Self-Buffering Protein Formulations

Abstract

The invention herein described, provides, among other things, self-buffering protein formulations. Particularly, the invention provides self-buffering pharmaceutical protein formulations that are suitable for veterinary and human medical use. The self-buffering protein formulations are substantially free of other buffering agents, stably maintain pH for the extended time periods involved in the distribution and storage of pharmaceutical proteins for veterinary and human medical use. The invention further provides methods for designing, making, and using the formulation. In addition to other advantages, the formulations avoid the disadvantages associated with the buffering agents conventionally used in current formulations of proteins for pharmaceutical use. The invention in these and other respects can be productively applied to a wide variety of proteins and is particularly useful for making and using self-buffering formulations of pharmaceutical proteins for veterinary and medical use, especially, in particular, for the treatment of diseases in human subjects.

1 . A composition comprising a pharmaceutical protein, wherein at the pH of the composition, 21° C., one atmosphere, and equilibrium with ambient atmosphere, the protein has a buffer capacity per unit volume of at least that of approximately 4.0 mM sodium acetate buffer in pure water in the range of pH 5.0 to 4.0 or pH 5.0 to 5.5 under the same conditions, wherein further, exclusive of the buffer capacity of said protein, the buffer capacity per unit volume of the composition under the same conditions is no more than that of 2.0 mM sodium acetate buffer in pure water in the range of pH 5.0 to 4.0 or pH 5.0 to 5.5 under the same conditions, wherein the composition has been approved for pharmaceutical use by an authority legally empowered to grant such approval. 
     
     
         2 . A composition comprising a pharmaceutical protein, wherein at the pH of the composition, 21° C., one atmosphere, and equilibrium with ambient atmosphere, the protein has a buffer capacity per unit volume of at least 1.50 mEq/liter-pH unit, wherein further, exclusive thereof, the buffer capacity per unit volume of the composition is less than 0.5 mEq/liter-pH unit, wherein the composition has been approved for pharmaceutical use by an authority legally empowered to grant such approval. 
     
     
         3 . A composition according to  claim 1 , wherein the protein provides at least 80% of the buffer capacity of the composition. 
     
     
         4 . A composition according to  claim 3 , wherein the concentration of the protein is between approximately 20 and 400 mg/ml. 
     
     
         5 . A composition according to  claim 4 , wherein the pH maintained by the buffering action of the protein is between approximately 3.5 and 8.0. 
     
     
         6 . A composition according to  claim 5 , wherein the pH maintained by the buffering action of the protein is between approximately 4 and 6. 
     
     
         7 . A composition according to  claim 5 , further comprising one or more pharmaceutically acceptable salts, wherein the total salt concentration is less than 150 mM. 
     
     
         8 . A composition according to  claim 7 , further comprising one or more pharmaceutically acceptable salts, wherein the total salt concentration is less than 100 mM. 
     
     
         9 . A composition according to  claim 5 , further comprising one or more pharmaceutically acceptable polyols. 
     
     
         10 . A composition according to  claim 9 , wherein the polyol is one or more of sorbitol, mannitol, sucrose, trehalose, or glycerol. 
     
     
         11 . A composition according to  claim 5 , further comprising one or more pharmaceutically acceptable surfactants. 
     
     
         12 . A composition according to  claim 11 , wherein the surfactant is one or more of polysorbate 20, polysorbate 80, other fatty acid esters of sorbitan, polyethoxylates, and poloxamer 188. 
     
     
         13 . A composition according to  claim 9 , further comprising one or more pharmaceutically acceptable surfactants. 
     
     
         14 . A composition according to  claim 1 , further comprising one or more pharmaceutically acceptable: osmotic balancing agents; anti-oxidants; antibiotics; antimycotics; bulking agents; lyoprotectants; anti-foaming agents; chelating agents; preservatives; colorants; analgesics; or additional pharmaceutical agents. 
     
     
         15 . A composition according to  claim 5 , further comprising one or more pharmaceutically acceptable: osmotic balancing agents; anti-oxidants; antibiotics; antimycotics; bulking agents; lyoprotectants; anti-foaming agents; chelating agents; preservatives; colorants; analgesics; or additional pharmaceutical agents. 
     
     
         16 . A composition according to  claim 7 , further comprising one or more pharmaceutically acceptable: osmotic balancing agents; anti-oxidants; antibiotics; antimycotics; bulking agents; lyoprotectants; anti-foaming agents; chelating agents; preservatives; colorants; analgesics; or additional pharmaceutical agents. 
     
