Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
  • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
  • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
• • 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
  • 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
  • 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
• • 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
• • 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
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2851—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the lectin superfamily, e.g. CD23, CD72
• • 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
  • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule

Definitions

• the present application relates generally to pharmaceutical compositions and dosage regimens for the clinical use of anti-Blood Dendritic Cell Antigen 2 antibodies.
• Blood dendritic cell antigen 2 is a C-type lectin expressed on human plasmacytoid dendritic cells (pDCs) (Dzionek et al., J. Immunol., 165:6037-6046 (2000)), a specialized population of bone marrow-derived cells that secrete type I interferons (IFNs) in response to toll-like receptor (TLR) ligands.
• BDCA2 consists of a single extracellular carbohydrate recognition domain (CRD), which belongs to the type ⁇ C-type lectin group, at its C-terminus, a transmembrane region, and a short cytoplasmic tail at its N- terminus that does not harbor a signaling motif.
• BDCA2 transmits intracellular signals through an associated transmembrane adaptor, the FceRIy, and induces a B cell receptor (BCR)-like signaling cascade.
• BCR B cell receptor
• compositions and dosage regimens of anti-BDCA2 antibodies or BDCA2-binding fragments thereof relate, in part, to compositions and dosage regimens of anti-BDCA2 antibodies or BDCA2-binding fragments thereof and their use in the treatment of BDCA2- associated disorders such as systematic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), and discoid lupus erythematosus (DLE).
• SLE systematic lupus erythematosus
• CLE cutaneous lupus erythematosus
• DLE discoid lupus erythematosus
• the disclosure features a pharmaceutical composition
• a pharmaceutical composition comprising an anti- BDCA2 antibody or BDCA2-binding fragment thereof, sucrose, and arginine hydrochloride (ArgJHCl).
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL comprising the CDRs of BIIB059.
• VH immunoglobulin heavy chain variable domain
• VL immunoglobulin light chain variable domain
• the six CDRs of ⁇ 059 comprise or consist of the amino acid sequences set forth in SEQ ID NO: 1 or 17; SEQ ID NO:2; SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:5; and SEQ 1D N0:6.
• the composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 50 mg/ml to 225 mg/ml . In other embodiments, the composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 125 mg/ml to 175 mg/ml. In certain embodiments, the composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 150 mg/ml.
• the composition comprises sucrose at a concentration of 0.05% to 10%. In other embodiments, the composition comprises sucrose at a concentration of 1% to 5%. In certain embodiments, the composition comprises sucrose at a concentration of 3%.
• the composition comprises Arg.HCl at a concentration of 50 mM to 250 mM. In other embodiments, the composition comprises Arg.HCl at a concentration of 75 mM to 125 mM. In certain embodiments, the composition comprises Arg.HCl at a concentration of 100 mM.
• the composition further comprises Polysorbate-80 (PS80).
• PS80 Polysorbate-80
• the composition comprises PS80 at a concentration of 0.01% to 0.1%.
• the composition comprises PS80 at a concentration of 0.03% to 0.08%.
• the composition comprises PS80 at a concentration of 0.05%.
• the composition further comprises histidine. In some embodiments, the composition comprises histidine at a concentration of 5 mM to 50 mM. In other embodiments, the composition comprises histidine at a concentration of 15 mM to 25 mM. In certain embodiments, the composition comprises histidine at a concentration of 20 mM.
• the composition has a pH of 5.3 to 5.7. In other embodiments, the composition has a pH of 5.5.
• the composition further comprises methionine. In some embodiments, the composition comprises methionine at a concentration of 1 mM to 20 mM. In other embodiments, the composition comprises methionine at a concentration of 5 mM to 15 mM. In certain embodiments, the composition comprises methionine at a concentration of 10 mM.
• the composition further comprises glutamic acid. In some embodiments, the composition comprises glutamic acid at a concentration of 50 mM to 100 mM. In other embodiments, the composition comprises glutamic acid at a concentration of 50 mM to 80 mM. In certain embodiments, the composition comprises glutamic acid at a concentration of 70 mM.
• the pharmaceutical composition comprises the anti-BDCA2 antibody or the BDCA2-binding fragment thereof at a concentration of 125 mg/ml to 175 mg/ml; sucrose at a concentration of 1% to 5%; histidine at a concentration of 15 mM to 25 mM; Arg.HCl at a concentration of 75 mM to 125 mM; and PS80 at a concentration of 0.03% to 0.08%.
• the composition has a pH of 5.3 to 5.7.
• the composition also comprises methionine at a concentration of 5 mM to 15 mM.
• the composition also comprises glutamic acid at a concentration of 60 mM to 80 mM.
• the pharmaceutical composition comprises the anti-BDCA2 antibody or the BDCA2-binding fragment thereof at a concentration of 150 mg/ml; sucrose at a concentration of 3%; histidine at a concentration of 20 mM; Arg.HCl at a concentration of 100 mM; and PS80 at a concentration of 0.05%.
• the composition has a pH of 5.5.
• the composition also comprises methionine at a concentration of 10 mM.
• the composition also comprises glutamic acid at a concentration of 70 mM.
• the VH comprises or consists of a sequence at least 80% identical to SEQ ID NO:7 and the VL comprises or consists of a sequence at least 80% identical to SEQ ID NO:8. In some embodiments, the VH comprises or consists of a sequence at least 90% identical to SEQ ID NO:7 and the VL comprises or consists of a sequence at least 90% identical to SEQ ID NO:8. In some embodiments, the VH comprises or consists of the sequence of SEQ ID NO:7 and the VL comprises or consists of the sequence ofSEQ ID NO:8.
• the anti-BDCA2 antibody comprises an immunoglobulin heavy chain and an immunoglobulin light chain.
• the heavy chain comprises or consists of a sequence at least 80% identical to SEQ ID NO:9 and the light chain comprises or consists of a sequence at least 80% identical to SEQ ID NO: 10.
• the heavy chain comprises or consists of a sequence at least 90% identical to SEQ ID NO:9 and the light chain comprises or consists of a sequence at least 90% identical to SEQ ID NO: 10.
• the heavy- chain comprises or consists of the sequence of SEQ ID NO:9 and the light chain comprises or consists of the sequence of SEQ ID NO: 10.
• the disclosure features a method of treating a condition selected from the group consisting of systemic lupus erythematosus, cutaneous lupus erythematosus, discoid lupus erythematosus, Sjogren's syndrome, dermatopolymyositis, scleroderma, and cytokine release syndrome in a human subject in need thereof.
• the method involves administering to the human subject a pharmaceutical composition described herein.
• the pharmaceutical composition is administered
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof of the pharmaceutical composition is administered to the human subject at a dose of 50 mg every four weeks.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof of the pharmaceutical composition is administered to the human subject at a dose of 150 mg every four weeks.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof of the pharmaceutical composition is administered to the human subject at a dose of 450 mg every four weeks.
• the disclosure provides a method of treating a condition selected from the group consisting of systemic lupus erythematosus, cutaneous lupus erythematosus, discoid lupus erythematosus, Sjogren's syndrome, dermatopolymyositis, scleroderma, and cytokine release syndrome in a human subject in need thereof.
• the method comprises administering subcutaneously to the human subject an anti-BDCA2 antibody or BDCA2- binding fragment thereof at a dose of 50 mg every four weeks.
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL).
• VH and VL respectively, comprise:
• H-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 ;
• H-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2;
• H-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3;
• L-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4
• L-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5
• L-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• the human subject is administered a loading dose of the anti-
• the disclosure provides a method of treating a condition selected from the group consisting of systemic lupus erythematosus, cutaneous lupus erythematosus, discoid lupus erythematosus, Sjogren's syndrome, dermatopolymyositis, scleroderma, and cytokine release syndrome in a human subject in need thereof.
• the method comprises administering subcutaneously to the human subject an anti-BDCA2 antibody or BDCA2- binding fragment thereof at a dose of 150 mg every four weeks.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL).
• VH and VL respectively, comprise:
• H-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 ;
• H-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2;
• H-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3;
• L-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4
• L-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5
• L-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• the human subject is administered a loading dose of the anti- BDCA2 antibody or BDCA2-binding fragment thereof two weeks after the first
• the loading dose is 150 mg.
• the disclosure provides a method of treating a condition selected from the group consisting of systemic lupus erythematosus, cutaneous lupus erythematosus, discoid lupus erythematosus, Sjogren's syndrome, dermatopolymyositis, scleroderma, and cytokine release syndrome in a human subject in need thereof.
• the method comprises administering subcutaneously to the human subject an anti-BDCA2 antibody or BDCA2- binding fragment thereof at a dose of 450 mg every four weeks.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL).
• VH and VL comprise: VH complementarity determining regions (CDRs), wherein H-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1; H-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and H-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein L-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; L CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and L-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• CDRs VH complementarity determining regions
• the human subject is administered a loading dose of the anti- BDCA2 antibody or BDCA2-binding fragment thereof two weeks after the first
• the loading dose is 450 mg.
• the human subject is administered at least 4 doses of the anti-BDCA2 antibody or antigen-binding fragment thereof. In some embodiments, the human subject is administered at least 7 doses of the anti-BDCA2 antibody or antigen-binding fragment thereof. In certain embodiments, the human subject is administered at least 10 doses of the anti-BDCA2 antibody or antigen-binding fragment thereof.
• the VH comprises or consists of a sequence at least 80% identical to SEQ ID NO:7 and the VL comprises or consists of a sequence at least 80% identical to SEQ ID NO:8.
• the VH comprises or consists of a sequence at least 90% identical to SEQ ID NO:7 and the VL comprises or consists of a sequence at least 90% identical to SEQ ID NO:8. In some embodiments, the VH comprises or consists of the sequence of SEQ ID NO:7 and the VL comprises or consists of the sequence of SEQ ID NO:8.
• the anti-BDCA2 antibody comprises an immunoglobulin heavy chain and an immunoglobulin light chain. In certain instances, the heavy chain comprises or consists of a sequence at least 80% identical to SEQ ID NO:9 and the light chain comprises or consists of a sequence at least 80% identical to SEQ ID NO: 10.
• the heavy- chain comprises or consists of a sequence at least 90% identical to SEQ ID NO:9 and the light chain comprises or consists of a sequence at least 90% identical to SEQ ID NO: 10. In yet other instances, the heavy chain comprises or consists of the sequence of SEQ ID NO:9 and the light chain comprises or consists of the sequence of SEQ ID NO: 10.
• the condition is systemic lupus erythematosus. In other embodiments, the condition is cutaneous lupus erythematosus (with or without SLE). In some embodiments, the condition is discoid lupus erythematosus (with or without SLE). In certain embodiments, the condition is cytokine release syndrome.
• the disclosure features a syringe, injector (e.g., autoinjector, subcutaneous large volume injector), or pump comprising a sterile preparation of the pharmaceutical composition described herein adapted for subcutaneous administration of the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a fixed dose of 50 mg, 150 mg, or 450 mg.
• injector e.g., autoinjector, subcutaneous large volume injector
• pump comprising a sterile preparation of the pharmaceutical composition described herein adapted for subcutaneous administration of the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a fixed dose of 50 mg, 150 mg, or 450 mg.
• the disclosure provides a syringe, injector, or pump comprising a sterile preparation of an anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the syringe or pump is adapted for subcutaneous administration of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof at a fixed dose of 50 mg, 150 mg, or 450 mg.
• the anti- BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL).
