Use of low molecular weight polyvinylpyrrolidone (PVP) to reduce the viscosity of highly concentrated protein formulations

Abstract

Disclosed herein are pharmaceutical compositions (formulations) in which low molecular weight polyvinylpyrrolidone (povidone) is used to reduce viscosity for high-concentration therapeutic protein (≧70 mg / mL) formulations. Addition of arginine, e.g., arginine monohydrochloride or N-acetylarginine, can further reduce viscosity. Suitable therapeutic proteins include antibodies, e.g., monoclonal antibodies, as well as derivatives, fragments, and analogs thereof.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 837,647, filed April 23, 2019, which is incorporated herein by reference.

[0002] The subject matter presented herein relates to the field of pharmaceutical formulations. In particular, the subject matter presented herein relates to high concentration therapeutic protein formulations and compositions and methods for reducing their viscosity using polyvinylpyrrolidone (PVP). [Background technology]

[0003] Pharmaceutically active proteins, such as antibodies, are often formulated in liquid solutions, particularly for injections. For example, for products that require administration via the subcutaneous route, such as those used for self-administration, formulations with delivery volumes exceeding 1-2 milliliters are not well tolerated. In such cases, highly concentrated protein formulations can accommodate the smaller desired volumes. The high dose and small volume requirements of such administration mean that protein therapeutics can reach concentrations of 100 mg / mL or much higher.

[0004] Highly concentrated protein formulations can pose many challenges to the manufacturability and administration of protein therapeutics. One challenge posed by some highly concentrated protein formulations is increased viscosity. Highly viscous formulations are difficult to handle during manufacturing, including the bulk and filling stages. Highly viscous formulations are also difficult to draw into a syringe and inject, making administration to patients difficult and uncomfortable. Summary of the Invention [Problem to be solved by the invention]

[0005] There is a need in the pharmaceutical industry to identify compounds useful for reducing the viscosity of highly concentrated protein formulations, to develop methods for reducing the viscosity of such formulations, and to provide pharmaceutical formulations with reduced viscosity. [Means for solving the problem]

[0006] In a first aspect, disclosed herein is a composition comprising a concentration of a therapeutic protein and polyvinylpyrrolidone (PVP), wherein the viscosity of the composition comprising PVP is lower than a composition comprising the same concentration of therapeutic protein but in the absence of PVP.

[0007] In a second aspect, disclosed herein is a composition comprising a concentration of a therapeutic protein and PVP, wherein the viscosity of the composition is 80 cP or less. The viscosity can be, for example, 70 cP, 40 cP, or 20 cP.

[0008] In both the first and second embodiments, the viscosity of the composition may be read at 25°C using, for example, an AR-G2 cone-and-plate rheometer manufactured by TA Instruments of New Castle, Delaware (USA) and reported at a shear rate of 1,000 / s. The therapeutic protein concentration may be greater than 70 mg / mL, such as from about 140 mg / mL to about 250 mg / mL, including 145 mg / mL, 160 mg / mL, 198 mg / mL, 200 mg / mL, 238 mg / mL, and 249 mg / mL. PVP may be present at concentrations of from about 0.3% to about 10%, such as about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10%, and increments therebetween. The stability of the therapeutic protein is approximately the same as that of a control lacking PVP; stability can be assessed by the presence of at least one selected from the group consisting of high molecular weight species, low molecular weight species, dimers, and oligomers of the therapeutic protein. In some embodiments, the therapeutic protein comprises at least one complementarity-determining region (CDR) and can be an antibody, such as a monoclonal antibody (mAb). Furthermore, the therapeutic protein comprising at least one CDR can be an antigen-binding fragment of an antibody or a derivative of an antibody. The antigen-binding fragment can be one selected from the group consisting of a Fab' fragment, an F'(ab)2 fragment, and an Fv fragment. In the case of an antibody derivative, the derivative can be selected from the group consisting of a humanized antibody, a chimeric antibody, a multispecific antibody, a maxibody, a BiTE® molecule, a single-chain antibody, a diabody, and a peptibody. PVP has a K value of 12 to 17, such as 12 or 17. The PVP may have a weight-average molecular weight of 11,000 Da or less, such as about 2,000 Da to about 25,000 Da, or about 2,000 Da to about 3,000 Da. The composition may be formulated for delivery to a patient. The formulation may have a pH of about 4.0 to about 8.0, such as about 4.6 to about 5.4. Additionally, the composition may contain arginine, such as N-acetylarginine (e.g., at a concentration of 10 mM), or a salt of arginine, such as arginine monohydrochloride (Arg-HCl), arginine glutamate, or arginine acetate. In the case of Arg-HCl, the Arg-HCl may be present at about 67 mM.In some embodiments where the composition includes Arg-HCL, PVP may be present at about 1%.

[0009] In sub-aspects of this first and second aspect, disclosed herein is a method of preparing a lyophilized powder comprising lyophilizing a composition of the first or second aspect.

[0010] In a third aspect, disclosed herein is a method for reducing the viscosity of a pharmaceutical formulation containing a therapeutic protein, the method comprising combining the therapeutic protein with a viscosity-reducing concentration of PVP. The viscosity of the composition produced by the method of this third aspect is 80 cP or less. The viscosity can be, for example, 70 cP, 40 cP, or 20 cP.

[0011] In this third aspect, the composition is made by the method of this aspect, and the viscosity may be read, for example, using an AR-G2 cone-and-plate rheometer manufactured by TA Instruments of New Castle, Delaware (USA) at 25°C and reported at a shear rate of 1,000 / s. The therapeutic protein concentration is greater than 70 mg / mL, e.g., from about 140 mg / mL to about 250 mg / mL, including about 145 mg / mL, 160 mg / mL, 198 mg / mL, 200 mg / mL, 238 mg / mL, and 249 mg / mL. PVP may be present at a concentration of about 0.3% to about 10%, e.g., about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10%, and increments therebetween. The stability of the therapeutic protein is approximately the same as that of a control lacking PVP; stability can be assessed by the presence of at least one selected from the group consisting of high molecular weight species, low molecular weight species, dimers, and oligomers of the therapeutic protein. In some embodiments, the therapeutic protein comprises at least one complementarity-determining region (CDR) and can be an antibody, such as a monoclonal antibody (mAb). Furthermore, the therapeutic protein comprising at least one CDR can be an antigen-binding fragment of an antibody or a derivative of an antibody. The antigen-binding fragment can be one selected from the group consisting of a Fab' fragment, an F'(ab)2 fragment, and an Fv fragment. In the case of an antibody derivative, the derivative can be selected from the group consisting of a humanized antibody, a chimeric antibody, a multispecific antibody, a maxibody, a BiTE® molecule, a single-chain antibody, a diabody, and a peptibody. PVP has a K value of 12 to 17, such as 12 or 17. The PVP may have a weight-average molecular weight of 11,000 Da or less, such as about 2,000 Da to about 25,000 Da, or about 2,000 Da to about 3,000 Da. The composition may be formulated for delivery to a patient. The formulation may have a pH of about 4.0 to about 8.0, such as about 4.6 to about 5.4. Additionally, the composition may contain arginine, such as N-acetylarginine (e.g., at a concentration of 10 mM), or a salt of arginine, such as arginine monohydrochloride (Arg-HCl), arginine glutamate, or arginine acetate. In the case of Arg-HCl, the Arg-HCl may be present at about 67 mM.In some embodiments where the composition includes Arg-HCL, PVP may be present at about 1%.

[0012] In a fourth aspect, disclosed herein is a lyophilized powder comprising a therapeutic protein and PVP, wherein the PVP is present in a weight:weight concentration effective to reduce viscosity after reconstitution with a diluent. In related sub-aspects, the PVP is present in a concentration of about 100 μg / mg of therapeutic protein to about 1 mg / mg of therapeutic protein. For example, the PVP is present in a concentration of about 200 μg / mg to about 500 μg / mg of therapeutic protein to about 1 mg / mg of therapeutic protein prior to reconstitution with a diluent.

[0013] When the lyophilized powder is reconstituted with the diluent of this fourth aspect, the viscosity of the composition is 80 cP or less. The viscosity may be, for example, 70 cP, 40 cP, or 20 cP.

[0014] In this fourth aspect, for compositions where the lyophilized powder is reconstituted with a diluent, the viscosity may be read, for example, using an AR-G2 cone-and-plate rheometer manufactured by TA Instruments of New Castle, Delaware (USA) at 25°C and reported at a shear rate of 1,000 / s. The therapeutic protein concentration may be greater than 70 mg / mL, such as from about 140 mg / mL to about 250 mg / mL, including 145 mg / mL, 160 mg / mL, 198 mg / mL, 200 mg / mL, 238 mg / mL, and 249 mg / mL. PVP may be present at concentrations from about 0.3% to about 10%, such as about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10%, and increments therebetween. The stability of the therapeutic protein is approximately the same as that of a control lacking PVP; stability can be assessed by the presence of at least one selected from the group consisting of high molecular weight species, low molecular weight species, dimers, and oligomers of the therapeutic protein. In some embodiments, the therapeutic protein comprises at least one complementarity-determining region (CDR) and can be an antibody, such as a monoclonal antibody (mAb). Furthermore, the therapeutic protein comprising at least one CDR can be an antigen-binding fragment of an antibody or a derivative of an antibody. The antigen-binding fragment can be one selected from the group consisting of a Fab' fragment, an F'(ab)2 fragment, and an Fv fragment. In the case of an antibody derivative, the derivative can be selected from the group consisting of a humanized antibody, a chimeric antibody, a multispecific antibody, a maxibody, a BiTE® molecule, a single-chain antibody, a diabody, and a peptibody. PVP has a K value of 12 to 17, such as 12 or 17. The PVP may have a weight-average molecular weight of 11,000 Da or less, such as about 2,000 Da to about 25,000 Da, or about 2,000 Da to about 3,000 Da. The composition may be formulated for delivery to a patient. The formulation may have a pH of about 4.0 to about 8.0, such as about 4.6 to about 5.4. Additionally, the composition may contain arginine, such as N-acetylarginine (e.g., at a concentration of 10 mM), or a salt of arginine, such as arginine monohydrochloride (Arg-HCl), arginine glutamate, or arginine acetate. In the case of Arg-HCl, the Arg-HCl may be present at about 67 mM.In some embodiments where the composition includes Arg-HCL, PVP may be present at about 1%.

