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WO2025037336A1 - Formulations pharmaceutiques d'anticorps anti-cd 20 et leurs procédés de préparation - Google Patents

Formulations pharmaceutiques d'anticorps anti-cd 20 et leurs procédés de préparation Download PDF

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Publication number
WO2025037336A1
WO2025037336A1 PCT/IN2024/051485 IN2024051485W WO2025037336A1 WO 2025037336 A1 WO2025037336 A1 WO 2025037336A1 IN 2024051485 W IN2024051485 W IN 2024051485W WO 2025037336 A1 WO2025037336 A1 WO 2025037336A1
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Prior art keywords
antibody
buffer
formulation
composition
surfactant
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Inventor
Murali JAYARAMAN
Anshu DEOGAM
Siddharth SONI
Ravi Kumar MARIKANTY
Sucheta RAJARAMAN
Indra Kumar SIGIREDDI
Suman LABALA
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Dr Reddys Laboratories Ltd
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Dr Reddys Laboratories Ltd
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Publication of WO2025037336A1 publication Critical patent/WO2025037336A1/fr
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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • 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
    • C07K16/2887—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD20
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered

Definitions

  • the present invention relates to the field of therapeutic antibody formulations. More specifically, the present invention relates to the field of stable formulations of antibodies and antigen-binding fragments against human CD-20 antigen and method for preparing the same.
  • mAbs Recombinant therapeutic monoclonal antibodies require a mammalian expression system (for example CHO cells) to be produced.
  • the molecular weight of a monomer of a therapeutic mAb ranges between 140-160 kDa, which is usually higher than other chemically synthesized oligopeptides.
  • mAb therapeutics are either lyophilized or supplied as liquid -based formulations for parenteral transmission to achieve highest level of bioavailability in patients.
  • the protein instability and degradation as a result of the harsh pH and salt conditions in the downstream process is one of the challenges for the maximum recovery of the purified product.
  • other factors such as protein’s intrinsic instability, aggregate formation due to self-association, insolubility and viscosity are the other major constraints for formulating the therapeutic mAbs or mAb based therapeutics. Chemical factors like oxidation, hydrolysis, disulfide exchange, deamidation accelerate the degradation of therapeutic mAbs.
  • the optimization of the mAb formulation is performed in such a way that the highest concentration is achieved at the minimum achievable volume for parenteral route of administration without affecting the overall quality of the mAb.
  • the selection of excipients in the formulation buffer depends on the compatibility with the protein, concentration of the protein, buffer, other excipients present and also the concentration of the excipients. Therefore, the correct screening and selection of the formulation buffers is a challenge to obtain the accurate formulation of the mAb in the buffer which is free from aggregate formation, free of colloidal particles, lesser acidic/basic variants.
  • the present invention discloses a pharmaceutical formulation of an anti-CD-20 antibody formulation, wherein the formulation comprises anti-CD20 antibody, buffer, stabilizer and surfactant and pH of the formulation is about 5.0 to about 6.5.
  • the invention comprises obtaining a stable anti-CD20 antibody formulation in succinate containing buffer, in particular, sodium succinate buffer.
  • the invention discloses a method of preparing/obtaining a stable anti- CD20 antibody composition wherein the method comprises, preparing sodium succinate buffer in a specific way and obtaining the anti-CD20 antibody composition in the prepared sodium succinate buffer.
  • the invention particularly discloses a method of reducing light induced aggregation or fragmentation of anti-CD20 antibody in it’s composition, wherein the method comprises obtaining anti-CD20 antibody in a buffer composition comprising sodium succinate buffer or histidine buffer or it’s derivatives.
  • the invention particularly discloses a method of reducing basic variants formation of anti-CD20 antibody in the anti-CD20 antibody composition, wherein the method comprises obtaining ant-CD20 antibody in a buffer composition comprising sodium succinate buffer or histidine buffer or it’s derivatives.
  • the disclosed method specifically controls pyroglutamate formation and/or lysine variants formation and/or methionine oxidation or succinimide formation of an anti-CD20 antibody thereby reduces formation of basic variants in anti-CD20 antibody composition.
  • the invention particularly discloses a method of reducing fragmentation of anti-CD20 antibody in the anti-CD20 antibody composition, wherein the method comprises obtaining ant-CD20 antibody in a buffer composition comprising sodium succinate buffer or histidine buffer or it’s derivatives. Additionally, the disclosed method controls aggregation of anti-CD20 antibody in it’s composition.
