Viscosity reducing excipient compositions and low viscosity highly concentrated protein formulations comprising same

Abstract

Disclosed are an excipient composition for reducing the viscosity of a protein drug and a low-viscosity high-concentration protein composition preparation comprising the same, which have very low viscosity at a high protein concentration of about 20% or more, thereby being easily prepared, stored and used, and having a low protein drug viscosity and a low-viscosity high-concentration protein composition preparation comprising the same. And the method is particularly suitable for preparing injections for antibody therapy for subcutaneous and intramuscular administration. In particular, the high-concentration protein preparation can maintain a low viscosity even when stored at a low temperature, and thus can be used immediately after storage without separate preheating.

Description

Technical Field

[0001] The invention relates to an excipient composition for reducing the viscosity of a protein drug and a low-viscosity, high-concentration protein composition preparation containing the excipient composition. Background Art

[0002] Monoclonal antibodies (mAbs) have attracted much attention as therapeutic agents for various types of diseases, and therapeutic antibodies for a variety of diseases including cancer, neurodegenerative diseases, and immune diseases have been clinically approved and used, and are being continuously developed (Nature Reviews Immunology 6(5):343-357).

[0003] Conventional antibody drugs are generally administered by intravenous injection and are formulated into low-concentration IV dosage forms less than about 10 mg / mL for administration with an effective dose of generally about 100 mg to about 250 mg. However, the disadvantage of low-concentration IV dosage forms is that it takes a lot of time to administer to patients and requires frequent visits to the hospital (Canadian Oncology Nursing Journal 25(3):341-346.).

[0004] Recently, in order to overcome this shortcoming of low-concentration IV antibody preparations, people have actively worked on developing subcutaneous injection (SC) preparations of antibody drugs. In general, the advantages of subcutaneous preparations are small volume (1 to 1.5 mL), rapid administration, and especially self-injection (J Pharm Sci 93 (6): 1390-1402; Pharm Res. 2013, 30, 7). The effective dose of the antibody as an active ingredient is about 100 to about 250 mg, and the subcutaneous administration preparation of 1 to 1.5 mg / ml has a high concentration of about 100 mg / ml to about 300 mg / ml.

[0005] However, these high-concentration antibody preparations can induce protein-protein interactions between antibodies, form reversible or irreversible aggregates, induce immunogenicity, and exhibit high viscosity exceeding the pharmaceutically acceptable level (about 20 cP) (Industrial & Engineering Chemistry Research, 55 (43), 11225-11234). In addition, high-viscosity preparations may cause difficulties in protein preparation, especially in filtration.

[0006] It is reported that the high viscosity of high-concentration antibody preparations is due to various protein-protein interactions caused by the close distance between antibodies (Mol Pharmaceutics 12(1):127-139.), and a method for reducing the viscosity of antibody preparations by manipulating the amino acid sequence that triggers the interaction between antibodies has been reported (J. Pharm. Sci. 2013, 102, 8). However, this method has the limitations of being specific to a single antibody, taking a long time to develop, and not being generally applicable because the common problem of viscosity reduction exists in various protein drugs other than antibodies.

[0007] Therefore, studies have been conducted to reduce the viscosity of protein drugs using various excipients. A method of inhibiting the interaction between proteins by adding salts such as representative viscosity-reducing excipients, NaCl (Mol Pharmaceutics 12(1):127-139), calcium chloride or magnesium chloride (US 7,758,860), arginine hydrochloride, sodium thiocyanate, ammonium thiocyanate, ammonium sulfate, ammonium chloride, calcium chloride, zinc chloride or sodium acetate (US 7,666,413) has been reported. However, recent reports indicate that the addition of salts may not have an effect on the viscosity depending on the protein (Biophys J 106(8):1763-1770; Pharm Res 29(11):3102-3109), or may cause an increase in viscosity (Soft Matter 10(6):894-902).