• VH and VL comprise: VH complementarity detennining regions (CDRs), wherein H-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1; H-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and H-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein L-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; L CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and L-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• CDRs VH complementarity detennining regions
• the VH comprises or consists of a sequence at least 80% identical to SEQ ID NO:7 and the VL comprises or consists of a sequence at least 80% identical to SEQ ID NO:8. In some embodiments, the VH comprises or consists of a sequence at least 90% identical to SEQ ID NO:7 and the VL comprises or consists of a sequence at least 90% identical to SEQ ID NO:8. In some embodiments, the VH comprises or consists of the sequence of SEQ ID NO:7 and the VL comprises or consists of the sequence of SEQ ID NO:8. In some embodiments, the anti-BDCA2 antibody comprises an immunoglobulin heavy chain and an immunoglobulin light chain.
• the heavy chain comprises or consists of a sequence at least 80% identical to SEQ ID NO:9 and the light chain comprises or consists of a sequence at least 80% identical to SEQ ID NO: 10. In other instances, the heavy chain comprises or consists of a sequence at least 90% identical to SEQ ID NO:9 and the light chain comprises or consists of a sequence at least 90% identical to SEQ ID NO: 10. In yet other instances, the heavy chain comprises or consists of the sequence of SEQ ID NO:9 and the light chain comprises or consists of the sequence of SEQ ID NO: 10.
• the disclosure provides a pharmaceutical composition
• a pharmaceutical composition comprising an anti-BDCA2 antibody or BDCA2-binding fragment thereof, sucrose, and arginine hydrochloride (Arg.HCl), wherein the pharmaceutical composition has a pH of 5.0 to 6.5.
• sucrose is not part of the pharmaceutical composition.
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL comprising the CDRs of ⁇ 059.
• the six CDRs of BIIB059 comprise or consist of the amino acid sequences set forth in SEQ ID NO: 1 or 17; SEQ ID NO:2; SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:5; and SEQ ID NO:6.
• the pharmaceutical composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 50 mg/ml to 225 mg/ml. In some embodiments, the pharmaceutical composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 125 mg/ml to 175 mg/ml. In other embodiments, the pharmaceutical composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 150 mg/ml. In certain embodiments, the pharmaceutical composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 200 mg/ml. In certain embodiments, the
• composition comprises the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a concentration of 225 mg/ml.
• the pharmaceutical composition comprises sucrose at a concentration of 1% to 10%. In some embodiments, the pharmaceutical composition comprises sucrose at a concentration of 1% to 5%. In certain embodiments, the
• composition comprises sucrose at a concentration of 1%. In certain embodiments, the pharmaceutical composition comprises sucrose at a concentration of 3%.
• the composition comprises Arg.HCl at a concentration of 50 oiM to 250 mM. In some embodiments, the composition comprises Arg.HCl at a concentration of 50 mM to 200 mM. In other embodiments, the composition comprises Arg.HCl at a concentration of 75 mM to 150 mM. In other embodiments, the composition comprises Arg.HCl at a concentration of 75 mM to 125 mM. In some embodiments, the composition comprises Arg.HCl at a concentration of 100 mM to 250 mM. In some embodiments, the composition comprises Arg.HCl at a concentration of 100 mM to 200 mM. In certain embodiments, the composition comprises Arg.HCl at a concentration of 100 mM. In certain embodiments, the composition comprises Arg.HCl at a concentration of 250 mM.
• the pharmaceutical composition comprises polysorbate-80. In certain instances, the composition comprises PS80 at a concentration of 0.02% to 0.08%. In other instances, the composition comprises PS80 at a concentration of 0.03% to 0.08%. In yet other instances, the composition comprises PS80 at a concentration of 0.05%. In some embodiments, the pharmaceutical composition comprises histidine. In certain instances, the composition comprises histidine at a concentration of 10 raM to 30 mM. In other instances, the composition comprises histidine at a concentration of 15 mM to 25 mM. In yet other instances, the composition comprises histidine at a concentration of 20 mM.
• the pharmaceutical composition has a pH of5.3 to 6.5. In certain instances, the composition has a pH of 5.3 to 6.0. In certain instances, the composition has a pH of 5.5. In certain instances, the composition has a pH of 6.0.
• the pharmaceutical composition comprises athiol-containing antioxidant.
• the thiol-containing antioxidant is GSH, GSSG, the combination of GSH and GSSG, cystine, cysteine, or the combination of cysteine and cystine.
• the thiol-containing antioxidant is GSH.
• the thiol- containing antioxidant is GSSG.
• the thiol-containing antioxidant is the combination of GSH and GSSG.
• the thiol-containing antioxidant is cysteine.
• the thiol-containing antioxidant is the combination of cysteine and cystine.
• the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.02 mM to 2 mM. In some instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.2 mM. In other instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.4 mM. In some instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 1.0 mM. In certain cases, GSH and GSSG are found in the pharmaceutical composition at concentrations of 0.4 mM and 0.2 mM, respectively. In other cases, cysteine and cystine are found in the pharmaceutical composition at concentrations of 0.4 mM and 0.2 mM, respectively.
• the disclosure provides a pharmaceutical composition
• a pharmaceutical composition comprising an anti-Blood Dendritic Cell Antigen 2 (BDCA2) antibody or BDCA2-binding fragment thereof and histidine at a concentration of 10 mM to 30 mM, Arg.HCl at a concentration of 50 mM to 250 mM, and PS80 at a concentration of 0.02% to 0.08%, wherein the composition has apH of5.0 to 6.5.
• BDCA2 anti-Blood Dendritic Cell Antigen 2
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID N0:3; and VL CDRs, wherein VL-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17
• the pharmaceutical composition has an anti-BDCA2 antibody or the BDCA2 -binding fragment thereof at a concentration of 50 mg/ml to 225 mg/ml.
• the pharmaceutical composition comprises sucrose at a concentration of l%to 10%.
• the pharmaceutical composition comprises a thiol-containing antioxidant.
• the thiol-containing antioxidant is GSH, GSSG, the combination of GSH and GSSG, cystine, cysteine, or the combination of cysteine and cystine.
• the thiol-containing antioxidant is GSH.
• the thiol- containing antioxidant is GSSG.
• the thiol-containing antioxidant is the combination of GSH and GSSG.
• the thiol-containing antioxidant is cysteine.
• the thiol-containing antioxidant is the combination of cysteine and cystine.
• the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.02 niM to 2 mM. In some instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.2 mM. In other instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.4 mM. In some instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 1.0 mM. In certain cases, GSH and GSSG are found in the pharmaceutical composition at concentrations of 0.4 mM and 0.2 mM, respectively. In other cases, cysteine and cystine are found in the pharmaceutical composition at concentrations of 0.4 mM and 0.2 mM, respectively.
• the pharmaceutical composition comprises the anti-BDCA2 antibody or the BDCA2-binding fragment thereof at a concentration of 150 mg/ml, sucrose at a concentration of 3%, histidine at a concentration of 20 mM, Arg.HCl at a concentration of 100 mM, PS80 at a concentration of 0.05%, and GSH or cysteine at a concentration of 0.4 mM.
• the composition has a pH of 5.5.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH- CDR1 consists of the amino acid sequence set forth in SEQ ID NO:l or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• sucrose is not part of this composition.
• the pharmaceutical composition comprises the anti-BDCA2 antibody or the BDCA2-binding fragment thereof at a concentration of 150 mg/ml, sucrose at a concentration of 3%, histidine at a concentration of 20 mM, Arg.HCl at a concentration of 100 mM, PS80 at a concentration of 0.05%, and GSSG or cystine at a concentration of 0.2 mM.
• the composition has a pH of 5.5.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH- CDR1 consists of the amino acid sequence set forth in SEQ ID NO:l or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• sucrose is not part of this composition.
• the pharmaceutical composition comprises the anti- BDCA2 antibody or the BDCA2 -binding fragment thereof at a concentration of 150 mg/ml, sucrose at a concentration of 3%, histidine at a concentration of 20 mM, Arg.HCl at a concentration of 100 mM, PS80 at a concentration of 0.05%, and GSH (or cysteine) at a concentration of 0.4 mM and GSSG (or cystine) at a concentration of 0.2 mM.
• the composition has a pH of 5.5.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH-CDRl consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• VH-CDRl consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17
• sucrose is not part of this composition.
• the disclosure features a pharmaceutical composition comprising an anti-BDCA2 antibody or the BDCA2 -binding fragment thereof at a concentration of 200 mg/ml, sucrose at a concentration of 3%; histidine at a concentration of 20 niM, Arg.HCl at a concentration of 250 mM, and PS80 at a concentration of 0.05%.
• the composition has a pH of 6.0. This pharmaceutical composition is especially suitable for subcutaneous
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH- CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDRl consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• sucrose is not part of this composition.
• the disclosure features a pharmaceutical composition
• a pharmaceutical composition comprising an anti-BDCA2 antibody or the BDCA2-binding fragment thereof at a concentration of 225 mg/ml, sucrose at a concentration of 1%; histidine at a concentration of 20 mM, Arg.HCl at a concentration of 250 mM, and PS80 at a concentration of 0.05%.
• the composition has a pH of 6.0.
• This pharmaceutical composition is especially suitable for subcutaneous administration to a subject in need thereof.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDRl consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• sucrose is not part of this composition.
• the pharmaceutical composition comprises a thiol-containing antioxidant.
• the thiol-containing antioxidant is GSH, GSSG, the combination of GSH and GSSG, cystine, cysteine, or the combination of cysteine and cystine.
• the thiol-containing antioxidant is GSH.
• the thiol-containing antioxidant is GSSG.
• the thiol- containing antioxidant is the combination of GSH and GSSG.
• the thiol- containing antioxidant is cysteine.
• the thiol-containing antioxidant is the combination of cysteine and cystine.
• the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.02 mM to 2 mM. In some instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.2 mM. In other instances, the thiol-containing antioxidant is found in the pharmaceutical composition at a concentration of 0.4 mM. In some instances, the thiol- containing antioxidant is found in the pharmaceutical composition at a concentration of 1.0 mM. In certain cases, GSH and GSSG are found in the pharmaceutical composition at concentrations of 0.4 mM and 0.2 mM, respectively. In other cases, cysteine and cystine are found in the pharmaceutical composition at concentrations of 0.4 mM and 0.2 mM, respectively.
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises a VH and VL, wherein the VH consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:7; and the VL consists of a sequence at least 80% identical, at least 90% identical, or at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:8.
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises an immunoglobulin heavy- chain and an immunoglobulin light chain, wherein the heavy chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:9; and the light chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO: 10.
• the disclosure features a method of treating a condition selected from the group consisting of systemic lupus erythematosus, cutaneous lupus erythematosus, discoid lupus erythematosus, Sjogren's syndrome, dermatopolymyositis, scleroderma, and cytokine release syndrome in a human subject in need thereof.
• the method comprises administering to the human subject a pharmaceutical composition comprising an anti-BDCA2 antibody or BDCA2 -binding fragment described herein.
• the pharmaceutical composition is administered
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof of the pharmaceutical composition is administered to the human subject at a dose of 25 mg every four weeks. In certain embodiments, the anti-BDCA2 antibody or BDC A2-binding fragment thereof of the pharmaceutical composition is administered to the human subject at a dose of 50 mg every four weeks. In certain embodiments, the anti-BDCA2 antibody or BDCA2-binding fragment thereof of the pharmaceutical composition is administered to the human subject at a dose of 150 mg every four weeks. In certain embodiments, the anti-BDCA2 antibody or BDCA2-binding fragment thereof of the pharmaceutical composition is administered to the human subject at a dose of 450 mg every four weeks. In certain instances, the anti-BDCA2 antibody or BDCA2-binding fragment thereof of the pharmaceutical composition is administered to the human subject at the dose corresponding to the human subject's weight as recited below:
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof of the pharmaceutical composition is administered to the human subject at the dose corresponding to the human subject's weight as recited below:
• the disclosure provides a method of treating a condition selected from the group consisting of systemic lupus erythematosus, cutaneous lupus erythematosus, discoid lupus erythematosus, Sjogren's syndrome, dermatopolymyositis, scleroderma, and cytokine release syndrome in a human subject in need thereof.