  • the disclosed formulations and the methods of the present invention stabilizes anti- CD20 antibody from lower to higher concentration (i.e; from 10 mg/ml to 200 mg/ml) and suitable for intravenous as well as subcutaneous formulations.
  • the pH of the formulation of the present invention can range from about 5.0 to about 6.5.
  • the disclosed formulations and the methods of the invention exhibit stability under at least one of the following accelerated conditions that includes a temperature ranging from 25 °C to 40 °C and for a period of time ranging from 1 day to 28 days/4 weeks.
  • the antibody in the said formulation is stable and maintains 98% or more (> 98 %) of monomeric content of the antibody in the formulation even after storage for two to four weeks at 40 °C.
  • the present invention discloses a pharmaceutical formulation of an anti-CD20 antibody.
  • the invention discloses a pharmaceutical formulation of an anti CD- 20 antibody comprising:
  • the invention discloses a pharmaceutical formulation of an anti-CD20 antibody comprising:
  • the disclosed formulation further comprises one or more pharmaceutically acceptable excipients or stabilizers.
  • the formulation comprises one or more pharmaceutically acceptable excipients which includes sugars, amino acids, or polymers or salts.
  • the said buffer is a succinate buffer or an acetate buffer or a citrate buffer or a histidine buffer or a phosphate buffer or combination thereof.
  • the succinate buffer is a sodium succinate buffer.
  • the buffer is sodium succinate buffer, that is prepared in a specific way with the following steps; a) dissolution of succinic acid in water to obtain succinic acid solution b) preparation of sodium hydroxide solution by dissolving sodium hydroxide in water c) mixing of sodium hydroxide solution to succinic acid solution in a particular molar ratio to obtain sodium succinate buffer.
  • the molar ratio of succinic acid to sodium hydroxide is 1 : 1 to 1:3.
  • the invention discloses a pharmaceutical formulation of an anti-CD20 antibody comprising:
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising: i) 10 mg/ml to 200 mg/ml an anti-CD20 antibody, ii) 10-50 mM sodium succinate buffer, iii) one or more stabilizers comprise mannitol, or trehalose, or sucrose or sorbitol; or sodium chloride or derivatives thereof and; iv) a surfactant.
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising: i) 120 mg/ml an anti-CD20 antibody, ii) 10-50 mM sodium succinate buffer, iii) one or more stabilizers comprise mannitol, or trehalose, or sucrose or sorbitol; or sodium chloride or derivatives thereof and; iv) a surfactant.
  • the succinate buffer or it’s derivatives or salts or combination thereof is a sodium succinate buffer.
  • the said buffer is prepared in a specific way in a specific molar ratio of succinate and sodium hydroxide.
  • the specific molar ratio is in between 1:1 to 1:3.
  • the buffer composition may further contain at least one or more pharmaceutically acceptable excipients/stabilizer. The said excipients can be added during preformulation and/or formulation stage of the antibody production.
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising:
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising: i) 10 mg/ml to 200 mg/ml an anti-CD20 antibody, ii) 10-50 mM histidine buffer, iii) one or more stabilizers comprise mannitol, or trehalose, or sucrose or sorbitol; or sodium chloride or derivatives thereof and; iv) a surfactant.
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising: i) 120 mg/ml an anti-CD20 antibody, ii) 10-50 mM histidine buffer, iii) one or more stabilizers comprise mannitol, or trehalose, or sucrose or sorbitol; or sodium chloride or derivatives thereof and; iv) a surfactant.
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising: i) 10 mg/ml to 200 mg/ml an anti-CD20 antibody, ii) 10-50 mM histidine acetate buffer, iii) one or more stabilizers comprise mannitol, or trehalose, or sucrose or sorbitol; or sodium chloride or derivatives thereof and; iv) a surfactant.
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising: v) 10 mg/ml to 200 mg/ml an anti-CD20 antibody, vi) 10-50 mM histidine-succinate buffer, vii)one or more stabilizers comprise mannitol, or trehalose, or sucrose or sorbitol; or sodium chloride or derivatives thereof and; viii) a surfactant.
  • the histidine buffer or it’s derivative or salts or combinations thereof is a histidine buffer or a histidine-citrate buffer or a histidinephosphate buffer.
