MX2008008021A - Protein formulations with reduced viscosity and uses thereof - Google Patents

Abstract

Protein formulations and methods for reducing the viscosity of a protein formulation are provided. The method for reducing the viscosity of a protein formulation comprises adding a viscosity reducing agent, such as calcium chloride or magnesium chloride to the protein formulation.

Description

PROTEIN FORMULATIONS WITH REDUCED VISCOSITY AND ITS USES FIELD OF THE INVENTION Field refers to protein formulations and, more particularly to protein formulations with reduced viscosity. BACKGROUND OF THE INVENTION It is estimated that more than 371 new medic based on biotechnology are in the industrial path. Biotechnology-based medic include therapeutic proteins such as enzymes, soluble receptors, ligands, blood proteins and monoclonal antibodies. Protein-based therapy, especially monoclonal antibody-based therapy, has become an important method to treat diseases such as cancer, allergic diseases, asthma and organ transplantation. At the end of 2003, 14 antibody-based therapies were approved through the administration of food and drugs to treat different human diseases. Antibody-based therapy is usually administered over regular doses and requires several mg / kg dosages per injection. Subcutaneous injection is a typical route of administration of these therapies. Due to the small volumes used for subcutaneous injection (usually 1.0 ml-1.2 ml), for Ref. 194178 With high-dose antibody therapies, this route of administration requires protein formulations with high concentrations (eg, 50 mg / ml-300 mg / ml). The high protein concentrations have challenges related to the physical and chemical stability of the protein, and the difficulty with the manufacture, storage and distribution of the protein formulation. One problem is the tendency of proteins to form particles during processing and / or storage, which makes handling difficult during further processing. To try to obviate this problem, the surfactants and / or sugars have been added to the protein formulations. Although surfactants and sugars can reduce the degree of particle formation of the protein, they do not cover another problem associated with the handling and administration of concentrated protein formulations, i.e., increased viscosity. In fact, sugars can improve intermolecular interactions within a protein or between proteins and increase the viscosity of the protein formulation. The increased viscosity of the protein formulations has negative ramifications from processing by distributing the drug to the patient. Accordingly, there is a need in the art to develop protein formulations with relatively high concentrations with suitably low viscosities that are suitable for manufacturing, storage, and administration. BRIEF DESCRIPTION OF THE INVENTION The present application relates to protein formulations having reduced viscosity compared to corresponding protein formulation that does not include a viscosity reducing agent in a suitable concentration, and methods for making the protein formulations having reduced viscosity (formulations of reduced viscosity). In one aspect, the invention relates to methods for reducing the viscosity of a protein formulation through the addition of a viscosity reducing agent or a protein formulation, thereby reducing the viscosity of the protein formulation compared to a protein formulation lacking viscosity reducing agent. In one embodiment, the method involves determining the viscosity of a protein formulation prior to the addition of a viscosity reducing agent. In another embodiment, the method involves determining the viscosity of a protein formulation after the addition of a viscosity reducing agent. In yet another embodiment, the method involves determining the viscosity of a protein formulation before and after the addition of the viscosity reducing agent. In certain modalities, the viscosity reducing agent reduces the viscosity of the protein formation by at least 5% compared to the viscosity of the formulation formulated without the viscosity reducing agent. In some embodiments, the viscosity reducing agent is calcium chloride or magnesium chloride. The viscosity reducing agent is added at low concentrations so as not to have a negative impact on the protein formulation. The viscosity reducing agent is generally added to the protein formulation at a final concentration of between about 1 mM and about 50 mM. In some embodiments, the viscosity reducing agent is added to a protein formulation at a final concentration of between about 5 mM and about 25 mM. In certain embodiments, the viscosity reducing agent is added to a protein formulation at a final concentration of between about 1 mM and about 20 mM. In certain embodiments, the viscosity reducing agent is added to a protein formulation at a final concentration of between about 0.5 mM and 14 mM. In another embodiment, the protein is an antibody, an Ig fusion protein, a receptor, a ligand, a transcription factor, an enzyme, or a biologically active fragment thereof. In some embodiments, the protein is an anti-myostatin antibody, an antibody anti-IL-12, or an anti-IL-13 antibody. In another aspect, the invention relates to a protein formulation with reduced viscosity. The protein formulation with reduced viscosity includes a protein, a viscosity reducing agent, and a buffer. In some embodiments, the viscosity reducing agent is calcium chloride or magnesium chloride. The viscosity reducing agent generally adds a protein formulation at a general concentration of between about 1 mM and about 50 mM. In some embodiments, the viscosity reducing agent is added to the protein formulation at a final concentration of between about 5 mM and about 25 mM. In certain embodiments, the viscosity reducing agent is added to the protein formulation at a final concentration of between about 0.5 mM and about 15 mM. In certain other embodiments, the viscosity reducing agent is added to a protein formulation at a final concentration of between 0.5 mM and 14 mM. When the viscosity reducing agent is added to a protein formulation at a concentration of between about 0.5 mM to about 50 mM, sodium chloride or sodium biphosphate is not used as viscosity reducing agents. The pH of the protein formulation is generally between about 5.5 and about 6.5. In certain embodiments, the protein is an antibody, an Ig fusion protein, a receptor, a ligand, a transcription factor, an enzyme, or a biologically active fragment thereof. In certain embodiments, the protein formulation is provided as kits. The kits may include instructions for the use of the protein formulation. In certain embodiments, the reduced viscosity protein formulation is a reduced viscosity anti-myostatin antibody formulation. In one embodiment, the anti-myostatin antibody is a monoclonal antibody. In another embodiment, the anti-myostatin antibody is a humanized monoclonal antibody (e.g., a partially humanized or fully humanized monoclonal antibody). In certain embodiments, the anti-myostatin antibody is MYO-022, MYO-028 or MYO-029. Anti-myostatin antibodies are generally used at a concentration of between about 25 mg / ml to about 400 mg / ml. The viscosity reducing agent is generally added to an anti-myostatin antibody formulation with reduced viscosity at a final concentration of between about 1 mM and about 50 mM. In some embodiments, the viscosity reducing agent is added to an anti-myostatin antibody at a final concentration of between about 5 mM and about 25 mM. In certain embodiments, the viscosity reducing agent is addition to the anti-myostatin antibody formulation at a final concentration of between about 1 mM and about 15 mM. In certain embodiments, the viscosity reducing agent is added to an anti-myostatin antibody formulation at a final concentration of between about 0 mM and 14 mM. When the viscosity reducing agent is added to an anti-myostatin antibody formulation at a concentration of between about 0.5 mM to about 50 mM, sodium chloride or sodium biphosphate is not used as viscosity reducing agents. Anti-myostatin antibody formulations of reduced viscosity generally have a pH of between about 5.5 and about 6.5. In one embodiment, histidine is used to regulate the pH of a reduced viscosity myostatin antibody formulation. A myostatin antibody formulation of reduced viscosity may also include one or more cryoprotectants, one or more surfactants, one or more anti-oxidants, or a combination thereof. In some embodiments, the anti-myostatin formulation with reduced viscosity is a reconstituted formulation. The myostatin antibodies can be formulated as described herein as pharmaceutical compositions and are used to treat disorders such as, but not limited to, muscular dystrophy, sarcopenia, cachexia, and type II diabetes. In certain modalities, a Formulation of reduced viscosity anti-myostatin antibody is provided as a kit. The kits may include instructions for the use of the antibody formulation. In certain embodiments, the reduced viscosity protein formulation is an anti-IL-12 antibody formulation of reduced viscosity. In one embodiment, the anti-IL-12 antibody is a monoclonal antibody. In another embodiment, the anti-IL-12 antibody is a humanized monoclonal antibody (e.g., a partially humanized or fully humanized monoclonal antibody). In certain embodiments, the anti-IL-12 antibody is J695. Anti-IL-12 antibodies are generally used in a formulation or a concentration of between about 25 mg / ml to about 400 mg / ml. A viscosity reducing agent is generally added to an anti-IL-12 antibody formulation at a final concentration of between about 1 mM and about 50 mM. In some embodiments, the viscosity reducing agent is added to an anti-IL-12 antibody formulation at a final concentration of between about 5 mM and about 25 mM. In certain embodiments, the reducing agent is added to an anti-IL-12 antibody formulation at a final concentration of between about 1 mM and about 15 mM. In certain other embodiments, the viscosity reducing agent is added to an anti- body antibody formulation.

IL-12 at a final concentration of between about 0.5 mM and about 14 M. When the viscosity reducing agent is added to an anti-IL-12 antibody formulation at a concentration of between about 0.5 mM to about 50 mM, no uses sodium chloride and sodium bisphosphate as viscosity reducing agents. Formulations of anti-IL-12 antibody with reduced viscosity generally have a pH of between about 5.5 and about 6.5. In certain embodiments, histidine is used as a buffer in an IL-12 antibody formulation with reduced viscosity. Formulations of anti-IL-12 antibody with reduced viscosity may also include one or more cryoprotectants, one or more surfactants, one or more anti-oxidants, or combinations thereof. In certain embodiments, the reduced viscosity anti-IL-12 formulation is a reconstituted formulation. Anti-IL-12 antibodies can also be formulated as described herein for use as pharmaceutical compositions and used to treat disorders such as, but not limited to, rheumatoid arthritis, Crohn's disease, psoriasis, and psoriatic arthritis. In certain embodiments, the formulation of the anti-IL-12 antibody of reduced viscosity is provided as part of a kit. The kits may include instructions for use in the formulation of the anti-IL-12 antibody.

