US20160159890A1 - Methods of treatment for alzheimer's disease and huntington's disease - Google Patents

Methods of treatment for alzheimer's disease and huntington's disease Download PDF

Info

Publication number
US20160159890A1
US20160159890A1 US14/988,387 US201614988387A US2016159890A1 US 20160159890 A1 US20160159890 A1 US 20160159890A1 US 201614988387 A US201614988387 A US 201614988387A US 2016159890 A1 US2016159890 A1 US 2016159890A1
Authority
US
United States
Prior art keywords
antibody
seq
amino acid
acid residues
antibodies
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US14/988,387
Other languages
English (en)
Inventor
Arnon Rosenthal
Michael Leviten
Beth A. Stevens
Soyon Hong
Daniel Wilton
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Boston Childrens Hospital
Annexon Inc
Original Assignee
Boston Childrens Hospital
Annexon Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Boston Childrens Hospital, Annexon Inc filed Critical Boston Childrens Hospital
Priority to US14/988,387 priority Critical patent/US20160159890A1/en
Assigned to CHILDREN'S MEDICAL CENTER CORPORATION reassignment CHILDREN'S MEDICAL CENTER CORPORATION ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: HONG, Soyon, STEVENS, BETH A., WILTON, Daniel
Assigned to ANNEXON, INC. reassignment ANNEXON, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: LEVITEN, MICHAEL, ROSENTHAL, ARNON
Publication of US20160159890A1 publication Critical patent/US20160159890A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A61P21/04—Drugs for disorders of the muscular or neuromuscular system for myasthenia gravis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/06—Antiglaucoma agents or miotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/06—Antianaemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56966—Animal cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A61K2039/507—Comprising a combination of two or more separate antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/54—F(ab')2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/55—Fab or Fab'
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/734—Complement-dependent cytotoxicity [CDC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
    • G01N2333/4701—Details
    • G01N2333/4716—Complement proteins, e.g. anaphylatoxin, C3a, C5a