  • the histidine buffer composition may also further contain at least one pharmaceutically acceptable excipients/stabilizer. The said composition can be added during pre-formulation and formulation stage of the antibody production.
  • the invention discloses a pharmaceutical formulation of an anti- CD20 antibody comprising: i) 10 mg/ml to 200 mg/ml an anti-CD20 antibody, ii) 10-50 mM citrate-phosphate buffer, iii) one or more stabilizers comprise mannitol, or trehalose, or sucrose or sorbitol; or sodium chloride or derivatives thereof and; iv) a surfactant.
  • the invention discloses formulations of Table-1 as disclosed below.
  • Table 1 Anti-CD20 antibody formulation composition
  • the invention discloses a method of obtaining a stable formulation of anti-CD20 antibody, wherein the method comprises, a. dissolution of succinic acid in water to obtain succinic acid solution b. preparation of sodium hydroxide solution by dissolving sodium hydroxide in water c. mixing of sodium hydroxide solution to succinic acid solution in a specific molar ratio to obtain sodium succinate buffer and; d. preparation of anti-CD-20 antibody in a buffer composition comprising the said sodium succinate buffer; wherein the antibody solution obtained from step d) is stable.
  • the molar ratio between succinic acid to sodium hydroxide is 1: 1 to 1:3 to obtain sodium succinate buffer composition.
  • the molar ratio between succinic acid to sodium hydroxide is 1:1, 1: 1.1; 1: 1.2; 1:1.3, 1: 1.4; 1: 1.5; 1:1.6; 1: 1.7; 1: 1.8; 1: 1.9; 1:2; 1:2.1; 1:2.1; 1:2.2; 1:2.3; 1:2.4; 1:2.5; 1:2.6; 1:2.7; 1:2.8; 1:2.9 and 1:3.
  • obtain sodium succinate buffer composition obtained by reacting of sodium hydroxide.
  • the concentration of antibody is about 10 mg/ml to 200 mg/ml.
  • the buffer composition may additionally comprise one or more pharmaceutically acceptable excipients.
  • the pharmaceutically acceptable excipients include sugar, polyol, polymers, amino acid or it’s derivatives, salts.
  • the invention discloses a method of preparing a buffer composition to obtain a stable formulation of anti-CD20 antibody, wherein the method comprises, a) dissolution of L-histidine and L-histidine hydrochloride in water in a specific molar ratio to obtain histidine buffer b) preparation of anti-CD20 antibody in a buffer composition comprising the said histidine buffer.
  • the molar ratio between L-histidine to L-histidine hydrochloride is between 1:3 to 1:5.
  • the invention discloses a method to prepare a pharmaceutical formulation of anti-CD20 antibody comprising formulating an anti-CD20 antibody in a buffer composition comprising sodium succinate buffer, sugar and/or surfactant.
  • the invention discloses a method of controlling aggregation of an anti-CD-20 antibody, in an anti-CD20 antibody composition, wherein the method involves preparation of anti-CD20 antibody in succinate buffer or histidine buffer or it’s derivatives or salts or combinations thereof.
  • the invention discloses a method of controlling agitation induced aggregation of an anti-CD-20 antibody, in an anti-CD20 antibody composition, wherein the method involves preparation of anti-CD20 antibody in succinate buffer or histidine buffer or it’s derivatives or salts or combinations thereof.
  • the invention discloses a method of controlling light induced aggregation of an anti-CD-20 antibody, in an anti-CD20 antibody composition, wherein the method involves preparation of anti-CD20 antibody in succinate buffer or histidine buffer or it’s derivatives or salts or combinations thereof.
  • the buffer composition may not require methionine or anti-oxidant or chelating agent to control light induced aggregation of anti-CD20 antibody.
  • the invention discloses a method to obtain a stable anti-CD20 formulation against light induced aggregation of anti-CD20 antibody comprising; formulating/preparation of anti-CD20 antibody in a buffer composition comprising sodium succinate buffer, sugar and surfactant.
  • the invention discloses a method to obtain a stable anti-CD20 formulation against light induced aggregation of anti-CD20 antibody comprising; formulating/preparation of anti-CD20 antibody in a buffer composition comprising histidine buffer, sugar and surfactant.
