Patent Description:
The presented subject matter relates to the field of pharmaceutical compositions of antigen binding polypeptides and methods of reducing viscosity of such compositions. Specifically, disclosed herein are excipients that reduce viscosity of pharmaceutical compositions. Furthermore, the disclosed subject matter presents methods related to making such pharmaceutical compositions.

Certain therapeutic polypeptides can be difficult to formulate such that an optimal viscosity is attained, whether because of the nature of the therapeutic polypeptide itself, or because of the concentration (high) of the therapeutic polypeptide, or even both. High viscosity formulations are difficult to handle during formulation and packaging. Furthermore, such preparations can be difficult to administer optimally to a patient, and such administration can be uncomfortable for the patient. The need to identify compounds that are useful for reducing viscosity of highly concentrated protein formulations, to develop methods of reducing the viscosity of such formulations, and to provide pharmaceutical formulations with reduced viscosity are well known in the pharmaceutical arts.

N-acetyl arginine, N-acetyl lysine, N-acetyl histidine, N-acetyl proline, and mixtures thereof, for reducing the viscosity of protein formulation, and methods thereof, are disclosed in <CIT>. The use of arginine dipeptides for reducing the viscosity of liquid formulations is disclosed in <CIT>, <CIT> and <CIT>, and <CIT> discloses the use of L-arginine to reduce aggregation of polypeptides in liquid formulations. N-acetyl-proline-arginine dipeptide was disclosed in <NPL> as a part of a library of peptides used as inhibitors of angiotensin-converting enzyme. However, none of these documents discloses N-acetyl-serine-arginine, N-acetyl-proline-arginine, or N-acetyl-proline-arginine-NH<NUM> dipeptides for reducing viscosity of therapeutic polypeptide formulations.

In a first aspect, provided herein are liquid pharmaceutical compositions comprising a therapeutic polypeptide, such as an antibody or an antigen-binding fragment thereof; a buffer, and at least one N-acetyl-dipeptide, wherein the N-acetyl-dipeptide is N-acetyl-serine-arginine, N-acetyl-proline-arginine, or N-acetyl-proline-arginine-NH<NUM>. In one sub-aspect, the pH of the composition is <NUM> to <NUM>. The therapeutic polypeptide, such as an antibody or antigen-binding fragment thereof, can be present in a concentration of at least <NUM>/ml, at least <NUM>/ml, at least <NUM>/ml, at least <NUM>/ml, at least <NUM>/ml, at least <NUM>/ml, at least <NUM>/ml, and at least <NUM>/ml. In the case wherein the therapeutic polypeptide is an antibody, the antibody can be selected from the group consisting of adalimumab, bevacizumab, blinatumomab, cetuximab, conatumumab, denosumab, eculizumab, erenumab, evolocumab, infliximab, natalizumab, panitumumab, rilotumumab, rituximab, romosozumab, and trastuzumab, or an antigen binding fragment thereof, or is an antibody selected from those presented in Table <NUM>, or an antigen binding fragment thereof. In some sub-aspects, the antibody is evolocumab. The N-acetyl-dipeptide can have a concentration from <NUM> to <NUM>, such as from <NUM> to <NUM>, <NUM>, <NUM>, and <NUM>. The N-acetylated dipeptide can be N-acetyl-serine-arginine. The N-acetylated dipeptide can be N-acetyl-proline-arginine. The N-acetylated dipeptide can be N-acetyl-proline-arginine-NH<NUM>. The buffer can be selected from the group consisting of acetate, glutamate, histidine, and phosphate buffers, or a combination thereof, such as acetate. The buffer can be present at a concentration of <NUM> to <NUM>, such as <NUM>. The pH of the compositions can have a pH of <NUM> to <NUM>, such as a pH of <NUM> to <NUM>, and such as a pH of <NUM>. The compositions can further comprise a surfactant, such as a surfactant selected from the group consisting of polyoxyethylenesorbitan monooleate (polysorbate <NUM> or polysorbate <NUM>), polyoxyethylene-polyoxypropylene block copolymer (Poloxamers such as Pluronic® F-<NUM> and other Pluronics®), Sorbitan alkyl esters (Spans®) Polyethylene glycol octylphenyl ethers (Triton X-<NUM>), Polyethylene glycol alkyl ethers (Brij), Polypropylene glycol alkyl ethers, Glucoside alkyl ethers, and D-α-tocopherol polyethylene glycol succinate (vitamin E TPGS). In some aspects, the surfactant is polysorbate <NUM>, such as at a concentration of about <NUM>% (w/v) polysorbate <NUM> or <NUM>% polysorbate <NUM>. The compositions can further comprise a second oligopeptide comprising arginine and consisting of two to ten amino acid residues. The compositions can further comprise an amino acid, N-acetyl-arginine, N-acetyl-lysine, N-acetyl-histidine, N-acetyl-proline or mixtures of any thereof. The amino acid can be arginine or proline.

In a second aspect, provided herein are methods of reducing viscosity in a pharmaceutical composition comprising a therapeutic polypeptide, such as an antibody or an antigen-binding fragment thereof, wherein the method comprises:.

The therapeutic polypeptide, such as an antibody or antigen-binding fragment thereof, is present in a concentration of at least <NUM>/ml, at least <NUM>/ml, at least <NUM>/ml, at least <NUM>/ml, or at least <NUM>/ml. In cases where the therapeutic polypeptide is an antibody, the antibody can be selected from the group consisting of adalimumab, bevacizumab, blinatumomab, cetuximab, conatumumab, denosumab, eculizumab, erenumab, evolocumab, infliximab, natalizumab, panitumumab, rilotumumab, rituximab, romosozumab, and trastuzumab, or an antigen binding fragment thereof, or is an antibody selected from those presented in Table <NUM>, or an antigen binding fragment thereof. In some sub-aspects, the antibody is evolocumab. The N-acetyl-dipeptide can have a concentration from <NUM> to <NUM>, such as from <NUM> to <NUM>, <NUM>, <NUM>, and <NUM>. The N-acetyl-dipeptide can be N-acetyl-serine-arginine. The N-acetyl-dipeptide can be N-acetyl-proline-arginine. The N-acetyl-dipeptide can be N-acetyl-proline-arginine-NH<NUM>. The N-acetyl-dipeptide can be a lyophilized powder prior to being placed in solution. The buffer can be selected from the group consisting of acetate, glutamate, histidine, and phosphate buffers, or a combination thereof, such as acetate. The buffer can be present at a concentration of <NUM> to <NUM>, such as <NUM>. The pH of the compositions can have a pH of <NUM> to <NUM>, such as a pH of <NUM> to <NUM>, and such as a pH of <NUM>. The compositions can further comprise a surfactant, such as a surfactant selected from the group consisting of polyoxyethylenesorbitan monooleate (polysorbate <NUM> or polysorbate <NUM>), polyoxyethylene-polyoxypropylene block copolymer (Poloxamers such as Pluronic® F-<NUM> and other Pluronics®), Sorbitan alkyl esters (Spans®) Polyethylene glycol octylphenyl ethers (Triton X-<NUM>), Polyethylene glycol alkyl ethers (Brij), Polypropylene glycol alkyl ethers, Glucoside alkyl ethers, and D-α-tocopherol polyethylene glycol succinate (vitamin E TPGS), such as polysorbate <NUM> or polysorbate <NUM>. In some aspects, the surfactant is polysorbate <NUM>, such as at a concentration of about <NUM>% (w/v) polysorbate <NUM> or <NUM>% (w/v) polysorbate <NUM>. The compositions can comprise a second oligopeptide comprising arginine and consisting of two to ten amono acid residues. The compositions can further comprise an amino acid, N-acetyl-arginine, N-acetyl-lysine, N-acetyl-histidine, N-acetyl-proline or mixtures of any thereof. The amino acid can be arginine or proline. The viscosity of the composition may be reduced by at least about <NUM>% or by at least about <NUM>% when compared to a control solution lacking the N-acetyl-dipeptide.

N-acetyl-serine-arginine, N-acetyl-proline-arginine, N-acetyl-proline-arginine-NH<NUM>, and glutamate-arginine dipeptides were found to reduce the viscosity of therapeutic polypeptide formulations, such as those containing antibodies. Surprisingly, while the N-acetyl-dipeptides reduced viscosity to a similar extent as N-acetyl-arginine (NAR), they are significantly more soluble than NAR. Because of their increased solubility, these N-acetyl dipeptides are able to reduce the viscosity of therapeutic proteins even more than NAR because they can be formulated at much higher concentrations.

Both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive. The use of the singular includes the plural unless specifically stated otherwise. The use of "or" means "and/or" unless stated otherwise. The use of the term "including", as well as other forms, such as "includes" and "included", is not limiting. Terms such as "element" or "component" encompass both elements and components comprising one unit and elements and components that comprise more than one subunit unless specifically stated otherwise. The use of the term "portion" can include part of a moiety or the entire moiety. When a numerical range is mentioned, e.g., <NUM>-<NUM>, all intervening values are explicitly included, such as <NUM>, <NUM>, <NUM>, <NUM>, and <NUM>, as well as fractions thereof, such as <NUM>, <NUM>, <NUM>, and <NUM>.