[0008] It has also been reported that charged amino acids can be used as viscosity-reducing excipients in protein formulations. Specifically, arginine has been reported to reduce protein aggregation and viscosity (Biochemistry 44(12): 4919-4925). In addition, it has been reported that amino acids such as histidine and lysine can reduce the viscosity of protein formulations (Pharmaceutical Research, vol. 29, no. 11, June 13, 2012, p. 3182-3189).

[0009] Benzyl benzoate (Miller et al, Langmuir 26: 1067-1074, 2010), benzyl phenylacetate, ethanol, methyl ethyl ketone (Srinivasan et al, Pharm. Res. 30: 1749-1757) and the like have been reported as viscosity-reducing excipients using organic compounds. However, none of them has a sufficient viscosity-reducing effect at a level suitable for administration such as subcutaneous injection (about 20 cP or less).

[0010] FDA-approved antibody drugs for administration such as subcutaneous injection include Cuvitru, Hizentra, and Xembify, which contain 20% IgG antibodies. Cuvitru and Xembify use glycine (Cuvitru (250mM) and Xembify (160-260mM)) as a stabilizer, while Hizentra uses proline (250mM) as a stabilizer. These drugs need to be refrigerated, but they still show high viscosity after refrigeration, which is inconvenient to use.

[0011] In this context, the inventors have made strenuous efforts to develop a viscosity-reducing preparation for high-concentration protein drugs. Therefore, the inventors found that adding benzenesulfonic acid and camphorsulfonic acid to a conventional protein therapeutic composition containing proline or glycine greatly reduced the viscosity, and completed the present invention on this basis. Summary of the invention

[0012] An object of the present invention is to provide a viscosity-reducing excipient composition and a low-viscosity, high-concentration protein preparation comprising the composition.

[0013] Another object of the present invention is to provide a method for reducing the viscosity of a liquid protein-containing composition.

[0014] According to one aspect of the present invention, the above and other objects can be achieved by providing a pharmaceutical composition comprising at least one protein; glycine or proline; benzenesulfonic acid; and camphorsulfonic acid.

[0015] According to another aspect of the present invention, there is provided a viscosity reducing excipient composition comprising benzenesulfonic acid and camphorsulfonic acid, or pharmaceutically acceptable salts thereof.

[0016] According to another aspect of the present invention, there is provided a method for reducing the viscosity of a composition comprising a protein, comprising adding benzenesulfonic acid and camphorsulfonic acid as viscosity reducing excipients to a composition comprising at least one protein and glycine or proline. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The above and other objects, features and other advantages of the present invention will become more apparent through the following detailed description in conjunction with the accompanying drawings, in which:

[0018] Figure 1 is a graph showing the change in viscosity as a function of concentration between a 20% IgG base formulation containing proline and a formulation prepared by adding benzenesulfonic acid and / or camphorsulfonic acid to the 20% IgG base formulation containing proline.

[0019] Figure 2is a graph showing the change in viscosity as a function of concentration between a 20% IgG base formulation containing glycine and a formulation prepared by adding benzenesulfonic acid and / or camphorsulfonic acid to the 20% IgG base formulation containing glycine. DETAILED DESCRIPTION

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as understood by one of ordinary skill in the art to which the invention belongs. Generally speaking, the nomenclature used herein is well known and commonly used in the art.

[0021] Protein therapy is a targeted therapy for specific diseases, and therapeutic proteins for various diseases such as cancer and immune-related diseases are developed and used. At present, protein therapeutics are mostly administered by intravenous injection in low-concentration preparations, but high-concentration preparations for effective subcutaneous or intramuscular administration are being actively developed recently. However, high-concentration protein compositions have high viscosity due to protein interactions, which brings difficulties to manufacturing, management and administration, and causes discomfort such as pain to patients.

[0022] To date, various excipients and combinations have been reported for various low-viscosity, high-concentration protein formulations, and all have shown slight reductions in viscosity. Still, only a few excipient combinations have shown sufficient viscosity reduction effects for clinical use. In particular, previously reported viscosity-reducing excipients have shown an increase in formulation viscosity when stored at low temperatures, which is essential for the preservation of protein drugs. Therefore, preheating before use is essential. In the process, the viscosity-reducing excipients may, disadvantageously, become deformed or contaminated.