  • the invention discloses a method to obtain a stable anti-CD20 antibody formulation against light induced aggregation/degradation of anti-CD20 antibody comprising; formulating/preparation of anti-CD20 antibody in a buffer composition comprising sodium succinate buffer and one or more stabilizers.
  • sodium succinate is prepared in a specific way in a specific molar ratio of succinate and sodium hydroxide.
  • the specific molar ratio is in between 1: 1 to 1:3.
  • the formulation additionally /optionally comprises surfactant.
  • the invention discloses a method to obtain a stable anti-CD20 antibody formulation against light induced aggregation/degradation of anti-CD20 antibody comprising; formulating/preparation of anti-CD20 antibody in a buffer composition comprising histidine buffer and one or more stabilizers.
  • the formulation additionally /optionally comprises surfactant.
  • the invention discloses a method of controlling fragmentation of an anti-CD-20 antibody, in an anti-CD20 antibody composition, wherein the method involves formulating/preparation of anti-CD20 antibody in a buffer composition comprising sodium succinate buffer.
  • the buffer composition further comprises sugar and surfactant.
  • the invention discloses a method to obtain a stable formulation against light induced fragmentation of anti-CD20 antibody comprising; formulating/preparation of anti-CD20 antibody in a buffer composition comprising sodium succinate buffer.
  • the buffer composition further comprises sugar and surfactant.
  • sodium succinate is prepared in a specific way in a specific molar ratio of succinate and sodium hydroxide.
  • the specific molar ratio is in between 1: 1 to 1:3.
  • the invention discloses a method to obtain a stable formulation against light induced fragmentation of anti-CD20 antibody comprising; formulating anti-CD20 antibody in a buffer composition comprising histidine buffer, sugar and surfactant.
  • the invention discloses a method of controlling charge variants formation of an anti-CD-20 antibody, in an anti-CD20 antibody composition, wherein the method comprises preparation of anti-CD20 antibody in a buffer composition comprising succinate buffer or histidine buffer or it’s derivatives or salts or combinations.
  • the invention discloses a method of controlling deamidation of an anti-CD-20 antibody, in an anti-CD20 antibody composition, wherein the method comprises preparation of anti-CD20 antibody in a buffer composition comprising succinate buffer or histidine buffer or it’s derivatives or salts or combinations.
  • the invention discloses a method of controlling formation of basic variants in an anti-CD20 antibody composition wherein the method comprises preparation of anti-CD20 antibody in a buffer composition comprising succinate buffer or histidine buffer or it’s derivatives or salts or combinations. The method specifically controls formation of pyroglutamate variants, succinimide variants and lysine variants thereby reduces formation of basic variants.
  • the invention discloses a method of controlling formation of pyroglutamate variants in an anti-CD20 antibody composition wherein the method comprises preparation of anti-CD20 antibody in a buffer composition comprising succinate buffer or histidine buffer or it’s derivatives or salts or combinations.
  • the invention discloses a method of controlling formation of lysine variants in an anti-CD20 antibody composition, wherein the method comprises preparation of anti-CD20 antibody in a buffer composition comprising succinate buffer or histidine buffer or it’s derivatives or salts or combinations.
  • the invention discloses a method of controlling formation of succinimide variants in an anti-CD20 antibody composition, wherein the method comprises preparation of anti-CD20 antibody in a buffer composition comprising succinate buffer or histidine buffer or it’s derivatives or salts or combinations.
  • the invention discloses a method of imparting colloidal stability to an anti-CD-20 antibody, in an anti-CD20 antibody composition, wherein the method comprises preparation of anti-CD20 antibody in a buffer composition comprising succinate buffer or histidine buffer or it’s derivatives or salts or combinations.
  • the invention discloses, a method of controlling oxidation of Met34 and Mets3 of heavy chain of anti-CD20 antibody in a pharmaceutical composition of anti-CD20 antibody, wherein the method comprises preparation of the antibody composition in a buffer composition comprising succinate buffer or histidine buffer, sugar, and surfactant.
  • the invention discloses a method of controlling visible particle formation in an anti-CD20 antibody composition, wherein the method comprises preparation of the antibody composition in a buffer composition comprising succinate buffer or histidine buffer, sugar, and surfactant.
  • the invention discloses a method of controlling sub-visible particle formation in an anti-CD20 antibody composition wherein the method comprises preparation of the antibody composition in a buffer composition comprising succinate buffer or histidine buffer, sugar, and surfactant.