Definitions

  • This invention relates generally to methods of treatment for neurodegenerative diseases such as Alzheimer's disease, Alzheimer's-related diseases, and Huntington's disease, and more specifically to methods involving the inhibition of the classical pathway of complement activation.
  • a neurodegenerative disease is a disease involving cognitive disorders, such as Alzheimer's disease (AD) and Huntington's disease (HD), to name but a few. These cognitive disorders are caused by an increase in cell death processes that results in a great reduction of neuron number, behavioral changes and a general, gradual degeneration that leads to the patient's death.
  • AD Alzheimer's disease
  • HD Huntington's disease
  • Alzheimer's disease is characterized by a progressive loss of cognitive function associated with an excessive number of senile plaques in the cerebral cortex and subcortical gray matter. Alzheimer's disease accounts for more than 65% of dementias in the elderly. Pathologically, Alzheimer's disease is characterized by two brain lesions, called senile plaques and neurofibrillary tangles (NFT), which accumulate, respectively, ABeta (A ⁇ ) peptides and the microtubule-associated protein tau (MAPT). These neuropathological hallmarks have given rise to two corresponding hypothesis of the neurodegenerative process: the ⁇ /amyloid cascade hypothesis (Hardy and Selkoe 2002; Karran et al.
  • a ⁇ /amyloid cascade hypothesis the A ⁇ peptide evolves from the cleavage of the amyloid precursor protein (APP) by ⁇ / ⁇ -secretase complexes, which associate presenilins-1 or -2 (PS-1 or PS-2), leading typically to the formation of a 38, 40, or 42 amino acid peptide (A ⁇ 38, 40 or 42).
  • PS-1 or PS-2 presenilins-1 or -2
  • a ⁇ 42 is the predominant form that aggregates outside the cells forming the characteristic amyloid plaques (Yan and Wang 2008).
  • a ⁇ peptide i.e., the more C-terminal within the ⁇ -CTF transmembrane domain where g-secretase cleaves), the greater its hydrophobicity and propensity to oligomerize (Haass and Selkoe 2007).
  • Dyshomeostasis of A ⁇ , in particular, A ⁇ 42 is currently thought to be one of the earliest biomarkers of Alzheimer's disease in humans, even before clinical symptoms emerge (Sperling et al., 2011); the lowering of the A ⁇ 42 peptide precedes the rise of levels of tau and phosphorylated tau in the CSF of humans (Craig-Schapiro et al., 2009; Golde et al., 2011).
  • a ⁇ dyshomeostasis As the principal driving force in Alzheimer's disease (Tanzi 2012). Current evidence also suggests that soluble low-n A ⁇ oligomers, but not A ⁇ monomers, mediate synaptotoxicity (Walsh and Selkoe 2007). Several studies suggest that small oligomeric morphologies of A ⁇ are the primary toxic species in Alzheimer's disease (Glabe et al. 2006; Walsh et al. 2002).
  • Huntington's disease is an inherited neurodegenerative disease, characterized by motor, cognitive, behavioral, and psychological dysfunction. Similar to Alzheimer's disease, where protein aggregates are associated with disease, Huntington's disease belongs to a family of neurodegenerative diseases caused by mutations in which an expanded CAG repeat tract results in long stretches of polyglutamine (polyQ) in the encoded protein (i.e., mutant Huntington) which in Huntington's disease forms aggregates within the cell (Martindale et al., 1998).
  • polyQ polyglutamine
  • Synaptic dysfunction and loss are early manifestations of disease in human Huntington's disease with significant deficits in synaptic protein levels (a-tubulin, neurofilament, MAP-2) and subcellular distribution (neurofilament, PACSIN 1) beginning at the pre-symptomatic stage of disease (DiProspero et al., 2004).
  • This synaptic dysfunction has been explored in mouse transgenic models of Huntington's disease, confirming that decreased synapse function is an early manifestation of disease.
  • LTP long-term potentiation
  • N-methyl-D-asparate receptor antagonists e.g., Ebixa, Namenda
  • acetylcholinesterase inhibitors e.g., Razadyne, Exelon, Aricept and Cognex
  • Certain aspects of the present disclosure provide anti-C1q antibodies and methods of using such antibodies for treating or preventing Alzheimer's disease, Alzheimer's-related diseases, and Huntington's disease.
  • Certain aspects of the present disclosure are directed to methods and kits for treating or preventing Alzheimer's disease and/or Huntington's disease that include inhibiting the classical pathway of complement activation by neutralizing complement factor C1q, e.g., through the administration of antibodies, such as monoclonal, chimeric, humanized antibodies, antibody fragments, etc., which bind to C1q.
  • the present disclosure provides a method of treating or preventing Alzheimer's disease in an individual in need of such treatment, the method comprising the step of administering a therapeutically effective dose of an anti-C1q antibody.
  • the present disclosure provides an anti-C1q antibody for use in treating or preventing Alzheimer's disease in an individual in need of such treatment.
  • the present disclosure provides use of an anti-C1q antibody in the manufacture of a medicament for treating or preventing Alzheimer's disease.
  • the present disclosure provides a kit comprising an anti-C1q antibody and a package insert comprising instructions for using the antibody to treat or prevent Alzheimer's disease in an individual in need of such treatment.
  • the present disclosure provides a method of treating or preventing Huntington's disease in an individual in need of such treatment, the method comprising the step of administering a therapeutically effective dose of an anti-C1q antibody.
  • the present disclosure provides an anti-C1q antibody for use in treating or preventing Huntington's disease in an individual in need of such treatment.
  • the present disclosure provides use of an anti-C1q antibody in the manufacture of a medicament for treating or preventing Huntington's disease.
  • the present disclosure provides a kit comprising an anti-C1q antibody and a package insert comprising instructions for using the antibody to treat or prevent Huntington's disease in an individual in need of such treatment.
  • the anti-C1q antibody is: i) an isolated anti-C1q antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the light chain variable domain comprises the HVR-L1, HVR-L2, and HVR-L3 of the monoclonal antibody M1 produced by a hybridoma cell line with ATCC Accession Number PTA-120399 or progeny thereof; and/or wherein the heavy chain variable domain comprises the HVR-H1, HVR-H2, and HVR-H3 of the monoclonal antibody M1 produced by a hybridoma cell line with ATCC Accession Number PTA-120399 or progeny thereof; ii) an isolated anti-C1q antibody which binds essentially the same C1q epitope as the antibody M1 produced by the hybridoma cell line with ATCC Accession Number PTA-120399 or anti-C1q binding fragments thereof; or iii) an isolated murine anti-human C1
  • the anti-C1q antibody binds to a C1q protein and binds to one or more amino acids of the C1q protein within amino acid residues selected from the group consisting of: i) amino acid residues 196-226 of SEQ ID NO:1 (SEQ ID NO:6), or amino acid residues of a C1q protein chain A (C1qA) corresponding to amino acid residues 196-226 (GLFQVVSGGMVLQLQQGDQVWVEKDPKKGHI) of SEQ ID NO:1 (SEQ ID NO:6); ii) amino acid residues 196-221 of SEQ ID NO:1 (SEQ ID NO:7), or amino acid residues of a C1qA corresponding to amino acid residues 196-221 (GLFQVVSGGMVLQLQQGDQVWVEKDP) of SEQ ID.
  • amino acid residues 202-221 of SEQ ID NO:1 SEQ ID NO:8
  • amino acid residues of a C1qA corresponding to amino acid residues 202-221 SGGMVLQLQQGDQVWVEKD
  • amino acid residues 202-219 of SEQ ID NO:1 SEQ ID NO:9
  • amino acid residues of a C1qA corresponding to amino acid residues 202-219 SEQ ID NO:9
  • amino acid residues Lys 219 and/or Ser 202 of SEQ ID NO:1 amino acid residues of a C1qA corresponding Lys 219 and/or Ser 202 of SEQ ID NO:1.
  • the anti-C1q antibody binds to one or more amino acids of the C1q protein within amino acid residues selected from the group consisting of: i) amino acid residues 218-240 of SEQ ID NO:3 (SEQ ID NO:10) or amino acid residues of a C1q protein chain C (C1qC) corresponding to amino acid residues 218-240 (WLAVNDYYDMVGI QGSDSVFSGF) of SEQ ID NO:3 (SEQ ID NO:10); ii) amino acid residues 225-240 of SEQ ID NO:3 (SEQ ID NO:11) or amino acid residues of a C1qC corresponding to amino acid residues 225-240 (YDMVGI QGSDSVFSGF) of SEQ ID NO:3 (SEQ ID NO:11); iii) amino acid residues 225-232 of SEQ ID NO:3 (SEQ ID NO:12) or amino acid residues of a C1q protein within amino acid residues selected from the group consisting of: i
  • the anti-C1q antibody binds to a C1q protein and binds to one or more amino acids of the C1q protein chain A (C1qA) within amino acid residues selected from the group consisting of: i) amino acid residues 196-226 of SEQ ID NO:1 (SEQ ID NO:6), or amino acid residues of a C1q protein chain A (C1qA) corresponding to amino acid residues 196-226 (GLFQVVSGGMVLQLQQGDQVWVEKDPKKGHI) of SEQ ID NO:1 (SEQ ID NO:6); ii) amino acid residues 196-221 of SEQ ID NO:1 (SEQ ID NO:7), or amino acid residues of a C1qA corresponding to amino acid residues 196-221 (GLFQVVSGGMVLQLQQGDQVWVEKDP) of SEQ ID.
  • the anti-C1q antibody binds to amino acid residue Lys 219 and Ser 202 of the human C1qA as shown in SEQ ID NO:1 or amino acids of a human C1qA corresponding to Lys 219 and Ser 202 as shown in SEQ ID NO:1, and amino acid residue Tyr 225 of the human C1qC as shown in SEQ ID NO:3 or an amino acid residue of a human C1qC corresponding to Tyr 225 as shown in SEQ ID NO:3.
  • the anti-C1q antibody binds to amino acid residue Lys 219 of the human C1qA as shown in SEQ ID NO:1 or an amino acid residue of a human C1qA corresponding to Lys 219 as shown in SEQ ID NO:1, and amino acid residue Ser 185 of the human C1qC as shown in SEQ ID NO:3 or an amino acid residue of a human C1qC corresponding to Ser 185 as shown in SEQ ID NO:3.
  • the anti-C1q antibody binds specifically to both human C1q and mouse C1q. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody binds specifically to rat C1q. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody binds specifically to human C1q, mouse C1q, and rat C1q. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q has dissociation constant (K D ) for human C1q and mouse C1q that ranges from less than about 30 nM to less than about 100 pM.
  • K D dissociation constant
  • the anti-C1q antibody has dissociation constant (K D ) for human C1q and mouse C1q of less than about 30 nM. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody has dissociation constant (K D ) for human C1q and mouse C1q of less than about 20 nM. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody has dissociation constant (K D ) for human C1q and mouse C1q of less than about 10 nM.
  • the anti-C1q antibody has dissociation constant (K D ) for human C1q and mouse C1q of less than about 5 nM. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody has dissociation constant (K D ) for human C1q and mouse C1q of less than about 1 nM. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody has dissociation constant (K D ) for human C1q and mouse C1q of less than about 100 pM.
  • the anti-C1q antibody specifically binds to and neutralizes a biological activity of C1q.
  • the biological activity is (1) C1q binding to an autoantibody, (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 beta-amyloid, or (10) C1q binding to calreticulin.
  • the biological activity is (1) activation of the classical complement activation pathway, (2) activation of antibody and complement dependent cytotoxicity, (3) CH50 hemolysis, (4) synapse loss, (5) B-cell antibody production, (6) dendritic cell maturation, (7) T-cell proliferation, (8) cytokine production (9) microglia activation, (10) Arthus reaction, (11) phagocytosis of synapses or nerve endings, or (12) activation of complement receptor 3 (CR3/C3) expressing cells.
  • CH50 hemolysis comprises human, mouse, and/or rat CH50 hemolysis.
  • the anti-C1q antibody is capable of neutralizing at least 50%, at least 80%, or at least 90% of CH50 hemolysis. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody is capable of neutralizing at least 50% of CH50 hemolysis at a dose of less than 200 ng/ml, less than 100 ng/ml, less than 50 ng/ml, or less than 20 ng/ml. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody is a murine antibody. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody is a humanized, a chimeric antibody, or a human antibody.
  • the anti-C1q antibody is a multivalent antibody. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody is a bispecific antibody. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody has been engineered to increase brain penetration. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody is a bispecific antibody recognizing a first antigen and a second antigen. In certain embodiments that may be combined with any of the preceding embodiments, the first antigen is a C1q protein and the second antigen is an antigen facilitating transport across the blood-brain-barrier.
  • the second antigen is selected from the group consisting of transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor related proteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, CRM197, a llama single domain antibody, TMEM 30(A), a protein transduction domain, TAT, Syn-B, penetratin, a poly-arginine peptide, an angiopep peptide, and ANG1005.
  • the anti-C1q antibody is of the IgG class.
  • the anti-C1q antibody has an IgG 1 , IgG 2 , IgG 3 , or IgG 4 isotype. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody is an antibody fragment. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody is a Fab, F(ab′) 2 or Fab′ fragment. In certain embodiments that may be combined with any of the preceding embodiments, the antibody fragment specifically binds to and neutralizes a biological activity of C1q.
  • the antibody fragment has better brain penetration as compared to its corresponding full-length antibody. In certain embodiments that may be combined with any of the preceding embodiments, the antibody fragment has a shorter half-life as compared to its corresponding full-length antibody. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody inhibits C3 deposition. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody inhibits synapse loss.
  • the anti-C1q antibody inhibits complement-dependent cell-mediated cytotoxicity (CDCC) activation pathway by an amount that ranges from at least 30% to at least 99.9%. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody does not inhibit the lectin complement activation pathway. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody comprises a dissociation constant (K D ) for its corresponding antigen that ranges from 100 nM to 0.005 nM or less than 0.005 nM.
  • K D dissociation constant
  • the anti-C1q antibody inhibits autoantibody-dependent and complement-dependent cytotoxicity (CDC). In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody prevents amplification of the alternative complement activation pathway initiated by C1q binding. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody comprises an EC 50 that ranges from 3 ⁇ g/ml to 0.05 ⁇ g/ml, or less than 0.05 ⁇ g/ml. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody does not inhibit autoantibody-dependent cellular cytotoxicity (ADCC).
  • ADCC autoantibody-dependent cellular cytotoxicity
  • the method further comprises administering to the individual a therapeutically effective amount of a second anti-C1q antibody. In certain embodiments that may be combined with any of the preceding embodiments, the method further comprises administering to the individual a therapeutically effective amount of a second antibody, wherein the second antibody is selected from the group consisting of an anti-C1r antibody, an anti-C1s antibody and an anti-C1 complex antibody. In certain embodiments that may be combined with any of the preceding embodiments, the method further comprises administering to the individual a therapeutically effective amount of an inhibitor of antibody-dependent cellular cytotoxicity (ADCC).
  • ADCC antibody-dependent cellular cytotoxicity
  • the method further comprises administering to the individual a therapeutically effective amount of an inhibitor of the alternative complement activation pathway. In certain embodiments that may be combined with any of the preceding embodiments, the method further comprises administering to the individual an inhibitor of the interaction between an autoantibody and an autoantigen. In certain embodiments that may be combined with any of the preceding embodiments, the anti-C1q antibody binds a C1q antigen with a binding stoichiometry that ranges from 20:1 to 1.0:1 or less than 1.0:1.
  • the present disclosure provides for a diagnostic kit comprising an anti-C1q antibody of any of the preceding embodiments for treating or preventing Alzheimer's disease. In other aspects, the present disclosure provides for a diagnostic kit comprising an anti-C1q antibody of any of the preceding embodiments for treating or preventing Huntington's disease.
  • FIG. 1 illustrates the results of an ELISA screen for antibodies specifically binding human C1q.
  • Hybridoma supernatants containing anti-C1q antibodies 1C7, 2A1, 3A2, or 5A3 respectively were tested.
  • Left columns (grey) represent signals for anti-C1q antibody-binding to human C1q protein.
  • Right columns (black) represent signals for anti-C1q antibody-binding to human transferrin (HT).
  • FIG. 2 illustrates the C1q-neutralizing activities of anti-C1q antibodies 1C7, 2A1, 3A2, and 5A3 in a human CH50 hemolytic assay in a single-dose format.
  • FIG. 3 illustrates the C1q-neutralizing activities of anti-C1q antibodies 1C7, 3A2, and 4A4B 11 in a human CH50 hemolytic assay in a dose-response format.
  • FIG. 4 illustrates the C1q-neutralizing activities of anti-C1q antibodies M1 and 4A4B11 in human, mouse, and rat CH50 hemolytic assays in a dose-response format.
  • FIG. 4A illustrates results from a human CH50 hemolytic assay.
  • FIG. 4B illustrates results from a mouse CH50 hemolytic assay.
  • FIG. 4C illustrates results from a rat CH50 hemolytic assay.
  • FIG. 5 illustrates mass spectrometry characterization of C1q antibody complexes.
  • FIG. 5A shows a mixture of ANN-001 (4A4B 11) and C1q shows that ANN-001 monomer at the predicted mass of ⁇ 150 kDa, C1q monomer at the expected mass of ⁇ 460 kDa, and the C1q/ANN-001 1:1 complex at the predicted mass of ⁇ 600 kDa.
  • FIG. 5A shows a mixture of ANN-001 (4A4B 11) and C1q shows that ANN-001 monomer at the predicted mass of ⁇ 150 kDa, C1q monomer at the expected mass of ⁇ 460 kDa, and the C1q/ANN-001 1:1 complex at the predicted mass of ⁇ 600 kDa.
  • FIG. 5A shows a mixture of ANN-001 (4A4B 11) and C1q shows that ANN-001 monomer at the predicted mass of ⁇ 150 kDa, C1q monomer at the expected mass of
  • 5B shows a mixture of ANN-005 (M1) and C1q shows that ANN-005 monomer at the predicted mass of ⁇ 150 kDa, C1q monomer at the expected mass of ⁇ 460 kDa, and the C1q/ANN-005 1:1 complex at the predicted mass of ⁇ 600 kDa.
  • FIG. 6 illustrates C1q peptides do not compete with intact C1q for binding to monoclonal antibody ANN-005 (M1).
  • FIG. 6A depicts C1q and ANN-005 mixed in equimolar concentrations and incubated in the absence of a mixture of C1q peptides.
  • FIG. 6B depicts C1q and ANN-005 mixed in equimolar concentrations and incubated in the presence of a mixture of C1q peptides generated by pepsin digestion of C1q and analyzed by mass spectrometry.
  • ANN-005 and C1q a portion of the unbound antibody and antigen (ANN-005 and C1q) can be identified at the expected masses for monomers ( ⁇ 150 kDa and ⁇ 460kDa respectively) and a 1:1 complex is present at a mass of ⁇ 615 kDa.
  • FIG. 7A illustrates a general schematic representation of the complement cascade, including the three complement activation pathways and the terminal pathway.
  • FIG. 7B illustrated a schematic of the C1 complex. The C1s and C1r dimers are seen in a complex with the C1q hexamer.
  • FIG. 8 illustrates that A ⁇ oligomers induce C1q deposition and localization to synapses in wild-type brain. 18 hours post ICV injection of A ⁇ , brains were harvested for IHC.
  • FIG. 8A shows high-resolution images of C1q deposition in the hippocampus of wild-type (WT) mice. The left frame depicts C1q deposition in the hippocampus of control mice, middle frame depicts C1q deposition in the hippocampus of wild-type mice injected with A ⁇ monomers, and right frame depicts C1q deposition in the hippocampus of wild-type mice injected with A ⁇ oligomers.
  • WT wild-type mice
  • FIG. 9 illustrates how C1q deficiency suppresses the hippocampal synapse loss induced by soluble A ⁇ oligomers.
  • ICV injection of either soluble A ⁇ oligomers or A ⁇ monomers was performed in healthy adult (2-3 mo) wild-type (WT) mice. After 72 hours, brains were harvested for IHC using synapsin as a pre-synaptic marker and PSD95 as a post-synaptic marker.
  • FIG. 9A shows the labeling of synapses in control mice (left frame), mice injected with A ⁇ monomers (middle frame), and mice injected with A ⁇ oligomers (right frame).
  • FIG. 9A shows the labeling of synapses in control mice (left frame), mice injected with A ⁇ monomers (middle frame), and mice injected with A ⁇ oligomers (right frame).
  • FIG. 11 illustrates that anti-C1q antibodies can suppress complement deposition and synapse loss in an acute model of A ⁇ -induced synaptotoxicity.
  • FIG. 11A shows photomicrographs of C3 staining in the hippocampus of wild-type (WT) mice co-injected with A ⁇ oligomers and either a C1q blocking antibody or an IgG control (5 ng ICV plus 20 mg/kg IP).
  • FIG. 11B shows photomicrogaphs of PSD95 and Synapsin staining in the hippocampus of WT mice co-injected with A ⁇ oligomers and either a C1q blocking antibody or an IgG control.
  • FIG. 12 depicts fluorescence photomicrographs illustrating that the anti-C1q antibody M1 can prevent complement deposition in a transgenic mouse model of Huntington's disease (HD).
  • 20 mg/kg C1q blocking antibody and IgG control were injected IP (twice over a 48 hr period) into 5mo wild-type (WT) mice and zQ175 mice.
  • the term “preventing” includes providing prophylaxis with respect to occurrence or recurrence of a particular disease, disorder, or condition in an individual.
  • An individual may be predisposed to, susceptible to a particular disease, disorder, or condition, or at risk of developing such a disease, disorder, or condition, but has not yet been diagnosed with the disease, disorder, or condition.
  • an individual “at risk” of developing a particular disease, disorder, or condition may or may not have detectable disease or symptoms of disease, and may or may not have displayed detectable disease or symptoms of disease prior to the treatment methods described herein.
  • “At risk” denotes that an individual has one or more risk factors, which are measurable parameters that correlate with development of a particular disease, disorder, or condition, as known in the art. An individual having one or more of these risk factors has a higher probability of developing a particular disease, disorder, or condition than an individual without one or more of these risk factors.
  • treatment refers to clinical intervention designed to alter the natural course of the individual being treated during the course of clinical pathology. Desirable effects of treatment include decreasing the rate of progression, ameliorating or palliating the pathological state, and remission or improved prognosis of a particular disease, disorder, or condition.
  • An individual is successfully “treated”, for example, if one or more symptoms associated with a particular disease, disorder, or condition are mitigated or eliminated.
  • an “effective amount” refers to at least an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.
  • An effective amount can be provided in one or more administrations.
  • a “therapeutically effective amount” is at least the minimum concentration required to effect a measurable improvement of a particular disease, disorder, or condition.
  • a therapeutically effective amount herein may vary according to factors such as the disease state, age, sex, and weight of the patient, and the ability of the anti-C1q antibody to elicit a desired response in the individual.
  • a therapeutically effective amount may also be one in which any toxic or detrimental effects of the anti-C1q antibody are outweighed by the therapeutically beneficial effects.
  • Chronic administration refers to administration of the medicament(s) in a continuous as opposed to acute mode, so as to maintain the initial therapeutic effect (activity) for an extended period of time.
  • Intermittent administration refers to treatment that is not consecutively done without interruption, but rather is cyclic in nature.
  • administration “in conjunction” with another compound or composition includes simultaneous administration and/or administration at different times.
  • Administration in conjunction also encompasses administration as a co-formulation or administration as separate compositions, including at different dosing frequencies or intervals, and using the same route of administration or different routes of administration.
  • An “individual” for purposes of treatment, prevention, or reduction of risk refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sport, or pet animals, such as dogs, horses, rabbits, cattle, pigs, hamsters, gerbils, mice, ferrets, rats, cats, and the like. In some embodiments, the individual is human.
  • autoantibody means any antibody that recognizes a host antigen.
  • immunoglobulin (Ig) is used interchangeably with “antibody” herein.
  • antibody herein is used in the broadest sense and specifically covers monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g. bispecific antibodies) formed from at least two intact antibodies, and antibody fragments so long as they exhibit the desired biological activity.
  • the basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. The pairing of a V H and V L together forms a single antigen-binding site.
  • L light
  • H heavy
  • immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, having heavy chains designated alpha (“ ⁇ ”), delta (“ ⁇ ”), epsilon (“ ⁇ ”), gamma (“ ⁇ ”) and mu (“ ⁇ ”), respectively.
  • the ⁇ and ⁇ classes are further divided into subclasses (isotypes) on the basis of relatively minor differences in the CH sequence and function, e.g., humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.
  • subclasses immunoglobulins
  • the subunit structures and three dimensional configurations of different classes of immunoglobulins are well known and described generally in, for example, Abbas et al., Cellular and Molecular Immunology, 4 th ed. (W.B. Saunders Co., 2000).
  • “Native antibodies” are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (V H ) followed by a number of constant domains.
  • V H variable domain
  • Each light chain has a variable domain at one end (V L ) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light chain and heavy chain variable domains.
  • an “isolated” antibody such as an anti-C1q antibody of the present disclosure, is one that has been identified, separated and/or recovered from a component of its production environment (e.g., naturally or recombinantly).
  • the isolated polypeptide is free of association with all other contaminant components from its production environment.
  • Contaminant components from its production environment such as those resulting from recombinant transfected cells, are materials that would typically interfere with research, diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.
  • the polypeptide will be purified: (1) to greater than 95% by weight of antibody as determined by, for example, the Lowry method, and in some embodiments, to greater than 99% by weight; (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or silver stain.
  • Isolated antibody includes the antibody in situ within recombinant T-cells since at least one component of the antibody's natural environment will not be present. Ordinarily, however, an isolated polypeptide or antibody will be prepared by at least one purification step.
  • variable region refers to the amino-terminal domains of the heavy or light chain of the antibody.
  • the variable domains of the heavy chain and light chain may be referred to as “V H ” and “V L ”, respectively. These domains are generally the most variable parts of the antibody (relative to other antibodies of the same class) and contain the antigen binding sites.
  • variable refers to the fact that certain segments of the variable domains differ extensively in sequence among antibodies, such as anti-C1q antibodies of the present disclosure.
  • the V domain mediates antigen binding and defines the specificity of a particular antibody for its particular antigen.
  • HVRs hypervariable regions
  • FR framework regions
  • the variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three HVRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure.
  • the HVRs in each chain are held together in close proximity by the FR regions and, with the HVRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991)).
  • the constant domains are not involved directly in the binding of antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent-cellular toxicity.
  • the term “monoclonal antibody” as used herein refers to an antibody, such as an anti-C1q antibody of the present disclosure, obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and/or post-translation modifications (e.g., isomerizations, amidations) that may be present in minor amounts.
  • Monoclonal antibodies are highly specific, being directed against a single antigenic site. In contrast to polyclonal antibody preparations which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.
  • the monoclonal antibodies are advantageous in that they are synthesized by the hybridoma culture, uncontaminated by other immunoglobulins.
  • the modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • the monoclonal antibodies to be used in accordance with the present invention may be made by a variety of techniques, including, for example, the hybridoma method (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, 2d ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T - Cell Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No.
  • phage-display technologies see, e.g., Clackson et al., Nature, 352:624-628 (1991); Marks et 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. USA 101(34):12467-472 (2004); and Lee et al., J. Immunol.
  • full-length antibody “intact antibody” or “whole antibody” are used interchangeably to refer to an antibody, such as and anti-C1q antibody of the present disclosure, in its substantially intact form, as opposed to an antibody fragment.
  • whole antibodies include those with heavy and light chains including an Fc region.
  • the constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof.
  • the intact antibody may have one or more effector functions.
  • antibody fragment comprises a portion of an intact antibody, the antigen binding and/or the variable region of the intact antibody.
  • antibody fragments include Fab, Fab′, F(ab′) 2 and Fv fragments; diabodies; linear antibodies (see U.S. Pat. 5,641,870, Example 2; Zapata et al., Protein Eng. 8(10):1057-1062 (1995)); single-chain antibody molecules and multispecific antibodies formed from antibody fragments.
  • Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, and a residual “Fc” fragment, a designation reflecting the ability to crystallize readily.
  • the Fab fragment consists of an entire L chain along with the variable region domain of the H chain (V H ), and the first constant domain of one heavy chain (C H 1).
  • Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site.
  • Pepsin treatment of an antibody yields a single large F(ab′) 2 fragment which roughly corresponds to two disulfide linked Fab fragments having different antigen-binding activity and is still capable of cross-linking antigen.
  • Fab′ fragments differ from Fab fragments by having a few additional residues at the carboxy terminus of the C H 1 domain including one or more cysteines from the antibody hinge region.
  • Fab′-SH is the designation herein for Fab′ in which the cysteine residue(s) of the constant domains bear a free thiol group.
  • F(ab′) 2 antibody fragments originally were produced as pairs of Fab′ fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.
  • the Fc fragment comprises the carboxy-terminal portions of both H chains held together by disulfides.
  • the effector functions of antibodies are determined by sequences in the Fc region, the region which is also recognized by Fc receptors (FcR) found on certain types of cells.
  • “Fv” is the minimum antibody fragment which contains a complete antigen-recognition and -binding site. This fragment consists of a dimer of one heavy- and one light-chain variable region domain in tight, non-covalent association. From the folding of these two domains emanate six hypervariable loops (3 loops each from the H and L chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three HVRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.
  • Single-chain Fv also abbreviated as “sFv” or “scFv” are antibody fragments that comprise the VH and VL antibody domains connected into a single polypeptide chain.
  • the sFv polypeptide further comprises a polypeptide linker between the V H and V L domains which enables the sFv to form the desired structure for antigen binding.
  • Plückthun in The Pharmacology of Monoclonal Antibodies , vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).
  • “Functional fragments” of antibodies comprise a portion of an intact antibody, generally including the antigen binding or variable region of the intact antibody or the F region of an antibody which retains or has modified FcR binding capability.
  • antibody fragments include linear antibody, single-chain antibody molecules and multispecific antibodies formed from antibody fragments.
  • diabodies refers to small antibody fragments prepared by constructing sFv fragments (see preceding paragraph) with short linkers (about 5-10) residues) between the V H and V L domains such that inter-chain but not intra-chain pairing of the V domains is achieved, thereby resulting in a bivalent fragment, i.e., a fragment having two antigen-binding sites.