[0023] In one embodiment of the present invention, it was found that when a combination of benzenesulfonic acid and camphorsulfonic acid was added to a conventional protein therapeutic composition containing proline or glycine, the viscosity of the protein formulation was significantly reduced, and in particular, the viscosity was low enough for immediate administration even under low temperature refrigeration conditions.

[0024] In one aspect, the present invention provides a pharmaceutical composition comprising at least one protein; glycine or proline; benzenesulfonic acid; and camphorsulfonic acid.

[0025] As used herein, the term "protein" refers to an amino acid polymer that forms a polypeptide having a sufficient length to produce a tertiary structure connected by peptide bonds. In general, proteins are divided into high molecular weight proteins of 100 kDa or higher and low molecular weight proteins of less than 100 kDa. In some embodiments of the present invention, the protein may have at least one biological effect, preferably, the protein may be a "therapeutic protein" that has a preventive, ameliorative or therapeutic effect on at least one disease.

[0026] Preferably, the protein may be an antigen binding protein capable of specifically binding to a target substance or antigen.

[0027] As used herein, the term "antigen binding protein" refers to a protein that can recognize an antigen and specifically bind to it. In a broad sense, non-limiting examples of antigen binding proteins include antibodies or antigen-binding fragments thereof, antibody-like proteins (ALPs) such as diabodies, pentabodies, repebodies or affimers, and antigen-binding peptides such as C7 peptides.

[0028] In some embodiments of the present invention, the protein is preferably an antibody or a binding fragment thereof.

[0029] The term "antibody" refers to a protein that is capable of neutralizing, blocking, inhibiting, eliminating, reducing or interfering with a biological activity.

[0030] The antibodies of the present invention include, but are not limited to, monoclonal antibodies, multispecific antibodies, human antibodies, humanized antibodies, chimeric antibodies, single-chain FV (scFV), single-chain antibodies, Fab fragments, F(ab') fragments, disulfide bond Fv (sdFV), anti-idiotypic (anti-Id) antibodies, epitope-binding fragments of such antibodies, etc.

[0031] The term "monoclonal antibody" refers to an identical antibody obtained from a population of substantially homogeneous antibodies, ie, each antibody comprising the population is exclusive of any trace naturally occurring mutations that it may contain. Monoclonal antibodies are highly specific and are therefore produced against a single antigenic site.

[0032] The term "epitope" refers to a protein determinant to which an antibody can specifically bind. An epitope is usually composed of a group of chemically active surface molecules, such as amino acids or sugar side chains, and generally has not only specific three-dimensional structural characteristics, but also specific charge characteristics. The difference between a three-dimensional epitope and a non-three-dimensional epitope is that the bonds of the former will break in the presence of denaturing solvents, while the bonds of the latter will not break.

[0033] A whole antibody has a structure of two full-length light chains and two full-length heavy chains, and each light chain is bound to a heavy chain by a disulfide bond. Whole antibodies include IgA, IgD, IgE, IgM and IgG, of which IgG can include IgG1, IgG2, IgG3 and IgG4 subtypes.

[0034] Heavy chain constant regions include gamma (γ), mu (μ), alpha (α), delta (δ), and epsilon (ε), which are further divided into gamma 1 (γ1), gamma 2 (γ2), gamma 3 (γ3), gamma 4 (γ4), alpha1 (α1), and alpha 2 (α2). Light chain constant regions include kappa (κ) and lambda (λ).

