interchangeably to refer to an antibody in substantially intact form, as opposed to an antibody fragment or antibody derivative. Specifically, a complete antibody includes those having heavy and light chains containing an Fc region. The constant domain may be a natural sequence constant domain (e.g., a human natural sequence constant domain) or an amino acid sequence variant thereof. In some cases, an intact antibody may have one or more effector functions.

[0048] An antibody “antibody fragment,” “antigen-binding fragment,” or “functional fragment” is a portion of an intact antibody, preferably containing the F region of the intact antibody that retains or has antigen-binding and / or variable regions or modified FcR-binding ability. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; and linear antibodies (see U.S. Patent 5,641,870, Example 2; Zapata et al., Protein Eng. 8(10):1057-1062 (1995)). Additional examples of antibody fragments include antibody derivatives, such as single-chain antibody molecules, monovalent antibodies, and multispecific antibodies formed from antibody fragments.

[0049] An "antibody derivative" is any construct that includes the antigen-binding region of an antibody. Examples of antibody derivatives include single-chain antibody molecules formed from antibody fragments, monovalent antibodies, and multispecific antibodies.

[0050] Papain digestion of the antibody produces two identical antigen-binding fragments called "Fab" fragments and one residual "Fc" fragment, denoted to reflect its ability to readily crystallize. The Fab fragments consist of the entire light chain, as well as the variable region domain (V) of the heavy chain. H ), as well as the first constant domain of one heavy chain (C H 1) consists of each Fab fragment, which is monovalent with respect to antigen binding, i.e., has a single antigen-binding site. Pepsin treatment of the antibody yields a single large F(ab')2 fragment, which roughly corresponds to two disulfide-bonded Fab fragments with different antigen-binding activities and can still crosslink antigens. The Fab' fragment is C H It differs from the Fab fragment by having several additional residues at the carboxyl terminus of one domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the herein designation for Fab' fragments in which the cysteine ​​residue(s) of the constant domain have a free thiol group. The F(ab')2 antibody fragment was originally generated as a pair with a Fab' fragment having a hinged cysteine ​​in between. Other chemical couplings of the antibody fragments are also known.

[0051] The Fc fragment contains the carboxyl ends of both H chains held together by a disulfide. The effector function of the antibody is determined by the sequence in the Fc region, which is also recognized by an Fc receptor (FcR) found in certain cell types.

[0052] In this specification, the term “Fc region” is used to define the C-terminal region of an immunoglobulin heavy chain, and includes both native and variant Fc regions. While the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is typically defined as extending from the amino acid residue at position Cys226 or from Pro230 to its carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) may be removed, for example, during antibody production or purification, or by recombination of the nucleic acid encoding the antibody heavy chain. Thus, a composition of intact antibodies may include an antibody population from which all K447 residues have been removed, an antibody population without any removed K447 residues, and an antibody population having a mixture of antibodies with and without K447 residues. Native Fc regions suitable for use in the antibodies of this disclosure include human IgG1, IgG2, IgG3, and IgG4.

[0053] The "natural sequence Fc region" contains amino acid sequences identical to those found in naturally occurring Fc regions. Natural sequence human Fc regions include natural sequence human IgG1 Fc regions (non-A and A allotypes), natural sequence human IgG2 Fc regions, natural sequence human IgG3 Fc regions, and natural sequence human IgG4 Fc regions, as well as their naturally occurring variants.

[0054] A "variant Fc region" includes an amino acid sequence different from that of the natural sequence Fc region by at least one amino acid modification, preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution compared to the natural sequence Fc region or the parent polypeptide Fc region, for example, about 1 to about 10 amino acid substitutions, and preferably about 1 to about 5 amino acid substitutions, in the natural sequence Fc region or the parent polypeptide Fc region. The variant Fc region as described herein preferably has at least about 80% homology with the natural sequence Fc region and / or the parent polypeptide Fc region, most preferably at least about 90% homology, and more preferably at least about 95% homology.

[0055] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. The preferred FcR is the natural sequence human FcR. Furthermore, preferred FcRs include receptors of the FcγRI, FcγRII, and FcγRIII subclasses that bind to IgG antibodies (gamma receptors), and include allelic variants and alternative splicing forms of these receptors. The FcγRII receptor includes FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences that differ mainly in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif ("ITAM") in its cytoplasmic domain. The inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibitory motif ("ITIM") in its cytoplasmic domain. (See, for example, M. Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs have been reviewed in Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are included in the term “FcR” as used herein. FcRs may increase the serum half-life of antibodies.

[0056] The in vivo binding of human FcRn-high affinity binding polypeptides to FcRn and their serum half-lives can be assayed, for example, in transgenic mice or transfected human cell lines expressing human FcRn, or in primates administered with polypeptides containing a variant Fc region. WO2004 / 42072 (Presta) describes antibody variants with improved or reduced binding to FcR. See, for example, Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001).

[0057] "Fv" is the smallest antibody fragment containing both a complete antigen recognition site and an antigen binding site. This fragment consists of a dimer in which one heavy chain variable domain and one light chain variable domain are tightly bound noncovalently. The folding of these two domains creates six hypervariable loops (three from the H chain and three from the L chain) that provide amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of Fv containing only three antigen-specific HVRs) has the ability to recognize and bind to an antigen, although with lower affinity than the full binding site.

[0058] "Single-chain Fv," also abbreviated as "sFv" or "scFv," is an antibody fragment containing VH and VL antibody domains bound to a single polypeptide chain. Preferably, the sFv polypeptide is V H and V L The sFv further contains polypeptide linkers between its domains, enabling it to form the structure desired for antigen binding. For an overview of sFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0059] The term "diabody" refers to the process of achieving inter-chain V-domain pairing, rather than intra-chain pairing, thereby obtaining a bivalent fragment, i.e., a fragment having two antigen-binding sites. H Domain and V L This refers to a small antibody fragment prepared by constructing an sFv fragment (see previous paragraph) using a short linker (approximately 5-10 residues) between the domain and the sFv. A bispecificity diabody is a V of two antibodies. H and V LIt is a heterodimer of two "cross-over" sFv fragments located in polypeptide chains with different domains. The diabody is described in more detail, for example, EP 404,097;WO 1993 / 011161;WO / 2009 / 121948;WO / 2014 / 191493;Hollinger et al., Proc. Nat'l Acad. Sci. USA 90:6444-48 (1993).

[0060] As used herein, “chimeric antibody” refers to an antibody (immunoglobulin) in which a portion of the heavy chain and / or light chain is identical or homologous to a corresponding sequence in an antibody of a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to a corresponding sequence in an antibody of a different species or belonging to a different antibody class or subclass, as well as fragments of such antibodies, insofar as they exhibit the desired biological activity (U.S. Patent No. 4,816,567; Morrison et al., Proc. Nat'l Acad. Sci. USA, 81:6851-55 (1984)). The chimeric antibodies used herein include PRIMATIZED® antibodies, in which the antigen-binding region of the antibody is derived, for example, from an antibody produced by immunizing a macaque monkey with the antigen of interest. As used herein, “humanized antibody” is a subset of “chimeric antibody.”

[0061] The "humanized" form of a non-human (e.g., mouse) antibody is a chimeric antibody containing a minimal sequence derived from a non-human immunoglobulin. In some embodiments, the humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's HVR are replaced with residues from the HVR of a non-human species (donor antibody), such as mouse, rat, rabbit, or non-human primate, having the desired specificity, affinity, and / or capabilities. In some examples, FR residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, the humanized antibody may contain residues not found in either the recipient antibody or the donor antibody. These modifications may be made to further improve the antibody's performance, such as binding affinity. Generally, humanized antibodies will contain substantially all of at least one, typically two, variable domains, where all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin sequence, and all or substantially all of the FR region corresponds to those of a human immunoglobulin sequence, although the FR region may contain one or more individual FR residue substitutions that improve antibody performance such as binding affinity, isomerization, and immunogenicity. The number of these amino acid substitutions in the FR is typically 6 or less in the H chain and 3 or less in the L chain. Humanized antibodies also optionally contain at least a portion of the immunoglobulin constant region (Fc), typically at least a portion of human immunoglobulin. For further details, see, for example, Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See, for example, Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1:105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Patent Nos. 6,982,321 and 7,087,409.

[0062] A “human antibody” is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human, and / or one prepared using any of the techniques for producing human antibodies disclosed herein. This definition of a human antibody specifically excludes humanized antibodies containing non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J.Mol.Biol., 227:381 (1991); Marks et al., J.Mol.Biol., 222:581 (1991). The method described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p.77 (1985); Boerner et al., J.Immunol., 147(1):86-95 (1991) is also available for the preparation of human monoclonal antibodies. See also van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5:368-74 (2001). Human antibodies can be prepared by administering an antigen to transgenic animals, such as immunized xenomous mice, which have been modified to produce such antibodies in response to antigen attack, but whose endogenous gene locus has been deactivated (see, for example, U.S. Patent Nos. 6,075,181 and 6,150,584 for XENOMOUSE® technology). See also Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) for human antibodies produced by human B-cell hybridoma technology, for example.

[0063] As used herein, the terms “hypervariable region,” “HVR,” or “HV” refer to regions of antibody variable domains whose sequences are hypervariable and / or form structurally defined loops. Generally, antibodies contain six HVRs, three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). In natural antibodies, H3 and L3 exhibit the highest diversity among these six HVRs, and H3 in particular is thought to play a unique role in conferring superior specificity to antibodies. See, for example, Xu et al., Immunity 13:37-45 (2000); Johnson and Wu in Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, NJ, 2003). In practice, naturally occurring camelid antibodies consisting only of heavy chains are functional and stable in the absence of light chains. For example, see Hamers-Casterman et al., Nature 363:446-448 (1993) and Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0064] Several HVR descriptions are used and incorporated herein. Kabat complementarity-determining regions (CDRs) are HVRs based on sequence variability and are the most commonly used (Kabat et al., above). Chothia, on the other hand, refers to the location of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). AbM HVRs represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. "Contact" HVRs are based on analysis of available complex crystal structures. The residues from which each of these HVRs originate are listed below.

[0065] [Table 1]

[0066] HVR may include “extended HVR” as follows: 24–36 or 24–34 (L1), 46–56 or 50–56 (L2), and 89–97 or 89–96 (L3) in VL, and 26–35 (H1), 50–65 or 49–65 (preferred embodiment) (H2), and 93–102, 94–102, or 95–102 (H3) in VH. Variable domain residues are numbered according to Kabat et al. above for each of these definitions of extended HVR.

[0067] A “framework” or “FR” residue is a variable domain residue other than an HVR residue as defined herein.

[0068] The terms “Kabat-like variable domain residue numbering” or “Kabat-like amino acid position numbering,” and their variations, refer to the numbering system used for the heavy-chain or light-chain variable domains of antibody edits in Kabat et al. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortenings or insertions of the FR or HVR of the variable domain. For example, the heavy-chain variable domain may contain a single amino acid insertion after H2 residue 52 (residue 52a according to Kabat) and a residue inserted after heavy-chain FR residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). The Kabat numbering of residues can be determined for a given antibody by matching the homologous region between the antibody sequence and the “standard” Kabat-numbered sequence.

[0069] The Kabat numbering system is generally used when referring to residues within the variable domain (roughly residues 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The "EU numbering system" or "EU index" is generally used when referring to residues within the constant region of the immunoglobulin heavy chain (e.g., the EU index reported by Kabat et al. above). "EU index as found in Kabat" refers to the residue numbering of human IgG1 EU antibodies. Unless otherwise stated herein, references to residue numbers in the variable domain of an antibody mean residue numbering according to the Kabat numbering system. Unless otherwise stated herein, references to residue numbers in the constant domain of an antibody mean residue numbering according to the EU numbering system (e.g., see U.S. Patent Publication No. 2010-280227).

[0070] When used herein, “acceptor human framework” is a framework comprising the amino acid sequence of a VL framework or VH framework derived from a human immunoglobulin framework or human consensus framework. An acceptor human framework “derived from” a human immunoglobulin framework or human consensus framework may comprise the same amino acid sequence, or it may comprise existing amino acid sequence variations. In some embodiments, the number of existing amino acid variations is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. If existing amino acid variations are present in VH, preferably those variations are present in only three, two, or one of positions 71H, 73H, and 78H, for example, the amino acid residues at those positions may be 71A, 73T, and / or 78A. In some embodiments, the VL acceptor human framework is identical in sequence to the VL human immunoglobulin framework sequence or human consensus framework sequence.

[0071] The "Human Consensus Framework" is a framework that represents the most commonly occurring amino acid residues in the selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is performed from subgroups of variable domain sequences. Generally, the sequence subgroups are those found in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991). Examples include those for VL, where the subgroups may be subgroups Kappa I, Kappa II, Kappa III, or Kappa IV, as described in Kabat et al. Furthermore, for VH, the subgroups may be subgroups I, II, or III, as described in Kabat et al.

[0072] An "amino acid modification" at a specified location refers to the substitution or deletion of a specified residue, or the insertion of at least one amino acid residue adjacent to the specified residue. An "adjacent" insertion to a specified residue means an insertion within one or two residues. The insertion may be at the N-terminus or C-terminus of the specified residue. The preferred amino acid modification in this specification is substitution.

[0073] Affinity-matured antibodies have one or more changes in one or more of their HVRs, and these changes result in improved affinity of the antibody to the antigen compared to parent antibodies that do not have those changes. In some embodiments, affinity-matured antibodies have nanomolar or even picomolar affinity for the target antigen. Affinity-matured antibodies are produced by procedures known in the art. For example, Marks et al., Bio / Technology 10:779-783 (1992) describe affinity maturation by VH and VL domain shuffling. Random mutagenesis of HVR and / or framework residues is described, for example, in Barbas et al. Proc Nat.Acad.Sci.USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Yelton et al. J.Immunol.155:1994-2004 (1995); Jackson et al., J.Immunol.154(7):3310-9 (1995); and Hawkins et al., J.Mol.Biol.226:889-896 (1992).

[0074] As used herein, the terms “specifically recognize” or “specifically bind” refer to measurable and reproducible interactions, such as attraction or binding, between a target and an antibody that determine the presence of the target in the presence of a heterogeneous population of molecules, including biological molecules. For example, an antibody that specifically or preferentially binds to a target or epitope is an antibody that binds to that target or epitope with higher affinity, avidity, more readily, and / or for a longer duration than when it binds to other targets or other epitopes of a target. For example, it is also understood that an antibody (or site) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. Thus, “specific binding” or “preferential binding” does not necessarily require (but may include) exclusive binding. An antibody that specifically binds to a target is at least about 10 3 M -1 or 10 4 M -1 , sometimes about 10 5 M -1 or 10 6 M -1 In other examples, approximately 10 6 M -1 or 10 7 M -1 , about 10 8 M -1 ~10 9 M -1 , or about 10 10 M -1 ~10 11 M -1 It may have a binding constant of or greater. A variety of immunoassay formats can be used to select antibodies with specific immunoreactivity to a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select monoclonal antibodies with specific immunoreactivity to a protein. For a description of immunoassay formats and conditions that may be used to determine specific immunoreactivity, see, for example, Harlow and Lane (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York.

[0075] "Identity," as used herein, indicates that amino acid residues are identical between sequences at any particular position in the aligned sequences. "Similarity," as used herein, indicates that amino acid residues are of a similar type between sequences at any particular position in the aligned sequences. For example, leucine may be used instead of isoleucine or valine. Other amino acids that are often substituted for each other include: - Phenylalanine, tyrosine, and tryptophan (amino acids with aromatic side chains); - Lysine, arginine, and histidine (amino acids with basic side chains); - Aspartate and glutamate (amino acids with acidic side chains); -Asparagine and glutamine (amino acids with amide side chains); and - This includes, but is not limited to, cysteine ​​and methionine (amino acids with sulfur-containing side chains).