  • Bispecific diabodies are heterodimers of two “crossover” sFv fragments in which the V H and V L domains of the two antibodies are present on different polypeptide chains.
  • Diabodies are described in greater detail in, for example, EP 404,097; WO 93/11161; Hollinger et al., Proc. Nat'l Acad. Sci. USA 90:6444-48 (1993).
  • a “chimeric antibody” refers to an antibody (immunoglobulin), such as an anti-C1q antibody of the present disclosure, in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is(are) identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; Morrison et al., Proc. Nat'l Acad. Sci.
  • Chimeric antibodies of interest herein include PRIMATIZED® antibodies wherein the antigen-binding region of the antibody is derived from an antibody produced by, e.g., immunizing macaque monkeys with an antigen of interest.
  • PRIMATIZED® antibodies wherein the antigen-binding region of the antibody is derived from an antibody produced by, e.g., immunizing macaque monkeys with an antigen of interest.
  • humanized antibody is used a subset of “chimeric antibodies.”
  • “Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin.
  • a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from an HVR of the recipient are replaced by residues from an HVR of a non-human species (donor antibody) such as mouse, rat, rabbit or non-human primate having the desired specificity, affinity, and/or capacity.
  • donor antibody such as mouse, rat, rabbit or non-human primate having the desired specificity, affinity, and/or capacity.
  • FR residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody.
  • a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which 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 regions are those of a human immunoglobulin sequence, although the FR regions may include one or more individual FR residue substitutions that improve antibody performance, such as binding affinity, isomerization, immunogenicity, and the like.
  • the number of these amino acid substitutions in the FR is typically no more than 6 in the H chain, and in the L chain, no more than 3.
  • the humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • a “human antibody” is one that possesses an amino-acid sequence corresponding to that of an antibody, such as an anti-C1q antibody of the present disclosure, produced by a human and/or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising 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).
  • Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e.g., immunized xenomice (see, e.g., U.S. Pat. Nos.
  • hypervariable region when used herein refers to the regions of an antibody-variable domain, such as that of an anti-C1q antibody of the present disclosure, that are hypervariable in sequence and/or form structurally defined loops.
  • antibodies comprise six HVRs; three in the VH (H1, H2, H3), and three in the VL (L1, L2, L3).
  • H3 and L3 display the most diversity of the six HVRs, and H3 in particular is believed to play a unique role in conferring fine specificity to antibodies.
  • HVR delineations are in use and are encompassed herein.
  • the HVRs that are Kabat complementarity-determining regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., supra). Chothia refers instead to the location of the structural loops (Chothia and Lesk. J. Mol. Biol. 196:901-917 (1987)).
  • the AbM HVRs represent a compromise between the Kabat CDRs and Chothia structural loops, and are used by Oxford Molecular's AbM antibody-modeling software.
  • the “contact” HVRs are based on an analysis of the available complex crystal structures. The residues from each of these HVRs are noted below.
  • HVRs may comprise “extended HVRs” as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in the VL, and 26-35 (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in the VH.
  • the variable-domain residues are numbered according to Kabat et al., supra, for each of these extended-HVR definitions.
  • Framework or “FR” residues are those variable-domain residues other than the HVR residues as herein defined.
  • variable-domain residue-numbering as in Kabat or “amino-acid-position numbering as in Kabat,” and variations thereof, refers to the numbering system used for heavy-chain variable domains or light-chain variable domains of the compilation of antibodies in Kabat et al., supra. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion into, a FR or HVR of the variable domain.
  • a heavy-chain variable domain may include a single amino acid insert (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g., residues 82a, 82b, and 82c, etc. according to Kabat) after heavy-chain FR residue 82.
  • the Kabat numbering of residues may be determined for a given antibody by alignment at regions of homology of the sequence of the antibody with a “standard” Kabat numbered sequence.
  • the Kabat numbering system is generally used when referring to a residue in the variable domain (approximately 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 a residue in an immunoglobulin heavy chain constant region (e.g., the EU index reported in Kabat et al., supra).
  • the “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.
  • references to residue numbers in the variable domain of antibodies means residue numbering by the Kabat numbering system. Unless stated otherwise herein, references to residue numbers in the constant domain of antibodies means residue numbering by the EU numbering system (e.g., see United States Patent Publication No. 2010-280227).
  • acceptor human framework is a framework comprising the amino acid sequence of a VL or VH framework derived from a human immunoglobulin framework or a human consensus framework.
  • An acceptor human framework “derived from” a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence thereof, or it may contain pre-existing amino acid sequence changes. In some embodiments, the number of pre-existing amino acid changes are 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.
  • VL acceptor human framework is identical in sequence to the VL human immunoglobulin framework sequence or human consensus framework sequence.
  • a “human consensus framework” is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences.
  • the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences.
  • the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Examples include for the VL, the subgroup may be subgroup kappa I, kappa II, kappa III or kappa IV as in Kabat et al., supra. Additionally, for the VH, the subgroup may be subgroup I, subgroup II, or subgroup III as in Kabat et al., supra.
  • amino-acid modification at a specified position, e.g., of an anti-C1q antibody of the present disclosure, refers to the substitution or deletion of the specified residue, or the insertion of at least one amino acid residue adjacent the specified residue. Insertion “adjacent” to a specified residue means insertion within one to two residues thereof. The insertion may be N-terminal or C-terminal to the specified residue.
  • the amino acid modification herein is a substitution.
  • an “affinity-matured” antibody such as an anti-C1q antibody of the present disclosure, is one with one or more alterations in one or more HVRs thereof that result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody that does not possess those alteration(s).
  • an affinity-matured antibody has nanomolar or even picomolar affinities 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) describes affinity maturation by VH- and VL-domain shuffling. Random mutagenesis of HVR and/or framework residues is described by, for example: Barbas et al.
  • the term “specifically recognizes” or “specifically binds” refers to measurable and reproducible interactions such as attraction or binding between a target and an antibody, such as an anti-C1q antibody of the present disclosure, that is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules.
  • an antibody, such as an anti-C1q antibody of the present disclosure, that specifically or preferentially binds to a target or an epitope is an antibody that binds this target or epitope with greater affinity, avidity, more readily, and/or with greater duration than it binds to other targets or other epitopes of the target.
  • an antibody (or a moiety) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target.
  • “specific binding” or “preferential binding” does not necessarily require (although it can include) exclusive binding.
  • An antibody that specifically binds to a target may have an association constant of 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 instances about 10 6 M ⁇ 1 or 10 7 M ⁇ 1 , about 10 8 M ⁇ 1 to 10 9 M ⁇ 1 , or about 10 10 M to 10 11 M ⁇ 1 or higher.
  • immunoassay formats can be used to select antibodies specifically immunoreactive with a particular protein.
  • solid-phase ELISA immunoassays are routinely used to select monoclonal antibodies specifically immunoreactive with a protein. See, e.g., Harlow and Lane (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity.
  • an “interaction” between a complement protein, such as complement factor C1 q, and a second protein encompasses, without limitation, protein-protein interaction, a physical interaction, a chemical interaction, binding, covalent binding, and ionic binding.
  • an antibody “inhibits interaction” between two proteins when the antibody disrupts, reduces, or completely eliminates an interaction between the two proteins.
  • blocking antibody, an “antagonist” antibody, an “inhibitory” antibody, or a “neutralizing” antibody is an antibody, such as an anti-C1q antibody of the present disclosure that inhibits or reduces one or more biological activities of the antigen it binds, such as interactions with one or more proteins.
  • blocking antibodies, antagonist antibodies, inhibitory antibodies, or “neutralizing” antibodies substantially or completely inhibit one or more biological activities or interactions of the antigen.
  • Antibody effector functions refer to those biological activities attributable to the Fc region (a native sequence Fc region or amino acid sequence variant Fc region) of an antibody, and vary with the antibody isotype.
  • Fc region herein is used to define a C-terminal region of an immunoglobulin heavy chain, including native-sequence Fc regions and variant Fc regions.
  • the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy-chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof.
  • the C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during production or purification of the antibody, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody.
  • composition of intact antibodies may comprise antibody populations with all K447 residues removed, antibody populations with no K447 residues removed, and antibody populations having a mixture of antibodies with and without the K447 residue.
  • Suitable native-sequence Fc regions for use in the antibodies of the invention include human IgG1, IgG2, IgG3 and IgG4.
  • a “native sequence Fc region” comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature.
  • Native sequence human Fc regions include a native sequence human IgG1 Fc region (non-A and A allotypes); native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc region as well as naturally occurring variants thereof.
  • a “variant Fc region” comprises an amino acid sequence which differs from that of a native sequence Fc region by virtue of at least one amino acid modification. In some embodiments, the variant Fc region differs in one or more amino acid substitution(s). In some embodiments, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or to the Fc region of a parent polypeptide, e.g. from about one to about ten amino acid substitutions, and, in some embodiments, from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide.
  • the variant Fc region herein will, in some embodiments, possess at least about 80% homology with a native sequence Fc region and/or with an Fc region of a parent polypeptide, and, in some embodiments, at least about 90% homology therewith, and, in some embodiments, at least about 95% homology therewith.
  • Fc receptor or “FcR” describes a receptor that binds to the Fc region of an antibody.
  • the FcR is a native sequence human FcR.
  • a FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the Fc ⁇ RI, Fc ⁇ RII, and FcyR ⁇ III subclasses, including allelic variants and alternatively spliced forms of these receptors, Fc ⁇ RII receptors include Fc ⁇ RIIA (an “activating receptor”) and Fc ⁇ RIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof.
  • Activating receptor Fc ⁇ RIIA contains an immunoreceptor tyrosine-based activation motif (“ITAM”) in its cytoplasmic domain.
  • Inhibiting receptor Fc ⁇ RIIB contains an immunoreceptor tyrosine-based inhibition motif (“ITIM”) in its cytoplasmic domain.
  • ITAM immunoreceptor tyrosine-based activation motif
  • ITIM immunoreceptor tyrosine-based inhibition motif
  • Binding to FcRn in vivo and serum half-life of human FcRn high-affinity binding polypeptides can be assayed, e.g., in transgenic mice or transfected human cell lines expressing human FcRn, or in primates to which the polypeptides having a variant Fc region are administered.
  • WO 2004/42072 (Presta) describes antibody variants with improved or diminished binding to FcRs. See also, e.g., Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001).
  • k on is intended to refer to the rate constant for association of an antibody to an antigen.
  • k off is intended to refer to the rate constant for dissociation of an antibody from the antibody/antigen complex.
  • K D is intended to refer to the equilibrium dissociation constant of an antibody-antigen interaction.
  • percent (%) amino acid sequence identity and “homology” with respect to a peptide, polypeptide or antibody sequence refers to the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the specific peptide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALIGNTM (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms known in the art needed to achieve maximal alignment over the full length of the sequences being compared.
  • an “isolated” molecule or cell is a molecule or a cell that is identified and separated from at least one contaminant molecule or cell with which it is ordinarily associated in the environment in which it was produced.
  • the isolated molecule or cell is free of association with all components associated with the production environment.
  • the isolated molecule or cell is in a form other than in the form or setting in which it is found in nature. Isolated molecules therefore are distinguished from molecules existing naturally in cells; isolated cells are distinguished from cells existing naturally in tissues, organs, or individuals.
  • the isolated molecule is an anti-C1q antibody of the present disclosure.
  • the isolated cell is a host cell or hybridoma cell producing an anti-C1q antibody of the present disclosure.
  • an “isolated” nucleic acid molecule encoding an antibody is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the environment in which it was produced. In some embodiments, the isolated nucleic acid is free of association with all components associated with the production environment.
  • the isolated nucleic acid molecules encoding the polypeptides and antibodies herein is in a form other than in the form or setting in which it is found in nature. Isolated nucleic acid molecules therefore are distinguished from nucleic acid encoding the polypeptides and antibodies herein existing naturally in cells.
  • vector is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
  • plasmid refers to a circular double stranded DNA into which additional DNA segments may be ligated.
  • phage vector refers to a viral vector, wherein additional DNA segments may be ligated into the viral genome.
  • viral vector capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors).
  • vectors e.g., non-episomal mammalian vectors
  • vectors can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome.
  • certain vectors are capable of directing the expression of genes to which they are operatively linked.
  • Such vectors are referred to herein as “recombinant expression vectors,” or simply, “expression vectors.”
  • expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
  • plasmid and “vector” may be used interchangeably as the plasmid is the most commonly used form of vector.
  • Polynucleotide or “nucleic acid,” as used interchangeably herein, refer to polymers of nucleotides of any length, and include DNA and RNA.
  • the nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction.
  • a polynucleotide may comprise modified nucleotides, such as methylated nucleotides and their analogs. If present, modification to the nucleotide structure may be imparted before or after assembly of the polymer.
  • the sequence of nucleotides may be interrupted by non-nucleotide components.
  • a polynucleotide may comprise modification(s) made after synthesis, such as conjugation to a label.
  • modifications include, for example, “caps,” substitution of one or more of the naturally occurring nucleotides with an analog, internucleotide modifications such as, for example, those with uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoamidates, carbamates, etc.) and with charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), those containing pendant moieties, such as, for example, proteins (e.g., nucleases, toxins, antibodies, signal peptides, ply-L-lysine, etc.), those with intercalators (e.g., acridine, psoralen, etc.), those containing chelators (e.g., metals,
  • any of the hydroxyl groups ordinarily present in the sugars may be replaced, for example, by phosphonate groups, phosphate groups, protected by standard protecting groups, or activated to prepare additional linkages to additional nucleotides, or may be conjugated to solid or semi-solid supports.
  • the 5′ and 3′ terminal OH can be phosphorylated or substituted with amines or organic capping group moieties of from 1 to 20 carbon atoms.
  • Other hydroxyls may also be derivatized to standard protecting groups.
  • Polynucleotides can also contain analogous forms of ribose or deoxyribose sugars that are generally known in the art, including, for example, 2′-O-methyl-, 2′-O-allyl-, 2′-fluoro- or 2′-azido-ribose, carbocyclic sugar analogs, ⁇ -anomeric sugars, epimeric sugars such as arabinose, xyloses or lyxoses, pyranose sugars, furanose sugars, sedoheptuloses, acyclic analogs, and basic nucleoside analogs such as methyl riboside.
  • One or more phosphodiester linkages may be replaced by alternative linking groups.
  • linking groups include, but are not limited to, embodiments wherein 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”), in which each R or R′ is independently H or substituted or unsubstituted alkyl (1-20 C) optionally containing an ether (—O—) linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl or araldyl. Not all linkages in a polynucleotide need be identical. The preceding description applies to all polynucleotides referred to herein, including RNA and DNA.
  • a “host cell” includes an individual cell or cell culture that can be or has been a recipient for vector(s) for incorporation of polynucleotide inserts.
  • Host cells include progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or in genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation.
  • a host cell includes cells transfected in vivo with a polynucleotide(s) of this invention.
  • Carriers as used herein include pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffered solution.
  • physiologically acceptable carriers include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or nonionic surfactants such as TWEENTM, polyethylene glycol (PEG), and PLURONIC STM.
  • buffers such as phosphate, citrate, and other organic acids
  • antioxidants including ascorbic acid
  • proteins such as serum albumin
  • an “antibody” is a reference to from one to many antibodies, such as molar amounts, and includes equivalents thereof known to those skilled in the art, and so forth.
  • the present disclosure provides methods for treating, preventing, or reducing risk of Alzheimer's disease and/or Huntington's disease.
  • inhibition of the classical pathway of complement activation is an effective therapeutic strategy for the treatment of Alzheimer's disease and Hntington's disease (FIG. 7A).
  • effective strategies for inhibiting the classical pathway include inhibiting the interaction between C1q and autoantibodies, such as anti-ganglioside autoantibodies, inhibiting the interaction between C1q and C1r or C1s, blocking the catalytic activity of C1r or C1s, and blocking the interactions between C1r or C1s and their respective substrates ( FIG. 7A ).
  • an autoantibody of the present disclosure includes, without limitation, an antibody that recognizes a host antigen and activates the classical pathway of complement activation. In the first step of this activation process complement factor C1q binds to the autoantibody-autoantigen-immune complex.
  • certain aspects of the present disclosure relate to anti-C1q antibodies for use in treating, preventing, or reducing risk of Alzheimer's disease and/or Huntinton disease, in individuals in need thereof.
  • the present disclosure provides methods for treating or preventing Alzheimer s disease and/or Huntington's disease, in an individual in need of such treatment, by administering to the individual a therapeutically effective dose of an anti-C1q antibody.
  • the anti-C1q antibody is a C1q neutralizing antibody.
  • the anti-C1q antibody binds to C1 complex.
  • the anti-C1q antibody inhibits the interaction between C1q and an autoantibody, between C1q and C1r, and/or between C1q and C1s.
  • the individual has Alzheimer's disease or Huntington's disease.
  • the individual is a human.
  • anti-C1q antibodies and uses thereof.
  • the anti-C1q antibodies of this disclosure specifically bind a C1q protein of this disclosure.
  • the anti-C1q antibodies are C1q neutralizing antibodies.
  • the anti-C1q antibodies of this disclosure may bind to C1 complex.
  • the present disclosure provides murine monoclonal antibody M1, which is produced by a hybridoma cell line referred to as mouse hybridoma C1q-M1 7788-1(M) 051613 and which was deposited with ATCC on Jun. 6, 2013 with ATCC Accession Number PTA-120399.
  • the present disclosure provides an anti-C1q antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the light chain variable domain comprises the light chain variable domain sequence of antibody M1; and/or wherein the heavy chain comprises the heavy chain variable domain sequence of antibody M1.
  • the present disclosure provides an anti-C1q antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the light chain variable domain comprises the HVR-L1, HVR-L2, and HVR-L3 of monoclonal antibody M1 produced by a hybridoma cell line deposited at ATCC with ATCC Accession Number PTA-120399 or progeny thereof; and/or wherein the heavy chain variable domain comprises the HVR-H1, HVR-H2, and HVR-H3 of monoclonal antibody M1 produced by a hybridoma cell line deposited at ATCC with Accession Number PTA-120399 or progeny thereof.
  • the present disclosure provides an anti-C1q antibody, which binds essentially the same C1q epitope as (1) antibody M1 produced by the hybridoma cell line deposited with ATCC on Jun. 6, 2013 and having ATCC Accession Number PTA-120399 or progeny thereof, (2) an antigen binding fragment of antibody Ml, or (3) an antibody comprising the HVR-L1, HVR-L2, HVR-L3, HVR-H1, HVR-H2, and HVR-H3 of antibody M1.
  • the anti-C1q antibodies of this disclosure neutralize a biological activity of C1q.
  • Uses for anti-C1q antibodies include, without limitation, the detection of complement factor C1q, e.g., in individuals having a neurodegenerative disorder associated with complement factor 1 (CF 1)-dependent pathological synapse loss, such as Alzheimer's disease and/or Huntington's disease.
  • Additional non-limiting uses include the inhibition of the classical pathway of complement activation, e.g., in cases where the classical complement pathway is activated by autoantibodies, such as ganglioside-specific autoantibodies.
  • anti-C1q antibodies include the diagnosis and treatment of disorders that are associated with elevated expression of complement factors, such as C1q, or associated with the activation of the complement pathway.
  • disorders may include, without limitation, autoimmune disorders, inflammatory disorders, and neurodegenerative disorders, including neurodegenerative disorders associated with synapse loss such as Alzheimer's disease and Huntington's disease.
  • the present disclosure provides an isolated nucleic acid molecule encoding an antibody of this disclosure.
  • the present disclosure also provides isolated host cells containing a nucleic acid molecule that encodes an antibody of this disclosure.
  • an isolated host cell line is provided that produces the neutralizing monoclonal murine antibody M1. This isolated host cell lines was deposited with ATCC and has ATCC Accession Number PTA-120399.
  • compositions are provided containing anti-C1q antibodies of this disclosure, such as C1q neutralizing antibodies of this disclosure, in combination with pharmaceutically acceptable carriers.
  • the present disclosure also provides a kit containing an anti-C1q antibody for use in any of the methods described herein.
  • the present disclosure further provides methods of using the anti-C1q antibodies of this disclosure (e.g., C1q neutralizing antibodies of this disclosure) to treat or prevent a neurodegenerative disease (e.g., Alzheimer's disease and Huntington's disease) in an individual in need of such treatment, to detect synapses in an individual having a neurodegenerative disease, and to detect synapses in a biological sample.
  • a neurodegenerative disease e.g., Alzheimer's disease and Huntington's disease
  • kits containing the C1q antibodies of this disclosure e.g., C1q neutralizing antibodies of this disclosure.
  • the methods of this disclosure involve administering or using antibodies that specifically recognizes complement factor C1q of the classical complement activation pathway.
  • Certain aspects of the present disclosure further involves antibodies that specifically recognize complement factor C1q and/or C1q in the C1 complex of the classical complement activation pathway.
  • the recognized complement factor may be derived, without limitation, from any organism having a complement system, including any mammalian organism such as human, mouse, rat, rabbit, monkey, dog, cat, cow, horse, camel, sheep, goat, or pig.
  • C1 complex refers to a protein complex that may include, without limitation, one C1q protein, two C1r proteins, and two C1s proteins (e.g., C1qr 2 s 2 ).
  • complement factor C1q refers to both wild type sequences and naturally occurring variant sequences.
  • a non-limiting example of a complement factor C1q recognized by antibodies of this invention is human C1q, including the three polypeptide chains A, B, and C:
  • Protein Data Base NP_000482.3; GenBank No.: NM_000491.3: >gi
  • Protein Data Base NP_001107573.1; GenBank No.: NM_001114101.1: >gi
  • an anti-C1q antibody of the present disclosure may bind to polypeptide chain A, polypeptide chain B, and/or polypeptide chain C of a C1q protein.
  • an anti-C1q antibody of the present disclosure binds to polypeptide chain A, polypeptide chain B, and/or polypeptide chain C of human C1q or a homolog thereof, such as mouse, rat, rabbit, monkey, dog, cat, cow, horse, camel, sheep, goat, or pig C1q.
  • the antibodies of this disclosure specifically bind to a complement factor C1q and/or C1q in the C1 complex of the classical complement pathway.
  • the anti-C1q antibodies specifically bind to human C1q.
  • the anti-C1q antibodies specifically bind to human and mouse C1q.
  • the anti-C1q antibodies specifically bind to rat C1q.
  • the anti-C1q antibodies specifically bind to human C1q, mouse C1q, and rat C1q.
  • the anti-C1q antibodies of this disclosure neutralize a biological activity of complement factor C1q.
  • the antibodies inhibit the interaction between complement factor C1q and other complement factors, such as C1r or C1s or between C1q and an antibody, such as an autoantibody.
  • the antibodies inhibit the interaction between complement factor C1q and a non-complement factor.
  • a non-complement factor may include phosphatidylserine, pentraxin-3, C-reactive protein (CRP), globular C1q receptor (gC1qR), complement receptor 1 (CR1), ⁇ -amyloid, and calreticulin.
  • the antibodies inhibit the classical complement activation pathway.
  • the antibodies further inhibit the alternative pathway.
  • the antibodies inhibit autoantibody- and complement-dependent cytotoxicity (CDC).
  • the antibodies inhibit complement-dependent cell-mediated cytotoxicity (CDCC).
  • the antibodies inhibit B-cell antibody production, dendritic cell maturation, T-cell proliferation, cytokine production, or microglia activation.
  • the antibodies inhibit the Arthus reaction.
  • the antibodies inhibit phagocytosis of synapses or nerve endings.
  • the antibodies inhibit the activation of complement receptor 3 (CR3/C3) expressing cells.
  • C3/C3 complement receptor 3
  • the functional properties of the antibodies of this invention may, without limitation, be measured in in vitro, ex vivo, or in vivo experiments.
  • the dissociation constants (K D ) of the anti-C1q antibodies for C1q may be less than 100 nM, less than 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 9 nM, less than 8 nM, less than 7 nM, less than 6 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, less than 0.05 nM, less than 0.01 nM, or less than 0.005 nM.
  • dissociation constants range from less than about 30 nM to less than about 100 pM. In some embodiments, dissociation constants are less than about 30 nM. In some embodiments, dissociation constants are less than about 20 nM. In some embodiments, dissociation constants are less than about 10 nM. In some embodiments, dissociation constants are less than about 5 nM. In some embodiments, dissociation constants are less than about 1 nM. In some embodiments, dissociation constants are less than about 100 pM.
  • the dissociation constants of the anti-C1q antibody range from less than about 30 nM to less than about 100 pM for human C1q, and range from less than about 30 nM to less than about 100 pM for mouse C1q. In certain embodiments, dissociation constants of the anti-C1q antibody are less than about 30 nM for human C1q and less than about 30 nM for mouse C1q. In certain embodiments, dissociation constants of the anti-C1q antibody are less than about 20 nM for human C1q and less than about 20 nM for mouse C1q.
  • dissociation constants of the anti-C1q antibody are less than about 10 nM for human C1q and less than about 10 nM for mouse C1q. In certain embodiments, dissociation constants of the anti-C1q antibody are less than about 5 nM for human C1q and less than about 5 nM for mouse C1q. In certain embodiments, dissociation constants of the anti-C1q antibody are less than about 1 nM for human C1q and less than about 1 nM mouse C1q. In certain embodiments, the dissociation constants of the anti-C1q antibody are less than 100 pM for human C1q and less than 100 pM for mouse C1q.
  • Antibody dissociation constants for antigens other than C1q may be least 5-fold, at least 10-fold, at least 100-fold, at least 1,000-fold, at least 10,000-fold, or at least 100,000-fold higher that the dissociation constants for C1q.
  • the dissociation constant of a C1q antibody of this disclosure may be at least 1,000-fold higher for C1s than for C1q.
  • Dissociation constants may be determined through any analytical technique, including any biochemical or biophysical technique such as ELISA, surface plasmon resonance (SPR), bio-layer interferometry (see, e.g., Octet System by ForteBio), isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC), circular dichroism (CD), stopped-flow analysis, and colorimetric or fluorescent protein melting analyses.
  • Dissociation constants (K D ) of the anti-C1q antibodies for C1q may be determined, e.g., using full-length antibodies or antibody fragments, such as Fab fragments.
  • One exemplary way of determining binding affinity of antibodies to C1q is by measuring binding affinity of monofunctional Fab fragments of the antibody.
  • an antibody for example, IgG
  • an antibody can be cleaved with papain or expressed recombinantly.
  • the affinity of an Fab fragment of an antibody can be determined by surface plasmon resonance (Biacore3000TM surface plasmon resonance (SPR) system, BiacoreTM, INC, Piscataway N.J.) equipped with pre-immobilized streptavidin sensor chips (SA) using HBS-EP running buffer (0.01M HEPES, pH 7.4, 0.15 NaCl, 3mM EDTA, 0.005% v/v Surfactant P20).
  • Biotinylated human C1q (or any other C1q) can be diluted into HBS-EP buffer to a concentration of less than 0.5 ⁇ g/mL and injected across the individual chip channels using variable contact times, to achieve two ranges of antigen density, either 50-200 response units (RU) for detailed kinetic studies or 800-1,000 RU for screening assays. Regeneration studies have shown that 25 mM NaOH in 25% v/v ethanol effectively removes the bound Fab while keeping the activity of C1q on the chip for over 200 injections. Typically, serial dilutions (spanning concentrations of 0.1-10.times.
  • K D estimated K D of purified Fab samples are injected for 1 min at 100 ⁇ L/minute and dissociation times of up to 2 hours are allowed.
  • concentrations of the Fab proteins are determined by ELISA and/or SDS-PAGE electrophoresis using a Fab of known concentration (as determined by amino acid analysis) as a standard.
  • Kinetic association rates (k on ) and dissociation rates (k off ) are obtained simultaneously by fitting the data globally to a 1:1 Langmuir binding model (Karlsson, R. Roos, H. Fagerstam, L. Petersson, B. (1994). Methods Enzymology 6. 99-110) using the BIAevaluation program.
  • Equilibrium dissociation constant (K D ) values are calculated as k off /k on . This protocol is suitable for use in determining binding affinity of an antibody to any C1q, including human C1q, C1q of another mammal (such as mouse C1q, rat C1q, primate C1q), as well as different forms of C1q. Binding affinity of an antibody is generally measured at 25° C., but can also be measured at 37° C.
