Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
  • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
  • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• • 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
  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
  • A61K39/39591—Stabilisation, fragmentation
• • 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
  • A61K47/02—Inorganic compounds
• • 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
  • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
  • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
• • 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
  • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
• • 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
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/24—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
• • 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
  • A61K9/08—Solutions

Definitions

• the present disclosure relates to a pharmaceutical composition containing an anti-fractalkine antibody.
• Fractalkine (also referred to as "FKN”) is a membrane-bound chemokine expressed on the surface of vascular endothelial cells by inflammatory stimuli such as LPS, TNF- ⁇ , and IL-1.
• FKN receptor CX3CR1 binds to membrane-bound FKN without the intervention of selectins or integrins, causing strong cell adhesion.
• the secretory FKN shed from the membrane-bound FKN exhibits cell migration activity against NK cells, T cells, and monocytes having CX3CR1.
• clone 3A5-2 has high neutralizing activity, binding affinity and cross-reactivity between species with respect to hFKN, and the whole thereof is incorporated by reference to H3-2L4 (Patent Document 1, reference to Patent Document 1,) which is a humanized antibody of the antibody. ) Is given the international general name of Quetmolimab.
• the anti-FKN antibody containing H3-2L4 (also referred to as "Quetmolimab" in the present specification) needs to be administered at a high dose depending on the applicable disease.
• the concentration of the anti-FKN antibody in the preparation In order to increase the dose of the anti-FKN antibody without increasing the dose of the pharmaceutical composition to be administered, it is necessary to increase the concentration of the anti-FKN antibody in the preparation.
• increasing the concentration of the anti-FKN antibody increases the viscosity and cohesiveness, resulting in clogging of the syringe and increased sliding resistance (the force required at the time of administration increases, making self-administration difficult. ), The effect on the accuracy of quantification at the time of administration / manufacturing, and the decrease in filling speed at the time of manufacturing occur.
• a pharmaceutical composition containing an anti-fractalkine antibody comprises 200 mg / mL of the anti-fractalkine antibody and 100-400 mM basic amino acids.
• the anti-fractalkine antibody is: Heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS); A light chain variable region containing the amino acid sequence shown in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQFWSTPYTFGGGTKVEIK);
• the basic amino acid comprises at least one basic amino acid selected from arg
• composition according to any one of [1] to [5], which comprises one or more buffers selected from phosphoric acid, citric acid, succinic acid and salts thereof.
• Anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKG
• a high-concentration preparation of an anti-FKN antibody having a viscosity that is easy to handle is provided.
• anti-FKN antibody refers to the humanized anti-human fractalkine antibody H3-2L4, or an antibody functionally equivalent thereto.
• functionally equivalent antibody refers to antibody H3-2L4 and at least, or preferably all of, binding affinity for human FKN, neutralizing activity, cross-reactivity, and pharmacokinetics in blood. Refers to an antibody that is equivalent.
• antibody refers to a natural or artificial immunoglobulin molecule capable of recognizing and binding to a specific target or antigen via at least one antigen recognition site located in its variable region. ..
• the anti-FKN antibody according to the present disclosure is typically an antibody containing a constant region of a human IgG2 isotype.
• the anti-FKN antibody according to the present disclosure comprises a constant region of a human IgG2 isotype in which the Fc region comprises a mutation of V234A and / or G237A.
• the anti-FKN antibody is an antibody corresponding to a biosimilar when the antibody H3-2L4 is used as a reference product.
• Biosimilars refer to biopharmacy that has the same quality, safety, and efficacy as the preceding biopharmacy. For example, WHO guidelines (Guidelines on evaluation of similar Biotherapeutic Products (SBPs), Annex 2, Technical Report) It is defined as similar biotherapeutic products in Series No. 977, 2009). The guidelines define biosimilars to have the same primary structure as the reference product.
• the anti-FKN antibody exemplified above is appropriately modified (for example, modification of the antibody or partial of the amino acid sequence of the antibody) in order to maintain the function of the antibody or to add or improve the function of the antibody.