[0015] In another aspect, provided herein is a method of reconstituting the lyophilized powder of the fourth aspect, comprising the step of adding a sterile aqueous diluent. [Brief explanation of the drawings]

[0016] [Figure 1] 1 is a graph showing the viscosity (units: cP (centipoise)) of mAb1, an IgG2 monoclonal antibody (mAb), at a concentration of 200 mg / mL in the presence of polyvinylpyrrolidone (PVP) K12. [Figure 2] 1 is a graph showing the effect of PVP K12 on the viscosity of a solution of mAb2, a therapeutic IgG1 mAb, at a concentration of 198 mg / mL. [Figure 3] 1 is a graph showing the effect of PVP K12 on the viscosity of a solution of mAb3, a therapeutic IgG1 mAb, at a concentration of 238 mg / mL. [Figure 4] 1 is a graph showing the effect of PVP K12 on the viscosity of a solution of mAb4, a therapeutic IgG1 mAb, at a concentration of 249 mg / mL. [Figure 5] 1 is a graph showing the effect of PVP K12 concentration on the viscosity of mAb1 at a concentration of 145 mg / mL. [Figure 6] 1 is a graph comparing the viscosity effect of PVP with two different molecular weights on a solution with a high concentration (160 mg / mL) of mAb1. [Figure 7] 1 is a graph showing the appearance of high molecular weight (HMW) species, low molecular weight (LMW) species, and oligomers and dimers of high concentrations of mAbs (mAbs 1, 3, 4, and 5) in the presence of PVP K12. DETAILED DESCRIPTION OF THE INVENTION

[0017] Highly concentrated protein formulations often face challenges due to increased viscosity. Highly viscous formulations are difficult to handle during manufacturing, including bulk and filling stages. Highly viscous formulations are also difficult to draw into a syringe and inject, often requiring the use of smaller gauge (larger bore) needles, which can be uncomfortable for patients (e.g., uncomfortable or even painful).

[0018] Disclosed herein are compositions and methods that take advantage of a surprising observation: the addition of low molecular weight polyvinylpyrrolidone (PVP) can reduce the viscosity of viscous therapeutic formulations. Commercially available PVPs are not sold as viscosity-reducing agents; in fact, some are sold as thickening agents (e.g., PVP K90; (2014)). PVPs are sold under various trademarks, including Kollidon® (BASF), a registered trademark for a range of pharmaceutical-grade PVPs.

[0019] For example, PVP K12 and PVP K17 are low molecular weight grades that are non-toxic and acceptable for use in parenteral formulations. Both PVP K12 and PVP K17 are commercially available as lyophilisers to stabilize fine molecular structures in lyophilisates and in injectable solutions; and as complexing and dissolution enhancers that form hydrogen bonds with compounds with complementary structures to improve dissolution (2014).

[0020] Disclosed herein are methods and compositions that take advantage of the surprising results that can reduce the viscosity of highly concentrated therapeutic protein compositions, such as compositions containing antibodies (e.g., monoclonal antibodies (mAbs) and their antigen-binding fragments, and derivatives and analogs thereof). In some cases, PVP is combined with arginine (e.g., a salt of arginine, e.g., arginine hydrochloride, Arg-HCl) to reduce viscosity even further, a surprising result that suggests that PVP and arginine act in a complementary manner.

[0021] Based on the results described in the Examples, PVP K12 is a preferred PVP (but not the only useful PVP) for reducing the viscosity of therapeutic proteins at high concentrations. For example, PVP K12 can be used at concentrations of 5% or less; PVP K12 can be used in combination with other excipients. Interestingly, a synergistic effect on viscosity reduction was observed when PVP K12 and arginine HCl were combined, indicating a low intrinsic viscosity and a low contribution to solution osmolality at the concentrations tested. PVP K12 does not appear to promote precipitation of therapeutic proteins or induce any significant adverse effects on protein stability at the low concentrations demonstrated to significantly reduce viscosity.

[0022] Composition ingredients and methods The following sections discuss PVP (and Arg-HCl), as well as suitable therapeutic proteins, viscosity, formulation preparation, pharmaceutical compositions, storage, and kits. Further definitions can be found in the Examples below.

[0023] Polyvinylpyrrolidone (PVP) Polyvinylpyrrolidone (PVP), also known as povidone, is a synthetic polymer vehicle often used to dispense and suspend drugs. PVP has many uses, including as a binder for tablets and capsules, a film-forming agent for eye drops, to aid in flavoring liquids and chewable tablets, and as an adhesive for transdermal systems.

[0024] Polyvinylpyrrolidone has the formula (C6H9NO) n means a molecule having the formula (1): [ka] It has the following structure.

[0025] PVP is also known as povidone, polypovidone, polyvidonum, poly(N-vinyl-2-pyrrolidinone), poly(N-vinylbutyrolactam), poly(1-vinyl-2-pyrrolidinone), 1-vinyl-2-pyrrolidinone homopolymer, and poly[1-(2-oxo-1-pyrrolidinyl)ethylene]. PVP is a highly polar, amphoteric, water-soluble polymer (polyamide). Purified PVP appears as a white to slightly off-white powder. PVP is often described using the k-value (Fikentscher K-value), which refers to the K-value viscosity of PVP. A higher K-value indicates a higher K-value viscosity. Commercially available PVP is available in various viscosity grades according to its K-value; e.g., PVP K15, K30, K60, and K90; see also Table 1.

[0026] The Fikentscher value of viscosity characteristics, K value, represents a viscosity index related to molecular weight, and is calculated using the relative viscosity measured by a capillary viscometer at 25°C according to the Fikentscher equation (2): K=(1.5logη rel -1) / (0.15+0.003c)+(300clogη rel +(c+1.5clogη rel ) 2 ) 1 / 2 / (0.15c+0.003c 2 )(2) (In the formula, η rel : Relative viscosity of PVP aqueous solution to water c: The PVP content in the PVP aqueous solution (weight / weight%).

[0027] Given that PVP is a polymer, its molecular weight can be determined via at least three different methods {Buehler, 2005 #12}: 1. The individual weights of the molecules are determined, e.g., by light scattering, M w The weight average molecular weight is indicated as (Table 1). 2.M nThe number average molecular weight is expressed as the number average molecular weight determined by methods such as osmometry, which measure the number of molecules. This number is very rarely determined or used for PVP. 3.M v The viscosity average molecular weight is expressed as σ and is determined by measuring the viscosity. This value can be calculated directly from the relative viscosity, intrinsic viscosity, or K value (Table 1).

[0028] A polymer consists of molecules with a range of molecular weights, in the ideal case with a Gaussian distribution {Buehler, 2005 #12}.

[0029] Kollidon® is a pharmaceutical grade of PVP marketed by BASF Corporation, Florham Park, NJ. Table 1 shows molecular weight values ​​for Kollidon grades.

[0030] PVP can be found in the form of monomers, dimers and polymers, as well as mixtures thereof.

[0031] [Table 1]

[0032] Accordingly, provided herein are methods for stabilizing or reducing the viscosity of protein formulations (pharmaceutical formulations / compositions) by adding PVP, and in some cases, Arg-HCl, in amounts effective to reduce viscosity. Additionally provided are low-viscosity formulations of therapeutic proteins, including antibodies (such as monoclonal antibodies (mAbs) and their antigen-binding fragments), containing effective amounts or concentrations of PVP and, in some cases, Arg-HCl. Also contemplated are methods of screening one or more formulations, each containing different concentrations of PVP (with and without Arg-HCl), to identify a suitable or optimal concentration for reducing viscosity. Additionally provided are methods of preparing lyophilized powders from the low-viscosity solution formulations described herein and methods of reconstituting such lyophilized powders by adding a (sterile) diluent.

[0033] Thus, provided herein are pharmaceutical formulations containing a biologically active (therapeutic) polypeptide and a viscosity-reducing concentration of PVP or a combination of PVP and Arg-HCl. The viscosity reduction is at least about 5-90% relative to a control formulation (e.g., lacking PVP and / or Arg-HCl). For example, the viscosity reduction can range from about 10% to about 80%. In other cases, the viscosity reduction can be at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, or 85% or more.

[0034] The concentration and grade of PVP (with or without Arg-HCl) for viscosity reduction can be determined empirically by one of ordinary skill in the art. In some embodiments, the PVP can have a concentration of about 0.3% to 10%, such as about 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%, and any increment therebetween.