  • the antibodies of this disclosure may bind to C1q antigens derived from any organism having a complement system, including any mammalian organism such as human, mouse, rat, rabbit, monkey, dog, cat, cow, horse, camel, sheep, goat, or pig.
  • the anti-C1q antibodies bind specifically to epitopes on human C1q.
  • the anti-C1q antibodies specifically bind to epitopes on both human and mouse C1q.
  • the anti-C1q antibodies specifically bind to epitopes on human, mouse, and rat C1q.
  • an anti-C1q antibody that binds to an epitope of C1q that is the same as or overlaps with the C1q epitope bound by another antibody of this disclosure.
  • an anti-C1q antibody that binds to an epitope of C1q that is the same as or overlaps with the C1q epitope bound by anti-C1q antibody Ml.
  • the anti-C1q antibody competes with another antibody of this disclosure for binding to C1q.
  • the anti-C1q antibody competes with anti-C1q antibody M1 or an antigen-binding fragment thereof for binding to C1q.
  • Methods that may be used to determine which C1q epitope of an anti-C1q antibody binds to, or whether two antibodies bind to the same or an overlapping epitope may include, without limitation, X-ray crystallography, NMR spectroscopy, Alanine-Scanning Mutagenesis, the screening of peptide libraries that include C1q-derived peptides with overlapping C1q sequences, and competition assays.
  • Competition assays are especially useful to determine whether two antibodies bind the same epitope by recognizing identical or sterically overlapping epitopes or whether one antibody competitively inhibits binding of another antibody to the antigen. These assays are known in the art.
  • an antigen or antigen expressing cells are immobilized on a multi-well plate and the ability of unlabeled antibodies to block the binding of labeled antibodies is measured.
  • Common labels for such competition assays are radioactive labels or enzyme labels.
  • competitive antibodies encompassed herein are antibodies that inhibit (i.e., prevent or interfere with in comparison to a control) or reduce the binding of any anti-C1q antibody of this disclosure (such as M1 or an antigen-binding fragment of M1) to C1q by at least 50%, 60%, 70%, 80%, 90% and 95% at 1 ⁇ M or less.
  • concentration competing antibody in the competition assay may be at or below the K D of antibody M1 or an antigen-binding fragment of Ml. Competition between binding members may be readily assayed in vitro for example using ELISA and/or by monitoring the interaction of the antibodies with C1q in solution. The exact means for conducting the analysis is not critical.
  • C1q may be immobilized to a 96-well plate or may be placed in a homogenous solution.
  • the ability of unlabeled candidate antibody(ies) to block the binding of the labeled anti-C1q antibody, e.g. M1 can be measured using radioactive, enzyme or other labels.
  • the ability of unlabeled antibodies to interfere with the interaction of a labeled anti-C1q antibody with C1q wherein said labeled anti-C1q antibody, e.g., M1, and C1q are already bound is determined. The readout is through measurement of bound label.
  • C1q and the candidate antibody(ies) may be added in any order or at the same time.
  • the anti-C1q antibody inhibits the interaction between C1q and an autoantibody.
  • the anti-C1q antibody is murine anti-human C1q monoclonal antibody Ml, which is produced by a hybridoma cell line deposited with ATCC on June 6, 2013 with ATCC Accession Number PTA-120399.
  • the anti-C1q antibody is an isolated antibody which binds essentially the same C1q epitope as M1.
  • the anti-C1q antibody is an isolated antibody comprising the HVR-L1, HVR-L2, and HVR-L3 of the light chain variable domains of monoclonal antibody M1 produced by the hybridoma cell line deposited with ATCC on Jun. 6, 2013 with ATCC Accession Number PTA-120399, or progeny thereof.
  • the anti-C1q antibody is an isolated antibody comprising the HVR-H1, HVR-H2, and HVR-H3 of the heavy chain variable domains of monoclonal antibody M1 produced by the hybridoma cell line deposited with ATCC on Jun.
  • the anti-C1q antibody is an isolated antibody comprising the HVR-L1, HVR-L2, and HVR-L3 of the light chain variable domains and the HVR-H1, HVR-H2, and HVR-H3 of the heavy chain variable domains of monoclonal antibody M1 produced by the hybridoma cell line deposited with ATCC on Jun. 6, 2013 with ATCC Accession Number PTA-120399, or progeny thereof.
  • the anti-C1q antibody binds to a C1q protein and binds to one or more amino acids of the C1q protein within amino acid residues selected from (a) amino acid residues 196-226 of SEQ ID NO:1 (SEQ ID NO:6), or amino acid residues of a C1q protein chain A (C1qA) corresponding to amino acid residues 196-226 (GLFQVVSGGMVLQLQQGDQVWVEKDPKKGHI) of SEQ ID NO:1 (SEQ ID NO:6); (b) amino acid residues 196-221 of SEQ ID NO:1 (SEQ ID NO:7), or amino acid residues of a C1qA corresponding to amino acid residues 196-221 (GLFQVVSGGMVLQLQQGDQVWVEKDP) of SEQ ID.
  • amino acid residues 202-221 of SEQ ID NO:1 SEQ ID NO:8
  • amino acid residues 202-219 of SEQ ID NO:1 SEQ ID NO:9
  • amino acid residues of a C1qA corresponding to amino acid residues 202-219 SEQ ID NO:9
  • amino acid residues Lys 219 and/or Ser 202 of SEQ ID NO:1 amino acid residues of a C1qA corresponding Lys 219 and/or Ser 202 of SEQ ID NO:1.
  • the antibody further binds to one or more amino acids of the C1q protein within amino acid residues selected from: (a) amino acid residues 218-240 of SEQ ID NO:3 (SEQ ID NO:10) or amino acid residues of a C1q protein chain C (C1qC) corresponding to amino acid residues 218-240 (WLAVNDYYDMVGI QGSDSVFSGF) of SEQ ID NO:3 (SEQ ID NO:10); (b) amino acid residues 225-240 of SEQ ID NO:3 (SEQ ID NO:11) or amino acid residues of a C1qC corresponding to amino acid residues 225-240 (YDMVGI QGSDSVFSGF) of SEQ ID NO:3 (SEQ ID NO:11); (c) amino acid residues 225-232 of SEQ ID NO:3 (SEQ ID NO:12) or amino acid residues of a C1qC corresponding to amino acid residues 225-232 (YDMVGIQG) of SEQ ID NO
  • the anti-C1q antibody binds to amino acid residue Lys 219 and Ser 202 of the human C1qA as shown in SEQ ID NO:1 or amino acids of a human C1qA corresponding to Lys 219 and Ser 202 as shown in SEQ ID NO:1, and amino acid residue Tyr 225 of the human C1qC as shown in SEQ ID NO:3 or an amino acid residue of a human C1qC corresponding to Tyr 225 as shown in SEQ ID NO:3.
  • the anti-C1q antibody binds to amino acid residue Lys 219 of the human C1qA as shown in SEQ ID NO:1 or an amino acid residue of a human C1qA corresponding to Lys 219 as shown in SEQ ID NO:1, and amino acid residue Ser 185 of the human C1qC as shown in SEQ ID NO:3 or an amino acid residue of a human C1qC corresponding to Ser 185 as shown in SEQ ID NO:3.
  • the anti-C1q antibody binds to a C1q protein and binds to one or more amino acids of the C1q protein within amino acid residues selected from: (a) amino acid residues 218-240 of SEQ ID NO:3 (SEQ ID NO:10) or amino acid residues of a C1qC corresponding to amino acid residues 218-240 (WLAVNDYYDMVGI QGSDSVFSGF) of SEQ ID NO:3 (SEQ ID NO:10); (b) amino acid residues 225-240 of SEQ ID NO:3 (SEQ ID NO:11) or amino acid residues of a C1qC corresponding to amino acid residues 225-240 (YDMVGI QGSDSVFSGF) of SEQ ID NO:3 (SEQ ID NO:11); (c) amino acid residues 225-232 of SEQ ID NO:3 (SEQ ID NO:12) or amino acid residues of a C1qC corresponding to amino acid residues 225-232 (YDMVGIQ
  • 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 interaction between C1q and C1s and between C1q and C1r. In some embodiments, the anti-C1q antibody inhibits the interaction between C1q and another antibody, such as an autoantibody. In some embodiments, the anti-C1q antibody inhibits the respective interactions, at a stoichiometry of less than 2.5:1; 2.0:1; 1.5:1; or 1.0:1.
  • the C1q antibody inhibits an interaction, such as the C1q-C1s interaction, at approximately equimolar concentrations of C1q and the anti-C1q antibody.
  • the anti-C1q antibody binds to C1q with a stoichiometry of less than 20:1; less than 19.5:1; less than19:1; 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; less than 11:1; 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
  • the anti-C1q antibody binds C1q with a binding stoichiometry that ranges from 20:1 to 1.0:1 or less than1.0:1. In certain embodiments, the anti-C1q antibody binds C1q with a binding stoichiometry that ranges from 6:1 to 1.0:1 or less than1.0:1. In certain embodiments, the anti-C1q antibody binds C1q with a binding stoichiometry that ranges from 2.5:1 to 1.0:1 or less than1.0:1. 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.
  • 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 C1q A-chain. In other embodiments, the anti-C1q antibody binds to the C1q B-chain. In other embodiments, the anti-C1q antibody binds to the C1q C-chain. In some embodiments, the anti-C1q antibody binds to the C1q A-chain, the C1q B-chain and/or the C1q C-chain.
  • the anti-C1q antibody binds to the globular domain of the C1q A-chain, B-chain, and/or C-chain. In other embodiments, the anti-C1q antibody binds to the collagen-like domain of the C1q A-chain, the C1q B-chain, and/or the C1q C-chain.
  • 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% relative to a control wherein the antibodies of this disclosure are absent.
  • the interaction occurring in the presence of the antibody is reduced by an amount that ranges from at least 30% to at least 99% relative to a control wherein the antibodies of this disclosure are absent.
  • the antibodies of this disclosure inhibit 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 by an amount that ranges from at least 30% to at least 99%, relative to a control wherein the antibodies of this disclosure are absent.
  • Methods for measuring C4-cleavage are well known in the art.
  • the EC 50 values for antibodies of this disclosure with respect C4-cleavage may be less than 3 ⁇ g/ml; 2.5 ⁇ g/ml; 2.0 ⁇ g/ml; 1.5 ⁇ g/ml; 1.0 ⁇ g/ml; 0.5 ⁇ g/ml; 0.25 ⁇ g/ml; 0.1 ⁇ g/ml; 0.05 ⁇ g/ml.
  • the antibodies of this disclosure inhibit C4-cleavage at approximately equimolar concentrations of C1q and the respective anti-C1q antibody.
  • the antibodies of this disclosure inhibit autoantibody-dependent and complement-dependent 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 by an amount that ranges from at least 30% to at least 99%, relative to a control wherein the antibodies of this disclosure are absent.
  • CDC autoantibody-dependent and complement-dependent cytotoxicity
  • the EC 50 values for antibodies of this disclosure with respect to inhibition of autoantibody-dependent and complement-dependent cytotoxicity may be less than 3 ⁇ g/ml; 2.5 ⁇ g/ml; 2.0 ⁇ g/ml; 1.5 ⁇ g/ml; 1.0 ⁇ g/ml; 0.5 ⁇ g/ml; 0.25 ⁇ g/ml; 0.1 ⁇ g/ml; 0.05 ⁇ g/ml.
  • the antibodies of this 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 by an amount that ranges from at least 30% to at least 99%, relative to a control wherein the antibodies of this disclosure are absent.
  • CDCC complement-dependent cell-mediated cytotoxicity
  • the EC 50 values for antibodies of this disclosure with respect CDCC inhibition may be 1 less than 3 ⁇ g/ml; 2.5 ⁇ g/ml; 2.0 ⁇ g/ml; 1.5 g/ml; 1.0 ⁇ g/ml; 0.5 ⁇ g/ml;0.25 ⁇ g/ml; 0.1 ⁇ g/ml; 0.05 ⁇ g/ml.
  • the antibodies of this disclosure inhibit CDCC but not antibody-dependent cellular cytotoxicity (ADCC).
  • the antibodies of this disclosure inhibit C1F hemolysis (also referred to as CH50 hemolysis) 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 by an amount that ranges from at least 30% to at least 99%,relative to a control wherein the antibodies of this disclosure are absent or wherein control antibodies are used that do not bind to a complement factor or another antibody such as an autoantibody (see, e.g., Example 3).
  • Methods for measuring C1F hemolysis are well known in the art (see, e.g., Example 3).
  • the EC 50 values for antibodies of this disclosure with respect to C1F hemolysis may be less than 3 ⁇ g/ml; 2.5 ⁇ g/ml; 2.0 ⁇ g/ml; 1.5 ⁇ g/ml; 1.0 ⁇ g/ml; 0.5 ⁇ g/ml; 0.25 ⁇ g/ml; 0.1 ⁇ g/ml; 0.05 ⁇ g/ml.
  • the anti-C1q antibodies of this disclosure neutralize at least 50% of C1F hemolysis at a dose of less than 200 ng/ml, less than100 ng/ml, less than 50 ng/ml, or less than 20 ng/ml.
  • the antibodies of this disclosure neutralize C1F hemolysis at approximately equimolar concentrations of C1q and the anti-C1q antibody. In some embodiments, the anti-C1q antibodies of this disclosure neutralize hemolysis in a human C1F hemolysis assay. In some embodiments, the antibodies of this disclosure neutralize hemolysis in a human, mouse, and rat C1F hemolysis assay (see, e.g., Example 3).
  • the alternative pathway may amplify CDC initiated by C1q binding and subsequent C1s activation; in at least some of these embodiments, the antibodies of this disclosure inhibit the alternative pathway by 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 by an amount that ranges from at least 30% to at least 99%, relative to a control wherein the antibodies of this disclosure were absent.
  • the antibodies of this disclosure prevent synaptic loss in a cellular in vitro model or an in vivo model of synaptic loss, such as an in vivo mouse model.
  • In vivo mouse models may include Tg2576, a mouse amyloid precursor protein (APP) transgenic model of Alzheimer's disease, R6/2 NT-CAG150, a transgenic model for Huntington's disease, or SMA ⁇ 7, a mouse model for Spinal Muscular Atrophy, or DBA/2J, a genetic mouse model of glaucoma.
  • APP mouse amyloid precursor protein
  • any neurodegenerative disease model such as an Alzheimer's disease and/or Huntington's disease model, may be used that displays synapse loss.
  • In vitro lesion formation may be reduced by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, or by an amount that ranges from at least 30% to at least 95%, relative to a control experiment in which antibodies of this disclosure are absent.
  • the EC 50 values for antibodies of this disclosure with respect to the prevention of in vitro lesion formation may be less than 3 ⁇ g/ml; 2.5 ⁇ g/ml; 2.0 ⁇ g/ml; 1.5 ⁇ g/ml; 1.0 ⁇ g/ml; 0.5 ⁇ g/ml; 0.25 ⁇ g/ml; 0.1 ⁇ g/ml; 0.05 ⁇ g/ml.
  • In vivo synaptic loss may be reduced by at least 5%, at least 10%, at least 15%, at least 20%, at least 35%, at least 40%, or at least 50%, or by an amount that ranges from at least 5% to at least 50%, relative to a control experiment in which antibodies of this disclosure are absent.
  • the present disclosure provides anti-C1q antibodies.
  • the antibodies of this disclosure may have one or more of the following characteristics.
  • the antibodies of this disclosure may be polyclonal antibodies, monoclonal antibodies, humanized antibodies, human antibodies, antibody fragments, bispecific and polyspecific antibodies, multivalent antibodies, or heteroconjugate antibodies.
  • Antibody fragments of this disclosure may be functional fragments that bind the same epitope as any of the anti-C1q antibodies of this disclosure.
  • the antibody fragments of this disclosure specifically bind to and neutralize a biological activity of C1q.
  • the antibody fragments are miniaturized versions of the anti-C1q antibodies or antibody fragments of this disclosure that have the same epitope of the corresponding full-length antibody, but have much smaller molecule weight.
  • anti-C1q antibody fragments of this disclosure may have better brain penetration ability and a shorter half-life, which is advantageous for imaging and diagnostic utilities (see e.g., Lütje S et al., Bioconjug Chem. 2014 Feb 19; 25(2):335-41; Tavaré R et al., Proc Natl Acad Sci USA. 2014 Jan. 21; 111(3):1108-13; and Wiehr S et al., Prostate. 2014 May; 74(7):743-55). Accordingly, in some embodiments, anti-C1q antibody fragments of this disclosure have better brain penetration as compared to their corresponding full-length antibodies and/or have a shorter half-life as compared to their corresponding full-length antibodies.
  • anti-C1q antibodies of the present disclosure are bispecific antibodies recognizing a first antigen and a second antigen.
  • the first antigen is a C1q antigen.
  • the second antigen is an antigen facilitating transport across the blood-brain-barrier, including without limitation, transferrin receptor (TR), insulin receptor (HIR), insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor related proteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, CRM197, a llama single domain antibody, TMEM 30(A), a protein transduction domain, TAT, Syn-B, penetratin, a poly-arginine peptide, an angiopep peptide, and ANG1005.
  • the antibodies of this disclosure may further contain engineered effector functions, amino acid sequence modifications or other antibody modifications known in the art; e.g., the constant region of the anti-C1q antibodies described herein may be modified to impair complement activation.
  • the anti-C1q antibodies of this disclosure prevent Alzheimer's disease and/or Huntington's disease, or one or more symptoms of such neurodegenerative diseases.
  • prevention of Alzheimer's disease and/or Huntington's disease, or one or more symptoms of such neurodegenerative diseases by the anti-C1q antibodies of the present disclosure may be measured by inhibition of A ⁇ -induced C3 deposition and/or inhibit synapse loss in the hippocampus in an in vivo mouse model of Alzheimer's disease and/or Huntington's disease.
  • the anti-C1q antibodies of this disclosure inhibit A ⁇ -induced C3 deposition and/or inhibit synapse loss in the hippocampus in an in vivo mouse model of Alzheimer's disease and/or Huntington's disease.
  • Methods for measuring A ⁇ -induced C3 deposition and/or inhibit synapse loss in the hippocampus in vivo are well known in the art (see also Examples 5-9 for exemplary methods)
  • Additional anti-C1q antibodies e.g., antibodies that specifically bind to a C1q protein of the present disclosure, may be identified, screened, and/or characterized for their physical/chemical properties and/or biological activities by various assays known in the art.
  • Anti-C1q antibodies of the present disclosure can encompass polyclonal antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies, human antibodies, antibody fragments (e.g., Fab, Fab′-SH, Fv, scFv, and F(ab′) 2 fragments), bispecific and polyspecific antibodies, multivalent antibodies, heteroconjugate antibodies, library derived antibodies, antibodies having modified effector functions, fusion proteins containing an antibody portion, and any other modified configuration of the immunoglobulin molecule that includes an antigen recognition site, such as an epitope having amino acid residues of a C1q protein of the present disclosure, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies.
  • the anti-C1q antibodies may be human, murine, rat, or of any other origin (including chimeric or humanized antibodies).
  • Polyclonal antibodies such as polyclonal anti-C1q antibodies, are generally raised in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen (e.g., a purified or recombinant C1q protein of the present disclosure) to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin (KLH), serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor, using a bifunctional or derivatizing agent, e.g., maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl 2 , or R 1 N ⁇ C ⁇ NR, where R and R 1 are independently lower al
  • adjuvants examples include Freund's complete adjuvant and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).
  • the immunization protocol may be selected by one skilled in the art without undue experimentation.
  • the animals are immunized against the desired antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 ⁇ g (for rabbits) or 5 ⁇ g (for mice) of the protein or conjugate with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites.
  • the animals are boosted with 1 ⁇ 5 to 1/10 the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites.
  • the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus.
  • Conjugates also can be made in recombinant-cell culture as protein fusions. Also, aggregating agents such as alum are suitable to enhance the immune response.
  • Monoclonal antibodies such as monoclonal anti-C1q antibodies, are obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and/or post-translational modifications (e.g., isomerizations, amidations) that may be present in minor amounts.
  • the modifier “monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies.
  • the monoclonal anti-C1q antibodies may be made using the hybridoma method first described by Köhler et al., Nature, 256:495 (1975), or may be made by recombinant DNA methods (U.S. Pat. No. 4,816,567).
  • a mouse or other appropriate host animal such as a hamster
  • lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization (e.g., a purified or recombinant C1q protein of the present disclosure).
  • lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice , pp. 59-103 (Academic Press, 1986)).
  • the immunizing agent will typically include the antigenic protein (e.g., a purified or recombinant C1q protein of the present disclosure) or a fusion variant thereof.
  • the antigenic protein e.g., a purified or recombinant C1q protein of the present disclosure
  • a fusion variant thereof e.g., peripheral blood lymphocytes (“PBLs”) are used if cells of human origin are desired, while spleen or lymph node cells are used if non-human mammalian sources are desired.
  • PBLs peripheral blood lymphocytes
  • the lymphoctyes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell. Goding, Monoclonal Antibodies: Principles and Practice , Academic Press (1986), pp. 59-103.
  • Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine or human origin. Usually, rat or mouse myeloma cell lines are employed.
  • the hybridoma cells thus prepared are seeded and grown in a suitable culture medium that may contain one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
  • the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which are substances that prevent the growth of HGPRT-deficient-cells.
  • HGPRT hypoxanthine guanine phosphoribosyl transferase
  • immortalized myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium.
  • murine myeloma lines such as those derived from MOPC-21 and MPC-11 mouse tumors (available from the Salk Institute Cell Distribution Center, San Diego, Calif. USA), as well as SP-2 cells and derivatives thereof (e.g., X63-Ag8-653) (available from the American Type Culture Collection, Manassas, Va. USA).
  • Human myeloma and mouse-human heteromyeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor, J. Immunol. , 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications , pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
  • Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen (e.g., a C1q protein of the present disclosure).
  • the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA).
  • the culture medium in which the hybridoma cells are cultured can be assayed for the presence of monoclonal antibodies directed against the desired antigen (e.g., a C1q protein of the present disclosure).
  • the binding affinity and specificity of the monoclonal antibody can be determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked assay (ELISA).
  • RIA radioimmunoassay
  • ELISA enzyme-linked assay
  • binding affinity may be determined by the Scatchard analysis of Munson et al., Anal. Biochem., 107:220 (1980).
  • the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, supra). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium.
  • the hybridoma cells may be grown in vivo as tumors in a mammal.
  • the monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose chromatography, hydroxylapatite chromatography, gel electrophoresis, dialysis, affinity chromatography, and other methods as described above.
  • Anti-C1q monoclonal antibodies may also be made by recombinant DNA methods, such as those disclosed in U.S. Pat. No. 4,816,567, and as described above.
  • DNA encoding the monoclonal antibodies is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that specifically bind to genes encoding the heavy and light chains of murine antibodies).
  • the hybridoma cells serve as a source of such DNA. Once isolated, the DNA may be placed into expression vectors, which are then transfected into host-cells such as E.
  • anti-C1q antibodies can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al., Nature, 348:552-554 (1990). Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991) described the isolation of murine and human antibodies, respectively, from phage libraries.
  • DNA encoding antibodies or fragments thereof may also be modified, for example, by substituting the coding sequence for human heavy- and light-chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, et al., Proc. Natl Acad. Sci. USA, 81:6851 (1984)), or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
  • non-immunoglobulin polypeptides are substituted for the constant domains of an antibody, or they are substituted for the variable domains of one antigen-combining site of an antibody to create a chimeric bivalent antibody comprising one antigen-combining site having specificity for an antigen and another antigen-combining site having specificity for a different antigen.
  • the monoclonal antibodies described herein may by monovalent, the preparation of which is well known in the art.
  • one method involves recombinant expression of immunoglobulin light chain and a modified heavy chain.
  • the heavy chain is truncated generally at any point in the Fc region so as to prevent heavy chain crosslinking.
  • the relevant cysteine residues may be substituted with another amino acid residue or are deleted so as to prevent crosslinking.
  • In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce fragments thereof, particularly Fab fragments, can be accomplished using routine techniques known in the art.
  • Chimeric or hybrid anti-C1q antibodies also may be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents.
  • immunotoxins may be constructed using a disulfide-exchange reaction or by forming a thioether bond.
  • suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate.
  • Anti-C1q antibodies of the present disclosure or antibody fragments thereof may further include humanized or human antibodies.
  • Humanized forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fab, Fab′-SH, Fv, scFv, F(ab′) 2 or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin.
  • Humanized antibodies include human immunoglobulins (recipient antibody) in which residues from a complementarity determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity.
  • CDR complementarity determining region
  • donor antibody non-human species
  • Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence.
  • the humanized antibody optimally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. Jones et al., Nature 321: 522-525 (1986); Riechmann et al., Nature 332: 323-329 (1988) and Presta, Curr. Opin. Struct. Biol. 2: 593-596 (1992).
  • the anti-C1q antibody is a chimeric antibody comprising the heavy and light chain variable domains of any of the anti-C1q antibody described herein (e.g., antibody M1 and 4A4B 11) and constant regions from a human immunoglobulin.
  • a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Humanization can be essentially performed following the method of Winter and co-workers, Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988), or through substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody.
  • humanized antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species.
  • humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
  • variable domains both light and heavy
  • sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences.
  • the human sequence which is closest to that of the rodent is then accepted as the human framework (FR) for the humanized antibody.
  • FR human framework
  • Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. The same framework may be used for several different humanized antibodies. Carter et al., Proc. Nat'l Acad. Sci. USA 89:4285 (1992); Presta et al., J. Immunol. 151:2623 (1993).
  • humanized antibodies are prepared by a process of analyzing the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences.
  • Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art.
  • Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen.
  • FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen or antigens (e.g., C1q proteins of the present disclosure), is achieved.
  • the CDR residues are directly and most substantially involved in influencing antigen binding.
  • the humanized anti-C1q antibody may be an antibody fragment, such as an Fab, which is optionally conjugated with one or more cytotoxic agent(s) in order to generate an immunoconjugate.
  • the humanized anti-C1q antibody may be an intact antibody, such as an intact IgG1 antibody.
  • human anti-C1q antibodies can be generated.
  • transgenic animals e.g., mice
  • transgenic animals e.g., mice
  • the homozygous deletion of the antibody heavy-chain joining region (J H ) gene in chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production.
  • Transfer of the human germ-line immunoglobulin gene array in such germ-line mutant mice will result in the production of human antibodies upon antigen challenge. See, e.g., Jakobovits et al., Proc. Nat'l Acad. Sci.
  • phage display technology can be used to produce human anti-C1q antibodies and antibody fragments in vitro, from immunoglobulin variable (V) domain gene repertoires from unimmunized donors. McCafferty et al., Nature 348:552-553 (1990); Hoogenboom and Winter, J. Mol. Biol. 227: 381 (1991).
  • antibody V domain genes are cloned in-frame into either a major or minor coat protein gene of a filamentous bacteriophage, such as M13 or fd, and displayed as functional antibody fragments on the surface of the phage particle.
  • the filamentous particle contains a single-stranded DNA copy of the phage genome, selections based on the functional properties of the antibody also result in selection of the gene encoding the antibody exhibiting those properties.
  • the phage mimics some of the properties of the B-cell.
  • Phage display can be performed in a variety of formats, reviewed in, e.g., Johnson, Kevin S. and Chiswell, David J., Curr. Opin Struct. Biol. 3:564-571 (1993).
  • V-gene segments can be used for phage display.
  • yeast display technology can be used to produce human anti-C1q antibodies and antibody fragments in vitro (e.g., WO 2009/036379; WO 2010/105256; WO 2012/009568; US 2009/0181855; US 2010/0056386; and Feldhaus and Siegel (2004) J. Immunological Methods 290:69-80).
  • ribosome display technology can be used to produce human anti-C1q antibodies and antibody fragments in vitro (e.g., Roberts and Szostak (1997) Proc Natl Acad Sci 94:12297-12302; Schaffitzel et al. (1999) J. Immunolical Methods 231:119-135; Lipovsek and Pl ückthun (2004) J. Immunological Methods 290:51-67).
  • human anti-C1q monoclonal antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly and antibody repertoire.
  • human anti-C1q antibodies may also be generated in vitro by activated B-cells (see U.S. Pat. Nos. 5,567,610 and 5,229,275).
  • a anti-C1q antibody fragments can also be isolated from the antibody phage libraries as discussed above.
  • Fab′-SH fragments can be directly recovered from E. coli and chemically coupled to form F(ab′) 2 fragments (Carter et al., Bio/Technology 10:163-167 (1992)).
  • F(ab′) 2 fragments can be isolated directly from recombinant host-cell culture. Production of Fab and F(ab′) 2 antibody fragments with increased in vivo half-lives are described in U.S. Pat. No. 5,869,046.
  • the antibody of choice is a single chain Fv fragment (scFv). See WO 93/16185; U.S. Pat.
  • the anti-C1q, anti-C1r , or anti-C1q antibody fragment may also be a “linear antibody,” e.g., as described in U.S. Pat. No. 5,641,870. Such linear antibody fragments may be monospecific or bispecific.
  • Bispecific antibodies are antibodies that have binding specificities for at least two different epitopes, including those on the same or another protein (e.g., one or more C1q proteins of the present disclosure). Alternatively, one part of a BsAb can be armed to bind to the target C1q antigen, and another can be combined with an arm that binds to a second protein. Such antibodies can be derived from full length antibodies or antibody fragments (e.g., F(ab′) 2 bispecific antibodies).
  • bispecific antibodies are known in the art. Traditional production of full length bispecific antibodies is based on the coexpression of two immunoglobulin heavy-chain/light chain pairs, where the two chains have different specificities. Millstein et al., Nature, 305:537-539 (1983). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of 10 different antibody molecules, of which only one has the correct bispecific structure. Purification of the correct molecule, which is usually done by affinity chromatography steps, is rather cumbersome, and the product yields are low. Similar procedures are disclosed in WO 93/08829 and in Traunecker et al., EMBO J., 10:3655-3659 (1991).
  • antibody variable domains with the desired binding specificities are fused to immunoglobulin constant domain sequences.
  • the fusion may be with an immunoglobulin heavy chain constant domain, comprising at least part of the hinge, C H 2, and C H 3 regions.
  • the first heavy-chain constant region (C H 1) containing the site necessary for light chain binding is present in at least one of the fusions.
  • the bispecific antibodies are composed of a hybrid immunoglobulin heavy chain with a first binding specificity in one arm, and a hybrid immunoglobulin heavy chain-light chain pair (providing a second binding specificity) in the other arm. It was found that this asymmetric structure facilitates the separation of the desired bispecific compound from unwanted immunoglobulin chain combinations, as the presence of an immunoglobulin light chain in only half of the bispecific molecules provides for an easy way of separation. This approach is disclosed in WO 94/04690. For further details of generating bispecific antibodies, see, for example, Suresh et al., Methods in Enzymology 121: 210 (1986).
  • the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant-cell culture.
  • the interface may comprise at least a part of the C H 3 region of an antibody constant domain.