[0076] The degree of identity and similarity can be easily calculated (see, for example, Computational Molecular Biology, Lesk, AM, ed., Oxford University Press, New York, 1988; Biocomputing. Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991).

[0077] As used herein, “interaction” between a complement protein and a second protein includes, but is not limited to, protein-protein interactions, physical interactions, chemical interactions, binding, covalent bonds, and ionic bonds. As used herein, an antibody “inhibits interaction” between two proteins if it disrupts, reduces, or completely eliminates the interaction between the two proteins. The antibody or fragment thereof “inhibits interaction” between two proteins if it binds to one of the two proteins.

[0078] A “blocking” antibody, “antagonist” antibody, “inhibitory” antibody, or “neutralizing” antibody is an antibody that inhibits or reduces one or more biological activities of the antigen to which it binds, such as interactions with one or more proteins. In some embodiments, a blocking antibody, antagonist antibody, inhibitory antibody, or “neutralizing” antibody substantially or completely inhibits one or more biological activities or interactions of the antigen.

[0079] The term "inhibitor" refers to a compound that has the ability to inhibit the biological function of a target biomolecule, such as mRNA or a protein, whether by reducing the activity or expression of the target biomolecule. Inhibitors can be antibodies, small molecules, or nucleic acid molecules. The term "antagonist" refers to a compound that binds to a receptor and blocks or attenuates the receptor's biological response. The term "inhibitor" can also refer to an "antagonist."

[0080] The "effector function" of an antibody refers to the biological activity resulting from the antibody's Fc region (either the natural sequence Fc region or the amino acid sequence variant Fc region), and this varies depending on the antibody isotype.

[0081] As used herein, the term “affinity” is the equilibrium constant for the reversible binding of two substances (e.g., an antibody and an antigen), and is expressed as the dissociation constant (KD). The affinity may be at least 1-fold higher, at least 2-fold higher, at least 3-fold higher, at least 4-fold higher, at least 5-fold higher, at least 6-fold higher, at least 7-fold higher, at least 8-fold higher, at least 9-fold higher, at least 10-fold higher, at least 20-fold higher, at least 30-fold higher, at least 40-fold higher, at least 50-fold higher, at least 60-fold higher, at least 70-fold higher, at least 80-fold higher, at least 90-fold higher, at least 100-fold higher, or at least 1,000-fold higher or more than that. The affinity of an antibody for a target protein may be, for example, about 100 nanomolar (nM) to about 0.1 nM, about 100 nM to about 1 picomolar (pM), or about 100 nM to about 1 femtomole (fM) or more. As used herein, the term "avidity" refers to the resistance of a complex of two or more substances to dissociation after dilution. The terms "immunoreactivity" and "preferentially binding" are used interchangeably herein with respect to antibodies and / or antigen-binding fragments.

[0082] The term "binding" refers to the direct association between two molecules by covalent, electrostatic, hydrophobic, and ionic and / or hydrogen bonding interactions, including interactions such as salt and water bridges. For example, a target anti-C1q antibody specifically binds to an epitope within the complement C1q protein. "Specific binding" refers to at least approximately 10 -7 M or more, for example, 5 x 10 -7 M, 10 -8 M, 5×10 -8 This refers to binding with affinity M and above. "Non-specific binding" is approximately 10 -7 Affinity less than M, for example, 10 -6 M, 10 -5 M, 10 -4 This refers to binding based on affinity, such as M.

[0083] The term "k" onWhen used herein, " is intended to refer to the rate constant for the association of antibodies with an antigen.

[0084] The term "k" off When used herein, " is intended to refer to the rate constant for the dissociation of an antibody from an antibody / antigen complex.

[0085] The term “K D When used herein, " is intended to refer to the equilibrium dissociation constant of antibody-antigen interactions.

[0086] As used herein, with respect to peptide, polypeptide, or antibody sequences, “amino acid sequence identity percentage (%)” and “homology” refer to the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence, after the sequences have been aligned and gaps introduced as necessary to achieve the maximum sequence identity percentage, without considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining the amino acid sequence identity percentage can be achieved in various ways within the art, for example, using publicly available computer software, such as BLAST, BLAST-2, ALIGN, or MEGALIGN® (DNASTAR) software. A person skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm known in the art necessary to achieve the maximum alignment over the entire length of the sequences being compared.

[0087] The term "biological sample" encompasses a diverse range of sample types obtained from an individual and may be used in diagnostic or monitoring assays. Its definition includes blood and other liquid samples of biological origin, solid tissue samples, e.g., biopsy samples or tissue cultures or cells and their offspring derived therefrom. It also includes samples that have been manipulated in any way after their procurement, for example, by treatment, solubilization, or concentration with reagents for certain components, such as polynucleotides. The term "biological sample" encompasses clinical samples and also includes cultured cells, cell supernatants, cell lysates, serum, plasma, biological fluids, and tissue samples. The term "biological sample" includes urine, saliva, cerebrospinal fluid, interstitial fluid, ocular fluid, synovial fluid, blood fractions, e.g., plasma and serum. The term "biological sample" also includes solid tissue samples, tissue culture samples, and cell samples.

[0088] An "isolated" nucleic acid molecule is a nucleic acid molecule that has been identified and separated from at least one contaminating nucleic acid molecule that is normally associated with the environment in which it was produced. Preferably, the isolated nucleic acid does not associate with any components associated with the environment in which it was produced. Isolated nucleic acid molecules encoding polypeptides and antibodies as used herein are in forms or circumstances other than those found in nature. Therefore, isolated nucleic acid molecules are distinguished from nucleic acids encoding any polypeptides and antibodies as used herein that are naturally present in cells.

[0089] The term “vector,” as used herein, is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it is ligated. One type of vector is a “plasmid,” which refers to a circular double-stranded DNA to which an additional DNA segment can be ligated. Another type of vector is a phage vector. Another type of vector is a viral vector, in which case an additional DNA segment can be ligated into the viral genome. Certain vectors are capable of self-replication in the host cell into which they are introduced (e.g., bacterial vectors with bacterial origins of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors), upon introduction into a host cell, can be integrated into the host cell's genome and thereby replicate with the host genome. Furthermore, certain vectors are capable of directing the expression of a gene to which they are functionally ligated. Such vectors are referred herein as “recombinant expression vectors,” or simply “expression vectors.” Expression vectors that are typically useful in recombinant DNA technology are often in the form of plasmids. In this specification, “plasmid” and “vector” may be used interchangeably, as plasmids are the most commonly used form of vector.

[0090] As used interchangeably herein, “polynucleotide” or “nucleic acid” refers to a polymer of nucleotides of any length, including DNA and RNA. Nucleotides may be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or analogs thereof, or any substrate that can be incorporated into the polymer by DNA or RNA polymerase or by a synthetic reaction. Polynucleotides may include modified nucleotides, e.g., methylated nucleotides and their analogs. Modifications to the nucleotide structure, if present, may be conjugated before or after the assembly of the polymer. Nucleotide sequences may be separated by non-nucleotide components. Polynucleotides may include post-synthesis-generated modifications, e.g., conjugations to labels. Other types of modifications include, for example, “caps” that replace one or more naturally occurring nucleotides with analogs; internucleotide modifications, such as those by non-charged bonds (e.g., methylphosphonates, phosphotriesters, phosphoamidates, carbamates, etc.) and those by charged bonds (e.g., phosphorothioates, phosphorodithioates, etc.), such as those involving suspension sites of proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.); those involving insertors (e.g., acridine, psoralen, etc.); those involving chelating agents (e.g., metals, radioactive metals, boron, metal oxides, etc.); those involving alkylating agents; those involving modified bonds (e.g., alpha-anomeric nucleic acids, etc.); and the unmodified form of polynucleotides. Furthermore, any of the hydroxyl groups normally present in the sugar may be replaced, for example, by a phosphonic acid group or a phosphate group, protected by a standard protecting group, or activated to generate additional binding to additional nucleotides, or conjugated to a solid or semi-solid support. The 5' and 3' terminal OH groups may be phosphorylated or substituted with amines or organic capping groups of 1 to 20 carbon atoms. Other hydroxyls may also be derivatized to standard protecting groups.Polynucleotides may include analog forms of ribose or deoxyribose sugars commonly known in the art, such as 2'-O-methyl-, 2'-O-allyl, 2'-fluoro- or 2'-azidol-ribose, carbocyclic sugar analogs, α-anomeric sugars, epimeric sugars, such as arabinose, xylose or lyxose, pyranose sugars, furanose sugars, sedoheptulose, acrylate analogs, and basic nucleoside analogs, such as methylriboside. One or more phosphodiester bonds may be replaced by alternative linking groups. These alternative linking groups include, but are not limited to, embodiments in which the phosphate is replaced by P(O)S ("thioate"), P(S)S ("dithioate"), (O)NR2 ("amidate"), P(O)R, P(O)OR', CO, or CH2 ("formacetal") (wherein each R or R' is independently a substituted or unsubstituted alkyl (1-20C), aryl, alkenyl, cycloalkyl, cycloalkenyl, or aralkyl, optionally containing H or an ether (-O-) bond). Not all links in the polynucleotide need to be identical. The foregoing description applies to all polynucleotides referred to herein, including RNA and DNA.

[0091] "Host cells" include individual cells or cell cultures that may be recipients for a vector(s) for the incorporation of polynucleotide insertions. Host cells include offspring of a single host cell, which may not necessarily be identical (in morphology or genomic DNA complement) to the original parent cell due to spontaneous, spontaneous, or intentional mutation. Host cells include cells transfected in vivo with the polynucleotide(s) of this disclosure.

[0092] As used herein, “carrier” includes pharmaceutically acceptable carriers, excipients, or stabilizers that are non-toxic to cells or mammals to which they are exposed at the dosage and concentration used. Often, physiologically acceptable carriers are aqueous pH buffers. Examples of physiologically acceptable carriers include buffers, e.g., phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, e.g., serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, e.g., EDTA; sugar alcohols, e.g., mannitol or sorbitol; salt-forming counterions, e.g., sodium; and / or nonionic surfactants, e.g., TWEEN®, polyethylene glycol (PEG), and PLURONICS®.

[0093] The term "prevent" is recognized in the relevant technical field and, when used in relation to conditions such as eye diseases (e.g., glaucoma or age-related macular degeneration such as AMD including geographic atrophy) or related symptoms, refers to untreated patients.

[0094] The term “subject,” as used herein, refers to a living mammal and may be used interchangeably with the term “patient.” Examples of mammals include any member of the mammalian class: humans, non-human primates, e.g., chimpanzees, and other apes and monkey species; pasture animals, e.g., cattle, horses, sheep, goats, and pigs; domestic animals, e.g., rabbits, dogs, and cats; and laboratory animals, e.g., rodents, e.g., rats, mice, and guinea pigs. The term does not indicate a specific age or sex.

[0095] As used herein, the terms “to treat” or “treatment” include reducing, cessating or reversing the symptoms, clinical signs, or underlying pathology of a condition, thereby stabilizing or improving the condition in question, or reducing the likelihood that the condition in question would worsen to the same extent as if the subject had not received treatment.

[0096] With respect to the treatment method in question, the term “therapeutic effective dose” of a compound refers to the amount of the compound(s) in a preparation that, when administered as part of a desired drug regimen (for mammals, preferably humans), is necessary to treat a disease or condition, or to alleviate symptoms, improve a condition, or delay the onset of a disease or condition, in accordance with clinically acceptable standards for cosmetic purposes, at a reasonable benefit / risk ratio applicable to any medical treatment. The therapeutic effective dose as used herein may vary depending on factors such as the patient’s disease state, age, sex, and weight, as well as the antibody’s ability to produce a desired response in the individual.

[0097] As used herein, an individual “at risk” of developing a particular disease, disorder, or condition does not have to be present with a detectable disease or symptoms of the disease, nor does it have to be present with a detectable disease or symptoms of the disease prior to the treatment method described herein. “At risk” means that an individual has one or more risk factors, which are measurable parameters that correlate with the development of a particular disease, disorder, or condition as known in the art. An individual having one or more of these risk factors is more likely to develop a particular disease, disorder, or condition than an individual not having one or more of these risk factors.

[0098] "Chronic" administration refers to the continuous administration of a drug(s) to maintain an initial therapeutic effect (activity) over a long period, as opposed to an acute approach. "Intermittent" administration refers to a treatment that is not administered continuously without interruption, but is effectively periodic / regular.

[0099] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention belongs. Any methods and substances similar or equivalent to those described herein may also be used in the practice or testing of the present invention, but preferred methods and substances are described below. All publications listed herein are incorporated herein by reference, and the methods and / or substances cited in these publications include, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual 3rd edition (2001), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (FMAusubel, et al. eds., (2003)); the series Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (MJ MacPherson, BD Hames and GRTaylor eds. (1995)), Harlow and Lane, eds. (1988); Antibodies, A Laboratory Manual, and Animal Cell Culture (RI Freshney, ed. (1987)); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook(JECellis,ed.,1998)Academic Press;Animal Cell Culture(RIFreshney),ed.,1987);Introduction to Cell and Tissue Culture(JPMather and PERoberts,1998)Plenum Press;Cell and Tissue Culture:Laboratory Procedures(A.Doyle,JBGriffiths,and DGNewell, eds., 1993-8) J. Wiley and Sons; Handbook of Experimental Immunology (DMWeir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JMMiller and MP Calos, eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (JEColigan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (CA Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty., ed., IRL Press,1988-1989);Monoclonal Antibodies:A Practical Approach(P.Shepherd and This paper discloses and describes widely used methodologies found in C. Dean, eds., Oxford University Press, 2000; Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane, Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and JDCapra, eds., Harwood Academic Publishers, 1995); and Cancer: Principles and Practice of Oncology (VT DeVita et al., eds., JBLippincott Company, 1993).

[0100] Anti-complement C1q antibody The anti-C1q antibodies disclosed herein are potent inhibitors of C1q and can be administered for continuous inhibition of C1q function over any period of time, and then optionally discontinued to allow for the restoration of normal C1q function at a point where their activity may be important. Results obtained in animal studies with the anti-C1q antibodies disclosed herein can be readily translated to clinical applications using humanized or human antibodies, as well as fragments and / or derivatives thereof.

[0101] C1q is a large 460kDa multimeric protein composed of 18 polypeptide chains (6 C1q A chains, 6 C1q B chains, and 6 C chains). The C1r and C1s complement proteins bind to the C1q tail region to form the C1 complex (C1qr2s2).

[0102] The antibodies of this disclosure specifically recognize complement factor C1q and / or C1q in the C1 complex of the classical complement activation pathway. The bound complement factor may originate from any mammalian organism, including, but not limited to, humans, mice, rats, rabbits, monkeys, dogs, cats, cattle, horses, camels, sheep, goats, or pigs, as well as any organism having a complement system.

[0103] As used herein, “C1 complex” means a protein complex that may include, but is not limited to, one C1q protein, two C1r proteins, and two C1s proteins (e.g., C1qr 2 s 2 ) refers to.

[0104] The anti-C1q antibodies disclosed herein can inhibit C1 complex formation.

[0105] As used herein, "complement factor C1q" refers to both the wild-type sequence and the naturally occurring variant sequence.

[0106] A non-limiting example of complement factor C1q recognized by the antibodies of this disclosure is human C1q comprising three polypeptide chains A, B, and C: C1q, Chain A (homo sapiens), Accession number, Protein Database: NP_057075.1; GenBank number: NM_015991: >gi|7705753|ref|NP_057075.1|Complement C1q Subcomponent subunit A precursor [Homo sapiens] (Sequence ID 1) MEGPRGWLVLCVLAISLASMVTEDLCRAPDGKKGEAGRPGRRGRPGLKGEQGEPGAPGIRTGIQGLKGDQGEPGPSGNPGKVGYPGPSGPLGARGIPGIKGTKGSPGNIKDQPRPAFSAIRRN PPMGGNVVIFDTVITNQEEPYQNHSGRFVCTVPGYYYYFTFQVLSQWEICLSIVSSSSRGQVRRSLGFCDTTNKGLFQVVSGGMVLQLQQGDQVWVEKDPKKGHIYQGSEADSVFSGFLIFPSA.