• the method involves administering subcutaneously to the human subject an anti-BDCA2 antibody or BDCA2- binding fragment thereof at the dose corresponding to the human subject's weight as recited below:
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises a VH and VL, wherein the VH consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:7; and the VL consists of a sequence at least 80% identical, at least 90% identical, or at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:8.
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises an immunoglobulin heavy- chain and an immunoglobulin light chain, wherein the heavy chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:9; and the light chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO: 10.
• human subject is 20 years or less. In certain embodiments, human subject is 18 years or less.
• human subject is 16 years or less. In certain embodiments, human subject is 14 years or less. In certain embodiments, human subject is 12 years or less. In certain embodiments, human subject is 10 years or less. In certain embodiments, human subject is 8 years or less. In certain embodiments, human subject is 6 years or less. In certain embodiments, human subject is 4 years or less. In certain embodiments, human subject is 2 years or less.
• the disclosure features a method of treating a condition selected from the group consisting of systemic lupus erythematosus, cutaneous lupus erythematosus, discoid lupus erythematosus, Sjogren's syndrome, dermatopolymyositis, scleroderma, and cytokine release syndrome in a human subject in need thereof.
• the method involves administering subcutaneously to the human subject an anti-BDCA2 antibody or BDCA2- binding fragment thereof at the dose corresponding to the human subject's weight as recited below:
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDR1 consists of the amino acid sequence set forth in SEQ ID NO:4; VL-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:5; and VL-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:6.
• VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises a VH and VL, wherein the VH consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:7; and the VL consists of a sequence at least 80% identical, at least 90% identical, or at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:8.
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the heavy chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:9; and the light chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO: 10.
• human subject is 20 years or less. In certain embodiments, human subject is 18 years or less.
• human subject is 16 years or less. In certain embodiments, human subject is 14 years or less. In certain embodiments, human subject is 12 years or less. In certain embodiments, human subject is 10 years or less. In certain embodiments, human subject is 8 years or less. In certain embodiments, human subject is 6 years or less. In certain embodiments, human subject is 4 years or less. In certain embodiments, human subject is 2 years or less.
• the disclosure features a syringe or pump comprising a sterile preparation of a pharmaceutical composition described herein adapted for subcutaneous administration of the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a fixed dose of 18 mg, 22 mg, 25 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, 150 mg, or 450 mg.
• the disclosure features a syringe or pump comprising a sterile preparation of a pharmaceutical composition described herein adapted for subcutaneous administration of the anti-BDCA2 antibody or BDCA2-binding fragment thereof at a fixed dose of 18 mg, 22 mg, 25 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, 150 mg, or 450 mg, wherein the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), the VH and VL, respectively, comprising VH complementarity determining regions (CDRs), wherein VH-CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 1 or 17; VH-CDR2 consists of the amino acid sequence set forth in SEQ ID NO:2; and VH-CDR3 consists of the amino acid sequence set forth in SEQ ID NO:3; and VL CDRs, wherein VL-CDR1 consists of
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises a VH and VL, wherein the VH consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:7; and the VL consists of a sequence at least 80% identical, at least 90% identical, or at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:8.
• the anti-BDCA2 antibody or BDCA2 -binding fragment comprises an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the heavy chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:9; and the light chain consists of a sequence at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO: 10.
• FIG. 1 is a graph depicting the viscosity of the antibody formulation.
• FIG. 2A is a graph showing the aggregation of anti-BDCA2 antibodies formulated at a concentration of 150 mg/ml in a formulation containing 20 mM buffer as shown, 140 mM Arg.HCl, and 0.05% PS80 after 0-4 weeks of incubation at 40°C. Buffers are identified by the symbols shown in the figure.
• FIG. 2B is a graph showing the aggregation of anti-BDCA2 antibodies formulated at a concentration of 150 mg/ml in a formulation containing 20 mM buffer as shown, 140 mM Arg.HCl, and 0.05% PS80 after 0-3 months of incubation at 5°C. Buffers are identified using the same symbols as shown in Fig. 2A.
• FIG. 2C is a graph showing the aggregation of anti-BDCA2 antibodies formulated at a concentration of 200 mg/ml in a formulation containing 20 mM buffer as shown, 140 mM Arg.HCl, and 0.05% PS80 after 0-3 months of incubation at 5°C. Buffers are identified using the same symbols as shown in Fig. 2A.
• FIG. 2D is a graph showing the aggregation of anti-BDCA2 antibodies formulated at a concentration of 225 mg/ml in a formulation containing 20 mM buffer as shown, 140 mM Arg.HCl, and 0.05% PS80 after 0-3 months of incubation at 5°C. Buffers are identified using the same symbols as shown in Fig. 2A.
• FIG. 3 is a bar graph depicting the viscosity of anti-BDCA2 antibodies at different pH (5.S, 6, or 6.5), concentration (150 mg/ml, 225 mg/ml, or 250 mg/ml), and in different buffers (citrate or bistidine).
• FIG. 4 is a graph depicting aggregation of BDCA2 at 225 ng/ml in the formulations shown.
• FIG. 5A is a bar graph showing sub-visible particulate formation (particles > 2 um) at time zero (first bar), after 2 weeks at 25°C (second bar) or 2 weeks at 5°C (third bar).
• Particle concentration is depicted on a log scale. Formulations contained the excipient(s) shown, as well as 20 mM Citrate pH 6.0, 0.05% PS80.
• FIG. 5B is a bar graph showing sub-visible particulate formation (particles > 10 um) at time zero (first bar), after 2 weeks at 25°C (second bar) or 2 weeks at 5°C (third bar).
• Particle concentration is depicted on a log scale. Formulations contained the excipient(s) shown, as well as 20 mM Citrate, pH 6.0, and 0.05% PS80.
• FIG. 6 is a bar graph depicting aggregation at time zero (first bar), after 2 weeks at 25°C (second bar) or 2 weeks at 5°C (third bar).
• Formulations contained the excipients shown as well as 20 mM Citrate, pH 6.0, and 0.05% PS80.
• FIG. 7 is a graph comparing aggregation of 150 mg/mL of anti-BDCA2 antibody formulated in Formulation 2 (20 mM His, 100 mM Arg.HCl, 3% sucrose, 0.05% PS80, pH
• FIG. 8 is a graph depicting the viscosity of anti-BDCA2 antibody in Formulation 2.
• FIG. 9 is a graph showing the percentage of high molecular weight species that form over time at 5°C in the ten formulations tested.
• the legend text corresponds to: protein concentration (mg/mL)/Arginine.HCl (mM)/Sucrose (%)/pH.
• FIG. 10 is a graph showing the percentage of high molecular weight species that form overtime at 25°C in the ten formulations tested.
• the legend text corresponds to: protein concentration (mg/mL)/Arginine.HCl (mM)/Sucrose (%)/pH.
• FIG.11 is a graph showing the percentage of high molecular weight species that form overtime at 30°C in the ten formulations tested.
• the legend text corresponds to: protein concentration (mg/mL)/Arginine.HCl (mM)/Sucrose (%)/pH.
• FIG. 12 is a graph showing the percentage of high molecular weight species that form overtime at 40°C in the ten formulations tested.
• the legend text corresponds to: protein concentration (mg/mL)/Arginine.HCl (mM)/Sucrose (%)/pH.
• FIG. 13 is a graph showing the percentage of basic isoforms that form overtime at 25°C in the ten formulations tested.
• the legend text corresponds to: protein concentration (mg/mL)/Arginine.HCl (mM)/Sucrose (%)/pH.
• FIG. 14 is a graph showing the percentage of basic isoforms that form overtime at 30°C in the ten formulations tested.
• the legend text corresponds to: protein concentration (mg/niL)/Arginine.HCl (mM)/Sucrose (%)/pH.
• FIG. 15 is a graph showing the percentage of basic isoforms that form over time at
• FIG. 16 is a graph showing the percentage of basic isoforms that form overtime at 5°C in the ten formulations tested.
• the legend text corresponds to: protein concentration (mg/mL)/Arginine.HCl (mM)/Sucrose (%)/pH.
• FIG. 17 provides graphs depicting the percentage of HMW species of an anti-BDCA2 antibody formulation comprising sucrose (150 mg/ml antibody; 20 mM histidine; 100 mM Arg.HCl; 3% sucrose; 0.05% PS80, pH 5.5) with or without GSH (0.4mM) at 25°C and 40°C.
• FIG. 18 provides an overlay of the graph of Figure 17 with a graph depicting the percentage of HMW species of an anti-BDCA2 antibody formulation lacking sucrose (150 mg/ml antibody; 20 mM histidine; 100 mM Arg.HCl; 0.05% PS80, pH 5.5) with or without GSH (0.4mM) at 25°C and 40°C. This shows that the presence of sucrose has no effect on GSH action.
• FIG. 19 provides graphs depicting the percentage of HMW species of a BENEPALI® (an etanercept biosimilar referencing Enbrel®) formulation (50 mg/ml SB4; 10 mM sodium phosphate; 140 mM NaCl; 1% sucrose, pH 6.2) with or without GSH (0.4mM) at 25°C and 40°C.
• BENEPALI® an etanercept biosimilar referencing Enbrel®
• FIG. 20 provides graphs depicting the percentage of HMW species of an anti-ctvP5 integrin antibody (STX200) formulation (50 mg/ml antibody; 20 mM histidine; 5% sorbitol; 0.05% PS80, pH 6.5) with or without GSH (0.4mM) at 25°C and 40°C.
• STX200 anti-ctvP5 integrin antibody
• This application provides pharmaceutical compositions and dosage regimens of anti- BDCA2 antibodies and BDCA2-binding fragments thereof and their use in the treatment of BDCA2-associated disorders (e.g., SLE, CLE, and DLE).
• BDCA2-associated disorders e.g., SLE, CLE, and DLE.
• BDCA2 is a type ⁇ C-type lectin that is specifically expressed on plasmacytoid dendritic cells (pDCs).
• BDCA2 consists of a single extracellular carbohydrate recognition domain (CRD) at its C-terminus, a transmembrane region, and a short cytoplasmic tail at its N- terminus that does not harbor a signaling motif.
• BDCA2 transmits intracellular signals through an associated transmembrane adaptor, FceRIy.
• Antibody-mediated ligation of BDCA2 leads to recruitment of spleen tyrosine kinase (SYK) to phosphorylated
• ⁇ immunoreceptor tyrosine-based activation motif ( ⁇ ) of FcsRIy.
• Syk activation leads to the activation of B cell linker (Blnk), Bruton's tyrosine kinase (BTK), and phospholipase Cy2 (PLCy2), leading to Ca2 + mobilization.
• Blnk B cell linker
• BTK Bruton's tyrosine kinase
• PLCy2 phospholipase Cy2
• the amino acid sequence of the human BDCA2 protein (Genbank® Accession No. NP 569708.1) is shown below (the transmembrane domain is italicized; the ectodomain is underlined).
• NP_004097.1 is shown below.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof used in the compositions and methods described herein comprises the three heavy chain variable domain complementarity determining regions (CDRs) of an antibody referred to as "BIIB059.”