[0035] The term "antigen-binding fragment of an antibody" or "antibody fragment" refers to a fragment having an antigen-binding function, including Fab, F(ab'), F(ab')2, Fv, etc. Among antibody fragments, Fab refers to a structure including the variable regions of each of the heavy chain and the light chain, the constant region of the light chain, and the first constant region (CH1) of the heavy chain, each having one antigen-binding site. Fab' differs from Fab in that it also includes a hinge region containing at least one cysteine ​​residue at the C-terminus of the heavy chain CH1 domain. F(ab')2 is formed by disulfide bonds between the cysteine ​​residues in the hinge region of Fab'. Fv is the smallest antibody fragment having only the heavy chain variable region and the light chain variable region. A two-chain Fv is a fragment in which the heavy chain variable region and the light chain variable region are linked by non-covalent bonds, and a single-chain Fv (scFv) is a fragment in which the heavy chain variable region and the light chain variable region are usually linked by covalent bonds via a peptide linker therebetween, or are directly linked at the C-terminus to form a dimer-like structure (such as a two-chain Fv). Such antibody fragments can be obtained using proteases (eg, Fab can be obtained by restricting the cleavage of an intact antibody with papain, and F(ab')2 fragments can be obtained by restricting the cleavage of an intact antibody with pepsin), and can be produced using genetic recombination techniques.

[0036] As used herein, the term "Fv fragment" is an antibody fragment that contains the recognition and binding sites of an intact antibody. Such regions include a dimer consisting of one heavy chain variable domain and one light chain variable domain, the two domains being essentially tightly covalently linked together, such as by scFv.

[0037] As used herein, the term "Fab" fragment comprises the variable domain and constant domain of the light chain and the variable domain and first constant domain (CH1) of the heavy chain. F(ab')2 antibody fragments typically comprise a pair of Fab fragments covalently linked near their carboxyl termini by hinge cysteines therebetween.

[0038] As used herein, "single-chain Fv" or "scFv" antibody fragments include the VH and VL domains of an antibody, wherein these domains are included in a single polypeptide chain. The Fv polypeptide may also include a polypeptide linker between the VH and VL domains to allow the scFv to form a target structure for antigen binding.

[0039] For example, in some embodiments of the invention, the antibody is in the form of an Fv (e.g., scFv) or a complete antibody. In addition, the heavy chain constant region can be selected from gamma (γ), mu (μ), alpha (α), delta (δ), and epsilon (ε) isotypes. For example, the constant region can be γ1 (IgG1), γ3 (IgG3), or γ4 (IgG4). The light chain constant region can be κ or λ.

[0040] As used herein, the term "heavy chain" encompasses full-length heavy chains, including variable domains (VH), which include amino acid sequences with variable region sequences sufficient to confer antigen specificity and three constant domains (CH1, CH2, and CH3), and fragments thereof. As used herein, the term "light chain" encompasses full-length light chains, including variable domains (VL), which include amino acid sequences with variable region sequences sufficient to confer specificity to antigens, and constant domains (CL), and fragments thereof.

[0041] "Chimeric" antibodies (immunoglobulins) in which a portion of the heavy and / or light chain is identical or homologous to the corresponding sequence of an antibody from a particular species or belonging to a particular antibody class or subclass, while the remaining chain or chains are identical or homologous to the corresponding sequence of an antibody from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies that exhibit the desired biological activity.

[0042] As used herein, the term "antibody variable domain" refers to the light chain region and heavy chain region of an antibody molecule comprising the amino acid sequence of the complementary determining region (CDR; i.e., CDR1, CDR2, and CDR3) and the framework region (FR). VH refers to the variable domain of the heavy chain. VL refers to the variable domain of the light chain.

[0043] The term "complementarity determining region" (CDR, i.e., CDR1, CDR2 and CDR3) refers to the amino acid residues in the variable domain of an antibody that are essential for antigen binding. Each variable domain usually has three CDR regions, referred to as CDR1, CDR2 and CDR3.

[0044] Non-human (e.g., murine) antibodies are chimeric antibodies containing minimal sequence from non-human immunoglobulins. Humanized antibodies are human immunoglobulins (recipient antibodies) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or non-human primate having the desired specificity, affinity, and capacity.

[0045] In some embodiments of the present invention, the protein may be an antibody, preferably a monoclonal antibody, and more preferably an IgG antibody. In some embodiments of the present invention, the antibody may be various therapeutic antibodies that are commercially available or under development in the art.