[0107] C1q, chain B (homo sapiens), accession number protein Database: NP_000482.3; GenBank number: NM_000491.3: >gi|87298828|ref|NP_000482.3|Complement Clq Subcomponent subunit B precursor [Homo sapiens] (Sequence 2) MMMKIPWGSIPVLMLLLLGLIDISQAQLSCTGPPAIPGIPGIPGTPGPDGQPGTPGIKGEKGLPGLAGDHGEFGEKGDPGIPGNPGKVGPKGPMGPKGGPGAPGAPGPKGESGDYKATQKIAFSAT RTINVPLRRDQTIRFDHVITNMNNNYEPRSGKFTCKVPGLYYFTYHASSRGNLCVNLMRGRERAQKVVTFCDYAYNTFQVTTGGMVLKLEQGENVFLQATDKNSLLGMEGANSIFSGFLLFPDMEA.

[0108] C1q, chain C (homo sapiens), accession number protein Database: NP_001107573.1; GenBank number: NM_001114101.1: >gi|166235903|ref|NP_001107573.1|Complement C1q Subcomponent subunit C precursor [Homo sapiens] (Sequence ID 3) MDVGPSSLPHLGLKLLLLLLLLPLRGQANTGCYGIPGMPGLPGAPGKDGYDGLPGPKGEPGIPAIPGIRGPKGQKGEPGLPGHPGKNGPMGPGMPGVPGPMGIPGEPGEEGRYKQKFQSVFT VTRQTHQPPAPNSLIRFNAVLTNPQGDYDTSTGKFTCKVPGLYYFVYHASHTANLCVLLYRSGVKVVTFCGHTSKTNQVNSGGVLLRLQVGEEVWLAVNDYYDMVGIQGSDSVFSGFLLFPD.

[0109] Therefore, the anti-C1q antibody of this disclosure may bind to polypeptide chain A, polypeptide chain B, and / or polypeptide chain C of the C1q protein. In some embodiments, the anti-C1q antibody of this disclosure binds to polypeptide chain A, polypeptide chain B, and / or polypeptide chain C of human C1q or its homolog, e.g., mouse, rat, rabbit, monkey, dog, cat, cattle, horse, camel, sheep, goat, or pig C1q. In some embodiments, the anti-C1q antibody is a human antibody, a humanized antibody, a chimeric antibody, or a fragment thereof or a derivative thereof. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is an antibody fragment such as a Fab fragment.

[0110] All sequences cited in the following 20 paragraphs are incorporated herein by reference from U.S. Patent No. 9,708,394 (which is incorporated herein by reference with respect to the antibodies and related compositions disclosed therein).

[0111] Variable domain sequences of the light and heavy chains of antibody M1 (Mab1) Using standard techniques, the nucleic acid and amino acid sequences encoding the light chain variable and heavy chain variable domains of antibody M1 were determined. The amino acid sequence of the light chain variable domain of antibody M1 is:

[0112] The filename is TIFF2023551734000002.tif19170.

[0113] The hypervariable regions (HVRs) of the light chain variability domains are indicated in bold and underlined text. In some embodiments, HVR-L1 of the M1 light chain variability domain has the sequence RASKSINKYLA (SEQ ID NO: 5), HVR-L2 of the M1 light chain variability domain has the sequence SGSTLQS (SEQ ID NO: 6), and HVR-L3 of the M1 light chain variability domain has the sequence QQHNEYPLT (SEQ ID NO: 7).

[0114] The amino acid sequence of the heavy chain variable domain of antibody M1 is:

[0115] The filename is TIFF2023551734000003.tif26170.

[0116] The hypervariable region (HVR) of the heavy chain variable domain is indicated in bold and underlined text. In some embodiments, HVR-H1 of the M1 heavy chain variable domain has the sequence GYHFTSYWMH (SEQ ID NO: 9), HVR-H2 of the M1 heavy chain variable domain has the sequence VIHPNSGSINYNEKFES (SEQ ID NO: 10), and HVR-H3 of the M1 heavy chain variable domain has the sequence ERDSTEVLPMDY (SEQ ID NO: 11).

[0117] The nucleic acid sequence encoding the light chain variable domain is, GATGTCCAGATAACCCAGTCTCCATCTTATCTTGCTGCATCTCCTGGAGAAACCATTACTATTAATTGCAGGGCAAGTAAGAGCATTAACAAATATTTAGCCTGGTATCAAGAGAAACCTGGGAAAACTAATAAGCTTCTTATCTACTCTGGATCCACTTTGCAATC It was determined that TGGAATTCCATCAAGGTTCAGTGGCAGTGGATCTGGTACAGATTTCACTCTCACCATCAGTAGCCTGGAGCCTGAAGATTTTGCAATGTATTACTGTCAACAACATAATGAATACCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 12).

[0118] The nucleic acid sequence encoding the heavy chain variable domain is, CAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGTTGTCCTGCAAGTCTTCTGGCTACCATTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAGTGATTCATCCTAATAGTGGTAGTATTAACTACAATGAGAA It was determined that GTTCGAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCGGCGGTCTATTATTGTGCAGGAGAGAGAGATTCTACGGAGGTTCTCCCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 13).

[0119] Deposit of materials The following materials have been deposited in accordance with the Budapest Convention with the American Type Culture Collection, ATCC Patent Depository, 10801 University Blvd., Manassas, Va. 20110-2209, USA (ATCC):

[0120] [Table 2]

[0121] A hybridoma cell line (mouse hybridoma C1qM1 7788-1(M)051613) that produces the M1 antibody has been deposited with ATCC under the condition that the cultures will be available during the pendency of the patent application and for 30 years, or 5 years from the latest claim, or for the longer of the patent's term. The deposited material will be replaced if it becomes unviable during that period. The deposited material is available as required by foreign patent law in the country where the corresponding application of this application or its descendants is filed. However, it should be understood that the availability of the deposited material does not constitute a license to practice the invention beyond the patent rights granted by government measures.

[0122] This specification discloses a method for administering an anti-C1q antibody comprising a light chain variable domain and a heavy chain variable domain. The antibody may bind to at least human C1q, mouse C1q, or rat C1q. The antibody may be a humanized antibody, a chimeric antibody, or a human antibody. The antibody may be a monoclonal antibody, an antibody fragment thereof, and / or an antibody derivative thereof. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is an antibody fragment such as a Fab fragment. The light chain variable domain comprises HVR-L1, HVR-L2, and HVR-L3 of monoclonal antibody M1 produced by the hybridoma cell line deposited under accession number PTA-120399. The heavy chain variable domain comprises HVR-H1, HVR-H2, and HVR-H3 of monoclonal antibody M1 produced by the hybridoma cell line deposited under ATCC accession number PTA-120399.

[0123] In some embodiments, the amino acid sequences of the light chain variable domain and the heavy chain variable domain include one or more of the following: SEQ ID NO: 5 for HVR-L1, SEQ ID NO: 6 for HVR-L2, SEQ ID NO: 7 for HVR-L3, SEQ ID NO: 9 for HVR-H1, SEQ ID NO: 10 for HVR-H2, and SEQ ID NO: 11 for HVR-H3.

[0124] The antibody may preferably retain HVR-L1 RASKSINKYLA (SEQ ID NO: 5), HVR-L2 SGSTLQS (SEQ ID NO: 6), and HVR-L3 QQHNEYPLT (SEQ ID NO: 7), while containing at least 85%, 90%, or 95% of the same light chain variable domain amino acid sequence as SEQ ID NO: 4. The antibody may preferably retain HVR-H1 GYHFTSYWMH (SEQ ID NO: 9), HVR-H2 VIHPNSGSINYNEKFES (SEQ ID NO: 10), and HVR-H3 ERDSTEVLPMDY (SEQ ID NO: 11), while containing at least 85%, 90%, or 95% of the same heavy chain variable domain amino acid sequence as SEQ ID NO: 8.

[0125] This specification discloses a method of administering an anti-C1q antibody that inhibits the interaction between C1q and autoantibodies. In a preferred embodiment, the anti-C1q antibody induces clearance of C1q from circulation or tissue.

[0126] In some embodiments, the anti-C1q antibody of this disclosure inhibits the interaction between C1q and C1s. In some embodiments, the anti-C1q antibody inhibits the interaction between C1q and C1r. In some embodiments, the anti-C1q antibody inhibits the interactions between C1q and C1s and between C1q and C1r. In some embodiments, the anti-C1q antibody inhibits the interaction between C1q and another antibody, e.g., an autoantibody. In preferred embodiments, the anti-C1q antibody causes clearance of C1q from circulation or tissue. In some embodiments, the anti-C1q antibody inhibits each interaction in stoichiometric ratios of 2.5:1; 2.0:1; 1.5:1; or less than 1.0:1. In some embodiments, the C1q antibody inhibits interactions such as the C1q-C1s interaction with C1q and the anti-C1q antibody at approximately equimolar concentrations. In other embodiments, the anti-C1q antibody is expressed in ratios of less than 20:1; less than 19.5:1; less than 19:1; less than 18.5:1; less than 18:1; less than 17.5:1; less than 17:1; less than 16.5:1; less than 16:1; less than 15.5:1; less than 15:1; less than 14.5:1; less than 14:1; less than 13.5:1; less than 13:1; less than 12.5:1; less than 12.1; less than 11.5:1; 11:1 The antibody binds to C1q in a stoichiometric ratio of less than 10.5:1, less than 10:1, less than 9.5:1, less than 9:1, less than 8.5:1, less than 8.1, less than 7.5:1, less than 7:1, less than 6.5:1, less than 6:1, less than 5.5:1, less than 5:1, less than 4.5:1, less than 4:1, less than 3.5:1, less than 3:1, less than 2.5:1, less than 2.0:1, less than 1.5:1, or less than 1.0:1. In certain embodiments, the anti-C1q antibody binds to C1q in a binding stoichiometric ratio in the range of 20:1 to 1.0:1 or less than 1.0:1. In certain embodiments, the anti-C1q antibody binds to C1q in a binding stoichiometric ratio in the range of 6:1 to 1.0:1 or less than 1.0:1. In certain embodiments, the anti-C1q antibody binds to C1q in a binding stoichiometry ranging from 2.5:1 to 1.0:1 or less than 1.0:1. In some embodiments, the anti-C1q antibody inhibits the interaction between C1q and C1r, or between C1q and C1s, or between C1q and both C1r and C1s.In some embodiments, the anti-C1q antibody inhibits the interaction between C1q and C1r, between C1q and C1s, and / or between C1q and both C1r and C1s. In some embodiments, the anti-C1q antibody binds to the A chain of C1q. In other embodiments, the anti-C1q antibody binds to the B chain of C1q. In other embodiments, the anti-C1q antibody binds to the C chain of C1q. In some embodiments, the anti-C1q antibody binds to the A chain, B chain, and / or C chain of C1q. In some embodiments, the anti-C1q antibody binds to the globular domains of the A chain, B chain, and / or C chain of C1q. In other embodiments, the anti-C1q antibody binds to the collagen-like domains of the A chain, B chain, and / or C chain of C1q.

[0127] If the antibody of this disclosure inhibits the interaction between two or more complement factors, for example, the interaction between C1q and C1s, or the interaction between C1q and C1r, the interaction occurring in the presence of the antibody may be reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% compared to a control in the absence of the antibody of this disclosure. In some embodiments, the antibody of this disclosure reduces the interaction between two or more complement factors by 50%, 60%, 70%, 80%, 90%, or 100%. In certain embodiments, the interaction occurring in the presence of the antibody is reduced by an amount ranging from at least 30% to at least 99% compared to a control in the absence of the antibody of this disclosure.

[0128] In some embodiments, the antibodies of the Disclosure inhibit C2 or C4 cleavage by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%, or in amounts ranging from at least 30% to at least 99%, compared to a control in which the antibodies of the Disclosure are absent. Methods for measuring C2 or C4 cleavage are well known in the art. EC for the antibodies of the Disclosure at each C2 or C4 cleavage.50 The values ​​may be less than 3 μg / ml; 2.5 μg / ml; 2.0 μg / ml; 1.5 μg / ml; 1.0 μg / ml; 0.5 μg / ml; 0.25 μg / ml; 0.1 μg / ml; or 0.05 μg / ml. In some embodiments, the antibodies of this disclosure inhibit C2 or C4 cleavage with approximately equimolar concentrations of C1q and their respective anti-C1q antibodies.

[0129] In some embodiments, the antibodies of the Disclosure inhibit autoantibody-dependent and complement-dependent cell-mediated cytotoxicity (CDC) by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%, or in amounts ranging from at least 30% to at least 99%, compared to a control in which the antibodies of the Disclosure are absent. 50 The values ​​may be less than 3 μg / ml; 2.5 μg / ml; 2.0 μg / ml; 1.5 μg / ml; 1.0 μg / ml; 0.5 μg / ml; 0.25 μg / ml; 0.1 μg / ml; or 0.05 μg / ml.

[0130] In some embodiments, the antibodies of the Disclosure inhibit complement-dependent cell-mediated cytotoxicity (CDCC) by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%, or in amounts ranging from at least 30% to at least 99%, compared to a control in which the antibodies of the Disclosure are absent. Methods for measuring CDCC are well known in the art. Regarding CDCC inhibition, the EC of the antibodies of the Disclosure 50 The values ​​may be less than 3 μg / ml; 2.5 μg / ml; 2.0 μg / ml; 1.5 μg / ml; 1.0 μg / ml; 0.5 μg / ml; 0.25 μg / ml; 0.1 μg / ml; or 0.05 μg / ml. In some embodiments, the antibodies of this disclosure inhibit CDCC but do not inhibit antibody-dependent cell-mediated cytotoxicity (ADCC).

[0131] Humanized anti-complement C1q antibody The humanized antibodies of this disclosure specifically bind to complement factor C1q and / or C1q protein in the C1 complex of the classical complement pathway. Humanized anti-C1q antibodies may specifically bind to human C1q, human and mouse C1q, rat C1q, or human C1q, mouse C1q, and rat C1q.

[0132] All sequences cited in the following 16 paragraphs are incorporated herein by reference from U.S. Patent Application No. 14 / 933,517 (which is incorporated herein by reference with respect to the antibodies and related compositions disclosed therein).

[0133] In some embodiments, the human heavy chain constant region is a human IgG4 heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 47, or an amino acid sequence having at least 70%, at least about 75%, at least about 80%, at least about 85%, or at least about 90% homology to SEQ ID NO: 47. The human IgG4 heavy chain constant region may comprise an Fc region having one or more modifications and / or amino acid substitutions according to Kabat numbering. In such cases, the Fc region comprises an amino acid substitution from leucine to glutamate at position 248, such substitution inhibiting the Fc region from interacting with the Fc receptor. In some embodiments, the Fc region comprises an amino acid substitution from serine to proline at position 241, such substitution preventing arm switching in the antibody.

[0134] The amino acid sequence of the human IgG4 (S241P L248E) heavy chain constant domain is ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (Sequence ID 47).

[0135] The antibody may include a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprising an amino acid sequence selected from any one of SEQ ID NOs: 31-34, or an amino acid sequence having at least about 90% homology to an amino acid sequence selected from any one of SEQ ID NOs: 31-34. In certain such embodiments, the light chain variable domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 35-38, or an amino acid sequence having at least about 90% homology to an amino acid sequence selected from any one of SEQ ID NOs: 35-38.