In certain embodiments, the reduced viscosity protein formulation is a reduced viscosity anti-IL-13 antibody formulation. In one embodiment, the anti-IL-13 antibody is a monoclonal antibody. In another embodiment, the anti-IL-13 antibody is a humanized monoclonal antibody (e.g., a partially humanized or fully humanized monoclonal antibody). In certain embodiments, the anti-IL-13 antibody is IMA-638. Anti-IL-13 antibodies are generally used in a formulation or a concentration of between about 25 mg / ml to about 400 mg / ml. A viscosity reducing agent is generally added to an anti-IL-13 antibody formulation at a final concentration of between about 1 mM and about 50 mM. In some embodiments, the viscosity reducing agent is added to an anti-IL-13 antibody formulation at a final concentration of between about 5 mM and about 25 mM. In certain embodiments, the reducing agent is added to an anti-IL-13 antibody formulation at a final concentration of between about 1 mM and about 15 mM. In certain other embodiments, the viscosity reducing agent is added to an anti-IL-13 antibody formulation at a final concentration of between about 0.5 mM and about 14 mM. When the viscosity reducing agent is added to an anti- body antibody formulation, IL-13 at a concentration of between about 0.5 mM to about 50 mM, sodium chloride and sodium bisphosphate are not used as viscosity reducing agents. Formulations of anti-IL-13 antibody with reduced viscosity generally have a pH of between about 5.5 and about 6.5. In one embodiment, histidine is used as a buffer in an IL-13 antibody formulation with reduced viscosity. Formulations of anti-IL-13 antibody with reduced viscosity may also include one or more cryoprotectants, one or more surfactants, one or more anti-oxidants, or combinations thereof. In certain embodiments, the anti-IL-13 formulation of reduced viscosity is a reconstituted formulation. Anti-IL-13 antibodies can be formulated in a reduced viscosity formulation as pharmaceutical compositions and used to treat disorders such as, but not limited to, respiratory disorders (eg, asthma); atopic disorders (for example, allergic rhinitis); inflammatory and / or autoimmune conditions of the skin (eg, atopic dermatitis), gastrointestinal organs (for example, inflammatory bowel diseases (IBD)), as well as fibrotic and carcinogenic disorders. In certain embodiments, the formulation of the anti-IL-13 antibody of reduced viscosity is provided as a kit. The kits may include instructions for use in formulating the anti-IL-13 antibody of reduced viscosity. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used in practice or to test the present invention, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. In addition, the materials, methods and examples are illustrative only and are not intended to be limiting. Other features and advantages of the invention will be apparent from the detailed description, the figures and the claims. BRIEF DESCRIPTION OF THE FIGURES Figure 1 is a graph describing the results of the conducted experiments to determine the effect of increasing the concentrations of various salts in the viscosity of the anti-myostatin antibody formulation (MYO-029). Figure 2 is a bar graph that describes the results of the experiments conducted to determine the effect of increasing the concentrations of calcium chloride in the viscosity of an anti-myostatin antibody formulation (MYO-028). Figure 3 is a graph describing the results of the conducted experiments to determine the effect of increasing the calcium chloride concentrations in the viscosity of an anti-IL-13 antibody formulation (IMA-638). Figure 4A is a bar graph describing the results of the experiments conducted to test the effect of the degradation induced by freeze-thaw of the anti-myostatin antibody (MYO-029) in the presence or absence of calcium chloride. The degradation was evaluated through the recovery of the protein as determined by measuring the absorbance at 280 nm. Figure 4B is a bar graph describing the results of experiments conducted to test the effect of freeze-thaw induced degradation of anti-myostatin antibody (MYO-029) in the presence or absence of calcium chloride. The degradation was evaluated through the percentage of the formation of high molecular weight species (% HMW) determined by high performance liquid chromatography-size exclusion (SEC-HPLC). Figure 5A is a bar graph describing the results of the experiments conducted to test the effect of the presence or absence (control) of calcium chloride on the liquid stability of the anti-myostatin antibody (MYO-029) subjected to storage at 40 ° C for up to seven days. The liquid stability of MYO-029 antibody was determined by measuring the absorbance at 280 nm. Figure 5B is a bar graph describing the results of the experiments conducted to test the effect of the presence or absence (control) of calcium chloride on the liquid stability of the anti-myostatin antibody (MYO-029) subjected to storage at 40 ° C for up to seven days. The liquid stability of antibody MYO-029 was determined by measuring the HMW formation determined by SEC-HPLC. Figure 6 is a representation of the amino acid sequence of the heavy chain antibody MYO-028 (SEQ ID NO: 1) and the light chain (SEQ ID NO: 2). Figure 7 is a representation of the amino acid sequence of the heavy chain MYO-029 antibody (SEQ ID NO: 3) and the light chain (SEQ ID NO: 4). Figure 8 is a depiction of the amino acid sequence of heavy chain (SEQ ID NO: 5) J695 and light chain antibody (SEQ ID NO: 6). Figure 9 is a representation of the sequence of amino acid of heavy chain (SEQ ID NO: 7) and light chain IMA-638 antibody (SEQ ID NO: 8). The last amino acid residue encoded by the heavy chain DNA sequence, Lys448, is observed in secreted, mature form of IMA-638 only small amounts and presumably removed from the monoclonal antibody volume during intracellular processing via CHO cellular proteases . Accordingly, the carboxy terminus of heavy chain IMA-638 is Gly447. The lysine processing of the term carboxy has been observed in recombinant antibodies and plasma derivatives and does not seem to have an impact on its functions. DETAILED DESCRIPTION OF THE INVENTION The viscosity of the protein formulation has implications for stability, processing, storage, and, for those used as drugs, drug distribution of the protein formulation to a patient. The implications include, but are not limited to, the concentration of pH buffer exchange through ultrafiltration and diafiltration (the flow through the membrane can decrease with an increased viscosity thereby resulting in longer processing times), sterile filtration (it took longer to filter sterile viscous solutions, and in some cases a very viscous solution will not pass through the membranes with very small pores, eg, 0.22 μm membranes), handling of samples (eg, difficulty with pipette measurement and the ability to extract it in a syringe), recovery of bottle storage after reconstitution, stability, and passage through needles for subcutaneous or intramuscular administration. Methods for reducing the viscosity of a protein formulation that has been identified are provided herein. The methods are suitable for preparing protein formulations that have a reduced viscosity ("reduced viscosity formulations" or "reduced viscosity protein formulations"). These reduced viscosity protein formulations include a protein of interest and a viscosity reducing agent. Methods to Reduce the Viscosity of a Protein Formulation The term "viscosity" as used herein may be "kinematic viscosity" or "absolute viscosity". "Kinematic viscosity" is a measurement of the resistive flow of the fluid under the influence of gravity. When two fluids of equal volume are placed in identical capillary viscometers and allowed to flow through gravity, the viscous fluid takes more time than the viscous fluid to flow through the capillary. If one of the fluids takes 100 seconds to complete its flow and another fluid takes 200 seconds, the second fluid is twice as viscous as the first on a scale of kinematic viscosity. "Absolute viscosity" sometimes referred to as "dynamic viscosity" or "simple viscosity", is a product of kinematic viscosity and fluid density. The dimension of the kinematic viscosity is L2 / T where L is a length and T is a time. Commonly, the kinematic viscosity is expressed in centistokes (cSt). The SI unit of the kinematic viscosity is mm2 / s, where is 1 cSt. The absolute viscosity is expressed in a unit of centipoise (cP). The SI unit of the absolute viscosity is the milliPascal-second (mPa-s), where 1 CP = 1 mPa-s. The viscosity of a protein formulation can be reduced through the addition of a viscosity reducing agent of the formulation. In some cases, the viscosity reducing agent is added at a relatively low concentration. The viscosity of a formulation comprising a viscosity reducing agent is reduced compared to the viscosity of a formulation lacking viscosity reducing agent. When the addition of the viscosity reducing agent results in the decrease in the viscosity of the formulation compared to a corresponding formulation that does not include the viscosity reducing agent or compared to a formulation that does not include the viscosity reducing agent at a concentration selected, the formulation containing the viscosity reducing agent (eg, at a selected concentration), the formulation is a formulation with reduced viscosity. In certain formulations of reduced viscosity, the viscosity reducing agent generally reduces the viscosity of the protein formulation by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, and about 90% compared to the viscosity of a protein formulation without, or containing, lower amounts of the viscosity reducing agent. In some cases, the viscosity reducing agent reduces the viscosity of a protein formulation by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, and at least 90% compared to the viscosity of a protein formulation, without or containing lower amounts of the viscosity reducing agent. In certain embodiments, the viscosity of the protein formulation is measured before the addition of the viscosity reducing agent. In other embodiments, the viscosity of the protein formulation is measured after the addition of the viscosity reducing agent. Measurements can be done in hours (for example, 1-23 hours), days (for example, 1-10 days), weeks (for example, 1-5 weeks), or months (for example, 1-12 months) , or years (eg, 1-2 years, 1-3 years) after the addition of the viscosity reducing agent to a protein formulation. In yet other embodiments, the viscosity of the protein formulation is measured before and after the addition of the viscosity reducing agent. Methods for measuring viscosity are well known in the art and include, for example, using a capillary viscometer, or a rheometer with a cone plate. In one embodiment, the viscosity reducing agent is a salt such as calcium chloride, magnesium chloride, sodium phosphate or arginine hydrochloride. In the method described herein, the viscosity reducing agent is added to the formulation of the protein at a final concentration of between about 0.5 mM and about 100 mM. In one embodiment, the viscosity reducing agent is added to the protein formulation at a final concentration of between about 5 mM and about 20 mM. In another embodiment, the viscosity reducing agent is added to the protein formulation at a final concentration of between approximately 0.5 mM and 14 mM. In certain embodiments, the viscosity reducing agent is added to the formulation of the protein at a final concentration of between about 0.5 mM and not more than 20 mM, or 19 mM, or 18 mM, or 17 mM, or 16 mM, or 15 mM, or 14 mM, or 13 mM, or 12 mM, or 11 mM, or 10 mM. In general, when the viscosity reducing agent is added to the protein formulation at a final concentration of between about 0.5 mM and about 25 mM, the viscosity reducing agent is calcium chloride or magnesium chloride but not sodium chloride, or sodium biphosphate. In certain embodiments, the viscosity