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises the three light chain variable domain CDRs of ⁇ 059.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises the three heavy chain variable domain CDRs and the three light chain variable domain CDRs of BIIB059.
• the CDRs can be based on any CDR definition in the art, e.g., the definitions of Kabat, Chothia, Chothia from Abysis, enhanced Chothia/AbM, or based on the contact definition.
• CDR sequences of BIIB059 according to these exemplary CDR definitions are provided in Table 1 below.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a VH CDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 1 or 17, a VH CDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO. : 2; and a VH CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO. 3.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a VL CDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.:4, a VL CDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 5; and a VL CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO. 6.
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof comprises the CDRs comprising or consisting of the amino acid sequences set forth in SEQ ID NOs.: 1 to 6. In other embodiments, the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises the CDRs comprising or consisting of the amino acid sequences set forth in SEQ ID NOs.: 11 to 16. In yet other embodiments, the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises the CDRs comprising or consisting of the amino acid sequences set forth in SEQ ID NOs.: 17 to 22.
• the anti- BDCA2 antibody or BDCA2 -binding fragment thereof comprises the CDRs comprising or consisting of the amino acid sequences set forth in SEQ ID NOs.: 23 to 28.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a VH CDRl comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 1 or 17, a VH CDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 2; and a VH CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.
• VL CDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.:4, a VL CDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 5; and a VL CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO. 6.
• B11B059 is an exemplary anti-BDCA2 antibody that can be used in the compositions and methods described herein.
• BIIB059 is a humanized antibody having two glycosylated human IgGl heavy chains and two human kappa light chains that specifically binds to BDCA2 on the surface of plasmacytoid dendritic cells.
• the wild-type IgGl sequence contains a single N-linked glycosylation site and binds to Fc receptors with affinities typical of this class of molecules.
• This Fc function-competent IgGl monoclonal antibody exhibits high affinity for BDCA2 and binds equally well to native human and cynomolgus BDCA2.
• B1IB059 is a potent inhibitor of all TLR9-induced type I interferons (IFNs) as well as other cytokines and chemokines by pDCs. ⁇ 059 is equally potent at inhibiting TLR9-induced type I interferon by pDCs from healthy human donors and SLE patients. BIIB059 specifically inhibits TLR9-induced type I IFN by pDCs and does not impact IFN production by other cell types triggered with different TLR ligand. BIIB059 also causes rapid internalization of BDCA2 from the cell surface.
• IFNs TLR9-induced type I interferons
• BIIB059 specifically inhibits TLR9-induced type I IFN by pDCs and does not impact IFN production by other cell types triggered with different TLR ligand. BIIB059 also causes rapid internalization of BDCA2 from the cell surface.
• BDCA2 Upon stimulation, BDCA2 co-localizes with TLR9 in the endosomal/lysosomal compartment which appears to be necessary for its inhibition of TLR9 signaling. ⁇ 059 was also found to cause CD62L shedding from the surface of human pDCs. In vitro antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) studies suggest that BIIB059 can have cell depletion activity in cell lines overexpressing BDCA2.
• ADCC antibody-dependent cell-mediated cytotoxicity
• CDC complement-dependent cytotoxicity
• variable heavy chain (VH) of ⁇ 059 comprises or consists of the following amino acid sequence:
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a VH having the amino acid sequence set forth in SEQ ID NO:7.
• the anti-BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises a VH domain that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of the VH domain of ⁇ 059 (SEQ ID NO:7), or differs at least at 1 to 5 amino acid residues, but at fewer than 40, 30, 20, 15, or 10, residues, from SEQ ID NO:7.
• these antibodies (i) bind human or cynomolgus monkey BDCA2 but do not significantly bind BDCA2 from phylogenetic species below primates; and/or (ii) inhibit TLR7/TLR9-induced type I interferon and other cytokine or chemokine production by human pDCs; and/or (iii) mediate internalization of BDCA2 from the surface of pDCs; and/or (iv) downregulate CD32a and/or CD62L from the surface of pDCs; and/or (v) deplete pDCs in vitro by ADCC or CDC.
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof comprises a VL having the amino acid sequence set forth in SEQ ID NO:8.
• the anti-BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises a VL domain that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of the VL domain of ⁇ 059 (SEQ ID NO: 8), or differs at least at 1 to 5 amino acid residues, but at fewer than 40, 30, 20, 15, or 10, residues, from SEQ ID NO:8.
• these antibodies (i) bind human or cynomolgus monkey BDCA2 but do not significantly bind BDCA2 from phylogenetic species below primates; and/or (ii) inhibit TLR7/TLR9-induced type I interferon and other cytokine or chemokine production by human pDCs; and/or (iii) mediate internalization of BDCA2 from the surface of pDCs; and/or (iv) downregulate CD32a and/or CD62L from the surface of pDCs; and/or (v) deplete pDCs in vitro by ADCC or CDC.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a VH having the amino acid sequence set forth in SEQ ID NO:7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8.
• the anti-BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises (i) a VH domain that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of the VH domain of BIIB059 (SEQ ID NO:7), and (ii) a VL domain that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of the VL domain of ⁇ 059 (SEQ ID NO:
• these antibodies (i) bind human or cynomolgus monkey BDCA2 but do not significantly bind BDCA2 from phylogenetic species below primates; and/or (ii) inhibit TLR7/TTLR9-induced type I interferon and other cytokine or chemokine production by human pDCs; and/or (iii) mediate internalization of BDCA2 from the surface of pDCs; and/or (iv) downregulate CD32a and/or CD62L from the surface of pDCs; and/or (v) deplete pDCs in vitro by ADCC or CDC.
• VH, VL, HC, and LC sequences CDRs 1, 2, and 3 based on the Kabat definition are both underlined and boldened.
• the italicized and boldened sequence in the VH and HC is the additional N-terminal sequence found in the CDR1 based on enhanced Chothia/AbM definition.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a HC having the amino acid sequence set forth in SEQ ID NO:9.
• the anti-BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises a HC that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of SEQ ID NO:9, or differs at least at 1 to 5 amino acid residues, but at fewer than 40, 30, 20, 15, or 10, residues, from SEQ ID NO:9.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a LC having the amino acid sequence set forth in SEQ ID NO: 10.
• the anti-BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises a LC that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of SEQ ID NO: 10, or differs at least at 1 to 5 amino acid residues, but at fewer than 40, 30, 20, 15, or 10, residues, from SEQ ID NO: 10.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a HC having the amino acid sequence set forth in SEQ ID NO:9 and a LC having the amino acid sequence set forth in SEQ ID NO: 10.
• the anti-BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises (i) a HC that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of SEQ ID NO:9, and (ii) a LC that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of SEQ ID NO: 10; or differs at least at 1 to 5 amino acid residues, but at fewer than 40, 30,
• the anti-BDCA2 antibody is an IgG antibody.
• the anti-BDCA2 antibody has heavy chain constant region chosen from, e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE.
• the anti- BDCA2 antibody is of the IgGl isotype.
• the anti-BDCA2 antibody is of the IgG2 isotype.
• the anti-BDCA2 antibody is of the IgG3 isotype.
• the antibody has a light chain constant region chosen from, e.g., a human kappa or human lambda light chain.
• the anti-BDCA2 antibody is an IgGl/kappa antibody.
• the anti-BDCA2 antibody includes a human Fc region that binds FcyRIIa (CD32a) with an EC 50 of 7 to 15 ug/mL.
• the antibody includes a human Fc region that binds FcyRIIa (CD32a) with an ECso of 10 pg/mL.
• the antibody includes a human Fc region that binds FcyRIIa (CD32a) with an ECso of 11 Mg/mL.
• the antibody includes a human Fc region that binds FcyRIIa (CD32a) with an ECso of 12 pg/mL.
• the heavy chain constant region is human or a modified form of a human constant region.
• the human constant region may include at least 1 and up to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 substitutions.
• the modified human Fc region is a modified human lgGl Fc region.
• the constant region of an anti-BDCA2 antibody may be modified by mutation of one or more amino acid residues to impart a desired functional property (e.g., altered effector function or half-life, reduced glycosylation).
• the N-linked glycosylation site may be substituted to prevent or reduce N-linked glycosylation of Fc region (e.g., human IgGl Fc region).
• the anti-BDCA2 antibody is a full-length (whole) antibody or substantially full-length.
• the protein can include at least one, and preferably two, complete heavy chains, and at least one, and preferably two, complete light chains.
• the anti-BDCA2 antibody is a BDCA2-binding fragment.
• the BDCA2 -binding fragment is a Fab, a Fab ⁇ an F(ab')2, a Facb, an Fv, a single chain Fv (scFv), a sc(Fv)2, or a diabody.
• Antibodies such as ⁇ 059, or BDCA2-binding fragments thereof can be made, for example, by preparing and expressing synthetic genes that encode the recited amino acid sequences or by mutating human germline genes to provide a gene that encodes the recited amino acid sequences. Moreover, this antibody and other anti-BDCA2 antibodies can be produced, e.g., using one or more of the following methods.
• Anti-BDCA2 antibodies or BDCA2-binding fragments may be produced in bacterial or eukaryotic cells. Some antibodies, e.g., Fab's, can be produced in bacterial cells, e.g., E. coli cells. Antibodies can also be produced in eukaryotic cells such as transformed cell lines (e.g., CHO, 293E, COS). In addition, antibodies (e.g., scFv's) can be expressed in a yeast cell such as Pichia (see, e.g., Powers et al., J Immunol Methods. 251:123-35 (2001)), Hanseula, or Saccharomyces.
• a yeast cell such as Pichia (see, e.g., Powers et al., J Immunol Methods. 251:123-35 (2001)), Hanseula, or Saccharomyces.
• a polynucleotide encoding the antibody is constructed, introduced into an expression vector, and then expressed in suitable host cells.
• Polynucleotides encoding an anti-BDCA2 antibody comprising the VH and/or VL, HC and/or LC of the BDCA2 antibodies described herein would be readily envisioned by the ordinarily skilled artisan. Standard molecular biology techniques are used to prepare the recombinant expression vector, transfect the host cells, select for
• transformants culture the host cells and recover the antibody.
• the expression vector should have characteristics that permit amplification of the vector in the bacterial cells. Additionally, when E. coli such as JM109, DH5a, HB101, or XL 1 -Blue is used as a host, the vector must have a promoter, for example, a lacZ promoter (Ward et al., 341:544-546 (1989), araB promoter (Better et al., Science, 240: 1041-1043 (1988)), or T7 promoter that can allow efficient expression in E. coli.
• a promoter for example, a lacZ promoter (Ward et al., 341:544-546 (1989), araB promoter (Better et al., Science, 240: 1041-1043 (1988)
• T7 promoter that can allow efficient expression in E. coli.
• vectors examples include, for example, M13-series vectors, pUC-series vectors, pBR322, pBluescript, pCR-Script, pGEX-5X-l (Pharmacia), "QIAexpress system"
• the expression vector may contain a signal sequence for antibody secretion.
• the pelB signal sequence (Lei et al., J. Bacteriol., 169:4379 (1987)) may be used as the signal sequence for antibody secretion.
• calcium chloride methods or electroporation methods may be used to introduce the expression vector into the bacterial cell.
• the expression vector includes a promoter necessary for expression in these cells, for example, an SV40 promoter (Mulligan et al, Nature, 277: 108 (1979)), MMLV-LTR promoter, EFlapromoter (Mizushimaef al, Nucleic Acids Res., 18:5322 (1990)), or CMV promoter.