     
         17 . A composition according to  claim 1 , wherein the protein is or comprises: an antibody, Fab fragment, Fab 2  fragment, Fab 3  fragment, Fc fragment, scFv fragment, bis-scFv(s) fragment, minibody, diabody, triabody tetrabody, VhH domain, V-NAR domain, V H  domain, V L  domain, camel Ig, Ig NAR, receptibody, peptibody, or a variant or a derivative thereof or a protein related thereto, or a modification thereof. 
     
     
         18 . A composition according to  claim 17 , wherein the protein comprises an Fc fragment or a part thereof, or a variant or a derivative of an Fc fragment or a part thereof or a protein related to an Fc fragment or part thereof, or a modification of any thereof. 
     
     
         19 . A composition according to  claim 18 , wherein the protein further comprises a first binding moiety of a pair of cognate binding moieties. 
     
     
         20 . A composition according to  claim 1 , wherein the protein is selected from the group consisting of proteins that bind specifically to one or more CD proteins, HER receptor family proteins, cell adhesion molecules, growth factors, nerve growth factors, fibroblast growth factors, transforming growth factors (TGF), insulin-like growth factors, osteoinductive factors, insulins and insulin-related proteins, coagulation and coagulation-related proteins, colony stimulating factors (CSFs), other blood and serum proteins blood group antigens; receptors, receptor-associated proteins, growth hormone receptors, T-cell receptors; neurotrophic factors. neurotrophins, relaxins, interferons, interleukins, viral antigens, lipoproteins, integrins, rheumatoid factors, immunotoxins, surface membrane proteins, transport proteins, homing receptors, addressins, regulatory proteins, and immunoadhesins. 
     
     
         21 . A composition according to  claim 1 , wherein the protein is selected from the group consisting of: OPGL specific binding proteins, myostatin specific binding proteins, IL-4 receptor specific binding proteins, IL1-R1 specific binding proteins, Ang2 specific binding proteins, NGF-specific binding proteins, CD22 specific binding proteins, IGF-1 receptor specific binding proteins, B7RP-1 specific binding proteins, IFN gamma specific binding proteins, TALL-1 specific binding proteins, stem cell factors, Flt-3 ligands, and IL-17 receptors. 
     
     
         22 . A composition according to  claim 1 , wherein the protein is selected from the group consisting of proteins that bind specifically to one or more of: CD3, CD4, CD8, CD19, CD20, CD34; HER2, HER3, HER4, the EGF receptor; LFA-1, Mol, p150,95, VLA-4, ICAM-1, VCAM, alpha v/beta 3 integrin; vascular endothelial growth factor (“VEGF”); growth hormone, thyroid stimulating hormone, follicle stimulating hormone, luteinizing hormone, growth hormone releasing factor, parathyroid hormone, mullerian-inhibiting substance, human macrophage inflammatory protein (MIP-1-alpha), erythropoietin (EPO), NGF-beta, platelet-derived growth factor (PDGF), aFGF, bFGF, epidermal growth factor (EGF), TGF-alpha, TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta4, TGF-beta5, IGF-I, IGF-II, des(1-3)-IGF-I (brain IGF-I), insulin, insulin A-chain, insulin B-chain, proinsulin, insulin-like growth factor binding proteins;, such as, among others, factor VIII, tissue factor, von Willebrands factor, protein C, alpha-1-antitrypsin, plasminogen activators, such as urokinase and tissue plasminogen activator (“t-PA”), bombazine, thrombin, and thrombopoietin; M-CSF, GM-CSF, G-CSF, albumin, IgE, flk2/flt3 receptor, obesity (OB) receptor, bone-derived neurotrophic factor (BDNF), NT-3, NT-4, NT-5, NT-6); relaxin A-chain, relaxin B-chain, prorelaxin; interferon-alpha, -beta, and -gamma; IL-1 to IL-10; AIDS envelope viral antigen; calcitonin, glucagon, atrial natriuretic factor, lung surfactant, tumor necrosis factor-alpha and -beta, enkephalinase, RANTES, mouse gonadotropin-associated peptide, Dnase, inhibin, and activin; protein A or D, bone morphogenetic protein (BMP), superoxide dismutase, decay accelerating factor (DAF). 
     