• Antibodies that have been substituted, added, or deleted) are also included in the antibodies according to the present disclosure. More specifically, in order to reduce the heterogeneity of the antibody produced by the antibody-producing cells, lysine (Lys) located at the carboxy-terminal (C-terminal) of the heavy chain is deleted by an artificial method such as gene modification. These antibodies are also included in the scope of the present disclosure.
• the anti-FKN antibody contained in the pharmaceutical composition according to the present disclosure does not necessarily have to have perfect uniformity, and as long as the pharmaceutical composition according to the present disclosure maintains the intended function, for example.
• a mixture of those in which lysine (Lys) located at the carboxy terminus (C-terminal) of the heavy chain is deleted and those in which lysine (Lys) is not deleted may be mixed.
• the anti-FKN antibody may be modified if desired.
• the anti-FKN antibody is (a) a three-dimensional structure of the amino acid sequence in the modified region, such as a sheet or helix conformation; (b) a charged or hydrophobic state of the molecule at the target site; or (c). ) It may be a modification that alters the effect of the modification on the maintenance of the volume of the side chain, or it may be a modification such that these changes are not clearly observed.
• Modification of the anti-FKN antibody may be achieved, for example, by substitution, deletion, addition, etc. of constituent amino acid residues.
• amino acid is used in its broadest sense and is a natural amino acid such as serine (Ser), aspartin (Asn), valine (Val), leucine (Leu), isoleucine (Ile), alanine (Ala). ), Tyr, glycine (Gly), lysine (Lys), arginine (Arg), histidine (His), aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), treonine (Thr), cysteine ( It includes not only Cys), methionine (Met), phenylalanine (Phe), tryptophan (Trp), proline (Pro), but also unnatural amino acids such as amino acid variants and derivatives.
• amino acids herein include, for example, L-amino acids; D-amino acids; chemically modified amino acids such as amino acid variants, amino acid derivatives; norleucine, ⁇ -alanine, and the like. It is naturally understood that amino acids such as ornithine, which are not constituents of proteins in vivo; and chemically synthesized compounds having the characteristics of amino acids known to those skilled in the art can be mentioned.
• unnatural amino acids include ⁇ -methyl amino acids ( ⁇ -methylalanine, etc.), D-amino acids (D-aspartic acid, D-glutamic acid, etc.), and histidine-like amino acids (2-amino-histidine, ⁇ -hydroxy-histidine, etc.).
• ⁇ -methyl amino acids ⁇ -methylalanine, etc.
• D-amino acids D-aspartic acid, D-glutamic acid, etc.
• histidine-like amino acids (2-amino-histidine, ⁇ -hydroxy-histidine, etc.).
• Homo histidine, ⁇ -fluoromethyl-histidine, ⁇ -methyl-histidine, etc. amino acids with excess methylene in the side chain (“homo” amino acids) and carboxylic acid functional amino acids in the side chain replaced by sulfonic acid groups
• examples thereof include amino acids (such as cysteine acid).
• Naturally occurring amino acid residues can be classified into the following groups, for example, based on general side chain properties: (1) Hydrophobicity: Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilicity: Cys, Ser, Thr; (3) Acid: Asp, Glu; (4) Basicity: Asn, Gln, His, Lys, Arg; (5) Residues affecting chain orientation: Gly, Pro; and (6) Aromatic: Trp, Tyr, Phe.
• Non-conservative substitution of the amino acid sequences constituting the anti-FKN antibody may be performed by exchanging an amino acid belonging to one of these groups with an amino acid belonging to the other group. More conservative substitutions may be made by exchanging amino acids belonging to one of these groups with other amino acids in the same group. Similarly, the amino acid sequence may be deleted or substituted as appropriate.
• the present disclosure relates to high concentration formulations of anti-FKN antibodies.
• the pharmaceutical composition according to the present disclosure is based on the finding that the anti-FKN antibody can be formulated at a high concentration by adding a specific basic amino acid, while having a low viscosity and excellent operability.