"About" or "~" mean, when modifying a quantity (e.g., "about" <NUM>), that variation around the modified quantity can occur. These variations can occur by a variety of means, such as typical measuring and handling procedures, inadvertent errors, ingredient purity, and the like.

"Comprising" and "comprises" are intended to mean that the formulations and methods include the listed elements but do not exclude other unlisted elements. The terms "consisting essentially of" and "consists essentially of," when used to define formulations and methods include the listed elements, exclude unlisted elements that alter the basic nature of the formulation and/or method, but do not exclude other unlisted elements. So a formulation consisting essentially of elements defined herein would not exclude trace amounts of other elements, such as contaminants from any isolation and purification methods or pharmaceutically acceptable carriers (e.g., phosphate buffered saline), preservatives, and the like, but would exclude, for example, additional unspecified amino acids. The terms "consisting of" and "consists of" when used to define formulations and methods exclude more than trace elements of other ingredients and substantial method steps for administering the compositions described herein. Embodiments defined by each of these transition terms are within the scope of this disclosure.

N-acetyl-Serine and N-acetyl-proline are modified versions of a naturally-occurring amino acids Serine and proline, respectively. N-acetyl-serine and N-acetyl-proline include both D and L forms of the amino acids, such as N-acetyl-L-serine, N-acetyl-D-serine, N-acetyl-L-proline, N-acetyl-D-proline. These N-acetyl-amino acids can be made part of arginine-containing dipeptides. The structure of N-acetyl-serine-arginine dipeptide is shown as structure <NUM>; the structure of N-acetyl-proline-arginine dipeptide is shown as formula <NUM>; the structure of N-actyl-proline-arginine-NH2 is shown as formula <NUM>. Also shown is a glutamate-arginine dipeptide as formula <NUM>. <CHM>
<CHM>
<CHM>
<CHM>.

A "pharmaceutical composition" or a "pharmaceutical formulation" is a sterile composition of (i) a pharmaceutically active drug, such as a biologically active polypeptide, that is suitable for parenteral administration (including intravenous, intramuscular, subcutaneous, aerosolized, intrapulmonary, intranasal and intrathecal administration) to a patient in need thereof and (ii) one or more pharmaceutically acceptable excipients, diluents, and other additives deemed safe by the Federal Drug Administration or other foreign national authorities. Pharmaceutical formulations include liquid (e.g., aqueous) solutions that can be directly administered, and lyophilized powders that can be reconstituted into solutions by adding a diluent before administration. The term "pharmaceutical formulation" specifically excludes, however, compositions for topical administration to patients, compositions for oral ingestion, and compositions for parenteral feeding.

"Viscosity" means a fluid's resistance to flow and can be measured in units of centipoise (cP) or milliPascal-second (mPa-s), where <NUM> cP=<NUM> mPa·s, at a given shear rate. Therefore, hereinafter, a referrence to a viscosity measurement value in cP is to be understood as the time value in mPa·s. Viscosity can be measured by using a rotational viscometer, such as a Brookfield Engineering Dial Reading Viscometer, model LVT, such as a Gemini <NUM> Rheometer (Malvern Instruments) or an AR-G2 Rheometer (TA Instruments). Viscosity can be measured using any other methods and in any other units known in the art (e.g., absolute, kinematic or dynamic viscosity. Regardless of the method used to determine viscosity, the percent reduction in viscosity in excipient formulations versus control formulations remain approximately the same at a given shear rate.

A formulation containing an amount of an excipient effective to "reduce viscosity" (or a "viscosity-reducing" amount or concentration of such excipient) means that the viscosity of the formulation in its final form for administration is at least <NUM>% less than the viscosity of an appropriate control formulation, such as water, buffer, other known viscosity-reducing agents such as salt and the like. Excipient-free control formulations might also be used even if they cannot be implementable as a therapeutic formulation, for example due to hypotonicity.

Likewise, a "reduced viscosity" formulation is a formulation that exhibits lower viscosity compared to a control formulation.

"Stable" formulations of biologically active polypeptides are formulations that exhibit either (i) reduced aggregation and/or reduced loss of biological activity of at least <NUM>% upon storage at <NUM>-<NUM> for at least two years compared with a control formula sample, or (ii) reduced aggregation and/or reduced loss of biological activity under conditions of thermal stress (e.g. <NUM> for one week to <NUM> weeks; <NUM> for one to <NUM> weeks; <NUM> for seven-eight days, etc.). A formulation is considered stable when the polypeptide in the formulation retains physical stability, chemical stability and/or a biological activity.

A polypeptide can be said to "retain its physical stability" in a formulation if, for example, it shows no signs of aggregation, precipitation and/or denaturation upon visual examination of color and/or clarity, or as measured by UV light scattering or by size exclusion chromatography (SEC) or electrophoresis, such as with reference to turbidity or aggregate formation.

A polypeptide can be said to "retain its chemical stability" in a formulation if, for example, the chemical stability at a given time is such that no new chemical entity results from modification of the polypeptide by bond formation or cleavage. Chemical stability can be assessed by detecting and quantifying chemically-altered forms of the polypeptide. Chemical alteration can involve, for example, size modification (e.g., clipping), which can be evaluated using size exclusion chromatography, SDS-PAGE and/or matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI/TOF MS). Other types of chemical alteration include, for example, charge alteration (e.g., resulting from deamidation), which can be evaluated by ion-exchange chromatography. Oxidation is another commonly observed chemical modification.

A polypeptide can be said to "retain its biological activity" in a pharmaceutical formulation relative to unmodified polypeptide if, for example, the percentage of biological activity of the formulated polypeptide (e.g., an antibody) as determined by an assay (e.g., an antigen binding assay) compared to the control polypeptide is between either about <NUM>% to about <NUM>%, about <NUM>% to about <NUM>%, about <NUM>% to about <NUM>%, about <NUM>% to about <NUM>%, or about <NUM>% to about <NUM>%. In some cases, a polypeptide can be said to "retain its biological activity" in a pharmaceutical formulation, if, for example, the biological activity of the polypeptide at a given time is at least <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>% or <NUM>%.

"Antibodies" (Abs) and the synonym "immunoglobulins" (Igs) are glycopolypeptides having the same structural characteristics. While antibodies exhibit binding specificity to a specific antigen, immunoglobulins include both antibodies and other antibody-like molecules that lack antigen specificity. Polypeptides of the latter kind are, for example, produced at low levels by the lymph system and at increased levels by myelomas. Thus, the term "antibody" or "antibody peptide(s)" refers to an intact antibody, an antibody derivative, an antibody analog, a genetically altered antibody, an antibody having a detectable label, an antibody that competes for specific binding with a specified antibody, or an antigen-binding fragment (e.g., Fab, Fab', F(ab')<NUM>, Fv, single domain antibody) thereof that competes with the intact antibody for specific binding and includes chimeric, humanized, fully human, and bispecific antibodies. In some cases, antigen-binding fragments are produced, for example, by recombinant DNA techniques. In other cases, antigen-binding fragments are produced by enzymatic or chemical cleavage of intact antibodies. Antigen-binding fragments include Fab, Fab', F(ab)<NUM>, F(ab')<NUM>, Fv, and single-chain antibodies.

Monoclonal antibodies and antibody constructs include "chimeric" antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is/are identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity. Chimeric antibodies include "primitized" antibodies comprising variable domain antigen-binding sequences derived from a non-human primate (e.g., Old World Monkey, Ape, etc.) and human constant region sequences.

Monoclonal antibodies and antibody constructs include antibodies referred to as "human" or "fully human. " The terms "human antibody" and "fully human antibody" each refer to an antibody that has an amino acid sequence of a human immunoglobulin, including antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulins and that do not express endogenous immunoglobulins; for example, Xenomouse® antibodies and antibodies as described by<CIT>.

"Genetically altered antibodies" means antibodies wherein the amino acid sequence has been varied from that of a native antibody. Because of the relevance of recombinant DNA techniques in the generation of antibodies, one need not be confined to the sequences of amino acids found in natural antibodies; antibodies can be redesigned to obtain desired characteristics. The possible variations are many and range from changes to just one or a few amino acids to complete redesign of, for example, the variable and/or constant region. Changes in the constant region, in general, are made in order to improve or alter characteristics, such as complement fixation, interaction with membranes and other effector functions, as well as manufacturability and viscosity. Changes in the variable region can be made to improve antigen binding characteristics.

A "Fab fragment" is comprised of one light chain and the CH1 and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule.

A "Fab' fragment" contains one light chain and one heavy chain that contains more of the constant region, between the CH1 and CH2 domains, such that an interchain disulfide bond can be formed between two heavy chains to form a F(ab')<NUM> molecule.