[0136] The amino acid sequence of heavy chain variable domain variant 1 (VH1) is:

[0137] The filename is TIFF2023551734000005.tif25170. The hypervariable region (HVR) of VH1 is indicated in bold and underlined text.

[0138] The amino acid sequence of heavy chain variable domain variant 2 (VH2) is:

[0139] The filename is TIFF2023551734000006.tif25170. The hypervariable region (HVR) of VH2 is shown in bold and underlined text.

[0140] The amino acid sequence of heavy chain variable domain variant 3 (VH3) is:

[0141] The filename is TIFF2023551734000007.tif25170. The VH3 hypervariable region (HVR) is indicated in bold and underlined text.

[0142] The amino acid sequence of heavy chain variable domain variant 4 (VH4) is:

[0143] The filename is TIFF2023551734000008.tif25170. The VH4 hypervariable region (HVR) is indicated in bold and underlined text.

[0144] The amino acid sequence of kappa light chain variable domain variant 1 (Vκ1) is:

[0145] The file is TIFF2023551734000009.tif19170. The hypervariable region (HVR) of Vκ1 is shown in bold and underlined text.

[0146] The amino acid sequence of kappa light chain variable domain variant 2 (Vκ2) is:

[0147] The file is TIFF2023551734000010.tif19170. The hypervariable region (HVR) of Vκ2 is shown in bold and underlined text.

[0148] The amino acid sequence of kappa light chain variable domain variant 3 (Vκ3) is:

[0149] The file is TIFF2023551734000011.tif19170. The hypervariable region (HVR) of Vκ3 is shown in bold and underlined text.

[0150] The amino acid sequence of kappa light chain variable domain variant 4 (Vκ4) is:

[0151] The image is TIFF2023551734000012.tif19170. The hypervariable region (HVR) of Vκ4 is shown in bold and underlined text.

[0152] The antibody may contain at least 85%, 90%, or 95% identical light chain variable domain amino acid sequences to sequences 35-38, while retaining HVR-L1 RASKSINKYLA (SEQ ID NO: 5), HVR-L2 SGSTLQS (SEQ ID NO: 6), and HVR-L3 QQHNEYPLT (SEQ ID NO: 7). Preferably, the antibody may contain at least 85%, 90%, or 95% identical heavy chain variable domain amino acid sequences to sequences 31-34, while retaining HVR-H1 GYHFTSYWMH (SEQ ID NO: 9), HVR-H2 VIHPNSGSINYNEKFES (SEQ ID NO: 10), and HVR-H3 ERDSTEVLPMDY (SEQ ID NO: 11).

[0153] In some embodiments, the antibody includes the light chain variable domain amino acid sequence of SEQ ID NO: 35 and the heavy chain variable domain amino acid sequence of SEQ ID NO: 31. In some embodiments, the antibody includes the light chain variable domain amino acid sequence of SEQ ID NO: 36 and the heavy chain variable domain amino acid sequence of SEQ ID NO: 32. In some embodiments, the antibody includes the light chain variable domain amino acid sequence of SEQ ID NO: 37 and the heavy chain variable domain amino acid sequence of SEQ ID NO: 33. In some embodiments, the antibody includes the light chain variable domain amino acid sequence of SEQ ID NO: 38 and the heavy chain variable domain amino acid sequence of SEQ ID NO: 34.

[0154] In some embodiments, the humanized anti-C1q antibody of the Disclosure comprises a heavy chain variable region containing a Fab region and a heavy chain constant region containing an Fc region, wherein the Fab region specifically binds to the C1q protein of the Disclosure, but the Fc region is unable to bind to the C1q protein. In some embodiments, the Fc region is derived from a human IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the Fc region is unable to induce complement activity and / or antibody-dependent cell-mediated cytotoxicity (ADCC). In some embodiments, the Fc region includes one or more modifications, including, but not limited to, amino acid substitutions. In certain embodiments, the Fc region of the humanized anti-C1q antibody of the Disclosure includes an amino acid substitution at position 248 or a position corresponding to position 248 according to the Kabat numbering convention, and / or at position 241 or a position corresponding to position 241 according to the Kabat numbering convention. In some embodiments, an amino acid substitution at position 248 or a position corresponding to position 248 inhibits the Fc region from interacting with the Fc receptor. In some embodiments, the amino acid substitution at position 248 or a position corresponding to position 248 is an amino acid substitution from leucine to glutamate. In some embodiments, an amino acid substitution at position 241 or a position corresponding to position 241 prevents arm switching in the antibody. In some embodiments, the amino acid substitution at position 241 or a position corresponding to position 241 is an amino acid substitution from serine to proline. In certain embodiments, the Fc region of the humanized anti-C1q antibody of the present disclosure includes the amino acid sequence of SEQ ID NO: 47, or an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% homology to the amino acid sequence of SEQ ID NO: 47.

[0155] Anti-C1q Fab fragment Before the advent of recombinant DNA technology, proteases, which cleave polypeptide sequences, were used to cleave the structure of antibody molecules and determine which parts of the molecule are responsible for their various functions. Limited digestion with the protease papain cleaves the antibody molecule into three fragments. Two of these fragments, known as Fab fragments, are identical and contain antigen-binding activity. The Fab fragments correspond to two identical arms of the antibody molecule, each of which is the V of the heavy chain. H and C H It consists of a complete light chain paired with one domain. Other fragments, while lacking antigen-binding activity, were initially observed to crystallize readily and were therefore named Fc fragments (crystallizable fragments). When Fab molecules were compared to IgG molecules, Fab was found to be superior to IgG for certain in vivo applications due to their higher mobility and tissue penetration ability, their reduced circulating half-life, their ability to bind to antigens monovalently without mediating antibody effector function, and their lower immunogenicity.

[0156] The Fab molecule has a constant domain C H 2 and C H It is an artificial fragment of an Ig molecule with a heavy chain shortened by 3, approximately 50 kDa. L -V H and C L -C H 1) Domain interaction is fundamental to the structure of the two chains of the Fab molecule, and this is C L and C H Further stabilization is achieved by disulfide crosslinking with 1. Fab and IgG have six complementarity-determining regions (CDRs) (three of which are V L and V H They have the same antigen-binding site formed by (LCDR1, LCDR2, LCDR3 and HCDR1, HCDR2, HCDR3) derived from the antibody. CDRs define the hypervariable antigen-binding site of the antibody. The highest sequence variation is found in LCDR3 and HCDR3, which are V in the innate immune system, respectively. L and J L gene or V H ,D H and J It is generated by rearrangement of the H gene. LCDR3 and HCDR3 typically form the core of the antigen-binding site. The conserved region that connects and presents the six CDRs is called the framework region. In the three-dimensional structure of the variable domain, the framework region forms a sandwich of two opposing antiparallel β-sheets linked by hypervariable CDR loops on the outside and conserved disulfide crosslinks on the inside. This unique combination of the stability and versatility of the antigen-binding site of Fab and IgG highlights its success in clinical work for disease diagnosis, monitoring, prevention, and treatment.

[0157] All anti-C1q antibody Fab fragment sequences are incorporated herein by reference from U.S. Patent Application No. 15 / 360,549 (which is incorporated herein by reference with respect to the antibodies and associated compositions disclosed therein).

[0158] In certain embodiments, the present disclosure relates to heavy chain (V H / C H 1) and light chain (V L / C L An anti-C1q antibody Fab fragment that binds to the C1q protein containing ), the anti-C1q antibody Fab fragment has six complementation-determining regions (CDRs) (three of which are V L and V H The present invention provides an anti-C1q antibody Fab fragment having (HCDR1, HCDR2, HCDR3, and LCDR1, LCDR2, LCDR3) derived from IgG1. The heavy chain of the antibody Fab fragment is cleaved after the first heavy chain domain of IgG1 (SEQ ID NO: 39) and contains the following amino acid sequence:

[0159] The complementarity determination region (CDR) of sequence number 39 in TIFF2023551734000013.tif37170 is shown in bold and underlined text.

[0160] The light chain domain of the antibody Fab fragment contains the following amino acid sequence (SEQ ID NO: 40):

[0161] In TIFF2023551734000014.tif, the complementarity-determining region (CDR) of SEQ ID NO: 40 is indicated by bold and underlined characters.

[0162] Nucleic Acids, Vectors, and Host Cells Antibodies suitable for use in the methods of the present disclosure can be produced using recombinant methods and compositions, for example, as described in U.S. Patent No. 4,816,567. In some embodiments, an isolated nucleic acid having a nucleotide sequence encoding any of the antibodies of the present disclosure is provided. Such nucleic acids encode the amino acid sequence containing the V L / C L and / or the amino acid sequence containing the V H / C H 1. In some embodiments, one or more vectors (e.g., expression vectors) containing such nucleic acids are provided. Host cells containing such nucleic acids can also be provided. The host cell contains (1) a vector containing a nucleic acid encoding the amino acid sequence containing the V L / C L of the antibody and the amino acid sequence containing the V H / C H 1 of the antibody, or (2) a first vector containing a nucleic acid encoding the amino acid sequence containing the V L / C L of the antibody and a second vector containing a nucleic acid encoding the amino acid sequence containing the V H / C H 1 of the antibody (e.g., being transfected). In some embodiments, the host cell is eukaryotic, for example, Chinese hamster ovary (CHO) cells or lymphocytes (e.g., Y0, NS0, Sp20 cells). In some embodiments, the host cell is a bacterium such as E. coli.

[0163] Methods for making anti-C1q antibodies are disclosed herein. The method includes culturing a host cell of the present disclosure containing a nucleic acid encoding an anti-C1q antibody under conditions suitable for the expression of the antibody. In some embodiments, the antibody is then recovered from the host cell (or the host cell culture medium).

[0164] For the recombinant generation of the humanized anti-C1q antibody of this disclosure, the nucleic acid encoding the antibody is isolated and inserted into one or more vectors for further cloning and / or expression in host cells. Such nucleic acids can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of specifically binding to the genes encoding the heavy and light chains of the antibody).

[0165] Suitable vectors containing nucleic acid sequences encoding any of the antibodies of this disclosure or their fragment polypeptides (including antibodies) described herein include, but are not limited to, cloning vectors and expression vectors. Suitable cloning vectors may be constructed according to standard techniques or selected from a large number of cloning vectors available in the art. The cloning vector selected may vary depending on the host cell to which it is intended to be used, while useful cloning vectors typically have the ability to self-replicate, may possess a single target for a particular restriction endonuclease, and / or may have genes for markers that may be used when selecting a clone containing the vector. Suitable examples include plasmids and bacterial viruses, e.g., pUC18, pUC19, Bluescript (e.g., pBS SK+) and its derivatives, mpl8, mpl9, pBR322, pMB9, ColE1, pCR1, RP4, phage DNA, and shuttle vectors, e.g., pSA3 and pAT28. These and many other cloning vectors are available from commercial vendors such as BioRad, Stratagene, and Invitrogen.

[0166] Vectors containing the target nucleic acid can be introduced into host cells by any of a number of suitable means, including electroporation, transfection using calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, or other substances; particulate guns; lipofection; and infection (e.g., the vector is an infectious agent such as vaccinia virus). The choice of introducing a vector or polynucleotide often depends on the characteristics of the host cell. In some embodiments, the vector contains nucleic acid comprising one or more amino acid sequences encoding the anti-C1q antibody of this disclosure.

[0167] Suitable host cells for cloning or expressing antibody-encoding vectors include prokaryotic or eukaryotic cells. For example, the anti-C1q antibody of this disclosure may be generated in bacteria, particularly when glycosylation and Fc effector function are not required. Regarding the expression of antibody fragments and polypeptides in bacteria (e.g., U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523; and Charlton, Methods in Molecular Biology, Vol. 248 (BKCLo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254 (described regarding the expression of antibody fragments in E. coli)). In other embodiments, the antibodies of this disclosure may be generated in eukaryotic cells, such as Chinese hamster ovary (CHO) cells or lymphocytes (e.g., Y0, NS0, Sp20 cells) (e.g., U.S. Patent Application No. 14 / 269,950, U.S. Patent No. 8,981,071, Eur J Biochem. 1991 Jan 1;195(1):235-42). After expression, the antibodies may be isolated from bacterial cell paste in a soluble fraction and further purified.

[0168] Pharmaceutical composition and administration The anti-C1q antibody of this disclosure (e.g., FabA) may be administered in the form of a pharmaceutical composition.

[0169] Therapeutic formulations of antibodies, antibody fragments and / or antibody derivatives of this disclosure may be prepared in the form of lyophilized formulations or aqueous solutions for storage by mixing antibodies of desired purity with any pharmaceutically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed.

[1980] ). Acceptable carriers, excipients or stabilizers are nontoxic to the recipient at the dosage and concentration used and include buffers, e.g., phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkylparabens, e.g., methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, e.g., These include serum albumin, gelatin, or immunoglobulin; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, e.g., EDTA; sugars, e.g., sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, e.g., sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, e.g., TWEEN®, PLURONICS®, or polyethylene glycol (PEG).

[0170] Lipofection or liposomes can also be used to deliver antibodies or antibody fragments, or antibody derivatives, to cells, preferably epitopes or minimal fragments that specifically bind to the binding domain of the target protein.

[0171] Antibodies can also be incorporated into colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsifies, nanoparticles, and nanocapsules) or macroemulsifies, for example, within microcapsules prepared by coacervation technology or interfacial polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly-(methyl methacrylate) microcapsules, respectively. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).

[0172] The formulation used for administration may be sterile. This can be easily achieved by filtration through a sterile filtration membrane.

[0173] Sustained-release preparations can be prepared. Preferred examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing antibodies, which may be in the form of molded articles, e.g., films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactide (U.S. Patent No. 3,773,919), copolymers of L-glutamic acid and γ-ethyl-L-glutamic acid ester, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers, e.g., LUPRON DEPOT® (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyrate. Polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid allow for molecular release over 100 days, while certain hydrogels release proteins over shorter periods.

[0174] The antibodies, antibody fragments and / or antibody derivatives and compositions of this disclosure are typically administered by intravitreous administration.

[0175] A pharmaceutical composition may also include a pharmaceutically acceptable, non-toxic carrier of a diluent, defined as a vehicle commonly used to formulate the pharmaceutical composition for animal or human administration, depending on the desired formulation. The diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents include distilled water, buffered water, physiological saline, PBS, Ringer's solution, dextrose solution, and Hanks' solution. The pharmaceutical composition or formulation may also include other carriers, adjuvants, or non-toxic, non-therapeutic, non-immunogenic stabilizers, excipients, etc. The composition may also include additional substances for approximate physiological conditions, such as pH adjusters and buffers, toxicity modifiers, wetting agents, and detergents.

[0176] The composition may also contain, for example, one of a variety of stabilizers, such as antioxidants. If the pharmaceutical composition contains a polypeptide, the polypeptide may be complexed with a variety of well-known compounds that improve the polypeptide's in vivo stability or otherwise enhance its pharmacological properties (e.g., increasing the polypeptide's half-life, reducing its toxicity, improving other pharmacokinetic and / or pharmacodynamic properties, or improving solubility or uptake). Examples of such modifiers or complexing agents include sulfates, glucons, citrates, and phosphates. Polypeptides in a composition may also be complexed with molecules that improve their in vivo properties. Such molecules include, for example, carbohydrates, polyamines, amino acids, other peptides, ions (e.g., sodium, potassium, calcium, magnesium, manganese), and lipids. Further guidance on formulations suitable for various types of administration can be found in Remington's Pharmaceutical Sciences, Mace Publishing Company, Philadelphia, Pa., 17th ed. (1985). For a brief review of drug delivery methods, see Langer, Science 249:1527-1533 (1990).