• SV40 promoter Mulligan et al, Nature, 277: 108 (1979)
• MMLV-LTR promoter MMLV-LTR promoter
• EFlapromoter Mozushimaef al, Nucleic Acids Res., 18:5322 (1990)
• CMV promoter CMV promoter
• the recombinant expression vectors may carry additional sequences, such as sequences that regulate replication of the vector in host cells (e.g., origins of replication) and selectable marker genes.
• the selectable marker gene facilitates selection of host cells into which the vector has been introduced (see e.g., U.S. Pat. Nos. 4,399,216, 4,634,665 and 5,179,017).
• typically the selectable marker gene confers resistance to drugs, such as G418, hygromycin, or methotrexate, on a host cell into which the vector has been introduced.
• examples of vectors with selectable markers include pMAM, pDR2, pBK-RSV, pBK-CMV, pOPRSV, and pOP13.
• antibodies are produced in mammalian cells.
• Exemplary mammalian host cells for expressing an antibody include Chinese Hamster Ovary (CHO cells) (including dhfr ⁇ CHO cells, described in Urlaub and Chasin (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, used with a DHFR selectable marker, e.g., as described in Kaufman and Sharp (1982) Mol. Biol.
• Chinese Hamster Ovary CHO cells
• dhfr ⁇ CHO cells described in Urlaub and Chasin (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, used with a DHFR selectable marker, e.g., as described in Kaufman and Sharp (1982) Mol. Biol.
• human embryonic kidney 293 cells e.g., 293, 293E, 293T
• COS cells e.g., N1H3T3 cells
• lymphocytic cell lines e.g., NSO myeloma cells and SP2 cells
• a cell from a transgenic animal e.g., a transgenic mammal.
• the cell is a mammary epithelial cell.
• a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain of an anti-BDCA2 antibody (e.g., ⁇ 059) is introduced into dhfr CHO cells by calcium phosphate-mediated transfection.
• the antibody heavy and light chain genes are each operatively linked to enhancer/promoter regulatory elements (e.g., derived from SV40, CMV, adenovirus and the like, such as a CMV enhancer/AdMLP promoter regulatory element or an SV40 enhancer/AdMLP promoter regulatory element) to drive high levels of transcription of the genes.
• enhancer/promoter regulatory elements e.g., derived from SV40, CMV, adenovirus and the like, such as a CMV enhancer/AdMLP promoter regulatory element or an SV40 enhancer/AdMLP promoter regulatory element
• the recombinant expression vector also carries a DHFR gene, which allows for selection of CHO cells that have been transfected with the vector using methotrexate selection/amplification.
• the selected transformant host cells are cultured to allow for expression of the antibody heavy- and light chains and the antibody is recovered from the culture medium.
• Antibodies can also be produced by a transgenic animal.
• U.S. Pat. No. 5,849,992 describes a method of expressing an antibody in the mammary gland of a transgenic mammal.
• a transgene is constructed that includes a milk-specific promoter and nucleic acids encoding the antibody of interest and a signal sequence for secretion.
• the milk produced by females of such transgenic mammals includes, secreted-therein, the antibody of interest.
• the antibody can be purified from the milk, or for some applications, used directly. Animals are also provided comprising one or more of the nucleic acids described herein.
• the antibodies of the present disclosure can be isolated from inside or outside (such as medium) of the host cell and purified as substantially pure and homogenous antibodies. Methods for isolation and purification commonly used for antibody purification may be used for the isolation and purification of antibodies, and are not limited to any particular method.
• Antibodies may be isolated and purified by appropriately selecting and combining, for example, column chromatography, filtration, ultrafiltration, salting out, solvent precipitation, solvent extraction, distillation, immunoprecipitation, SDS-polyacrylamide gel electrophoresis, isoelectric focusing, dialysis, and rectystallization. Chromatography includes, for example, affinity chromatography, ion exchange chromatography, hydrophobic chromatography, gel filtration, reverse-phase chromatography, and adsorption
• Chromatography can be carried out using liquid phase chromatography such as HPLC and FPLC.
• Columns used for affinity chromatography include protein A column and protein G column. Examples of columns using protein A column include Hyper D, POROS, and Sepharose FF (GE Healthcare Biosciences).
• the present disclosure also includes antibodies that arc highly purified using these purification methods.
• compositions comprising the anti-BDCA2 antibodies or BDCA2-binding fragments thereof described herein.
• the anti-BDCA2 antibody compositions comprises an anti-BDCA2 antibody or BDCA2-binding fragment thereof comprising an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), wherein the VH comprises the H-CDRs and the VL comprises the L-CDRs of ⁇ 059.
• VH immunoglobulin heavy chain variable domain
• VL immunoglobulin light chain variable domain
• the H-CDRs of comprise or consist of the amino acid sequences set forth in SEQ ID NO: 1 or 17, SEQ ID NO:2, and SEQ ID NO:3; and the L-CDRs comprise or consist of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6.
• the anti-BDCA2 antibody compositions comprises an anti-BDCA2 antibody or BDCA2 -binding fragment thereof comprising (i) a VH comprising or consisting of an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ YD NO: 7; and (ii) a VL comprising or consisting of an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO:8.
• the anti-BDCA2 antibody compositions comprises an anti-BDCA2 antibody comprising (i) a heavy chain comprising or consisting of an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO:9; and (ii) a light chain comprising or consisting of an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.
• these compositions are high concentration anti-BDCA2 antibody composition.
• high concentration anti-BDCA2 antibody composition is meant a composition comprising anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of greater than 50 mg/ml and less than 300 mg/ml.
• the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of SO mg/ml to 240 mg/ml.
• the anti- BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 50 mg/ml to 225 mg/ml.
• the anti- BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2 -binding fragments thereof at a concentration of 75 mg/ml to 225 mg/ml. In other instances, the anti- BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 100 mg/ml to 225 mg/ml. In yet other instances, the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2 -binding fragments thereof at a concentration of 125 mg/ml to 225 mg/ml.
• the anti- BDC A2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 125 mg/ml to 175 mg/ml. In certain instances, the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 240 mg/ml. In certain instances, the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2 -binding fragments thereof at a concentration of 225 mg/ml. In certain instances, the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 200 mg/ml.
• the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 175 mg/ml. In certain instances, the anti-BDCA2 antibody composition comprises anti- BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 150 mg/ml. In other instances, the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2 -binding fragments thereof at a concentration of 125 mg/ml. In some instances, the anti-BDCA2 antibody composition comprises anti-BDCA2 antibodies or BDCA2-binding fragments thereof at a concentration of 100 mg/ml.
• a composition comprising an anti-BDCA2 antibody or BDCA2-binding fragment thereof described herein may be in any one of a variety of forms. These include, for example, liquid solutions (e.g., injectable and infusible solutions), dispersions, or suspensions. The preferred form can depend on the intended mode of administration and therapeutic application.
• a pharmaceutical composition described herein is in the form of a sterile injectable or infusible solution.
• Sterile injectable solutions can be prepared by incorporating an antibody described herein in the required amount with one or a combination of ingredients, followed by filtered sterilization.
• dispersions are prepared by incorporating an antibody described herein into a sterile vehicle that contains a basic dispersion medium and the required other ingredients.
• an exemplary method of preparation is vacuum drying and freeze drying that yields a powder of an antibody described herein plus any additional desired ingredient from a previously sterile-filtered solution thereof.
• the proper fluidity of a solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants.
• the anti-BDCA2 antibody compositions may additionally comprise one or more excipients.
• the excipient lowers/reduces the aggregation and/or viscosity of the antibody in the composition compared to aggregation and/or viscosity of the antibody in the pharmaceutical composition without that excipient.
• such an excipient is arginine.
• the excipient is arginine hydrochloride.
• Arginine e.g., arginine hydrochloride
• arginine hydrochloride can be included in the composition at a concentration of 50 mM to 250 mM, 50 mM to 200 mM, 50 mM to 150 mM, 50 mM to 125 mM, 50 mM to 100 mM, 75 mM to 250 mM, 75 mM to 200 mM, 75 mM to 150 mM, or 75 mM to 100 mM.
• arginine e.g., Arg.HCl
• arginine e.g., Arg.HCl
• arginine is present in the composition at a concentration of 50 mM to 200 mM.
• arginine e.g., arginine hydrochloride
• arginine hydrochloride can be included in the composition at a concentration of 100 mM, 120 mM, 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, or 150 mM.
• arginine e.g., arginine hydrochloride
• arginine e.g., arginine hydrochloride
• arginine e.g., arginine hydrochloride
• solutions containing arginine develop visible particles after incubation at room temperature or higher temperatures (e.g., 40°C).
• room temperature or higher temperatures e.g. 40°C.
• sucrose can reduce or prevent the formation of visible particles.
• sucrose was also unexpectedly found to lower the counts of subvisible particulates.
• the anti-BDCA2 antibody composition comprises sucrose at a concentration of 0.05% to 15%, 0.05% to 10%, 0.05% to 5%, l% to 15 %, l%to 10%, l% to 5%, 2% to 8%, 2% to 6%, or 2% to 4%.
• the anti-BDCA2 antibody composition comprises sucrose at a concentration of 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5% 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10%.
• the anti-BDCA2 antibody composition comprises sucrose at a concentration of 3%.
• the anti-BDCA2 antibody composition comprises sucrose at a concentration of 1%.
• Antibody product manufacturing is a complex process that can involve several steps such as, e.g., drug substance and bulk formulation, filtration, shipping, pooling, filling, lyophilization, inspections, packaging, and storage. During these steps, antibodies may be subjected to many different forms of stresses, e.g., agitation, temperature, light exposure, and oxidation. These types of stresses can lead to denaturation and aggregation of the antibody, which compromise the product quality and can even lead to loss of a production batch.
• the composition may include a polysorbate.
• the composition comprises polysorbate-80 at a concentration of 0.01% to 0.5%, 0.01% to 0.1%, 0.01% to 0.09%, 0.01% to 0.08%, 0.01% to 0.07%, 0.01% to 0.06%, 0.01% to 0.05%, 0.01% to 0.04%, or 0.01% to 0.03%.
• the composition comprises polysorbate-80 at a concentration of 0.02% to 0.08%. In some embodiments, the composition comprises polysorbate-80 at a concentration of 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.1%. In a particular embodiment, the composition comprises polysorbate-80 at a concentration of 0.05%.
• the antibody composition comprises histidine as the buffering agent.
• the composition comprises histidine at a concentration of 5 raM to 50 mM, 5 mM to 40 mM, 5 mM to 30 mM, 5 raM to 25 mM, 10 mMto SO mM, 10 mM to 40 mM, 10 mMto 30 mM, 10 mM to 25 mM, 15 mM to 50 mM, 15 mM to 40 mM, 15 mM to 30 mM, or 15 mM to 25 mM.
• the composition comprises histidine at a concentration of 10 mM to 30 mM.
• the composition comprises histidine at a concentration of 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, or 30 mM. In a particular embodiment, the composition comprises histidine at a concentration of 20 mM.
• the pH of the antibody composition can be 5.0 to 6.5. In certain cases, the pH of the antibody composition can be 5.0 to 6.0. In certain instances, the pH of the antibody composition is 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, or 6.5. In a particular embodiment, the pH of the antibody composition is 5.5. In another particular embodiment, the pH of the antibody composition is 6.0. In yet another particular embodiment, the pH of the antibody composition is 6.5.
• the composition comprises a thiol-containing antioxidant (e.g., reduced glutathione (GSH), oxidized glutathione (GSSG), GSH + GSSG, cysteine, cystine, cysteine + cystine) at a concentration of 0.02 mM to 2 oiM (e.g., 0.02, 0.03, 0.05, 0.06, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0 mM).