A "F(ab')<NUM> fragment" contains two light chains and two heavy chains containing a portion of the constant region between the CH1 and CH2 domains, such that an interchain disulfide bond is formed between two heavy chains.

"Fv fragment" and "single chain antibody" refer to polypeptides containing antibody variable regions from both heavy and light chains but lacking constant regions. Like an intact antibody, an Fv fragment or single chain antibody are able to bind selectively to a specific antigen. With a molecular weight of only about <NUM> kDa, Fv fragments are much smaller than common antibodies (<NUM>-<NUM> kD), and even smaller than Fab fragments (about <NUM> kDa, one light chain and half a heavy chain).

A "single domain antibody" is an antibody fragment consisting of a single domain Fv unit, e.g., VH or VL. Like an intact antibody, a single domain antibody is able to bind selectively to a specific antigen. With a molecular weight of only <NUM>-<NUM> kDa, single-domain antibodies are much smaller than common antibodies (<NUM>-<NUM> kDa) which are composed of two heavy polypeptide chains and two light chains, and even smaller than Fab fragments (about <NUM> kDa, one light chain and half a heavy chain) and single-chain variable fragments (about <NUM> kDa, two variable domains, one from a light and one from a heavy chain). Nanobodies derived from light chains have also been shown to bind specifically to target epitopes.

"Amino acid" refers to either natural and/or unnatural or synthetic amino acids, including glycine and both the D and L optical isomers, amino acid analogs and peptidomimetics. In some aspects, the term amino acid refers to monomeric amino acids.

"Additive" means, in the context of a pharmaceutical composition, a substance not naturally part of a material (e.g., drug substance) but deliberately added to fulfill some specific purpose (e.g., preservation, viscosity reduction, stabilization).

"Surfactant" means surface-active agents, including substances commonly referred to as wetting agents, surface tension depressants, detergents, dispersing agents, emulsifiers, and quaternary ammonium antiseptics. Surfactants are further discussed below.

Proteins, including those that bind to one or more of the following, can be useful in the disclosed compositions and methods. These include CD proteins, including CD3, CD4, CD8, CD19, CD20, CD22, CD30, and CD34; including those that interfere with receptor binding. HER receptor family proteins, including HER2, HER3, HER4, and the EGF receptor. Cell adhesion molecules, for example, LFA-I, Mol, pl50, <NUM>, VLA-<NUM>, ICAM-I, VCAM, and alpha v/beta <NUM> integrin. Growth factors, such as vascular endothelial growth factor ("VEGF"), growth hormone, thyroid stimulating hormone, follicle stimulating hormone, luteinizing hormone, growth hormone releasing factor, parathyroid hormone, Mullerian-inhibiting substance, human macrophage inflammatory protein (MIP-I -alpha), erythropoietin (EPO), nerve growth factor, such as NGF-beta, platelet-derived growth factor (PDGF), fibroblast growth factors, including, for instance, aFGF and bFGF, epidermal growth factor (EGF), transforming growth factors (TGF), including, among others, TGF- α and TGF-β, including TGF-βI, TGF-β2, TGF-β3, TGF- β4, or TGF- β <NUM>, insulin-like growth factors-I and -II (IGF-I and IGF-II), des(I-<NUM>)-IGF-I (brain IGF-I), and osteoinductive factors. Insulins and insulin-related proteins, including insulin, insulin A-chain, insulin B-chain, proinsulin, and insulin-like growth factor binding proteins. Coagulation and coagulation-related proteins, such as, among others, factor VIII, tissue factor, von Willebrands factor, protein C, alpha-<NUM>-antitrypsin, plasminogen activators, such as urokinase and tissue plasminogen activator ("t-PA"), bombazine, thrombin, and thrombopoietin; (vii) other blood and serum proteins, including but not limited to albumin, IgE, and blood group antigens. Colony stimulating factors and receptors thereof, including the following, among others, M-CSF, GM-CSF, and G-CSF, and receptors thereof, such as CSF-<NUM> receptor (c-fms). Receptors and receptor-associated proteins, including, for example, flk2/flt3 receptor, obesity (OB) receptor, LDL receptor, growth hormone receptors, thrombopoietin receptors ("TPO-R," "c-mpl"), glucagon receptors, interleukin receptors, interferon receptors, T-cell receptors, stem cell factor receptors, such as c-Kit, and other receptors. Receptor ligands, including, for example, OX40L, the ligand for the OX40 receptor. Neurotrophic factors, including bone-derived neurotrophic factor (BDNF) and neurotrophin-<NUM>, -<NUM>, -<NUM>, or -<NUM> (NT-<NUM>, NT-<NUM>, NT-<NUM>, or NT-<NUM>). Relaxin A-chain, relaxin B-chain, and prorelaxin; interferons and interferon receptors, including for example, interferon-α, -β, and -y, and their receptors. Interleukins and interleukin receptors, including IL-I to IL-<NUM> and IL-I to IL-<NUM> receptors, such as the IL-<NUM> receptor, among others. Viral antigens, including an AIDS envelope viral antigen. Lipoproteins, calcitonin, glucagon, atrial natriuretic factor, lung surfactant, tumor necrosis factor-alpha and -beta, enkephalinase, RANTES (regulated on activation normally T-cell expressed and secreted), mouse gonadotropin-associated peptide, DNAse, inhibin, and activin. Integrin, protein A or D, rheumatoid factors, immunotoxins, bone morphogenetic protein (BMP), superoxide dismutase, surface membrane proteins, decay accelerating factor (DAF), AIDS envelope, transport proteins, homing receptors, addressins, regulatory proteins, immunoadhesins, antibodies. Myostatins, TALL proteins, including TALL-I, amyloid proteins, including but not limited to amyloid-beta proteins, thymic stromal lymphopoietins ("TSLP"), RANK ligand ("OPGL"), c-kit, TNF receptors, including TNF Receptor Type <NUM>, TRAIL-R2, angiopoietins, and biologically active fragments or analogs or variants of any of the foregoing.