  • sodium succinate is prepared in a specific way in a specific molar ratio of succinate and sodium hydroxide.
  • the specific molar ratio is in between 1: 1 to 1:3.
  • the pH of the disclosed formulation of the present invention is in the range from about 5.0 to about 6.5.
  • the pH of the disclosed formulation of the present invention is 5.5 ⁇ 0.3.
  • the disclosed formulations of the present invention stabilize an anti-CD20 antibody at various concentration levels i.e., from about 10 mg/ml to about 200 mg/ml and are suitable for intravenous as well as subcutaneous routes of administration. Further, viscosity of the anti- CD20 antibody formulations is less than 20 cP, specifically, less than 10 cP.
  • the one or more pharmaceutically acceptable excipients/stabilizers is a sugar, polyol, amino acid, or salt.
  • polyol is mannitol or sorbitol.
  • the salt is sodium chloride
  • the surfactant is polysorbate or poloxamer.
  • the sodium succinate buffer is prepared by following steps: a. dissolution of succinic acid in water b. preparation of sodium hydroxide solution by dissolving sodium hydroxide in water c. mixing of sodium hydroxide to succinic acid solution in a particular molar ratio to obtain sodium succinate buffer.
  • the molar ratio between succinic acid to sodium hydroxide is 1: 1 to 1:3.
  • the claimed anti-CD20 antibody formulations of the invention exhibit stability under at least one of the following conditions, wherein the temperature range from 25 °C to 50 °C for a period of time which includes from
  • the formulation may not require an anti-oxidant or chelating agent to stabilize anti-CD20 antibody.
  • the chelator includes ethylenediamine tetraacetic acid (EDTA) or ethylene glycol-bis(P-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) or diethylenetriamine pentaacetate (DTPA) or like.
  • EDTA ethylenediamine tetraacetic acid
  • EGTA ethylene glycol-bis(P-aminoethyl ether)-N,N,N',N'-tetraacetic acid
  • DTPA diethylenetriamine pentaacetate
  • the anti-CD20 antibody is a chimeric or humanized or human anti-CD20 antibody or conjugates or fusion proteins thereof.
  • the anti-CD20 antibody is either a type-I or type-
  • the anti-CD20 antibody is rituximab, ofatumumab, obinutuzumab, ocelizumab or ubilituximab or conjugates thereof.
  • conjugates is a drug or an antibody or any other therapeutic molecule.
  • the fusion proteins include an anti-CD 20 antibody or antigen fragment thereof fused to another protein.
  • the concentration of the antibody in the formulation is about 10 mg /ml to about 200 mg/ml.
  • the concentration of the antibody in the formulation is 10 mg/ml, or 25 mg/ml, or 50 mg/ml, or 60 mg/ml, or 70 mg/ml, or 80 mg/ml, 90 mg/ml, or 100 mg/ml, or 110 mg/ml, or 120 mg/ml, or 150 mg/ml or 160 mg/ml, or 170 mg/ml or 175 mg/ml or 180 mg/ml or 190 mg/ml or 195 mg/ml or 200 mg/ml.
  • the anti-CD20 antibody formulation is stable and contains less than 1.5 % of high molecular weight (HMW) species in the formulation, even after storage at 40 °C for four weeks.
  • HMW high molecular weight
  • the anti-CD20 antibody formulation is stable and contains less than 2% of fragments in the formulation, even after storage at 40 °C for four weeks.
  • the anti-CD20 antibody maintains at least 90% of monomeric content of the antibody after storage at 40 °C for four weeks
  • the anti-CD20 antibody formulation’s osmolality is less than 600 mOsm/kg, preferably less than 300 mOsm/kg.
  • the anti-CD20 antibody formulations disclosed in the invention are biologically active.
  • the formulation of anti-CD20 antibody is a stable liquid (aqueous) formulation, which can be used for parenteral administration.
  • Parenteral administration includes intravenous, subcutaneous, intra peritoneal, intramuscular administration or any other route of delivery generally considered to be falling under the scope of parenteral administration and as is well known to a skilled person.
  • the stable liquid/aqueous formulation is suitable and can be lyophilized as lyophilized powders. Further, the lyophilized formulation of anti-CD20 antibody can be reconstituted with appropriate diluent to achieve the liquid formulation suitable for administration.
  • liquid/aqueous anti-CD20 antibody are compatible with lyophilization process and the lyophilization process does not impact quality attributes of the antibody.