• a thiol-containing antioxidant e.g., reduced glutathione (GSH), oxidized glutathione (GSSG), GSH + GSSG, cysteine, cystine, cysteine + cystine
• 0.02 mM to 2 oiM e.g., 0.02, 0.03, 0.05, 0.06, 0.08, 0.1, 0.2
• the composition comprises GSH at a concentration of 0.4 mM. In some cases, the composition comprises GSSG at a concentration of 0.2 mM. In some cases, the composition comprises GSH at a concentration of 0.4 mM and GSSG at a concentration of 0.2 mM. In some cases, the composition comprises cysteine at a concentration of 0.4 mM.
• the composition comprises cystine at a concentration of 0.2 mM. In some cases, the composition comprises cysteine at a concentration of 0.4 mM and cystine at a concentration of 0.2 mM. In certain embodiments, the composition comprises methionine at a concentration of 5 mM to 15 mM (e.g., 10 mM). In certain embodiments, the composition comprises glutamic acid at a concentration of 50 mM to 80 mM (e.g., 70 mM).
• the composition (e.g., a pharmaceutical composition) comprises an anti-BDCA2 antibody or a BDCA2 -binding fragment thereof at a concentration of 50 mg/ml to 225 mg/ml, sucrose at a concentration of 0.05% to 10%, arginine (e.g., arginine hydrochloride) at a concentration of 50 mM to 250 mM, polysorbate-80 at a concentration of 0.01% to 0.1%, and histidine at a concentration of 10 mM to 30 mM.
• the composition has a pH of 5.0 to 6.0.
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof of the composition comprises a VH and a VL comprising the CDRs of ⁇ 059 (e.g.. SEQ ID NOs.: 1 or 17, 2, 3, 4, 5, and 6).
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof of the composition comprises a VH and a VL comprising SEQ ID NOs: 7 and 8, respectively.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof of the composition comprises a heavy chain and a light chain comprising SEQ ID NOs: 9 and 10, respectively.
• the composition has a pH of 5.5 and comprises BIIB059 or a BIIB059-binding fragment thereof at a concentration of 150 mg/ml, sucrose at a concentration of 3%, arginine hydrochloride at a concentration of 100 mM, polysorbate-80 at a concentration of 0.05%, and histidine at a concentration of 20 mM.
• composition further comprises a thiol-containing antioxidant (e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, cysteine + cystine) at a concentration of 0.02 mM to 2 mM.
• a thiol-containing antioxidant e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, cysteine + cystine
• the composition (e.g., a pharmaceutical composition) comprises an anti-BDCA2 antibody or a BDCA2-binding fragment thereof, arginine (e.g., arginine hydrochloride) at a concentration of 50 mM to 250 mM, polysorbate-80 at a concentration of 0.02% to 0.08%, and histidine at a concentration of 10 mM to 30 mM.
• the composition has a pH of 5.0 to 6.5.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof is present in the composition at a concentration of 50 mg/ml to 225 mg/ml.
• the anti-BDCA2 antibody or BDCA2 -binding fragment thereof of the composition comprises a VH and a VL comprising the CDRs of ⁇ 059 (e.g., SEQ ID NOs.: 1 or 17, 2, 3, 4, 5, and 6).
• the anti- BDCA2 antibody or BDCA2-binding fragment thereof of the composition comprises a VH and a VL comprising SEQ ID NOs: 7 and 8, respectively.
• the anti- BDCA2 antibody or BDCA2-binding fragment thereof of the composition comprises a heavy chain and a light chain comprising SEQ ID NOs: 9 and 10, respectively.
• the composition comprises sucrose at a concentration of 1% to 10%.
• the composition comprises a thiol-containing antioxidant (e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, or cysteine + cystine) at a concentration of 0.02 mM to 2 mM.
• a thiol-containing antioxidant e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, or cysteine + cystine
• the composition has a pH of 5.5 and comprises BIIB059 or a BIIB059-binding fragment thereof at a concentration of 150 mg/ml, sucrose at a
• the above-listed composition further comprises a thiol-containing antioxidant (e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, or cysteine + cystine) at a concentration of 0.02 mM to 2 mM.
• the thiol-containing antioxidant is GSH at a concentration of 0.4 mM.
• the composition may comprise higher concentration of the anti-BDCA2 antibody or BDCA2- binding fragment thereof.
• a composition comprises an anti-BDCA2 antibody or a BDCA2-binding fragment thereof at a concentration of 200 mg/ml; arginine (e.g., arginine hydrochloride) at a concentration of 250 mM; sucrose at a concentration of 3%; polysorbate-80 at a concentration of 0.05%; and histidine at a concentration of 20 mM.
• the pH of this composition is 6.0.
• the composition further comprises a thiol-containing antioxidant (e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, or cysteine + cystine) at a concentration of 0.02 mM to 2 mM.
• a thiol-containing antioxidant e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, or cysteine + cystine
• the thiol-containing antioxidant is GSH at a concentration of 0.4 mM.
• the thiol-containing antioxidant is GSSG at a concentration of 0.2 mM.
• the thiol-containing antioxidant is GSH at a concentration of 0.4 mM and GSSG at a concentration of 0.2 mM.
• such a high concentration composition comprises an anti-BDCA2 antibody or a BDCA2-binding fragment thereof at a concentration of 225 mg/ml; arginine (e.g., arginine hydrochloride) at a concentration of 250 mM; sucrose at a concentration of 1%; polysorbate-80 at a
• the composition further comprises a thiol-containing antioxidant (e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, or cysteine + cystine) at a concentration of 0.02 mM to 2 mM.
• a thiol-containing antioxidant e.g., GSH, GSSG, GSH + GSSG, cysteine, cystine, or cysteine + cystine
• the thiol-containing antioxidant is GSH at a concentration of 0.4 mM.
• the thiol-containing antioxidant is GSSG at a concentration of 0.2 mM.
• the thiol- containing antioxidant is GSH at a concentration of 0.4 mM and GSSG at a concentration of 0.2 mM. In another specific instance, the thiol-containing antioxidant is cysteine at a concentration of 0.4 mM.
• the anti-BDCA2 antibody or BDCA2- binding fragment thereof of the composition comprises a VH and a VL comprising the CDRs of ⁇ 059 (e.g., SEQ ID NOs.: 1 or 17, 2, 3, 4, 5, and 6).
• the anti- BDCA2 antibody or BDCA2-binding fragment thereof of the composition comprises a VH and a VL comprising SEQ ID NOs: 7 and 8, respectively.
• the anti- BDCA2 antibody or BDCA2-binding fragment thereof of the composition comprises a heavy chain and a light chain comprising SEQ ID NOs: 9 and 10, respectively.
• the anti-BDCA2 antibody (e.g., ⁇ 059) or BDCA2-binding fragment thereof described above can be administered to a subject, e.g., a human subject, at different doses.
• the anti-BDCA2 antibody (e.g., ⁇ 059) or BDCA2-binding fragment thereof can be administered as a fixed dose (; ' . e., independent of the weight of the patient), or in a mg/kg dose (i.e., a dose which varies based on the weight of the subject).
• Dosage unit form or "fixed dose” as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier and optionally in association with the other agent. Single or multiple dosages may be given. The treatment can continue for days, weeks, months or even years.
• the dosage of the anti- BDCA2 antibody (e.g., BIIB059) or BDCA2-binding fragment thereof is a fixed dose of 25 mg.
• the dosage of the anti- BDC A2 antibody or BDCA2-binding fragment thereof is a fixed dose of 50 mg.
• the dosage of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof is a fixed dose of 150 mg.
• the dosage of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is a fixed dose of 450 mg.
• the dosage of the anti- BDCA2 antibody (e.g., BIIB059) or BDCA2-binding fragment thereof is a fixed dose of 18 mg, where the child has a weight of 10 to 18 kg.
• the dosage of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is a fixed dose of 22 mg, where the child has a weight of 18.1 kg to 25 kg.
• the dosage of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is a fixed dose of 28 mg, where the child has a weight of 25.1 kg to 48 kg.
• the dosage of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is a fixed dose of 50 mg, where the child has a weight of greater than 48 kg.
• the dosage of the anti- BDCA2 antibody (e.g., ⁇ 059) or BDCA2-binding fragment thereof is a fixed dose of 40 mg, where the child has a weight of 10 to 18 kg.
• the dosage of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is a fixed dose of 56 mg, where the child has a weight of 18.1 kg to 25 kg.
• the dosage of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is a fixed dose of 80 mg, where the child has a weight of 25.1 kg to 48 kg.
• the dosage of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is a fixed dose of 1 SO mg, where the child has a weight of greater than 48 kg.
• the fixed doses described above may each be administered daily, every week, every 2 weeks, every 4 weeks, every 6 weeks, every 8 weeks, monthly, biweekly, weekly, or daily, as appropriate, over a period of time to encompass at least 2 doses, 3 doses, 4 doses, 5 doses, 6 doses, 7 doses, 8 doses, 9 doses, 10 doses, 12 doses, 14 doses, 16 doses, 18 doses, 20 doses, 22 doses, 24 doses or more.
• BDCA2-binding fragment thereof is administered to a human subject every 2 weeks or every 4 weeks for a period of time determined to be beneficial for the subject by her/his healthcare provider.
• a fixed dose of 25 mg of the anti-BDCA2 antibody or BDCA2- binding fragment thereof is administered to a human subject every 4 weeks.
• the subject is also administered a loading dose of 25 mg, 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof two weeks after the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is administered to the subject.
• the loading dose is 25 mg of the anti- BDCA2 antibody or BDCA2-binding fragment thereof.
• the loading dose is 50 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof.
• the subject is administered at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 doses of the fixed dose of 25 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered 4, 5, 6, 7, 8, 9, or 10 doses of the fixed dose of 25 mg of the anti-BDCA2 antibody or BDCA2- binding fragment thereof.
• the subject is administered 2 to 24, 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, or 2 to 8 doses of the fixed dose of 25 mg of the anti- BDCA2 antibody or BDCA2 -binding fragment thereof.
• a fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof is administered to a human subject every 2 weeks or every 4 weeks for a period of time determined to be beneficial for the subject by her/his healthcare provider. In some instances, a fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2- binding fragment thereof is administered to a human subject every 4 weeks. In certain embodiments, the subject is also administered a loading dose of 25 mg, 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof two weeks after the first dose of the anti-BDC A2 antibody or BDCA2-binding fragment thereof is administered to the subject.
• the loading dose is 50 mg of the anti- BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 doses of the fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof.
• the subject is administered 4, 5, 6, 7, 8, 9, or 10 doses of the fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered 2 to 24, 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, or 2 to 8 doses of the fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2- binding fragment thereof.
• a fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is administered to a human subject every 2 weeks or every 4 weeks for a period of time determined to be beneficial for the subject by her/his healthcare provider. In some instances, a fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2- binding fragment thereof is administered to a human subject every 4 weeks. In certain embodiments, the subject is also administered a loading dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof two weeks after the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is administered to the subject.
• the loading dose is 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 doses of the fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered 4, 5, 6, 7, 8, 9, or 10 doses of the fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered 2 to 24, 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, or 2 to 8 doses of the fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof.
• a fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof is administered to a human subject every 2 weeks or every 4 weeks for a period of time determined to be beneficial for the subject by her/his healthcare provider. In some instances, a fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2- binding fragment thereof is administered to a human subject every 4 weeks. In certain embodiments, the subject is also administered a loading dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof two weeks after the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is administered to the subject.
• the loading dose is 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 doses of the fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof.