reducing agent is added at low concentrations so as not to negatively impact the protein formulation. For example, at concentrations of calcium chloride or magnesium chloride of 20 mM or greater, the proteins can form a gel at low storage temperatures (eg, 2-8 ° C). Accordingly, a concentration of a viscosity reducing agent is generally selected for which the viscosity is reduced to the expected storage temperature of the reduced viscosity formulation. Formulations The composition of a protein formulation of reduced viscosity is determined through the consideration of several factors. These factors include, but are not limited to: the nature of the protein (for example, receptor, antibody, Ig fusion proteins, enzymes); the concentration of the protein; the desired pH range; how the protein formulation is to be stored (eg, temperature); the period of time over which the protein formulation will be stored; and how the formulation is to be administered to a patient. The selection of an appropriate viscosity reducing agent is made based, in part, on the requirements for the protein in the formulation. Proteins The protein of interest to be formulated includes, but is not limited to, proteins such as, myostatin / GDF-8; interleukins (ILs), for example, IL-1 to IL-15; growth hormones such as human growth hormone and bovine growth hormone; growth hormone release factor; parathyroid hormone; thyroid stimulating hormone; uricase, bicunin, bilirubin oxidase, subtilisin; α-1-antitrypsin lipoproteins; insulin chain A; insulin chain B proinsulin; follicle stimulating hormone; calcitonin leutinizing hormone; glucagon; Vlla Factor; Factor VIII, Factor VIIIC; Factor IX; tissue factor, von Willebrand factor; anti-coagulation factors such as protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator; such as urokinase or Tissue type plasminogen activator (t-PA); bombazine; thrombin; plasmite; miniplasmin; microplasmin; tumor necrosis factor a and ß; enkephalinase; RANTES (Regulated on Normally Expressed and Secreted T Cell Activation); inflammatory protein of human macrophage (MIP-1-a); serum albumin such as human serum albumin; Mulerian inhibitory substance; relaxin chain A; Relaxin B chain; pro-relaxin; peptide associated with mouse gonadotropin; DNase; inhibin; activita; vascular endothelial growth factor (VEGF, for its acronym in English); placenta growth factor (PIFG); receptors for hormones or growth factors; an integrin; protein A or protein D; rheumatoid factors; a neurotrophic factor such as bone-derived neurotrophic factor (BDNF), neutrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6), or a factor of nerve growth such as NGF-β; platelet-derived growth factor (PDGF); fibroblast growth factor such as FGF and bFGF; epidermal growth factor (EGF); Transforming growth factor (TGF) such as TGF-α and TGF-β, including TGF-β1, TGF-β2, TGF-β3, TGF-β4, or TGF-β5; insulin-type growth factor I and II (IGF-I and IGF-II); des (1-3) -IGF-I (brain IGF-I); insulin-like growth factor binding proteins; proteins CDs such as: CD2, CD3, CD4, CD8, CD9, CD19, CD20, CD22, CD28, CD34, and CD45; erythropoietin (EPO); thrombopoietin (TPO); osteoinductive factors; immunotoxins; a bone morphogenetic protein (BMP); an interferon such as interferon a, β and β; a colony stimulating factor (CSF), for example, M-CSF, GM-CSF, and G-CSF; superoxide dismutase; T cell receptors; members of the HER receptor family such as the EGF receptor, HER2, HER3 or HER4; cell adhesion molecules such as LFA-1, VLA-4, ICAM-1, and VCAM; IgE; blood group antigens; the flk2 / flt3 receptor; Obesity receptor (OB); decay acceleration factor (DAF, for its acronym in English); a viral antigen such as HIV gag, env, pol, tat, or rev proteins; habitat recipients; adresinas; immunoadhesins; and biologically active fragments or variants of any of the aforementioned polypeptides. In some formulations, more than one type of protein or fragment is included in the formulation. The term "biologically active fragment" means a fragment of a protein that retains at least one of the functions of the protein from which it is derived. A biologically active fragment of an antibody that includes an antibody antigen binding fragment; a biologically active fragment of a receptor includes a fragment of the receptor that still binds to its ligand; a A biologically active fragment of a ligand includes that portion of ligand that can still bind to its receptor, and a biologically active fragment of an enzyme includes that portion of the enzyme that can still catalyze a reaction catalyzed by the full-length enzyme. In one embodiment, a biologically active fragment retains at least 5% of the function of the protein from which it is derived. The function of a protein can be tested by methods known in the art (e.g., testing antibody-antigen interactions, testing ligand-receptor interactions, testing enzymatic activity, testing transcription activity, or testing the interactions of DNA-protein). In some cases, the fragment is a therapeutically useful fragment which may, for example, retain certain characteristics of the protein from which it is derived (e.g., binding to a specific ligand) but does not cause the cellular response produced by the protein from which it is derived. In certain embodiments, the protein to be formulated is an antibody. The antibody can be one that binds to one of the aforementioned proteins. The term "antibody" as used herein, includes polyclonal antibodies, monoclonal antibodies, antibody compositions with polyepitope specificities, biospecific antibodies, diabodies, or other purified preparations of antibodies and recombinant antibodies. The antibodies can be whole antibodies, for example, any isotype (IgG, IgA, IgE, IgM, etc.), or fragment thereof, which binds to the antigen of interest. In a specific example of an antibody used in the present invention, the antibody to be formulated is an antibody having the IgG isotype. The antibodies can be fragmented using conventional or other techniques and the fragments classified for binding to an antigen of interest. Generally, an antibody fragment comprises the antigen binding and / or the variable reaction of an intact antibody. Thus, the term "antibody fragment" includes segments of proteolytically divided or recombinantly prepared portions of an antibody molecule that can selectively bind to a selected protein. Non-limiting examples of the proteolytic and / or recombinant fragments include Fab, F (ab ') 2, Fab', Fv, and a single chain antibody (scFv) containing a V [L] and / or V [H] domain ] linked to the peptide linker. The scFvs can be linked covalently or non-covalently to form antibodies having two or more binding sites. In some embodiments, the antibody is a humanized monoclonal antibody. The term "humanized monoclonal antibody" as used herein, is a monoclonal antibody of a non-human origin (receptor) that has been altered to contain at least one or more amino acid residues found in the equivalent human monoclonal antibody (donor). A "fully humanized monoclonal antibody" is a monoclonal antibody from a non-human source that has been altered to contain all amino acid residues found in the antigen binding region of the human equivalent monoclonal antibody. Humanized antibodies may also comprise residues that are found neither in the recipient antibody nor the donor antibody. These modifications can also be made to refine and optimize the functionality of the antibody. A humanized antibody can optionally also comprise at least a portion of a human immunoglobulin constant region (Fc). In certain embodiments, the antibody used in a reduced viscosity formulation is an anti-myostatin antibody (e.g., MYO-022, MYO-028 (Figure 6), MYO-029 (Figure 7)). The antibodies MYO-022, MYO-028, and MIO-029 are described in the patent application of E. U. A. No. 10 / 688,925 (Pub. No. 2004/0142382), which is incorporated herein by reference. In other embodiments, the antibody is an IL-12 antibody (e.g., J695 (Figure 8)). The J695 antibody is described in the E. U.A. No. 6,914,128, which is incorporated herein by reference. In yet another embodiment, the antibody is an anti-IL-13 antibody (by example, IMA-638 (Figure 9), CAT-354). Anti-IL-13 antibodies are described in the E patent application. U. A. No. 11 / 149,309, which is incorporated herein by reference. In some embodiments, the protein to be formulated is a fusion protein. In one embodiment, the fusion protein is an immunoglobulin (Ig) fusion protein. In a specific embodiment, the fusion protein comprises the IgG heavy chain constant region. In another embodiment, the fusion protein comprises an amino acid sequence corresponding to the connection, to the CH2 and CH3 regions of the human immunoglobulin C? L. Examples of Ig fusion proteins include CTLA4 Ig and VCAM2D-IgG. Methods for preparing fusion proteins are known in the art (for example, U.S. Patent Nos. 6,887,471 and 6,482,409). In certain embodiments, the protein to be formulated is a protein that does not include a Factor VII polypeptide, or an anti-IgE antibody. A reduced viscosity formulation may contain more than one protein as necessary for the treatment of a particular disorder. The additional protein (s) typically has activities complementary to other proteins in the formulation, and does not adversely affect the other protein (s) in the formulation. For example, it may be desirable to provide a single formulation containing two or more antibodies that bind to myostatin; two or more antibodies that bind to IL-12; or two or more antibodies that bind to IL-13. In addition, a protein formulation may also contain non-protein substances that are used in the final utility of the reduced viscosity protein formulation. For example, sucrose can be added to improve the stability and solubility of the protein in solution; and histidine may be added to provide the appropriate buffering capacity. The additional substances can be part of a protein formulation before the addition of a viscosity reducing agent or the addition in the process to make a formulation of reduced viscosity. In certain embodiments, the protein to be formulated is essentially pure and / or essentially homogeneous (ie, essentially free of contaminating proteins, etc.) before its use in the formulation. The term "essentially pure" protein means a composition comprising at least 90% by weight of a selected protein fraction, for example, at least about 95% by weight of the selected protein fraction. The term "essentially homogeneous" protein means a composition comprising at least 99% by weight of a selected protein fraction, excluding the mass of several stabilizers and water in solution.

Concentration of the Protein in a Low Vi scosity Formulation The concentration of the protein in a reduced viscosity formulation depends on the final use of the formulation. The protein concentrations in the formulations described herein are generally between about 10 mg / ml and about 300 mg / ml, for example, between about 10 mg / ml and about 100 mg / ml, about 25 mg / ml and about 100 mg / ml. mg / ml, approximately 50 mg / ml and approximately 100 mg / ml, approximately 75 mg / ml and approximately 100 mg / ml, approximately 100 mg / ml and approximately 200 mg / ml, approximately 125 mg / ml and approximately 200 mg / ml ml, approximately 150 mg / ml and approximately 200 mg / ml, approximately 200 mg / ml and approximately 300 mg / ml, and approximately 250 mg / ml and approximately 300 mg / ml. For example, protein concentrations in the formulation described herein can be between 10 mg / ml and 300 mg / ml, for example, between 10 mg / ml and 100 mg / ml, between 25 mg / ml and 100 mg / ml. ml, between 50 mg / ml and 100 mg / ml, between 75 mg / ml and 100 mg / ml, between 100 mg / ml and 200 mg / ml, between 125 mg / ml and 200 mg / ml, between 150 mg / ml ml and 200 mg / ml, between 200 mg / ml and 300 mg / ml, and between 250 mg / ml and 300 mg / ml. The term "between" is intended to be inclusive of the minimum and maximum concentrations.