Exemplary polypeptides and antibodies include Activase® (Alteplase); alirocumab, Aranesp® (Darbepoetin-alfa), Epogen® (Epoetin alfa, or erythropoietin); Avonex® (Interferon β-la); Bexxar® (Tositumomab); Betaseron® (Interferon-β); bococizumab (anti-PCSK9 monoclonal antibody designated as L1L3, see <CIT>); Campath® (Alemtuzumab); Dynepo® (Epoetin delta); Velcade® (bortezomib); MLN0002 (anti-α4β7 Ab); MLN1202 (anti-CCR2 chemokine receptor Ab); Enbrel® (etanercept); Eprex® (Epoetin alfa); Erbitux® (Cetuximab); evolocumab; Genotropin® (Somatropin); Herceptin® (Trastuzumab); Humatrope® (somatropin [rDNA origin] for injection); Humira® (Adalimumab); Infergen® (Interferon Alfacon-<NUM>); Natrecor® (nesiritide); Kineret® (Anakinra), Leukine® (Sargamostim); LymphoCide® (Epratuzumab); BenlystaTM (Belimumab); Metalyse® (Tenecteplase); Mircera® (methoxy polyethylene glycol-epoetin beta); Mylotarg® (Gemtuzumab ozogamicin); Raptiva® (efalizumab); Cimzia® (certolizumab pegol); SolirisTM (Eculizumab); Pexelizumab (Anti-C5 Complement); MEDI-<NUM> (Numax®); Lucentis® (Ranibizumab); Edrecolomab (,Panorex®); Trabio® (lerdelimumab); TheraCim hR3 (Nimotuzumab); Omnitarg (Pertuzumab, 2C4); Osidem® (IDM-I); OvaRex® (B43. <NUM>); Nuvion® (visilizumab); Cantuzumab mertansine (huC242-DMI); NeoRecormon® (Epoetin beta); Neumega® (Oprelvekin); Neulasta® (Pegylated filgastrim, pegylated G-CSF, pegylated hu-Met-G-CSF); Neupogen® (Filgrastim); Orthoclone OKT3® (Muromonab-CD3), Procrit® (Epoetin alfa); Remicade® (Infliximab), Reopro® (Abciximab), Actemra® (anti-IL6 Receptor Ab), Avastin® (Bevacizumab), HuMax-CD4 (zanolimumab), Rituxan® (Rituximab); Tarceva® (Erlotinib); Roferon-A®-(Interferon alfa-2a); Simulect® (Basiliximab); StelaraTM (Ustekinumab); Prexige® (lumiracoxib); Synagis® (Palivizumab); 146B7-CHO (anti-IL15 antibody, see <CIT>), Tysabri® (Natalizumab); Valortim® (MDX-<NUM>, anti-B. anthracis Protective Antigen Ab); ABthraxTM; Vectibix® (Panitumumab); Xolair® (Omalizumab), ETI211 (anti-MRSA Ab), IL-I Trap (the Fc portion of human IgGI and the extracellular domains of both IL-I receptor components (the Type I receptor and receptor accessory protein)), VEGF Trap (Ig domains of VEGFRI fused to IgGI Fc), Zenapax® (Daclizumab); Zenapax® (Daclizumab), Zevalin® (Ibritumomab tiuxetan), Zetia (ezetimibe), Atacicept (TACI-Ig), anti-α4β7 Ab (vedolizumab); galiximab (anti-CD80 monoclonal antibody), anti-CD23 Ab (lumiliximab); BR2-Fc (huBR3 / huFc fusion protein, soluble BAFF antagonist); SimponiTM (Golimumab); Mapatumumab (human anti-TRAIL Receptor-<NUM> Ab); Ocrelizumab (anti-CD20 human Ab); HuMax-EGFR (zalutumumab); M200 (Volociximab, anti-α5β1 integrin Ab); MDX-<NUM> (Ipilimumab, anti-CTLA-<NUM> Ab and VEGFR-I (IMC-18F1); anti-BR3 Ab; anti-C. difficile Toxin A and Toxin B C Abs MDX-<NUM> (CDA-I) and MDX-<NUM>); anti-CD22 dsFv-PE38 conjugates (CAT-<NUM> and CAT-<NUM>); anti-CD25 Ab (HuMax-TAC); anti-TSLP antibodies; anti-TSLP receptor antibody (see <CIT>); anti-TSLP antibody designated as A5 (see <CIT>); (see anti-CD3 Ab (NI-<NUM>); Adecatumumab (MT201, anti-EpCAM-CD326 Ab); MDX-<NUM>, SGN-<NUM>, SGN-<NUM> (anti-CD30 Abs); MDX-<NUM> (anti- IFNAR); HuMax CD38 (anti-CD38 Ab); anti-CD40L Ab; anti-Cripto Ab; anti-CTGF Idiopathic Pulmonary Fibrosis Phase I Fibrogen (FG-<NUM>); anti-CTLA4 Ab; anti-eotaxinl Ab (CAT-<NUM>); anti-FGF8 Ab; anti-ganglioside GD2 Ab; anti-sclerostin antibodies (see, <CIT> or <CIT>) anti-sclerostin antibody designated as Ab-<NUM> (see <CIT> or <CIT>); anti-ganglioside GM2 Ab; anti-GDF-<NUM> human Ab (MYO-<NUM>); anti-GM-CSF Receptor Ab (CAM-<NUM>); anti-HepC Ab (HuMax HepC); MEDI-<NUM>, MDX-<NUM> (anti-IFNα Ab); anti-IGFIR Ab; anti-IGF-IR Ab (HuMax-Inflam); anti-IL12/IL23p40 Ab (Briakinumab); anti-IL-23p19 Ab (LY2525623); anti-IL13 Ab (CAT-<NUM>); anti-IL-<NUM> Ab (AIN457); anti-IL2Ra Ab (HuMax-TAC); anti-IL5 Receptor Ab; anti-integrin receptors Ab (MDX-OI8, CNTO <NUM>); anti-IPIO Ulcerative Colitis Ab (MDX- <NUM>); anti-LLY antibody; BMS-<NUM>; anti-Mannose Receptor/hCGβ Ab (MDX-<NUM>); anti-mesothelin dsFv-PE38 conjugate (CAT-<NUM>); anti-PDIAb (MDX-<NUM><NUM> (ONO- <NUM>)); anti-PDGFRα antibody (IMC-3G3); anti-TGFβ Ab (GC-<NUM>); anti-TRAIL Receptor-<NUM> human Ab (HGS-ETR2); anti-TWEAK Ab; anti-VEGFR/Flt-<NUM> Ab; anti- ZP3 Ab (HuMax-ZP3); NVS Antibody #<NUM>; NVS Antibody #<NUM>; and an amyloid-beta monoclonal antibody comprising sequences, SEQ ID NO:<NUM> and SEQ ID NO:<NUM> (see <CIT>).

Examples of antibodies suitable for the methods and pharmaceutical formulations include the antibodies shown in Table <NUM>. Other examples of suitable antibodies include infliximab, bevacizumab, ranibizumab, cetuximab, ranibizumab, palivizumab, abagovomab, abciximab, actoxumab, adalimumab, afelimomab, afutuzumab, alacizumab, alacizumab pegol, ald518, alemtuzumab, alirocumab, alemtuzumab, altumomab, amatuximab, anatumomab mafenatox, anrukinzumab, apolizumab, arcitumomab, aselizumab, altinumab, atlizumab, atorolimiumab, tocilizumab, bapineuzumab, basiliximab, bavituximab, bectumomab, belimumab, benralizumab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab, biciromab, bivatuzumab, bivatuzumab mertansine, blinatumomab, blosozumab, brentuximab vedotin, briakinumab, brodalumab, canakinumab, cantuzumab mertansine, cantuzumab mertansine, caplacizumab, capromab pendetide, carlumab, catumaxomab, cc49, cedelizumab, certolizumab pegol, cetuximab, citatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan, conatumumab, crenezumab, cr6261, dacetuzumab, daclizumab, dalotuzumab, daratumumab, demcizumab, denosumab, detumomab, dorlimomab aritox, drozitumab, duligotumab, dupilumab, ecromeximab, eculizumab, edobacomab, edrecolomab, efalizumab, efungumab, elotuzumab, elsilimomab, enavatuzumab, enlimomab pegol, enokizumab, enokizumab, enoticumab, enoticumab, ensituximab, epitumomab cituxetan, epratuzumab, erlizumab, ertumaxomab, etaracizumab, etrolizumab, exbivirumab, exbivirumab, fanolesomab, faralimomab, farletuzumab, fasinumab, fbta05, felvizumab, fezakinumab, ficlatuzumab, figitumumab, flanvotumab, fontolizumab, foralumab, foravirumab, fresolimumab, fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab, gemtuzumab ozogamicin, gevokizumab, girentuximab, glembatumumab vedotin, golimumab, gomiliximab, gs6624, ibalizumab, ibritumomab tiuxetan, icrucumab, igovomab, imciromab, imgatuzumab, inclacumab, indatuximab ravtansine, infliximab, intetumumab, inolimomab, inotuzumab ozogamicin, ipilimumab, iratumumab, itolizumab, ixekizumab, keliximab, labetuzumab, lebrikizumab, lemalesomab, lerdelimumab, lexatumumab, libivirumab, ligelizumab, lintuzumab, lirilumab, lorvotuzumab mertansine, lucatumumab, lumiliximab, mapatumumab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minretumomab, mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomab pasudotox, muromonab-cd3, nacolomab tafenatox, namilumab, naptumomab estafenatox, narnatumab, natalizumab, nebacumab, necitumumab, nerelimomab, nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan, ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab, olokizumab, omalizumab, onartuzumab, oportuzumab monatox, oregovomab, orticumab, otelixizumab, oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, panitumumab, panobacumab, parsatuzumab, pascolizumab, pateclizumab, patritumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pintumomab, placulumab, ponezumab, priliximab, pritumumab, PRO <NUM>, quilizumab, racotumomab, radretumab, rafivirumab, ramucirumab, ranibizumab, raxibacumab, regavirumab, reslizumab, rilotumumab, rituximab, robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab, ruplizumab, samalizumab, sarilumab, satumomab pendetide, secukinumab, sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab, siplizumab, sirukumab, solanezumab, solitomab, sonepcizumab, sontuzumab, stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tanezumab, taplitumomab paptox, tefibazumab, telimomab aritox, tenatumomab, tefibazumab, telimomab aritox, tenatumomab, teneliximab, teplizumab, teprotumumab, TGN1412, tremelimumab, ticilimumab, tildrakizumab, tigatuzumab, TNX-<NUM>, tocilizumab, toralizumab, tositumomab, tralokinumab, trastuzumab, TRBS07, tregalizumab, tremelimumab, tucotuzumab celmoleukin, tuvirumab, ublituximab, urelumab, urtoxazumab, ustekinumab, vapaliximab, vatelizumab, vedolizumab, veltuzumab, vepalimomab, vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab, zalutumumab, zanolimumab, zatuximab, ziralimumab and zolimomab aritox.

Most preferred antibodies for use in the disclosed formulations and methods are adalimumab, bevacizumab, blinatumomab, cetuximab, conatumumab, denosumab, eculizumab, erenumab, evolocumab, infliximab, natalizumab, panitumumab, rilotumumab, rituximab, romosozumab, and trastuzumab, and antibodies selected from Table <NUM>.

Exemplary polypeptide concentrations in the formulation may range from about <NUM>/ml to about <NUM>/ml (or more), about <NUM>/ml to about <NUM>/ml, from about <NUM>/ml to about <NUM>/ml, from about <NUM>/ml to about <NUM>/ml, or from about <NUM>/ml to about <NUM>/ml, or alternatively from about <NUM>/ml to about <NUM>/ml, such as <NUM>/ml. The concentration of protein will depend upon the end use of the pharmaceutical formulation and can be easily determined by a person of skill in the art. Particularly contemplated concentrations of protein are at least <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM> and <NUM>/ml and including all values in between.