[0046] In some embodiments of the invention, the protein can be a therapeutic protein other than an antibody. For example, the protein can be an enzyme, a fusion protein, a PEGylated protein, a vaccine, a biologically active protein, or a protein mixture.

[0047] As used herein, the term "enzyme" refers to a protein or a functional fragment thereof that catalyzes the biochemical conversion of a molecule into a desired product.

[0048] As used herein, the term "fusion protein" refers to a protein produced by two different genes encoding two independent proteins. Fusion proteins are usually produced by recombinant DNA techniques known to those skilled in the art. Two proteins (or protein fragments) exhibit properties derived from the two parent proteins to which they are covalently fused.

[0049] In some embodiments of the present invention, the protein may be a conjugate with another compound. For example, the conjugate may be an antibody-drug conjugate, but is not limited thereto.

[0050] In some embodiments of the present invention, proteins may be included as "active ingredients" having medical and / or pharmacological activities.

[0051] In some embodiments of the present invention, the amount of the protein included may be a pharmaceutically active amount. For example, a pharmaceutically active single dose of an antibody therapeutic may be about 150 mg to about 250 mg, but is not limited thereto.

[0052] The pharmaceutical compositions of the present invention may contain high concentrations of protein.

[0053] In some embodiments of the invention, the concentration of the protein is about 1 mg / mL or more, about 10 mg / mL or more, or about 50 mg / mL or more, more preferably about 150 mg / mL or more, and more preferably about 200 mg / mL or more.

[0054] In some embodiments of the invention, the concentration of the protein is from about 10 mg / mL to about 5,000 mg / mL, preferably from about 50 mg / mL to about 2,500 mg / mL, more preferably from about 100 mg / mL to about 1,000 mg / mL, more preferably from about 100 mg / mL to about 500 mg / mL, and most preferably from about 150 mg / mL to about 500 mg / mL, but is not limited thereto.

[0055] In some embodiments of the present invention, the concentration of protein is about 1 w / v% to about 70 w / v%, more preferably about 5 w / v% to about 50 w / v%, more preferably about 7 w / v% to about 35 w / v%, more preferably about 10 w / v% to about 25 w / v%, but is not limited thereto.

[0056] In one embodiment of the present invention, the protein is included at a concentration of about 200 mg / mL (about 20% w / v), but is not limited thereto.

[0057] In some embodiments of the present invention, glycine or proline, benzenesulfonic acid and camphorsulfonic acid may be included as excipients.

[0058] As used herein, the term "excipient" refers to an additive other than an active ingredient that has one or more physical / chemical effects.

[0059] In some embodiments of the present invention, glycine or proline may be included as a stabilizer. As used herein, the term "stabilizer" refers to an additive used to impart physical stability or chemical stability to a pharmaceutical composition. For example, after storage at 4°C for 24 months, or under equivalent solution conditions at elevated temperatures, such as after storage at 40°C for 1 month, at least about 95% of the bioactive protein molecules may retain the biological activity of the formulation, but is not limited thereto.

[0060] In some embodiments of the present invention, glycine or proline, in addition to stabilizing protein activity, may also affect one or more physical and / or chemical properties of the composition, such as viscosity reduction and isotonicity.

[0061] In some embodiments of the present invention, the concentration of the glycine or proline included may be about 10 mM to about 1,000 mM, preferably about 50 mM to about 500 mM, more preferably about 100 mM to about 400 mM, more preferably about 150 mM to about 300 mM. In one embodiment of the present invention, a composition comprising 250 mM glycine or proline is prepared, but is not limited thereto.

[0062] In one embodiment of the present invention, it was found that when benzenesulfonic acid and camphorsulfonic acid were further applied, the viscosity of the composition was significantly reduced, and in particular, the composition had a low viscosity even under low temperature refrigeration conditions of 4°C.

[0063] As used herein, the term "benzenesulfonic acid" is an aromatic sulfonic acid having the structure shown in Formula I below.