[0035] The weight average molecular weight (M) of PVP in Da (Dalton) units w ) is about 2,000 Da to 25,000 Da, for example, 2,000 Da, 2500 Da, 3,000 Da, 3500 Da, 4,000 Da, 4500 Da, 5,000 Da, 5500 Da, 6,000 Da, 6500 Da, 7,000 Da, 7500 Da, 8,000 Da, 8500 Da, 9,000 Da, 9500 Da, 10,000 Da, 10,5 The molecular weight of the PVP may be 1000 Da, 11,000 Da, 12,000 Da, 13,000 Da, 14,000 Da, 15,000 Da, 16,000 Da, 17,000 Da, 18,000 Da, 19,000 Da, 20,000 Da, 21,000 Da, 22,000 Da, 23,000 Da, 24,000 Da, 25,000 Da, and any increment therebetween. In some embodiments, the PVP has a weight average molecular weight of 11,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 25,000 Da or less. In some embodiments, the PVP has a weight average molecular weight of 20,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 15,000 Da or less. In some embodiments, the PVP has a weight average molecular weight of 11,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 10,000 Da or less. In some embodiments, the PVP has a weight average molecular weight of 11,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 10,000 Da or less. In still other embodiments, the PVP has a weight average molecular weight of 9,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 8,000 Da or less. In still other embodiments, the PVP has a weight average molecular weight of 7,000 Da or less. In another embodiment, the PVP has a weight average molecular weight of 6,000 Da or less. In another embodiment, the PVP has a weight average molecular weight of 5,000 Da or less. In another embodiment, the PVP has a weight average molecular weight of 6,000 Da or less. In another embodiment, the PVP has a weight average molecular weight of 3,000 Da or less. And in some other embodiments, the PVP has a weight average molecular weight of 2,000 Da or less.

[0036] M is expressed as the viscosity average value of the molecular weight calculated from the K value of PVP. v is, for example, about 2,600 Da to about 5,500 Da (K12) (M calculated from the nominal K value of 3,900 Da) v (including K17) and about 7,100 Da to about 11,000 Da (M calculated from the nominal K value of 9,300 Da) vand about 2,600 Da to 25,000 Da, such as about 11,000 Da to about 25,000 Da. For example, the average viscosity values ​​of molecular weights are: 2,600 Da, 2,700 Da, 2,800 Da, 2,900 Da, 3,000 Da, 3,100 Da, 3,200 Da, 3,300 Da, 3,400 Da, 3,500 Da, 3,600 Da, 3,700 Da, 3,800 Da, 3,900 Da, 4,000 Da, 4,100 Da, 4,200 Da, 4,300 Da, 4,400 Da, 4,500 Da, 4,600 Da, 4,700 Da, 4,800 Da, 4,900 Da, 5,000 Da, 5,100 Da, 5,200 Da, 5,300 Da, 5,400 Da. , 5,500 Da, 6,000 Da, 7,000 Da, 7,100 Da, 7,500 Da, 8,000 Da, 8,500 Da, 9,000 Da, 9,500 Da, 10,000 Da, 10,500 Da, 11,000 Da, 12,000 Da, 13,000 Da, 14,000 Da, 15,000 Da, 16,000 Da, 17,000 Da, 18,000 Da, 19,000 Da, 20,000 Da, 21,000 Da, 22,000 Da, 23,000 Da, 24,000 Da, 25,000 Da and any increment therebetween. In some embodiments, the PVP has a weight average molecular weight of 11,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 25,000 Da or less. In some embodiments, the PVP has a weight average molecular weight of 20,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 15,000 Da or less. In some embodiments, the PVP has a weight average molecular weight of 11,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 10,000 Da or less. In still other embodiments, the PVP has a weight average molecular weight of 9,000 Da or less. In other embodiments, the PVP has a weight average molecular weight of 8,000 Da or less. In still other embodiments, the PVP has a weight average molecular weight of 7,000 Da or less. In another embodiment, the PVP has a weight average molecular weight of 6,000 Da or less. In another embodiment, the PVP has a weight average molecular weight of 5,000 Da or less. In another embodiment, the PVP has a weight average molecular weight of 6,000 Da or less.In another embodiment, the PVP has a weight average molecular weight of 3,000 Da or less, and in some other embodiments, the PVP has a weight average molecular weight of 2,600 Da or less.

[0037] In some embodiments, arginine is present. In some embodiments, arginine is present as a salt of arginine. In some embodiments, the arginine salt is Arg-HCl. In such embodiments, the concentration of Arg-HCl is, for example, 0.1 mM, 0.25 mM, 0.5 mM, 0.75 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 12.5 mM, 15 mM, 17.5 mM, 20 mM, 22.5 mM, 25 mM, 27.5 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, The concentration of Arg-HCl can be varied from about 0.1 mM to about 100 mM, including 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, and 100 mM, and increments therebetween; the concentration of Arg-HCl can further be 110 mM, 120 mM, 125 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 175 mM, 180 mM, 190 mM, and 200 mM; and increments therebetween. The arginine salt may also be Arg acetate or Arg glutamate and is present at a concentration of about 25 mM to about 150 mM, for example, about 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM, 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, or about 150 mM. N-acetylarginine, with or without a salt of arginine, may further be present in concentrations of about 25 mM to about 230 mM, for example, about 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM, , 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, 150 mM, 155 mM, 160 mM, 165 mM, 170 mM, 175 mM, 180 mM, 185 mM, 190 mM, 195 mM, 200 mM, 205 mM, 210 mM, 215 mM, 220 mM, 225 mM or about 230 mM and increments therebetween.

[0038] Therapeutic PolypeptidesTypical protein concentrations in the formulations can range from about 70 mg / mL to about 300 mg / mL, about 120 mg / mL to about 270 mg / mL, about 140 mg / mL to about 255 mg / mL, about 140 mg / mL to about 240 mg / mL, or about 140 mg / mL to about 220 mg / mL, or about 190 mg / mL to about 210 mg / mL. The protein concentration will depend on the end use of the pharmaceutical formulation and can be readily determined by one of skill in the art. Particularly contemplated protein concentrations are at least about 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, 100 mg / mL, 105 mg / mL, 110 mg / mL, 115 mg / mL, 120 mg / mL, 125 mg / mL, 130 mg / mL, 135 mg / mL, 140 mg / mL, 145 mg / mL, 150 mg / mL, 155 mg / mL, 160 mg / mL, 165 mg / mL, 170 mg / mL, 175 mg / mL, 180 mg / mL, 185 mg / mL, 185 mg / mL, 186 mg / mL, 187 mg / mL, 188 mg / mL, 189 mg / mL, 190 mg / mL, 191 mg / mL, 192 mg / mL, 193 mg / mL, 194 mg / mL, 195 mg / mL, 196 mg / mL, 197 mg / mL, 198 mg / mL, 199 mg / mL, 200 mg / mL, 201 mg / mL, 202 mg / mL, 203 mg / mL, 204 mg / mL, 205 mg / mL, 206 mg / mL, 207 mg / mL, 208 mg / mL, 209 mg / mL, 210 mg / mL, 211 mg / mL, 212 mg / mL, 213 mg / mL, 214 mg / mL, 215 mg / mL, 216 mg / mL, 217 mg / mL, 218 mg / mL, 219 mg / mL, 220 mg / mL, 225 mg / mL, 225 mg / mL, 230 mg / mL, mg / mL, 175mg / mL, 180mg / mL, 185mg / mL, 190mg / mL, 191mg / mL, 192mg / mL, 193mg / mL, 194mg / mL, 195mg / mL, 196mg / mL, 197m g / mL, 198mg / mL, 199mg / mL, 200mg / mL, 201mg / mL, 202mg / mL, 203mg / mL, 204mg / mL, 205mg / mL, 206mg / mL, 207mg / mL, 208mg / mL, 209mg / mL, 210mg / mL, 211mg / mL, 212mg / mL, 213mg / mL, 214mg / mL, 215mg / mL, 216mg / mL, 217mg / mL, 218mg / mL, 219mg / mL mL, 220mg / mL, 221mg / mL, 222mg / mL, 223mg / mL, 224mg / mL, 225mg / mL, 226mg / mL, 227mg / mL, 228mg / mL, 229mg / mL, 230mg / m L, 231mg / mL, 232mg / mL, 233mg / mL, 234mg / mL, 235mg / mL, 236mg / mL, 237mg / mL, 238mg / mL, 239mg / mL, 240mg / mL, 241mg / mL , 242mg / mL, 243mg / mL, 244mg / mL, 245mg / mL, 246mg / mL, 247mg / mL, 248mg / mL, 249mg / mL, 250mg / mL, 251mg / mL, 252mg / mL,253mg / mL, 254mg / mL, 255mg / mL, 256mg / mL, 257mg / mL, 258mg / mL, 259mg / mL, 260mg / mL, 261mg / mL, 262mg / mL, 263mg / mL, 264mg / mL, 2 65mg / mL, 266mg / mL, 267mg / mL, 268mg / mL, 269mg / mL, 270mg / mL, 271mg / mL, 272mg / mL, 273mg / mL, 274mg / mL, 275mg / mL, 276mg / mL, 27 7mg / mL, 278mg / mL, 279mg / mL, 280mg / mL, 281mg / mL, 282mg / mL, 283mg / mL, 284mg / mL, 285mg / mL, 286mg / mL, 287mg / mL, 288mg / mL, 289 mg / mL, 290 mg / mL, 291 mg / mL, 292 mg / mL, 293 mg / mL, 294 mg / mL, 295 mg / mL, 296 mg / mL, 297 mg / mL, 298 mg / mL, 299 mg / mL and 300 mg / mL. ,

[0039] Viscosity and other properties of PVP-containing formulations In one aspect, the pharmaceutical formulations disclosed herein (including PVP, and with or without Arg-HCl) have a viscosity level of less than about 80 cP (centipoise) when measured at room temperature (i.e., 25° C.). In certain embodiments, the pharmaceutical formulations have a viscosity level of less than about 80 cP to less than about 1 cP, e.g., 80 cP, 70 cP, about 60 cP, about 50 cP, about 40 cP, about 30 cP, about 25 cP, about 20 cP, about 18 cP, about 15 cP, about 12 cP, about 10 cP; about 8 cP, about 6 cP, about 4 cP; about 2 cP; or about 1 cP.