  • a vial, pre-filled syringe or autoinjector device or any other suitable device comprising any of the subject formulations described herein.
  • the aqueous formulation, stored in the vial or pre-filled syringe or an auto injector device comprise anti-CD20 antibody, succinate buffer or acetate buffer or citrate buffer or histidine buffer or it’s derivatives or combination thereof, sugar and surfactant.
  • antibody refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, or an antigen -binding portion thereof.
  • the “antibody” as used herein encompasses whole antibodies or any antigen binding fragment (i.e., “antigen-binding portion”) or fusion protein thereof.
  • stable formulation refers to the formulation wherein the antibody therein retains its physical stability and/or chemical stability and/or biological activity upon storage.
  • An antibody "retains its physical stability” in a pharmaceutical formulation if it shows substantially no signs of aggregation, precipitation and/or denaturation upon visual examination of color and/or clarity, or as measured by UV light scattering or by size exclusion chromatography.
  • An antibody is said to “retain its chemical stability” in a pharmaceutical formulation when its shows no or minimal formation of product variants which may include variants as a result of chemical modification of antibody of interest such as deamination, oxidation etc.
  • Analytical methods such as ion exchange chromatography and hydrophobic ion chromatography may be used to investigate the chemical product variants.
  • the term ‘monomer’ as used herein describes antibodies consisting of two light chains and two heavy chains.
  • the monomer content of an antibody composition is typically analyzed by size exclusion chromatography (SEC).
  • SEC size exclusion chromatography
  • HMW high molecular weight
  • aggregates that may include dimers, multimers, etc., elute first, followed by monomer, and the clipped antibody variants or degradants may be eluted last.
  • the aggregate peak or the degradant peaks may not elute as a baseline separated peaks but instead as a shoulder or abnormal broad peaks.
  • main peak refers to the peak that elutes in abundance (major peak) during a cation exchange chromatography.
  • the peak that elutes earlier than the main peak, during a cation exchange chromatography, with a charge that is acidic relative to the main peak is termed acidic variant peak.
  • the peak that elutes later than the main peak, during a cation exchange chromatography, with a charge that is relatively basic than the main peak is termed as basic variant peak.
  • the main peak content can be determined by Ion exchange chromatography (IEC). There are two modes of IEC available viz., cation and anion exchange chromatography.
  • Positively charged molecules bind to anion exchange resins while negatively charged molecules bind to cation exchange resins.
  • acidic variants elute first followed by the main peak and thereafter lastly the basic variants will be eluted.
  • the acidic variants are a result of antibody modifications such as deamidation of asparagine residues.
  • the basic variants are a result of incomplete removal of C-terminal lysine residue(s). In general, in an antibody a lysine residue is present at the C-terminal end of both heavy and light chain.
  • K2 variant An antibody molecule containing lysine at both heavy and light chain is referred to as K2 variant
  • the antibody molecule containing lysine residue at either one of heavy and light chain is referred to as KI variant and antibody molecule having none is KO molecule.
  • Carboxypeptidase B (CP-B enzyme) enzyme acts on the C-terminal lysine residues present on K2 and KI variants and thus converting them as KO molecules.
  • the IEC analysis can be carried out for samples digested with carboxypeptidase B (CP-B) enzyme.
  • CP-B carboxypeptidase B
  • compositions refer to the additives or carriers, which may contribute to stability of the antibody in formulation.
  • the excipients may encompass stabilizers and tonicity modifiers.
  • stabilizers and tonicity modifiers include, but not limited to, salts, surfactants, and derivatives and combination thereof.
  • the at least one stabilizer in a pharmaceutical formulation of the present invention can be a polyethylene glycol (PEG), P- cyclodextrin or equivalents thereof.
  • sugar/s as used herein includes organic compounds having general formula of all carbohydrates of the general formula Cn(H20)n.
  • Sugars can be referred to monosaccharides, disaccharides, and polysaccharides.
  • examples of sugars include, but are not limited to, sucrose, trehalose, glucose, dextrose, raffinose and others.
  • polyol or “sugar alcohol” as used herein includes an organic compound containing multiple hydroxyl groups. Examples of polyols include mannitol, sorbitol, xylitol etc.,
  • Surfactant refers to pharmaceutically acceptable excipients used to protect the protein formulations against various stress conditions, like agitation, shearing, exposure to high temperature etc.