Acceptable formulation components preferably are nontoxic to patients at the dosages and concentrations used. Pharmaceutical formulations can comprise agents for modifying, maintaining or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition.

In general, excipients can be classified on the basis of the mechanisms by which they stabilize proteins against various chemical and physical stresses. Some excipients alleviate the effects of a specific stress or regulate a particular susceptibility of a specific polypeptide. Other excipients more generally affect the physical and covalent stabilities of proteins.

Common excipients of liquid and lyophilized protein formulations are shown in Table <NUM> (see also (Kamerzell, Esfandiary, Joshi, Middaugh, & Volkin, <NUM>)).

Other excipients are known in the art (e.g., see (Powell, Nguyen, & Baloian, <NUM>)).

Those skilled in the art can determine what amount or range of excipient can be included in any particular formulation to achieve a biopharmaceutical formulation that promotes retention in stability of the biopharmaceutical. For example, the amount and type of a salt to be included in a biopharmaceutical formulation can be selected based on to the desired osmolality (i.e., isotonic, hypotonic or hypertonic) of the final solution as well as the amounts and osmolality of other components to be included in the formulation.

Solution pH affects the chemical integrity of a polypeptide's amino acid residues (e.g., Asn deamidation and Met oxidation) and maintenance of its higher order structure. Buffering agents are used to control solution pH and optimize protein stability. Maximal stability of a polypeptide drug is often within a narrow pH range. Several approaches (e.g., accelerated stability studies and calorimetric screening studies) are useful for this purpose. Once a formulation is finalized, the drug product must be manufactured and maintained within a predefined specification throughout its shelf-life. Hence, buffering agents are almost always used to control pH in the formulation.

Organic acids, phosphates and Tris can be used as buffers in polypeptide formulations (see Table <NUM>). The buffer capacity of the buffering species is maximal at a pH equal to the pKa and decreases as pH increases or decreases away from this value. Ninety percent of the buffering capacity exists within one pH unit of its pKa. Buffer capacity also increases proportionally with increasing buffer concentration.

In addition to the foregoing, some therapeutic polypeptides can be "self-buffering" at a pharmaceutically sufficient concentration. Formulations of such polypeptides can often dispense with a conventional buffer.

A pH buffering compound can be present in any amount suitable to maintain the pH of the formulation at a predetermined level. When the pH buffering agent is an amino acid, for example, the concentration of the amino acid is often between <NUM> and <NUM> (<NUM>). The pH buffering agent can be at least <NUM>, <NUM>, <NUM>, <NUM><NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>. In some cases, the concentration of the pH buffering agent is between <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> and <NUM>. In other instances, the concentration of the pH buffering agent is between <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> and <NUM>. For example, the pH buffering agent is <NUM>. In some case, the buffer is <NUM> phosphate. In other examples, the buffer is <NUM> acetate.

Sugars are frequently used to stabilize polypeptides in both liquid and lyophilized formulations. Disaccharides, such as sucrose and trehalose, are thought to stabilize proteins by preferential hydration at high concentrations in the liquid state and by specific interactions with polypeptides and formation of viscous glassy matrices in the solid state. Sugar molecules can increase the viscosity of monoclonal antibody solutions, presumably due to a preferential hydration mechanism. Sugar alcohols, such as sorbitol, can stabilize polypeptides in solution and in the lyophilized state. Mannitol is often used as a bulking agent in lyophilized formulations. Lactose can be used as a carrier molecule for inhaled formulations of polypeptides. Cyclodextrin derivatives can stabilize proteins in liquid formulations of antibodies, vaccine antigens, and such smaller proteins as growth factors, interleukin-<NUM> and insulin.

Bulking agents are typically used in lyophilized formulations to enhance product elegance and to prevent blowout. Conditions in the formulation are generally designed so that the bulking agent crystallizes out of the frozen amorphous phase (either during freezing or annealing above the glass transition temperature of maximally freeze-concentrated solutes (Tg')) giving the cake structure and bulk. Mannitol and glycine are examples of commonly used bulking agents.

Stabilizers include compounds that can serve as cryoprotectants, lyoprotectants, and glass-forming agents. Cryoprotectants act to stabilize polypeptides during freezing or in the frozen state at low temperatures. Lyoprotectants stabilize polypeptides in the freeze-dried solid dosage form by preserving the native-like conformational properties of the protein during dehydration stages of freeze-drying. Glassy state properties have been classified as "strong" or "fragile" depending on their relaxation properties as a function of temperature. It is important that cryoprotectants, lyoprotectants, and glass-forming agents remain in the same phase with the polypeptide in order to impart stability. Sugars, polymers, and polyols fall into this category and can sometimes serve all three roles of cryoprotectants, lyoprotectans, and glass-forming agents.

Polyols encompass a class of excipients that includes sugars, (e.g. mannitol, sucrose, sorbitol), and other polyhydric alcohols (e.g., glycerol and propylene glycol). The polymer polyethylene glycol (PEG) is included in this category. Polyols are commonly used as stabilizing excipients and/or isotonicity agents in both liquid and lyophilized parenteral polypeptide formulations. With respect to the Hofmeister series, the polyols are kosmotropic and are preferentially excluded from the polypeptide surface. Polyols can protect polypeptides from both physical and chemical degradation. Preferentially excluded co-solvents increase the effective surface tension of solvent at the polypeptide interface whereby the most energetically favorable polypeptide conformations are those with the smallest surface areas.

Mannitol is often used as a bulking agent in lyophilized formulations because it crystallizes out of the amorphous protein phase during freeze-drying lending structural stability to the cake (e.g., Leukine ®, Enbrel ® - Lyo, Betaseron®). It is generally used in combination with a cryo and/or lyoprotectant, like sucrose. Because of the propensity of mannitol to crystallize under frozen conditions, sorbitol and sucrose are preferred tonicity agents/stabilizers in liquid formulations to protect the product against freeze-thaw stresses encountered during transport or when freezing bulk prior to manufacturing. Sorbitol and sucrose are far more resistant to crystallization and therefore less likely to phase separate from the polypeptide. The use of reducing sugars containing free aldehyde or ketone groups, such as glucose and lactose, is preferably avoided because they can react and glycate surface lysine and arginine residues of polypeptides via the Maillard reaction of aldehydes and primary amines. Sucrose can hydrolyze to fructose and glucose under acidic conditions, and consequently may cause glycation.

A stabilizer (or a combination of stabilizers) can be added to a lyophilization formulation to prevent or reduce lyophilization-induced or storage-induced aggregation and chemical degradation. A hazy or turbid solution upon reconstitution indicates that the polypeptide has precipitated. "Stabilizer" means an excipient capable of preventing aggregation or other physical degradation, as well as chemical degradation (for example, autolysis, deamidation, oxidation, etc.) in an aqueous and solid state. Stabilizers that are conventionally used in pharmaceutical compositions include sucrose, trehalose, mannose, maltose, lactose, glucose, raffinose, cellobiose, gentiobiose, isomaltose, arabinose, glucosamine, fructose, mannitol, sorbitol, glycine, arginine HCl, poly-hydroxy compounds, including polysaccharides such as dextran; starch, hydroxyethyl starch, cyclodextrins, N-methyl pyrollidene, cellulose and hyaluronic acid, sodium chloride.

Examples of osmolytes are presented in Table A. Other osmolytes that can be useful as excipients include taurine, betaine, trimethylamine N-oxide (TMAO), choline-O-sulfate, and sarcosine.

Pharmaceutical formulation are preferably isotonic, with an osmolality ranging from between about <NUM> to about <NUM> mOsm/kg, e.g., about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, about <NUM> mOsm/kg, or about <NUM> mOsm/kg. Osmolality is the measure of the ratio of solutes to volume fluid. In other words, it is the number of molecules and ions (or molecules) per kilogram of a solution. The osmolality can be <NUM> mOsm/kg. Osmolality can be measured by an osmometer, such as Advanced Instruments <NUM> Multi-sample Osmometer, Norwood, MA. The Advanced Instruments <NUM> Multi-sample Osmometer measures osmolality by using the Freezing Point Depression method. The higher the osmolytes in a solution, the temperature in which it will freeze drops. Osmolality can also be measured using any other methods and in any other units known in the art such as linear extrapolation. The pharmaceutical formulation can be isotonic to a human blood cell, such as a red blood cell.

Polypeptide-based excipients add complexity to the formulation, especially in developing analytical methods to monitor the stability of the polypeptide-based drug or vaccine in the presence of a polypeptide-based excipient. Polymers have been evaluated as excipients (e.g., as bulking agents) in lyophilized polypeptide formulations. Controlled release formulations of polypeptide drugs and vaccines in which polypeptides are formulated with polymers, such as PLGA (poly(lactic-co-glycolic acid) and PEG (polyethylene glycol), can also be made. Many additional water-soluble polymers (e.g., HEC (hydroxyethylcellulose), CMC (carboxymethyl cellulose) can be used to formulate polypeptide drugs for topical application.