[0040] In one aspect, a pharmaceutical formulation is stable as measured by at least one stability assay, e.g., an assay that tests a biophysical or biochemical property of a therapeutic protein (e.g., an antibody) over time. A "stable pharmaceutical formulation" or "stable formulation" refers to a pharmaceutical formulation containing a therapeutic protein that exhibits a limited increase in aggregation and / or a reduced loss of biological activity of 5% to 10% or less when stored at about -30°C (or lower) to about 5°C to about 40°C for at least one month, or two months, or three months, or six months, or one year, or two years, or five years, or more, when compared to a control formulation sample. Formulation stability can be determined using any number of standard assays, including size exclusion HPLC (SEC-HPLC), cation exchange HPLC (CEX-HPLC), subvisible detection by light obscuration technology ("HIAC"), and / or visual inspection. Typically, higher storage temperatures result in shorter shelf-life formulations.

[0041] Pharmaceutical formulation stability can also be evaluated using visual evaluation. Visual evaluation is a qualitative method used to describe the visible physical properties of a sample. Samples are inspected against a black and / or white background in an inspection booth, depending on the properties being evaluated (e.g., color, clarity, presence of particles or foreign matter). Samples are also inspected against an opalescent reference standard and a color reference standard. In the case of visual evaluation, a stable pharmaceutical formulation does not show significant changes in color, clarity, presence of particles or foreign matter compared to a control sample.

[0042] The formulation can have any pH that is appropriate for the therapeutic polypeptide to maintain activity and acceptable stability while also being suitable for administration to a patient. For example, the pH can be from about 4.0 to about 8.0, such as about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, and 8.0. In some cases, the pH range is from about 4.6 to about 5.4.

[0043] Pharmaceutical composition formulations and ingredients Pharmaceutical compositions suitable for administration to a patient can be prepared containing other ingredients in addition to PVP (and in some cases, arginine, such as Arg-HCl).

[0044] Acceptable pharmaceutical ingredients are preferably non-toxic to patients at the dosages and concentrations employed. Pharmaceutical compositions can include agents to alter, maintain, or protect, for example, the pH, osmolality, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption, or permeability of the composition.

[0045] In general, excipients can be classified based on the mechanism by which they stabilize proteins against various chemical and physical stresses. Some excipients mitigate the effects of specific stresses or modulate the specific susceptibility of specific polypeptides. Other excipients have a more general effect on the physical and covalent stability of proteins.

[0046] Common excipients for liquid and lyophilized protein formulations are listed in Table A (see also (Kamerzell et al 2011)).

[0047] [Table 2]

[0048] [Table 3]

[0049] Other excipients are known in the art (see, e.g., (Powell et al 1998)). One of skill in the art can determine what amount or range of excipients may be included in any particular formulation to obtain a biopharmaceutical composition of the invention that promotes the retention of stability of the biopharmaceutical. For example, the amount and type of salt to include in a biopharmaceutical composition of the invention can be selected based on the desired osmolality (i.e., isotonic, hypotonic, or hypertonic) of the final solution and the amount and osmolality of other components to be included in the formulation.

[0050] Preparation of Polypeptide Formulations The pharmaceutical formulations disclosed herein can be prepared by either of two processes designated Process 1 and Process 2. Process 1 comprises: a. Dialyzing or concentrating the therapeutic protein solution; b. Dialyzing or concentrating a solution of the selected excipients or providing a dry mix of the selected excipients; c. adding an excipient solution or dry excipient mixture at a selected pH to a protein solution to achieve a desired final excipient concentration, a desired final protein concentration, and a desired final pH; d. UF / DF The ultrafiltration dialysis process exchanges buffer and simultaneously concentrates proteins.

[0051] Process 2 is: a. dialyzing the therapeutic protein solution; b. Dialyzing a solution of the selected excipients or providing a dry mix of the selected excipients; c. adding an excipient solution or dry excipient mixture to the dialyzed protein solution at a selected pH and desired excipient concentration; and d. Concentrating the solution resulting from step c to a desired final protein concentration and a desired final pH.

[0052] In Process 1, the pH of the concentrated protein to achieve the desired final pH can range from about 4 to about 8. In Process 2, the pH of the concentrated protein solution to achieve the desired final pH can range from about 4 to about 8. When a particular excipient is reported in a formulation, for example, as a weight / volume %, one of skill in the art will recognize that equimolar concentrations of that excipient are also contemplated.

[0053] Formulations can be lyophilized for subsequent resuspension with an appropriate diluent; liquid formulations are often modified to incorporate cryoprotectants and bulking agents; acetate is replaced with glutamate or phosphate to reduce volatility.

[0054] Storage and Kits Once the pharmaceutical formulation has been formulated, it may be stored in a sterile vial as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored either in a ready-to-use form or in a form that is reconstituted prior to administration (e.g., lyophilized). In some cases, the therapeutic polypeptide formulation may be stored in a container such as a suitable storage bag (e.g., manufactured by Sartorius, Gottingen, DE) or a polycarbonate carboy. Once the pharmaceutical formulation has been formulated, it may also be stored in a ready-to-use form, as a solution or suspension in a glass vial (e.g., a 5 cc glass vial), or in a prefilled syringe (PFS; e.g., a 2.25 mL PFS).

[0055] In certain embodiments, kits for producing single-dose administration units are provided. In certain embodiments, the kits may contain both a first container with a dried protein and a second container with an aqueous formulation. In certain embodiments, kits are provided that include single-chamber and multi-chamber pre-filled syringes (e.g., liquid syringes and lyosyringes).

[0056] Embodiment Embodiment 1: A composition comprising a concentration of a therapeutic protein and polyvinylpyrrolidone (PVP), wherein the viscosity of the composition with PVP is lower than a composition with the same concentration of therapeutic protein without the PVP present.

[0057] Embodiment 2: A composition comprising a concentration of a therapeutic protein and PVP, wherein the viscosity of the composition is 80 cP or less.

[0058] Embodiment 3: The composition of embodiment 2, wherein the viscosity of the composition is 70 cP or less.

[0059] Embodiment 4: The composition of embodiment 2, wherein the viscosity of the composition is 40 cP or less.

[0060] Embodiment 5: The composition of embodiment 2, wherein the viscosity of the composition is 20 cP or less.

[0061] Embodiment 6: The composition of embodiment 1 or 2, wherein the viscosity of the composition is read at 25°C and reported at a shear rate of 1,000 / s.

[0062] Embodiment 7: The composition of embodiment 6, wherein the viscosity is measured using an AR-G2 cone and plate rheometer manufactured by TA Instruments, New Castle, Delaware (USA).

[0063] Embodiment 8: The composition of embodiment 1 or 2, wherein the concentration of the therapeutic protein is greater than 70 mg / mL.

[0064] Embodiment 9: The composition of embodiment 8, wherein the concentration of the therapeutic protein is greater than or equal to about 140 mg / mL to about 250 mg / mL.

[0065] Embodiment 10: The composition of embodiment 9, wherein the concentration of the therapeutic protein is selected from the group consisting of about 145 mg / mL, 160 mg / mL, 198 mg / mL, 200 mg / mL, 238 mg / mL, and 249 mg / mL.

[0066] Embodiment 11: The composition of embodiment 1 or 2, wherein the PVP is present in a concentration of about 0.3% to about 10%.

[0067] Embodiment 12: The composition of embodiment 11, wherein the PVP is present at a concentration selected from the group consisting of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10% and increments therebetween.

[0068] Embodiment 13: The composition of embodiment 1 or 2, wherein the stability of the therapeutic protein is about the same as compared to a control lacking PVP.

[0069] Embodiment 14: The composition of embodiment 13, wherein the stability is assessed by the presence of at least one selected from the group consisting of high molecular weight species, low molecular weight species, dimers, and oligomers.

[0070] Embodiment 15: The composition of embodiment 1 or 2, wherein the therapeutic protein comprises at least one complementarity determining region (CDR).

[0071] Embodiment 16: The composition of embodiment 15, wherein the therapeutic protein is an antibody.

[0072] Embodiment 17: The composition of embodiment 16, wherein the antibody is a monoclonal antibody (mAb).

[0073] Embodiment 18: The composition of embodiment 17, wherein the antibody is an antigen-binding fragment or derivative of an antibody.

[0074] Embodiment 19: The composition of embodiment 18, wherein the antigen-binding fragment is selected from the group consisting of a Fab' fragment, an F'(ab)2 fragment, and an Fv fragment.

[0075] Embodiment 20: The composition of embodiment 18, wherein the antibody derivative is selected from the group consisting of a humanized antibody, a chimeric antibody, a multispecific antibody, a maxibody, a BiTE® molecule, a single-chain antibody, a diabody, and a peptibody.

[0076] Embodiment 21: The composition of embodiment 1 or 2, wherein the PVP has a K value of 12 to 17, for example, 12 or 17.

[0077] Embodiment 22: The composition of embodiment 1 or 2, wherein the PVP has a weight average molecular weight of 11,000 Da or less.

[0078] Embodiment 23: The composition of embodiment 23, wherein the PVP has a weight average molecular weight of about 2,000 Da to about 25,000 Da.

[0079] Embodiment 24: The composition of embodiment 24, wherein the PVP has a weight average molecular weight of about 2,000 Da to about 3,000 Da.

[0080] Embodiment 25: The composition of embodiment 1 or 2, wherein the composition is formulated for delivery to a patient.

[0081] Embodiment 26: The composition of embodiment 1 or 2, having a pH of about 4.0 to about 8.0.

[0082] Embodiment 27: The composition of embodiment 26, having a pH of about 4.6 to about 5.4.

[0083] Embodiment 28: The composition of embodiment 1 or 2, further comprising arginine.

[0084] Embodiment 29: The composition of embodiment 28, wherein the arginine is N-acetylarginine.

[0085] Embodiment 30: The composition of embodiment 29, wherein N-acetylarginine is present at about 10 mM.

[0086] Embodiment 31: The composition of embodiment 28, wherein the arginine is a salt of arginine.

[0087] Embodiment 32: The composition of embodiment 31, wherein the arginine is arginine monohydrochloride (Arg-HCl), arginine glutamate, or arginine acetate.

[0088] Embodiment 33: The composition of embodiment 32, wherein Arg-HCl is present at about 67 mM.

[0089] Embodiment 34: The composition of embodiment 33, wherein PVP is present at about 1%.

[0090] Embodiment 35: A method of preparing a lyophilized powder, comprising lyophilizing the composition of embodiment 1 or 2.

[0091] Embodiment 36: A method of reducing the viscosity of a pharmaceutical formulation comprising a therapeutic protein, comprising associating the therapeutic protein with a viscosity-reducing concentration of PVP.

[0092] Embodiment 37: The method of embodiment 36, wherein the viscosity of the composition is 80 cP or less.

[0093] Embodiment 38: The method of embodiment 36, wherein the viscosity of the composition is 70 cP or less.

[0094] Embodiment 39: The method of embodiment 36, wherein the viscosity of the composition is 40 cP or less.

[0095] Embodiment 40: The method of embodiment 36, wherein the viscosity of the composition is 20 cP or less.

[0096] Embodiment 41: The method of embodiment 36, wherein the viscosity of the composition is read at 25°C and reported at a shear rate of 1,000 / s.

[0097] Embodiment 42: The method of embodiment 42, wherein the viscosity is measured using an AR-G2 cone-and-plate rheometer manufactured by TA Instruments, New Castle, Delaware (USA).

[0098] Embodiment 43: The method of embodiment 36, wherein the concentration of the therapeutic protein is greater than about 70 mg / mL.

[0099] Embodiment 44: The method of embodiment 43, wherein the concentration of the therapeutic protein is greater than or equal to about 140 mg / mL to about 250 mg / mL.

[0100] Embodiment 45: The method of embodiment 44, wherein the concentration of the therapeutic protein is selected from the group consisting of about 145 mg / mL, 160 mg / mL, 198 mg / mL, 200 mg / mL, 238 mg / mL, and 249 mg / mL.

[0101] Embodiment 46: The method of embodiment 36, wherein the PVP is present at a concentration of about 0.3% to about 10%.

[0102] Embodiment 47: The method of embodiment 46, wherein the PVP is present at a concentration selected from the group consisting of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10% and increments therebetween.

[0103] Embodiment 48: The method of embodiment 36, wherein the stability of the therapeutic protein is about the same as compared to a control lacking PVP.

[0104] Embodiment 49: The method of embodiment 48, wherein the stability is assessed by the presence of at least one selected from the group consisting of high molecular weight species, low molecular weight species, dimers, and oligomers.

[0105] Embodiment 50: The method of embodiment 36, wherein the therapeutic protein comprises at least one complementarity determining region (CDR).

[0106] Embodiment 51: The method of embodiment 50, wherein the therapeutic protein is an antibody.

[0107] Embodiment 52: The method of embodiment 51, wherein the antibody is a monoclonal antibody (mAb).

[0108] Embodiment 53: The method of embodiment 51, wherein the antibody is an antigen-binding fragment of an antibody or a derivative of an antibody.

[0109] Embodiment 54: The method of embodiment 53, wherein the antigen-binding fragment is selected from the group consisting of a Fab' fragment, an F'(ab)2 fragment, and an Fv fragment.

[0110] Embodiment 55: The method of embodiment 53, wherein the antibody derivative is selected from the group consisting of a humanized antibody, a chimeric antibody, a multispecific antibody, a maxibody, a BiTE® molecule, a single-chain antibody, a diabody, and a peptibody.

[0111] Embodiment 56: The method of embodiment 36, wherein the PVP has a K value of 12 to 17, for example 12 or 17.

[0112] Embodiment 57: The method of embodiment 36, wherein the PVP has a weight average molecular weight of 11,000 Da or less.

[0113] Embodiment 58: The method of embodiment 57, wherein the PVP has a weight average molecular weight of about 2,000 Da to about 25,000 Da.

[0114] Embodiment 59: The method of embodiment 58, wherein the PVP has a weight average molecular weight of about 2,000 Da to about 3,000 Da.

[0115] Embodiment 60: The method of embodiment 36, wherein the composition is formulated for delivery to a patient.

[0116] Embodiment 61: The method of embodiment 36, wherein the composition has a pH of about 4.0 to about 8.0 after reconstitution with a diluent.

[0117] Embodiment 62: The method of embodiment 61, wherein the composition has a pH of about 4.6 to about 5.4.

[0118] Embodiment 63: The method of embodiment 33, wherein the composition further comprises arginine.

[0119] Embodiment 64: The method of embodiment 64, wherein the arginine is N-acetylarginine.

[0120] Embodiment 65: The method of embodiment 65, wherein N-acetylarginine is present at about 10 mM.

[0121] Embodiment 66: The method of embodiment 63, wherein the arginine is a salt of arginine.

[0122] Embodiment 67: The method of embodiment 66, wherein the arginine is arginine monohydrochloride (Arg-HCl), arginine glutamate, or arginine acetate.

[0123] Embodiment 68: The method of embodiment 67, wherein Arg-HCl is present at about 67 mM.

[0124] Embodiment 69: The method of embodiment 68, wherein PVP is present at about 1%.

[0125] Embodiment 70: A lyophilized powder comprising a therapeutic protein and PVP, wherein the PVP is present in a weight:weight concentration effective to reduce viscosity after reconstitution with a diluent.

[0126] Embodiment 71: The lyophilized powder of embodiment 70, wherein the PVP is present in a concentration of about 100 μg / mg of therapeutic protein to about 1 mg / mg of therapeutic protein.

[0127] Embodiment 72: The lyophilized powder of embodiment 71, wherein the PVP is present in a concentration of from about 200 μg / mg to about 500 μg / mg of therapeutic protein to about 1 mg / mg of therapeutic protein prior to reconstitution with a diluent.

[0128] Embodiment 73: The lyophilized powder of embodiment 71, wherein the viscosity of the method is 80 cP or less after reconstitution with a diluent.

[0129] Embodiment 74: The lyophilized powder of embodiment 71, wherein the viscosity of the powder is 70 cP or less after reconstitution with a diluent.

[0130] Embodiment 75: The lyophilized powder of embodiment 71, wherein the viscosity of the powder is 40 cP or less after reconstitution with a diluent.

[0131] Embodiment 76: The lyophilized powder of embodiment 71, wherein the viscosity of the method is 20 cP or less after reconstitution with a diluent.

[0132] Embodiment 77: The lyophilized powder of embodiment 71, wherein the viscosity of the method is read at 25°C after reconstitution with a diluent and is reported at a shear rate of 1,000 / s.

[0133] Embodiment 78: The freeze-dried powder of embodiment 71, wherein the viscosity is measured using an AR-G2 cone-and-plate rheometer manufactured by TA Instruments, New Castle, Delaware (USA).

[0134] Embodiment 79: The lyophilized powder of embodiment 70, wherein the PVP is present in a concentration of about 0.3% to about 10% after reconstitution with a diluent.

[0135] Embodiment 80: The lyophilized powder of embodiment 79, wherein the PVP is present at a concentration selected from the group consisting of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10% after reconstitution with the diluent and increments therebetween.

[0136] Embodiment 81: The lyophilized powder of embodiment 70, wherein the therapeutic protein comprises at least one complementarity determining region (CDR).

[0137] Embodiment 82: The lyophilized powder of embodiment 81, wherein the therapeutic protein is an antibody.

[0138] Embodiment 83: The lyophilized powder of embodiment 82, wherein the antibody is a monoclonal antibody (mAb).

[0139] Embodiment 84: The lyophilized powder of embodiment 82, wherein the antibody is an antigen-binding fragment of an antibody or a derivative of an antibody.

[0140] Embodiment 85: The lyophilized powder of embodiment 84, wherein the antigen-binding fragment is selected from the group consisting of a Fab' fragment, an F'(ab)2 fragment, and an Fv fragment.

[0141] Embodiment 86: The lyophilized powder of embodiment 85, wherein the antibody derivative is selected from the group consisting of humanized antibodies, chimeric antibodies, multispecific antibodies, maxibodies, BiTE® molecules, single chain antibodies, diabodies and peptibodies.

[0142] Embodiment 87: The lyophilized powder of embodiment 70, wherein the PVP has a K value of 12 to 17, for example 12 or 17.

[0143] Embodiment 88: The lyophilized powder of embodiment 70, wherein the PVP has a weight average molecular weight of 11,000 Da or less.

[0144] Embodiment 89: The lyophilized powder of embodiment 88, wherein the PVP has a weight average molecular weight of about 2,000 Da to about 25,000 Da.

[0145] Embodiment 90: The lyophilized powder of embodiment 89, wherein the PVP has a weight average molecular weight of about 2,000 Da to about 3,000 Da.

[0146] Embodiment 91: The method comprises the lyophilized powder of embodiment 70, which is formulated for delivery to a patient after reconstitution with a diluent.

[0147] Embodiment 92: The lyophilized powder of embodiment 70, having a pH of about 4.0 to about 8.0 after reconstitution with a diluent.

[0148] Embodiment 93: The lyophilized powder of embodiment 92, having a pH of about 4.6 to about 5.4.

[0149] Embodiment 94: The lyophilized powder of embodiment 70, further comprising arginine.

[0150] Embodiment 95: The lyophilized powder of embodiment 94, wherein the arginine is N-acetylarginine.

[0151] Embodiment 96: The lyophilized powder of embodiment 95, wherein N-acetylarginine is present at about 10 mM.

[0152] Embodiment 97: The lyophilized powder of embodiment 94, wherein the arginine is a salt of arginine.

[0153] Embodiment 98: The lyophilized powder of embodiment 97, wherein the arginine is arginine monohydrochloride (Arg-HCl), arginine glutamate, or arginine acetate.

[0154] Embodiment 99: The lyophilized powder of embodiment 98, wherein arginine hydrochloride is present at about 67 mM.

[0155] Embodiment 100: The lyophilized powder of embodiment 99, wherein PVP is present at about 1%.

[0156] Embodiment 101: A method for reconstituting the lyophilized powder of embodiment 70, comprising adding a sterile aqueous diluent.

[0157] The following examples are offered by way of illustration only and are not intended to limit the disclosure or claims herein in any way. [Example]

[0158] Example 1 - PVP K12 as a viscosity-reducing excipient (with and without arginine-HCl) in a highly concentrated mAb solution (mAb1) To assess the effect of PVP K12 on the viscosity of a highly concentrated therapeutic mAb (IgG2), mAb1, mAb1 was dialyzed against 15 mM sodium acetate (pH 5.2). After dialysis, mAb1 was concentrated to 220 mg / mL using an Amicon® Ultra 10K molecular weight cutoff (MWCO) centrifugal filter (Millipore Sigma; Burlington, MA). Concentrated excipient stock solutions (PVP K12 (BASF Corp., headquartered in Ludwigshafen, Germany); and arginine HCl (Sigma-Aldrich; St. Louis, MO) were then spiked into this material at 10% by volume, diluting the mAb1 concentration to 200 mg / mL. The viscosity of each sample was measured at 25°C using an AR-G2 cone-plate rheometer (TA Instruments; New Castle, DE), and the data was reported at a shear rate of 1,000 / s. The data in Figure 1 show that the addition of 1% and 3% PVP K12 resulted in a substantial reduction in the viscosity of the mAb1 formulation. The amount of reduction for 3% PVP K12 is comparable to the level seen with the addition of 67 mM arginine HCl (Arg-HCl), which was used for comparison. The combination of K12 with 67 mM arginine HCl unexpectedly showed a further reduction in viscosity compared to formulations containing a single excipient.

[0159] Example 2 - PVP K12 as a viscosity-reducing excipient (with and without arginine-HCl) in a highly concentrated mAb solution (mAb2) Experiments were conducted to evaluate the effect of PVP K12 on the viscosity of a highly concentrated therapeutic mAb, mAb2 (IgG1). mAb2 was dialyzed against 10 mM sodium acetate (pH 5.2) containing 10 mM N-acetylarginine (NAR). After dialysis, mAb2 was concentrated to 220 mg / mL using an Amicon Ultra 10K MWCO centrifugal filter. Concentrated excipient stock solutions were then spiked into this material at 10% by volume, diluting the mAb2 concentration to 198 mg / mL. The viscosity of the samples was measured at 25°C using an AR-G2 cone-plate rheometer, and data are reported at a shear rate of 1,000 s. The data in Figure 2 show that the addition of 0.3%, 1%, and 3% PVP K12 (in the presence of 10 mM NAR) resulted in a reduction in the viscosity of the mAb2 formulation, with the 3% PVP formulation producing the lowest viscosity among the three PVP concentrations tested. The amount of reduction for 3% PVP K12 was comparable to the level seen with the addition of 67 mM arginine HCl, which was included for comparison. The combination of 1% PVP K12 and 67 mM arginine HCl unexpectedly showed a further reduction in viscosity compared to formulations containing a single excipient.

[0160] Example 3 - PVP K12 as a viscosity-reducing excipient (with and without arginine-HCl) in a highly concentrated mAb solution (mAb3) An experiment was conducted to evaluate the effect of PVP K12 on the viscosity of a highly concentrated therapeutic mAb, mAb3 (IgG1). mAb3 was dialyzed against 15 mM sodium acetate (pH 5.2). After dialysis, mAb3 was concentrated to 265 mg / mL using an Amicon Ultra 10K MWCO centrifugal filter. Concentrated excipient stock solution was then spiked into this material at 10% by volume, diluting the mAb3 concentration to 238 mg / mL. The viscosity of the samples was measured at 25°C using an AR-G2 cone-and-plate rheometer, and data were reported at a shear rate of 1,000 / s. The data in Figure 3 show that the addition of 0.3%, 1%, and 3% PVP K12 resulted in a decrease in the viscosity of the mAb3 formulation, with the 1% PVP K12 formulation producing the lowest viscosity among the three PVP concentrations tested. The combination of 1% PVP K12 and 67 mM arginine HCl unexpectedly showed a further reduction in viscosity compared to formulations containing a single excipient.

[0161] Example 4 - PVP K12 as a viscosity-reducing excipient (with and without arginine-HCl) in a highly concentrated mAb solution (mAb4) Experiments were conducted to evaluate the effect of PVP K12 on the viscosity of a therapeutic mAb, mAb4 (IgG1). mAb4 was dialyzed against 15 mM sodium acetate (pH 5.2). After dialysis, mAb3 was concentrated to 277 mg / mL using an Amicon Ultra 10K MWCO centrifugal filter. Concentrated excipient stocks were then spiked into this material at 10% by volume, diluting the mAb4 concentration to 249 mg / mL. The viscosity of the samples was measured at 25°C using an AR-G2 cone-and-plate rheometer, and data were reported at a shear rate of 1,000 / s. The data in Figure 4 show that the addition of 0.3%, 1%, and 3% PVP K12 resulted in a decrease in the viscosity of the mAb4 formulation, with the 1% PVP formulation producing the lowest viscosity among the three PVP concentrations tested. The amount of reduction for 1% PVP K12 was comparable to the level seen with the addition of 67 mM arginine HCl, which was included for comparison. The combination of 1% PVP K12 and 67 mM arginine HCl unexpectedly showed a further reduction in viscosity compared to formulations containing a single excipient.

[0162] Example 5 - Effect of different PVP K12 concentrations on a highly concentrated mAb solution (mAb1) Experiments were conducted to evaluate the effect of varying concentrations of PVP K12 on viscosity reduction of mAb1 formulations. mAb1 was dialyzed against 15 mM sodium acetate (pH 5.2) and concentrated to 181 mg / mL using an Amicon Ultra 10K MWCO centrifugal filter. A 50% wt / vol solution of PVP K12 was then spiked into the concentrated protein solution to produce PVP concentrations ranging up to 10%. The final mAb concentration was 145 mg / mL. The viscosity of the samples was measured at 25°C using an AR-G2 cone-plate rheometer, and data were reported at a shear rate of 1,000 / s. The data in Figure 5 show that as the PVP K12 concentration increased, viscosity reduction began to decrease when the PVP K12 concentration was ≥3%. The minimum viscosity was achieved between 5% and 10% PVP K12, with the viscosity increasing from 7.5% to 10% PVP K12.

[0163] Example 6 - Comparison of the effect on viscosity using various molecular weight PVPs in a highly concentrated mAb solution (mAb1) An experiment was conducted to compare the effect of various molecular weights of PVP on the viscosity of mAb1 formulations. mAb1 was dialyzed against 15 mM sodium acetate (pH 5.2). After dialysis, mAb1 was concentrated to 178 mg / mL using an Amicon Ultra 10K MWCO centrifugal filter. Concentrated excipient stock solutions of PVP K12 (MW: 2,000-3,000 Da) and PVP K17 (MW: 7,000-11,000 Da) (all from BASF Corp.) were then spiked into this material at 10% by volume, diluting the mAb1 concentration to 160 mg / mL. The viscosity of the samples was measured at 25 °C using an AR-G2 cone-plate rheometer, and data are reported at a shear rate of 1,000 s. The data in Figure 6 show that PVP K12, which has a lower average molecular weight compared to PVP K17, was a more effective viscosity-reducing excipient than PVP K17 at comparable concentrations.

[0164] Example 7 – Stability of mAb in formulations containing PVP K12 Experiments were conducted to evaluate the effect of 2% PVP K12 on the stability of several mAbs. mAbs (including mAb5 and IgG2) were dialyzed against 15 mM sodium acetate (pH 5.2). After dialysis, PVP K12 was spiked to a final concentration of 2%. The mAb concentration was adjusted to 100 mg / mL, and the samples were incubated at 40°C for 2 weeks before analysis by size-exclusion high-performance liquid chromatography (SE-HPLC). Figure 7 shows a graph of the SE-HPLC area percentages of several degradants for a 2% PVP K12 mAb compared to a water-spiked control. These results indicate that 2% PVP K12 did not significantly affect the stability of the mAbs. This observation suggests that viscosity-reducing effects can be applied without any significant increase in aggregation or clipping.

[0165] definition "Viscosity" is the resistance of a fluid to flow and can be measured in centipoise (cP) or millipascal-seconds (mPa-s), where 1 cP = 1 mPa-s, at a given shear rate. Viscosity can be measured using a viscometer, such as a Brookfield Engineering Dial Reading Viscometer Model LVT (AMETEK Brookfield, Middleboro, MA) and an AR-G2 Cone-and-Plate Rheometer (TA Instruments; New Castle, DE). In some cases, viscosity is measured at 25°C and reported at a shear rate of 1,000 / s. Viscosity can also be measured using any other method and any other unit known in the art (e.g., absolute viscosity, kinematic viscosity, or dynamic viscosity), with the understanding that it is the percent reduction in viscosity provided by the use of the excipients described herein that is important. Regardless of the method used to measure viscosity, the percent reduction in viscosity of the excipient formulation versus the control formulation will be approximately the same at a given shear rate.

[0166] An amount or concentration of excipient effective to "reduce viscosity" means that the viscosity of the formulation in its final form for administration (in the case of a solution, or upon reconstitution with the intended amount of diluent in the case of a powder) is at least 5% less than the viscosity of other known viscosity-reducing agents, such as water, buffers, salts, etc., and control formulations, such as those exemplified herein. Control formulations that do not contain excipients can also be used, even if they are not implementable as therapeutic formulations, for example, due to hypotonicity.

[0167] Similarly, a "low viscosity" formulation is one that exhibits reduced viscosity compared to a control formulation.

[0168] A "pharmaceutical formulation" or "pharmaceutical composition" is a sterile composition of a pharmaceutically active drug, such as a biologically active protein, suitable for parenteral administration (including, but not limited to, intravenous, intramuscular, subcutaneous, aerosol, pulmonary, intranasal, or intrathecal) to a patient in need thereof, containing only pharmaceutically acceptable excipients, diluents, and other additives deemed safe by the Federal Drug Administration or other national regulatory agency. Pharmaceutical formulations include directly administrable liquid (e.g., aqueous) solutions and lyophilized powders that can be reconstituted into a solution by adding a diluent prior to administration. Specifically excluded from the scope of the term "pharmaceutical formulation" are compositions for topical administration to a patient, compositions for oral ingestion, and compositions for parenteral nutrition.

[0169] "Shelf life" refers to the storage period during which an active ingredient, such as a therapeutic protein, in a pharmaceutical formulation exhibits minimal degradation (e.g., about 5% to 10% degradation or less) when the pharmaceutical formulation is stored under specific storage conditions, e.g., at 2 to 8°C. Techniques for assessing degradation vary depending on the characteristics of the protein in the pharmaceutical formulation. Exemplary techniques include size exclusion chromatography (SEC)-HPLC, e.g., to detect aggregation; reversed-phase (RP)-HPLC, e.g., to detect protein fragmentation; ion exchange-HPLC, e.g., to detect changes in protein charge; mass spectrometry; fluorescence spectroscopy; circular dichroism (CD) spectroscopy; Fourier transform infrared spectroscopy (FT-IR); and Raman spectroscopy, which detects protein conformational changes. All of these techniques can be used alone or in combination to assess protein degradation in a pharmaceutical formulation and determine the shelf life of the formulation. Preferably, a pharmaceutical formulation exhibits no more than about 5% to about 10% increase in degradation (e.g., fragmentation, aggregation, or unfolding) over a two-year period when stored at 2 to 8°C.

[0170] "High molecular weight species" or "HMW species," in the context of a pharmaceutical formulation containing a therapeutic polypeptide, are therapeutic proteins that are larger than the original therapeutic polypeptide as determined by art-accepted assays. HMW species include oligomers of therapeutic polypeptides and aggregates of therapeutic polypeptides.

[0171] "Low molecular weight species" or "LMW species," in the context of a pharmaceutical formulation containing a therapeutic polypeptide, are polypeptides that are smaller than the original therapeutic polypeptide as determined by art-accepted assays. LMW species include fragments of the therapeutic polypeptide.

[0172] A "stable pharmaceutical formulation," "stable formulation," or "pharmaceutical formulation is stable" refers to a pharmaceutical formulation that exhibits limited increase in aggregation and / or a 5% or less decrease in biological activity when stored at about -30°C (or lower) to about 5°C to about 40°C for at least one month, or two months, or three months, or six months, or one year, or two years, or five years, or more, when compared to a control formulation sample. Formulation stability can be determined by one of skill in the art using any number of standard assays, including size exclusion HPLC (SEC-HPLC), cation exchange HPLC (CEX-HPLC), subvisible detection by light obscuration technology ("HIAC"), and / or visual inspection. Typically, higher storage temperatures result in shorter shelf-life formulations.

[0173] Techniques for assessing degradation vary depending on the identity of the protein in the pharmaceutical formulation. Exemplary techniques include size-exclusion chromatography (SEC)-HPLC, e.g., to detect aggregation; reversed-phase (RP)-HPLC, e.g., to detect protein fragmentation; ion-exchange-HPLC, e.g., to detect changes in protein charge; mass spectrometry; fluorescence spectroscopy; circular dichroism (CD) spectroscopy; Fourier transform infrared spectroscopy (FT-IR); and Raman spectroscopy, which detects protein conformational changes. All of these techniques can be used alone or in combination to assess protein degradation in pharmaceutical formulations and determine the shelf life of the formulations. The pharmaceutical formulations disclosed herein typically exhibit no more than about 2% to about 5% increase in degradation (e.g., fragmentation, aggregation, or unfolding) over a two-year period when stored at 2-8°C.

[0174] "Lyophilization," "lyophilized," and "freeze-drying" are processes in which the material to be dried is first frozen, and then the ice or freezing solvent is removed by sublimation under vacuum. To enhance the stability of the lyophilized product after storage, excipients can be included in the formulation before lyophilization.

[0175] "Diluent" refers to a substance that aids in the formulation and / or administration of and / or absorption of an active agent by a patient and that can be included in the compositions disclosed herein without causing significant adverse effects to the patient. An example of a diluent is water, which is preferably sterile and purified.

[0176] "Arginine salt" means a salt of arginine. Examples include arginine monohydrochloride (Arg-HCl), arginine acetate (Arg acetate), and arginine glutamate (Arg glutamate).

[0177] "N-acetylarginine" (NAR) refers to a molecule of formula 1. [ka]

[0178] "Polypeptide," also known as "protein," is used interchangeably. Exemplary polypeptides include antibodies, peptibodies, immunoglobulin-like proteins, non-antibody proteins, and non-immunoglobulin-like proteins. Analogs of native proteins, including polypeptides with altered glycosylation and unglycosylated (non-glycosylated) polypeptides, are contemplated for inclusion in the formulations of the present invention. "Protein analog" refers to an amino acid sequence that has insertions, deletions, or substitutions compared to the parent sequence while still substantially retaining the biological activity of the parent sequence as determined by biological assays known to those skilled in the art. Derivatives of native or analog polypeptides have been chemically modified, for example, to attach water-soluble polymers (e.g., PEGylated), radionuclides, or other diagnostic, targeting, or therapeutic moieties.

[0179] A "therapeutic protein" is a protein (or "therapeutic polypeptide", the terms are used interchangeably) that has at least one therapeutic (beneficial) effect in a patient.

[0180] Therapeutic proteins include antibodies and related molecules. An "antibody" or "immunoglobulin" refers to a tetrameric glycoprotein consisting of two heavy chains and two light chains, each containing a variable domain (V) and a constant domain (C). "Heavy chain" and "light chain" refer to substantially full-length canonical immunoglobulin light and heavy chains; the variable domains of the heavy and light chains (VL and VC) constitute the V region of the antibody and contribute to antigen binding and specificity. "Antibody" includes monoclonal, polyclonal, chimeric, human, and humanized antibodies. Light chains can be classified as kappa (κ) and lambda (λ) light chains. Heavy chains are typically classified as mu (μ), delta (δ), gamma (γ), alpha (α), or epsilon (ε), and antibody isotypes are defined as IgM, IgD, IgG, IgA, and IgE, respectively. IgG has several subclasses, including IgG1, IgG2, IgG3, and IgG4. IgM has subclasses, including IgM1 and IgM2. IgA is similarly further divided into subclasses, including IgA1 and IgA2. Within full-length light and heavy chains, the variable and constant regions are typically joined by a "J" region of about 12 or more amino acids, with heavy chains also including a "D" region of about 10 additional amino acids. The variable regions of each light / heavy chain pair typically form the antigen-binding site. A "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts.

[0181] "Antibody variants" include antibody fragments and antibody-like proteins with alterations in the structure of a canonical tetrameric antibody. Exemplary antibody variants include V regions with alterations in the constant region, or alternatively, V regions added to the constant region, optionally in a non-canonical manner. Examples include multispecific antibodies (e.g., bispecific antibodies, trispecific antibodies), antibody fragments capable of binding to antigen (e.g., Fab', F'(ab)2, Fv, single-chain antibodies, diabodies, diabodies), biparatopic peptides and recombinant peptides containing the above, so long as they exhibit the desired biological activity.

[0182] Multispecific antibodies target two or more antigens or epitopes. For example, "bispecific," "dual-specific," or "bifunctional" antibodies are hybrid antibodies with two different antigen-binding sites. Bispecific antibodies can be generated by various methods, including hybridoma fusion or Fab' fragment linkage (Kostelny et al. 1992, Songsivilai & Lachmann 1990, Wu & Demarest 2018). The two binding sites of a bispecific antibody each bind to a different epitope. Similarly, a trispecific antibody has three binding sites and binds to three epitopes. Several methods for generating trispecific antibodies are known and are under development (Wu & Demarest 2018, Wu et al. 2018). DART (dual affinity retargeting molecule) is also an example of a multispecific antibody.

[0183] BiTE® molecules: In some instances, the therapeutic protein is a bispecific T cell engager (BiTE) molecule. A BiTE molecule is a bispecific antibody construct or bispecific fusion protein containing two antibody binding domains (or targeting regions) linked to each other. One arm of the molecule is engineered to bind to a protein found on the surface of cytotoxic T cells, while the other arm is designed to bind to a specific protein found primarily on tumor cells. When both targets are engaged, the BiTE molecule forms a bridge between the cytotoxic T cells and the tumor cells, allowing the T cells to recognize and combat the tumor cells by injecting toxic molecules. For example, the tumor-binding arm of the molecule can be altered to create different BiTE antibody constructs that target different types of cancer. The term "binding domain" in reference to a BiTE molecule refers to the domain that (specifically) binds to / interacts with / recognizes a predetermined target epitope or predetermined target site on a target molecule (antigen). The structure and function of the first binding domain (tumor cell antigen recognition), and preferably also the structure and / or function of the second binding domain (cytotoxic T cell antigen), are based on the structure and / or function of an antibody (e.g., a full-length immunoglobulin molecule or an entire immunoglobulin molecule). For example, a BiTE molecule comprises a first binding domain characterized by the presence of three light chain CDRs (i.e., CDR1, CDR2, and CDR3 of the VL region) and three heavy chain CDRs (i.e., CDR1, CDR2, and CDR3 of the VH region). The second binding domain preferably also comprises the minimal structural requirements of an antibody that enable target binding. More preferably, the second binding domain comprises at least three light chain CDRs (i.e., CDR1, CDR2, and CDR3 of the VL region) and / or three heavy chain CDRs (i.e., CDR1, CDR2, and CDR3 of the VH region). It is envisaged that the first and / or second binding domains are generated or obtained by phage display or library screening methods, rather than by grafting onto a scaffold of CDR sequences from an existing (monoclonal) antibody. A binding domain may typically comprise an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH), but need not comprise both.An Fd fragment, for example, has two VH regions and often retains some of the antigen-binding function of the intact antigen-binding domain. Examples of (modified) antigen-binding antibody fragments include: (1) a Fab fragment, a monovalent fragment having the VL, VH, CL, and CH1 domains; (2) a F(ab')2 fragment, a bivalent fragment having two Fab fragments linked by a disulfide bridge at the hinge region; (3) a Fd fragment having two VH and CH1 domains; (4) an Fv fragment having the VL and VH domains of a single antibody arm; (5) a dAb fragment having a VH domain (Ward et al. 1989); (6) isolated complementarity-determining regions (CDRs); and (7) single-chain Fvs (scFvs), the latter being preferred (e.g., from an scFv library).

[0184] Antibody fragments include, for example, Fab, Fab', F(ab'), Fv, domain antibodies (dAbs), complementarity-determining region (CDR) fragments, CDR-grafted antibodies, single-chain antibodies (scFv), maxibodies (scFv-Fc), single-chain antibody fragments, chimeric antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies; chelating recombinant antibodies, tribodies or bibodies, intrabodies, nanobodies, small modular immunopharmaceuticals (SMIPs), antigen-binding domain immunoglobulin fusion proteins, single-domain antibodies (including camelized antibodies), VHH-containing antibodies, or variants or derivatives thereof, as well as polypeptides comprising at least a portion of an immunoglobulin sufficient to confer specific antigen binding on the polypeptide, e.g., the antigen-binding portion of an antibody comprising one, two, three, four, five, or six CDR sequences, so long as the antibody retains the desired binding activity.

[0185] References _______2014. Kollidon(R) - The Original. In BASF, ed. BASF:BASF Ausubel FM. 1987. Current protocols in molecular biology. Brooklyn, N.Y. Media, Pa.:Greene Pub. Associates; J. Wiley, order fulfillment. 2 volumes (loose-leaf) pp. Kamerzell TJ, Esfandiary R, Joshi SB, Middaugh CR, Volkin DB. 2011. Protein-excipient interactions:mechanisms and biophysical characterization applied to protein formulation development. Adv Drug Deliv Rev 63:1118-59 Kostelny SA, Cole MS, Tso JY. 1992. Formation of a bispecific antibody by the use of leucine zippers. J Immunol 148:1547-53 Powell MF, Nguyen T, Baloian L. 1998. Compendium of excipients for parenteral formulations. PDA J Pharm Sci Technol 52:238-311 Sambrook J, Russell DW. 2001. Molecular cloning :a laboratory manual. Cold Spring Harbor, N.Y.:Cold Spring Harbor Laboratory Press. Songsivilai S, Lachmann PJ. 1990. Bispecific antibody:a tool for diagnosis and treatment of disease. Clin Exp Immunol 79:315-21 Ward ES, Gussow D, Griffiths AD, Jones PT, Winter G. 1989. Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341:544-6 Wu X, Demarest SJ. 2018. Building blocks for bispecific and trispecific antibodies. Methods Wu X, Yuan R, Bacica M, Demarest SJ. 2018. Generation of orthogonal Fab-based trispecific antibody formats. Protein Eng Des Sel 31:249-56

[0186] Unless otherwise required by context, singular terms include pluralities and plural terms include the singular. In general, the nomenclature and techniques used in connection with cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are well known and commonly used in the art. Unless otherwise specified, the methods and techniques of the present invention are generally performed according to conventional methods well known in the art and as described in the general and more specific references cited and discussed throughout the specification. See, e.g., Ausubel et al. (1987 et seq.) and Sambrook et al. (2001) (Ausubel 1987, Sambrook & Russell 2001). Enzymatic reactions and purification procedures are performed according to manufacturer's instructions, as commonly accomplished in the art, or as described herein. The nomenclature used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques can be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

[0187] All identified patents and other publications are expressly incorporated herein by reference in their entirety for the purpose of describing and disclosing, for example, the methodology described in such publications that may be used in connection with the publications described.

Claims

1. 1. A composition comprising a concentration of an antibody and polyvinylpyrrolidone (PVP), wherein the PVP has a K value of 12 to 17, wherein the viscosity of the composition containing the PVP is lower than a composition comprising the same concentration of the antibody but without the PVP, and wherein the concentration of the antibody is greater than 70 mg / mL.

2. 10. The composition of claim 1, wherein the viscosity of the composition is 80 cP or less when measured at 25°C.

3. 3. The composition of claim 1 or 2, wherein the viscosity of the composition is read at 25°C and reported at a shear rate of 1,000 / s.

4. 2. The composition of claim 1, wherein the concentration of the antibody is greater than or equal to 140 mg / mL to 250 mg / mL.

5. 3. The composition of claim 1, wherein the PVP is present in a concentration of from 0.3% to 10%.

6. The composition of claim 1 , wherein the antibody is a monoclonal antibody (mAb).

7. The composition of claim 6 , wherein the monoclonal antibody (mAb) is an IgG monoclonal antibody.

8. 8. The composition of any one of claims 1 to 7, wherein the antibody is selected from the group consisting of a humanized antibody, a chimeric antibody, a multispecific antibody, a maxibody, a BiTE® molecule, a single chain antibody, a diabody, and a peptibody.

9. The composition of any one of claims 1 to 8, wherein the PVP has a K value of 12.

10. The composition of any one of claims 1 to 8, wherein the PVP has a K value of 17.

11. The composition of claim 1 or 2, further comprising arginine.

12. The composition of claim 11, wherein the arginine is N-acetylarginine.

13. 13. The composition of claim 12, wherein the N-acetylarginine is present at 10 mM.

14. 13. The composition of claim 11 or 12, wherein the arginine is a salt of arginine.

15. 15. The composition of claim 14, wherein the arginine is arginine monohydrochloride (Arg-HCl), the arginine monohydrochloride (Arg-HCl) is present at 67 mM, and the PVP is present at 1%.

16. 1. A method for reducing the viscosity of a pharmaceutical formulation comprising a therapeutic protein, the method comprising the step of combining the therapeutic protein with a viscosity-reducing concentration of PVP, wherein the PVP has a weight average molecular weight of 2,000 to 25,000 daltons, and the therapeutic protein has a concentration in the therapeutic formulation of at least 70 mg / mL.

17. 17. The method of claim 16, wherein the PVP has a K value of 12 to 17.

18. 17. The method of claim 16, wherein the PVP has a K value of 12.

19. 17. The method of claim 16, wherein the PVP has a K value of 17.

20. The method of any one of claims 16 to 19, wherein the therapeutic protein is a monoclonal antibody.

21. The method of any one of claims 16 to 19, wherein the viscosity of the composition is 80 cP or less.

22. 1. A lyophilized powder comprising a therapeutic protein and PVP, wherein the PVP has a K value of 12 to 17, the PVP being present at a weight:weight concentration effective to reduce viscosity after reconstitution with a diluent, and the PVP being present at a concentration of 100 μg / mg therapeutic protein to 1 mg / mg therapeutic protein.

23. 23. The lyophilized powder of claim 22, wherein the PVP has a K value of 12.

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