  • one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g., tyrosine or tryptophan).
  • Compensatory “cavities” of identical or similar size to the large side chains(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g., alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.
  • bispecific antibodies can be prepared using chemical linkage.
  • Brennan et al., Science 229:81 (1985) describe a procedure wherein intact antibodies are proteolytically cleaved to generate F(ab′) 2 fragments. These fragments are reduced in the presence of the dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation.
  • the Fab′ fragments generated are then converted to thionitrobenzoate (TNB) derivatives.
  • TAB thionitrobenzoate
  • One of the Fab′-TNB derivatives is then reconverted to the Fab′-TNB derivative to form the bispecific antibody.
  • the bispecific antibodies produced can be used as agents for the selective immobilization of enzymes.
  • Fab′ fragments may be directly recovered from E. coli and chemically coupled to form bispecific antibodies.
  • Shalaby et al., J. Exp. Med. 175: 217-225 (1992) describes the production of fully humanized bispecific antibody F(ab′) 2 molecules.
  • Each Fab′ fragment was separately secreted from E. coli and subjected to directed chemical coupling in vitro to form the bispecific antibody.
  • the bispecific antibody thus formed was able to bind to cells overexpressing the ErbB2 receptor and normal human T-cells, as well as trigger the lytic activity of human cytotoxic lymphocytes against human breast tumor targets.
  • bivalent heterodimers have been produced using leucine zippers.
  • the leucine zipper peptides from the Fos and Jun proteins were linked to the Fab′ portions of two different antibodies by gene fusion.
  • the antibody homodimers were reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers.
  • the “diabody” technology described by Hollinger et al., Proc. Nat'l Acad. Sci.
  • the fragments comprise a heavy-chain variable domain (V H ) connected to a light-chain variable domain (V L ) by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the V H and V L , domains of one fragment are forced to pair with the complementary V L , and V H domains of another fragment, thereby forming two antigen-binding sites.
  • V H and V L , domains of one fragment are forced to pair with the complementary V L , and V H domains of another fragment, thereby forming two antigen-binding sites.
  • sFv single-chain Fv
  • Antibodies with more than two valencies are also contemplated.
  • trispecific antibodies can be prepared. Tutt et al., J. Immunol. 147:60 (1991).
  • Exemplary bispecific antibodies may bind to two different antigens.
  • a bispecific antibody binds to a first antigen, C1q, and a second antigen facilitating transport across the blood-brain barrier.
  • C1q a first antigen
  • second antigen facilitating transport across the blood-brain barrier.
  • Numerous antigens are known in the art that facilitate transport across the blood-brain barrier (see, e.g., Gabathuler R., Approaches to transport therapeutic drugs across the blood-brain barrier to treat brain diseases, Neurobiol. Dis. 37 (2010) 48-57).
  • Such second antigens include, without limitation, transferrin receptor (TR), insulin receptor (HIR), Insulin-like growth factor receptor (IGFR), low-density lipoprotein receptor related proteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, including CRM197 (a non-toxic mutant of diphtheria toxin), llama single domain antibodies such as TMEM 30(A) (Flippase), protein transduction domains such as TAT, Syn-B, or penetratin, poly-arginine or generally positively charged peptides, and Angiopep peptides such as ANG1005 (see, e.g., Gabathuler, 2010).
  • TR transferrin receptor
  • HIR insulin receptor
  • IGFR Insulin-like growth factor receptor
  • LPR-1 and 2 low-density lipoprotein receptor related proteins 1 and 2
  • diphtheria toxin receptor including CRM197 (a non-toxic mutant of diphtheria toxin)
  • llama single domain antibodies such
  • a multivalent antibody may be internalized (and/or catabolized) faster than a bivalent antibody by a cell expressing an antigen to which the antibodies bind.
  • the anti-C1q antibodies of the present disclosure or antibody fragments thereof can be multivalent antibodies (which are other than of the IgM class) with three or more antigen binding sites (e.g., tetravalent antibodies), which can be readily produced by recombinant expression of nucleic acid encoding the polypeptide chains of the antibody.
  • the multivalent antibody can comprise a dimerization domain and three or more antigen binding sites.
  • the dimerization domain comprises an Fc region or a hinge region.
  • the antibody will comprise an Fc region and three or more antigen binding sites amino-terminal to the Fc region.
  • the multivalent antibody herein contains three to about eight, and in some embodiments four, antigen binding sites.
  • the multivalent antibody contains at least one polypeptide chain (and in some embodiments two polypeptide chains), wherein the polypeptide chain or chains comprise two or more variable domains.
  • the polypeptide chain or chains may comprise VD1-(X1)n-VD2-(X2)n-Fc, wherein VD1 is a first variable domain, VD2 is a second variable domain, Fc is one polypeptide chain of an Fc region, X1 and X2 represent an amino acid or polypeptide, and n is 0 or 1.
  • the polypeptide chain or chains may comprise V H -C H 1-flexible linker-V H -C H 1-Fc region chain; or V H -C H 1-V H -C H 1-Fc region chain.
  • the multivalent antibody herein may further comprise at least two (and in some embodiments four) light chain variable domain polypeptides.
  • the multivalent antibody herein may, for instance, comprise from about two to about eight light chain variable domain polypeptides.
  • the light chain variable domain polypeptides contemplated here comprise a light chain variable domain and, optionally, further comprise a CL domain.
  • Heteroconjugate antibodies are also within the scope of the present disclosure.
  • Heteroconjugate antibodies are composed of two covalently joined antibodies (e.g., anti-C1q antibodies of the present disclosure or antibody fragments thereof).
  • one of the antibodies in the heteroconjugate can be coupled to avidin, the other to biotin.
  • Such antibodies have, for example, been proposed to target immune system cells to unwanted cells, U.S. Pat. No. 4,676,980, and have been used to treat HIV infection.
  • International Publication Nos. WO 91/00360, WO 92/200373 and EP 0308936 It is contemplated that the antibodies may be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents.
  • immunotoxins may be constructed using a disulfide exchange reaction or by forming a thioether bond.
  • suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in U.S. Pat. No. 4,676,980.
  • Heteroconjugate antibodies may be made using any convenient cross-linking methods. Suitable cross-linking agents are well known in the art, and are disclosed in U.S. Pat. No. 4,676,980, along with a number of cross-linking techniques.
  • an anti-C1q antibody of the present disclosure may also be desirable to modify effector function and/or to increase serum half-life of the antibody.
  • the Fc receptor binding site on the constant region may be modified or mutated to remove or reduce binding affinity to certain Fc receptors, such as Fc ⁇ RI, Fc ⁇ RII, and/or Fc ⁇ RIII.
  • the effector function is impaired by removing N-glycosylation of the Fc region (e.g., in the CH 2 domain of IgG) of the antibody.
  • the effector function is impaired by modifying regions such as 233-236, 297, and/or 327-331 of human IgG as described in PCT WO 99/58572 and Armour et al., Molecular Immunology 40: 585-593 (2003); Reddy et al., J. Immunology 164:1925-1933 (2000).
  • the constant region of the anti-complement antibodies described herein may also be modified to impair complement activation.
  • complement activation of IgG antibodies following binding of the C1 component of complement may be reduced by mutating amino acid residues in the constant region in a C1 binding motif (e.g., C1q binding motif).
  • C1 binding motif e.g., C1q binding motif
  • C1s binding motif E318, K320, and K322 identified for murine IgG2b is believed to be common for other antibody isotypes (Duncan et al. (1988) Nature 322:738-740), C1q binding activity for IgG2b can be abolished by replacing any one of the three specified residues with a residue having an inappropriate functionality on its side chain. It is not necessary to replace the ionic residues only with Ala to abolish C1q binding.
  • non-ionic residues such as Gly, Ile, Leu, or Val
  • aromatic non-polar residues such as Phe, Tyr, Trp and Pro
  • polar non-ionic residues such as Ser, Thr, Cys, and Met in place of residues 320 and 322, but not 318, in order to abolish C1s binding activity.
  • Deglycosylation can be performed using glycosidase enzymes for example Endoglycosidase S (EndoS), a 108 kDa enzyme encoded by the gene endoS of Streptococcus pyogenes that selectively digests asparagine-linked glycans on the heavy chain of all IgG subclasses, without action on other immunoglobulin classes or other glycoproteins (Collin et al. (2001) EMBO J 2001,20:3046-3055).
  • EndoS Endoglycosidase S
  • Streptococcus pyogenes a 108 kDa enzyme encoded by the gene endoS of Streptococcus pyogenes that selectively digests asparagine-linked glycans on the heavy chain of all IgG subclasses, without action on other immunoglobulin classes or other glycoproteins
  • a salvage receptor binding epitope refers to an epitope of the Fc region of an IgG molecule (e.g., IgG 1 , IgG 2 , IgG 3 , or IgG 4 ) that is responsible for increasing the in vivo serum half-life of the IgG molecule.
  • Amino acid sequence modifications of anti-C1q antibodies of the present disclosure, or antibody fragments thereof, are also contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibodies or antibody fragments.
  • Amino acid sequence variants of the antibodies or antibody fragments are prepared by introducing appropriate nucleotide changes into the nucleic acid encoding the antibodies or antibody fragments, or by peptide synthesis. Such modifications include, for example, deletions from, and/or insertions into and/or substitutions of, residues within the amino acid sequences of the antibody.
  • Any combination of deletion, insertion, and substitution is made to arrive at the final construct, provided that the final construct possesses the desired characteristics (i.e., the ability to bind or physically interact with a C1q protein of the present disclosure).
  • the amino acid changes also may alter post-translational processes of the antibody, such as changing the number or position of glycosylation sites.
  • a useful method for identification of certain residues or regions of the anti-C1q antibody that are preferred locations for mutagenesis is called “alanine scanning mutagenesis” as described by Cunningham and Wells in Science, 244:1081-1085 (1989).
  • a residue or group of target residues are identified (e.g., charged residues such as arg, asp, his, lys, and glu) and replaced by a neutral or negatively charged amino acid (most preferably alanine or polyalanine) to affect the interaction of the amino acids with the target antigen.
  • Those amino acid locations demonstrating functional sensitivity to the substitutions then are refined by introducing further or other variants at, or for, the sites of substitution.
  • the site for introducing an amino acid sequence variation is predetermined, the nature of the mutation per se need not be predetermined. For example, to analyze the performance of a mutation at a given site, alanine scanning or random mutagenesis is conducted at the target codon or region and the expressed antibody variants are screened for the desired activity.
  • Amino acid sequence insertions include amino-(“N”) and/or carboxy-(“C”) terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues.
  • terminal insertions include an antibody with an N-terminal methionyl residue or the antibody fused to a cytotoxic polypeptide.
  • Other insertional variants of the antibody molecule include the fusion to the N- or C-terminus of the antibody to an enzyme or a polypeptide which increases the serum half-life of the antibody.
  • variants are an amino acid substitution variant. These variants have at least one amino acid residue in the antibody molecule replaced by a different residue.
  • the sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but FR alterations are also contemplated. Conservative substitutions are shown in the Table A below under the heading of “preferred substitutions”. If such substitutions result in a change in biological activity, then more substantial changes, denominated “exemplary substitutions” in Table A, or as further described below in reference to amino acid classes, may be introduced and the products screened.
  • Substantial modifications in the biological properties of the antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain.
  • Naturally occurring residues are divided into groups based on common side-chain properties:
  • hydrophobic norleucine, met, ala, val, leu, ile
  • Non-conservative substitutions entail exchanging a member of one of these classes for another class.
  • cysteine residue not involved in maintaining the proper conformation of the antibody also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant crosslinking.
  • cysteine bond(s) may be added to the antibody to improve its stability (particularly where the antibody is an antibody fragment, such as an Fv fragment).
  • the substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g. a humanized or human anti-C1q antibody).
  • a parent antibody e.g. a humanized or human anti-C1q antibody.
  • the resulting variant(s) selected for further development will have improved biological properties relative to the parent antibody from which they are generated.
  • a convenient way for generating such substitutional variants involves affinity maturation using phage display. Briefly, several hypervariable region sites (e.g., 6-7 sites) are mutated to generate all possible amino substitutions at each site.
  • the antibody variants thus generated are displayed in a monovalent fashion from filamentous phage particles as fusions to the gene III product of M13 packaged within each particle.
  • the phage-displayed variants are then screened for their biological activity (e.g., binding affinity) as herein disclosed.
  • alanine scanning mutagenesis can be performed to identify hypervariable region residues contributing significantly to antigen binding.
  • the panel of variants is subjected to screening as described herein and antibodies with superior properties in one or more relevant assays may be selected for further development.
  • Another type of amino acid variant of the antibody alters the original glycosylation pattern of the antibody. By altering is meant deleting one or more carbohydrate moieties found in the antibody, and/or adding one or more glycosylation sites that are not present in the antibody.
  • N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue.
  • the tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain.
  • X is any amino acid except proline
  • 0-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.
  • glycosylation sites to the antibody is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites).
  • the alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).
  • Nucleic acid molecules encoding amino acid sequence variants of the anti-IgE antibody are prepared by a variety of methods known in the art. These methods include, but are not limited to, isolation from a natural source (in the case of naturally occurring amino acid sequence variants) or preparation by oligonucleotide-mediated (or site-directed) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared variant or a non-variant version of the antibodies (e.g., anti-C1q antibody of the present disclosure) or antibody fragments.
  • Anti-C1q antibodies of the present disclosure, or antibody fragments thereof, can be further modified to contain additional non-proteinaceous moieties that are known in the art and readily available.
  • the moieties suitable for derivatization of the antibody are water-soluble polymers.
  • Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene/maleic anhydride copolymer, polyaminoacids (either homopolymers or random copolymers), and dextran or poly(n-vinyl pyrrolidone)polyethylene glycol, polypropylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof.
  • PEG polyethylene glycol
  • copolymers of ethylene glycol/propylene glycol carboxymethylcellulose
  • dextran polyvinyl alcohol
  • Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water.
  • the polymer may be of any molecular weight, and may be branched or unbranched.
  • the number of polymers attached to the antibody may vary, and if more than one polymer is attached, they can be the same or different molecules. In general, the number and/or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular properties or functions of the antibody to be improved, whether the antibody derivative will be used in a therapy under defined conditions, etc.
  • Such techniques and other suitable formulations are disclosed in Remington: The Science and Practice of Pharmacy, 20th Ed., Alfonso Gennaro, Ed., Philadelphia College of Pharmacy and Science (2000).
  • Anti-C1q antibodies of the present disclosure may be produced using recombinant methods and compositions, e.g., as described in U.S. Pat. No. 4,816,567.
  • isolated nucleic acids having a nucleotide sequence encoding any of the anti-C1q antibodies of the present disclosure are provided.
  • Such nucleic acids may encode an amino acid sequence containing the VL and/or an amino acid sequence containing the VH of the anti-C1q antibody (e.g., the light and/or heavy chains of the antibody).
  • one or more vectors (e.g., expression vectors) containing such nucleic acids are provided.
  • a host cell containing such nucleic acid is also provided.
  • the host cell contains (e.g., has been transduced with): (1) a vector containing a nucleic acid that encodes an amino acid sequence containing the VL of the antibody and an amino acid sequence containing the VH of the antibody, or (2) a first vector containing a nucleic acid that encodes an amino acid sequence containing the VL of the antibody and a second vector containing a nucleic acid that encodes an amino acid sequence containing the VH of the antibody.
  • the host cell is eukaryotic, e.g., a Chinese Hamster Ovary (CHO) cell or lymphoid cell (e.g., Y0, NS0, Sp20 cell).
  • the method includes culturing a host cell of the present disclosure containing a nucleic acid encoding the anti-C1q antibody, under conditions suitable for expression of the antibody.
  • the antibody is subsequently recovered from the host cell (or host cell culture medium). See also Example 1.
  • a nucleic acid encoding the anti-C1q antibody is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell.
  • Such nucleic acid may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the antibody).
  • Suitable vectors containing a nucleic acid sequence encoding any of the anti-C1q antibodies of the present disclosure, or fragments thereof polypeptides (including antibodies) described herein include, without limitation, cloning vectors and expression vectors.
  • Suitable cloning vectors can be constructed according to standard techniques, or may be selected from a large number of cloning vectors available in the art. While the cloning vector selected may vary according to the host cell intended to be used, useful cloning vectors generally have the ability to self-replicate, may possess a single target for a particular restriction endonuclease, and/or may carry genes for a marker that can be used in selecting clones containing the vector.
  • Suitable examples include plasmids and bacterial viruses, e.g., pUC18, pUC19, Bluescript (e.g., pBS SK+) and its derivatives, mp18, mp19, pBR322, pMB9, ColE1, pCR1, RP4, phage DNAs, and shuttle vectors such as pSA3 and pAT28.
  • Bluescript e.g., pBS SK+
  • shuttle vectors such as pSA3 and pAT28.
  • Expression vectors generally are replicable polynucleotide constructs that contain a nucleic acid of the present disclosure.
  • the expression vector may replicable in the host cells either as episomes or as an integral part of the chromosomal DNA.
  • Suitable expression vectors include but are not limited to plasmids, viral vectors, including adenoviruses, adeno-associated viruses, retroviruses, cosmids, and expression vector(s) disclosed in PCT Publication No. WO 87/04462.
  • Vector components may generally include, but are not limited to, one or more of the following: a signal sequence; an origin of replication; one or more marker genes; suitable transcriptional controlling elements (such as promoters, enhancers and terminator).
  • suitable transcriptional controlling elements such as promoters, enhancers and terminator
  • the vectors containing the nucleic acids of interest can be introduced into the host cell by any of a number of appropriate means, including electroporation, transfection employing calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, or other substances; microprojectile bombardment; lipofection; and infection (e.g., where the vector is an infectious agent such as vaccinia virus).
  • electroporation employing calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, or other substances
  • microprojectile bombardment e.g., where the vector is an infectious agent such as vaccinia virus.
  • infection e.g., where the vector is an infectious agent such as vaccinia virus.
  • the vector contains a nucleic acid containing one or more amino acid sequences encoding an anti-C1q antibody of the present disclosure.
  • Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells.
  • anti-C1q antibodies of the present disclosure may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed.
  • For expression of antibody fragments and polypeptides in bacteria e.g., U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523; and Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J., 2003), pp. 245-254, describing expression of antibody fragments in E. coli .).
  • the antibody may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.
  • eukaryotic microorganisms such as filamentous fungi or yeast
  • suitable cloning or expression hosts for antibody-encoding vectors including fungi and yeast strains whose glycosylation pathways have been “humanized,” resulting in the production of an antibody with a partially or fully human glycosylation pattern (e.g., Gerngross, Nat. Biotech. 22:1409-1414 (2004); and Li et al., Nat. Biotech. 24:210-215 (2006)).
  • Suitable host cells for the expression of glycosylated antibody can also be derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frupperda cells. Plant cell cultures can also be utilized as hosts (e.g., U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429, describing PLANTIBODIESTM technology for producing antibodies in transgenic plants.).
  • Vertebrate cells may also be used as hosts.
  • mammalian cell lines that are adapted to grow in suspension may be useful.
  • Other examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod.
  • monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells.
  • Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al., Proc. Natl. Acad. Sci.
  • Anti-C1q antibodies of the present disclosure can be incorporated into a variety of formulations for therapeutic use (e.g., by administration) or in the manufacture of a medicament (e.g., for treating or preventing a neurodegenerative disease, such as Alzheimer's disease or Huntington's disease) by combining the antibodies with appropriate pharmaceutically acceptable carriers or diluents, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms. Examples of such formulations include, without limitation, tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols.
  • compositions can include, depending on the formulation desired, pharmaceutically-acceptable, non-toxic carriers of diluents, which are vehicles commonly used to formulate pharmaceutical compositions for animal or human administration.
  • diluents are vehicles commonly used to formulate pharmaceutical compositions for animal or human administration.
  • the diluent is selected so as not to affect the biological activity of the combination.
  • examples of such diluents include, without limitation, distilled water, buffered water, physiological saline, PBS, Ringer's solution, dextrose solution, and Hank's solution.
  • a pharmaceutical composition or formulation of the present disclosure can further include other carriers, adjuvants, or non-toxic, nontherapeutic, nonimmunogenic stabilizers, excipients and the like.
  • the compositions can also include additional substances to approximate physiological conditions, such as pH adjusting and buffering agents, toxicity adjusting agents, wetting agents and detergents.
  • a pharmaceutical composition of the present disclosure can also include any of a variety of stabilizing agents, such as an antioxidant for example.
  • the pharmaceutical composition includes a polypeptide
  • the polypeptide can be complexed with various well-known compounds that enhance the in vivo stability of the polypeptide, or otherwise enhance its pharmacological properties (e.g., increase the half-life of the polypeptide, reduce its toxicity, and enhance solubility or uptake).
  • modifications or complexing agents include, without limitation, sulfate, gluconate, citrate and phosphate.
  • the polypeptides of a composition can also be complexed with molecules that enhance their in vivo attributes. Such molecules include, without limitation, carbohydrates, polyamines, amino acids, other peptides, ions (e.g., sodium, potassium, calcium, magnesium, manganese), and lipids.
  • the active ingredient can be administered in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions.
  • the active component(s) can be encapsulated in gelatin capsules together with inactive ingredients and powdered carriers, such as glucose, lactose, sucrose, mannitol, starch, cellulose or cellulose derivatives, magnesium stearate, stearic acid, sodium saccharin, talcum, magnesium carbonate.
  • inactive ingredients and powdered carriers such as glucose, lactose, sucrose, mannitol, starch, cellulose or cellulose derivatives, magnesium stearate, stearic acid, sodium saccharin, talcum, magnesium carbonate.
  • additional inactive ingredients that may be added to provide desirable color, taste, stability, buffering capacity, dispersion or other known desirable features are red iron oxide, silica gel, sodium lauryl sulfate, titanium dioxide, and edible white ink.
  • Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance.
  • Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives.
  • compositions intended for in vivo use are usually sterile. To the extent that a given compound must be synthesized prior to use, the resulting product is typically substantially free of any potentially toxic agents, particularly any endotoxins, which may be present during the synthesis or purification process.
  • compositions for parental administration are also sterile, substantially isotonic and made under GMP conditions.
  • Formulations may be optimized for retention and stabilization in the brain or central nervous system.
  • Stabilization techniques include cross-linking, multimerizing, or linking to groups such as polyethylene glycol, polyacrylamide, neutral protein carriers, etc. in order to achieve an increase in molecular weight.
  • Implants may be particles, sheets, patches, plaques, fibers, microcapsules and the like and may be of any size or shape compatible with the selected site of insertion.
  • the implants may be monolithic, i.e. having the active agent homogenously distributed through the polymeric matrix, or encapsulated, where a reservoir of active agent is encapsulated by the polymeric matrix.
  • the selection of the polymeric composition to be employed will vary with the site of administration, the desired period of treatment, patient tolerance, the nature of the disease to be treated and the like. Characteristics of the polymers will include biodegradability at the site of implantation, compatibility with the agent of interest, ease of encapsulation, a half-life in the physiological environment.
  • Biodegradable polymeric compositions which may be employed may be organic esters or ethers, which when degraded result in physiologically acceptable degradation products, including the monomers. Anhydrides, amides, orthoesters or the like, by themselves or in combination with other monomers, may find use.
  • the polymers will be condensation polymers.
  • the polymers may be cross-linked or non-cross-linked.
  • polymers of hydroxyaliphatic carboxylic acids either homo- or copolymers, and polysaccharides. Included among the polyesters of interest are polymers of D-lactic acid, L-lactic acid, racemic lactic acid, glycolic acid, polycaprolactone, and combinations thereof.
  • a slowly biodegrading polymer is achieved, while degradation is substantially enhanced with the racemate.
  • Copolymers of glycolic and lactic acid are of particular interest, where the rate of biodegradation is controlled by the ratio of glycolic to lactic acid.
  • the most rapidly degraded copolymer has roughly equal amounts of glycolic and lactic acid, where either homopolymer is more resistant to degradation.
  • the ratio of glycolic acid to lactic acid will also affect the brittleness of in the implant, where a more flexible implant is desirable for larger geometries.
  • polysaccharides of interest are calcium alginate, and functionalized celluloses, particularly carboxymethylcellulose esters characterized by being water insoluble, a molecular weight of about 5 kD to 500 kD, etc.
  • Biodegradable hydrogels may also be employed in the implants of the individual invention. Hydrogels are typically a copolymer material, characterized by the ability to imbibe a liquid. Exemplary biodegradable hydrogels which may be employed are described in Heller in: Hydrogels in Medicine and Pharmacy, N. A. Peppes ed., Vol. III, CRC Press, Boca Raton, Fla., 1987, pp 137-149.
  • compositions of the present disclosure containing an anti-C1q antibody of the present disclosure may be used (e.g., administered to an individual in need of treatment with anti-C1q antibody, such as a human individual) in accord with known methods, such as intravenous administration as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intracerobrospinal, intracranial, intraspinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes.
  • known methods such as intravenous administration as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intracerobrospinal, intracranial, intraspinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes.
  • Dosages and desired drug concentration of pharmaceutical compositions of the present disclosure may vary depending on the particular use envisioned. The determination of the appropriate dosage or route of administration is well within the skill of an ordinary artisan. Animal experiments provide reliable guidance for the determination of effective doses for human therapy. Interspecies scaling of effective doses can be performed following the principles described in Mordenti, J. and Chappell, W. “The Use of Interspecies Scaling in Toxicokinetics,” In Toxicokinetics and New Drug Development, Yacobi et al., Eds, Pergamon Press, New York 1989, pp. 42-46.
  • normal dosage amounts may vary from about 10 ng/kg up to about 100 mg/kg of an individual's and/or subject's body weight or more per day, depending upon the route of administration. In some embodiments, the dose amount is about 1 mg/kg/day to 10 mg/kg/day. For repeated administrations over several days or longer, depending on the severity of the disease, disorder, or condition to be treated, the treatment is sustained until a desired suppression of symptoms is achieved.
  • An exemplary dosing regimen may include administering an initial dose of an anti-C1q antibody, of about 2 mg/kg, followed by a weekly maintenance dose of about 1 mg/kg every other week.
  • Other dosage regimens may be useful, depending on the pattern of pharmacokinetic decay that the physician wishes to achieve. For example, dosing an individual from one to twenty-one times a week is contemplated herein. In certain embodiments, dosing ranging from about 3 ⁇ g/kg to about 2 mg/kg (such as about 3 ⁇ g/kg, about 10 ⁇ g/kg, about 30 ⁇ g/kg, about 100 ⁇ g/kg, about 300 ⁇ g/kg, about 1 mg/kg, or about 2 mg/kg) may be used.
  • dosing frequency is three times per day, twice per day, once per day, once every other day, once weekly, once every two weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every nine weeks, once every ten weeks, or once monthly, once every two months, once every three months, or longer. Progress of the therapy is easily monitored by conventional techniques and assays.
  • the dosing regimen, including the anti-C1q antibody administered, can vary over time independently of the dose used.
  • Dosages for a particular anti-C1q antibody may be determined empirically in individuals who have been given one or more administrations of the anti-C1q antibody. Individuals are given incremental doses of an anti-C1q antibody.
  • any clinical symptom of a neurodegenerative disorder e.g., Alzheimer's disease and Huntington's disease
  • inflammatory disorder e.g., IL-12, IL-12, or IL-12.
  • Administration of an anti-C1q antibody of the present disclosure can be continuous or intermittent, depending, for example, on the recipient's physiological condition, whether the purpose of the administration is therapeutic or prophylactic, and other factors known to skilled practitioners.
  • the administration of an anti-C1q antibody may be essentially continuous over a preselected period of time or may be in a series of spaced doses.
  • dosages and methods of delivery are provided in the literature; see, for example, U.S. Pat. Nos. 4,657,760; 5,206,344; or 5,225,212. It is within the scope of the invention that different formulations will be effective for different treatments and different disorders, and that administration intended to treat a specific organ or tissue may necessitate delivery in a manner different from that to another organ or tissue. Moreover, dosages may be administered by one or more separate administrations, or by continuous infusion. For repeated administrations over several days or longer, depending on the condition, the treatment is sustained until a desired suppression of disease symptoms occurs. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays.