• the amount of the specific anti-FKN antibody may vary depending on the administration route and administration interval of the pharmaceutical composition according to the present disclosure, but the present disclosure particularly relates to a drug containing 200 mg / mL of the anti-FKN antibody. Regarding the composition.
• the basic amino acids referred to herein may include either L-form or D-form.
• the basic amino acid is an L-amino acid.
• Examples of the basic amino acids that can be used in the present disclosure include arginine, histidine, lysine, and ornithine.
• the basic amino acid may be used alone or in combination of two or more.
• basic amino acids comprising arginine, histidine, lysine, ornithine, or a combination thereof, are used.
• arginine, histidine, lysine, ornithine, or a combination thereof or more is used.
• arginine, histidine or lysine, or a combination thereof or more is used.
• arginine, histidine, or a combination of arginine and histidine is used as the basic amino acid.
• arginine is used as the basic amino acid.
• the concentration of a basic amino acid used varies depending on the type of basic amino acid selected, the route of administration, etc., and ranges from 25 mM to 500 mM, for example, 50 mM to 400 mM, 100 mM to 400 mM, 200 mM to 400 mM, Select from the range of 250 mM to 400 mM, 300 mM to 400 mM, 50 mM to 300 mM, 100 mM to 300 mM, 200 mM to 300 mM, 250 mM to 300 mM, 50 mM to 250 mM, 100 mM to 250 mM, 200 mM to 250 mM, 50 mM to 200 mM, or 100 mM to 200 mM. Can be done.
• the pharmaceutical product according to the present disclosure contains arginine at a concentration of about 50 mM or 50 mM, about 100 mM or 100 mM, about 200 mM or 200 mM, about 250 mM or 250 mM, about 300 mM or 300 mM, about 400 mM or 400 mM.
• the pharmaceutical product according to the present disclosure contains histidine at a concentration of about 50 mM or 50 mM, about 100 mM or 100 mM, about 200 mM or 200 mM, about 250 mM or 250 mM, about 300 mM or 300 mM, about 400 mM or 400 mM.
• the pharmaceutical compositions according to the present disclosure are about 50 mM or 50 mM in total, about 100 mM or 100 mM in total, about 200 mM or 200 mM in total, about 250 mM or 250 mM in total, about 300 mM or 300 mM in total, or It contains a combination of two or more amino acids selected from the group consisting of arginine, histidine, lysine and ornithine at a total concentration of about 400 mM or 400 mM. In another embodiment, the combination of the two or more amino acids is selected from the group consisting of arginine, histidine and lysine.
• the pharmaceutical compositions according to the present disclosure are about 50 mM or 50 mM in total, about 100 mM or 100 mM in total, about 200 mM or 200 mM in total, about 250 mM or 250 mM in total, about 300 mM or 300 mM in total, or It contains a combination of histidine and arginine at a total concentration of about 400 mM or 400 mM.
• the pharmaceutical composition according to the present disclosure is a combination of histidine at a concentration of about 50 mM or 50 mM and arginine at a concentration of about 250 mM or 250 mM, histidine at a concentration of about 100 mM or 100 mM and a concentration of about 200 mM or 200 mM.
• the buffer include phosphoric acid, citric acid, succinic acid, L-histidine, acetic acid, gluconic acid, tris, glycine or a salt thereof, or a combination thereof, and specific examples thereof include these acids.
• it is a base and / or a salt. If both an acid or a base and a salt are added, the salt can be a salt of the same or different acid or base.
• a pH adjusting agent commonly used by those skilled in the art such as potassium carbonate, sodium hydrogen carbonate, hydrochloric acid, and sodium hydroxide, may be used.
• the pharmaceutical composition in the present disclosure has a pH in the range of about 4.5 to about 6.5.
• the pH is in the range of pH 5.0 to 6.25, in the range of pH 5.25 to 6.00, or in the range of pH 5.5 to 5.8.
• the pharmaceutical composition according to the present disclosure has a pH of 5.8 or about 5.8.