Reduced viscosity protein formulations can be used for therapeutic purposes. Accordingly, the concentration of the protein in a formulation used by a therapeutic application is determined based on the provision of the protein in a dose and volume that is tolerated, and of therapeutic value, for the patient. If a reduced viscosity formulation is to be administered by injection, the protein concentration will depend on the injection volume (usually 1.0 ml-1.2 ml). Protein-based therapies may require several mg / kg of dosages per week, per month, or for several months. Accordingly, if a protein is to be provided at 2-3 mg / kg of the patient's body weight, and average patient weights of 75 kg, 150 mg-225 mg of the protein will be required to be delivered in 1.0 ml-1.2. ml of injection volume. Alternatively, the formulation is provided in a suitable concentration to be distributed at more than one injection site per treatment. When the concentration of the protein in a formulation is increased, the viscosity of the protein formulation is also likely to increase. The increased viscosity of the formulation makes the formulation harder to administer. Accordingly, there is a need to decrease the viscosity of the protein formulations when the increased viscosity impacts its ability to be used. Viscosity Reducing Agents It has been found that the addition of relatively low concentrations of certain viscosity reducing agents to a protein formulation reduces the viscosity of the protein formulation. The term "viscosity reducing agent" as used herein, includes any agent that reduces the viscosity of a protein formulation compared to a protein formulation that does not contain, or that contains a minor amount of, viscosity reducing agent. For example, a viscosity reducing agent generally reduces the viscosity of a protein formulation by about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 90%, or about 95% compared to the viscosity of the protein formulation without, or containing lower amounts of, viscosity reducing agent. For example, a viscosity reducing agent generally reduces the viscosity of the protein formulation by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 90%, or 95%, compared with the viscosity of a protein formulation without, or containing lower amounts of, the viscosity reducing agent. Non-limiting examples of viscosity reducing agents they include calcium chloride, magnesium chloride, arginine hydrochloride, sodium chloride, sodium thiocyanate, ammonium thiocyanate, ammonium sulfate, sodium phosphate, and ammonium chloride. In one embodiment, the viscosity reducing agent is calcium chloride. In another embodiment, the viscosity reducing agent is magnesium chloride. In an alternative embodiment, more than one viscosity reducing agent is added to the protein formulation. A viscosity reducing agent is generally added to a protein formulation at a final concentration of between about 1 mM and about 150 mM, for example, between about 1 mM and about 50 mM, between about 2 mM and about 40 mM, between about 4 mM and approximately 30 mM, between approximately 4 mM and approximately 25 mM, between approximately 5 mM and approximately 20 mM, between approximately 5 mM and approximately 25 mM, between approximately 5 mM and approximately 30 mM, between approximately 5 mM and approximately 40 mM, and between approximately 5 mM and approximately 50 mM. In certain embodiments, the viscosity reducing agent is added to the protein formulation at a final concentration of less than 14 mM, less than 13, mM, less than 12 mM, less than 11 mM, less than 10 mM, less than 9 mM , lower than 8 mM, lower than 7 mM, lower of 6 mM, less than 5 mM, less than 4 mM, less than 3 mM, or less than 2 M. In other embodiments, the viscosity reducing agent is added to the protein formulation at a concentration of between 0.5 mM and 14 mM. mM, between 0.5 mM and 13 mM, between 0.5 mM and 12 mM, between 0.5 and 11 mM, between 0.5 mM and 10 mM, between 0.5 mM and 9 mM, between 0.5 mM and 8 mM, between 0.5 mM and 7 mM, between 0.5 mM and 6 mM, or between 0.5 mM and 5 mM. In one embodiment, the viscosity reducing agent is calcium chloride or a final concentration of between about 5 mM and about 20 mM in the formulation. In another embodiment, the viscosity reducing agent is calcium chloride at a concentration of between 5 mM and about 14 mM in the formulation. In other embodiments, the viscosity reducing agent is magnesium chloride at a final concentration of between about 5 mM and about 20 mM in the formulation. In another embodiment, the viscosity reducing agent is magnesium chloride or a final concentration of between 5 mM and about 14 mM in the formulation. The viscosity of a protein formulation can be measured by any suitable method known in the art including, for example, the use of a capillary viscometer or a conical plate rheometer. PH buffers The term "pH buffer" as used in present, includes those agents that maintain the pH in a solution, for example, a formulation, in a desired range. The pH of a formulation as described herein is generally between about a pH of 5.0 to about 9.0, for example, a pH of about 5.5 to about 6.5, about a pH of 5.5 to about 6.0, about a pH of 6.0 to about 6.5, pH 5.5, pH 6.0, or pH 6.5. In general, a buffer is used that can maintain a solution at a pH of 5.5 to 6.5. Non-limiting examples of pH buffers that can be used in a formulation described herein include, histidine, succinate, gluconate, tris (tromethamol), phosphate, citrate, 2-morpholinene-sulfonic acid (MES), sodium phosphate, sodium, and cacodylate. Histidine is a pH buffer that is typically in the reduced viscosity formulations that are not administered by subcutaneous, intramuscular or peritoneal injection. The concentration of the buffer is between about 5 mM and 30 mM. In one embodiment, the pH buffer of a formulation is histidine at a concentration of about 5 mM and about 20 mM. Excipients In the addition to the protein, the reducing agent of viscosity, and a buffer, a reduced viscosity formulation as described herein may also contain other substances. The substances include, but are not limited to, cryoprotectants, lyoprotectants, surfactants, bulking agents, anti-oxidants, and stabilizing agents. In one embodiment, a reduced viscosity protein formulation described herein includes excipients selected from the group consisting of cryoprotectant, a lyoprotectant, a surfactant, a bulking agent, an anti-oxidant, a stabilizing agent, and combinations thereof . The term "cryoprotectant" as used herein, includes agents that provide stability to the protein in a formulation against stresses induced by freezing, for example, preferably being excluded from the protein surface. Cryoprotectants can also offer protection during primary and secondary drying and long-term storage of the product. Non-limiting examples of cryoprotectants include sugars, such as sucrose, glucose, trehalose, mannitol, mannose, and lactose; polymers, such as dextrin, hydroxyethyl starch and polyethylene glycol; surfactants, such as polysorbates (e.g., PS-20 or PS-80); and amino acids, such as glycine, arginine, leucine, and serine. Generally, a cryoprotectant that exhibits low toxicity in biological systems. A cryoprotectant, if included in the formulation, is generally added at a final concentration of between about 0.1% and about 10% (weight / volume), for example between about 0.5% and about 10%, between about 0.5% and about 5%. %, between about 0.5% and about 2%, between about 1% and about 5%, or between about 5% and about 10%. In one embodiment, the cryoprotectant is sucrose at a concentration of between about 0.5% and about 10% (weight / volume). In one embodiment, a lyoprotectant is added to a formulation described herein. The term "lyoprotectant" as used herein, includes agents that provide stability to the protein during the lyophilization or dehydration process (primary and secondary lyophilization cycles), for example by providing an amorphous vitreous matrix and through binding to the protein through hydrogen bonding, which replaces the water molecules that are removed during the drying process. This helps maintain protein conformation, minimizes protein degradation during the lyophilization cycle, and improves long-term product stability. Non-limiting examples of lyoprotectants include sugars, such as sucrose or trehalose; an amino acid, such as monosodium glutamate, lysine or non-crystalline histidine; a methylamine such as betaine; a lyopropy salt, such as magnesium sulfate; a polyol, such as trihydric or higher sugar alcohols, for example, glycerin, erythritol, glycerol, arabitol, xylitol, sorbitol, and mannitol; propylene glycol; polyethylene glycol; pluronic; and combinations of these. The amount of lyoprotectant additional to the formulation is generally an amount that does not lead to an unacceptable amount of degradation / aggregation of the protein when lyophilized to the protein formulation. When the lyoprotectant is a sugar (such as sucrose or trehalose) and the protein is an antibody, non-limiting examples of lyoprotectant concentrations in a protein formulation of reduced viscosity are from about 10 mM to about 400 mM, about 30 mM at about 300 mM, and from about 50 mM to about 100 mM. In certain embodiments, a surfactant is included in a formulation described herein. The term "surfactant" as used herein, includes agents that reduce the surface tension of a liquid through the adsorption of the aerea-liquid interface. Examples of surfactants include, without limitation, nonionic surfactants, such as polysorbates (for example, polysorbate 80 or polysorbate 20); poloxamers (for example, poloxamer 188); Triton ™ (for example, Triton ™ X-100); sodium dodecyl sulfate (SDS), octal glycoside sodium; lauryl-sulfobetaine; myristyl-sulfobetaine; linoleyl-sulfobetaine; stearyl-sulfobetaine lauryl-sarcosine; myristyl sarcosine; linoleil-sarcosine stearil-sarcosine; linoleil-betaine; myristyl betaine cetyl betaine; lauroamidopropyl betaine; cocamidopropyl betaine; Linoleamidopropyl betaine; Myristamidopropyl betaine, palmidopropyl betaine, isostearamide propyl betaine (for example, lauroamidopropyl); miristarnidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; methyl sodium cocoyl, or disodium methyl ofeil-taurate; and the series Monaquat ™ (Mona Industries, Inc., Paterson, N.J.); polyethylene glycol; polypropylene glycol; and ethylene and propylene glycol copolymers (e.g., pluronics, PF68). The amount of surfactant added is such as to maintain the aggregation of the reconstructed protein at an acceptable level as tested using for example, SEC-HPLC to determine the percentage of high molecular weight species (HMW) or low molecular weight species (LMW), and minimizes particle formation after reconstitution of a lyophilate of a protein formulation described herein. For example, the surfactant may be present in a formulation (liquid or before lyophilization) in an amount of about 0.001-0.5%, for example, of about 0.05-0.3%. In some embodiments, the bulking agent is included in a reduced viscosity formulation. The term "bulk agent" as used herein, includes agents that provide the structure of the lyophilized product without interacting directly with the pharmaceutical product. In addition to providing a pharmaceutically elegant cake, the bulking agents can also impart useful qualities with respect to the modification of the collapse temperature, provide freeze-thaw protection and improve the stability of the protein in long-term storage. Non-limiting examples of bulking agents include mannitol, glycine, lactose and sucrose. The bulking agents may be crystalline (eg, glycine, mannitol, or sodium chloride) or amorphous (such as dextrin or hydroxyethyl starch) and are generally used in protein formulations in an amount of 0.5% to 10%. Other pharmaceutically acceptable carriers, excipients, or stabilizers, such as those described in Remington: The Science and Practice of Pharmacy 20th edition, Gennaro, Ed., Lippincott Williams & Wilkins (2000) can also be included in a protein formulationyou. described herein, as long as they do not adversely affect the desired characteristics of the formulation. Acceptable carriers, excipients, or stabilizers are non-toxic to recipients (e.g., patients) at dosages and concentrations used and include: additional pH buffering agents; conservatives, co-solvents; antioxidants, including ascorbic acid and methionine; chelating agents such as EDTA; metal complexes (e.g., Zn protein complexes); biodegradable polymers, such as polyesters; salt-forming counterions, such as sodium, polyhydric sugar alcohols; amino acids, such as alanine, glycine, glutamine, asparagine, histidine, arginine, lysine, ornithine, leucine, 2-phenylalanine, glutamic acid, and trionine; organic sugars or sugar alcohols, such as lactitol, stachyose, mannose, sorbose, xylose, ribose, ribitol, myoinisitose, myoinisitol, galactose, galactitol, glycerol, cyclitol (e.g., inositol), polyethylene glycol; reducing agents containing sulfur, such as urea, glutathione, thioctic acid, sodium thioglycolate, thioglycerol, α-monothioglycerol, and sodium thiosulfate; low molecular weight proteins, such as human serum albumin, bovine serum albumin, gelatin or other immunoglobulins; and hydrophilic polymers, such as polyvinyl pyrrolidone.