     
         23 . A composition according to  claim 1 , wherein the protein is selected from the group consisting of: Actimmune (Interferon-gamma-1b), Activase (Alteplase), Aldurazme (Laronidase), Amevive (Alefacept), Avonex (Interferon beta-1a), BeneFIX (Nonacog alfa), Beromun (Tasonermin), Beatseron (Interferon-beta-1b), BEXXAR (Tositumomab), Tev-Tropin (Somatropin), Bioclate or RECOMBINATE (Recombinant), CEREZME (Imiglucerase), ENBREL (Etanercept), Eprex (epoetin alpha), EPOGEN/Procit (Epoetin alfa), FABRAZYME (Agalsidase beta), Fasturtec/Elitek ELITEK (Rasburicase), FORTEO (Teriparatide), GENOTROPIN (Somatropin), GlucaGen (Glucagon), Glucagon (Glucagon, rDNA origin), GONAL-F (follitropin alfa), KOGENATE FS (Octocog alfa), HERCEPTIN (Trastuzumab), HUMATROPE (SOMATROPIN), HUMIRA (Adalimumab), Insulin in Solution, INFERGEN® (Interferon alfacon-1), KINERET® (anakinra), Kogenate FS (Antihemophilic Factor), LEUKIN (SARGRAMOSTIM Recombinant human granulocyte-macrophage colony stimulating factor (rhuGM-CSF)), CAMPATH (Alemtuzumab), RITUXAN® (Rituximab), TNKase (Tenecteplase), MYLOTARG (gemtuzumab ozogamicin), NATRECOR (nesiritide), ARANESP (darbepoetin alfa), NEULASTA (pegfilgrastim), NEUMEGA (oprelvekin), NEUPOGEN (Filgrastim), NORDITROPIN CARTRIDGES (Somatropin), NOVOSEVEN (Eptacog alfa), NUTROPIN AQ (somatropin), Oncaspar (pegaspargase), ONTAK (denileukin diftitox), ORTHOCLONE OKT (muromonab-CD3), OVIDREL (choriogonadotropin alfa), PEGASYS (peginterferon alfa-2a), PROLEUKIN (Aldesleukin), PULMOZYME (domase alfa), Retavase (Reteplase), REBETRON Combination Therapy containing REBETOL® (Ribavirin) and INTRON® A (Interferon alfa-2b), REBIF (interferon beta-1a), REFACTO (Antihemophilic Factor), REFLUDAN (lepirudin), REMICADE (infliximab), REOPRO (abciximab)ROFERON®-A (Interferon alfa-2a), SIMULECT (baasiliximab), SOMAVERT (Pegivisomant), SYNAGIS® (palivizumab), Stemben (Ancestim, Stem cell factor), THYROGEN, INTRON® A (Interferon alfa-2b), PEG-INTRON® (Peginterferon alfa-2b), XIGRIS® (Drotrecogin alfa activated), XOLAIR® (Omalizumab), ZENAPAX® (daclizumab), and ZEVALIN® (Ibritumomab Tiuxetan). 
     
     
         24 . A composition according to  claim 1 , wherein the protein is Ab-hOPGL or a fragment thereof, or a variant or derivative of Ab-hOPGL or of a fragment thereof, or an Ab-hOPGL related protein or fragment thereof, or a modification of any thereof. 
     
     
         25 . A composition according to  claim 1 , wherein the protein is Ab-hOPGL. 
     
     
         26 . A composition according to  claim 1 , wherein the protein is Ab-hIL4R or a fragment thereof, or a variant or derivative of Ab-hIL4R or of a fragment thereof, or an Ab-hIL4R related protein or fragment thereof, or a modification of any thereof. 
     
     
         27 . A composition according to  claim 1 , wherein the protein is Ab-hIL4R. 
     
     
         28 . A composition according to  claim 1 , wherein the protein is Ab-hB7RP1 or a fragment thereof, or a variant or derivative of Ab-hB7RP1 or of a fragment thereof, or an Ab-hB7RP1 related protein or fragment thereof, or a modification of any thereof. 
     
     
         29 . A composition according to  claim 1 , wherein the protein is Ab-hB7RP1. 
     
     
         30 . A lyophilate which upon reconstitution provides a composition according to  claim 1 . 
     
     
         31 . A kit comprising in one or more containers a composition according to  claim 1 , and instructions regarding the use thereof. 
     
     
         32 . A kit comprising in one or more containers a lyophilate according to  claim 31 , and instructions regarding the use thereof. 
     
     
         33 . A method for treating a subject, comprising administering to a subject in an amount and by a route effective for treatment, a composition according to  claim 1 . 
     