• the concentration of the buffer can be from about 1 mM to about 600 mM depending on the type of buffer used and the desired isotonicity.
• the buffer is phosphoric acid and / or a salt thereof (eg, sodium phosphate) at a concentration of about 5 mM to about 40 mM, or about 10 mM to about 30 mM, or about 15 mM to about 25 mM. In another embodiment, the buffer is sodium phosphate at a concentration of about 25 mM or 25 mM.
• a salt thereof eg, sodium phosphate
• the buffer is citric acid and / or a salt thereof (eg, sodium citrate) at a concentration of about 5 mM to about 40 mM, or about 10 mM to about 30 mM, or about 15 mM to about 25 mM. In another embodiment, the buffer is citric acid at a concentration of about 25 mM or 25 mM.
• a salt thereof eg, sodium citrate
• the buffer is succinic acid and / or a salt thereof (eg, sodium succinate) at a concentration of about 5 mM to about 40 mM, or about 10 mM to about 30 mM, or about 15 mM to about 25 mM. In another embodiment, the buffer is succinic acid at a concentration of about 25 mM or 25 mM.
• a salt thereof eg, sodium succinate
• the buffer is L-histidine and / or a salt thereof at a concentration of about 5 mM to about 40 mM, or about 10 mM to about 30 mM, or about 15 mM to about 25 mM. In another embodiment, the buffer is L-histidine at a concentration of about 25 mM or 25 mM.
• the pharmaceutical composition according to the present disclosure can be formulated together with a stabilizer, a surfactant, a preservative and the like commonly used in the art.
• Stabilizers used in the present disclosure are, for example, carbohydrates or sugars or sugars, such as sucrose, which are approved by the authorities as suitable additives in pharmaceutical formulations.
• the concentration of stabilizer can be 15 to 250 mM, or 150 to 250 mM, or 200 mM.
• the pharmaceutical composition according to the present disclosure may contain a secondary stabilizer.
• Suitable examples of pharmaceutically acceptable surfactants used in the present disclosure are, but are not limited to, polyoxyethylene sorbitan fatty acid esters (Tween), polyethylene glycol glycols, polyoxyethylene stearate. , Polyoxyethylene alkyl ethers such as polyoxyethylene monolauryl ether, alkylphenylpolyoxyethylene ether (Triton-X), polyoxyethylene polyoxypropylene copolymer (Polyxamer, Pluronic), and sodium dodecyl sulfate (SDS). including.
• the most suitable polyoxyethylene sorbitan fatty acid esters are polysorbate 20 (Tween 20) and polysorbate 80 (Tween 80).
• the concentration of the surfactant is 0.01 to 0.1% (w / v), or 0.01 to 0.08% (w / v), or 0.025 to 0.075% (w / v). For example, 0.05% (w / v).
• polysorbate 80 is used as the surfactant.
• Preservatives used in the present disclosure include, but are not limited to, paraben, benzyl alcohol, sodium benzoate, phenol, benzalkonium chloride, thimerosal, chlorobutanol, benzoic acid, sodium bisulfite, sodium propionate and any of them. Including combinations or mixtures of.
• the pharmaceutical composition according to the present disclosure containing the above components can have a viscosity of less than 25 centipores (cP), preferably less than 20 cP.
• the viscosity (cP) refers to a value at 25 ° C.
• the viscosity (cP) of the pharmaceutical composition according to the present disclosure can be determined by a method known in the art, for example, a capillary viscometer method or a rotational viscometer method, using an apparatus or apparatus suitable for the method. .. In one embodiment, it is a capillary viscometer method, and the specific method used in the present specification, the apparatus used, and the like are described in Examples.
• composition according to the present disclosure containing the above components may be a pharmaceutical composition having low cohesiveness of the anti-FKN antibody.
• the present disclosure comprises adjusting a pharmaceutical composition comprising 200 mg / mL anti-FKN antibody to have a viscosity of less than 25 centimeter pores (cP) or less than 20 cP to prepare an anti-FKN antibody-containing pharmaceutical composition.