Reducing agents, oxygen/free-radical scavengers, and chelating agents and be used as antioxidants in pharmaceutical formulations. Antioxidants in therapeutic polypeptide formulations must be water-soluble and remain active throughout the product shelf-life. Reducing agents and oxygen/free-radical scavengers work by ablating active oxygen species in solution. Chelating agents (e.g., EDTA (ethylenediamine tetra-acetic acid)) can be effective by binding trace metal contaminants that promote free-radical formation. In the liquid formulation of acidic fibroblast growth factor, for example, EDTA inhibits metal ion-catalyzed oxidation of cysteine residues.

In general, transition metal ions are undesired in polypeptide formulations because they can catalyze physical and chemical degradation reactions in polypeptide drug products. Specific metal ions are included in formulations, however, when they act as co-factors to polypeptides. Metal ions can also be used in suspension formulations of polypeptides where they form coordination complexes (e.g., zinc suspensions of insulin). Magnesium ions (<NUM>-<NUM>) can be usedto inhibit the isomerization of aspartic acid to isoaspartic acid.

One approach to improve the conformational stability of polypeptide therapeutic drugs is to take advantage of the polypeptide's inherent ligand binding sites. For example, Pulmozyme® not only requires bivalent metal ions for its enzymatic activity, but it has improved conformational stability in the presence of calcium ions. Both acidic and basic fibroblast growth factors (aFGF and bFGF) naturally bind to the highly negatively charged proteoglycans on cell surfaces. A variety of other highly negatively charged compounds also bind and dramatically stabilize aFGF by interaction with the protein's polyanion binding site.

Polypeptide molecules have a high propensity to interact with surfaces, making them susceptible to adsorption and denaturation at air-liquid, vial-liquid, and liquid-liquid (silicone oil) interfaces. This phenomenon is inversely dependent on polypeptide concentration and results in soluble or insoluble polypeptide aggregates or the loss of polypeptide from solution through surface adsorption. In addition to container surface adsorption, surface-induced degradation is exacerbated with physical agitation, as can be experienced during shipping and handling.

Surfactants are commonly used in polypeptide formulations to prevent surface-induced degradation. Surfactants are amphipathic molecules with the capability of out-competing polypeptides for interfacial positions. Hydrophobic portions of surfactant molecules occupy interfacial positions (e.g., air/liquid), while hydrophilic portions of surfactant molecules remain oriented towards the bulk solvent. At sufficient concentrations (typically around the detergent's critical micellar concentration), a surface layer of surfactant molecules serve to prevent protein molecules from adsorbing at the interface, minimizing surface-induced degradation.

The most commonly used surfactants are the non-ionic fatty acid esters of sorbitan polyethoxylates--i.e., polysorbate <NUM> and polysorbate <NUM> (e.g., found in Avonex®, Neupogen®, Neulasta®). The two differ only in the length of the aliphatic chain that imparts hydrophobic character to the molecules, C-<NUM> and C-<NUM>, respectively. Polysorbate <NUM> is more surface-active and has a lower critical micellar concentration than polysorbate <NUM>. Both polysorbate <NUM> and polysorbate <NUM> have been shown to protect against agitation-induced aggregation. Polysorbate <NUM> and <NUM> also protect against stress induced by freezing, lyophilization and reconstitution. The surfactant poloxamer <NUM> has also been used in several marketed liquid products, such Gonal-F®, Norditropin®, and Ovidrel®. Non-ionic surfactants stabilize polypeptides primarily by outcompeting polypeptide molecules for hydrophobic surfaces (e.g., air-water interfaces), thereby preventing polypeptides from unfolding at these hydrophobic interfaces. Non-ionic surfactants can also block polypeptide molecules from adsorbing to other hydrophobic surfaces present during processing. In addition, non-ionic surfactants can directly interact with hydrophobic regions in polypeptide molecules.

Other examples of surfactants include polyoxyethylene-polyoxypropylene block copolymer (Poloxamers such as Pluronic® F-<NUM> and other Pluronics®), other sorbitan alkyl esters (Spans®), Polyethylene glycol alkyl ethers (Brij), Polypropylene glycol alkyl ethers, Glucoside alkyl ethers, and D-α-tocopherol polyethylene glycol succinate (vitamin E TPGS).

Surfactants can also affect the thermodynamic conformational stability of polypeptides. The effects of a given excipient are polypeptide-specific. For example, polysorbates can reduce the stability of some polypeptides and increase the stability of others. Surfactant destabilization of polypeptides can be rationalized in terms of the hydrophobic tails of the detergent molecules that can engage in specific binding with partially or wholly unfolded polypeptide states. These types of interactions can cause a shift in the conformational equilibrium towards the more expanded polypeptide states (i.e., increasing the exposure of hydrophobic portions of the polypeptide molecule in complement to binding polysorbate). Alternatively, if the polypeptide native state exhibits some hydrophobic surfaces, detergent binding to the native state can stabilize that conformation.

For surfactants, the effective concentration for a given polypeptide depends on the mechanism of stabilization.

Surfactants can also be added in appropriate amounts to prevent surface-related aggregation during freezing and drying. Exemplary surfactants include anionic, cationic, nonionic, zwitterionic, and amphoteric surfactants, including surfactants derived from naturally occurring amino acids. Anionic surfactants include sodium lauryl sulfate (SDS), dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate, chenodeoxycholic acid, N-lauroylsarcosine sodium salt, lithium dodecyl sulfate, <NUM>-octanesulfonic acid sodium salt, sodium cholate hydrate, sodium deoxycholate, and glycodeoxycholic acid sodium salt. Cationic surfactants include benzalkonium chloride or benzethonium chloride, cetylpyridinium chloride monohydrate, and hexadecyltrimethylammonium bromide. Zwitterionic surfactants include CHAPS, CHAPSO, SB3-<NUM>, and SB3-<NUM>. Non-ionic surfactants include digitonin, TRITON™ X-<NUM>, TRITON™ X-<NUM>, TWEEN®-<NUM>, and TWEEN®-<NUM>. Surfactants also include lauromacrogol <NUM>, polyoxyl <NUM> stearate, polyoxyethylene hydrogenated castor oil <NUM>, <NUM>, <NUM> and <NUM>, glycerol monostearate, polysorbate <NUM>, <NUM>, <NUM> and <NUM>, soy lecithin and other phospholipids, such as <NUM>,<NUM>-Dioleoyl-sn-glycero-<NUM>-phosphocholine (DOPC), <NUM>,<NUM>-Dimyristoyl-sn-glycero-<NUM>-phospho-rac-(<NUM>-glycerol) sodium salt (DMPG), <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine (DMPC), and <NUM>,<NUM>-Dioleoyl-sn-glycero-<NUM>-phospho-rac-(<NUM>-glycerol) sodium salt (DOPG); sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. The surfactant can be in a concentration of about <NUM>% to about <NUM>% w/v, such as in a concentration of at least about <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>% w/v. In another example, the surfactant is incorporated in a concentration of about <NUM>% to about <NUM>% w/v. In still another example, the surfactant is incorporated in a concentration of about <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>% w/v to about <NUM>% w/v. In yet another example, the surfactant is incorporated in a concentration of about <NUM>% to about <NUM>% w/v.

Salts are often added to increase the ionic strength of the formulation, which can be important for polypeptide solubility, physical stability, and isotonicity. Salts can affect the physical stability of polypeptides in a variety of ways. Ions can stabilize the native state of polypeptides by binding to charged residues on the polypeptide's surface. Alternatively, they can stabilize the denatured state by binding to the peptide groups along the polypeptide backbone. Salts can also stabilize the polypeptide native conformation by shielding repulsive electrostatic interactions between residues within a polypeptide. Electrolytes in polypeptide formulations can also shield attractive electrostatic interactions between polypeptide molecules that can lead to protein aggregation and insolubility.

The effect of salt on the stability and solubility of polypeptides varies significantly with the characteristics of the ionic species. The Hofmeister series originated in the <NUM>'s as a way to rank order electrolytes based on their ability to precipitate polypeptides. The Hofmeister series can used to illustrate polypeptide stabilization effects by ionic and non-ionic co-solutes, as shown in Table <NUM> (Cacace, Landau, & Ramsden, <NUM>). In general, the differences in effects across the anions are far greater than that observed for the cations, and, for both types, the effects are most apparent at higher concentrations than are acceptable in parenteral formulations. High concentrations of kosmotropes (e.g., ><NUM> molar ammonium sulfate) are commonly used to precipitate polypeptides from solution (salting-out) where the kosmotrope is preferentially excluded from the polypeptide surface reducing the solubility of the polypeptide in its native conformation. Removal or dilution of the salt returns the polypeptide to solution. Salting in occurs when destabilizing ions are used to increase the solubility of polypeptides by solvating the peptide bonds of the polypeptide backbone. Increasing concentrations of the chaotrope favor the denatured state of the polypeptide as the solubility of the peptide chain increases. The relative effectiveness of ions to salt-in and salt-out defines their position in the Hofmeister series.