• the subject is administered 4, 5, 6, 7, 8, 9, or 10 doses of the fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is administered 2 to 24, 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, or 2 to 8 doses of the fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• a pharmaceutical composition may include a "therapeutically effective amount" of an agent described herein. Such effective amounts can be determined based on the effect of the administered agent, or the combinatorial effect of agents if more than one agent is used. A therapeutically effective amount of an agent may also vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the compound to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic, or detrimental effects, of the composition is outweighed by the therapeutically beneficial effects. In one embodiment, the therapeutically effective amount of the anti- BDCA2 antibody or BDCA2 -binding fragment thereof is 25 mg.
• the therapeutically effective amount of the anti-BDCA2 antibody or BDCA2-binding fragment thereof is 50 mg. In another embodiment, the therapeutically effective amount of the anti- BDCA2 antibody or BDCA2-binding fragment thereof is 150 mg. In yet another embodiment, the therapeutically effective amount of the anti-BDCA2 antibody or BDCA2- binding fragment thereof is 450 mg. In one embodiment, the therapeutically effective amount of the anti-BDCA2 antibody or BDC A2-binding fragment thereof for a pediatric human subject (e.g., a subject 21 years of age or less, a subject 18 years of age or less, or a subject 16 years of age or less) is 18 mg, 22 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, or 150 mg.
• a pediatric human subject e.g., a subject 21 years of age or less, a subject 18 years of age or less, or a subject 16 years of age or less
• a pediatric human subject e.g., a subject 21 years of age or less, a subject 18 years
• the anti-BDCA2 antibody or BDCA2-binding compositions described above are administered to the subject at a dose of 25 mg. In other instances, the anti-BDCA2 antibody or BDCA2-binding compositions described above are administered to the subject at a dose of 50 mg. In yet other instances, the anti-BDCA2 antibody or BDCA2- binding compositions described above are administered to the subject at a dose of 150 mg. In certain instances, the anti-BDCA2 antibody or BDCA2-binding compositions described above are administered to the subject at a dose of 450 mg.
• the dose is determined based on the weight of the child as follows:
• the route and/or mode of administration of the anti-BDCA2 antibody or BDCA2- binding fragment thereof can be tailored for the individual subject.
• the route of administration is one of: subcutaneous injection (SC), intravenous injection or infusion (IV), intraperitoneal administration (IP), or intramuscular injection.
• SC subcutaneous injection
• IV intravenous injection or infusion
• IP intraperitoneal administration
• intramuscular injection intramuscular injection.
• the route of administration is subcutaneous.
• the route of administration is intravenous.
• compositions that comprise the anti-BDCA2 antibody or BDCA2- binding fragment thereof alone or in combination with non-BDCA2 antibody agent(s) can be administered with a medical device.
• the device can be designed with features such as portability, room temperature storage, and ease of use so that it can be used in emergency situations, e.g., by an untrained subject or by emergency personnel in the field, removed to medical facilities and other medical equipment.
• the device can include, e.g., one or more housings for storing pharmaceutical preparations that include the anti-BDCA2 antibody or BDCA2-binding fragment thereof, and can be configured to deliver one or more unit doses of the blocking agent.
• the pharmaceutical composition can be administered with a needleless hypodermic injection device, such as the devices disclosed in US 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; or 4,596,556.
• a needleless hypodermic injection device such as the devices disclosed in US 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; or 4,596,556.
• implants and modules examples include: US 4,487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; US 4,486, 194, which discloses a therapeutic device for administering medicaments through the skin; US 4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; US 4,447,224, which discloses a variable flow implantable infusion apparatus for continuous drug delivery; US 4,439,196, which discloses an osmotic drug delivery system having multi- chamber compartments; and US 4,475,196, which discloses an osmotic drug delivery system. Many other devices, implants, delivery systems, and modules are also known.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof is administered to a human subject with a syringe. In another embodiment, the anti-BDCA2 antibody or BDCA2-binding fragment thereof is administered to a human subject with a pump for subcutaneous delivery. In some embodiments, the anti-BDC A2 antibody or
• BDCA2 -binding fragment thereof is administered to a human subject with an autoinjector.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof is administered to a human subject with a subcutaneous large volume injector.
• This disclosure provides a pump or syringe comprising a sterile preparation of an anti- BDCA2 antibody (e.g., BIIB059) or BDCA2-binding fragment thereof.
• the syringe or pump can be adapted for subcutaneous administration of the anti-BDCA2 antibody or BDCA2- binding fragment thereof.
• the syringe or pump delivers a fixed doses(s) (e.g., 18 mg, 22 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, 150 mg, 450 mg) of the anti-BDCA2 antibody or BDC A2-binding fragment thereof.
• the disclosure also provides a pump, syringe, or injector (e.g., autoinjector, subcutaneous large volume injector) comprising a sterile preparation of the pharmaceutical compositions described above.
• the syringe or pump can be adapted for subcutaneous administration of the pharmaceutical compositions comprising the anti-BDCA2 antibody or BDC A2 -binding fragment thereof.
• the syringe or pump delivers a fixed doses(s) (e.g., 18 mg, 22 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, 150 mg, 450 mg) of the anti-BDCA2 antibody or BDCA2 -binding fragment thereof.
• An anti-BDCA2 antibody or BDCA2 -binding fragment thereof described herein can be used to treat or prevent a variety of immunological disorders, such as inflammatory and autoimmune disorders.
• Anti-BDCA2 antibodies or BDCA2-binding fragments thereof can disable or deplete pDCs, and/or inhibit inflammatory cytokines and chemokines produced by pDCs, and/or downregulate CD32a, and/or inhibiting immune complex stimulation of pDCs, and/or downregulate or cause shedding of CD62L.
• the anti-BDCA2 antibodies or BDCA2- binding fragment thereof of this disclosure can be combined with an antimalarial agent (e.g., HCQ) for improved therapeutic effects in the treatment of inflammatory and autoimmune disorders.
• an antimalarial agent e.g., HCQ
• Anti-BDCA2 antibodies can be used to reduce levels of cytokines and chemokines such as: type I interferons, type III interferons, IL-6, TNF-o, MlPl-a and MIPl- ⁇ , CCL5, and IP-10.
• Type I IFNs constitute a multiple-member family of cytokines, including 13 IFN-a subtypes, IFN- ⁇ , - ⁇ , - ⁇ , - ⁇ , - ⁇ and - ⁇ .
• Type III interferons consist of three IFN- ⁇ molecules called IFN- ⁇ , IFN-X2 and IFN ⁇ 3 (also referred to as IL29, IL28A and IL28B, respectively).
• IFN- ⁇ IFN- ⁇ molecules
• IFN-X2 IFN-X2
• IFN ⁇ 3 also referred to as IL29, IL28A and IL28B, respectively.
• the anti-BDCA2 antibodies described herein provide a more robust treatment approach than treatments attempting to reduce specific IFN subtypes with neutralizing antibodies.
• the pDC-focused treatment approach of the anti-BDCA2 antibodies is more selective and potentially safer than global blockade of the IFN response.
• anti-BDCA2 antibodies described herein effectively eliminate pDC-derived type I IFNs while maintaining other sources of IFN that could be necessary in the event of viral infections.
• BDCA2 -associated disorders include SLE, CLE, DLE, lupus nephritis, systemic sclerosis (scleroderma), morphea, psoriasis, rheumatoid arthritis, inflammatory bowel disease (IBD),
• the anti-BDCA2 antibodies and compositions described herein can be used to treat a lupus disorder (e.g., SLE, CLE, and DLE).
• a lupus disorder e.g., SLE, CLE, and DLE.
• the disclosure features a method of treating SLE (e.g., moderate or severe lupus) in a human subject in need thereof.
• the method involves administering to a human subject in need thereof a therapeutically effective amount of an anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject is administered the pharmaceutical compositions described herein to provide a dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject when the subject is a pediatric subject (e.g., a subject 21 years of age or less, a subject 18 years of age or less, or a subject 16 years of age or less), the subject is administered the pharmaceutical compositions described herein to provide a dose of 18 mg, 22 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, or 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the dose is chosen based on the weight of the child as detailed above.
• the subject is administered at least 2, at least 3, at least 4, at least 5, at least 6, at least 7 at least 8, at least 9, at least 10 doses, at least 11 doses, at least 12 doses, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, of 12 doses.
• the subject is also administered a loading dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment about 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2 -binding fragment.
• the subject with SLE is administered a fixed dose of 50 mg of the anti- BDCA2 antibody or BDCA2-binding fragment and a loading dose of 50 mg of the anti- BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDC A2-binding fragment.
• the subject with SLE is administered a fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti- BDCA2 antibody or BDCA2-binding fragment.
• the subject with SLE is administered a fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2 -binding fragment.
• the disclosure also features a method of treating cutaneous lupus erythematosus (with or without SLE) in a human subject in need thereof.
• the method involves administering to a human subject in need thereof a therapeutically effective amount of an anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject is administered the pharmaceutical compositions described herein to provide a dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject when the subject is a pediatric subject (e.g., a subject 21 years of age or less, a subject 18 years of age or less, or a subject 16 years of age or less), the subject is administered the pharmaceutical compositions described herein to provide a dose of 18 mg, 22 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, or 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the dose is chosen based on the weight of the child as detailed above.
• the subject is administered at least 2, at least 3, at least 4, at least 5, at least 6, at least 7 at least 8, at least 9, at least 10 doses, at least 11 doses, at least 12 doses, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, of 12 doses.
• the subject is also administered a loading dose of SO mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment about 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with CLE (with or without SLE) is administered a fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 50 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2 -binding fragment.
• the subject with CLE (with or without SLE) is administered a fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with CLE (with or without SLE) is administered a fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the disclosure also provides a method of treating discoid lupus erythematosus (with or without SLE) in a human subject in need thereof.
• the method involves administering to a human subject in need thereof a therapeutically effective amount of an anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject is administered the pharmaceutical compositions described herein to provide a dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject when the subject is a pediatric subject (e.g., a subject 21 years of age or less, a subject 18 years of age or less, or a subject 16 years of age or less), the subject is administered the pharmaceutical compositions described herein to provide a dose of 18 mg, 22 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, or 150 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment.
• the dose is chosen based on the weight of the child as detailed above.
• the subject is administered at least 2, at least 3, at least 4, at least 5, at least 6, at least 7 at least 8, at least 9, at least 10 doses, at least 11 doses, at least 12 doses, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, of 12 doses.
• the subject is also administered a loading dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment about 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with discoid lupus (with or without SLE) is administered a fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 50 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with discoid lupus (with or without SLE) is administered a fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with discoid lupus (with or without SLE) is administered a fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the disclosure features a method of treating cytokine release syndrome and/or cytokine storms in a human subject in need thereof.
• Cytokine release syndrome is a common immediate complication occurring with the use of T cell- engaging therapies (e.g., chimeric antigen receptor-modified T cell (CART) therapy). Severe cases of this disorder are known as cytokine storms.
• CRS is a symptom complex associated with the use of many monoclonal antibodies. Commonly referred to as an infusion reaction, CRS results from the release of cytokines from cells targeted by the antibody as well as immune effector cells recruited to the area. The antibodies bind to the T cell receptor, activating the T cells before they are destroyed.
• the cytokines released by the activated T cells produce a type of systemic inflammatory response similar to that found in severe infection.
• the subject can develop systemic symptoms such as fever, nausea, chills, hypotension, tachycardia, asthenia, headache, rash, scratchy throat, and dyspnea.
• the symptoms are mild to moderate in severity and can be managed relatively easily.
• some patients can experience severe, life- threatening reactions that result from massive release of cytokines. Severe reactions occur more commonly during the first infusion in patients with hematologic malignancies who have not received prior chemotherapy. Severe reactions are marked by their rapid onset and the acuity of associated symptoms.