• a pharmaceutical composition comprising 200 mg / mL anti-FKN antibody to have a viscosity of less than 25 centimeter pores (cP) or less than 20 cP to prepare an anti-FKN antibody-containing pharmaceutical composition.
• the present disclosure also includes a method of reducing the viscosity to less than 25 centimeters (cP) or less than 20 cP in a pharmaceutical composition comprising a 200 mg / mL anti-FKN antibody.
• the pharmaceutical composition according to the present disclosure can be administered by a parenteral route such as subcutaneous, intravenous, or intramuscular.
• a parenteral route such as subcutaneous, intravenous, or intramuscular.
• the pharmaceutical composition according to the present disclosure is a pharmaceutical composition for subcutaneous administration.
• the pharmaceutical composition according to the present disclosure can be provided in a state of being pre-filled in a vial or an administration container (syringe).
• the pharmaceutical composition according to the present disclosure relates to FKN-related diseases such as rheumatoid arthritis, ulcerative colitis, Crohn's disease, autoimmune hepatitis, multiple sclerosis, and inflammatory diseases such as neurogenic pain. Can be used for treatment.
• the pharmaceutical composition according to the present disclosure comprises 200 mg / mL anti-FKN antibody, 50 mM arginine, 25 mM sodium phosphate, and 0.05 wt% polysorbate 80.
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDK
• the pharmaceutical composition according to the present disclosure comprises 200 mg / mL anti-FKN antibody, 100 mM arginine, 25 mM sodium phosphate, and 0.05 wt% polysorbate 80.
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNG
• the pharmaceutical composition according to the present disclosure comprises 200 mg / mL anti-FKN antibody, 300 mM arginine, 25 mM sodium phosphate, and 0.05 wt% polysorbate 80.
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNG
• the pharmaceutical composition according to the present disclosure comprises 200 mg / mL anti-FKN antibody, 400 mM arginine, 25 mM sodium phosphate, and 0.05 wt% polysorbate 80.
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNG
• the pharmaceutical composition according to the present disclosure comprises 200 mg / mL anti-FKN antibody, 250 mM lysine, 25 mM sodium phosphate, and 0.05 wt% polysorbate 80.
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNG
• the pharmaceutical composition according to the present disclosure comprises 200 mg / mL anti-FKN antibody, 250 mM histidine, 25 mM sodium phosphate, and 0.05 wt% polysorbate 80.
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKE
• the pharmaceutical composition according to the present disclosure comprises 200 mg / mL anti-FKN antibody, 250 mM arginine, 25 mM sodium phosphate, and 0.05 wt% polysorbate 80.
• the anti-fractalkine antibody is SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNG
• the term "comprise, include” is the presence of the described matter (member, step, element or number, etc.) unless the context clearly requires a different understanding. It is intended, and does not exclude the existence of other matters (members, steps, elements, numbers, etc.).
• the term “consist of” includes aspects described by the terms “consist of” and / or “consently of”.
• first and second are used to describe various elements, but these elements should not be limited by these terms themselves. These terms are used only to distinguish one element from the other, for example, the first element is referred to as the second element, and similarly, the second element is the first element. Is possible without departing from the scope of the present disclosure.
• Example 1 Preparation of humanized anti-human fractalkine antibody
• humanized anti-human fractalkine antibody H3-2L4 was used. Preparation of H3-2L4, including humanization, was performed as described in WO2011 / 052799. The H3-2L4 used after Example 2 is described in 1-1. And 1-2. Prepared by the method described in.
• Expression vector The expression vector for the humanized anti-human fractalkine antibody (H3-2L4) was prepared as follows. First, a signal sequence (SEQ ID NO: 3) was added to the N-terminal of the amino acid sequence (SEQ ID NO: 1) of the heavy chain variable region (H3-2) of the humanized anti-human fractalkine antibody (H3-2L4) at two locations. The amino acid sequence (SEQ ID NO: 4) of the constant region of human IgG2 into which the mutations (V234A and G237A) were inserted was added to the C-terminus (SEQ ID NO: 5).