MYO-029 Protein Usage Forms In one example, a reduced viscosity formulation MYO-029 can be formulated using 1 mg / ml to 300 mg / ml antibody MYO-029. The MYO-029 formulation generally includes between about 1 mM and about 50 mM calcium chloride or magnesium chloride. The formulation may include about 5 mM to about 25 mM histidine. The formulation may include about 1% to about 5% (w / v) of sucrose or trehalose. In some cases, the formulation may include from about 10 mM to about 25 mM methionine. In certain MYO-029 formulations, 0.05-0.2% (w / v) of polysorbate-20 or polysorbate-80 is added. The pH of the formulation is generally between 5.5 and 6.5. In a specific example, the formulation MYO-029 comprises 150 mg / ml of the MYO-029 antibody, 10 mM of calcium chloride or magnesium chloride, 20 mM of histidine, 4% of sucrose, and has a pH of 6.0. In another specific example, the formulation MYO-029 comprises 75 mg / ml of MYO-029 antibody, 5 mM of calcium chloride or magnesium chloride, 10 mM of histidine, 10 mM of methionine, 2% of sucrose, and has a pH of 6.0. In another specific example, a MYO-029 antibody formulation comprises 150 mg / ml MYO-029 antibody, 10 mM magnesium chloride or calcium chloride, 20 mM histidine, 20 mM methionine, 4% of sucrose, 0.2% polysorbate-80, and has a pH of 6.0. MYO-028 Reduced viscosity formulations MYO-028 can be formulated using 1 mg / ml to 300 mg / ml antibody MYO-028. Formulation MYO-028 generally includes between about 1 mM and about 50 mM calcium chloride or magnesium chloride. The formulation may include about 5 mM to about 25 mM histidine. The formulation may include about 1% to about 5% (w / v) of sucrose or trehalose. In some cases, the formulation may include from about 10 mM to about 25 mM methionine. The pH of a MYO-028 formulation is generally between about 5.5 and 6.5. In a specific example, a MYO-028 antibody formulation comprises 50 mg / ml antibody, 10 mM histidine, 5% sucrose, and has a pH of 6.5. In another specific example, a MYO-028 antibody formulation comprises 50 mg / ml of the antibody, 10 mM of calcium chloride or magnesium chloride, 10 mM of histidine, 5% of sucrose, and has a pH of 6.5. J695 Reduced viscosity formulations J695 can be formulated using 1 mg / ml to 300 mg / ml J695 antibody. A J695 formulation generally includes between about 1 mM and about 50 mM chloride of calcium or magnesium chloride. The formulation may include about 5 mM to about 25 mM histidine. The formulation may include between about 1% to about 5% (w / v) of sucrose or trehalose. In some cases, the formulation may include about 10 mM to about 25 mM methionine. In certain J695 formulations, between about 1% to about 5% (w / v) of mannitol is added. The pH of the formulation is generally between 5.5 and 6.5. In a specific example, the J695 antibody formulation comprises 100 mg / ml J695 antibody, 10 mM histidine, 10 mM methionine, 4% mannitol, 1% sucrose, and has a pH of 6.0. In another specific example, a J695 antibody formulation comprises 100 mg / ml of J695 antibody, 10 mM histidine, 10 mM methionine, 5 mM methionine, 5 mM calcium chloride or magnesium chloride, 4% mannitol, 1% sucrose, and has a pH of 6.0. In another specific embodiment, the J695 antibody formulation comprises 100 mg / ml J695 antibody, 10 mM histidine, 10 mM methionine, 10 mM calcium chloride or magnesium chloride, 4% mannitol, 1% sucrose, and it has a pH of 6.0. IMA-638 Formulations of the IMA-638 protein can be formulated using 1 mg / ml to 300 mg / ml of the IMA-638 antibody. A reduced viscosity formulation containing IMA- 638 generally includes between about 1 mM and about 50 mM calcium chloride or magnesium chloride. The formulation may include about 5 mM to about 25 mM histidine. The formulation may also include between about 1% to about 10% (w / v) of sucrose or trehalose. The pH of the formulation is generally between 5.5 and 6.5. In a specific example, the IMA-638 antibody formulation comprises 50 mg / ml of the IMA-638 antibody, 10 mM of histidine, 10 mM of methionine, 5% of sucrose, and has a pH of 6.0. In another specific example, the IMA-638 antibody formulation comprises 100 mg / ml of the IMA-638 antibody, 20 mM of histidine, 10% of sucrose, and has a pH of 6.0. In another specific example, the IMA-638 antibody formulation comprises 50 mg / ml of the IMA-638 antibody, 5 mM of calcium chloride or magnesium chloride, 10 mM of histidine, 10% of sucrose, and has a pH of 6.0 . In yet another specific example, the IMA-638 antibody formulation comprises 100 mg / ml of the IMA-638 antibody, 10 mM calcium chloride or magnesium chloride, 20 mM histidine, 10% sucrose, and has a pH of 6.0. Storage Methods A reduced viscosity protein formulation described herein can be stored by any suitable method known to one skilled in the art. Non-limiting examples of methods for preparing a reduced viscosity formulation for storage include freezing, lyophilization, and spray drying the protein formulation. In some cases, a reduced viscosity formulation is frozen for storage. Accordingly, it is desirable that the formulation be relatively stable under the conditions, including when subjected to freeze-thaw cycles. One method for determining the viability of a formulation for frozen storage is to subject a sample formulation to at least 2, for example, three to ten freezing cycles (a, for example -20 ° C or -80 ° C) and defrosting (for example, through rapid thawing at room temperature or slow thawing on ice), determining the amount of LMW species and / or HMW species that accumulate after the freeze-thaw sites and comparing the number of LMW species or HMW species present in the sample before the freeze-thaw procedure. An increase in the LMW species or in the HMW species indicates a decreased stability of the stored protein as part of the formulation. Liquid chromatography or high-throughput size exclusion (SEC-HPLC) can be used to determine the presence of LMW and HMW species. A suitable formulation can accumulate undesirable HMW species or LMW species, but to the extent that the presence of HMW species or LMW species make the formulation inadequate for its intended use. In some cases, a formulation is stored as a liquid. Accordingly, it is desirable that the liquid formulation be relatively stable under the conditions, including at various temperatures. One method for determining the viability of a liquid storage formulation is to store the sample formulation at various temperatures (such as 2-8 ° C, 15 ° C, 20 ° C, 25 ° C, 30 ° C, 35 ° C, 40 ° C, and 50 ° C) and monitor the amount (for example, the change in the percentage) of the HMW species and / or LMW species that accumulate over time. Additionally, the protein loading profile can be monitored through high performance liquid chromatography-cation exchange (CEX-HPLC). In general, the percentage of high molecular weight species or low molecular weight species is determined either as a percentage of the total protein content in a formulation or as a change in the percentage increase over time (ie say, during storage) as appropriate for the assay of the parameter being determined. In general, and in the non-limiting examples, the change in the percentage of the protein in high molecular weight species or low molecular weight species in a reduced viscosity formulation is not greater than 10%, for example, no more than about 8%, no more than about 5%, or no more than about 3% with respect to the parameter tested (eg, time, temperature, additional compounds in the formulation, lyophilization or agitation). Alternatively, a formulation can be stored after lyophilization. The term "lyophilization" as used herein, refers to a process by which the material to be dried is first frozen, followed by the removal of the ice or the frozen solvent by sublimation in a vacuum environment. An excipient (eg, lyoprotectant) can be included in the formulation to be lyophilized to thereby improve the stability of the lyophilized product in storage. The term "reconstituted formulation" as used herein, refers to a formulation that has been prepared through dissolution of a lyophilized protein formulation in a diluent such that the protein is dispersed in the diluent. The term "diluent" as used herein, is a substance that is pharmaceutically acceptable (safe and non-toxic for administration to a human) and is useful for the preparation of a liquid formulation, such as a reconstituted formulation after administration. lyophilization. Non-limiting examples of diluents include sterile water, water bacteriostatic for injection (BWFI), a solution regulated in its pH (for example, saline regulated in its pH with phosphate), sterile saline solution, Ringer's solution, dextrose solution, or aqueous solutions of salts and / or pH buffers. The test of a reduced viscosity formulation for the stability of the protein component of the formulation after lyophilization is useful for determining the viability of a formulation. The method is similar to that described above for freezing, except that the sample formulation is lyophilized instead of frozen, reconstituted using a diluent and the reconstituted formulation is tested for the presence of the LMW species and / or HMW species. An increase in the LMW species or HMW species in the lyophilized sample compared to a corresponding sample formulation that was not lyophilized indicates a decreased stability in the lyophilized sample. In some cases, a formulation was spray-dried and then stored. For spray drying, a liquid formulation is converted into an aerosol in the presence of a dry gas stream. The water is removed from the droplets of the formulation in a gas stream, resulting in dried particles of the drug formulation. The excipients can be included in the formulation to (1) protect the protein during dehydration from spray drying, (2) protect the protein during storage after spray drying, and / or (3) give the solution suitable properties for aerosolization. The method is similar to that described above for freezing, except that the sample formulation is spray-dried instead of frozen, reconstituted in a diluent and the reconstituted formulation is tested for the presence of LMW species and / or HMW species. An increase in the LMW or HMW species in the spray-dried sample compared to a corresponding sample formulation that was not lyophilized indicates a decreased stability in the spray-dried sample. Methods