  • anti-C1q antibodies and antigen-binding fragments thereof, which can bind to and neutralize a biologic activity of C1q.
  • These anti-C1 q antibodies are useful for preventing, reducing risk, or treating Alzheimer's disease and/or Huntington's disease.
  • anti-C1q antibodies of the present disclosure may be used for treating, preventing, or reducing risk of Alzheimer's disease and/or Huntington's disease in an individual.
  • the individual has Alzheimer's disease or Huntington's disease.
  • the individual is a human.
  • the Alzheimer's disease and/or Huntington's disease is associated with loss of nerve connections or synapses, including CF1-dependent synapse loss.
  • the synapse loss is dependent on the complement receptor 3 (CR3)C3 or complement receptor CR1 in some embodiments, the synapse loss is associated with pathological activity-dependent synaptic pruning.
  • synapses are phagocytosed by microglia.
  • Alzheimer s disease As disclosed herein, “Alzheimer's disease, ” “AD,” or “Alzheimer disease” may be used interchangeably and refer to a common form of dementia. Alzheimer s disease worsens as it progresses, and eventually leads to death. The cause and progression of Alzheimer's disease are not well understood. Alzheimer's disease is generally characterized by two types of brain lesions: senile plaques and neurofibrillary tangles.
  • Symptoms of Alzheimer's disease may include, without limitation, loss of cognitive function, short term memory loss, problems with the executive functions of attentiveness, planning, flexibility, and abstract thinking, impairments in semantic memory, confusion, irritability, aggression, mood swings, apathy, depression, impairment with learning and memory, difficulty with language, difficulty with executive functions, difficulty with perception (agnosia), difficulty with execution of movements (apraxia), long-term memory loss, loss of reading and writing skills, loss of complex motor functions, and loss of bodily functions.
  • the anti-C1q antibody may also be administered ex vivo in brain slice models of Alzheimer's disease.
  • the anti-C1q antibodies of the present disclosure may be used to treat, prevent, or improve one or more symptoms of Alzheimer's disease.
  • the present disclosure provides methods of treating, preventing, or improving one or more symptoms in subjects having Alzheimer's disease by administering an anti-C1q antibody of the present disclosure to, for example, inhibit the interaction between C1q and an autoantibody, such as an anti-ganglioside autoantibody, the interaction of C1q and C1r , and/or the interaction of C1q and C1s.
  • the anti-C1q antibody may also be administered ex vivo in brain slice models of Alzheimer's disease.
  • Huntington's disease and “HD” may be used interchangeably and refer to an inherited neurodegenerative genetic disorder that causes the progressive degeneration of nerve cells in the brain, affects muscle coordination, and leads to cognitive decline and psychiatric problems.
  • Huntington's disease is the most common genetic cause of abnormal involuntary writhing movements called chorea. Huntington's disease worsens as it progresses. Complications such as pneumonia, heart disease, and physical injury from falls reduce life expectancy to around twenty years from the point at which symptoms begin.
  • Huntington's disease is caused by inherited defect in a single gene (the Huntingtin gene).
  • Huntington's disease is an autosomal dominant disorder, which means that a person needs only one copy of the defective gene to develop Huntington's disease.
  • Huntington's disease usually causes movement, cognitive, and psychiatric disorders with a wide spectrum of signs and symptoms.
  • Symptoms of Huntington's disease may include, without limitation, problems with mood, problems with cognition, lack of coordination, involuntary jerking or writhing movements (chorea), involuntary, sustained contracture of muscles (dystonia), muscle rigidity, slow, uncoordinated fine movements, slow or abnormal eye movements, impaired gait, posture and balance, difficulty with the physical production of speech, difficulty swallowing, difficulty planning, organizing and prioritizing tasks, inability to start a task or conversation, lack of flexibility, the tendency to get stuck on a thought, behavior or action (perseveration), lack of impulse control that can result in outbursts, acting without thinking and sexual promiscuity, problems with spatial perception that can result in falls, clumsiness or accidents, lack of awareness of one's own behaviors and abilities, difficulty focusing on a task for long periods, slowness in processing thoughts or “finding” words, difficulty in learning new information
  • the anti-C1q antibodies of the present disclosure may be used to treat, prevent, or improve one or more symptoms of Huntington's disease.
  • the present disclosure provides methods of treating, preventing, or improving one or more symptoms in subjects having Huntington's disease by administering an anti-C1q antibody of the present disclosure to, for example, inhibit the interaction between C1q and an autoantibody, such as an anti-ganglioside autoantibody, the interaction of C1q and C1 r, and/or the interaction of C1q and C1s.
  • the anti-C1q antibody may also be administered ex vivo in brain slice models of Huntington's disease.
  • the antibodies of the present disclosure may be used, without limitation, in combination with any additional treatment for Alzheimer's disease and/or Huntington's disease.
  • an anti-C1q antibody of this disclosure is administered in therapeutically effective amounts in combination with a second anti-complement factor antibody (e.g., a neutralizing anti-complement factor antibody), such as an anti-C1s or anti-C1r antibody, or a second anti-C1q antibody.
  • a second anti-complement factor antibody e.g., a neutralizing anti-complement factor antibody
  • an anti-C1q antibody of this disclosure is administered in therapeutically effective amounts with a second and a third neutralizing anti-complement factor antibody, such as a second anti-C1q antibody, an anti-C1s antibody, and/or an anti-C1r antibody.
  • ADCC inhibitors may include, without limitation, soluble NK cell inhibitory receptors such as the killer cell Ig-like receptors (KIRs), which recognize HLA-A, HLA-B, or HLA-C and C-type lectin CD94/NKG2A heterodimers, which recognize HLA-E (see, e.g., Lopez-Botet M., T. Bellón, M. Llano, F. Navarro, P. Garcia & M. de Miguel. (2000), Paired inhibitory and triggering NK cell receptors for HLA class I molecules. Hum. Immunol.
  • KIRs killer cell Ig-like receptors
  • the antibodies of this disclosure are administered in combination with an inhibitor of the alternative pathway of complement activation.
  • inhibitors may include, without limitation, factor B blocking antibodies, factor D blocking antibodies, soluble, membrane-bound, tagged or fusion-protein forms of CD59, DAF, CR1, CR2, Crry or Comstatin-like peptides that block the cleavage of C3, non-peptide C3aR antagonists such as SB 290157, Cobra venom factor or non-specific complement inhibitors such as nafamostat mesilate (FUTHAN; FUT-175), aprotinin, K-76 monocarboxylic acid (MX-1) and heparin (see, e.g., T. E.
  • the antibodies of this disclosure are administered in combination with an inhibitor of the interaction between the autoantibody and its autoantigen.
  • inhibitors may include purified soluble forms of the autoantigen, or antigen mimetics such as peptide or RNA-derived mimotopes, including mimotopes of the AQP4 antigen.
  • inhibitors may include blocking agents that recognize the autoantigen and prevent binding of the autoantibody without triggering the classical complement pathway.
  • blocking agents may include, e.g., autoantigen-binding RNA aptamers or antibodies lacking functional C1q binding sites in their Fc domains (e.g., Fab fragments or antibody otherwise engineered not to bind C1q).
  • the anti-C1q antibodies of this disclosure also have diagnostic utility. This disclosure therefore provides for methods of using the anti-C1q antibodies of this disclosure, for diagnostic purposes, such as the detection of C1q in an individual or in tissue samples derived from an individual.
  • the individual is a human.
  • the individual is a human patient suffering from Alzheimer's disease or Huntington's disease.
  • the anti-C1q antibodies of this disclosure are used to detect synapses and synapse loss. For example, synapse loss may be measured in an individual suffering from Alzheimer's disease or Huntington's disease. The phenomenon of synapse loss in neurodegeneration is well understood in the art. See, e.g., U.S. Patent Publication Nos. 2012/0195880 and 2012/0328601.
  • the diagnostic methods involve the steps of administering an anti-C1q antibody of this disclosure to an individual and detecting the antibody bound to a synapse of the individual.
  • Antibody-binding to synapses may be quantified, for example, by non-invasive techniques such as positron emission tomography (PET), X-ray computed tomography, single-photon emission computed tomography (SPECT), computed tomography (CT), and computed axial tomography (CAT).
  • PET positron emission tomography
  • SPECT single-photon emission computed tomography
  • CT computed tomography
  • CAT computed axial tomography
  • the diagnostic methods involve detecting synapses in a biological sample, such as a biopsy specimen, a tissue, or a cell.
  • a biological sample such as a biopsy specimen, a tissue, or a cell.
  • An anti-C1q antibody is contacted with the biological sample and synapse-bound antibody is detected.
  • the detection method may involve quantification of the synapse-bound antibody.
  • Antibody detection in biological samples may occur with any method known in the art, including immunofluorescence microscopy, immunocytochemistry, immunohistochemistry, ELISA, FACS analysis or immunoprecipitation.
  • the quantification of synapse-bound antibodies provides a relative measure for the number of synapses present in the individual.
  • synapses are quantified repeatedly over a period of time.
  • the exact periodicity of synapse quantification depends on many factors, including the nature of the neurodegenerative disease (e.g., Alzheimer's disease and/or Huntington's disease), the stage of disease progression, treatment modalities and many other factors. Repeat measurements commonly reveal progressive synapse loss in individuals having a neurodegenerative disease (e.g., Alzheimer's disease and/or Huntington's disease).
  • relative synapse counts may be compared in populations of diseased individuals, and healthy control individuals at a single time point.
  • the treatment's efficacy can be assessed by comparing the rates of synapse loss in the treated individuals with the rates of synapse loss in a control group.
  • Control group members have received either no treatment or a control treatment, such as a placebo control.
  • kits containing anti-C1q antibodies of this disclosure for use in the methods of the present disclosure.
  • Kits of the invention may include one or more containers comprising a purified anti-C1q antibody of this disclosure.
  • the kits further include instructions for use in accordance with the methods of this disclosure.
  • these instructions comprise a description of administration of the anti-C1q antibody to treat or diagnose, e.g., Alzheimer's disease and/or Huntington's disease, according to any of the methods of this disclosure.
  • the instructions comprise a description of how to detect C1q, for example in an individual, in a tissue sample, or in a cell.
  • the kit may further comprise a description of selecting an individual suitable for treatment based on identifying whether that subject has the disease and the stage of the disease.
  • the instructions generally include information as to dosage, dosing schedule, and route of administration for the intended treatment.
  • the containers may be unit doses, bulk packages (e.g., multi-dose packages) or sub-unit doses.
  • Instructions supplied in the kits of the invention are typically written instructions on a label or package insert (e.g., a paper sheet included in the kit), but machine-readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable.
  • the label or package insert indicates that the composition is used for treating, e.g., Alzheimer's disease and Huntington's disease. Instructions may be provided for practicing any of the methods described herein.
  • kits of this invention are in suitable packaging.
  • suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like.
  • packages for use in combination with a specific device such as an inhaler, nasal administration device (e.g., an atomizer) or an infusion device such as a minipump.
  • a kit may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle).
  • the container may also have a sterile access port (e.g., the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle).
  • At least one active agent in the composition is an inhibitor of classical complement pathway.
  • the container may further comprise a second pharmaceutically active agent.
  • Kits may optionally provide additional components such as buffers and interpretive information.
  • the kit comprises a container and a label or package insert(s) on or associated with the container.
  • the anti-C1q antibody M1 was generated by Antibody Solutions Inc. (Sunnyvale Calif.) by immunizing C1q knockout mice with human C1q using standard mouse immunization and hybridoma screening technologies (Milstein, C (1999). Bioessays 21: 966-73; Mark Page, Robin Thorpe, The Protein Protocols Handbook 2002, Editors: John M. Walker. pp 1111-1113).
  • the anti-C1q antibodies 1C7, 2A1, 3A2, and 5A3 were generated at ImmunoPrecise Ltd (Victoria, BC Canada) by immunizing mice with human C1q protein purified from human plasma (Complement Technology Inc. Tyler Tex., Cat #A-099).
  • human C1q protein purified from human plasma Complement Technology Inc. Tyler Tex., Cat #A-099.
  • female BALB/c mice were injected intraperitoneal with 25 ⁇ g of protein in complete Freund's adjuvant (CFA) on Day 0 and boosts were done with 25 ⁇ g of C1q enzyme in incomplete Freund's adjuvant (IFA) on days 21, 42, 52, and a final intravenous boost on Day 63.
  • CFA complete Freund's adjuvant
  • IFA incomplete Freund's adjuvant
  • mice Four days following the final boost the mice were euthanized, spleens removed and splenocytes were fused with the myeloma cell line SP2/0. Fused cells were grown on hypoxanthine-aminopterin-thymidine (HAT) selective semisolid media for 10-12 days and the resulting hybridomas clones were transferred to 96-well tissue culture plates and grown in HAT medium until the antibody titer was high. The antibody-rich supernatants of the clones were isolated and tested in an ELISA assay for reactivity with C1q. Positive clones were isotyped and cultured for 32 days (post HAT selection) to identify stable expressing clones.
  • HAT hypoxanthine-aminopterin-thymidine
  • a hybridoma cell line producing the anti-C1q antibody M1 and referred to as mouse hybridoma C1q-M1 7788-1(M) 051613 was deposited with ATCC on 6/6/2013 having ATCC Accession Number PTA-120399.
  • M1 was shown to bind specifically to human and mouse C1q and to neutralize biological functions of C1q, such as complement mediated hemolysis (see, e.g., Example 3).
  • Anti-C1q antibodies 1C7, 2A1, 3A2, and 5A3 were screened for C1q-binding using standard ELISA protocols.
  • C1q-antibody interactions were measured using an OCTETM System according to standard protocols and manufacturer's instructions. Briefly, human and mouse C1q proteins were immobilized separately on a biosensor at three concentrations (3 nM, 1.0 nM, and 0.33 nM). Next, the anti-C1q antibody M1 was injected onto the C1q-coated biosensor at a concentration of 2.0 ⁇ g/ml and the association constants (k on ) and dissociation constants (k off ) for anti-C1q antibodies M1 and 4A4B11 were measured.
  • anti-C1q antibody M1 was shown to bind both human and mouse C1q proteins with very high affinities (K D ⁇ 10 ⁇ 10 M).
  • the reference antibody 4A4B11 was found to bind to human C1q, whereas binding to mouse C1q was undetectable.
  • the affinities of M1 and 4A4B11 for human C1q were on the same order of magnitude (i.e., in the double-digit picomolar range; K D ⁇ 10-30 pM)
  • the affinity of M1 for mouse C1q was determined to be about four orders of magnitude higher (K D ⁇ 30 pM) than that the affinity of 4A4B 11 for mouse C1q (K D ⁇ 40 nM).
  • Blocking experiments were performed to determine whether the anti-C1q antibodies M1 and 4A4B11 bind to the same or overlapping epitopes of human C1q or whether M1 and 4A4B11 bind to separate C1q epitopes.
  • M1 was coated on a biosensor chip (BIACORETM) and subsequently contacted with a combination of human C1q and M1, a combination of human C1q and 4A4B11, or human C1q alone.
  • C1q-binding to M1 was followed for 10 min and dissociation of M1-C1q complexes was subsequently followed for 20 min. Relative binding signals were recorded at the end of the association and dissociation periods.
  • Table C shows the results of these experiments.
  • Anti-C1q antibodies were tested in human and rodent hemolytic assays (CH50) for their ability to neutralize C1q and block its activation of the downstream complement cascade.
  • CH50 assays were conducted essentially as described in Current Protocols in Immunology (1994) Supplement 9 Unit 13.1.
  • 5 microliters ( ⁇ l) of human serum (Cedarlane, Burlington, N.C.), 0.625 ⁇ l of Wistar rat serum, or 2.5 ⁇ l of C57B1/6 mouse serum was diluted to 50 ⁇ l of GVB buffer (Cedarlane, Burlington, N.C.) and added to 50 ⁇ l of the monoclonal antibodies (1 ⁇ g) diluted in GVB buffer.
  • the antibody:serum mixture was pre-incubated for 30 minutes on ice and then added to 100 ⁇ l of EA cells (2 ⁇ 10 8 /ml) for rat and human assays, and 4 ⁇ 10 7 /ml for mouse assays.
  • the EA cells were generated exactly as specified in Current Protocols using Sheeps blood in Alsever's (Cedarlane Cat #CL2581) and hemolysin (Cedalane Cat #CL9000).
  • the EA cells, serum and antibody mixture was incubated for 30 minutes at 37° C. and then placed on ice. Next 1.2 ml of 0.15 M NaCl was added to the mixture and the OD 412 of the sample was read in a spectrophotometer to determine the amount of cell lysis.
  • the percent inhibition of the test antibodies was determined relative to a control mouse IgG1 antibody (Abcam ab 18447).
  • a modified CH50 assay (also referred to as C1F hemolysis assay) was performed that provided limiting quantities of the C1 complex from human serum to provide greater sensitivity for assessing C1 activity and potential C1 inhibition.
  • the assay was conducted as follows. First, 3 ⁇ 10 7 sheep red blood cells (RBC) were incubated with anti-sheep RBC IgM antibody to generate activated erythrocytes (EA cells). The EA cells were then incubated with purified C4b protein to create EAC4b cells.
  • RBC sheep red blood cells
  • EA cells activated erythrocytes
  • EAC4b cells were subsequently incubated with diluted (1:1000-1:10000) normal human serum (NHS) that was pre-incubated with or without anti-C1q and control mouse IgG antibodies, to provide a limiting quantity of human C1.
  • NHS normal human serum
  • the resulting EAC14 cells were incubated with purified human C2 protein to generate EAC14b2a cells.
  • guinea pig serum was added in an EDTA buffer and incubated at 37° C. for 30 minutes. Cell lysis was measured in a spectrophotometer at a wavelength of 450 nm.
  • C1q-binding antibodies (1C7, 2A1, 3A2, and 5A3) were tested in the human CH50 assay at a single concentration (1 ⁇ g) ( FIG. 2 ). All four antibodies were found to inhibit hemolysis.
  • the anti-C1q antibody 1C7 inhibited hemolysis at greater than 90%, 2A1 inhibited hemolysis at greater than 40%, 3A2 inhibited hemolysis at greater than 60%, and 5A3 inhibited hemolysis at greater than 50%.
  • anti-C1q antibodies 1C7 and 3A2 were tested in the human CH50 hemolysis assay in a dose-response format ( FIG. 3 ).
  • Anti-C1q antibody 4A4B11 was used as a reference. Both 1C7 and 3A2 antibodies inhibited CH50 hemolysis in a dose-dependent manner. Approximately 100 ng of the 1C7 antibody and approximately 200 ng of the 3A2 were required to inhibit 50% of the hemolysis observed ( FIG. 3 ).
  • Anti-C1q antibody M1 was tested for its C1q neutralizing activity in human, mouse, and rat CH50 assays ( FIG. 4A-C ). Testing was conducted in dose-response formats. Anti-C1q antibody 4A4B11 was used as a reference. M1 was demonstrated to neutralize C1q activity in human, mouse, and rat CH50 hemolysis assays in a dose-dependent manner ( FIG. 4A -4C). By contrast, 4A4B11 was found to neutralize C1q activity only in the human CH50 assay, whereas the reference antibody was inactive in the mouse and rat CH50 hemolysis assays (up to 2 ⁇ g).
  • the inhibition of the interaction between the C1Q protein and the antibodies 4A4B11 (ANN-001) and M1 (ANN-005) by unstructured peptides generated by proteolysis of the C1q antigen was evaluated. If the peptides generated by complete proteolysis of the antigen are able to inhibit the binding of the antigen on the antibody, the interaction is not based on conformation, and the epitope is linear. If the peptides generated by complete proteolysis of the antigen are unable to inhibit the binding of the antigen on the antibodies 4A4B11 and Ml, the conformation is necessary for interaction.
  • the antibody/antigen complexes were incubated with deuterated cross-linkers and subjected to multi-enzymatic proteolytic cleavage. After enrichment of the cross-linked peptides, the samples were analyzed by high resolution mass spectrometry (nLC-Orbitrap MS) and the data generated analyzed using XQuest software.
  • antibody 4A4B11 (ANN-001) binds to an epitope that includes amino acids S202 and K219 of human C1QA and Y225 of human C1QC
  • antibody M1 (ANN-005) binds to an epitope that includes amino acid K219 of human C1QA and S185 of human C1QC. See the amino acid sequence alignment of human and mouse C1qA and C1qC as shown below.
  • the C1q/antibody complexes were generated by mixing equimolar solutions of C1q antigen and antibody (4 ⁇ M in 5 ⁇ l each).
  • One ⁇ l of the mixture obtained was mixed with 1 ⁇ l of a matrix composed of a re-crystallized sinapinic acid matrix (10 mg/ml) in acetonitrile/water (1:1, v/v), TFA 0.1% (K200 MALDI Kit).
  • 1 ⁇ l of each sample was spotted on the MALDI plate (SCOUT 384). After crystallization at room temperature, the plate was introduced in the MALDI mass spectrometer and analyzed immediately. The analysis has been repeated in triplicate.
  • FIG. 5 shows the presence of the antigen, antibody and antigen/antibody complexes for C1q/4A4B 11 ( FIG. 5A ) and C1q/M1 ( FIG. 5B ). Peaks are present at the predicted molecular weights of monomeric antibody ( ⁇ 150 kDa) and C1q monomer ( ⁇ 460 kDa) and there is a 1:1 complex of antibody:antigen present at ⁇ 615 kDa.
  • the C1q antigen was digested with immobilized pepsin. 25 ⁇ l of the antigen with a concentration of 10 ⁇ M were mixed with immobilized pepsin 5 ⁇ M and incubate at room temperature for 30 minutes. After the incubation time the sample was centrifuged and the supernatant was pipetted. The completion of the proteolysis was controlled by High-Mass MALDI mass spectrometry in linear mode. The pepsin proteolysis was optimized in order to obtain a large amount of peptide in the 1000-3500 Da range.
  • the MALDI ToF MS analysis was performed using CovalX's HM3 interaction module with a standard nitrogen laser and focusing on different mass ranges from 0 to 2000 kDa.
  • Mass Spectrometer Linear and Positive mode; Ion Source 1: 20 kV; Ion Source 2: 17 kV; Pulse Ion Extraction: 400 ns; for HM3: Gain Voltage: 3.14 kV; Gain Voltage: 3.14 kV; Acceleration Voltage: 20 kV.
  • DSS-H12 DiSuccinimidylSuberate H12
  • DSS-D12 DiSuccinimidylSuberate D12
  • the 2 mg prepared were mixed with 1 ml of DMF in order to obtain a 2 mg/ml solution of DSS H12/D12.
  • 10 ⁇ l of the antibody/antigen mix prepared previously were mixed with 1 ⁇ l of the solution of cross-linker d0/d12 prepared (2 mg/ml).
  • the solution was incubated 180 minutes at room temperature in order to achieve the cross-linking reaction. In order to facilitate the proteolysis, it was necessary to reduce the disulfide bound present in this protein.
  • the cross-linked sample was mixed with 20 ⁇ l of ammonium bicarbonate (25 mM, pH 8.3). After mixing 2.5 ⁇ l of DTT (500 mM) is added to the solution. The mixture was then incubated 1 hour at 55° C. After incubation, 2.5 ⁇ l of iodioacetamide (1 M) was added before 1 hour of incubation at room temperature in a dark room. After incubation, the solution was diluted 1 ⁇ 5 by adding 120 ⁇ l of the buffer used for the proteolysis. 145 ⁇ l of the reduced/alkyled cross-linked sample was mixed with 2 ⁇ l of trypsin (Sigma, T6567). The proteolytic mixture was incubated overnight at 37° C.
  • the buffer of proteolysis was Tris-HCL 100 mM, CaCl 2 10 mM, pH7.8.
  • the 145 ⁇ l of the reduced/alkyled cross-linked complex was mixed with 2 ⁇ l of ⁇ -chymotrypsin 200 ⁇ M and incubated overnight at 30° C.
  • an nLC in combination with Orbitrap mass spectrometry were used.
  • the cross-linker peptides were analyzed using Xquest version 2.0 and stavrox software. The peptides identified and cross-linked amino acids are indicated in Table D below.
  • mice were perfused with 4% PFA and their brains extracted and processed for immunohistochemistry (IHC). Briefly, the tissue was postfixed for 2 hr in PFA before washing in PBS and incubating for 48 hr in 30% sucrose in PBS. The brains were subsequently frozen in OCT and 14 mm cryosections were cut.
  • IHC immunohistochemistry
  • Brain sections were dried at 37° C. for 30 minutes, washed in PBS, and then incubated with blocking solution for 1 hour (hr) at room temperature (150 mM NaCl 50 mM Tris base, 5% BSA, 100 mM L-lysine, 0.2% triton, 0.04% sodium azide). Sections were subsequently incubated with relevant primary antibody diluted in blocking solution overnight at 4° C.
  • C1q staining undiluted rabbit anti-C1q antibody (Stephen AH et al., J Neurosci. 2013 Aug. 14; 33(33):13460-74) was used.
  • C3 staining goat anti-mouse C3 (Cappel 55730) was used at a dilution of 1/200.
  • Synapses were labeled with rat anti-mouse PSD-95 (Millipore MAB1596) used at a dilution of 1/100 and with rabbit anti-mouse synapsin (Synaptic systems 106103) used at a dilution of 1/500.
  • Sections were then washed in PBS and incubated with fluorescently tagged secondary antibodies diluted in blocking buffer for 2 hr at room temperature.
  • For C3 staining donkey anti-goat Alexa Fluor 488 (Life Technologies A11055) and donkey anti-goat Alexa Fluor 594 (Life Technologies A11058) secondary antibodies were used; for PSD-95 staining, donkey anti-mouse Alexa Fluor 488 (Life Technologies A21202) secondary antibody was used; for synapsin staining, donkey anti-rabbit Alexa Fluor 594 (Life Technologies A21207) secondary antibody was used; and for C1q staining, donkey anti-rabbit Alexa Fluor 488 (Life Technologies A21206) secondary antibody was used. After washing in PBS, sections were mounted in vectashield (Vector Labs H-1400).
  • Sections were imaged on a Zeiss LSM 700 confocal microscope using a 63 ⁇ oil objective and a 1AU pinhole. Single plane images from each channel were captured in relevant brain regions. Image J software was then used to threshold the images and quantify the number of co-localized puncta.
  • An acute in vivo model of A ⁇ synaptotoxicity in Alzheimer's disease was generated by injecting 5 ng A ⁇ oligomers or saline (1 ⁇ l vol.) at 0.5 ⁇ l/min via intracerebroventricular (ICV) injection using a Hamilton syringe into anesthetized wild-type (WT) mice following the coordinates for left ventral placement (bregma-0.4 mm, 1.0 mm lateral to midline, and 2.5 mm below dura at 0° angle).
  • C1q neutralizing antibody trial 5 ⁇ g C1q neutralizing antibody or control IgG (1 ⁇ l vol.) was injected to the left ventricle of C57BL/6J mice, followed by 5 ng A ⁇ oligomers or saline (1 ⁇ l vol.).
  • Adult (2-3 mo) wild-type (WT) mice C57BL/6J; JAX 000664) or C1qA knockout (KO) mice (Botto et al., 1998) were then given an intraperitoneal (IP) injection of C1q neutralizing antibody or control IgG (20 mg/kg), and were allowed to wake and resume normal activities.
  • WT wild-type mice
  • KO C1qA knockout mice
  • mice were sacrificed 18 hours post injection.
  • IP intraperitoneal
  • zQ175 is a knock-in model of Huntington's disease in which 188 CAG repeats have been inserted into the mouse Huntingtin gene alongside the human polyproline region. It was generated from a spontaneous expansion of the CAG repeat region in a litter of CAG 140 mice. These mice phenocopy many aspects of the human disease with robust motor and cognitive deficits starting around 30 weeks of age, as shown by a reduced performance on the rotarod and in a procedural two choice-swim test. They also show specific striatal atrophy, an early event in human Huntington's disease pathology with a 21% loss of volume at 30 weeks.
  • MSNs Striatal medium spiny neurons
  • mHTT insult Striatal medium spiny neurons
  • dramatic loss of this neuronal population in Huntington's disease patients is a hallmark of the disease.
  • MSNs in zQ175 mice show hyper-excitability from as early as 12 weeks, followed by progressive loss of corticostriatal transmission (Heikkinen et al., 2012; Menalled et al., 2012).
  • the C1q blocking antibody M1 was generated and characterized as described in Examples 1-4 above.
  • FIG. 8A shows that A ⁇ oligomers induce much higher levels of C1q deposition in the brain, as compared to the A ⁇ monomers.
  • FIG. 8B shows that C1q is co-localized at synapses with PSD-95. Moreover, consistent with the results in FIG. 8A , A ⁇ oligomers induced a greater amount of C1q co-localization with PSD-95 at synapses ( FIG. 8B ).
  • synapse number was evaluated in wild-type (WT) mice and C1q knockout (KO) mice injected with A ⁇ oligomers.
  • Intracerebroventricular (ICV) injection of either soluble A ⁇ oligomers or A ⁇ monomers was performed in healthy adult (2-3 mo) WT and C1q KO mice.
  • brains were harvested for immunohistochemistry (IHC) using synapsin as pre-synaptic marker and PSD95 as a post-synaptic marker.
  • Synapse number was quantified by measuring the number of co-localized synapsin and PSD-95 puncta. The results in FIG.
  • FIG. 9 shows that a significant loss of structural synapses in the CA1 region of the hippocampus occurs A ⁇ oligomer-treated WT mice ( FIGS. 9A and 9B ), but does not occur in C1q KO mice ( FIG. 9C ). These results demonstrate that C1q deficiency prevents the loss of synaptic puncta, which is typically seen in wild-type mice after A ⁇ oligomer treatment.
  • FIG. 10 shows that the deposition of C3 induction by A ⁇ oligomer treatment in wild-type (WT) mice is absent in C1q KO mice treated A ⁇ oligomers.
  • the ability of the anti-C1q antibody M1 to suppress synapse loss in an in vivo mouse model of Alzheimer's disease (AD) was tested using the acute A ⁇ oligomer model of AD in which A ⁇ oligomers are injected directly into the ventricles of the brain.
  • Two sets of mice were given an initial IP injection and ICV injection of a blocking anti-C1q antibody and a mouse IgG1 control alongside an ICV injection of A ⁇ oligomers.
  • One set of mice was sacrificed after 18 hr to determine whether the antibody was capable of reducing A ⁇ oligomer-induced C3 levels (a downstream complement component).
  • the other set of mice received a further IP injection of the blocking anti-C1q antibody 48 hr after the initial surgery, and sacrificed at 72 hr to assess levels of synaptic markers.
  • FIG. 11 shows that in wild-type (WT) mice co-injected with the C1q blocking antibody and A ⁇ oligomers there is a significant reduction in C3 staining ( FIG. 11A ) in the brain and more synaptic puncta ( FIG. 11B ), as compared to mice co-injected with the IgG1 control and A ⁇ oligomers.
  • the efficacy of the C1q antibody M1 was also tested in an in vivo mouse model of Huntington's disease (zQ125 transgenic mice).
  • zQ125 transgenic mice received two IP injections over a 48 hr period of 20 mg/kg of the M1 antibody or of a mouse IgG control. Consistent with the results of Example 8 above using a mouse model of Alzheimer's disease, there was less C3 deposition in the disease-affected regions of the zQ175 mice injected with the C1q blocking antibody, as compared to mice injected with the IgG control or control mice that did not receive an injection ( FIG. 12 ).
  • the hybridoma cell line producing the M1 antibody (mouse hybridoma C1qM1 7788-1(M) 051613) has been deposited with ATCC under conditions that assure that access to the culture will be available during pendency of the patent application and for a period of 30 years, or 5 years after the most recent request, or for the effective life of the patent, whichever is longer. A deposit will be replaced if the deposit becomes nonviable during that period. The deposit is available as required by foreign patent laws in countries wherein counterparts of the subject application, or its progeny are filed. However, it should be understood that the availability of the deposit does not constitute a license to practice the subject invention in derogation of patent rights granted by governmental action.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Diabetes (AREA)
  • Neurology (AREA)
  • Hematology (AREA)
  • Neurosurgery (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Ophthalmology & Optometry (AREA)
  • Obesity (AREA)
  • Pulmonology (AREA)
  • Cell Biology (AREA)
  • Pathology (AREA)
  • Endocrinology (AREA)
  • Psychology (AREA)
  • Rheumatology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Microbiology (AREA)
  • Virology (AREA)
  • Zoology (AREA)
US14/988,387 2013-07-09 2016-01-05 Methods of treatment for alzheimer's disease and huntington's disease Abandoned US20160159890A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US14/988,387 US20160159890A1 (en) 2013-07-09 2016-01-05 Methods of treatment for alzheimer's disease and huntington's disease