[0177] The toxicity and therapeutic efficacy of the active ingredient can be determined by standard pharmaceutical procedures in cell culture and / or experimental animals, for example, by determining the LD50 (lethal dose for 50% of the population) and ED50 (therapeutably effective dose for 50% of the population). The dose ratio between toxicity and therapeutic effect is the therapeutic index, which can be expressed as the ratio LD50 / ED50. Compounds exhibiting a high therapeutic index are preferred.

[0178] Data obtained from cell cultures and / or animal studies and / or human clinical trials may be used in formulating dosage ranges for humans. Doses of the active ingredient typically fall within the range of circulating concentrations, including ED50, with low toxicity. Doses may vary within this range depending on the form of administration and the route of administration used.

[0179] The components used to formulate pharmaceutical compositions are preferably of high purity and substantially free of potentially harmful contaminants (e.g., at least National Food grade, usually at least analytical grade, more typically at least pharmaceutical grade). Furthermore, compositions intended for parenteral use are usually sterile. As long as a given compound must be synthesized before use, the resulting product is typically substantially free of any potentially toxic substances, in particular any endotoxins, that may be present during the synthesis or purification process. Compositions for parenteral administration are also typically substantially isotonic and produced under GMP conditions.

[0180] The compositions of this disclosure may be administered by any medically appropriate procedure, for example, intravitreal injection.

[0181] Treatment method This disclosure generally relates to compositions and methods for preventing, reducing the risk of developing, or treating eye diseases in human patients, such as glaucoma or age-related macular degeneration, including AMD with geographic atrophy. Such methods involve administering to a patient, via intravitreal injection, a composition containing about 1 mg to about 10 mg of anti-C1q antibody (e.g., about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, or about 10 mg of anti-C1q antibody). Such a method also involves administering to a patient, via intravitreal injection, a composition comprising an anti-C1q antibody in an amount of approximately 1 mg to approximately 10 mg (for example, approximately 1 mg, approximately 1.5 mg, approximately 2 mg, approximately 2.5 mg, approximately 3 mg, approximately 3.5 mg, approximately 4 mg, approximately 4.5 mg, approximately 5 mg, approximately 5.5 mg, approximately 6 mg, approximately 6.5 mg, approximately 7 mg, approximately 7.5 mg, approximately 8 mg, approximately 8.5 mg, approximately 9 mg, approximately 9.5 mg, or approximately 10 mg of anti-C1q antibody), wherein the antibody comprises a light chain variable domain comprising HVR-L1 having the amino acid sequence of SEQ ID NO: 5, HVR-L2 having the amino acid sequence of SEQ ID NO: 6, and HVR-L3 having the amino acid sequence of SEQ ID NO: 7, and a heavy chain variable domain comprising HVR-H1 having the amino acid sequence of SEQ ID NO: 9, HVR-H2 having the amino acid sequence of SEQ ID NO: 10, and HVR-H3 having the amino acid sequence of SEQ ID NO: 11. The administered composition may contain about 1 mg to about 5 mg of anti-C1q antibody. The administered composition may contain about 1 mg to about 2.5 mg, about 2.5 mg to about 5 mg, about 5 mg to about 7.5 mg, or about 7.5 mg to about 10 mg of anti-C1q antibody. The administered composition may contain about 5 mg of anti-C1q antibody. The administered composition may contain about 10 mg of anti-C1q antibody. In some embodiments, the antibody includes a light chain variable domain having an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs. 4 and 35-38, wherein the light chain variable domain includes HVR-L1 having the amino acid sequence of SEQ ID NO. 5, HVR-L2 having the amino acid sequence of SEQ ID NO. 6, and HVR-L3 having the amino acid sequence of SEQ ID NO. 7.In some embodiments, the light chain variable domain includes an amino acid sequence selected from SEQ ID NOs: 4 and 35-38. In some embodiments, the antibody includes a heavy chain variable domain containing an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs: 8 and 31-34, wherein the heavy chain variable domain includes HVR-H1 having the amino acid sequence of SEQ ID NOs: 9, HVR-H2 having the amino acid sequence of SEQ ID NOs: 10, and HVR-H3 having the amino acid sequence of SEQ ID NOs: 11. In some embodiments, the heavy chain variable domain includes an amino acid sequence selected from SEQ ID NOs: 8 and 31-34. In some embodiments, the antibody comprises a light chain variable domain having an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs: 4 and 35-38, comprising HVR-L1 having the amino acid sequence of SEQ ID NO: 5, HVR-L2 having the amino acid sequence of SEQ ID NO: 6, and HVR-L3 having the amino acid sequence of SEQ ID NO: 7; and a heavy chain variable domain having an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs: 8 and 31-34, comprising HVR-H1 having the amino acid sequence of SEQ ID NO: 9, HVR-H2 having the amino acid sequence of SEQ ID NO: 10, and HVR-H3 having the amino acid sequence of SEQ ID NO: 11. In some embodiments, the antibody comprises a light chain variable domain having an amino acid sequence selected from SEQ ID NOs: 4 and 35-38, and a heavy chain variable domain having an amino acid sequence selected from SEQ ID NOs: 8 and 31-34. In some embodiments, the antibody may be a monoclonal antibody, a humanized antibody, a human antibody, a chimeric antibody, an antibody fragment, or an antibody derivative thereof. The antibody fragment may be a Fab fragment, a Fab' fragment, an F(ab')2 fragment, an Fv fragment, a diabody, or a single-chain antibody molecule. In some embodiments, the Fab fragment includes the heavy chain Fab fragment of SEQ ID NO: 39 and the light chain Fab fragment of SEQ ID NO: 40.

[0182] In some embodiments, the antibody is administered once a week, once every two weeks, once every three weeks, once a month, once every four weeks, once every six weeks, once every eight weeks, once every two months, once every ten weeks, once every twelve weeks, once every three months, or once every four months. In some embodiments, the antibody is administered for at least six months, at least seven months, at least eight months, at least nine months, at least ten months, at least eleven months, or at least twelve months.

[0183] In some embodiments, the eye disease is glaucoma or age-related macular degeneration such as geographic atrophy.

[0184] In certain preferred embodiments, FabA is administered as an IVT injection at a dose of 2.5 mg / eye, once a month, once every four weeks, once every six weeks, or once every two months.

[0185] In certain preferred embodiments, FabA is administered as an IVT injection at a dose of 5 mg / eye once a month, once every four weeks, once every six weeks, or every other month.

[0186] In certain preferred embodiments, FabA is administered as an IVT injection at a dose of 5 mg / eye, once a month, once every four weeks, once every six weeks, or once every two months.

[0187] In certain preferred embodiments, FabA is administered as an IVT injection at a dose of 10 mg / eye, once a month, once every four weeks, once every six weeks, or once every two months.

[0188] FabA injections are administered by trained and qualified physicians using sterile techniques.

[0189] Anti-C1q antibodies may inhibit interactions between C1q and autoantibodies, between C1q and C1r, or between C1q and C1s, or may promote the clearance of C1q from circulation or tissue. In some embodiments, the anti-C1q antibody has a dissociation constant (K) in the range of 100 nM to 0.005 nM or less than 0.005 nM. D ) has. In some embodiments, the anti-C1q antibody binds to C1q in a binding stoichiometry of 20:1 to 1.0:1 or less than 1.0:1, 6:1 to 1.0:1 or less than 1.0:1, or 2.5:1 to 1.0:1 or less than 1.0:1.

[0190] This method inhibits the biological activity of C1q. For example, (1) C1q binding to autoantibodies, (2) C1q binding to C1r, (3) C1q binding to C1s, (4) C1q binding to phosphatidylserine, (5) C1q binding to pentraxin-3, (6) C1q binding to C-reactive protein (CRP), (7) C1q binding to globular C1q receptor (gC1qR), (8) C1q binding to complement receptor 1 (CR1), (9) C1q binding to B amyloid, or (10) C1q binding to calreticulin.In other embodiments, the biological activity of C1q includes: (1) activation of the classical complement activation pathway, (2) decreased lysis and / or decreased C3 deposition, (3) activation of antibody and complement-dependent cell cytotoxicity, (4) CH50 hemolysis, (5) decreased erythrocyte lysis, (6) decreased erythrocyte phagocytosis, (7) decreased dendritic cell infiltration, (8) inhibition of complement-mediated erythrocyte lysis, (9) decreased lymphocyte infiltration, (10) decreased macrophage infiltration, (11) decreased antibody deposition, and (12) (13) Decreased neutrophil infiltration, (14) Decreased platelet phagocytosis, (15) Improved graft survival, (16) Decreased macrophage-mediated phagocytosis, (17) Decreased autoantibody-mediated complement activation, (18) Decreased erythrocyte destruction due to transfusion, (19) Decreased erythrocyte lysis due to alloantibodies, (20) Decreased hemolysis due to transfusion, (21) Decreased alloantibody-mediated platelet lysis, (22) Improved anemia, (23) Decreased eosinophilia, (24 (25) Decreased C3 deposition on red blood cells (e.g., decreased deposition of C3b, iC3b, etc. on RBCs), (26) Decreased C3 deposition on platelets (e.g., decreased deposition of C3b, iC3b, etc. on platelets), (27) Decreased anaphylatoxin production, (28) Decreased autoantibody-mediated rash formation, (29) Decreased autoantibody-induced lupus erythematosus, (30) Decreased red blood cell destruction due to transfusion reaction, (31) Decreased platelet lysis due to transfusion reaction, (32) Obesity (32) decreased cell activation, (33) decreased mast cell histamine release, (34) decreased vascular permeability, (35) decreased complement deposition on graft endothelium, (36) B cell antibody production, (37) dendritic cell maturation, (38) T cell proliferation, (39) cytokine production, (40) microglia activation, (41) decreased Artus reaction, (42) decreased anaphylatoxin production in graft endothelium, or (42) activation of complement receptor 3 (CR3 / C3) expressing cells.

[0191] In some embodiments, CH50 hemolysis includes human CH50 hemolysis. The antibody may be capable of neutralizing at least about 50% to about 100% of human CH50 hemolysis. The antibody may be capable of neutralizing about 50%, about 60%, about 70%, about 80%, about 90%, and about 100% of human CH50 hemolysis. The antibody may be capable of neutralizing at least 50% of CH50 hemolysis at doses of less than 150 ng / ml, less than 100 ng / ml, less than 50 ng / ml, or less than 20 ng / ml.

[0192] In some embodiments, the antibody is a monoclonal antibody, polyclonal antibody, recombinant antibody, humanized antibody, human antibody, chimeric antibody, monovalent antibody, multispecific antibody, or antibody fragment, or an antibody derivative thereof. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is an antibody fragment such as a Fab fragment. Examples of antibody fragments are Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, diabodies, and single-chain antibody molecules.

[0193] The compositions are intended to be obtained and used under the guidance of a physician for in vivo use. The dosage of the therapeutic formulation may vary widely depending on the characteristics of the disease, the frequency of administration, the mode of administration, and the clearance of the drug from the host.

[0194] As used herein, “chronically administered,” “chronically treated,” “to treat chronically,” or similar grammatical types refer to a treatment regimen used to maintain a predetermined threshold concentration of a therapeutic agent in a patient’s eye in order to completely or substantially suppress systemic complement activity in the patient over a long period of time. Thus, a patient chronically treated with an antibody is a patient who has been treated for two weeks or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, or 52 weeks; 1, 2, 3, The patient may be treated with an amount and frequency of antibody sufficient to maintain an antibody concentration in the patient's eye that inhibits or substantially inhibits systemic complement activity in the patient for a period of 4, 5, 6, 7, 8, 9, 10, 11, or 12 months; or 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, or 12 years or for the remainder of the patient's life. In some embodiments, the antibody may be administered chronically to patients who require it in an amount and frequency effective to maintain serum hemolytic activity at 20% or less (e.g., 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or even less than 5%). In some embodiments, the antibody may be administered to the patient in an amount and frequency effective in maintaining serum lactate dehydrogenase (LDH) levels within at least 20% of the normal range for LDH (e.g., 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or even less than 5%).

[0195] Therapeutic agents, such as anti-C1q antibodies, can be incorporated into various formulations for therapeutic administration by combining them with appropriate, pharmaceutically acceptable carriers or diluents. [Examples]

[0196] Example 1: Evaluation of FabA in non-clinical trials FabA is a sterile isotonic solution for in vitro venous therapy (IVT) injection.

[0197] FabA is supplied as a sterile, single-dose vial for IVT injection.

[0198] A wide range of in vitro and in vivo pharmacological studies were conducted using FabA.

[0199] The antibodies Mab1, Mab1-Fab, and Mab2 were active in an acute glaucoma mouse model and protected retinal ganglion cells and / or nerve fibers from loss. In a photo-oxidation-induced injury model mouse, intravitreal administration of Mab1 had a protective effect against intraocular photoreceptor cell loss and retinal functional connectivity.

[0200] The GLP studies for FabA consisted of a single-dose rat ocular toxicity study and a three-dose cynomolgus monkey ocular toxicity study. The route of administration for the toxicity studies was IVT injection. In the single-dose and twice-dose (once a month) IVT GLP studies, FabA did not show evidence of adverse ocular toxicity at the no-observed-adverse-effect level (NOAEL) of 5 mg / eye (equivalent to a human dose of 10 mg) administered once a month in cynomolgus monkeys, and 0.05 mg / eye (equivalent to a human dose of 10 mg) in the single-dose rat study. In the 26-week chronic ocular toxicity study in cynomolgus monkeys, adverse ocular changes were associated with the two injection procedures and / or were determined to be mediated by anti-drug antibodies (ADAs) and not a direct effect of FabA IVT administration.

[0201] Pharmacokinetic evaluations of FabA were performed in the serum and vitreous humor of rats and cynomolgus monkeys. Intravitreous C1q concentration was measured as a PD marker for FabA-mediated C1q inhibition in monkeys. PK / PD and TK / PD studies in monkeys demonstrated a robust ocular PD effect consistent with FabA drug exposure levels in the vitreous humor.

[0202] Binding and affinity of FabA and precursor molecules to human C1q. The binding affinity of FabA and its precursor molecules to C1q was also investigated by ELISA. All molecules (Mab2-Fab, FabA, and Mab2, Mab1, and Mab1-Fab) showed affinity for human C1q and had 50% effective concentrations (EC50) in the range of 2.2–4.9 ng / mL (20–95 pM). The EC50 for FabA binding to C1q was 2.5 ng / mL.

[0203] Effects on IgM-mediated erythrolysis The activities of FabA, Mab2, and Mab2-Fab, which functionally inhibit classical complement-dependent hemolysis of IgM-opsonized RBCs in human serum, were measured (Figure 3). The three molecules showed almost identical titers, which is consistent with equivalent binding affinities. The 50% inhibitory concentration (IC50) of FabA inhibition against IgM-coated RBC hemolysis was 0.62 μg / mL (approximately 12 nm).

[0204] In vivo pharmacological examination Prevention of optic nerve damage by anti-C1q antibody treatment in an acute glaucoma mouse model. In mice, injection of polystyrene beads into the anterior chamber induced a rapid increase in IOP, retinal ganglion cell loss, and optic nerve damage over a two-week period. Mab1, Mab1-Fab, and Mab2 were administered intravitreally to the mice the day before and 7 days after the IOP increase. At each time point, 2 μL of 10 mg / mL antibody or saline was administered. Based on a vitreous volume of 5–10 μL in the mouse eye, antibody concentrations ranged from 2000–4000 μg / mL. Optic nerves were collected two weeks after injury, and the number of intact and damaged axons was quantified. Anti-C1q antibody treatment provided protection against RGC loss and / or retinal nerve fiber damage in this glaucoma-induced mouse model (Figure 4).