[0064] [Formula I]

[0065]

[0066] Benzenesulfonic acid can be produced by various conventionally known methods, and is generally produced by sulfonating benzene using sulfuric acid (Otto Lindner, Lars Rodefeld (2005). "Benzenesulfonic Acids and Their Derivatives". Ullmann's Encyclopedia of Industrial Chemistry.), but is not limited thereto.

[0067] In some embodiments of the present invention, the benzenesulfonic acid included may be benzenesulfonate, which is a pharmaceutically acceptable salt thereof. The benzenesulfonic acid included may be a salt of an alkali metal or alkaline earth metal such as sodium, potassium, lithium, calcium or magnesium, but is not limited thereto.

[0068] In some embodiments of the present invention, the benzenesulfonic acid (BA) is included in a concentration of about 5 mg / mL to about 30 mg / mL, preferably about 10 mg / mL to about 27.5 mg / mL, more preferably about 10 mg / mL to about 25 mg / mL, and most preferably about 10 mg / mL to about 20 mg / mL.

[0069] As used herein, the term "camphorsulfonic acid (CA)" is an aromatic sulfonic acid represented by 1S-(+)-10-camphorsulfonic acid, having the structure shown in the following Formula II.

[0070] [Formula II]

[0071]

[0072] Camphorsulfonic acid can be prepared by various methods known in the art, and is generally prepared by sulfonation of a methyl group, preferably forming an olefin from the methyl group, followed by sulfonation of the olefin and rearrangement of the semi-pinacol (Bruckner, Reinhard (2002). Advanced organic chemistry: reaction mechanisms.), but is not limited thereto.

[0073] In some embodiments of the present invention, the camphorsulfonic acid may be a camphorsulfonate, which is a pharmaceutically acceptable salt thereof. The camphorsulfonic acid may be a salt of an alkali metal or alkaline earth metal such as sodium, potassium, lithium, calcium or magnesium, but is not limited thereto.

[0074] In some embodiments of the present invention, the camphorsulfonic acid (CA) is included in a concentration of about 5 mg / mL to about 30 mg / mL, preferably about 10 mg / mL to about 27.5 mg / mL, more preferably about 10 mg / mL to about 25 mg / mL, and most preferably about 10 mg / mL to about 20 mg / mL.

[0075] It is found in the examples of the present invention that the combination of benzenesulfonic acid and camphorsulfonic acid has a significant viscosity reducing effect compared with single benzenesulfonic acid or camphorsulfonic acid at the same total concentration.

[0076] In some embodiments of the present invention, the total concentration of benzenesulfonic acid and camphorsulfonic acid is about 10 mg / mL to about 60 mg / mL, preferably about 20 mg / mL to about 50 mg / mL, more preferably about 20 mg / mL to about 40 mg / mL, and most preferably about 30 mg / mL to 40 mg / mL.

[0077] In some embodiments of the present invention, the weight ratio of the benzenesulfonic acid to the camphorsulfonic acid may be 1:0.5 to 1:2.5, preferably 1:1 to 1:2.5, more preferably 1:2.0, 1:1 or 1:0.75.

[0078] In some embodiments of the present invention, the benzenesulfonic acid and the camphorsulfonic acid may be included as viscosity reducing excipients.

[0079] As used herein, the term "viscosity reducing excipient" or "viscosity reducing agent" refers to a compound that acts to reduce the viscosity of a solution containing the viscosity reducing agent compared to a solution that does not contain the viscosity reducing agent.

[0080] As used herein, the term "viscosity" refers to the resistance of a fluid (liquid or gas) to flow or to the movement of adjacent parts relative to each other. The inverse of viscosity can be expressed as flowability. Viscosity can be related to the concept of shear force, which can be understood as the effect of different layers of a fluid exerting shear forces on each other or on another surface when moving relative to each other. Viscosity can be expressed in Ns / m 2 It is also called Pascal*second (Pa*s). Viscosity can be "kinematic viscosity" or "absolute viscosity". "Viscosity" used in this article is generally understood to be "absolute viscosity".