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201361844369P 2013-07-09 2013-07-09
US201361871813P 2013-08-29 2013-08-29
PCT/US2014/046045 WO2015006507A1 (en) 2013-07-09 2014-07-09 Methods of treatment for alzheimer's disease and huntington's disease
US14/988,387 US20160159890A1 (en) 2013-07-09 2016-01-05 Methods of treatment for alzheimer's disease and huntington's disease

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2014/046045 Continuation WO2015006507A1 (en) 2013-07-09 2014-07-09 Methods of treatment for alzheimer's disease and huntington's disease

Publications (1)

Publication Number Publication Date
US20160159890A1 true US20160159890A1 (en) 2016-06-09

Family

ID=52280583

Family Applications (7)

Application Number Title Priority Date Filing Date
US14/988,387 Abandoned US20160159890A1 (en) 2013-07-09 2016-01-05 Methods of treatment for alzheimer's disease and huntington's disease
US14/989,038 Active US9708394B2 (en) 2013-07-09 2016-01-06 Anti-complement factor C1q antibodies and uses thereof
US15/491,574 Active US10227398B2 (en) 2013-07-09 2017-04-19 Anti-complement factor C1q antibodies and uses thereof
US16/239,685 Active US10590190B2 (en) 2013-07-09 2019-01-04 Anti-complement factor C1q antibodies and uses thereof
US16/784,020 Active US10927167B2 (en) 2013-07-09 2020-02-06 Anti-complement factor C1Q antibodies and uses thereof
US17/172,807 Active 2035-02-21 US11649279B2 (en) 2013-07-09 2021-02-10 Anti-complement factor C1Q antibodies and uses thereof
US18/110,574 Pending US20240083987A1 (en) 2013-07-09 2023-02-16 Anti-complement factor c1q antibodies and uses thereof

Family Applications After (6)

Application Number Title Priority Date Filing Date
US14/989,038 Active US9708394B2 (en) 2013-07-09 2016-01-06 Anti-complement factor C1q antibodies and uses thereof
US15/491,574 Active US10227398B2 (en) 2013-07-09 2017-04-19 Anti-complement factor C1q antibodies and uses thereof
US16/239,685 Active US10590190B2 (en) 2013-07-09 2019-01-04 Anti-complement factor C1q antibodies and uses thereof
US16/784,020 Active US10927167B2 (en) 2013-07-09 2020-02-06 Anti-complement factor C1Q antibodies and uses thereof
US17/172,807 Active 2035-02-21 US11649279B2 (en) 2013-07-09 2021-02-10 Anti-complement factor C1Q antibodies and uses thereof
US18/110,574 Pending US20240083987A1 (en) 2013-07-09 2023-02-16 Anti-complement factor c1q antibodies and uses thereof

Country Status (23)

Country Link
US (7) US20160159890A1 (enExample)
EP (3) EP3019523A4 (enExample)
JP (2) JP6462680B2 (enExample)
KR (1) KR102321320B1 (enExample)
CN (1) CN105682740B (enExample)
AU (1) AU2014287221C1 (enExample)
BR (1) BR112016000106B1 (enExample)
CA (1) CA2916521C (enExample)
DK (1) DK3019240T3 (enExample)
EA (1) EA037325B1 (enExample)
ES (1) ES2978917T3 (enExample)
FI (1) FI3019240T3 (enExample)
HU (1) HUE066722T2 (enExample)
IL (1) IL243506B (enExample)
MX (2) MX375500B (enExample)
MY (1) MY175896A (enExample)
NZ (1) NZ715455A (enExample)
PL (1) PL3019240T3 (enExample)
PT (1) PT3019240T (enExample)
SG (3) SG11201510316UA (enExample)
SI (1) SI3019240T1 (enExample)
WO (2) WO2015006507A1 (enExample)
ZA (1) ZA201600157B (enExample)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160355574A1 (en) * 2014-11-05 2016-12-08 Arnon Rosenthal Humanized anti-complement factor c1q antibodies and uses thereof
WO2018071676A1 (en) * 2016-10-12 2018-04-19 Bioverativ Usa Inc. Anti-c1s antibodies and methods of use thereof
US10227398B2 (en) 2013-07-09 2019-03-12 Annexon, Inc. Anti-complement factor C1q antibodies and uses thereof
US10450382B2 (en) 2012-11-02 2019-10-22 Bioverativ Usa Inc. Anti-complement C1s antibodies
US10457745B2 (en) 2012-10-25 2019-10-29 Bioverativ Usa Inc. Anti-complement C1s antibodies
US10723788B2 (en) 2015-11-24 2020-07-28 Annexon, Inc. Anti-complement factor C1q Fab fragments and uses thereof
US10729767B2 (en) 2015-04-06 2020-08-04 Bioverativ Usa Inc. Humanized anti-C1s antibodies and methods of inhibiting C1s cleavage
WO2021226513A1 (en) 2020-05-08 2021-11-11 President And Fellows Of Harvard College Methods for treating inflammatory and autoimmune disorders
EP4045084A4 (en) * 2019-10-17 2023-05-31 Annexon, Inc. COMPOSITIONS AND METHODS FOR TREATING BLOOD DISORDERS
WO2023130001A1 (en) * 2021-12-30 2023-07-06 Annexon, Inc. Compositions and methods for treating huntington's disease
CN120685919A (zh) * 2025-08-22 2025-09-23 中国科学院长春应用化学研究所 一种C1q补体蛋白在制备检测Aβ42蛋白的元件中的应用、一种检测Aβ42蛋白的芯片及其应用
US12486322B2 (en) 2021-12-13 2025-12-02 Annexon, Inc. Anti-complement factor C1q antibodies with single binding arms and uses thereof

Families Citing this family (31)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8148330B2 (en) 2005-12-09 2012-04-03 The Board Of Trustees Of The Leland Stanford Junior University Modulation of synaptic maintenance
US9480658B2 (en) 2005-12-09 2016-11-01 The Board Of Trustees Of The Leland Stanford Junior University Modulation of synaptic maintenance
EP2996722A4 (en) * 2013-05-15 2017-01-11 Annexon, Inc. Methods of treatment for guillain-barre syndrome
LT3280441T (lt) 2015-04-07 2021-11-25 Alector Llc Anti-sortilino antikūnai ir jų naudojimo būdai
US20190161535A1 (en) * 2016-05-09 2019-05-30 Annexon, Inc. Compositions and methods for treating spinal muscular atrophy
WO2018017711A1 (en) * 2016-07-19 2018-01-25 Annexon, Inc. Compositions and methods for treating frontotemporal dementia
EP3515932B1 (en) 2016-09-19 2023-11-22 F. Hoffmann-La Roche AG Complement factor based affinity chromatography
CN106699883B (zh) * 2016-12-19 2020-08-07 中国兽医药品监察所 豚鼠补体C1q-B单克隆抗体60G4的制备与应用
CN106822863A (zh) * 2017-01-04 2017-06-13 国家纳米科学中心 多肽‑抗体免疫偶联物及其制备方法
CN109843927B (zh) * 2017-03-06 2022-06-21 江苏恒瑞医药股份有限公司 抗b7-h3抗体、其抗原结合片段及其医药用途
CN111386039B (zh) * 2017-09-29 2023-02-28 瑞泽恩制药公司 经基因修饰的啮齿动物基因组
EP4640703A2 (en) 2017-11-14 2025-10-29 Chugai Seiyaku Kabushiki Kaisha Anti-c1s antibodies and methods of use
WO2019137922A1 (en) * 2018-01-09 2019-07-18 Leibniz-Institut Für Naturstoff-Forschung Und Infektionsbiologie Modulators of c1q, in particular of the interaction of apoe with c1q, and uses of the modulators in the therapy of neuronal diseases and inflammation
EP4257602A3 (en) * 2018-06-11 2023-12-27 Aarhus Universitet Single domain antibodies for complement regulation
PE20210186A1 (es) 2018-07-13 2021-02-02 Alector Llc Anticuerpos anti-sortilina y metodos para su uso
US12422440B2 (en) 2018-10-05 2025-09-23 Seattle Children's Hospital Newborn screening for primary immunodeficiencies, cystinosis, and Wilson disease
JP7499760B2 (ja) * 2018-11-02 2024-06-14 アネクソン，インコーポレーテッド 脳損傷を治療するための組成物及び方法
BR112021021689A2 (pt) 2019-05-15 2022-03-22 Chugai Pharmaceutical Co Ltd Molécula de ligação a antígeno, composição farmacêutica, e método
BR112022006014A2 (pt) * 2019-10-16 2022-07-12 Chugai Pharmaceutical Co Ltd Anticorpo, composição farmacêutica e método
US11940448B2 (en) 2020-03-31 2024-03-26 Seattle Children's Hospital Proteomic screening for lysosomal storage diseases
CA3177879A1 (en) * 2020-05-05 2021-12-02 Jeanne T. PAZ Compositions and methods for treating epilepsy
WO2021250323A1 (en) * 2020-06-09 2021-12-16 Helsingin Yliopisto Anti-cox-2 autoantibody as a diagnostic marker, and methods, kits and uses related thereto
KR20230041758A (ko) * 2020-07-20 2023-03-24 애넥슨, 인코포레이티드 보체 인자의 저해제 및 이의 용도
AU2021361067A1 (en) * 2020-10-16 2023-06-01 Annexon, Inc. Compositions and methods for treating blood disorders
CA3200976A1 (en) * 2020-12-04 2022-06-09 Anita GROVER Compositions and methods for treating ocular diseases
GB202113673D0 (en) * 2021-09-24 2021-11-10 Reflection Therapeutics Ltd Targeted cell therapies
JP2024539141A (ja) * 2021-10-21 2024-10-28 アネクソン，インコーポレーテッド 筋ジストロフィーを治療するための組成物及び方法
EP4453571A1 (en) * 2021-12-29 2024-10-30 Curateq Biologics Private Limited C1q binding assay
EP4598575A1 (en) * 2022-10-07 2025-08-13 Annexon, Inc. Formulations for anti-c1q antibodies
WO2024154122A1 (en) 2023-01-18 2024-07-25 Gilboa Therapeutics LTD Immune cells expressing a complement receptor and uses thereof
CN119326377B (zh) * 2024-10-15 2025-07-22 复旦大学 一种基于影像的认知水平评估方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150259437A1 (en) * 2012-10-25 2015-09-17 True North Therapeutics, Inc. Anti-complement c1s antibodies and uses thereof