  • suitable surfactants include but are not limited to polyoxyethylensorbitan fatty acid esters such as Tween 20TM or Tween 80TM, polyoxyethylene-polyoxypropylene copolymer (e.g. Poloxamer, Pluronic), sodium dodecyl sulphate (SDS) and the like or combination thereof.
  • salts include, but not limited to, sodium chloride, potassium chloride, magnesium chloride, sodium thiocyanate, ammonium thiocyanate, ammonium sulfate, ammonium chloride, calcium chloride, zinc chloride and/or sodium acetate.
  • opalescence or “opalescent appearance” refers to the degree of turbidity detected in a solution, e.g., a protein preparation, as a function of the concentration of one or more of the components in the solution, e.g., protein and/or salt concentration.
  • the degree of turbidity can be calculated by reference to a standard curve generated using suspensions of known turbidity. Reference standards for determining the degree of turbidity for pharmaceutical compositions can be based on the United States Pharmacopeia or European Pharmacopeia criteria.
  • first Formazine solution has been prepared by mixing equal volumes of a hydrazine sulfate solution and hexamethylenetetramine solution and then diluted to prepare various reference opalescence standards.
  • the opalescence standards includes ROS-I, ROS-II, ROS-III and ROS-IV.
  • Nephelometry is a turbidometric method used to detect the presence of soluble aggregates or to indicate opalescence. The output is listed in terms of nephelometric turbidity units (NTUs).
  • Pre-formulation steps refers to any or multiple steps performed before formulating the protein into a therapeutic product. Examples of such steps include, chromatography, filtration, (ultrafiltration, sterile filtration, nano filtration, diafiltration, tangential flow filtration, depth filtration), or any other steps performed to concentrate the protein or to exchange the buffer to a different/suitable buffer.
  • the filtration steps mentioned herein may be performed in a tangential flow filtration mode.
  • Formulation steps refers to steps which are followed after the downstream chromatographic and filtration steps to prepare drug product from drug substance, the latter obtained from the pre-formulation steps.
  • chelators/chelating agents refers to a compound which can form at least one bond with a metal atom.
  • a chelating agent is typically a multidentate ligand that can be used in compositions as a stabilizer to complex with species, which might otherwise promote instability.
  • Exemplary chelating agents include aminopolycarboxylic acids, hydroxyaminocarboxylic acids, N- substituted glycines, 2- (2-am ino-2-oxocthyl) aminoethane sulfonic acid (BES), deferoxamine (DEF), niacinamide, desoxycholates, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), nitrilotriacetic acid (NTA), N-2-acetamido-2- iminodiacetic acid (ADA), bis(am inoethyl)glycolether, N,N,N',N'-tetraacetic acid (EGTA), trans- diaminocyclohexane tetraacetic acid (DCTA), N- hydroxyethyliminodiacetic acid (HIMDA), N,N-bis- hydroxy ethylglycine (bicine), N- (trishy
  • antioxidant refers to an agent that inhibits the oxidation of other molecules and is not part of buffer component.
  • antioxidants herein include citrate, methionine, lipoic acid, uric acid, glutathione, tocopherol, carotene, lycopene, cysteine, phosphonate compounds, e.g., etidronic acid, desferoxamine and malate.
  • an anti-CD20 antibody, ocrelizumab, suitable for storage in the present pharmaceutical composition is produced by standard methods known in the art.
  • ocrelizumab is prepared by recombinant expression of immunoglobulin light and heavy chain genes in a mammalian host cell such as Chinese Hamster Ovary cells.
  • the expressed ocrelizumab is harvested and the crude harvest is subjected to standard downstream process steps that include purification, filtration and optionally dilution or concentration steps.
  • the crude harvest of ocrelizumab may be purified using standard chromatography techniques such as affinity chromatography, ion-exchange chromatography and combinations thereof.
  • the purified ocrelizumab solution can additionally be subjected to one or more filtration steps, and the solution obtained is subjected to further formulation studies.
  • Example 1 Anti-CD20 antibody formulations.
  • Purified ocrelizumab antibody sample approximately 8 mg/ml in acetate buffer back ground was obtained from downstream chromatographic step.