In order to maintain isotonicity in a parenteral formulation, salt concentrations are generally limited to less than <NUM> for monovalent ion combinations. In this concentration range, the mechanism of salt stabilization is probably due to screening of electrostatic repulsive intramolecular forces or attractive intermolecular forces (Debye-Huckel screening). Interestingly, chaotropic salts can be more effective at stabilizing polypeptide structure than similar concentrations of kosmotropes by this mechanism. The chaotropic anions bind more strongly than the kosmotropic ions. With respect to covalent polypeptide degradation, differential effects of ionic strength on this mechanism are expected through Debye-Huckel theory. The mechanisms by which salts affect polypeptide stability are polypeptide-specific and can vary significantly as a function of solution pH.

Preservatives can be necessary when developing multi-use parenteral formulations that involve more than one extraction from the same container. Their primary function is to inhibit microbial growth and ensure product sterility throughout the shelf-life or term of use of the drug product. Commonly used preservatives include benzyl alcohol, phenol and m-cresol. Development of polypeptide formulations that include preservatives can be challenging. Preservatives almost always have a destabilizing effect (aggregation) on polypeptides. Benzyl alcohol has also been shown to affect polypeptide structure and stability in a concentration-, temperature- and time-dependent manner. Due to these destabilizing effects, many lyophilized polypeptide formulations are reconstituted with diluent containing benzyl alcohol to minimize the contact time with the polypeptide prior to administration.

Several aspects need to be considered during the formulation development of preserved dosage forms. The effective preservative concentration in the drug product must be optimized. This requires testing a given preservative in the dosage form with concentration ranges that confer anti-microbial effectiveness without compromising polypeptide stability.

Development of liquid formulations containing preservatives are often more challenging than lyophilized formulations. Freeze-dried products can be lyophilized without a preservative and reconstituted with a preservative containing diluent at the time of use. With liquid formulations, preservative effectiveness and stability have to be maintained over the entire product shelf-life (usually about <NUM> -<NUM> months). Preservative effectiveness often needs to be demonstrated in the final formulation containing the active drug and all excipient components.

Some preservatives can cause injection site reactions. For example, patient pain perception can be lower in formulations containing phenol and benzyl alcohol as compared to formulations containing m-cresol. Benzyl alcohol appears to possess anesthetic properties.

Reducing the viscosity of therapeutic polypeptide formulations is of interest in the pharmaceutical arts. The dipeptide excipients N-acetyl-serine-arginine N-acetyl-proline-arginine, and N-acetyl-proline-arginine-NH<NUM> were discovered to reduce the viscosity of therapeutic antibody formulations. Provided herein are viscosity-reducing excipients at selected concentrations for use in reducing the viscosity of therapeutic polypeptide (such as therapeutic antibodies) formulations. Provided herein are therapeutic polypeptide and antibody formulations and methods for reducing the viscosity of therapeutic polypeptide and antibody formulations by combining the therapeutic polypeptide or antibody with a viscosity-reducing concentration of a N-acetyl-serine-arginine, N-acetyl-proline-arginine and/or N-acetyl-proline-NH<NUM>-arginine dipeptides.

N-acetyl-serine-arginine, N-acetyl-proline-arginine, and N-acetyl-proline-arginine-NH<NUM> dipeptides can be synthesized by understood methods in the art, such as by solid-state peptide synthesis. It is advantageous however, to use trifluoroacetic acid (TFA)-free methods, such as those that use, for example, HCl. In such methods, it can be advantageous to facilitate purification of the synthesized N-acetyl-dipeptides by purifying the protected dipeptides before deprotection.

The concentration of N-acetyl-serine-arginine, N-acetyl-proline-arginine, and N-acetyl-proline-arginine-NH<NUM> can be experimentally determined by one of ordinary skill. In some examples, the N-acetyl-dipeptide can have a concentration from about <NUM> to about <NUM>, such as from about <NUM> to about <NUM>, <NUM>, <NUM>, and about <NUM>. For example, the N-acetyl-dipeptide can be present from about (in mM) <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, and <NUM>.

In one aspect, the disclosed pharmaceutical formulations have a viscosity level of less than about <NUM> centipoise (cP) as measured at room temperature (i.e., <NUM>). The pharmaceutical formulation can have a viscosity level of less than about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP; about <NUM> cP, about <NUM> cP, about <NUM> cP; about <NUM> cP; or about <NUM> cP. Since <NUM> cP = <NUM> mPa·s, values referred to in cP equal the same values in mPa·s.

In one aspect, the pharmaceutical formulation is stable as measured by at least one stability assay, such as an assay that examines the biophysical or biochemical characteristics of the antibody over time. Pharmaceutical formulation stability can be measured using SEC-HPLC. SEC-HPLC separates proteins based on differences in their hydrodynamic volumes. Molecules with larger hydrodynamic proteins volumes elute earlier than molecules with smaller volumes. In the case of SEC-HPLC, a stable pharmaceutical formulation exhibits no more than about a <NUM>% increase in HMW species as compared to a control sample, such as, for example no more than about a <NUM>%, no more than about a <NUM>%, no more than about a <NUM>%, no more than about a <NUM>%, no more than about a <NUM>% increase in HMW species as compared to a control sample.

Alternatively, or in addition, stability can be measured using cation-exchange HPLC (CEX-HPLC). CEX-HPLC separates proteins based on differences in their surface charge. At a set pH, charged isoforms of an antibody are separated on a cation-exchange column and eluted using a salt gradient. The eluent is monitored by ultraviolet light (UV) absorbance. The charged isoform distribution is evaluated by determining the peak area of each isoform as a percent of the total peak area. In the case of CEX-HPLC, a stable pharmaceutical formulation exhibits no more than about a <NUM>% decrease in the main isoform peak as compared to a control sample, such as, for example, no more than about a <NUM>% to about a <NUM>% decrease in the main isoform peak as compared to a control sample; no more than about a <NUM>% decrease, no more than about a <NUM>% decrease, no more than about a <NUM>% decrease, no more than about a <NUM>% decrease, no more than about a <NUM>% decrease in the main isoform peak as compared to a control sample.

Also alternatively, or in addition, formulation stability can be measured using Subvisible Particle Detection by Light Obscuration (HIAC). An electronic, liquid-borne particle-counting system (HIAC/Royco <NUM> (Hach Company; Loveland, CO) or equivalent) containing a light-obscuration sensor (HIAC/Royco HRLD-<NUM> or equivalent) with a liquid sampler quantifies the number of particles and their size range in a given test sample. When particles in a liquid pass between the light source and the detector they diminish or "obscure" the beam of light that falls on the detector. When the concentration of particles lies within the normal range of the sensor, these particles are detected one-by-one. The passage of each particle through the detection zone reduces the incident light on the photo-detector and the voltage output of the photo-detector is momentarily reduced. The changes in the voltage register as electrical pulses that are converted by the instrument into the number of particles present. The method is nonspecific and measures particles regardless of their origin. Particle sizes monitored are generally <NUM>, and <NUM>. In the case of HIAC, a stable pharmaceutical formulation exhibits no more than <NUM><NUM> particles per container (or unit), as compared to a control sample, such as, for example no more than <NUM>, no more than <NUM>, no more than <NUM>, no more than <NUM>, no more than <NUM>, <NUM> particles per container (or unit) as compared to a control sample. In other cases, a stable pharmaceutical formulation exhibits no more than <NUM><NUM> particles per container (or unit) as compared to a control sample, such as, for example, no more than <NUM>, no more than <NUM>, no more than <NUM>, no more than <NUM>, no more than <NUM>, no more than <NUM><NUM> particles per container (or unit) as compared to a control sample.

Pharmaceutical formulation stability can also be assessed using visual assessment. Visual assessment is a qualitative method used to describe the visible physical characteristics of a sample. The sample is viewed against a black and/or white background of an inspection booth, depending on the characteristic being evaluated (e.g., color, clarity, presence of particles or foreign matter). Samples are also viewed against an opalescent reference standard and color reference standards. In the case of visual assessment, a stable pharmaceutical formulation exhibits no significant change in color, clarity, presence of particles or foreign matter as compared to a control sample.

Pharmaceutical formulations disclosed herein can be prepared by either of two processes designated processes <NUM> and <NUM>. Process <NUM> comprises:.

In process <NUM>, the pH of the concentrated protein to achieve the desired final pH can range from about <NUM> to about <NUM>. In process <NUM>, the pH of the concentrated protein solution to achieve the desired final pH can range from about <NUM> to about <NUM>. Where a particular excipient is reported in a formulation by, for example, percent (%) w/v, those skilled in the art recognize that the equivalent molar concentration of that excipient is also contemplated.

Once the pharmaceutical formulation has been formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or as a dehydrated or lyophilized powder. Such formulations can be stored either in a ready-to-use form or in a form (e.g., lyophilized) that is reconstituted prior to administration. In some cases, the therapeutic polypeptide formulations can be stored in containers, such as suitable storage bags (e.g., as manufactured by Sartorius (Gottingen, DE)) or in polycarbonate carboys. Once the pharmaceutical formulation has been formulated, it can also be stored in pre-filled syringes (PFS; such as <NUM> PFS's) as a solution or suspension in a ready-to-use form, as well as in glass vials (such as <NUM> cc glass vials).

Kits may be provided for producing a single-dose administration unit. The kit can contain both a first container having a dried protein and a second container having an aqueous formulation. Kits containing single and multi-chambered pre-filled syringes (e.g., liquid syringes and lyosyringes) can be included.

The following Examples section is given solely by way of example.

N-acetyl-arginine and N-acetyl-arginine-NH<NUM> were purchased from BACHEM (Torrance, CA). arginine HCl was purchased from SAFC Ajinomoto (Sigma-Aldrich, St. Louis, MO; Itasca, IL). N-acetyl-proline-arginine dipeptide, N-acetyl-proline-arginine-NH<NUM>, and N-acetyl-serine-arginine dipeptide were purchased from AnaSpec (Fremont, CA). The glutamate-arginine dipeptide was also sourced from AnaSpec. A <NUM>% wt polysorbate <NUM> stock solution at pH <NUM> was prepared using glacial acetic acid and 2N NaOH was used for titration. This stock solution also served as a control. The remaining five formulations were prepared to contain <NUM> of each excipient. The glutamate-arginine dipeptide formulation was used as a reference.

The protein concentration of each sample was measured using SoloVPE UV (C Technologies; Bridgewater, NJ) spectroscopy. The samples were stored at <NUM>-<NUM> until being brought to room temperature prior to sample loading on the viscometer. The samples were measured within <NUM> weeks of preparation (usually within <NUM>-<NUM> days).

The viscosity of the protein formulations was measured using a standard cone-and-plate rotational viscometer (AR-G2 TA Instruments (New Castle, DE) using a <NUM> diameter with <NUM> degree cone) at a temperature <NUM> and a shear rate range of <NUM>-<NUM>-<NUM>). Upon loading, each sample was equilibrated for <NUM> minutes at <NUM> prior to the start of data collection. All formulation samples tested showed Newtonian rheological behavior. Therefore, the viscosity values reported herein were average values at a shear rate range of <NUM>-<NUM>-<NUM>.

The objective of this example was to determine the ability of N-acetyl-proline-arginine and N-acetyl-serine-arginine dipeptides to reduce the viscosity of a high concentration therapeutic Ab. N-acetyl-arginine (NAR, see for example Sloey and Kanapuram (<NUM>)) was used for comparison, as was arginine.

Ab1, a human monoclonal antibody, was formulated at various concentrations, and included samples that contained <NUM> of either N-acetyl-arginine (NAR), arginine, N-acetyl-serine-arginine and N-acetyl-proline-arginine, or N-acetyl-arginine-NH<NUM> (this structure is shown as formula <NUM>) The results are shown in Table <NUM> and in <FIG>.

<FIG> is a line graph representation of the results shown in Table <NUM>, plotted against the results of a previous experiment using Ab1 formulated into <NUM> acetate/<NUM> N-acetyl-arginine/<NUM> Arg HCl, pH <NUM> (plot labeled "pH5. <NUM>"); <NUM> phosphate/<NUM> N-acetyl-arginine/<NUM> Arg HCl, pH <NUM> (plot labeled "pH6. <NUM>"); or <NUM> phosphate/<NUM> N-acetyl-arginine/<NUM> Arg HCl, pH <NUM> (plot labeled "pH6.

As shown in Table <NUM> and <FIG>, N-acetyl-proline-arginine dipeptide-containing sample ("PR") was observed to have a viscosity of <NUM> cP at <NUM>/ml of Ab1, and N-acetyl-serine-arginine dipeptide-containing sample ("SR") was observed to have a viscosity of <NUM> cP at <NUM>/ml of Ab1. The N-acetyl-arginine-containing sample ("NAR") had a viscosity of <NUM> cP at <NUM>/ml of Ab1, while the arginine-containing sample ("Arg") had a viscosity of <NUM> cP at <NUM>/ml of Ab1. In the case of the sample containing N-acetyl-arginine-NH<NUM> ("xRx"), viscosity was <NUM> cP at <NUM>/ml of Ab1, while the control containing no additional excipients (buffer and polysorbate <NUM>; "A52 PS80"), viscosity was observed to be <NUM> cP at <NUM>/ml of Ab1. Since <NUM> cP = <NUM> mPa·s, values referred to in cP equal the same values in mPa·s.

The objective of this experiment was to extend the observations shown in Example <NUM> by assaying additional therapeutic proteins and additional Arg-containing dipeptides.

Using the methods described in Example <NUM> and sample preparation as described in Example <NUM>, two additional therapeutic polypeptides, human antibodies Ab2 and Ab3, were tested at <NUM>/ml (+/-<NUM>%), <NUM>/ml (+/-<NUM>%), <NUM>/ml (+/-<NUM>%), and <NUM>/ml (+/- <NUM>%) (+/-<NUM>. The plan of the experiment is shown in Table <NUM>.

Viscosities of the three therapeutic polypeptides (human Abs: Ab1, Ab2, and Ab3) at the indicated concentrations (see Tables <NUM>-<NUM>) were measured in formulations containing the concentrations of the five excipients listed in Table <NUM>, as well as the absence of an excipient. Arginine HCl and glutamate-arginine were used as reference formulations. The results are presented in Tables <NUM> (Ab1), <NUM> (Ab2), and Ab3; as well as in the line graphs shown <FIG> (Ab1), <NUM> (Ab2), and <NUM> (Ab3). <FIG> and <FIG> include closer views (<FIG> and <FIG>) of the line graphs, as indicated by the boxes shown in <FIG> and <FIG>.

As shown for Ab1 (referring to <FIG> and the inset of <FIG> shown in <FIG>, the figure showing graphs of the data from Table <NUM>), without any excipients, viscosity greatly increases as the concentration of Ab1 exceeds <NUM>/mL (small filled circles in <FIG>). At <NUM>/mL, the viscosity is about <NUM> cP, but at about <NUM>/mL, the viscosity is almost <NUM> cP. The addition of Arg as Arg-HCl, N-acetyl-Pro-Arg, N-acetyl-Ser-Arg, and Glu-Arg decrease the viscosity of all tested samples at about <NUM>/mL Ab1 concentration, even at the highest Ab concentration of <NUM>/mL. However, Glu-Arg is less able to reduce the viscosity of the highest Ab1 concentration samples, having a viscosity of <NUM> cP at about <NUM>/mL of Ab1; interestingly, however, when the observed Ab1 concentration is decreased by about <NUM>/mL, the Glu-Arg dipeptide reduces the viscosity remarkably to about <NUM> cP (grey filled box in <FIG>). Surprisingly, N-acetyl-Pro-Arg-NH<NUM>, which contains an amide cap that reduces the overall charge on the molecule, also appeared to decrease the viscosity of the Ab1 samples, even at high concentrations of Ab1, such as about <NUM>/mL (empty triangles in <FIG>).

Likewise, as shown for Ab2, now referring to <FIG> and the inset of <FIG> as shown in <FIG> (the figure showing graphs of the data from Table <NUM>), without any excipients, viscosity greatly increases as the concentration of Ab2 exceeds <NUM>/mL (small filled circles in <FIG>). At <NUM>/mL, the viscosity is about <NUM> cP, but at about <NUM>/mL, the viscosity is about <NUM> cP. When formulated with the different excipients, viscosity was reduced for all samples, although less uniformly than was observed for Ab1. All of the N-acetylated dipeptides, including the amino capped dipeptide N-acetyl-Pro-Arg-NH<NUM>, decreased the viscosities of all samples having an antibody concentration greater than about <NUM>/mL.

A third Ab, Ab3 was also tested. Referring to <FIG>, which shows a line graph of the data shown in Table <NUM>, this Ab does not show as high viscosity as Abs <NUM> and <NUM>, with a viscosity of about <NUM> cP without any excipients. When viscosity was measured in these samples containing the indicated dipeptides and arginine, viscosity was reduced, but not as markedly as when the viscosity was greater. Even though the effect of the tested excipients is less notable, achieving a viscosity of, for example, about <NUM> cPs or less, can permit delivery by automated injection devices.

Claim 1:
A liquid pharmaceutical composition comprising a therapeutic polypeptide, a buffer, and at least one N-acetyl-dipeptide, wherein the N-acetyl-dipeptide is N-acetyl-serine-arginine, N-acetyl-proline-arginine, or N-acetyl-proline-arginine-NH<NUM>, optionally wherein the therapeutic polypeptide is an antibody or an antigen-binding fragment thereof.