• a signal sequence (SEQ ID NO: 6) was added to the N-terminal of the amino acid sequence (SEQ ID NO: 2) of the light chain variable region (L4) of the humanized anti-human fractalkine antibody (H3-2L4), and the signal sequence (SEQ ID NO: 6) was added to the C-terminal.
• the amino acid sequence (SEQ ID NO: 7) of the constant region of human Ig ⁇ was added (SEQ ID NO: 8).
• These amino acid sequences (SEQ ID NOs: 5 and 8) are converted into the optimum gene sequence for expression in CHO cells, and the recognition sequence and codon sequence of the restriction enzyme HindIII are stopped at the 5'end of the gene sequence at the 3'end.
• the sequences to which the recognition sequences of the codon and the restriction enzyme EcoRI were added were completely synthesized (SEQ ID NOs: 9 and 10).
• the sequences specified by SEQ ID NOs: 1 to 10 were as follows.
• H3-2 Amino acid sequence of heavy chain variable region (SEQ ID NO: 1) QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS
• L4 light chain variable region (SEQ ID NO: 2) DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK
• Amino acid sequence of constant region of human IgG2 with two mutations (V234A and G237A) inserted (SEQ ID NO: 4) ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQGNVFSCSVMHEALHNHYTQKSLSPGK
• the gene sequence encoding the fully synthesized heavy chain was excised with restriction enzymes HindIII and EcoRI, and inserted into the HindIII and EcoRI sites of the pEE6.4 vector (Lonza).
• the gene sequence encoding the fully synthesized light chain was excised with the restriction enzymes HindIII and EcoRI, and inserted into the HindIII and EcoRI sites of the pEE12.4 vector (Lonza).
• Each vector was cleaved with restriction enzymes NotI and PvuI, and vector fragments containing heavy and light chains were ligated to construct an expression vector.
• the antibody H3-2L4 obtained according to the above had a heavy chain and a light chain having the amino acid sequences shown in SEQ ID NOs: 11 and 12, respectively.
• amino acid sequences of the heavy chain variable region and the light chain variable region of the antibody H3-2L4 were as follows, respectively.
• H3-2L4 SEQ ID NO: 13
• H3-2L4 DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK
• amino acid sequences of the antibodies H3-2L4, CDR-H1 to CDR-H3, and CDR-L1 to CDR-L3 were as follows:
• Example 2 Examination of Formulation for Reducing Viscosity A 200 mg / mL preparation of the antibody H3-2L4 in the presence of 25 mM sodium phosphate, citric acid, succinic acid, or L-histidine, using various additives, 5 The viscosity was measured in the range of pH of .0 to 6.5 (Tables 1 to 4). An aqueous solution of HCl and NaOH was used to adjust the pH.
• Viscosity was measured using the capillary method. Specifically, the measurement was performed by a capillary method using a PA800plus (Beckman Coulter) equipped with a photodiode array detector. A base fused-silica capillary having an inner diameter of 50 ⁇ m was used. Capillaries were filled with viscosity standard solutions (5, 10 and 50 cP) or antibody H3-2L4 pharmaceutical sample at 80 psi for 5 minutes. A small amount of 0.02 wt% riboflavin was then loaded into the capillary and used as a dye to monitor the movement of the sample solution. A measuring instrument equipped with a sample vial at the capillary inlet was subjected to a constant pressure of 70 psi.
• Dye transfer to the detection window was detected at 445 nm.
• a standard curve was obtained by the migration time of riboflavin in viscosity standard solutions (5, 10 and 50 cP). The viscosity was determined from the migration time when the peak of riboflavin moved in the capillary filled with the sample. All measurements were performed at 25 ° C. Results
• Addition of basic amino acids (L-arginine, L-histidine, lysine, and ornithine) significantly reduced the viscosity (Table 1, Nos. 2-9).
• the preparations using L-histidine and L-arginine had a large effect of reducing the viscosity (Table 1).

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