of Treatment The reduced viscosity formulations described herein are useful as pharmaceutical compositions in the treatment and / or prevention of diseases or disorders in a patient in need thereof. The term "treatment" refers to both therapeutic and prophylactic treatment and preventive treatment. The treatment includes the application or administration of the reduced viscosity formulation to the body, an isolated tissue, a cell of a patient having a disorder, a symptom of a disorder, who is at risk of a disorder, or a predisposition toward a disorder , for the purpose of healing, heal, relieve, release, alter, remedy, decrease, improve, or affect the disorder, the symptom of the disorder, or the predisposition toward the disorder. Those "in need of treatment" include those who already have a disorder, as well as those where the disorder is to be avoided. The term "disorder" is any condition that could benefit from treatment with a protein formulation described herein. This includes chronic or acute disorders or diseases including those pathological conditions that predispose the subject (patient) to the disorder in question. Non-limiting examples of disorders to be treated using a formulation described herein include autoimmune disorders, inflammatory disorders, muscle wasting disorders, allergies, cancers, muscular dystrophy, sarcopenia, cachexia, type II diabetes, rheumatoid arthritis, Crohn's disease. , psoriasis, psoriatic arthritis, asthma, dermatitis, allergic rhinitis, chronic obstructive pulmonary disease, eosinophilia, fibrosis, and excess production of mucus. In one embodiment, the reduced viscosity formulation suitable for use as a pharmaceutical composition comprises an anti-myostatin antibody, and a viscosity reducing agent. In one embodiment, the anti-myostatin antibody is MYO-029. In other embodiments, the anti-myostatin antibody is MYO-022 or MYO-028. The anti-antibody Myostatin is generally at a concentration of approximately 0.5 mg / ml and approximately 300 mg / ml in the formulation. In another embodiment, the viscosity reducing agent is at a final concentration of between about 0.5 mM and 20 mM in the pharmaceutical composition. In another embodiment, the viscosity reducing agent is at a final concentration of between about 0.5 mM and 14 mM in the pharmaceutical composition. In another embodiment, the pharmaceutical composition comprises an anti-myostatin antibody, a viscosity reducing agent, and a pH buffer wherein the pH of the formulation is between about 5.5 to about 6.5. The pharmaceutical compositions described herein may also contain other proteins, drugs, and / or excipients. In particular, other proteins or substances useful for treating the disorder in question can be added to the formulation. Pharmaceutical compositions containing the anti-myostatin antibody are useful in the treatment or prevention of disorders such as, but not limited to, muscle wasting disorder, muscular dystrophy, sarcopenia, cachexia, and type II diabetes. In another embodiment, a pharmaceutical composition comprises an anti-IL-12 antibody and a viscosity reducing agent. In one embodiment, the anti-IL-12 antibody is J695. The anti-IL-12 antibody is generally at a concentration of between about 0.5 mg / ml and about 300 mg / ml in the formulation. In another embodiment, the viscosity reducing agent is at a concentration of between about 0.5 mM and 20 mM in the pharmaceutical composition. In another embodiment, the viscosity reducing agent is at a final concentration of between about 0.5 mM and 14 mM in the pharmaceutical composition. In another embodiment, the pharmaceutical composition comprises an anti-IL-12 antibody, a viscosity reducing agent, and a pH buffer, wherein the pH of the formulation is between about 5.5 to about 6.5. The pharmaceutical compositions described herein may also contain other proteins, drugs, and / or excipients. In particular, the other proteins or substances useful for treating the disorder in question can be added to the formulation. Pharmaceutical compositions containing the anti-IL-12 antibody are useful in the treatment or prevention of disorders such as, but not limited to, autoimmune diseases, inflammatory diseases, rheumatoid arthritis, Crohn's disease, psoriasis, and psoriatic arthritis. In another embodiment, a pharmaceutical composition comprises an anti-IL-13 antibody and a viscosity reducing agent. In one embodiment, the anti-IL-13 antibody is IMA-638. The anti-IL-13 antibody is generally at a concentration of between about 0.5 mg / ml and about 300 mg / ml in the formulation. In another embodiment, the viscosity reducing agent is at a final concentration of between about 0.5 mM and 20 mM in the pharmaceutical composition. In another embodiment, the viscosity reducing agent is at a final concentration of between about 0.5 mM and 14 mM in the pharmaceutical composition. In another embodiment, the pharmaceutical composition comprises an anti-IL-13 antibody, a viscosity reducing agent, and a pH buffer, wherein the pH of the formulation is between about 5.5 to about 6.5. The pharmaceutical compositions described herein may also contain other proteins, drugs, and / or excipients. In particular, the other proteins or substances useful for treating the disorder in question can be added to the formulation. Pharmaceutical compositions containing the anti-IL-13 antibody are useful in the treatment or prevention of disorders such as, but not limited to, asthmatic disorders, atopic disorders, chronic obstructive pulmonary disease, conditions involving airway inflammation, eosinophilia , fibrosis, and excess production of mucus, inflammatory conditions, autoimmune conditions, tumors and cancers, and viral infection.

Administration A reduced viscosity formulation described herein may be administered to a subject in need of treatment using methods known in the art, such as through single or multiple boluses or infusions for a long period of time in a suitable manner, for example, injection or infusion through subcutaneous, intravenous, intraperitoneal, intramuscular, intraarterial, intralesional, or intraarticular administration, topical administration, inhalation, or via sustained release or extended release means. If the formulation has been lyophilized, the lyophilized material is first reconstituted in an appropriate liquid before administration. The lyophilized material can be reconstituted in, for example, BWFI, saline regulated at its pH with phosphate, or the same protein formulation that has been prior to lyophilization. Parenteral compositions can be prepared in unit dosage form for easy administration and uniformity of dosage. "Unit dosage form" as used herein, refers to physically discrete units suitable as unitary dosages for the subject to be treated; each unit contains a predetermined amount of an active compound calculated to produce the desired therapeutic effect in association with the selected pharmaceutical carrier. In the case of the inhalation method, such as a metered dose inhaler, the device is designed to distribute an appropriate amount of a formulation. For administration by inhalation, the compounds are delivered in the form of an aerosol spray from a pressurized container or dispenser containing a suitable propellant, for example, a gas, such as carbon dioxide, or a nebulizer. Alternatively, the inhaled dosage form can be provided as a dry powder using a dry powder inhaler. A reduced viscosity formulation can also be entrapped in microcapsules prepared, for example, by preservation techniques or through interfacial polymerization, for example capsules of hydroxymethylcellulose or gelatin and poly (methylmethacrylate) microcapsules, respectively, in distribution systems of colloidal drugs (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in macroemulsions. The techniques are described in Remington's Pharmaceutical Sciences, 20a. edition. { supra). Sustained-release preparations of the protein formulations described herein may also be prepared. Suitable examples for sustained release preparations include semi-permeable matrices of hydrophobic solid polymers containing the protein formulation. Examples of sustained release matrices include polyesters, hydrogels (e.g., poly (2-hydroxyethyl-methacrylate), or poly (vinylalcohol)), polylactides, L-glutamic acid copolymers, and? -ethyl-L-glutamate, non-degradable ethylene vinyl, degradable lactic acid-glycolic acid copolymers, and poly-D- (-) - 3-hydroxybutyric acid. Sustained-release formulations of the proteins described herein can be developed using polylactic-coglycolic acid (PLGA) polymer because of their biocompatibility and wide range of biodegradable properties. The degradation products of PLGA, lactic acid and glycolic acid can be rapidly cleared within the human body. In addition, the degradation of this polymer can be adjusted from months to years depending on its molecular weight and composition. The liposomal compositions can also be used to formulate the proteins or antibodies described herein. Dosing The toxicity and therapeutic efficacy of a formulation can be determined through pharmaceutical methods known in the art using, for example, cell cultures or experimental animals, for example, for the determination of LD50 (50% lethal dose). from population) and ED50 (therapeutically effective dose in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index, and can be expressed as the LD50 / ED50 ratio. The data obtained from cell culture assays and animal studies can be used in the formulation of a dosage range for use in humans. The dosage of the formulations is generally based within a range of circulating concentrations that includes the ED50 with little or no toxicity. The dose may vary within this range depending on the dosage form used and the route of administration used. For any formulation used in the method of the invention, the therapeutically effective dose can be estimated initially from the cell culture assays. A dose can be formulated in animal models to achieve a plasma concentration range in circulation that includes the IC50 (ie, the concentration of the test compound that achieves a maximum average inhibition of symptoms) as determined in cell culture. . The information can be used to more precisely determine the useful doses in humans. The levels in the plasma can be measured, for example, through high performance liquid