[0205] Protection from photoreceptor cell damage by anti-C1q antibody treatment in a photooxidation-induced injury model. When mice were exposed to 100 Klux of natural white LED light for 1–7 days, photooxidative damage resulted in retinal photoreceptor cell loss. In this model, there was a time-dependent increase in C1qa gene expression over 3–7 days, correlated with photoreceptor cell death and microglia / macrophage recruitment. C1qa- / - mice did not exhibit these characteristics 7 days after photoinjury induction, but at 14 days, they showed less photoreceptor cell death, decreased microglia / macrophage recruitment to photoreceptor cell damage, and higher visual function. In vitro administration of Mab1 antibody on day 7 after photoinjury reduced photoreceptor cell loss and maintained retinal function, as measured by electroretinography (Figure 5). Mice were administered 1 μL of 7.5 mg / mL antibody, which corresponds to intravitreal concentrations of 750–1500 ug / mL. In contrast, systemic delivery of 100 mg / kg of Mab1 on days 0, 4, and 8 had no effect on photoreceptor cell loss or function. Retinal C1q was primarily expressed in the outer retina in early AMD and by subretinal microglia / macrophages located in the mouse retina. Therefore, protection with anti-C1q antibodies suggests a clear role of C1q in the pathogenesis of GA in human disease, specifically in photoreceptor damage and the initiation of the classical complement cascade.

[0206] Safety pharmacology In a 26-week study of cynomolgus monkeys, systemic exposure following chronic in vitro thromboembolism (IVT) administration did not exceed 86.3 ng / mL. In contrast, in a 26-week study of cynomolgus monkeys, systemic exposure to Mab2 with the same CDR (Chemical Distributed Ratio) from weekly IV administration exceeded 1 mg / mL at a NOAEL (No Observed Adverse Emission Level) of 200 mg / kg.

[0207] Therefore, the safety pharmacological endpoints of the full-length antibody, Mab2, after IV administration were up to 200 mg / kg per week in a 4-week repeated-dose GLP toxicity study in monkeys and up to 200 mg / kg per week in a 26-week repeated-dose toxicity study in monkeys. There was no evidence of treatment-related effects on cardiovascular, respiratory, or nervous system endpoints, which supported the systemic safety of FabA administered via IVT.

[0208] Furthermore, the safety pharmacological endpoint of FabA after SC administration was a maximum of 20 mg / kg daily in a 4-week repeated-dose GLP toxicity study in monkeys, with no evidence of treatment-related effects on cardiovascular, respiratory, or nervous system endpoints, thus supporting the systemic safety of FabA administered via IVT.

[0209] Pharmacokinetics in Animals Non-clinical studies designed to characterize the pharmacokinetic (PK), toxin-keratin-zoster (TK), and pharmacokinetic (PD) effects of FabA were conducted in rats and cynomolgus monkeys. These studies included single-dose IVT PK studies and repeated-dose TK / PD studies in cynomolgus monkeys using FabA. Larger TK / PD studies were conducted in monkeys, and no ocular toxicity was observed in either the rat or monkey single-dose studies.

[0210] Pharmacokinetics / Toxicology / Pharmacodynamic Analysis Pharmacokinetics of FabA in the Vitreous Intravitreous In rats, FabA was administered as a single dose to both in vitro vitreous tracts (IVTs) at doses of 0.01 mg / eye (equivalent to a human dose of 2 mg) or 0.05 mg / eye (equivalent to a human dose of 10 mg). Subsequently, at both dose levels, the drug was eliminated from the vitreous humor relatively quickly, consistent with a half-life of approximately 12 hours. In cynomolgus monkeys, FabA was also administered to both IVTs. In both dose groups of 1 mg / eye (equivalent to a human dose of 2 mg) and 5 mg / eye (equivalent to a human dose of 10 mg), the drug was distributed from the vitreous humor more slowly compared to rats, with a half-life of approximately 3 days.

[0211] In both species, the IVT PK of FabA was linear with respect to dose. Data from ocular toxicity studies in cynomolgus monkeys, where FabA was administered twice over a 28-day period at doses of 1.0 mg / eye (equivalent to a human dose of 2 mg), 2.5 mg / eye (equivalent to a human dose of 5 mg), or 5.0 mg / eye (equivalent to a human dose of 10 mg), showed that vitreous concentrations at sacrifice (i.e., 15 and 30 days after the second dose) were generally consistent with data from single IVT administrations.

[0212] In a 26-week chronic ocular toxicity study in cynomolgus monkeys dosed with FabA by IVT at 2.5 mg / eye per month (equivalent to a human dose of 5 mg), 5 mg / eye per month (equivalent to a human dose of 10 mg), or 5 mg / eye every other week (both 5 mg / eye doses were then reduced to 2.5 mg / eye and designated 5 / 2.5 mg / eye), the vitreous humor FabA concentration was quantifiable on day 169 in all animals that received FabA through day 184, and in all animals, it was below the quantification limit (BQL) on days 242 / 243 after a 10-week drug-free recovery period. The vitreous humor FabA concentration showed high variability, and there were no clear differences or trends between treatment groups or genders.

[0213] Pharmacokinetics of FabA in serum After a single IVT dose, serum concentrations were much lower than vitreous humor concentrations, with C max serum / C max vitreous humor being approximately 0.003 in rats and 0.000001 in cynomolgus monkeys. In cynomolgus monkeys dosed twice with FabA by bilateral IVT over a 28-day period, serum concentrations were low, with the highest mean peak concentration (C max ) of 10.1 ng / mL, which was observed after the second 5 mg / eye (equivalent to a human dose of 10 mg) IVT dose. When FabA is distributed from the vitreous humor to the serum compartment, FabA can bind to C1q or remain in a free form and be quantifiable in the assay, being non-quantifiable (i.e., <1.25 ng / mL) in the 1 mg / eye group and having mean C max values of 3.3 and 10.1 ng / mL, respectively, in the 2.5 and 5.0 mg / eye groups, resulting in low FabA serum concentrations.

[0214] In contrast, after IV administration of FabA at a dose of 10 mg / kg, in the two cynomolgus monkeys tested, the maximum FabA concentrations were 13,800 and 17,000 ng / mL, and the concentrations then decreased very rapidly, consistent with an approximately 2-hour half-life as expected for Fab fragments.

[0215] When comparing serum FabA concentrations after systemic intravenous administration of Mab2 at a dose of 200 mg / kg once a week for 4 weeks with serum FabA concentrations after two bilateral intravenous trials (5 mg / eye) over a 28-day period, serum FabA exposure was significantly lower (FabA / Mab2 Cmax ratio of 0.00000701).

[0216] In a 26-week chronic ocular toxicity study in cynomolgus monkeys, FabA was administered via in vitro at doses of 2.5 mg / eye monthly, 5 / 2.5 mg / eye monthly, or 5 / 2.5 mg / eye every other month. Systemic serum exposure to FabA was low and consistent with the local administration route. Serum FabA concentrations did not exceed 86.3 ng / mL at day 85, and did not exceed 60.8 ng / mL at day 169 after the last dose. Maximum serum FabA concentrations were observed 24–48 hours post-administration, regardless of dose level / regime and evaluation day. Serum FabA half-life (T1 / 2) values ​​were only calculable / reportable in a small number of animals in the 5 / 2.5 mg / eye every other week group, ranging from 49.9–143 hours on all evaluation days, indicating potential distribution from the ocular cavity to the serum. With monthly repeated IVT administration of 2.5 mg / eye, there was little accumulation of FabA in serum. However, in this group, the calculable serum FabA concentration increased on each subsequent evaluation day after day 1. In all other groups, accumulation from day 1 could not be determined due to changes in dosage levels after day 57. The area under the curve (AUC[0-t]) ratio to time "t" at day 169 / 85 ranged from 0.0407 to 0.664 for males and females administered 5 / 2.5 mg / eye once a month, and from 0.132 to 7.15 for males and females administered 5 / 2.5 mg / eye every other week. When comparing the sex-adjusted mean systemic exposures of FabA and Mab2 after chronic 26-week administration in cynomolgus monkeys, the FabA exposure AUC0-t (1,230 hr*ng / mL or 1.23 hr*μg / mL) after bilateral IVT administration of 5 / 2.5 mg / eye every other week was significantly lower than the AUC0-t (3,150,000 hr*μg / mL) obtained after weekly systemic IV administration of Mab2 at 200 mg / kg (0.000000073 FabA / Mab2 C max (Ratio, AUC0-t ratio of 0.00000039).

[0217] Pharmacodynamics of C1q in the eye The mean vitreous free C1q concentration in control animals was 40.3 ng / mL. In contrast, in cynomolgus monkeys that received a single IVT dose of either 1 mg / eye (equivalent to a human dose of 2 mg) or 5 mg / eye (equivalent to a human dose of 10 mg), the vitreous free C1q concentration remained below the detection limit (<1.953 ng / mL) throughout the study period (30 days), indicating complete C1q suppression. In monkeys that received FabA in a total of two doses every 28 days, C1q remained suppressed for 15 days after the second dose at all three doses (i.e., 1, 2.5, and 5 mg / eye administered twice every 28 days). 30 days after the second FabA dose, C1q remained below the detection limit in some eyes, but not all.

[0218] Fifteen days after the second 5 mg / eye IVT administration, >80% of C1q was bound to FabA in the retina, choroid, and optic disc. Thirty days after the second 5 mg / eye administration, C1q remained suppressed only in the retina and choroid.

[0219] In a 26-week chronic ocular toxicity study in cynomolgus monkeys, FabA was administered via in vitro at doses of 2.5 mg / eye monthly, 5 / 2.5 mg / eye monthly, or 5 / 2.5 mg / eye every other week. In all groups receiving FabA, a decrease in vitreous fluid C1q concentration was observed at necropsy. Animals that underwent a rest period from administration and / or a 10-week recovery period without administration showed restored vitreous fluid C1q concentrations at necropsy and post-recovery necropsy, respectively, and were comparable to the control group.

[0220] Inhibition of serum C1q and serum hemolysis Following bilateral in vitro therapy (IVT) administration of 5 mg / eye (equivalent to a human dose of 10 mg) to cynomolgus monkeys, C1q-dependent serum hemolysis was inhibited by approximately 50–80%, lasting for approximately 24–48 hours after the first IVT administration, and up to 96 hours after the second FabA administration, after which it returned to baseline.

[0221] Following a single IV dose of 10 mg / kg to cynomolgus monkeys, maximum inhibition of C1q-dependent serum hemolysis was reached within 1 hour. This maximum inhibition was maintained for approximately 24 hours, returning to baseline 120 hours after FabA administration. Serum-free C1q also decreased rapidly but did not return to baseline levels by 120 hours, suggesting that some FabA remained bound to circulating C1q throughout this timeframe.

[0222] toxicology The safety of FabA is supported by a comprehensive nonclinical ophthalmic toxicology program designed to validate the use of FabA for in vitro administration in clinical trials. Initial single-dose studies with FabA were conducted in rats and cynomolgus monkeys, and no ophthalmic toxicity was observed in either species. Based on similar findings in rats and monkeys, as well as in vitro pharmacological data, which showed monkeys to be more important than rats, and sequence homology data, cynomolgus monkeys were selected for repeated-dose ophthalmic toxicity studies of FabA.

[0223] Repeated-dose ocular toxicity studies included ophthalmoscopic examination (OE), IOP, electroretinography (ERG), ocular histopathology, and serum and intravitreous FabA measurements for TK analysis. Furthermore, the PD characteristics of FabA were determined by measuring intravitreous C1q (all in repeated-dose studies), ocular tissue (two-dose study), and inhibition of C1q-dependent hemolysis in serum (two-dose study).

[0224] Single-dose toxicity In vitro administration of FabA was well-tolerated in single-dose ocular toxicity studies (in rats and cynomolgus monkeys). In these studies, the NOAELs in rats and cynomolgus monkeys were estimated to be 0.05 mg / eye (equivalent to a human dose of 10 mg) and 5 mg / eye (equivalent to a human dose of 10 mg), respectively. These were the maximum doses evaluated in each study and were equivalent to the doses adjusted for vitreous volume (0.02 mL in rats and 2 mL in monkeys) (2.5 mg / mL).

[0225] Ocular toxicity study of single GLP-mediated intravitreal injection of FabA in Sprague Dolly rats. In this single-dose GLP rat ocular toxicity study, young adult male rats were administered either vehicle or FabA to both eyes via IVT injection at doses of 0.01 mg / eye (equivalent to a human dose of 2 mg) and 0.05 mg / eye (equivalent to a human dose of 10 mg), respectively. FabA-treated animals were sacrificed on day 1 (6 hours after administration), day 3, day 7, day 10, day 20, and day 30, while all vehicle control animals were sacrificed on day 30. All animals survived until scheduled necropsy.

[0226] This study included standard safety parameters. Blood samples were collected at slaughter, and vitreous samples were obtained at slaughter for TK analysis. In addition, ophthalmic examination (OE), including IOP, and ocular histopathology were evaluated.

[0227] No FabA-related changes were observed in any of the safety parameters evaluated, including OE, IOP, and ocular histopathology.

[0228] Vitreous exposure to FabA was confirmed by TK in animals treated with both 0.01 mg / eye (equivalent to a human dose of 2 mg) and 0.05 mg / eye (equivalent to a human dose of 10 mg) from 6 hours (initial collection) to 144 hours after administration. Serum exposure to FabA was confirmed by TK in animals treated with 0.01 and 0.05 mg / eye (only from 2 to 48 hours after administration).

[0229] No adverse reactions associated with FabA were observed at any dose level, including the maximum dose evaluated in this study, which was 0.05 mg / eye. Based on these results, the NOAEL (No Observed Adverse Effect Level) was 0.05 mg / eye (2.5 mg / mL intravitreous).

[0230] Ocular toxicity study of FabA administered via intravitreal injection (non-GLP single dose) in cynomolgus monkeys. In this single-dose non-GLP cynomolgus monkey ocular toxicity study, young adult female cynomolgus monkeys were administered either vehicle or FabA bilaterally by IVT injection at doses of 1 mg / eye (equivalent to a human dose of 2 mg) and 5 mg / eye (equivalent to a human dose of 10 mg). FabA-treated animals were sacrificed on day 1 (6 hours after administration), day 3, day 7, day 10, day 20, and day 30. All vehicle control animals were sacrificed on day 30, and all animals survived until scheduled necropsy.

[0231] This study evaluated standard safety parameters, including ocular edema (OE), intraocular ophthalmopathy (IOP), and ocular histopathology. Furthermore, blood samples were collected throughout the study, and vitreous samples were analyzed at the time of sacrifice for thyroid kinesthetic (TK) and peritoneal pedicle (PD).

[0232] Changes associated with FabA were limited to non-inflammatory, non-adverse findings. These findings included histiocyte infiltration in the uvea and mild basophilia in the 1 mg / eye dose group. Findings at the 5 mg / eye dose consisted of histiocyte infiltration in the uvea and minimal to mild basophilia.

[0233] No FabA-related changes were observed in OE and IOP. No adverse events thought to be FabA-related were observed at any dose level, including 5 mg / eye (the maximum dose evaluated) in this study. Based on these results, the NOAEL was considered to be 5 mg / eye (2.5 mg / mL intravitreous).

[0234] Vitreous exposure to FabA was confirmed by TK in all treated animals throughout the study (30 days). Serum exposure to FabA was absent at 1 mg / eye and low and transient at 5 mg / eye, not exceeding 6 ng / mL (LLOQ 1.25 ng / mL). Clq was not present in the vitreous humor of any FabA-treated animals throughout 30 days.

[0235] Repeated-dose toxicity study In the repeated-dose ocular toxicity study, FabA was administered by IVT injection at least once every 4 weeks. Repeated dosing of FabA in cynomolgus monkeys had good tolerance. In the first repeated-dose GLP ocular toxicity study, the NOAEL in cynomolgus monkeys was the maximum dose evaluated, 5 mg / eye (equivalent to a human dose of 10 mg) in twice-monthly dosing. In the 26-week chronic ocular toxicity study in cynomolgus monkeys, it was determined that the adverse eye changes were associated with the two-injection procedure and / or were ADA-mediated and not a direct effect of the IVT administration of FabA. Therefore, the NOAEL was determined to be 2.5 mg / eye (equivalent to a human dose of 5 mg) in dosing of 13 times or 7 times at every other week or once a month, respectively, in cynomolgus monkeys.