     
         34 . A process for preparing a composition according to  claim 1 , comprising removing residual buffer using a counter ion. 
     
     
         35 . A process for preparing a composition according to  claim 34 , comprising removing residual buffer using any one or more of the following in the presence of a counter ion: size exclusion chromatography, dialysis, and/or tangential flow filtration. 
     
     
         36 . A process for preparing a composition according to  claim 35 , comprising removing residual buffer using ion exchange chromatography. 
     
     
         37 . A process for preparing a composition according to  claim 1 , comprising removing residual buffer by diafiltration against a bufferless solution having a pH below the desired pH. 
     
     
         38 . A process for preparing a composition according to  claim 37 , wherein following diafiltration the pH is adjusted to a desired pH by addition of dilute acid and/or dilute base. 
     
     
         39 . A lyophilate which upon reconstitution provides a composition according to  claim 20 . 
     
     
         40 . A kit comprising in one or more containers a composition according to  claim 20  and instructions regarding the use thereof. 
     
     
         41 . A kit comprising in one or more containers a lyophilate according to  claim 40 , and instructions regarding the use thereof. 
     
     
         42 . A method for treating a subject, comprising administering to a subject in an amount and by a route effective for treatment, a composition according to  claim 20 . 
     
     
         43 . A process for preparing a composition according to  claim 20 , comprising removing residual buffer using a counter ion. 
     
     
         44 . A process for preparing a composition according to  claim 43 , comprising removing residual buffer using any one or more of the following in the presence of a counter ion: size exclusion chromatography, dialysis, and/or tangential flow filtration. 
     
     
         45 . A process for preparing a composition according to  claim 43 , comprising removing residual buffer using ion exchange chromatography. 
     
     
         46 . A process for preparing a composition according to  claim 20 , comprising removing residual buffer by diafiltration against a bufferless solution having a pH below the desired pH. 
     
     
         47 . A process for preparing a composition according to  claim 46 , wherein following diafiltration the pH is adjusted to a desired pH by addition of dilute acid and/or dilute base. 
     
     
         48 . A lyophilate which upon reconstitution provides a composition according to  claim 21 . 
     
     
         49 . A kit comprising in one or more containers a composition according  claim 21  and instructions regarding the use thereof. 
     
     
         50 . A kit comprising in one or more containers a lyophilate according to  claim 49 , and instructions regarding the use thereof. 
     
     
         51 . A method for treating a subject, comprising administering to a subject in an amount and by a route effective for treatment, a composition according to  claim 21 . 
     
     
         52 . A process for preparing a composition according to  claim 21 , comprising removing residual buffer using a counter ion. 
     
     
         53 . A process for preparing a composition according to  claim 52 , comprising removing residual buffer using any one or more of the following in the presence of a counter ion: size exclusion chromatography, dialysis, and/or tangential flow filtration. 
     
     
         54 . A process for preparing a composition according to  claim 52 , comprising removing residual buffer using ion exchange chromatography. 
     
     
         55 . A process for preparing a composition according to  claim 21 , comprising removing residual buffer by diafiltration against a bufferless solution having a pH below the desired pH. 
     
     
         56 . A process for preparing a composition according to  claim 55 , wherein following diafiltration the pH is adjusted to a desired pH by addition of dilute acid and/or dilute base. 
     
     
         57 . A lyophilate which upon reconstitution provides a composition according to  claim 23 . 
     
     
         58 . A kit comprising in one or more containers a composition according  claim 23  and instructions regarding the use thereof. 
     
     
         59 . A kit comprising in one or more containers a lyophilate according to  claim 57 , and instructions regarding the use thereof. 
     
     
         60 . A method for treating a subject, comprising administering to a subject in an amount and by a route effective for treatment, a composition according to  claim 23 . 
     
     
         61 . A process for preparing a composition according to  claim 23 , comprising removing residual buffer using a counter ion. 
     
     
         62 . A process for preparing a composition according to  claim 61 , comprising removing residual buffer using any one or more of the following in the presence of a counter ion: size exclusion chromatography, dialysis, and/or tangential flow filtration. 
     
     
         63 . A process for preparing a composition according to  claim 61 , comprising removing residual buffer using ion exchange chromatography. 
     
     
         64 . A process for preparing a composition according to  claim 21 , comprising removing residual buffer by diafiltration against a bufferless solution having a pH below the desired pH. 
     
     
         65 . A process for preparing a composition according to  claim 62 , wherein following diafiltration the pH is adjusted to a desired pH by addition of dilute acid and/or dilute base.