chromatography. The appropriate dosage of the protein of the The formulation will depend on the type of disorder to be treated, the severity and course of the disorder, whether the agent is administered for preventive or therapeutic purposes, the prior therapy, the patient's clinical history, and the response to the agent, and the discretion of the treating physician. . A formulation is generally provided in such a way that the dosage is between 0.1 mg protein / kg body weight to 100 mg protein / kg body weight. The formulation is administered to the patient only once or during series of treatment. In one embodiment, the formulation of the myostatin antibody (eg, MYO-22, MYO-28, MYO-029) is delivered to the patient in need thereof at a dosage of 1 mg / kg to 10 mg / kg of weight bodily. In another embodiment, a formulation of the IL-12 antibody is administered to a patient in need thereof at a dosage of 1 Sf / kg to 5 mg / kg of body weight. In a further embodiment, a formulation of the IL-13 antibody is administered to a patient in need thereof at a dosage of about 0.5 mg / kg to about 5 mg / kg of the patient's body weight. A formulation to be used in in vivo administration must be sterile. A formulation can be rendered sterile for example, through filtration through sterile filtration membranes, before, or after, the formulation of a liquid or lyophilization and reconstitution. The therapeutic compositions described herein are generally placed in a container having a sterile access port, for example, an intravenous solution bag, or a bottle having a pierceable plug through a hypodermic injection needle. Articles of Manufacture In another embodiment, an article of manufacture is provided containing a formulation described herein and typically provides instructions for its use. The article of manufacture comprises a container suitable for containing the formulation. Suitable containers include, without limitation, bottles, bottles (e.g., double-chamber bottles), syringes (e.g., single or double chamber syringes), test tubes, nebulizers, inhalers (e.g. dry powder inhalers), or deposits. The container can be formed from a variety of materials, such as glass, metal or plastic (eg, polycarbonate, polystyrene, polypropylene), the container maintains the formulation and the label on or associated with, the container indicating the directions for the reconstitution and / or use. The label may further indicate that the formulation is useful or intended for subcutaneous administration. The container controlling the formulation can be a multi-purpose bottle, which allows repeated administrations (eg, 2-6 doses) of the formulation. The article of manufacture may further comprise a second container comprising a suitable diluent (eg, WFI, 0.9% NaCl, BWFI, or salt buffered in its pH with phosphate). When the article of manufacture comprises a lyophilized version of a protein formulation, mixing a diluent with the lyophilized formulation will provide a final protein concentration in the reconstituted formulation of generally at least 20 mg / ml. The article of manufacture may also include other desirable materials from a commercial and user standpoint, including other pH buffers, diluents, filters, needles, syringes, package inserts with instructions for use. The invention is further illustrated by the following examples. The examples are provided for illustrative purposes only. They are not constructed as limiting the scope or content of the invention in any way. EXAMPLES Example 1 Viscosity of Antibody Formulations Antibodies of the peptide anti-β-amyloid (anti-AB), anti-IL-13, anti-IL-12 (J695) and anti-myostatin (MYO-029) were formulated as described in Table 1. The The viscosity of these antibody formulations was measured using an Anton Paar Physica MCR301 cone and plate rheometer. Specifically, a CP25-1 cone (24,971 mm in diameter, 1,002 ° angle) was used for all measurements; the constant effort velocity was constant at 898 l / s with a duration of 100 seconds. The measurements were made every 10 seconds. The viscosity measurements were carried out at both 4 ° C and 25 ° C using an automatic Peltier temperature control unit. The liquid sample loaded on the plate was 90 μl. Each sample was analyzed in triplicate. Table 1 below lists the viscosities of different antibodies at different concentrations in different formulations.

The data shown in Table 1 show that the viscosity of anti-myostatin (MYO-029) is significantly higher compared to the other antibodies listed in the Table. The viscosities for all other antibodies were increased to 4 ° C. This increase is proportionally much higher for MYO-029. Example 2 Effect of Various Salts on the Viscosity of an MYO-029 Antibody Formulation An MYO-029 antibody formulation was formulated, at a concentration of 10 mM histidine, 2% sucrose, pH 6.0. Concentrated solutions of salts (e.g., calcium chloride, magnesium chloride, sodium chloride, and sodium biphosphate) were diluted in the MYO-29 antibody formulation using a pipette. The effect of these salts on the viscosity of the MYO-29 antibody formulation was measured as described in Example 1. These data are shown in Fig. 1. Both MgCl 2 and CaCl 2 at concentrations on the scale of about 5 mM at about 20 mM significantly reduced the viscosity of the MYO-29 antibody formulation. NaCl and NaH2P04 on the other hand, had very little effect on this scale. In this way the calcium chloride and magnesium chloride, at concentrations of about 5 mM to about 20 mM, are effective viscosity reducing agents for the formulations of MYO-29 antibody, unlike sodium chloride and sodium bisphosphate. Example 3 Effect of Calcium Chloride in a J695 Antibody Formulation The J695 antibody formulation was measured at two different concentrations of the J695 antibody, ie, 100 mg / ml and 300 mg / ml. The viscosity of the J695 antibody formulation at a higher concentration will be higher than the viscosity of the J695 antibody formulation at a lower concentration. Calcium chloride at a final concentration of about 5 mM to 20 mM was added to the J695 300 mg / ml J695 antibody formulation. In this case the viscosity of the antibody formulation was expected to decrease as compared to the J695 formulation without calcium chloride. Accordingly, calcium chloride, at concentrations of about 5 mM to about 20 mM, is effective as a viscosity reducing agent for J695 antibody formulations. EXAMPLE 4 Effect of calcium chloride on the viscosity of an MYO-028 Antibody Formulation Another MYO-anti-myostatin antibody was concentrated 028 using Centricon Ultrafree®-4 at a concentration of 95 mg / ml. Calcium chloride was added to MYO-028 according to Table 2 below: Table 2 MYO-028 was formulated at 95 mg / ml in 10 mM histidine, 5% sucrose, pH 6.5. The CaCl2 solution consisted of 10 mM histidine, 2% sucrose, 2M CaCl2. The buffer solution consisted of 10 mM histidine, 5% sucrose, pH 6.5. The viscosity of these MYO-028 antibody formulations were measured using the same rheometer method as described in Example 1 with the additional use of a solvent trap to prevent evaporation, a liquid sample load of 100 μl of MYO-028 on the plate, and the test was performed at room temperature. The data from these experiments are shown in Fig. 2. The addition of CaCl2 decreased the viscosity of the antibody formulation MYO-028 at 25 mM and 50 mM CaCl2 compared to a formulation of MYO-028 without CaCl2. These data demonstrate the viability of CaCl2 for use as an agent to reduce the viscosity of a formulation of protein, for example to formulate an antibody formulation of reduced viscosity. Example 5 Effect of Calcium Chloride on the Viscosity of an IMA-638 Antibody Formulation. To test the effect of calcium chloride on the viscosity of an IMA-638 antibody formulation, different amounts of calcium chloride were pipetted into aliquots of the IL-13 antibody, IMA-638. The aliquots of the IMA-638 antibody had a protein concentration of approximately 150 mg / ml. Fig. 3 provides a graphic description of the effect of calcium chloride on the viscosity of the IMA-638 protein formulations. The viscosity of IMA-638 did not show the same reduction in viscosity as was observed for MYO-029.

These data demonstrate a method for identifying a viscosity reducing agent suitable for use with a protein formulation. EXAMPLE 6 Effect of Calcium Chloride on the Stability of the MYO-029 Antibody The addition of a compound (ie, a viscosity reducing agent, for example CaCl 2) to a protein formulation could potentially affect the stability of the molecule towards the stresses induced by freezing / thawing. This effect could be either harmful, beneficial, or have no effect on the stability of the protein during freezing / thawing. To evaluate the effect of an agent (ie, CaCl2) in the freeze / thaw-induced degradation of antibody MYO-029, the molecule was subjected to 10 thawing cycles at -80 ° C and 37 ° C, in the presence or absence of 5 mM CaCl2. Drug substance MYO-029 was formulated in 10 mM histidine, 2% sucrose, in the presence or absence of calcium chloride through ultrafiltration and diafiltration. The concentration of the final protein was approximately 75 mg / ml. Twenty microliters of aliquots were frozen at -80 ° C and thawed at room temperature. This was repeated for 5 and 10 freeze / thaw cycles. The samples were diluted 25 times with pH buffer of formulation and analyzed by measuring the absorbance at 280 nm for the concentration of the protein and SEC-HPLC for the percentage of products with high molecular weight (% of HMW). The effect of the degradation induced by freezing / thawing was evaluated by (i) protein recovery (absorbance at 280 nm), and (ii) percentage of high molecular weight formation (% of HMW) determined by high performance liquid chromatography- size exclusion (SEC-HPLC). The HMW formation is the most common degradation path for these molecules. The results of these studies are shown in Fig. 4A and Fig. 4B. Compared with the corresponding control sample without CaCl2, the addition of 5 mM CaCl2 to the formulation had no effect on protein recovery or% HMW formation. In this way, the addition of calcium chloride seems to have no impact on the stability of the MYO-29 antibody formulation. This indicates that the stability of CaCl 2 for use as a viscosity reducing agent in a protein formulation, for example, in a reduced viscosity antibody formulation.