[0236] 6-week GLP repeated-dose ocular toxicity study of FabA by intravitreal injection in cynomolgus monkeys This study included standard safety parameters and blood samples were collected throughout the study. Vitreous samples at sacrifice for TK and PD analysis and optic nerve sections at sacrifice for TK and PD analysis were also collected. Additionally, OE, IOP, ERG and ocular histopathology were also evaluated.

[0237] The findings of FabA determined to be non-harmful were limited to one female at a single high dose (2.5 mg / eye) (equivalent to a human dose of 5 mg), and this female had minimal basophilic / blue staining of the vitreous without inflammation (referred to as an increase in basophils). Importantly, no changes related to FabA were observed in OE, IOP and ERG. Exposure to FabA in the vitreous of all treated animals was confirmed by TK during the study and recovery (30 days after the last dose).

[0238] Serum exposure was not measurable at 1 mg / eye (equivalent to a human dose of 2 mg), was low and transient (12 - 48 hours after the first dose, 6 - 168 hours after the last dose) at 2.5 mg / eye (equivalent to a human dose of 5 mg), and did not exceed 8 ng / mL (LLOQ 1.25 ng / mL). When the FabA concentration was approximately 100 ng / mL, it was confirmed by PD that there was no C1q in the vitreous of all treated animals. ADA to FabA was detected in animals dosed at 1 mg / eye (equivalent to a human dose of 2 mg) (6 out of 12 animals) and 2.5 mg / eye (equivalent to a human dose of 5 mg) (7 out of 12 animals), but there was no clear effect of ADA on FabA exposure in serum or vitreous.

[0239] 6 - week GLP repeated dose ocular toxicity study of FabA by intravitreal injection in cynomolgus monkeys In this 6 - week GLP cynomolgus monkey ocular toxicity study, young adult male and female cynomolgus monkeys were administered vehicle or FabA by bilateral intravitreal injection at a dose of 5 mg / eye (equivalent to a human dose of 10 mg) every 4 weeks (days 1 and 29), followed by a 4 - week recovery period. All major study animals were sacrificed on day 44 and all recovery animals were sacrificed on days 59 / 60. All major study animals and recovery animals survived until scheduled necropsy.

[0240] This study included standard safety parameters (excluding systemic histopathology), blood samples were collected throughout the study, and vitreous samples at sacrifice were also collected for TK and PD analysis. Samples of ADA and aqueous humor were collected and stored. Additionally, OE, IOP, ERG, and ocular histopathology were also evaluated.

[0241] No changes related to FabA were observed in any safety parameter evaluated, including OE, IOP, ERG, and ocular histopathology. Minimal to mild basophilic / blue staining of the vitreous (termed basophil increase), without inflammation, was observed in both treated and control animals and was thus considered not related to FabA.

[0242] Exposure to FabA in the vitreous humor of all treated animals was confirmed by TK during the study and recovery period (30 days after the last dose). The absence of FabA in the serum and vitreous humor of control animals was confirmed. In all treated primary test animals, the absence of C1q in the vitreous humor was confirmed by PD on day 44. On day 59, two out of four recovered animals had measurable C1q in the vitreous humor. ADA of FabA was detected in animals (9 out of 10 animals) in the 5 mg / eye dose group (equivalent to the human dose of 10 mg), but there was no clear effect of ADA on serum or vitreous FabA exposure.

[0243] >80% inhibition of C1q-dependent hemolysis was achieved 24–48 hours after FabA administration, after which it returned to baseline.

[0244] On day 44, C1q density in the retina, choroid, and optic nerve head decreased significantly, and on day 59, it continued to decrease in the retina and choroid, but not in the optic nerve head.

[0245] In this study, no adverse events thought to be related to FabA were observed at any dose level, including 5 mg / eye (equivalent to a human dose of 10 mg) (the maximum dose evaluated). Based on these results, the NOAEL was 5 mg / eye (equivalent to a human dose of 10 mg) (2.5 mg / mL in the vitreous humor).

[0246] A 26-week GLP repeated-dose ocular toxicity study of FabA administered intravitreally in cynomolgus monkeys, with a 10-week recovery period. In this 26-week GLP cynomolgus monkey ocular toxicity study, young adult male and female cynomolgus monkeys were administered bilaterally by IVT injection of Vehicle or FabA at doses of 2.5 mg / eye (equivalent to a human dose of 5 mg) once monthly, 5 mg / eye (equivalent to a human dose of 10 mg) once monthly, and 5 mg / eye every other week, followed by a 10-week recovery period. A single 50 μl injection equivalent to 2.5 mg / eye, or two injections totaling 100 μL (two 50 μL injections 10 minutes apart, equivalent to 5 mg / eye), were administered every two weeks (13 administration periods) or every four weeks (7 administration periods). All primary test animals were sacrificed on day 184, and all recovered animals were sacrificed on days 242 / 243. All primary test animals and recovered animals survived until scheduled necropsy.

[0247] No-administration days or discontinuations occurred in the control group, the group receiving 5 mg / eye once a month (equivalent to a human dose of 10 mg), and the group receiving 5 mg / eye every other week. The two injections in these groups were discontinued due to adverse findings detected by OE, which were considered to be related to the procedure and the high dose volume. From day 71 of the study, the group receiving 5 mg / eye once a month received 2.5 mg / eye once a month (equivalent to a human dose of 5 mg) (referred to as 5 / 2.5 mg / eye once a month), and the group receiving 5 mg / eye every other week received 2.5 mg / eye every other week (referred to as 5 / 2.5 mg / eye every other week). In these administration groups (including the control group), no-administration days continued after discontinuation of the two injections. The discontinuation of the two injections was related to the procedure and / or ADA, as described below. There were no no-administration days in the group receiving 2.5 mg / eye once a month (low-dose group).

[0248] This study included standard safety parameters (excluding systemic histopathology), and blood samples were collected throughout the study, as were vitreous samples at the time of sacrifice for TK and PD analysis. ADA and aqueous humor samples were collected and stored. Furthermore, OE, IOP, ERG, ocular histopathology, and immunohistochemistry (IHC) for the detection of immune complexes deposited within the eyeball were evaluated.

[0249] No FabA-related changes were observed in body weight, food intake, electroretinography, intraocular pressure measurement, or clinical condition.

[0250] Clinical signs and examination findings of the eye thought to be associated with FabA were limited to opacity of the eyeball (presumably due to opacity in the anterior chamber, lens capsule, and / or posterior chamber) and the presence of cells and / or pigments. The presence of these findings in animals in which ADA was not detected in the serum (4 out of 12 animals in Group 2, 2 out of 12 animals in Group 3, and 2 out of 12 animals in Group 4) indicates an association with FabA. Findings considered to be associated with ADA and potentially immune complex deposition tended to be more severe and included aqueous humor flare, the presence of vitreous opacity, altered pupillary light reflex, and attenuation of retinal vascularity.

[0251] Following in vitro administration of FabA to male and female monkeys, systemic exposure, as measured by serum FabA concentration, was transient and low, not exceeding 86.3 ng / mL after administration on day 85 or 60.81 ng / mL after the final administration on day 169. In almost all treated animals that received FabA, exposure to FabA was confirmed by TK throughout day 169, which was consistent with the undetectable vitreous C1q, except in some animals that had a no-administration period. On days 242 / 243 after a 10-week recovery period without administration, vitreous FabA concentrations were unmeasurable, while C1q concentrations were detectable in all treated groups. The absence of FabA in the serum and vitreous humor of control animals was confirmed.

[0252] The presence of anti-FabA antibodies in serum samples was confirmed in 4 out of 12 animals in Group 1 (control), 8 out of 12 animals in Group 2, 10 out of 12 animals in Group 3, and 10 out of 12 animals in Group 4. Positivity in 2 animals in Group 1 was confirmed once per animal after day 1, while ADA positivity in FabA-treated animals was identified at 3 or 4 or more time points (a total of 4 or 5 samples were collected for the primary test animals and recovered animals, respectively). There was no clear effect of ADA on serum or intravitreous FabA exposure.

[0253] At the final euthanasia on day 184, microscopic changes consistent with an ADA-mediated immune response to FabA were observed in the right eye of the monthly and bi-weekly 5 / 2.5 mg / eye administration groups (moderate and high dose groups, respectively). Intraocular changes associated with inflammation included mild mixed cell infiltration in the ciliary body and vitreous cavity, minimal to moderate fibrosis in the vitreous cavity (severity proportional to administration frequency), and minimal to mild posterior lens degeneration (severity proportional to administration frequency). Minimal perivascular mononuclear cell infiltration in the posterior retina was also observed in one female animal in each of the bi-weekly and monthly 5 / 2.5 mg / eye treatment groups. Minimal to moderate mononuclear cell infiltration was also observed in the limbus around the eyeball of animals administered 5 / 2.5 mg / eye bi-weekly and monthly, with severity proportional to administration frequency.

[0254] At euthanasia following recovery on day 242 / 243, microscopic changes in the right eye associated with the ADA-mediated immune response to FabA were limited and mild in the group receiving monthly 5 / 2.5 mg / eye, while further changes persisted or occurred in the group receiving bi-weekly 5 / 2.5 mg / eye. Minimal histiocyte infiltration in the vitreous cavity and uvea, as well as increased basophilia in the vitreous cavity, were limited during the recovery phase in animals administered bi-weekly and / or monthly FabA 5 / 2.5 mg / eye. These changes were similar to those observed in control animals at final autopsy and were considered to be related to mild inflammation / destruction of the anterior vitreous humor associated with the IVT injection procedure. However, the persistence of these changes after the recovery period, their resolution in control animals during the recovery period, and the presence of more severe inflammatory changes at final necropsy in animals administered 5 / 2.5 mg / eye of FabA every other week and monthly suggested that these changes during the recovery period may indicate the resolution of inflammation associated with the ADA-mediated immune response to FabA, rather than residual effects from the injection procedure. Minimal mononuclear cell infiltration in the limbus periocularly persisted in both groups administered 5 / 2.5 mg / eye every other week and monthly.

[0255] In the group receiving 5 / 2.5 mg / eye every other week, minimal mixed cell infiltration and fibrosis persisted in the vitreous cavity, while mild cellular decline and hemosiderin pigmentation occurred in the retina. Minimal perivascular mononuclear cell infiltration in the optic disc was also observed in the retina in the 5 / 2.5 mg / eye group. These changes were considered secondary to the ADA-mediated response to FabA.

[0256] Pathologically harmful microscopic changes were considered secondary to ADA-mediated inflammation and included intravitreal fibrosis, lens degeneration, and retinal cellular degeneration in the bi-weekly and monthly 5 / 2.5 mg / eye dose groups.

[0257] In the group receiving 2.5 mg / eye per month, no microscopic changes related to FabA were observed in either final or post-recovery euthanasia.

[0258] Immunohistochemistry was performed on 2 out of 12 animals from Group 1, 4 out of 12 animals from Group 3, and 6 out of 12 animals from Group 4. The presence of granular deposits containing FabA, simian IgG, IgM, and / or C3, immunohistochemically detected in the left eyes of 4 out of 10 treated animals in the moderately dosed group selected for IHC, i.e., the group receiving 5 / 2.5 mg / eye monthly (2 out of 4 animals), and the high dosed group receiving 5 / 2.5 mg / eye every other week (2 out of 6 animals), was revealed by evaluation. These intravascular wall deposits were associated with perivascular inflammatory cell infiltration and were similar to the deposits observed by hematoxylin-eosin evaluation of the right eyes. Other microscopic changes observed in the right eyes were consistent with secondary changes associated with the immune response to this FabA in monkeys. Although ocular immune complex deposits were not observed in all animals selected for immunohistochemistry, including some animals with serum ADA negativity, this was not unexpected because the identification of deposits can vary with tissue sectioning, and serum ADA does not always correlate with the immune complex pathology in animals with microscopic evidence. Additionally, in some animals with multiple drug-free days, there is a possibility that ADA and / or immune complexes were cleared prior to analysis. The presence of deposits confirmed immunohistochemically represents the most compelling evidence that similar pathogenetically consistent pathologies observed in the right eyes, even in a subset of animals, appear to be associated with the immune response to FabA.

[0259] In this 26-week chronic ocular toxicity study in cynomolgus monkeys, it was determined that the adverse ocular changes were associated with the two injection procedures and / or were mediated by ADA and not a direct effect of the IVT administration of FabA. Therefore, the NOAEL was determined to be 2.5 mg / eye (equivalent to a human dose of 5 mg) with administration every other week or monthly for 13 or 7 times, respectively, in cynomolgus monkeys.

[0260] Example 2: Evaluation of FabA in Clinical Trials FabA is a sterile isotonic solution for in vitro vasoconstriction (IVT) injection. A Phase 1, first-in-human, open-label, dose-escalation study (FabA-GLA-01) was conducted to evaluate the initial safety and tolerability of a single IVT injection of FabA in patients with primary open-angle glaucoma.

[0261] A Phase 1b, randomized, double-blind trial (FabA-GLA-02) was conducted to evaluate the safety and tolerability of repeated IVT injections of FabA in patients with primary open-angle glaucoma.

[0262] Results from both trials showed that single-dose (1-5 mg / eye) and repeated-dose (2.5 and 5 mg / eye, administered 4 weeks apart) of FabA via IVT were well-tolerated in glaucoma patients, and no serious adverse events (AEs) were reported. In these trials, ocular AEs in patients treated with FabA included conjunctival hyperemia, conjunctival hemorrhage, and ocular irritation, and occurred only in the treated eye. In the phase 1b trial, ocular AEs in patients in the sham group included eye pain, foreign body sensation, conjunctival hyperemia, and blurred vision. No systemic AEs thought to be related to FabA IVT treatment occurred.

[0263] A single in vitro injection of FabA 2.5 mg (equivalent to a human dose of 5 mg) and 5 mg (equivalent to a human dose of 10 mg) inhibited free C1q in aqueous humor for at least 29 days (Study FabA-GLA-02).

[0264] Pharmacokinetics and pharmacodynamics in humans Pharmacokinetics and pharmacodynamics of the eye FabA-GLA-02 was a phase 1b study that evaluated PK and PD by collecting aqueous humor. Participants received two in vitro injections of sham, 2.5 mg / eye of FabA (equivalent to a human dose of 5 mg), or 5 mg / eye (equivalent to a human dose of 10 mg) of FabA, 29 days apart. In this study, aqueous humor was collected before administration, 29 days after the first FabA administration, and before the second administration. Free FabA was detected in the aqueous humor of all treated patients on day 29 (D29). Simultaneously, both 2.5 mg / eye and 5 mg / eye dose levels of FabA inhibited free C1q in the aqueous humor for at least 29 days (Figure 6).

[0265] Systemic pharmacokinetics and pharmacodynamics FabA-GLA-01 was a single-dose phase 1 study in which serum FabA and C1q were collected before administration and 3 hours after administration. FabA-GLA-02 was a multi-dose phase 1b study in which serum, FabA, and C1q were collected before and 3 hours after each of two administrations at 29-day intervals. At any dose level tested in the phase 1 or phase 1b clinical trials, FabA was not typically detectable in the systemic circulation after a single or repeated IVT injection. Similarly, no changes in circulating free C1q were detected in either study.