[0081] Kinematic viscosity is a measure of the rate at which momentum is transferred through a fluid. Kinematic viscosity is measured in units of stokes (St). Kinematic viscosity can be measured by the resistance of a fluid to flow under the influence of gravity. When two fluids of equal volume and different viscosities are injected into the same capillary viscometer and allowed to flow under the influence of gravity, the fluid with the higher viscosity will take longer to flow through the capillary than the fluid with the lower viscosity. For example, if one fluid takes 200 seconds (s) to complete its flow and the other fluid takes 400 seconds (s) to complete its flow, the second fluid may be twice as viscous as the first fluid in terms of kinematic viscosity. The units of kinematic viscosity can be expressed as length 2 / hour. Kinematic viscosity is usually expressed in centistokes (cSt), and the SI unit of kinematic viscosity is cm 2 / s, which is equal to 1St.

[0082] As used herein, the term "absolute viscosity" is used interchangeably with "dynamic viscosity" or "simple viscosity" and refers to the product of kinematic viscosity and fluid density. Absolute viscosity is expressed in centipoise (cP). The SI unit of absolute viscosity is milliPascal-second (mPa-s), where 1 cP = 1 mPa*s.

[0083] In some embodiments of the present invention, viscosity can be measured by various methods, viscometers and rheometers known in the art. For example, a glass capillary viscometer, a Stormer viscometer, a vibration viscometer, etc. can be used to measure viscosity, but it is not limited thereto.

[0084] In some embodiments of the present invention, the pharmaceutical composition may have a low viscosity. In some embodiments of the present invention, the viscosity of the pharmaceutical composition may be 50 cP or less, preferably about 40 cP or less, about 35 cP or less, or about 30 cP or less, more preferably about 20 cP or less, or about 15 cP or less, but not limited thereto.

[0085] In some embodiments of the invention, the viscosity of the pharmaceutical composition is reduced by at least 10%, at least 20%, at least 30%, at least 40% or at least 50% compared to a formulation not comprising benzenesulfonic acid and camphorsulfonic acid as viscosity reducing excipients.

[0086] In some embodiments of the invention, the viscosity of the pharmaceutical composition can vary depending on temperature. In one embodiment of the invention, it was found that when benzenesulfonic acid and camphorsulfonic acid were included as viscosity-reducing excipients, a substantial reduction in viscosity was obtained at about 4°C, which is the storage temperature of a typical protein formulation.

[0087] In some embodiments of the present invention, the viscosity of the pharmaceutical composition at about 4°C can be 50 cP or less, preferably about 40 cP or less at about 4°C, about 35 cP or less at about 4°C, or about 30 cP or less at about 4°C, more preferably about 20 cP or less at about 4°C, or about 15 cP or less at about 4°C, but is not limited thereto.

[0088] In some embodiments of the invention, the viscosity of the pharmaceutical composition is 50 cP or less, preferably about 40 cP or less, about 35 cP or less, or about 30 cP or less, more preferably about 20 cP or less, or about 15 cP or less, even when stored at about 4° C. for at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, or at least 1 day.

[0089] The pharmaceutical composition of the present invention can be used to prepare high-concentration protein preparations.

[0090] The pharmaceutical composition of the present invention may contain, in addition to proline or glycine, benzenesulfonic acid and camphorsulfonic acid, suitable carriers, excipients and diluents commonly used in pharmaceutical compositions.

[0091] Examples of carriers, excipients or diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, etc. The formulation of the composition may be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants or surfactants.

[0092] In particular, pharmaceutical excipients for liquid protein formulations are well known to those skilled in the art. Non-limiting examples include: liquid solvents or cosolvents; sugars or sugar alcohols such as mannitol, trehalose, sucrose, sorbitol, fructose, maltose, lactose or dextran; surfactants such as 20, 60 or 80 (polysorbate 20, 60, or 80); buffers; preservatives such as benzalkonium chloride, benzethonium chloride, tertiary ammonium salts and chlorhexidine diacetate; carriers such as polyethylene glycol (PEG); antioxidants such as ascorbic acid, sodium bisulfite and methionine; chelating agents such as EDTA or citric acid; or biodegradable polymers such as water-soluble polyesters; cryoprotectants; lyoprotectants; bulking agents; and stabilizers. In addition, in Remington: "The Science and Practice of Pharmacy" 20 thOther pharmaceutically acceptable carriers, excipients, or stabilizers described in 2000 (Alfonso R Gennaro, Ed., Lippincott Williams & Wilkins (2000)) may be included in the protein formulations described herein as long as they do not adversely affect the desired properties of the formulation.