  • the antibody containing sample was buffer exchanged with different buffer compositions comprising succinate buffer (i.e., succinic acid and di sodium succinate hexahydrate), histidine buffer (L histidine and L- histidine hydrochloride), histidine- succinate buffer (L histidine and disodium succinate hexahydrate) and citrate phosphate.
  • succinate buffer i.e., succinic acid and di sodium succinate hexahydrate
  • histidine buffer L histidine and L- histidine hydrochloride
  • histidine- succinate buffer L histidine and disodium succinate hexahydrate
  • citrate phosphate citrate phosphate
  • the samples were concentrated to a concentration ranging between 40 mg/ml to 120 mg/ml and the concentration was further adjusted to 30 mg/ml with formulation buffer having pH 5.3 ⁇ 0.3 comprising excipients such as sugar at a concentration range of 10 mg/ml to 60 mg/ml and surfactant at a concentration range of 0.1 to 0.5 mg/ml. Details of the formulation are given in Table 2.
  • LMW low molecular weight
  • HMW high molecular weight species
  • SEC size exclusion chromatography
  • Table 2 Compositions of anti-CD20 antibody formulations prepared as per Example- 1
  • Table 5 IEX data on anti-CD20 antibody formulations prepared as per examp
  • Table 6 Quality attributes of anti-CD 20 antibody formulations prepared as per Example- 1.
  • Example 2 High concentration Anti-CD20 antibody formulations.
  • Purified rituximab antibody sample approximately 8 mg/ml in acetate buffer background was obtained from downstream chromatographic step.
  • the antibody containing sample was buffer exchanged with Histidine acetate.
  • Post which, the samples were concentrated to a concentration ranging between 40 mg/ml to 190 mg/ml with formulation buffer having pH 6.5 ⁇ 0.3 comprising excipients such as sugar at a concentration range of 80 mg/ml to 150 mg/ml, surfactant at a concentration range of 0.1 to 0.7 mg/ml, amino acid at a range of 5 mg/ml to 12 mg/ml, polyethylene glycol (PEG) and diethylenetriamine pentaacetate (DTP A). Details of the formulation are given in Table 8.
  • LMW low molecular weight
  • HMW high molecular weight species
  • SEC size exclusion chromatography
  • Table 8 Compositions of high concentration anti-CD20 antibody (rituximab) formulations prepared as per Example-2

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Abstract

La présente invention concerne des formulations pharmaceutiques d'anticorps et de fragments de liaison à l'antigène contre CD20 humain et leur procédé de préparation. La composition de formulation divulguée stabilise l'anticorps anti-CD20 de concentrations plus faibles à plus élevées, ce qui la rend appropriée pour différents modes d'administration (sous-cutanée/intraveineuse).
PCT/IN2024/051485 2023-08-12 2024-08-11 Formulations pharmaceutiques d'anticorps anti-cd 20 et leurs procédés de préparation Pending WO2025037336A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060088523A1 (en) * 2004-10-20 2006-04-27 Genentech, Inc. Antibody formulations
US20090162352A1 (en) * 2007-12-21 2009-06-25 Michael Adler Antibody formulation
WO2011029892A2 (fr) * 2009-09-11 2011-03-17 F. Hoffmann-La Roche Ag Formulations pharmaceutiques hautement concentrées
WO2021068971A1 (fr) * 2019-10-12 2021-04-15 Bio-Thera Solutions, Ltd. Formulation d'anticorps anti-cd20 et utilisation d'anticorps anti-cd20 pour le traitement de maladies positives pour le cd20
WO2024023762A1 (fr) * 2022-07-27 2024-02-01 Kashiv Biosciences, Llc Composition liquide stable d'ocrélizumab

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060088523A1 (en) * 2004-10-20 2006-04-27 Genentech, Inc. Antibody formulations
US20090162352A1 (en) * 2007-12-21 2009-06-25 Michael Adler Antibody formulation
WO2011029892A2 (fr) * 2009-09-11 2011-03-17 F. Hoffmann-La Roche Ag Formulations pharmaceutiques hautement concentrées
WO2021068971A1 (fr) * 2019-10-12 2021-04-15 Bio-Thera Solutions, Ltd. Formulation d'anticorps anti-cd20 et utilisation d'anticorps anti-cd20 pour le traitement de maladies positives pour le cd20
WO2024023762A1 (fr) * 2022-07-27 2024-02-01 Kashiv Biosciences, Llc Composition liquide stable d'ocrélizumab

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