[0266] As described below, the 5 mg / eye dose level (equivalent to a human dose of 10 mg) was well-tolerated in the FabA clinical trial when administered as a single dose or in two doses at 29-day intervals. As mentioned above, in the Phase 1b trial, single doses of FabA at 2.5 mg (equivalent to a human dose of 5 mg) and 5 mg (equivalent to a human dose of 10 mg) inhibited free C1q in aqueous humor for at least 29 days (Figure 6).

[0267] Safety and effectiveness Phase 1 dose escalation study (FabA-GLA-01) This was a Phase 1, open-label, dose-escalation study evaluating the safety / tolerability and pharmacokinetics of a single IVT injection of FabA in patients with primary open-angle glaucoma. Eligible patients were adults who had a mean deviation of 3–18 dB on reliable visual field testing using the Humphrey Field Analyzer-Swedish Interactive Threshold Algorithm (HFA-SITA) 24-2 fast algorithm, with a cutoff of 33% for poor fixation and a cutoff of 33% for false positive response rate, and who had IOP <21 mmHg in the test eye during a stable IOP treatment regimen for ≥4 weeks prior to administration. Nine patients were assigned to three cohorts, with three patients enrolled per cohort as follows: Cohort 1 = 1.0 mg / eye, single dose (0.02 mL) x 1 dose Cohort 2 = 2.5 mg / eye, single dose (0.05 mL) x 1 dose Cohort 3 = 5.0 mg / eye, single dose (0.10 mL) x 1 dose Following screening, three eligible patients were enrolled in the lowest open cohort and then in subsequent cohorts, which would only be initiated after tolerability and short-term safety were demonstrated at the aforementioned low doses. All patients in each cohort were required to complete a minimum 15-day safety observation period before being able to receive injections in the next cohort. No dose-limiting toxicities (DLTs) were reported during the study.

[0268] Nine patients were enrolled, received treatment, and completed the trial.

[0269] safety TEAEs (Triaded Eye Events) in the eye included conjunctival hyperemia (all dose levels), conjunctival hemorrhage (2.5 mg / eye only), and eye irritation (1 mg / eye only), which occurred only in the test eye.

[0270] The only systemic TEAE experienced in this study was sinusitis. • All TEAEs were mild in severity. No serious or significant TEAEs were observed. No patients discontinued treatment or withdrew from the trial due to TEAEs. • In all nine patients, IOP returned to normal (within 5 mmHg of the IOP immediately before injection, or <21 mmHg) within 30 minutes. • No patients demonstrated any evidence of anti-FabA antibodies.

[0271] Overall summary / conclusion: In this study, a single in vitro dose of FabA was well-tolerated up to 5 mg / eye in patients with stable glaucoma. Reported ocular adverse events (AEs) were similar to those reported with in vitro doses of approved drugs. No safety signals were observed for FabA.

[0272] FabA is not normally detectable in systemic circulation, and no changes in circulating free C1q were detected after a single IVT administration.

[0273] Phase 1b (FabA-GLA-02) This was a double-blind, randomized, sham-controlled trial to evaluate two dose levels of FabA administered as repeated IVT injections versus sham injection in patients with primary open-angle glaucoma. Eligible patients were adults who had a mean deviation of -3 to -24 dB in the test eye, had IOP <21 mmHg at screening and day 1, were on a stable IOP treatment regimen of ≥4 weeks prior to injection, and had no expected changes to their IOP treatment regimen during the trial. Patients received two injections 4 weeks apart and were followed for a total of 12 weeks for safety, tolerability, PK, PD, immunogenicity assessment and ongoing exploratory evaluation. Patients were randomly assigned to one of three cohorts (a plan with 5 patients per cohort) as follows (1:1:1). • Dosage level 1 = 2.5 mg / eye, single dose (0.05 mL) x 2 doses • Dosage level 2 = 5.0 mg / eye, single dose (0.10 mL) x 2 doses • Siam = 0 mg / eye x 2 doses Eighteen patients were randomized (7 to the 2.5 mg FabA group, 5 to the 5.0 mg FabA group, and 6 to the sham group), and 17 patients received treatment. One patient in the 2.5 mg group was randomized but did not receive treatment. Sixteen patients completed the trial.

[0274] safety Ocular TEAEs experienced by patients treated with FabA included conjunctival hyperemia (2.5 and 5 mg / eye), conjunctival hemorrhage (5 mg / eye only), and ocular irritation (5 mg / eye only). None of these TEAEs were experienced by patients in the sham group. Ocular TEAEs in patients in the sham group included eye pain, foreign body sensation, conjunctival hyperemia, and blurred vision, each occurring in one patient.

[0275] Systemic TEAEs were observed during the trial, but none were attributed to the study treatment by the researchers. • All TEAEs were mild in severity. • All but one of the reported TEAEs occurred after the first dose and before the second dose of the study treatment. No serious or significant TEAEs were observed. No patients discontinued treatment or withdrew from the trial due to TEAEs. • In 16 of the 17 patients, IOP returned to normal (<21 mmHg) within 30 minutes of IVT injection, and in the remaining patients, within 45 minutes. Of the 11 patients who received intravitreal FabA, 6 tested were positive at least at one time point. One patient was ADA-positive with a gradual increase in titer over time, while the remaining 5 patients were positive at all time points, including before administration, and their titers did not change over time. One sham patient was ADA-positive at all time points, including before administration, and their titers did not change over time. In summary, these data suggest that the relationship between ADA measurements and FabA administration is unclear.

[0276] Overall summary / conclusion: In patients with stable glaucoma, two in vitro vasoconstrictions (IVT) of FabA at 4-week intervals were well tolerated up to a maximum dose of 5 mg / eye. Reported ocular adverse events (AEs) were similar to those reported with IVT of approved drugs. No safety signals were observed for FabA in this study.

[0277] A single in vitro dose of FabA (2.5 and 5 mg / eye) inhibited free C1q in aqueous humor for at least 29 days.

[0278] Example 3: Phase 2, multicenter, randomized, parallel-group, double-blind, 4-arm, sham-controlled study to evaluate the efficacy, safety, and tolerability of FabA administered by intravitreal injection in patients with geographic atrophy (GA) secondary to age-related macular degeneration (AMD). basis overview This trial will be conducted in patients with GA secondary to AMD. The objective of the trial is to determine whether intravitreal (IVT) injections of FabA once monthly (EM) or once every two months (EOM) for 12 months reduce the proliferation rate of GA lesions. The trial will consist of a 30-day screening period, a 12-month treatment period, and a subsequent 6-month (no treatment) follow-up period. The total patient participation period will be 19 months. Patients will visit the clinic monthly during the 12-month treatment period for treatment and / or safety evaluation.

[0279] Approximately 240 patients were enrolled, and about 204 patients were randomly assigned to one of four treatment arms so that the primary analysis (based on modified intention to treat [ITT]) could be evaluated at month 12.

[0280] Intervention group and duration: The trial intervention was randomized (2:2:1:1). Patients were assigned to one of the following treatment arms. Dosage levels were fixed and not changed. Arm 1 = FabA 5.0 mg / eye (0.10 mL) once a month (EM) for 12 months (12 doses) Arm 2 = FabA 5.0 mg / eye (0.10 mL) once every other month (EOM), for 12 months (6 doses). • Arm 3 = Sham injection with EM for 12 months (12 Sham injections) • Arm 4 = Sham injections at EOM for 12 months (6 Sham injections)

[0281] injection FabA / Sham administration is completed by the physician administering the injection using sterile techniques.

[0282] All patients randomized to FabA will receive 5.0 mg / eye in a fixed volume of 0.10 mL via IVT, once a month or every other month, for 12 months.

[0283] After injection Immediately after administering the medication, the physician administering the injection should assess whether the patient can see hand valve vision or central retinal artery perfusion. If necessary, rule out other causes of visual impairment, such as vitreous hemorrhage. If necessary, perform digital massage until hand valve vision or central retinal artery perfusion is observed, and administer additional topical / oral IOP-reducing medication.

[0284] IOP (intraocular pressure) is evaluated only in the test eye 30 minutes after drug administration. If it is elevated, it is evaluated every 15 minutes thereafter until IOP < 25 mmHg.

[0285] Pharmacokinetics, pharmacodynamics, and immunogenicity Blood samples for PK evaluation (FabA serum concentration) and PD evaluation (serum C1q concentration and plasma concentrations of other biomarkers) will be collected during the patient visit, 30 minutes before administration and within 3 hours (±15 minutes) after administration.

[0286] During the patient's clinic visit, collect a sample for immunogenicity testing (ADA) before the injection. Additionally, collect another ADA sample in the second week, either at the clinic or during a home visit.

[0287] Pharmacokinetics: This study requires serum. A blood sample will be collected to measure the serum concentration of FabA.

[0288] Pharmacodynamics: This test requires serum and plasma. Serum C1q concentration and plasma concentration of exploratory complement biomarkers are analyzed.

[0289] Immunogenicity: This test requires serum. Immunogenicity is assessed by analysis of anti-drug (FabA) antibodies (ADA) in the serum.

[0290] Procedure for intravitreal injection Preparation of FabA From the sterile vial of FabA, withdraw the total volume of FabA (approximately 0.3 mL) using a sterile 1.0 cc syringe equipped with a 19 gauge x 1 1 / 2 inch, 5 micrometer filter needle.

[0291] Replace the filter needle with a 30 gauge x 1 / 2 inch injection needle. Immediately before injection, drain any excess volume of FabA from the syringe, leaving only the required injection volume.

[0292] The dosage volume of FabA should be fixed at 0.10 mL, administered once a month (EM) for 12 months (12 doses), or once every two months (EOM) for 12 months (6 doses).

[0293] Preparation for intravitreal injection 1. Examine the test eye. 2. Before injection, measure and record the preoperative intraocular pressure (IOP) of the test eye. IOP should only be measured in the test eye. To proceed, the IOP must be ≤21 mmHg. If it is >21 mmHg, the researcher may, at their discretion, change the FabA injection schedule and manage the IOP. 3. If necessary, apply one drop of 2.5% phenylephrine chloride eye drops to the test eye 30 minutes before the injection to enable visualization of the posterior pole after injection. 4. Immediately before injection: • Have the patient lie down on the examination chair with their neck adequately supported.

[0294] Intravitreal injection 1. Handwashing, sterile gloves, and a surgical mask are required for injections. 2. Apply 0.5% topical propalacine to the test eye. 3. Apply 10% povidone-iodine to the eyelashes and eyelid margins. Avoid extensive eyelid massage before or after injection to avoid exposing the meibomian glands. 4. During the procedure, pull the eyelid away from the target injection site. Use of a microscope is recommended. 5. Apply 5% povidone-iodine to the conjunctival surface, including the target injection site. 6. FabA injection: Insert the needle perpendicular to the sclera between the vertical and horizontal rectus muscles, 3.5-4 mm posterior to the limbus. Immediately after withdrawing the needle, apply a sterile cotton swab applicator to the injection site to reduce vitreous reflux. 7. Sham Injection: Preparation and post-injection care for the sham injection are the same as for injections using FabA. The sham injection is performed by applying pressure to the eye using the blunt needle of an empty syringe without a needle at the typical intravitreal injection site.

[0295] After intravitreal injection 1. After the injection, the patient is required to remain at the clinic for eye evaluation and safety follow-up. 2. Immediately assess hand valve vision or central retinal artery perfusion and rule out other causes of visual impairment, such as vitreous hemorrhage. If no other cause is found, perform digital massage until hand valve vision or central retinal artery perfusion is observed, and administer topical / oral IOP-reducing agents. 3. In the test eye, obtain an IOP measurement only 30 minutes after injection. If it is elevated, measure every 15 minutes until IOP < 25 mmHg. If the intraocular pressure exceeds 30 mmHg after 15 minutes, treatment should be administered at the physician's discretion. 4. Topical antibiotics are not necessary.

[0296] Embedding by reference Each of the patents, published patent applications, and non-patent literature cited herein is incorporated herein by reference in its entirety.

[0297] Equal parts Those skilled in the art will recognize, or can confirm by mere ordinary experimentation, many equivalents to the particular embodiments of the invention described herein. Such equivalents are intended to be covered by the following claims.

Claims

1. A composition comprising an anti-C1q antibody or antigen-binding fragment thereof for treating an ocular disease in a human patient, said composition being for intravitreal injection at a dose of about 1 mg to about 10 mg; The composition, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising HVR-L1 having the amino acid sequence of SEQ ID NO:5, HVR-L2 having the amino acid sequence of SEQ ID NO:6, and HVR-L3 having the amino acid sequence of SEQ ID NO:7, and a heavy chain variable domain comprising HVR-H1 having the amino acid sequence of SEQ ID NO:9, HVR-H2 having the amino acid sequence of SEQ ID NO:10, and HVR-H3 having the amino acid sequence of SEQ ID NO:

11.

2. The composition of claim 1, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs: 4 and 35-38, and the light chain variable domain comprises HVR-L1 having the amino acid sequence of SEQ ID NO: 5, HVR-L2 having the amino acid sequence of SEQ ID NO: 6, and HVR-L3 having the amino acid sequence of SEQ ID NO:

7.

3. The composition of claim 2, wherein the light chain variable domain comprises an amino acid sequence selected from SEQ ID NOs: 4 and 35-38.

4. The composition of any one of claims 1 to 3, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence having at least about 95% homology to an amino acid sequence selected from SEQ ID NOs: 8 and 31 to 34, and the heavy chain variable domain comprises HVR-H1 having the amino acid sequence of SEQ ID NO: 9, HVR-H2 having the amino acid sequence of SEQ ID NO: 10, and HVR-H3 having the amino acid sequence of SEQ ID NO:

11.

5. The composition of claim 4, wherein the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NOs: 8 and 31-34.

6. The composition of any one of claims 1 to 5, wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody, a humanized antibody, a human antibody, a chimeric antibody, an antibody fragment, or an antibody derivative thereof.

7. The composition of claim 6, wherein the antibody fragment is a Fab fragment, a Fab' fragment, an F(ab')2 fragment, an Fv fragment, a diabody, or a single-chain antibody molecule.

8. The composition of claim 7 , wherein the Fab fragment comprises a heavy chain Fab fragment of SEQ ID NO:39 and a light chain Fab fragment of SEQ ID NO:

40.

9. The composition of any one of claims 1 to 8, which is for administration once weekly, once every two weeks, once every three weeks, once a month, once every four weeks, once every six weeks, once every eight weeks, once every two months, once every ten weeks, once every twelve weeks, once every three months, or once every four months.

10. The composition of claim 9, which is for administration for at least 3 months, at least 4 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 12 months.

11. The composition described in claim 9, which is intended to be administered for 12 months.

12. 12. The composition of any one of claims 1-11, wherein the dose comprises about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, or about 10 mg of the anti-C1q antibody.

13. 13. The composition of any one of claims 1-12, wherein the dose comprises about 1 mg to about 2.5 mg, about 2.5 mg to about 5 mg, about 5 mg to about 7.5 mg, or about 7.5 mg to about 10 mg of the anti-C1q antibody.

14. The composition according to any one of claims 1 to 13, wherein the eye disease is glaucoma or age-related macular degeneration.

15. The composition of claim 14, wherein the age-related macular degeneration is geographic atrophy.

16. Use of an anti-C1q antibody or antigen-binding fragment thereof in the manufacture of a medicament for the treatment of an ocular disease in a human patient, the medicament being for intravitreal injection of the anti-C1q antibody or antigen-binding fragment thereof at a dose of about 1 mg to about 10 mg; The above-mentioned use, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising HVR-L1 having the amino acid sequence of SEQ ID NO:5, HVR-L2 having the amino acid sequence of SEQ ID NO:6, and HVR-L3 having the amino acid sequence of SEQ ID NO:7, and a heavy chain variable domain comprising HVR-H1 having the amino acid sequence of SEQ ID NO:9, HVR-H2 having the amino acid sequence of SEQ ID NO:10, and HVR-H3 having the amino acid sequence of SEQ ID NO:11.

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