• the method of treating CRS involves administering to a human subject in need thereof an anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject is administered the pharmaceutical compositions described herein to provide a dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2 -binding fragment.
• the subject when the subject is a pediatric subject (e.g., a subject 21 years of age or less, a subject 18 years of age or less, or a subject 16 years of age or less), the subject is administered the pharmaceutical compositions described herein to provide a dose of 18 mg, 22 mg, 28 mg, 40 mg, 50 mg, 56 mg, 80 mg, or 150 mg of the anti- BDCA2 antibody or BDCA2-binding fragment.
• the dose is chosen based on the weight of the child as detailed above.
• the subject is administered at least 2, at least 3, at least 4, at least 5, at least 6, at least 7 at least 8, at least 9, at least 10 doses, at least 11 doses, at least 12 doses, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, of 12 doses.
• the subject is also administered a loading dose of 50 mg, 150 mg, or 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment about 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with CRS is administered a fixed dose of 50 mg of the anti-BDCA2 antibody or BDCA2- binding fragment and a loading dose of 50 mg of the anti-BDCA2 antibody or BDCA2- binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with CRS is administered a fixed dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 150 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the subject with CRS is administered a fixed dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment and a loading dose of 450 mg of the anti-BDCA2 antibody or BDCA2-binding fragment at 2 weeks after the administration of the first dose of the anti-BDCA2 antibody or BDCA2-binding fragment.
• the human subject has, is scheduled to, or is undergoing CART therapy (e.g., CART-19 therapy). In certain instances, the human subject has, is scheduled to, or is undergoing therapy with an anti-T cell antibody (e.g., ATG, OKT3,
• the subject has, is scheduled to, or is undergoing therapy with an anti-CD20 antibody (e.g., rituximab).
• an anti-CD20 antibody e.g., rituximab
• the human subject being treated for CRS is also administered a corticosteroid (e.g., hydrocortisone) and/or an anti-histamine (e.g., chlorphenamine) simultaneously, separately, or sequentially during the treatment with the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the subject is also administered an agent that inhibits IL-6 simultaneously, separately, or sequentially during the treatment with the anti-BDCA2 antibody or BDCA2-binding fragment thereof.
• the agent that inhibits IL-6 may be an anti-IL-6 antibody or IL6-binding fragment thereof, an IL6 receptor (IL6R) antagonist (e.g., tocilizumab or a soluble IL6R).
• IL6R IL6 receptor
• the anti- BDCA2 antibody or BDCA2-binding fragment thereof comprises the three heavy chain variable domain CDRs and the three light chain variable domain CDRs of BIIB059.
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises the amino acid sequences set forth in SEQ ID NOs.: 1-6.
• the anti- BDCA2 antibody or BDCA2-binding fragment comprises the amino acid sequences set forth in SEQ ID NOs.: 12-16.
• the anti-BDCA2 antibody or BDCA2- binding fragment comprises the amino acid sequences set forth in SEQ ID NOs.: 18-22.
• the anti-BDCA2 antibody or BDCA2-binding fragment comprises the amino acid sequences set forth in SEQ ID NOs.: 24-28.
• the anti-BDCA2 antibody or BDCA2-binding fragment thereof comprises a VH CDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 1 or 17, a VH CDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 2; and a VH CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.
• VL CDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.:4, a VL CDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO.: 5; and a VL CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO. 6.
• the anti- BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises (i) a VH domain that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of the VH domain of ⁇ 059 (SEQ ID NO:7), and/or (ii) a VL domain that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of the VL domain of B1IB059 (SEQ ID NO:8); or differs at least at 1 to 5 amino acid residues, but at fewer than 40, 30, 20, 15, or 10, residues, from SEQ ID NO:7 and/or SEQ ID NO:
• these anti-BDCA2 antibodies or BDCA2 -binding fragments (i) bind human or cynomolgus monkey BDCA2 but do not significantly bind BDCA2 from phylogenetic species below primates; and/or (ii) inhibit TLR7/TLR9-induced type I interferon and other cytokine or chemokine production by human pDCs; and/or (iii) mediate internalization of BDCA2 from the surface of pDCs; and/or (iv) downregulate CD32a and/or CD62L from the surface of pDCs; and/or (v) deplete pDCs in vitro by ADCC or CDC.
• the anti- BDCA2 antibody or antigen-binding fragment thereof selectively binds to the ectodomain of human BDCA2 and comprises (i) a HC that is at least 70%, 75%, 80%, 85%, 90%, 91%,
• the antibody formulation used in these studies comprised ⁇ 059, 10 mM citrate buffer, 140 mM Arg.HCl and 0.05% PS80.
• the formulation had a pH of 6.0.
• the highest concentration in these studies would be limited by viscosity and the limit imposed by the large volume subcutaneous pump: 50 cP.
• the viscosity was measured in the low concentration formulation ( Figure 1). It was found that the threshold viscosity was crossed between 225 and 250 mg/mL and 225 mg/mL was chosen as the highest concentration for anti-BDCA2 antibody formulations.
• Arg.HCl was again seen to be an advantage as it lowers the viscosity in a concentration-dependent manner.
• the Arg containing solutions did have a propensity for forming visible particles after incubation at 40°C.
• sucrose prevented the formation of visible particles (Table 3).
• Sucrose also lowered the counts of sub-visible-particulates ( Figure 5).
• Table 3 Viscosity (at time zero) and visible particles after incubation at 40°C for 1 month.
• Formulations were 20 mM Citrate pH 6.0, 0.05% PS80 with the additional excipients as shown.
• anti-BDCA2 antibody be formulated at 150 mg/mL.
• Formulation 1 150 mg/ml ⁇ 059, 20 mM citrate, 140 mM ArgJHCl, 0.05% PS80, pH 6.0; and Formulation 2: 150 mg/ml ⁇ 059, 20 mM histidine, 100 mM Arg.HCl, 3% sucrose, 0.05% PS80, pH 5.5. With this format it was possible to explore high concentration in two different formulations.
• the viscosity of Formulation 2 was then measured. As can be seen in Figure 8, the viscosity profile was amenable to incorporating this formulation into a device.
• the 10 cP threshold for an autoinjector was not crossed until -155 mg/mL, suggesting material of up to -140 mg/mL could go into this device.
• the 50 cP threshold had not been crossed at concentrations as high as 200 mg/mL, suggesting the possibility of going up to this concentration should a subcutaneous large volume injector be required.
• Dosing regimens were selected based on safety, pharmacokinetics (PK), PK-BDCA2 internalization relationship, and extrapolated inhibitory potency (concentration resulting in 90% inhibition of response [IC90]) of pDC IFNa production.
• BDCA2 target engagement as measured by BDCA2 internalization and reappearance, was observed in a dose-dependent manner across the dose range of 0.3 mg/kg to 20 mg/kg.
• EC90 values for BDCA2 internalization were derived from population-based PK and PD modeling with the mean value of 1.5 pg/mL.
• IC90 for IFNa inhibition was estimated from in vitro to in vivo extrapolation of BDCA2 internalization and IFNa inhibition.
• BIIB059 fixed doses of 50 mg, 150 mg and 450 mg subcutaneous (SC) administration every 4 weeks (Q4W) with an additional dose ("loading dose”) two weeks after
• this dose regimen with an additional dose of 450 mg at Week 2 and a bioavailability (F) of 0.45 is expected to result in cumulative exposure over 3 months comparable to that achieved by the single dose of 20 mg/kg IV for a 65-kg person, the highest dose tested in healthy volunteers.
• SC loading dose on Week 2 (Day 15 - i.e., 15 days after administration of the first dose) will be included.
• a high dose of 450 mg SC Q4W (for 12 weeks) and a loading dose at Week 2 is based on PK simulations using data with both SC and IV Q2W regimens, and the expectation that the 450 mg dose level will have adequate target (BDCA2) coverage to suppress pDC function, including the production of type I IFN, over the 12 weeks.
• BDCA2 target
• An anti-BDCA2 antibody drug product was formulated at a concentration of 150 mg/mL in 20 mM histidine, 100 mM Arg.HCl, 3% sucrose, 0.05% polysorbate-80 at pH 5.5.
• a formulation study was conducted to examine the stability of anti-BDCA2 antibody liquid formulations at concentrations above 150 mg/mL. Concentrations of 200, 225, and 240 mg/mL were examined in this study. Arginine and sucrose levels were also varied to understand the role of these excipients in the stability of high concentration formulations. Additionally, the pH of the formulations was increased to 6.0 or 6.5 to reduce the formation of basic species. A total of ten formulations was tested (see Table 6 for formulation compositions).
• Formulations 1 and 5 labeled 200/250/3/6 and 225/250/1/6, respectively, had the lowest aggregation at all the tested time points (0, 4, and 12 weeks) ( Figure 9).
• Figure 10 At 25°C ( Figure 10), 30°C ( Figure 11), and 40°C ( Figure 12), Formulation 1 consistently exhibited the lowest aggregate formation compared to other formulations.
• Figure 12 At 25°C ( Figure 10), 30°C ( Figure 11), and 40°C ( Figure 12), Formulation 1 consistently exhibited the lowest aggregate formation compared to other formulations. Thus, Formulation 1 was identified as the best performing formulation in this study. Linear modeling of the aggregation data from this study indicated that protein concentration, arginine concentration, and pH of the formulation significantly impacted aggregation.
• Anti-BDCA2 antibody ( ⁇ 059), SB4 (BENEPALI®), and anti-ct ⁇ f5 antibody (STX 200) were formulated according to the table below:
• Size exclusion HPLC (SEC) experiments were performed on a Waters Acquity UPLC instrument equipped with an Acquity UPLC BEH200 SEC analytical column coupled with a guard column. UV detection was performed at 280nm. A sample amount of 20 ⁇ g was injected on to the column by a constant flow rate of 0.35 mL/min mobile phase. Each sample ran for 10 min.
• SB4 and STX 200 were concentrated to 150 mg/ml in 10K centrifugal filters.
• the prepared solutions were plated in WebSeal plates with glass inserts, sealed and incubated at 25°C/60%RH and 40°C/75%RH for 3 months. Analysis of % HMW was performed by SEC at predetermined time points.
• Glutathione a tripeptide ( ⁇ -Glu-Cys-Gly) regulates disulfide bond formation.
• the reduced form (GSH) cleaves misbridged disulfide bonds and the oxidized form (GSSG) facilitates their formation.
• GSH + GSSG oxidized form
• Anti-BDCA2 antibody in presence of glutathione shows an initial reversible aggregation followed by an aggregation rate that is slower to the formulation with no glutathione at 2S°C ( Figure 17, left panel). Higher temperatures (40°C) add to the diversity of aggregation mechanisms, with conformational stability also coming into play ( Figure 17, right panel). Hence glutathione alone was not able to achieve similar reduction as at 25°C.
• Sucrose is a widely used excipient for protein stabilization. It is preferentially excluded from the protein surface, thus favoring its native conformation. Absence of sucrose in the anti-BDCA2 antibody formulation did not affect the aggregation profile ( Figure 18), further emphasizing the role of disulfide bond scrambling to control aggregation in BDCA2.
• STX 200 is an aglycosylated molecule, demonstrating poor conformational stability at higher temperatures. Hence, unfolding of the molecule exposes the thiol group making it more susceptible to crosslinking with the thiol in glutathione and promoting further aggregation. Glutathione also did not have any effect on the aggregation kinetics in SB4, a fusion protein at 2S°C, but facilitated faster aggregation at 40°C ( Figure 19).

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