Family Cites Families (100)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4657760A (en) 1979-03-20 1987-04-14 Ortho Pharmaceutical Corporation Methods and compositions using monoclonal antibody to human T cells
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US4595654A (en) 1983-11-07 1986-06-17 Immunomedics Inc. Method for detecting immune complexes in serum
US5206344A (en) 1985-06-26 1993-04-27 Cetus Oncology Corporation Interleukin-2 muteins and polymer conjugation thereof
US4676980A (en) 1985-09-23 1987-06-30 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Target specific cross-linked heteroantibodies
GB8601597D0 (en) 1986-01-23 1986-02-26 Wilson R H Nucleotide sequences
US6548640B1 (en) 1986-03-27 2003-04-15 Btg International Limited Altered antibodies
US5567610A (en) 1986-09-04 1996-10-22 Bioinvent International Ab Method of producing human monoclonal antibodies and kit therefor
DE3850542T2 (de) 1987-09-23 1994-11-24 Bristol Myers Squibb Co Antikörper-Heterokonjugate zur Töting von HIV-infizierten Zellen.
GB8823869D0 (en) 1988-10-12 1988-11-16 Medical Res Council Production of antibodies
US5175384A (en) 1988-12-05 1992-12-29 Genpharm International Transgenic mice depleted in mature t-cells and methods for making transgenic mice
DE3920358A1 (de) 1989-06-22 1991-01-17 Behringwerke Ag Bispezifische und oligospezifische, mono- und oligovalente antikoerperkonstrukte, ihre herstellung und verwendung
ATE144793T1 (de) 1989-06-29 1996-11-15 Medarex Inc Bispezifische reagenzien für die aids-therapie
US5225212A (en) 1989-10-20 1993-07-06 Liposome Technology, Inc. Microreservoir liposome composition and method
US5959177A (en) 1989-10-27 1999-09-28 The Scripps Research Institute Transgenic plants expressing assembled secretory antibodies
US6075181A (en) 1990-01-12 2000-06-13 Abgenix, Inc. Human antibodies derived from immunized xenomice
DE69133566T2 (de) 1990-01-12 2007-12-06 Amgen Fremont Inc. Bildung von xenogenen Antikörpern
US6150584A (en) 1990-01-12 2000-11-21 Abgenix, Inc. Human antibodies derived from immunized xenomice
US5229275A (en) 1990-04-26 1993-07-20 Akzo N.V. In-vitro method for producing antigen-specific human monoclonal antibodies
JPH06500011A (ja) 1990-06-29 1994-01-06 ラージ　スケール　バイオロジー　コーポレイション 形質転換された微生物によるメラニンの製造
US5633425A (en) 1990-08-29 1997-05-27 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
KR100272077B1 (ko) 1990-08-29 2000-11-15 젠팜인터내셔날,인코포레이티드 이종 항체를 생산할 수 있는 전이유전자를 가진 인간이외의 동물
US5625126A (en) 1990-08-29 1997-04-29 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US5661016A (en) 1990-08-29 1997-08-26 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5545806A (en) 1990-08-29 1996-08-13 Genpharm International, Inc. Ransgenic non-human animals for producing heterologous antibodies
US5571894A (en) 1991-02-05 1996-11-05 Ciba-Geigy Corporation Recombinant antibodies specific for a growth factor receptor
WO1992020373A1 (en) 1991-05-14 1992-11-26 Repligen Corporation Heteroconjugate antibodies for treatment of hiv infection
WO1993006217A1 (en) 1991-09-19 1993-04-01 Genentech, Inc. EXPRESSION IN E. COLI OF ANTIBODY FRAGMENTS HAVING AT LEAST A CYSTEINE PRESENT AS A FREE THIOL, USE FOR THE PRODUCTION OF BIFUNCTIONAL F(ab')2 ANTIBODIES
US5565332A (en) 1991-09-23 1996-10-15 Medical Research Council Production of chimeric antibodies - a combinatorial approach
FI941572L (fi) 1991-10-07 1994-05-27 Oncologix Inc Anti-erbB-2-monoklonaalisten vasta-aineiden yhdistelmä ja käyttömenetelmä
WO1993008829A1 (en) 1991-11-04 1993-05-13 The Regents Of The University Of California Compositions that mediate killing of hiv-infected cells
AU3178993A (en) 1991-11-25 1993-06-28 Enzon, Inc. Multivalent antigen-binding proteins
ATE419355T1 (de) 1992-02-06 2009-01-15 Novartis Vaccines & Diagnostic Marker für krebs und biosynthetisches bindeprotein dafür
US5573905A (en) 1992-03-30 1996-11-12 The Scripps Research Institute Encoded combinatorial chemical libraries
ATE149570T1 (de) 1992-08-17 1997-03-15 Genentech Inc Bispezifische immunoadhesine
US5789199A (en) 1994-11-03 1998-08-04 Genentech, Inc. Process for bacterial production of polypeptides
US5840523A (en) 1995-03-01 1998-11-24 Genetech, Inc. Methods and compositions for secretion of heterologous polypeptides
US5731168A (en) 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
US5641870A (en) 1995-04-20 1997-06-24 Genentech, Inc. Low pH hydrophobic interaction chromatography for antibody purification
US5869046A (en) 1995-04-14 1999-02-09 Genentech, Inc. Altered polypeptides with increased half-life
US5739277A (en) 1995-04-14 1998-04-14 Genentech Inc. Altered polypeptides with increased half-life
JP4312259B2 (ja) 1995-04-27 2009-08-12 アムジェン フレモント インク． 免疫したゼノマウス（ＸｅｎｏＭｏｕｓｅ）に由来するヒト抗体
EP0823941A4 (en) 1995-04-28 2001-09-19 Abgenix Inc HUMAN ANTIBODIES DERIVED FROM IMMUNIZED XENO MOUSES
US6717031B2 (en) 1995-06-07 2004-04-06 Kate Dora Games Method for selecting a transgenic mouse model of alzheimer's disease
US20020058311A1 (en) 1995-06-13 2002-05-16 Browne Michael Jospeh Chimeric leptin fused to immunoglobulin domain and use
DE19544393A1 (de) 1995-11-15 1997-05-22 Hoechst Schering Agrevo Gmbh Synergistische herbizide Mischungen
US6037454A (en) 1996-11-27 2000-03-14 Genentech, Inc. Humanized anti-CD11a antibodies
KR100532178B1 (ko) * 1996-11-27 2005-12-01 제넨테크, 인크. 인간화 항-ＣＤ11ａ 항체
DE69738539T2 (de) 1996-12-03 2009-03-26 Amgen Fremont Inc. Vollkommen humane Antikörper die EGFR binden
US20050197285A1 (en) 1997-03-07 2005-09-08 Rosen Craig A. Human secreted proteins
US6040498A (en) 1998-08-11 2000-03-21 North Caroline State University Genetically engineered duckweed
KR20010023325A (ko) 1997-08-26 2001-03-26 리스 데브라 케이. 함지방세포-특이적 단백질 상동체
JP4460155B2 (ja) 1997-12-05 2010-05-12 ザ・スクリプス・リサーチ・インステイチユート マウス抗体のヒト化
GB9809951D0 (en) 1998-05-08 1998-07-08 Univ Cambridge Tech Binding molecules
US20030170781A1 (en) * 1999-07-01 2003-09-11 Zymogenetics, Inc. Secreted protein zacrp4
US7125978B1 (en) 1999-10-04 2006-10-24 Medicago Inc. Promoter for regulating expression of foreign genes
JP2003512821A (ja) 1999-10-04 2003-04-08 メディカゴ インコーポレイテッド 外来性遺伝子の転写調節方法
WO2002005634A2 (en) 2000-07-13 2002-01-24 University Of South Florida Transgenic animal and methods
US6872559B2 (en) 2000-10-26 2005-03-29 Wisconsin Alumni Research Foundation E. coli O157:H7 C1 esterase inhibitor-binding protein and methods of use
AU2002233340B2 (en) 2001-02-19 2008-05-22 Merck Patent Gmbh Artificial fusion proteins with reduced immunogenicity
CA2478313A1 (en) * 2001-11-07 2003-06-26 Human Genome Sciences, Inc. 41 human secreted proteins
US20040248156A1 (en) * 2001-12-03 2004-12-09 Tianhua Hu Methods and materials relating to novel C1q domain-containing polypeptides and polynucleotides
US20050214786A1 (en) 2002-02-22 2005-09-29 Human Genome Sciences, Inc. 26 human secreted proteins
JP2006504971A (ja) 2002-11-01 2006-02-09 ザ・リージェンツ・オブ・ザ・ユニバーシティ・オブ・コロラド，ア・ボディー・コーポレイト マトリックス支援レーザー脱離イオン化−飛行時間型質量分析によるタンパク質アイソフォームの定量的解析
JP4768620B2 (ja) 2003-05-15 2011-09-07 ジェネンテック， インコーポレイテッド 敗血症の予防及び治療のための方法及び組成物
WO2005002513A2 (en) * 2003-06-16 2005-01-13 The Brigham And Women's Hospital, Inc. C1q complement inhibitors and methods of use thereof
WO2005002512A2 (en) 2003-06-19 2005-01-13 University Of Rochester Immunotherapeutic vaccine strategy
US8501705B2 (en) * 2003-09-11 2013-08-06 The Board Of Regents Of The University Of Texas System Methods and materials for treating autoimmune and/or complement mediated diseases and conditions
US7544855B2 (en) 2004-04-23 2009-06-09 Buck Institute Transgenic mouse whose genome comprises an APP having a mutation at amino acid 664
US20080008719A1 (en) 2004-07-10 2008-01-10 Bowdish Katherine S Methods and compositions for the treatment of prostate cancer
US8420783B2 (en) 2004-12-08 2013-04-16 Immunomedics, Inc. Method and compositions for immunotherapy of inflammatory and immune-dysregulatory diseases
KR20070107687A (ko) 2004-12-31 2007-11-07 제넨테크, 인크. Ｂｒ３과 결합하는 폴리펩티드, 및 그의 용도
AU2005227420B2 (en) * 2005-10-14 2013-05-09 Alltech, Inc. Method and compositions for altering cell function
US8148330B2 (en) 2005-12-09 2012-04-03 The Board Of Trustees Of The Leland Stanford Junior University Modulation of synaptic maintenance
US9480658B2 (en) 2005-12-09 2016-11-01 The Board Of Trustees Of The Leland Stanford Junior University Modulation of synaptic maintenance
BRPI0709050B1 (pt) * 2006-03-23 2018-12-26 Bioarctic Neuroscience Ab anticorpo ou fragmento do mesmo, composição, métodos para detectar protofibrilas ab in vitro, e, uso do anticorpo
US7919079B2 (en) 2006-03-31 2011-04-05 Biosante Pharmaceuticals, Inc. Cancer immunotherapy compositions and methods of use
EP2380907B1 (en) 2006-09-05 2016-11-30 Alexion Pharmaceuticals, Inc. Methods and compositions for the treatment of antibody mediated neuropathies
CN101657097A (zh) 2007-03-01 2010-02-24 先进视觉疗法公司 以炎症为特征的疾病的治疗
US8877688B2 (en) 2007-09-14 2014-11-04 Adimab, Llc Rationally designed, synthetic antibody libraries and uses therefor
CA2697193C (en) 2007-09-14 2017-06-06 Adimab, Inc. Rationally designed, synthetic antibody libraries and uses therefor
MX2010006422A (es) 2007-12-11 2010-06-25 Glaxo Group Ltd Proteinas de union a antigenos.
CN105866426B (zh) 2008-12-01 2018-11-23 小利兰·斯坦福大学托管委员会 用于检测补体结合性抗体的方法和组合物
EP2426916A4 (en) 2009-04-28 2017-09-06 Panasonic Corporation Receiver
WO2012009568A2 (en) 2010-07-16 2012-01-19 Adimab, Llc Antibody libraries
WO2012067267A1 (en) 2010-11-18 2012-05-24 Takahiro Ochi TESTING METHOD USING ANTI-C1q MONOCLONAL ANTIBODY
PH12013502421A1 (en) 2011-05-27 2014-01-06 Glaxo Group Ltd Bcma (cd269/tnfrsf17) -binding proteins
US9127061B2 (en) 2011-06-24 2015-09-08 Perseus Proteomics Inc. Anti-human P-cadherin (CDH3) recombinant antibody
PL2794905T3 (pl) 2011-12-20 2020-11-02 Medimmune, Llc Zmodyfikowane polipeptydy dla rusztowań przeciwciał dwuswoistych
CN118924900A (zh) 2012-06-18 2024-11-12 奥默罗斯公司 抑制masp-1和/或masp-2和/或masp-3的组合物和方法
RS63212B1 (sr) 2012-11-02 2022-06-30 Bioverativ Usa Inc Antikomplementna c1s antitela i njihove primene
WO2014161570A1 (en) 2013-04-03 2014-10-09 Roche Glycart Ag Antibodies against human il17 and uses thereof
US20160053023A1 (en) 2013-04-09 2016-02-25 Annexon, Inc. Methods of treatment for neuromyelitis optica
EP2996722A4 (en) 2013-05-15 2017-01-11 Annexon, Inc. Methods of treatment for guillain-barre syndrome
KR102321320B1 (ko) 2013-07-09 2021-11-03 애넥슨, 인코포레이티드 항-보체 인자 ｃ１ｑ 항체 및 이의 용도
US10316081B2 (en) 2014-11-05 2019-06-11 Annexon, Inc. Humanized anti-complement factor C1Q antibodies
BR112018010360A2 (en) 2015-11-24 2018-12-04 Annexon, Inc. anti-c1q complement factor fab fragments and uses thereof
US20190161535A1 (en) 2016-05-09 2019-05-30 Annexon, Inc. Compositions and methods for treating spinal muscular atrophy
WO2018017711A1 (en) 2016-07-19 2018-01-25 Annexon, Inc. Compositions and methods for treating frontotemporal dementia
CA3200976A1 (en) 2020-12-04 2022-06-09 Anita GROVER Compositions and methods for treating ocular diseases

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150259437A1 (en) * 2012-10-25 2015-09-17 True North Therapeutics, Inc. Anti-complement c1s antibodies and uses thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Hampel et al. The future of Alzheimer's disease: the next 10 years. Prog Neurobiol. 2011 Dec;95(4):718-28. Epub 2011 Nov 22. *
Perrin et al. Multimodal techniques for diagnosis and prognosis of Alzheimer's disease. Nature 2009 Oct 15;461(7266):916-22, Epub 2009 Oct 14. *
Vickers. A vaccine against Alzheimer's disease: developments to date. Drugs Aging 2002; 19(7):487-94. *

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12215169B2 (en) 2012-10-25 2025-02-04 Bioverativ Usa Inc. Anti-complement C1s antibodies and uses thereof
US10457745B2 (en) 2012-10-25 2019-10-29 Bioverativ Usa Inc. Anti-complement C1s antibodies
US12240917B2 (en) 2012-11-02 2025-03-04 Bioverativ Usa Inc. Anti-complement C1s antibodies and uses thereof
US10450382B2 (en) 2012-11-02 2019-10-22 Bioverativ Usa Inc. Anti-complement C1s antibodies
US10227398B2 (en) 2013-07-09 2019-03-12 Annexon, Inc. Anti-complement factor C1q antibodies and uses thereof
US10590190B2 (en) 2013-07-09 2020-03-17 Annexon, Inc. Anti-complement factor C1q antibodies and uses thereof
US10927167B2 (en) 2013-07-09 2021-02-23 Annexon, Inc. Anti-complement factor C1Q antibodies and uses thereof
US11649279B2 (en) 2013-07-09 2023-05-16 Annexon, Inc. Anti-complement factor C1Q antibodies and uses thereof
US10316081B2 (en) * 2014-11-05 2019-06-11 Annexon, Inc. Humanized anti-complement factor C1Q antibodies
US20160355574A1 (en) * 2014-11-05 2016-12-08 Arnon Rosenthal Humanized anti-complement factor c1q antibodies and uses thereof
US11246926B2 (en) 2015-04-06 2022-02-15 Bioverativ Usa Inc. Polynucleotides encoding anti-C1s antibodies
US10729767B2 (en) 2015-04-06 2020-08-04 Bioverativ Usa Inc. Humanized anti-C1s antibodies and methods of inhibiting C1s cleavage
US10723788B2 (en) 2015-11-24 2020-07-28 Annexon, Inc. Anti-complement factor C1q Fab fragments and uses thereof
US11999779B2 (en) 2015-11-24 2024-06-04 Annexon, Inc. Anti-complement factor C1q Fab fragments and uses thereof
US12391750B2 (en) 2016-10-12 2025-08-19 Bioverativ Usa Inc. Anti-C1S antibodies and methods of use thereof
KR102638884B1 (ko) 2016-10-12 2024-02-22 바이오버라티브 유에스에이 인코포레이티드 항-C1s 항체 및 그의 사용 방법
KR20190082783A (ko) * 2016-10-12 2019-07-10 바이오버라티브 유에스에이 인코포레이티드 항-C1s 항체 및 그의 사용 방법
WO2018071676A1 (en) * 2016-10-12 2018-04-19 Bioverativ Usa Inc. Anti-c1s antibodies and methods of use thereof
KR102891406B1 (ko) 2016-10-12 2025-11-27 바이오버라티브 유에스에이 인코포레이티드 항-C1s 항체 및 그의 사용 방법
EP4045084A4 (en) * 2019-10-17 2023-05-31 Annexon, Inc. COMPOSITIONS AND METHODS FOR TREATING BLOOD DISORDERS
WO2021226513A1 (en) 2020-05-08 2021-11-11 President And Fellows Of Harvard College Methods for treating inflammatory and autoimmune disorders
US12486322B2 (en) 2021-12-13 2025-12-02 Annexon, Inc. Anti-complement factor C1q antibodies with single binding arms and uses thereof
WO2023130001A1 (en) * 2021-12-30 2023-07-06 Annexon, Inc. Compositions and methods for treating huntington's disease
CN120685919A (zh) * 2025-08-22 2025-09-23 中国科学院长春应用化学研究所 一种C1q补体蛋白在制备检测Aβ42蛋白的元件中的应用、一种检测Aβ42蛋白的芯片及其应用

Also Published As

Publication number Publication date
NZ715455A (en) 2021-12-24
EP3019240A4 (en) 2017-01-04
CA2916521A1 (en) 2015-01-15
PL3019240T3 (pl) 2024-08-19
CA2916521C (en) 2023-03-07
PT3019240T (pt) 2024-06-06
AU2014287221C1 (en) 2020-03-05
ES2978917T3 (es) 2024-09-23
FI3019240T3 (fi) 2024-05-30
EP3019523A4 (en) 2016-12-28
DK3019240T3 (da) 2024-06-03
IL243506B (en) 2021-07-29
US20190211084A1 (en) 2019-07-11
SG10202008809RA (en) 2020-10-29
AU2014287221B2 (en) 2019-09-12
WO2015006507A1 (en) 2015-01-15
US10590190B2 (en) 2020-03-17
SI3019240T1 (sl) 2024-07-31
SG11201510316UA (en) 2016-01-28
US10927167B2 (en) 2021-02-23
US20170334976A1 (en) 2017-11-23
US9708394B2 (en) 2017-07-18
US20210277096A1 (en) 2021-09-09
JP2019071889A (ja) 2019-05-16
JP6653009B2 (ja) 2020-02-26
US10227398B2 (en) 2019-03-12
MY175896A (en) 2020-07-14
US20160368973A1 (en) 2016-12-22
US11649279B2 (en) 2023-05-16
KR102321320B1 (ko) 2021-11-03
EA037325B1 (ru) 2021-03-12
JP2016526393A (ja) 2016-09-05
MX2020010141A (es) 2020-10-20
IL243506A0 (en) 2016-02-29
EA201690182A1 (ru) 2016-10-31
ZA201600157B (en) 2025-10-29
EP3019240A1 (en) 2016-05-18
CN105682740A (zh) 2016-06-15
US20200239557A1 (en) 2020-07-30
KR20160058752A (ko) 2016-05-25
WO2015006504A1 (en) 2015-01-15
EP3019240B1 (en) 2024-03-13
EP3019523A1 (en) 2016-05-18
MX375500B (es) 2025-03-06
BR112016000106A2 (pt) 2017-12-12
JP6462680B2 (ja) 2019-01-30
HUE066722T2 (hu) 2024-09-28
EP4252769A2 (en) 2023-10-04
BR112016000106B1 (pt) 2023-11-21
EP4252769A3 (en) 2023-11-29
CN105682740B (zh) 2021-06-11
US20240083987A1 (en) 2024-03-14
SG10201710758PA (en) 2018-02-27
MX2015017341A (es) 2016-07-06
AU2014287221A1 (en) 2016-01-21
HK1225678A1 (zh) 2017-09-15

Similar Documents

Publication Publication Date Title
US20240083987A1 (en) Anti-complement factor c1q antibodies and uses thereof
US20160090425A1 (en) Anti-complement factor c1s antibodies and uses thereof
EP3280441B1 (en) Anti-sortilin antibodies and methods of use thereof
EP2983710B1 (en) Methods of treatment for neuromyelitis optica
Class et al. Patent application title: ANTI-COMPLEMENT FACTOR C1S ANTIBODIES AND USES THEREOF Inventors: Arnon Rosenthal (Woodside, CA, US) Michael Leviten (Palo Alto, CA, US)
HK1225678B (zh) 抗-补体因子c1q抗体及其应用

Legal Events

Date Code Title Description
AS Assignment

Owner name: CHILDREN'S MEDICAL CENTER CORPORATION, MASSACHUSET

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:STEVENS, BETH A.;HONG, SOYON;WILTON, DANIEL;REEL/FRAME:037839/0220

Effective date: 20160219

Owner name: ANNEXON, INC., CALIFORNIA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ROSENTHAL, ARNON;LEVITEN, MICHAEL;REEL/FRAME:037839/0234

Effective date: 20160202

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION