+

WO2000021559A2 - Facteur de blocage b destine a traiter une maladie d'origine immunitaire induite par complement - Google Patents

Facteur de blocage b destine a traiter une maladie d'origine immunitaire induite par complement Download PDF

Info

Publication number
WO2000021559A2
WO2000021559A2 PCT/US1999/023485 US9923485W WO0021559A2 WO 2000021559 A2 WO2000021559 A2 WO 2000021559A2 US 9923485 W US9923485 W US 9923485W WO 0021559 A2 WO0021559 A2 WO 0021559A2
Authority
WO
WIPO (PCT)
Prior art keywords
factor
subject
mice
antibody
activity
Prior art date
Application number
PCT/US1999/023485
Other languages
English (en)
Other versions
WO2000021559A3 (fr
Inventor
Gary Gilkeson
Hiroshi Watanabe
Original Assignee
Musc Foundation For Research Development
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 Musc Foundation For Research Development filed Critical Musc Foundation For Research Development
Priority to AU17051/00A priority Critical patent/AU1705100A/en
Publication of WO2000021559A2 publication Critical patent/WO2000021559A2/fr
Publication of WO2000021559A3 publication Critical patent/WO2000021559A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A01K67/0275—Genetically modified vertebrates, e.g. transgenic
    • A01K67/0276—Knock-out vertebrates
    • 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
    • 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
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/647—Blood coagulation factors not provided for in a preceding group or according to more than one of the proceeding groups
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/07—Animals genetically altered by homologous recombination
    • A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00—Animals characterised by species
    • A01K2227/10—Mammal
    • A01K2227/105—Murine
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/0306—Animal model for genetic diseases
    • A01K2267/0325—Animal model for autoimmune diseases
    • 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
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy

Definitions

  • the present invention relates to the field of complement-mediated autoimmune diseases.
  • the present invention relates to the role of the alternative pathway in complement activation and specifically blocking Factor B to treat and prevent complement-mediated autoimmune diseases.
  • C3bBb complex is a C3 convertase and leads to the formation of more molecules of C3b resulting in activation of additional Bf setting up an amplification loop for activation of C3 (1-3).
  • C3 activation also initiates the cascade of events that leads to immune complex formation.
  • C5 convertase C3bBbC3b follows with activation of terminal complement components on the pathogen surface. This process leads to opsonization and direct damage of the pathogen (1-3).
  • Factor B is encoded by a single gene in the MHC Class III gene cluster immediately 3' to the C2 gene; C2 is the functional homologue of Bf in the classical complement pathway (4, 5).
  • Factor B is produced by hepatocytes (6), phagocytes (7, 8), fibroblasts (9), epithelial cells (10), and endothelial cells (11).
  • Bf is an acute phase reactant; therefore, serum levels of Bf increase during inflammation stimulated by cytokines, growth factors, and bacterial products (1, 5).
  • Cis-elements responsible for the constitutive and regulated expression of Bf are located in the C2-Bf intergenic region (4).
  • Factor B is not only a component of the alternative complement pathway, but also serves as a B cell growth factor (12, 13), stimulates mononuclear cell cytotoxicity (14, 15), induces macrophage spreading (16), and solubilizes immune complexes (ICs) (17-19).
  • ICs immune complexes
  • the classical pathway of the complement system has a known important role in immune complex glomerulonephritis. ICs depositing in the kidney activate the classical pathway, thus eliciting inflammation and tissue destruction (1, 5, 22). Furthermore, components of the classical complement pathway are important in humoral responses to foreign antigens, immune complex clearance, B cell tolerance, and disposition of apoptotic cells (23-27). Genetic deficiency of C3 and therapies directed at blocking C3 activity (e.g. Crry transgenic mice) prevent glomerular inflammation and tissue damage in murine models of induced glomerulonephritis (28- 32).
  • the present invention overcomes previous limitations in the art by providing methods of blocking Factor B, antibodies directed against Factor B, peptides which block the receptors of Factor B and an oligonucleotide which inhibits synthesis of Factor B.
  • the present invention provides a method of treating or preventing Factor B- mediated immune disease in a subject, comprising administering to the subject an inhibitor of synthesis or activity of Factor B, whereby the inhibition of synthesis or activity of Factor B treats or prevents the immune disease.
  • the present invention further provides an anti-sense oligonucleotide capable of hybridizing to the nucleic acid which encodes the protein, Factor B.
  • the present invention also provides a peptide that mimics the binding properties of Factor B but lacks the ability to activate C3, whereby the binding of the peptide to C3 does not activate C3 and blocks the effects of Factor B on C3, thereby reducing formation and deposition of immune complexes in the subject.
  • the present invention provides a method of determining the predisposition of a subject to a complement-mediated disease, comprising measuring the serum level and activity of Factor B, and correlating the amount of complement activation with the serum level of factor B.
  • a method of diagnosing a subject as having a Factor B- mediated immune disease comprising measuring the serum level and activity of Factor B and determining the amount of complement activation.
  • the present invention also provides a method of determining the efficacy of treatment in and prognosis of a subject with a Factor B-mediated immune disease, comprising measuring the serum level and activity of Factor B and determining the amount of complement activation.
  • transgenic mouse comprising a mutation in the Fas gene and a mutation in the gene producing Factor B.
  • the present invention provides a method of determining the efficacy of treatment in and prognosis of a subject with Factor B-mediated immune disease, comprising measuring the serum level and activity of C3, whereby a lower than normal serum level of C3 indicates a poor prognosis and lack of efficacy of treatment, and whereby a rising serum level of C3 from a lower than normal level to a nearly normal level of C3 indicates efficacy of treatment and an improving prognosis.
  • FIG. 1 Urinary protein excretion by MRL/lpr B-/-, B+/+, and B+/- mice. Data presented are the mean 24 hour protein excretion (mg/mouse/day) of 6 mice +/- standard error in each group. p ⁇ 0.01 B+/- mice versus B-/- mice; p ⁇ 0.05 B+/+ mice versus B-/- mice; p ⁇ 0.05 B+/- mice versus B+/+ mice.
  • FIG. 1 Renal pathology of kidneys from MRL/lpr B-/-, B+/+, and B+/- mice.
  • A) Data presented are the mean renal score +/- standard error in each group. There were no statistically significant differences between B-/- and other groups at the age of 22 weeks (p 0.13). At the age of 44 weeks, there was a statistically significant difference between B-/- and B+/+ mice (p- .04).
  • FIG. 1 IgG3 cryoglobuhn levels in serum of MRL/lpr mice. Data shown are the mean serum IgG3 cryoglobuhn levels ( ⁇ g/ml) +/- standard error in each group. Cryoglobuhn IgG3 levels in serum of B-/- mice were significantly decreased at each time point. p ⁇ 0.01 versus B+/+ mice. The level of IgG3 in the serum of B+/- mice was decreased due to the death of mice with severe disease by 24 weeks of age.
  • a or “an” or “the” may mean one or more.
  • a cell may mean one cell or more than one cell.
  • the present invention is based, at least in part, on the unexpected and surprising discovery that blocking the synthesis or activity of Factor B significantly reduced the incidence of Factor B-mediated immune disease in a subject.
  • the present invention provides a new means for preventing and treating Factor B-mediated autoimmune disease in mammals, especially human beings.
  • the present invention provides a method of treating or preventing complement- mediated immune disease in a subject, comprising administering to the subject an inhibitor of synthesis or activity of Factor B, whereby the inhibition of synthesis or activity of Factor B treats or prevents the immune disease.
  • the immune disease treated in the present invention is mediated at least in part by the activation of Factor B in the alternative complement pathway.
  • complement-mediated immune diseases There are many known complement-mediated immune diseases.
  • Factor B may be instrumental in activating the following diseases: vasculitis, systemic lupus erythematosus, rheumatoid arthritis, myocardial infarction, ischemic/ reperfusion injury, cerebrovascular accident, Alzheimer's disease, transplantation rejection (xeno and allo), all antibody-mediated skin diseases, all antibody-mediated organ-specific diseases (including Type I and Type II diabetes mellitus, thyroiditis, idiopathic thrombocytopenic purpura and hemolytic anemia, and neuropathies), multiple sclerosis, cardiopulmonary bypass injury, membranoproliferative glomerulonephritis, polyarteritis nodosa, Henoch Schonlein purpura, serum sickness, Goodpasture's disease, systemic necrotizing vasculitis, post streptococcal glomerulonephritis, idiopathic pulmonary fibrosis (usual interstitial pneumonitis) and membranous glomerulonep
  • the present invention provides a method for contacting the subject with an antibody to Factor B, under conditions that permit the binding of the antibody to Factor B in the subject.
  • This binding of the antibody to Factor B reduces the serum level or activity of Factor B, thereby reducing the amount of Factor B available to begin the cascade of events in the alternative complement pathway which enhances activation of the classical complement pathway.
  • With less activation of the classical complement pathway less inflammation occurs.
  • fewer immune complexes are formed which can be deposited in the organs, thereby decreasing the incidence of disease.
  • the present invention also provides a method wherein the binding of antibody to Factor B reduces the activation of C3, thereby decreasing complement-induced inflammation.
  • C3 is an enzyme in the complement cascade on which both the classical and alternative pathways act.
  • This antibody may be monoclonal or polyclonal and may be produced by one of ordinary skill in the art.
  • a purified monoclonal antibody that specifically binds and blocks Factor B is provided.
  • a purified polyclonal antibody that specifically binds and blocks Factor B is also provided.
  • the antibody can specifically bind a unique epitope of Factor B.
  • bind means the well understood antigen antibody binding as well as other nonrandom association with an antigen.
  • Specifically bind as used herein describes an antibody or other ligand that does not cross react substantially with any antigen other than the one specified, in this case, Factor B.
  • Blocks as used herein means interferes with or prevents the action of.”
  • Factor B or an immunogenic fragment thereof can be injected into an animal in an amount and in intervals sufficient to elicit an immune response.
  • Polyclonal antibodies can be purified directly, or spleen cells from the animal can be fused with an immortal cell line and screened for monoclonal antibody secretion.
  • purified monospecific polyclonal antibodies that specifically bind the antigen are within the scope of the present invention.
  • the present invention further provides a composition comprising an antibody to Factor B in a pharmaceutically acceptable carrier.
  • the antibody can be administered to a subject by parenteral administration.
  • parenteral administration includes intradermal, subcutaneous, intramuscular, intraperitoneal, intravenous and intratracheal routes.
  • parenteral administration includes intradermal, subcutaneous, intramuscular, intraperitoneal, intravenous and intratracheal routes.
  • a more recently revised approach for parenteral administration involves use of a slow release or sustained release system such that a constant dosage is maintained. See, e.g., U.S. Patent No. 3,610,795, which is incorporated by reference herein.
  • To determine the dosage of the composition to be used in treating Factor B- mediated immune disease a person of skill in the art would infuse the antibody into a subject after first determining the level of Factor B in the serum of the subject. The dosage in mg/Kg would be based on that amount of antibody necessary to eliminate Factor B from the serum and the activity of
  • the efficacy of treatment with an anti-Factor B antibody is confirmed in a murine model of Factor B-mediated glomerulonephritis, that is accepted in this field as representative of human glomerulonephritis.
  • the present invention also provides a method of blocking Factor B comprising administering an anti-sense oligonucleotide to block the synthesis of Factor B in a cell of the subject in vivo.
  • the anti-sense oligonucleotide interferes with the mRNA necessary for the synthesis of Factor B in the cell and reduces the serum level of Factor B, thereby reducing formation and deposition of immune complexes in the subject. Moreover, the reduction of the serum level of Factor B decreases the level of activation of C3 in the classical complement pathway, thereby reducing the incidence of complement-mediated inflammation and immune disease.
  • the present invention provides an anti-sense oligonucleotide capable of hybridizing to an mRNA encoded by the Factor B coding sequence (49).
  • Antisense technology is well known in the art and describes a mechanism whereby a nucleic acid comprising a nucleotide sequence which is in a complementary, "antisense” orientation with respect to a coding or “sense” sequence of an endogenous gene is introduced into a cell, whereby a duplex forms between the antisense sequence and its complementary sense sequence. The formation of this duplex results in inactivation of the endogenous gene.
  • Antisense nucleic acids can be produced for any endogenous gene for which the coding sequence has been or can be determined according to well known methods.
  • Antisense nucleic acids can inhibit gene expression by forming an RNA/RNA duplex between the antisense RNA and the RNA transcribed from a target gene.
  • the precise mechanism by which this duplex formation decreases the production of the protein encoded by the endogenous gene most likely involves binding of complementary regions of the normal sense mRNA and the antisense RNA strand with duplex formation in a manner that blocks RNA processing and translation.
  • Alternative mechanisms include the formation of a triplex between the antisense RNA and duplex DNA or the formation of a DNA-RNA duplex with subsequent degradation of DNA-RNA hybrids by RNAse H.
  • an antisense effect can result from certain DNA-based oligonucleotides via triple-helix formation between the oligomer and double-stranded DNA which results in the repression of gene transcription.
  • An expression construct including a vector may be used to introduce a nucleic acid encoding the anti-sense oligonucleotide into a cell of the subject.
  • the exact method of introducing the nucleic acid into mammalian cells is, of course, not limited to the use of adenoviral vectors.
  • Other vector systems can be utilized in this invention including the retroviral vector system which can package a recombinant retroviral genome. The recombinant retrovirus can then be used to infect and thereby deliver nucleic acid to the infected cells.
  • AAV adeno-associated viral
  • HSV herpes virus
  • lentiviral vectors lentiviral vectors
  • pseudotyped retroviral vectors lentiviral vectors
  • pox virus vectors such as vaccinia virus vectors.
  • Physical transduction techniques can also be used, such as liposome delivery and receptor-mediated and other endocytosis mechanism.
  • This invention can be used in conjunction with any of these or other commonly used gene transfer methods.
  • the present invention further provides a vector in a composition comprising a pharmaceutically acceptable carrier.
  • Delivery of the nucleic acid or vector to cells can be via a variety of mechanisms.
  • delivery can be via a liposome, using commercially available liposome preparations such as LIPOFECTIN, LIPOFECTAMINE (GIBCO-BRL, Inc., Gaithersburg, MD), SUPERFECT (Qiagen, Inc. Hilden, Germany) and TRANSFECTAM (Promega Biotec, Inc., Madison, WI), as well as other liposomes developed according to procedures standard in the art.
  • the nucleic acid or vector of this invention can be delivered in vivo by electroporation, the technology for which is available from Genetronics, Inc. (San Diego, CA) as well as by means of a SONOPORATION machine (ImaRx Pharmaceutical Corp., Arlington, AZ).
  • the present invention provides a peptide that mimics the binding properties of Factor B but lacks the ability to activate C3, whereby the binding of the peptide to C3 does not activate C3 and blocks the effects of Factor B on C3, thereby reducing formation and deposition of immune complexes in the subject.
  • the peptide of the present invention may bind the same region of C3 to which Factor B normally binds or any other region of C3, whereby the binding of the peptide to C3 blocks the binding of Factor B to C3. Therefore, the peptide can comprise the sequence of amino acids of the known Factor B sequence that is known to interact with or bind C3.
  • a peptide may bind to Factor B in that region of Factor B which binds to C3, thereby blocking Factor B from binding to and activating C3.
  • This sequence can correspond to the region of amino acids of C3 that interact or bind to Factor B.
  • the present invention provides a method for blocking the effect of Factor B on C3, comprising administering to a subject a peptide that mimics the binding properties of Factor B but lacks the ability to activate C3.
  • the binding of the peptide to C3 does not activate C3 and competitively blocks the effects of Factor B on C3, thereby reducing formation and deposition of immune complexes in the subject.
  • the peptide binding to C3 blocks activation of C3, thereby decreasing complement-mediated inflammation and immune disease.
  • the present invention provides a composition comprising a peptide which binds C3 and a pharmaceutically acceptable carrier.
  • the composition can be administered parenterally, including but not limited to intravenously, intramuscularly and intraperitoneally.
  • the present invention provides an isolated nucleic acid encoding the peptide which binds but does not activate C3.
  • a vector comprising the nucleic acid encoding the peptide is provided.
  • the vector can be a commercially available preparation or can be constructed in the laboratory according to methods well known in the art. This vector may be in a cell in vivo or ex vivo.
  • the present invention also provides a composition comprising the vector and a pharmaceutically acceptable carrier.
  • the present invention provides a method of determining the predisposition of a subject to Factor B-mediated immune disease, comprising measuring the serum level and activity of Factor B and determining the amount of complement activation. A higher than normal level of Factor B indicates complement-mediated immune disease and active inflammation.
  • the present invention also provides a method of diagnosing a subject as having Factor B-mediated immune disease, comprising measuring the level of anti-DNA antibody in the serum of the subject. A higher than normal level of anti-DNA antibody indicates complement-mediated immune disease and active inflammation.
  • Anti-DNA antibody can be measured in the serum according to the methods taught in the Examples.
  • Factor B can be measured in the serum by the method taught in the Examples. Serum levels of Factor B can be measured in a population of subjects in whom there is no sign of complement-mediated immune disease to determine a normal range of values. With the same method, a person skilled in the art can measure the serum level of Factor B in a subject with signs of complement-mediated immune disease to determine an abnormal range of values. A higher than normal level of Factor B in a subject would indicate the presence of inflammation and complement-mediated immune disease. Likewise, a cohort can be followed over the long term to identify sub ranges measured within the apparently normal population that predict progression to a diseased state.
  • the present invention also provides a method of determining the efficacy of treatment in and prognosis of a subject with complement-mediated immune disease, comprising measuring the serum level and activity of Factor B and determining the amount of complement activation.
  • the clinician would expect a higher than normal serum level of Factor B to fall to a nearly normal level, if the treatment is effective. Absence of a fall in the serum level of Factor B would indicate lack of efficacy of a therapeutic regimen and continued activation of complement with continuing deposition of immune complexes.
  • the present invention also provides a transgenic mouse comprising a mutation in the Fas gene and a mutation in the gene producing Factor B.
  • This mutant mouse is the MRL-lpr Factor B knockout mouse.
  • the present invention provides a method of determining the efficacy of treatment in and prognosis of a subject with Factor B-activated complement-mediated immune disease, comprising measuring the serum level and activity of C3, whereby a lower than normal serum level of C3 indicates a poor prognosis and lack of efficacy of treatment, and whereby a rising serum level of C3 from a lower than normal level to a nearly normal level of C3 indicates efficacy of treatment and an improving prognosis.
  • the method of measuring the serum level of C3 is taught in the Examples.
  • C5A and membrane attack complexes can be measured to monitor the efficacy of treatment in accordance with methods known to a person of skill in the art.
  • Factor B deficient mice Factor B deficient mice were derived as previously described (36). B6/129 B+/- mice were intercrossed for 4 generations to yield the B-/- mice used in these studies. A B6/129 intercross B-/- female was bred to an MRL/lpr male. Offspring of this mating were further backcrossed for 3 generations to MRL/lpr mice. B+/- backcross 4 mice were bred to yield the mice used in this study. Eleven B-/-, fourteen B+/+ and ten B+/- mice were included in this study. Mice were maintained in the animal facility of Washington University and the Strom Thurmond Biomedical
  • PCR was performed by adding 2ml of DNA (l ⁇ g) into a 23 ⁇ l reaction mixture containing 1.5mM of MgC12, 6.7 ⁇ M each of oligonucleotide mix, lOmM each of dNTP mix, and 0.2ml of Taq Gold.
  • PCR conditions were as follows: 94°C for 4 min.; 28 cycles of 94°C for 1 min., 68 ° C for 1 min., and 72°C for 2 min; 72°C for 10 min.
  • samples were electrophoresed in a 2% agarose gel and visualized by ethidium bromide staining.
  • Fas A (5'-AGGTTAC
  • H2- H2 of the mice was determined by flow cytometric analysis of splenic mononuclear cells. Single cell suspensions were derived and stained with FITC labeled anti-H2D k (monoclonal antibody KH95) or anti-H2D b (monoclonal antibody 15-5-5, Pharmingen, San Diego, CA). FITC labeled isotype control antibodies were used to eliminate non-specific staining as a variable. Samples were analyzed on a Becton Dickinson FACSTAR machine and results analyzed using CellQuest software (Becton Dickinson).
  • PCR RFLP analysis was performed using the techniques of Peng and Craft specifically designed to differentiate H2 k and H2 D (37). Briefly, tail DNA was amplified using the primers IAAIF 5 ' GAAGACGACAT TGAGGCCGACCACGTAGGC 3 ' (SEQ ID NO:7) and IAALR 5' ATTGGTAGCTGGGGTGGAATTTG ACCTCTT 3' (SEQ ID NO:8). PCR was performed with a 12 minute denaturation step of 94°C followed by cycles of 94°C for 20 sec, 60°C for 1 minute and 72°C for 1 minute with a final extension of 10 minutes at 72°C.
  • the resultant PCR product was cut with Hind III and run in an agarose gel. Amplified DNA of the H2 k genotype is cut while DNA of the H2 b genotype is not allowing differentiation on agarose gel electrophoresis.
  • Urine protein excretion Mice were placed in metabolic cages for 24 hour urine collection. To prevent bacterial growth, antibiotics (ampicillin, gentamicin, and chloramphenicol) were added to collection tubes. Urinary protein excretion was determined using the BioRad protein assay kit according to the manufacturer's instructions (BioRad, Hercules, CA) and reported as mg of protein/mouse/day.
  • Anti-DNA levels were measured by ELISA as previously described (38). Briefly, 96 well ELISA plates were coated with 5 ⁇ g/ml of double stranded calf thymus DNA (dsDNA) at 37°C overnight. The plates were then washed with PBS 0.05% tween (PBS-T). Sera were added in serial dilutions starting at a 1/100 dilution to each well, and incubated for 45 min at room temperature (RT). After washing with PBS-T, horse radish peroxidase (HRP) conjugated goat anti-mouse IgG ( ⁇ - chain specific, Sigma Chemical Co., St. Louis, MO) was added and incubated for 45 min.
  • HRP horse radish peroxidase
  • dsDNA was derived by SI nuclease (Sigma, St. Louis, MO) treatment.
  • cryoglobulins were isolated from sera as previously described (39). Briefly, blood samples were placed at 37°C for 2 hours. After centrifugation at 1000 rpm, supernatants were collected and placed at 4°C for 72 hours. They were then centrifuged at 3000 rpm, supernatants removed, and precipitates washed 5 times with cold PBS. After washing, they were resuspended in PBS at the same volume as the original sera. The isolated cryoglobulins were placed at 37°C for 3 hours before use.
  • Total Ig Total IgG levels in sera or in cryoglobulins were determined by ELISA using a standard curve of known concentrations of mouse IgG. Cryoglobulins were placed at 37°C for 3 hours before use. ELISA plates were coated with l ⁇ g/ml anti- mouse K overnight at 4°C. Sera or cryoglobulins were added in serial dilutions starting at a 1/1000 dilution. HRP conjugated goat anti-mouse IgG ( ⁇ -specific, Sigma, St. Louis, MO) was added, followed by TMB for color development. OD380 absorbance was measured as above.
  • Circulating immune complexes were determined by the Clq ELISA method as previously described with minor modifications (40). ELISA plates were coated with human Clq in 0.1M Carbonate buffer (pH 9.6) and incubated for 48 hours at 4°C. After washing with PBS, PBS-1% BSA was added to each well and incubated for 2 hours at RT. EDTA treated sera samples (diluted 1/50) were added in duplicate, then incubated for lh at RT and then overnight at 4°C. HRP anti-mouse IgG ( ⁇ chain specific, Sigma) was added in PBS-T. The rest of the assay was performed as described above. Aggregated human ⁇ globulin was used as a positive control.
  • Rheumatoid factor Rheumatoid factor (RF) levels in sera were measured by ELISA as previously described (33). ELISA plates were coated with rabbit IgG overnight at 4°C. After washing, sera were added in serial dilution starting at 1/100 dilution. The assay was then performed as described above using HRP conjugated goat anti-mouse IgG ( ⁇ chain specific, Sigma) or goat anti-mouse IgM (Sigma). For the measurement of IgG3 anti-IgG2a RF, the same method was used except ELISA plates were coated with mouse IgG2a and HRP anti-mouse IgG3 was used for detection of RF activity.
  • C3 levels in sera were measured by ELISA.
  • 96 well ELISA plates were coated with goat anti-mouse C3 (Cappel, Durham, NC) and incubated overnight at 4°C.
  • samples diluted 1/200, 1/1000, 1/5000 and 1/25000 were added to individual wells, and incubated for l-2h at RT.
  • rat anti-mouse C3 monoclonal antibody RmCl 1H9 was added and incubated for lh at RT (28).
  • Peroxidase conjugated anti-rat IgG was added, then incubated for lh at RT.
  • ImM ATBS in 0.1M citric acid buffer and 0.03% H2O2 was added for color development. The plates were read on an ELISA reader at OD 405.
  • Factor B levels were measured by ELISA. ELISA plates were coated with goat anti-human Factor B IgG fraction (DiaSorin, Stillwater, MN) overnight at 4°C. Sera diluted to 1/100, l/200,and 1/400 were added to individual wells. Biotinylated anti-human Bf antibody (Diasorin) was added followed by avidin-peroxidase. TMB was added for color development. After incubation for 30 min, OD 380 absorption was determined.
  • kidneys were removed.
  • One kidney was fixed with formaldehyde, embedded in paraffin, then sectioned before staining with hematoxylin and eosin (H&E) and PAS.
  • the slides were read and interpreted in a blinded fashion grading the kidneys for glomerular inflammation, proliferation, crescent formation, and necrosis. Interstitial changes and vasculitis were also noted. Scores from 0-3 were assigned for each of these features and then added together to yield a final renal score (33). For example, glomerular inflammation was graded 0-normal, 1-few inflammatory cells, 2-moderate inflammation, 3-severe inflammation.
  • the other kidney was snap frozen in liquid nitrogen and placed in OCT medium.
  • H2 To assess the MHC expression by each group, H2 expression of splenocytes was assessed by FITC staining and flow cytometry analysis.
  • the MRL/lpr Bf deficient mice were H2D b .
  • the wildtype mice were H2D k .
  • PCR RFLP analysis of MHC Class II expression confirmed the H2 haplotvpe of the mice with the B- /- mice being H2 b .
  • B+/- mice were H2 b/k and the B+/+ mice were H2 k .
  • Bf levels in sera were measured by ELISA when the mice were
  • Urinary protein excretion To assess the role of Bf in autoimmune nephritis, 24 hour urine protein excretion was measured beginning at 12 weeks of age. As shown in Figure 1 , there was a statistically significant difference in proteinuria comparing B+/- mice after 20 weeks of age (p ⁇ 0.05) to both B+/+ mice and B-/- mice indicating the development of early severe disease in the heterozygous mice. The B+/+ mice developed increasing proteinuria after 26 weeks of age that was significantly greater than the B-/- mice (p ⁇ _0.05). Even at 44 weeks of age, B-/- mice still had not developed proteinuria >1 mg/mouse/day (data not shown).
  • Renal score A group of mice were sacrificed at the age of 22 weeks (5 B-/-, 7
  • B+/+, and 3 B+/- mice were determined using a previously published scale for glomerular inflammation (33).
  • the remaining B+/+ and B-/- mice were sacrificed at 44 weeks (all B+/- mice died prior to 44 weeks).
  • Bf deficient mice had lower renal scores than the mice in the other groups at both 22 and 44 weeks.
  • the renal abnormalities in the MRL/lpr B-/- mice consisted of focal glomerular hypercellularity without significant infiltrate of inflammatory cells (Figure 2B). Glomerular wire loop lesions, immune complex deposition, and glomerular hyalinization, as determined by intensity of PAS staining, were minimally present in the B-/- mice ( Figure 2C).
  • the difference in renal scores in the B-/- compared to the other 2 groups at 22 weeks was not statistically significant (p 0.13) due to severe disease in one male B-/- mouse whose renal score was similar to that seen in the other two groups.
  • Immunofluorescence analysis To determine the effects of Bf genotypes on glomerular IC deposition, immunofluorescence analysis was performed. Frozen sections were stained with fluorescein conjugated anti-mouse C3, IgG, IgG3, IgM, and IgA. C3, IgM, and IgA deposition were similar in all groups with no apparent qualitative or quantitative differences (data not shown). Glomerular IgG3 deposition was minimal in all three groups. There was, however, significantly less IgG deposition in B-/- compared to B+/+ mice (p ⁇ 0.05, Table 2). IgG deposition in the kidneys of B-/- mice was also less than in the B+/- mice; the difference, however, did not reach statistical significance (Table 2).
  • Vasculitis Medium to large vessel vasculitis is a pathologic feature of renal disease in MRL/lpr mice (41). At the age of 22 weeks, of the five B-/- mice examined, only the male B-/- mouse with severe renal disease had vasculitis (1/5). 50%o of female B+/+ mice (2/4) had vasculitis as illustrated in Figure 2F, while no vasculitis was present in male B+/+ mice (0/3). In the B+/- mice, 1/3 of the mice had vasculitis; all three B+/- mice studied were females. At the age of 44 weeks, 100% of female B+/+ mice had vasculitis (3/3).
  • vasculitis in male B+/+ mice There was no vasculitis in male B+/+ mice (0/4) nor vasculitis observed in the 6 B-/- mice (3 males and 3 females). These data indicate a greater incidence of vasculitis in the B+/+ mice than in the B-/- mice (overall 1/11 B-/- mice had vasculitis compared to 5/14 B+/+ mice). This difference in incidence of vasculitis was especially evident in female mice where 0/6 female B-/- mice had vasculitis compared to 5/7 female B+/+ mice (p ⁇ 0.01). The presence of renal vasculitis in the female B+/+ mice paralleled the vasculitis induced ear and tail necrosis present also primarily in the female B+/+ mice.
  • Immunoglobulin levels in sera or in cryoglobulins MRL/lpr renal disease and vasculitis has been linked with IgG3 cryoglobuhn production (41-44).
  • total serum immunoglobulin levels and IgG3 cryoglobuhn levels were measured by ELISA.
  • Total IgG levels were similar at all time points in all 3 groups (data not shown), however, significantly higher levels of serum IgG3 and IgG3 cryoglobulins were present in the B+/- and B+/+ mice compared to the B-/- mice (Table 3, Figure 5). Serum levels of all other IgG isotypes were not statistically different between the 3 groups (data not shown).
  • C3 levels in sera were determined by ELISA at the ages of 16, 20, and 24 weeks in all three groups. As shown in Table 4, serum C3 levels in the B-/- mice were equivalent to levels in normal C57BL6 mice and were significantly higher than serum C3 levels in the other two groups. Levels of serum C3 in the B-/- mice decreased over time suggesting some activation of C3, but were still significantly higher than the B+/+ mice even at 44 weeks of age (data not shown). These data indicate an unexpected significant role for Bf and the alternative pathway amplification loop in the consumption of C3 in this murine model of immune complex renal disease.
  • Circulating Immune Complexes To assay for effects of Bf on clearance of circulating ICs, CIC were measured by ELISA using the Clq binding assay. There were no differences in sera CIC between groups at any time point (data not shown).
  • the anti- Factor B antibody of the invention is administered parenterally, preferably intravenously, in a dosage determined in accordance with the method of the present invention.
  • An effective dosage of the antibody is that dosage required to eliminate Factor B from the serum of the subject and to eliminate the activity of Factor B in the serum.
  • the subject's vital signs, complete blood count, volume of urine output and urinary protein excretion are routinely monitored to follow the progress of treatment. Specifically, the subject is monitored for blood pressure changes that may accompany kidney disease. One sign of efficacy of treatment is lowering of the subject's elevated blood pressure.
  • the subject's red blood cell count and hematocrit are monitored to determine whether the treatment is having a beneficial effect on erythropoiesis. Because subjects with complement- mediated glomerulonephritis are often anemic, a rise in the red blood cell count and hematocrit indicate the efficacy of treatment. Further, 24-hour urinary protein excretion is measured, and a decrease in protein excretion in the urine indicates efficacy of treatment. As kidney function improves, the subject's weight decreases as edema fluid is excreted.
  • Sera IgG3 levels in B-/-, B+/+, B+/-, and MRL/lpr mice were used as a standard. Data shown are the mean IgG3 level (mg/ml) +/- standard error in each group at the age of 20 weeks. Sera IgG3 levels in B-/- were lower than the other groups. *p ⁇ 0.05 versus B-/- mice.
  • Immunoglobulin heavy chain constant region determines the pathogenicity and the antigen-binding activity of rheumatoid factor. Proc. Natl. Acad. Sci. USA 90:2345.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Medicinal Chemistry (AREA)
  • Immunology (AREA)
  • Veterinary Medicine (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Engineering & Computer Science (AREA)
  • Public Health (AREA)
  • Biophysics (AREA)
  • Environmental Sciences (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Transplantation (AREA)
  • Hematology (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne un procédé de traitement ou de prévention chez un sujet d'une maladie d'origine immunitaire induite par complément, le procédé consistant à administrer au sujet un inhibiteur de la synthèse ou de l'activité du facteur B, l'inhibition de la synthèse ou de l'activité du facteur B permettant de traiter ou de prévenir la maladie d'origine immunitaire. L'invention concerne aussi un anticorps du facteur B, un oligonucléotide antisens qui bloque l'ARNm codant pour le facteur B et un peptide qui lie concurrentiellement C3, le récepteur du facteur B. La présente invention concerne en outre un vecteur comprenant l'acide nucléique de cette invention et une cellule comprenant ce vecteur. Elle concerne ensuite un procédé d'administration à un patient de l'anticorps, de l'oligonucléotide, du peptide et du vecteur de l'invention dans un excipient pharmaceutiquement acceptable. L'invention concerne également un procédé permettant de déterminer la prédisposition d'un sujet à une maladie d'origine immunitaire induite par complément et un procédé pour déterminer l'efficacité d'un traitement et d'une prévision appliqués à un sujet souffrant d'une maladie d'origine immunitaire induite par complément. Elle concerne enfin une souris transgénique mutante.
PCT/US1999/023485 1998-10-09 1999-10-08 Facteur de blocage b destine a traiter une maladie d'origine immunitaire induite par complement WO2000021559A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU17051/00A AU1705100A (en) 1998-10-09 1999-10-08 Blocking factor b to treat complement-mediated immune disease

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US10366098P 1998-10-09 1998-10-09
US60/103,660 1998-10-09

Publications (2)

Publication Number Publication Date
WO2000021559A2 true WO2000021559A2 (fr) 2000-04-20
WO2000021559A3 WO2000021559A3 (fr) 2000-09-28

Family

ID=22296362

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1999/023485 WO2000021559A2 (fr) 1998-10-09 1999-10-08 Facteur de blocage b destine a traiter une maladie d'origine immunitaire induite par complement

Country Status (2)

Country Link
AU (1) AU1705100A (fr)
WO (1) WO2000021559A2 (fr)

Cited By (36)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1713503A1 (fr) * 2004-02-10 2006-10-25 The REGENTS OF THE UNIVERSITY OF COLORADO, A Body Corporate Inhibition du facteur b et de la voie du complement alternative et procedes associes
US7576197B2 (en) 2002-11-14 2009-08-18 Dharmacon, Inc. SiRNA targeting KRAS
US7582746B2 (en) 2002-11-14 2009-09-01 Dharmacon, Inc. siRNA targeting complement component 3 (C3)
US7592442B2 (en) 2002-11-14 2009-09-22 Dharmacon, Inc. siRNA targeting ribonucleotide reductase M2 polypeptide (RRM2 or RNR-R2)
US7605250B2 (en) 2004-05-12 2009-10-20 Dharmacon, Inc. siRNA targeting cAMP-specific phosphodiesterase 4D
US7612196B2 (en) 2002-11-14 2009-11-03 Dharmacon, Inc. siRNA targeting cyclin-dependent kinase inhibitor 1B (p27, Kip1) (CDKN1B)
US7619081B2 (en) 2002-11-14 2009-11-17 Dharmacon, Inc. siRNA targeting coatomer protein complex, subunit beta 2 (COPB2)
US7635770B2 (en) 2002-11-14 2009-12-22 Dharmacon, Inc. siRNA targeting protein kinase N-3 (PKN-3)
US7691998B2 (en) 2002-11-14 2010-04-06 Dharmacon, Inc. siRNA targeting nucleoporin 62kDa (Nup62)
US7951935B2 (en) 2002-11-14 2011-05-31 Dharmacon, Inc. siRNA targeting v-myc myelocytomatosis viral oncogene homolog (MYC)
US7959919B2 (en) * 2003-11-19 2011-06-14 Novelmed Therapeutics, Inc. Method of inhibiting factor B-mediated complement activation
US7964705B2 (en) 2007-03-14 2011-06-21 Taligen Therapeutics, Inc. Humaneered anti-factor B antibody
US7977471B2 (en) 2002-11-14 2011-07-12 Dharmacon, Inc. siRNA targeting TNFα
US7999082B2 (en) 2004-02-10 2011-08-16 National Jewish Medical And Research Center Anti-factor B antibodies
AU2006249835B2 (en) * 2005-05-26 2011-09-08 Musc Foundation For Research Development Inhibition of the alternative complement pathway for treatment of traumatic brain injury, spinal cord injury and related conditions
US8198427B1 (en) 2002-11-14 2012-06-12 Dharmacon, Inc. SiRNA targeting catenin, beta-1 (CTNNB1)
WO2013177035A3 (fr) * 2012-05-24 2014-01-16 Alexion Pharmaceuticals, Inc. Anticorps anti-facteur b humaneered
US9066925B2 (en) 2009-07-02 2015-06-30 Musc Foundation For Research Development Methods of stimulating liver regeneration
KR20160054595A (ko) * 2013-09-13 2016-05-16 아이오니스 파마수티컬즈, 인코포레이티드 보체 인자 b의 조절자
WO2017105939A1 (fr) 2015-12-16 2017-06-22 Ra Pharmaceuticals, Inc. Modulateurs de l'activité du complément
US9719092B2 (en) 2002-11-14 2017-08-01 Thermo Fisher Scientific Inc. RNAi targeting CNTD2
US9719094B2 (en) 2002-11-14 2017-08-01 Thermo Fisher Scientific Inc. RNAi targeting SEC61G
US9771586B2 (en) 2002-11-14 2017-09-26 Thermo Fisher Scientific Inc. RNAi targeting ZNF205
US9777270B2 (en) 2002-11-14 2017-10-03 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US9839649B2 (en) 2002-11-14 2017-12-12 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US9879266B2 (en) 2002-11-14 2018-01-30 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US9937222B2 (en) 2015-01-28 2018-04-10 Ra Pharmaceuticals, Inc. Modulators of complement activity
WO2018106859A1 (fr) 2016-12-07 2018-06-14 Ra Pharmaceuticals, Inc. Modulateurs de l'activité du complément
US10011836B2 (en) 2002-11-14 2018-07-03 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US10106579B2 (en) 2014-06-12 2018-10-23 Ra Pharmaceuticals, Inc. Modulation of complement activity
WO2019112984A1 (fr) 2017-12-04 2019-06-13 Ra Pharmaceuticals, Inc. Modulateurs de l'activité du complément
WO2020185541A2 (fr) 2019-03-08 2020-09-17 Ra Pharmaceuticals, Inc. Modulateurs d'activité du complément
WO2020205501A1 (fr) 2019-03-29 2020-10-08 Ra Pharmaceuticals, Inc. Modulateurs du complément et procédés associés
WO2020219822A1 (fr) 2019-04-24 2020-10-29 Ra Pharmaceuticals, Inc. Compositions et méthodes de modulation de l'activité du complément
US10961252B2 (en) 2016-06-27 2021-03-30 Achillion Pharmaceuticals, Inc. Quinazoline and indole compounds to treat medical disorders
US11732265B2 (en) 2014-05-01 2023-08-22 Ionis Pharmaceuticals, Inc. Compositions and methods for modulating complement factor B expression

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998045464A1 (fr) * 1997-04-09 1998-10-15 A+ Science Invest Ab Animal transgenique, non humain, a carence proteique, cellules derives de cet animal, utilisation dudit animal et desdites cellules, et procede de production dudit animal transgenique

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998045464A1 (fr) * 1997-04-09 1998-10-15 A+ Science Invest Ab Animal transgenique, non humain, a carence proteique, cellules derives de cet animal, utilisation dudit animal et desdites cellules, et procede de production dudit animal transgenique

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
C. CLARDY ET AL.: "In vitro inhibition of complement activation using a monoclonal antibody (MoAb) directed against human factor B (Fb)." PEDIATRIC RESEARCH, vol. 31, no. 4 part 2, 1992, page 331A XP000907515 Basel, Switzerland *
H. WATANABE ET AL.: "Amelioration of renal disease in lupus mice genetically deficient in complement factor B." ARTHRITIS & RHEUMATISM, vol. 41, no. 9 suppl., September 1998 (1998-09), page S179 XP000914560 New York, NY, USA *
J. SUNDSMO ET AL.: "Factor B, the complement alternative pathway serine proteinase, is a major constitutive protein synthesized and secreted by resident and elicited mouse macrophages." THE JOURNAL OF EXPERIMENTAL MEDICINE, vol. 161, no. 2, 1 February 1985 (1985-02-01), pages 306-322, XP000907493 New York, NY, USA *
K. TRAUSTADOTTIR ET AL.: "Participation of factor B in residual immune complex red cell binding activity observed in serum from a C2-deficient systemic lupus erythematosus patient may delay the appearance of clinical symptoms." ARTHRITIS & RHEUMATISM, vol. 41, no. 3, March 1998 (1998-03), pages 427-434, XP000907526 New York, NY, USA *
M. FRANK: "Animal models for complement deficiencies." JOURNAL OF CLINICAL IMMUNOLOGY, vol. 15, no. 6 suppl., November 1995 (1995-11), pages 113S-121S, XP000907494 New York, NY, USA *
S. MIYAGAWA ET AL.: "Prolonging discordant xenograft survival with anticomplement reagents K76COOH and FUT175." TRANSPLANTATION, vol. 55, no. 4, April 1993 (1993-04), pages 709-713, XP000907527 Baltimore, MD, USA *

Cited By (108)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8222395B2 (en) 2002-11-14 2012-07-17 Dharmacon, Inc. siRNA targeting kinase insert domain receptor (KDR)
US7635770B2 (en) 2002-11-14 2009-12-22 Dharmacon, Inc. siRNA targeting protein kinase N-3 (PKN-3)
US7576197B2 (en) 2002-11-14 2009-08-18 Dharmacon, Inc. SiRNA targeting KRAS
US7576196B2 (en) 2002-11-14 2009-08-18 Dharmacon, Inc. siRNA targeting transducin (beta)-like 3 (TBL3)
US7582746B2 (en) 2002-11-14 2009-09-01 Dharmacon, Inc. siRNA targeting complement component 3 (C3)
US7589191B2 (en) 2002-11-14 2009-09-15 Dharmacon, Inc. siRNA targeting hypoxia-inducible factor 1
US7592442B2 (en) 2002-11-14 2009-09-22 Dharmacon, Inc. siRNA targeting ribonucleotide reductase M2 polypeptide (RRM2 or RNR-R2)
US7592443B2 (en) 2002-11-14 2009-09-22 Dharmacon, Inc. siRNA targeting interleukin-1 receptor-associated kinase 4 (IRAK4)
US7592444B2 (en) 2002-11-14 2009-09-22 Dharmacon, Inc. siRNA targeting myeloid cell leukemia sequence 1
US7595389B2 (en) 2002-11-14 2009-09-29 Dharmacon, Inc. siRNA targeting casitas B cell lymphoma-B (CBL-B)
US7598370B2 (en) 2002-11-14 2009-10-06 Dharmacon, Inc. siRNA targeting polo-like kinase-1 (PLK-1)
US7598369B2 (en) 2002-11-14 2009-10-06 Dharmacon, Inc. siRNA targeting histamine receptor H1
US7605252B2 (en) 2002-11-14 2009-10-20 Dharmacon, Inc. siRNA targeting kinase insert domain receptor (KDR)
US11198870B2 (en) 2002-11-14 2021-12-14 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US7608707B2 (en) 2002-11-14 2009-10-27 Dharmacon, Inc. siRNA targeting survivin
US7612196B2 (en) 2002-11-14 2009-11-03 Dharmacon, Inc. siRNA targeting cyclin-dependent kinase inhibitor 1B (p27, Kip1) (CDKN1B)
US7615541B2 (en) 2002-11-14 2009-11-10 Dharmacon, Inc. siRNA targeting TIE-2
US7619081B2 (en) 2002-11-14 2009-11-17 Dharmacon, Inc. siRNA targeting coatomer protein complex, subunit beta 2 (COPB2)
US7632938B2 (en) 2002-11-14 2009-12-15 Dharmacon, Inc. siRNA targeting superoxide dismutase 1 (SOD1)
US7632939B2 (en) 2002-11-14 2009-12-15 Dharmacon, Inc. siRNA targeting proto-oncogene MET
US10765695B2 (en) 2002-11-14 2020-09-08 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US7635771B2 (en) 2002-11-14 2009-12-22 Dharmacon, Inc. siRNA targeting amyloid beta (A4) precursor protein (APP)
US7642349B2 (en) 2002-11-14 2010-01-05 Dharmacon, Inc. siRNA targeting TATA box binding protein (TBP)-associated factor (TAF1)
US7655789B2 (en) 2002-11-14 2010-02-02 Dharmacon, Inc. siRNA targeting transient receptor potential cation channel, subfamily V, member 1 (TRPV1)
US7666853B2 (en) 2002-11-14 2010-02-23 Dharmacon, Inc. siRNA targeting connective tissue growth factor (CTGF)
US7678896B2 (en) 2002-11-14 2010-03-16 Dharmacon, Inc. siRNA targeting serine/threonine kinase 12 (STK12 or aurora B kinase)
US7691998B2 (en) 2002-11-14 2010-04-06 Dharmacon, Inc. siRNA targeting nucleoporin 62kDa (Nup62)
US7691997B2 (en) 2002-11-14 2010-04-06 Dharmacon, Inc. Functional and hyperfunctional siRNA
US7696344B2 (en) * 2002-11-14 2010-04-13 Dharmacon, Inc. siRNA targeting complement factor B
US7709629B2 (en) 2002-11-14 2010-05-04 Dharmacon, Inc. siRNA targeting diacylglycerol O-acyltransferase homolog 2 (DGAT2)
US7737267B2 (en) 2002-11-14 2010-06-15 Dharmacon, Inc. siRNA targeting hypoxia-inducible factor 1
US8198427B1 (en) 2002-11-14 2012-06-12 Dharmacon, Inc. SiRNA targeting catenin, beta-1 (CTNNB1)
US7745612B2 (en) 2002-11-14 2010-06-29 Dharmacon, Inc. siRNA targeting interleukin-1 receptor-associated kinase 4 (IRAK4)
US7745611B2 (en) 2002-11-14 2010-06-29 Dharmacon, Inc. siRNA targeting KRAS
US7745610B2 (en) 2002-11-14 2010-06-29 Dharmacon, Inc. siRNA targeting cyclin dependent kinase 11 (CDK11)
US7795420B2 (en) 2002-11-14 2010-09-14 Dharmacon, Inc. Functional and hyperfunctional siRNA directed against Bcl-2
US7807819B2 (en) 2002-11-14 2010-10-05 Dharmacon, Inc. siRNA targeting survivin
US7820809B2 (en) 2002-11-14 2010-10-26 Dharmacon, Inc. Functional and hyperfunctional siRNA directed against Bcl-2
US7833989B2 (en) 2002-11-14 2010-11-16 Dharmacon, Inc. siRNA targeting connective tissue growth factor (CTGF)
US7855186B2 (en) 2002-11-14 2010-12-21 Dharmacon, Inc. siRNA targeting TIE-2
US7897754B2 (en) 2002-11-14 2011-03-01 Dharmacon, Inc. SiRNA targeting ras-related nuclear protein RAN
US7951935B2 (en) 2002-11-14 2011-05-31 Dharmacon, Inc. siRNA targeting v-myc myelocytomatosis viral oncogene homolog (MYC)
US10696968B2 (en) 2002-11-14 2020-06-30 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US10233449B2 (en) 2002-11-14 2019-03-19 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US8039610B2 (en) 2002-11-14 2011-10-18 Dharmacon, Inc. siRNA targeting superoxide dismutase 1 (SOD1)
US10011836B2 (en) 2002-11-14 2018-07-03 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US7977471B2 (en) 2002-11-14 2011-07-12 Dharmacon, Inc. siRNA targeting TNFα
US9879266B2 (en) 2002-11-14 2018-01-30 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US8013145B2 (en) 2002-11-14 2011-09-06 Dharmacon, Inc. SiRNA targeting cyclin-dependent kinase inhibitor 1B (p27, Kip1) (CDKN1B)
US9839649B2 (en) 2002-11-14 2017-12-12 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US8022199B2 (en) 2002-11-14 2011-09-20 Dharmacon, Inc. SiRNA targeting myeloid differentiation primary response gene (88) (MYD88)
US8022198B2 (en) 2002-11-14 2011-09-20 Dharmacon, Inc. siRNA targeting histamine receptor H1
US9777270B2 (en) 2002-11-14 2017-10-03 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US7741470B2 (en) 2002-11-14 2010-06-22 Dharmacon, Inc. siRNA targeting gremlin
US9771586B2 (en) 2002-11-14 2017-09-26 Thermo Fisher Scientific Inc. RNAi targeting ZNF205
US8304528B2 (en) 2002-11-14 2012-11-06 Dharmacon, Inc. SiRNA targeting fructose-1, 6-bisphosphatase 1 (FBP1)
US9719094B2 (en) 2002-11-14 2017-08-01 Thermo Fisher Scientific Inc. RNAi targeting SEC61G
US9719092B2 (en) 2002-11-14 2017-08-01 Thermo Fisher Scientific Inc. RNAi targeting CNTD2
US7959919B2 (en) * 2003-11-19 2011-06-14 Novelmed Therapeutics, Inc. Method of inhibiting factor B-mediated complement activation
US7999082B2 (en) 2004-02-10 2011-08-16 National Jewish Medical And Research Center Anti-factor B antibodies
EP1713503A4 (fr) * 2004-02-10 2008-08-06 Univ Colorado Regents Inhibition du facteur b et de la voie du complement alternative et procedes associes
EP1713503A1 (fr) * 2004-02-10 2006-10-25 The REGENTS OF THE UNIVERSITY OF COLORADO, A Body Corporate Inhibition du facteur b et de la voie du complement alternative et procedes associes
EP2340850A1 (fr) * 2004-02-10 2011-07-06 The Regents of the University of Colorado, a Body Corporate Inhibition du facteur B et de la voie du complément alternative et procédés associés
AU2005212369B2 (en) * 2004-02-10 2011-06-23 Musc Foundation For Research Development Inhibition of factor B, the alternative complement pathway and methods related thereto
US8652475B2 (en) 2004-02-10 2014-02-18 Musc Foundation For Research Development Inhibition of factor B, the alternative complement pathway and methods related thereto
US8703140B2 (en) 2004-02-10 2014-04-22 Musc Foundation For Research Development Inhibition of factor B, the alternative complement pathway and methods related thereto
US7605250B2 (en) 2004-05-12 2009-10-20 Dharmacon, Inc. siRNA targeting cAMP-specific phosphodiesterase 4D
AU2006249835B2 (en) * 2005-05-26 2011-09-08 Musc Foundation For Research Development Inhibition of the alternative complement pathway for treatment of traumatic brain injury, spinal cord injury and related conditions
US8911733B2 (en) 2005-05-26 2014-12-16 Musc Foundation For Research Development Inhibition of the alternative complement pathway for treatment of traumatic brain injury, spinal cord injury and related conditions
US9096677B2 (en) 2007-03-14 2015-08-04 Alexion Pharmaceuticals, Inc. Humaneered anti-factor B antibody
US7964705B2 (en) 2007-03-14 2011-06-21 Taligen Therapeutics, Inc. Humaneered anti-factor B antibody
US9066925B2 (en) 2009-07-02 2015-06-30 Musc Foundation For Research Development Methods of stimulating liver regeneration
US9803005B2 (en) 2012-05-24 2017-10-31 Alexion Pharmaceuticals, Inc. Humaneered anti-factor B antibody
WO2013177035A3 (fr) * 2012-05-24 2014-01-16 Alexion Pharmaceuticals, Inc. Anticorps anti-facteur b humaneered
KR102381790B1 (ko) * 2013-09-13 2022-03-31 아이오니스 파마수티컬즈, 인코포레이티드 보체 인자 b의 조절자
KR20160054595A (ko) * 2013-09-13 2016-05-16 아이오니스 파마수티컬즈, 인코포레이티드 보체 인자 b의 조절자
US11732265B2 (en) 2014-05-01 2023-08-22 Ionis Pharmaceuticals, Inc. Compositions and methods for modulating complement factor B expression
US11014965B2 (en) 2014-06-12 2021-05-25 Ra Pharmaceuticals, Inc. Modulation of complement activity
US10106579B2 (en) 2014-06-12 2018-10-23 Ra Pharmaceuticals, Inc. Modulation of complement activity
US10435438B2 (en) 2014-06-12 2019-10-08 Ra Pharmaceuticals, Inc. Modulation of complement activity
US10562934B2 (en) 2014-06-12 2020-02-18 Ra Pharmaceuticals, Inc. Modulation of complement activity
US11965040B2 (en) 2014-06-12 2024-04-23 Ra Pharmaceuticals, Inc. Modulation of complement activity
EP4223317A2 (fr) 2014-06-12 2023-08-09 RA Pharmaceuticals, Inc. Modulation de l'activité du complément
EP3628680A1 (fr) 2014-06-12 2020-04-01 RA Pharmaceuticals, Inc. Modulation d'activité complémentaire
US11535650B1 (en) 2014-06-12 2022-12-27 Ra Pharmaceuticals, Inc. Modulation of complement activity
US10208089B2 (en) 2014-06-12 2019-02-19 Ra Pharmaceuticals, Inc. Modulation of complement activity
EP3973994A1 (fr) 2014-06-12 2022-03-30 RA Pharmaceuticals, Inc. Modulation d'activité complémentaire
US9937222B2 (en) 2015-01-28 2018-04-10 Ra Pharmaceuticals, Inc. Modulators of complement activity
US12239684B2 (en) 2015-01-28 2025-03-04 Ra Pharmaceuticals, Inc. Modulators of complement activity
US11707503B2 (en) 2015-01-28 2023-07-25 Ra Pharmaceuticals, Inc. Modulators of complement activity
US10918691B2 (en) 2015-01-28 2021-02-16 Ra Pharmaceuticals, Inc. Modulators of complement activity
EP3988110A1 (fr) 2015-01-28 2022-04-27 RA Pharmaceuticals, Inc. Modulateurs d'activité complémentaire
US10328115B2 (en) 2015-01-28 2019-06-25 Ra Pharmaceuticals, Inc. Modulators of complement activity
US10588936B2 (en) 2015-01-28 2020-03-17 Ra Pharmaceuticals, Inc. Modulators of complement activity
WO2017105939A1 (fr) 2015-12-16 2017-06-22 Ra Pharmaceuticals, Inc. Modulateurs de l'activité du complément
US11752190B2 (en) 2015-12-16 2023-09-12 Ra Pharmaceuticals, Inc. Modulators of complement activity
EP3685847A1 (fr) 2015-12-16 2020-07-29 RA Pharmaceuticals, Inc. Modulateurs d'activité complémentaire
EP4218790A1 (fr) 2015-12-16 2023-08-02 Ra Pharmaceuticals, Inc. Modulateurs de l'activité du complément
US10835574B2 (en) 2015-12-16 2020-11-17 Ra Pharmaceuticals, Inc. Modulators of complement activity
US11248000B2 (en) 2016-06-27 2022-02-15 Achillion Pharmaceuticals, Inc. Quinazoline and indole compounds to treat medical disorders
US10961252B2 (en) 2016-06-27 2021-03-30 Achillion Pharmaceuticals, Inc. Quinazoline and indole compounds to treat medical disorders
US11123399B2 (en) 2016-12-07 2021-09-21 Ra Pharmaceuticals, Inc. Modulators of complement activity
US11723949B2 (en) 2016-12-07 2023-08-15 Ra Pharmaceuticals, Inc. Modulators of complement activity
WO2018106859A1 (fr) 2016-12-07 2018-06-14 Ra Pharmaceuticals, Inc. Modulateurs de l'activité du complément
WO2019112984A1 (fr) 2017-12-04 2019-06-13 Ra Pharmaceuticals, Inc. Modulateurs de l'activité du complément
WO2020185541A2 (fr) 2019-03-08 2020-09-17 Ra Pharmaceuticals, Inc. Modulateurs d'activité du complément
WO2020205501A1 (fr) 2019-03-29 2020-10-08 Ra Pharmaceuticals, Inc. Modulateurs du complément et procédés associés
WO2020219822A1 (fr) 2019-04-24 2020-10-29 Ra Pharmaceuticals, Inc. Compositions et méthodes de modulation de l'activité du complément

Also Published As

Publication number Publication date
AU1705100A (en) 2000-05-01
WO2000021559A3 (fr) 2000-09-28

Similar Documents

Publication Publication Date Title
WO2000021559A2 (fr) Facteur de blocage b destine a traiter une maladie d'origine immunitaire induite par complement
Sekine et al. Complement component C3 is not required for full expression of immune complex glomerulonephritis in MRL/lpr mice
Quigg et al. Transgenic mice overexpressing the complement inhibitor crry as a soluble protein are protected from antibody-induced glomerular injury
Wang et al. Mechanism of action of combined short-term CTLA4Ig and anti-CD40 ligand in murine systemic lupus erythematosus
Chen et al. Humoral immune responses in Cr2−/− mice: enhanced affinity maturation but impaired antibody persistence
US20020138863A1 (en) FcRn-based therapeutics for the treatment of auto-immune disorders
Merino et al. Prevention of systemic lupus erythematosus in autoimmune BXSB mice by a transgene encoding IE alpha chain.
JP4708510B2 (ja) サイトカイン媒介毒性に関係する疾患の治療における遊走阻止因子の抑制
US20070123466A1 (en) Method of treating recurrent miscarriages
Ait-Azzouzene et al. An immunoglobulin Cκ-reactive single chain antibody fusion protein induces tolerance through receptor editing in a normal polyclonal immune system
US20070059758A1 (en) Regulators of type-1 tumor necrosis factor receptor and other cytokine receptor shedding
EP1368458A2 (fr) Gene induit par l'interferon alpha
UA127339C2 (uk) СПОСІБ ПРОФІЛАКТИКИ АБО ЗМЕНШЕННЯ УРАЖЕННЯ НИРОК У СУБ'ЄКТА, ЩО СТРАЖДАЄ НА СТЕРОЇДЗАЛЕЖНУ ІМУНОГЛОБУЛІН-А-НЕФРОПАТІЮ (IgAN)
Sarkar et al. Partial correction of murine hemophilia A with neo-antigenic murine factor VIII
WO2001064749A2 (fr) Procede de preparation d'anticorps anti-facteur d'inhibition de la migration des macrophages
CN101932337A (zh) 替代途径的血清灭菌蛋白调节及其应用
US20150139999A1 (en) Interferon antagonists, antibodies thereto, and associated methods of use
Pollard et al. Costimulation requirements of induced murine systemic autoimmune disease
JP2001513822A (ja) 免疫性凝固を調節する方法
US8084415B2 (en) Uteroglobin in the treatment of IGA mediated nephropathy
Muraoka et al. IK cytokine ameliorates the progression of lupus nephritis in MRL/lpr mice
US20100015654A1 (en) Negative regulation of NK cell functions by EAT-2, a sap-related adaptor expressed in innate immune cells
JP2004500872A (ja) Ｇｐ３５４核酸およびポリペプチド
Holers et al. Complement Component C3 Is Not Required
AU2001272982B2 (en) GP286 nucleic acids and polypeptides

Legal Events

Date Code Title Description
ENP Entry into the national phase in:

Ref country code: AU

Ref document number: 2000 17051

Kind code of ref document: A

Format of ref document f/p: F

AK Designated states

Kind code of ref document: A2

Designated state(s): AU CA JP US

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
AK Designated states

Kind code of ref document: A3

Designated state(s): AU CA JP US

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

122 Ep: pct application non-entry in european phase
点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载