Example 7 Effect of calcium chloride on the stability of an MYO-29 antibody formulation Addition of CaCl2 to a protein formulation could potentially affect the liquid stability of the molecules over time. This effect could be either harmful, beneficial, or having no effect on the stability of the protein during storage. To evaluate the effect of this agent on the liquid stability of MYO-029 in heat-induced degradation, formulations containing MYO-029 were subjected to to storage at 50 ° C for up to seven days. The aliquots were harvested at several points in time and analyzed for protein concentration by absorbance at 280 nm and% HMW was analyzed by SEC-HPLC. The data is shown in Fig. 5A and Fig. 5B. Compared with the control sample, the addition of CaCl2 in the formulation did not have a negative effect on the stability of the protein in the liquid state stored at 50 ° C. The percentage of HMW in the drug substance also appears to be slightly lower in the material containing CaCl2. These data also demonstrate the feasibility of using CaCl2 as a viscosity reducing agent. They also demonstrate a method for determining the viability of an agent that reduces the viscosity of a protein formulation, for example, with respect to whether the agent has an effect on the stability of the protein formulation. Example 8 Effect of Calcium Chloride on Stability of Lyophilized MYO-029 The addition of an agent such as CaCl2 to a protein formulation could potentially affect the lyophilized dosage forms of the protein over time. This effect could be either harmful, beneficial or having no effect on the stability of the protein during storage. To evaluate the effect of this excipient in the stability of lyophilized MYO-029, a formulation containing the molecule, was freeze-dried both with and without 5 mM CaCl2 (control) and subjected to storage at 50 ° C and 4 ° C for four weeks. The flasks were extracted weekly and analyzed for protein concentration by absorbance at 280 nm, percentage of HMW by SEC-HPLC, and charge distribution by high performance liquid chromatography-cation exchange (CEX-HPLC). The volume extracted and the viscosity (a point in time only) were also measured. A. Viscosity of Reconstituted Drug Product The viscosity of drug product MYO-029 (which was measured substantially in the same manner as in Example 1) at about 150 mg / ml was reduced when 5 mM calcium chloride was present. in the formulation. B. Volume Removed from the Bottle The amount of drug product that can be removed from the vial with a 1 ml syringe and 21 G needle was improved when CaCl2 is present in the formulation. C. Protein Concentration Compared to the control, the addition of CaCl2 to the formulation does not affect the recovery of the protein. D. At Molecular Weight Five mM of CaCl2 will not have any significant effect on the percentage of HMW species that form after four weeks in storage at 4 ° C. However, at 50 ° C, the degree of HMW formation is expected to be significantly reduced compared to the control. E. Load Distribution The points in the stability time were analyzed by CEX-HPLC, a chromatographic tool used to study charge differences in proteins. In In CEX-HPLC, the most negatively charged molecules elute before the more positively charged molecules. This method is used to detect the deamidation of asparagine residues for either aspartic or iso-aspartic acid. Deamidation results in an increase in the net negative charge of the protein, and will elute earlier from the HPLC column. In this experiment, the effect of calcium chloride on the degradation of the protein was investigated resulting in a load change different from the control. Compared to the control without calcium chloride, the same load changes are expected to occur over time. In this way, CaCl2 is expected to have no effect on the load distribution of MYO-029 at both storage temperatures. In summary, compared to the control sample, the addition of CaCl2 to a formulation will not have a significant negative effect on the stability of the protein in the formulation in relation to the experimental control without calcium chloride in the lyophilized state when stored at 4 ° C and 50 ° C. In some cases, it was found that CaCl2 is beneficial for the stability of the protein. OTHER MODALITIES It is understood that since the invention has been described together with the detailed description thereof, the above description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (41)

2. Having described the invention as above, the content of the following claims is claimed as property: 1. A method for reducing the viscosity of a protein formulation, characterized in that it comprises: (a) providing a protein formulation; and (b) adding calcium chloride or magnesium chloride to the protein formulation at a concentration of about 0.5 mM and about 20 mM, wherein the viscosity of the protein formulation with calcium chloride or magnesium chloride is reduced compared with the viscosity of the protein formulation without the viscosity reducing agent. 2. The method according to claim 1, characterized in that the protein is selected from the group consisting of an antibody, an Ig fusion protein, a receptor, a transcription factor, an enzyme, a ligand, and a biologically fragment. active thereof.
3. 3. The method according to claim 1, characterized in that the protein is an antibody or a biologically active fragment thereof.
4. 4. - The method according to claim 1, characterized in that the protein is a fusion protein ig-
5. 5. The method according to claim 1, characterized in that the protein is an antibody and the antibody is an anti-myostatin antibody, an anti-IL-12 antibody, or an anti-IL-13 antibody.
6. 6. - The method according to claim 5, characterized in that the anti-myostatin antibody is MYO-029, wherein the anti-IL-12 antibody is J695, and wherein the anti-IL-13 antibody is IMA-638 .
7. 7. - The method of compliance with the claim 1, characterized in that the viscosity of the protein formulation is reduced by at least about 5% compared to the viscosity of the formulation in the absence of the viscosity reducing agent.
8. 8. A reduced viscosity formulation, characterized in that it comprises: (i) a protein; (ii) a viscosity reducing agent at a concentration between about 5MM and about 20mM in the formulation, wherein the viscosity reducing agent is not sodium chloride or sodium bisphosphate; and (iii) a pH buffer; wherein the pH of the formulation is about 5.5-6.5.
9. 9. - The reduced viscosity formulation of according to claim 8, characterized in that the protein is selected from the group consisting of an antibody, an Ig fusion protein, a receptor, a transcription factor, an enzyme, a ligand, and biologically active fragments thereof.
10. 10. The method according to claim 8, characterized in that the protein is an antibody or a biologically active fragment thereof.
11. 11. The method according to claim 8, characterized in that the protein is an Ig fusion protein.
12. 12. - The protein formulation according to claim 8, characterized in that the viscosity reducing agent is calcium chloride, or magnesium chloride.
13. 13. An anti-myostatin antibody formulation, characterized in that it comprises: (i) an anti-myostatin antibody, or a myostatin binding fragment thereof; (ii) a viscosity reducing agent; and (iii) a pH buffer, wherein the pH of the formulation is about 5.5-6.5.
14. 14. - The method according to claim 13, characterized in that the anti-myostatin antibody is a monoclonal antibody.
15. 15. - The method according to claim 14, characterized in that the anti-myostatin antibody is a humanized monoclonal antibody.
16. 16. The method according to claim 14, characterized in that the anti-myostatin antibody binds to myostatin with a Kd of approximately 6 X 10"11 M determined by Biacore ™
17. 17. - The method according to the claim 14, characterized in that the anti-myostatin antibody is selected from the group consisting of MYO-022, MYO- 028, and MYO-029.
18. 18. - The method according to claim 13, characterized in that the anti-myostatin in the formulation is at a concentration of about 25 mg / ml to about 400 mg / ml.
19. 19. The method according to claim 13, characterized in that the viscosity reducing agent is calcium chloride or magnesium chloride.
20. 20. The method according to claim 13, characterized in that the viscosity reducing agent is calcium chloride at a concentration of about 5 mM to about 20 mM.
21. 21. The method according to the claim 13, characterized in that the pH buffer is histidine buffer at a concentration in the formulation from about 4 mM to about 60 mM.
22. 22. The method according to claim 13, characterized in that the formulation also comprises a cryoprotectant.
23. 23. - The method according to the claim 22, characterized in that the cryoprotectant is sucrose or trehalose at a concentration in the formulation of about 0.5% to about 5% (weight / volume).
24. 24. - The method according to claim 13, characterized in that the formulation further comprises a surfactant at a concentration in the formulation of about 0% to 0.2% (weight / volume).
25. 25. The method according to claim 24, characterized in that the surfactant is polysorbate-20 or polysorbate-80.
26. 26. The method according to claim 13, characterized in that the formulation also comprises an anti-oxidant.
27. 27. The method according to claim 26, characterized in that the anti-oxidant is methionine, and the concentration of methionine in the formulation is between approximately 2 mM and approximately 20 mM.
28. 28. The formulation of the anti-myostatin antibody according to claim 13, characterized in that: (i) the anti-myostatin antibody is a fully humanized anti-myostatin antibody in the concentration of about 20 mg / ml to about 400 mg / ml; (ii) the viscosity reducing agent is calcium chloride or magnesium chloride at a concentration of about 20 mg / ml to about 400 mg / ml; and (iii) the buffer is histidine buffer at a concentration of about 5 mM to about 20 mM; wherein the pH of the formulation is about 6.0.
29. 29. The formulation according to claim 13, characterized in that it is lyophilized.
30. 30. The formulation according to claim 13, characterized in that the viscosity of the formulation is reduced by at least about 5% compared to a formulation lacking viscosity reducing agent.
31. 31.- A pharmaceutical composition for the treatment of a disorder selected from the group consisting of muscular dystrophy, sarcopenia, cachexia, and type II diabetes, characterized in that it comprises an anti-myostatin antibody formulation of claim 9.
32. 32. - A method for treating a disorder selected from the group consisting of muscular dystrophy, sarcopenia, cachexia, and type II diabetes, the method characterized in that it comprises administering a therapeutically effective amount of an antibody formulation comprising: (i) an antibody of anti-myostatin or a miostatin binding fragment thereof; (ii) a viscosity reducing agent; and (iii) a pH buffer, wherein the pH of the formulation is about 5.5-6.5.
33. 33. - The method of compliance with the claim 32, characterized in that the antibody formulation is administered by injection, intravenous infusion, or pulmonary administration through a nebulizer or as a dry powder.
34. 34. - A kit characterized in that it comprises a container containing the formulation according to claim 1.
35. 35.- The kit according to claim 34, characterized in that it also comprises directions for the administration of the formulation.
36. 36.- A method to reduce the viscosity of a protein formulation, the method characterized in that comprises: (i) providing a protein formulation; and (ii) adding a viscosity reducing agent to the protein formulation, wherein the viscosity reducing agent is calcium chloride or magnesium chloride, and wherein the concentration of the viscosity reducing agent in the formulation is between about 1. mM and approximately 25 mM.
37. 37.- A method for identifying a protein formulation of reduced viscosity, the method characterized in that it comprises: (i) providing a protein formulation; (ii) adding a viscosity reducing agent to the protein formulation, wherein the viscosity reducing agent is calcium chloride or magnesium chloride, and wherein the concentration of the viscosity reducing agent in the formulation is between about 1 mM and about 25 mM, therefore forming a formulation of reduced potential viscosity; and (iii) determining the viscosity of the potential reduced viscosity protein formulation, wherein when the viscosity of the protein formulation of reduced potential viscosity is reduced compared to the viscosity of the protein formulation without the viscosity reducing agent, the formulation is a formulation of reduced viscosity.
38. 38.- The method according to claim 1, characterized in that the method further comprises determining the stability of the protein formulation.
39. 39.- The method according to the claim 27, characterized in that the method further comprises determining the stability of the protein formulation.
40. 40.- The method according to claim 39, characterized in that the stability of the protein formulation is determined after the freeze-thaw of the protein formulation, after storage of the protein formulation at 50 ° C for 1- 7 days, or after lyophilization.
41. 41.- The method according to claim 39, characterized in that the stability is determined by testing the percentage of high molecular weight species, and percentage of low molecular weight species, or the distribution of the charge of the protein formulation compared to a control .