[0093] Pharmaceutical composition according to the present invention can utilize inactive organic or inorganic carrier in pharmacology to make suitable preparation.That is, when preparation is tablet, coated tablet, dragee or hard capsule, it can comprise lactose, sucrose, starch or its derivative, talcum, calcium carbonate, gelatin, stearic acid or their salt.In addition, when preparation is soft capsule, it may comprise vegetable oil, wax, fat or semisolid or liquid polyol.In addition, when preparation is solution or syrup form, it may comprise water, polyol, glycerine, vegetable oil etc.

[0094] The pharmaceutical composition according to the present invention may further contain, in addition to the carrier, a preservative, a stabilizer, a wetting agent, an emulsifier, a solubilizer, a flavoring agent, a colorant, an osmotic pressure regulator, an antioxidant, and the like.

[0095] The pharmaceutical composition according to the present invention can be administered in a pharmaceutically effective amount, and "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio, which is applicable to all medical treatments, and the effective dosage level can be determined based on various factors including the patient's disease type, the severity of the disease, the activity of the drug, the patient's sensitivity to the drug, the administration time, the administration route, the excretion rate, the treatment period, the drugs used at the same time, and other factors known in the pharmaceutical field. The pharmaceutical composition of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered in a single dose or multiple doses. Taking these factors into account, it is important to administer the minimum amount sufficient to achieve the maximum therapeutic effect without side effects, and the amount can be easily determined by those skilled in the art.

[0096] In some embodiments of the present invention, the pharmaceutical composition can be administered by a variety of routes. For example, the pharmaceutical composition can be administered to a patient subcutaneously, intramuscularly, intraperitoneally, intracerebrospinal fluid, intravenously, intraarticularly, intrasynovially, intrathecally, orally, or topically, or by inhalation, most preferably subcutaneously or intramuscularly, but not limited thereto. When the pharmaceutical composition of the present invention is administered subcutaneously, an injection device (e.g., INJECT-EASE TM and GENJECT TM ), injection pens (e.g., GENPEN TM ), needle-free devices (e.g., MEDIJECTOR TM and BIOJECTOR TM) or subcutaneous patch delivery system.

[0097] The pharmaceutical preparation according to the present invention can be provided for administration in the form of being stored in a glass ampoule or a plastic container, or in a ready-to-use form such as a prefilled syringe. Specifically, when the preparation is administered by subcutaneous injection or intramuscular injection, it can be subdivided into a volume of about 1 mL to 10 mL, but is not limited thereto, and if necessary, it can be administered after dilution with an appropriate solution such as physiological saline or glucose injection.

[0098] The pharmaceutical composition of the present invention exhibits high stability even at a low temperature of 4° C. and low viscosity that allows immediate administration, and thus can be administered immediately even after refrigeration.

[0099] After being refrigerated separately, the pharmaceutical composition of the present invention can be administered immediately without preheating.

[0100] The pharmaceutical composition of the present invention can be administered immediately without requiring pre-administration procedures such as preheating or dilution, thereby avoiding contamination or modification and / or inactivation of proteins (preferably as active ingredients) that may occur during pre-administration procedures.

[0101] In another aspect, the present invention relates to a viscosity reducing excipient comprising benzenesulfonic acid and camphorsulfonic acid.

[0102] In some embodiments of the present invention, the benzenesulfonic acid included may be benzenesulfonate, which is a pharmaceutically acceptable salt thereof. The benzenesulfonic acid included may be an alkali metal or alkaline earth metal such as sodium, potassium, lithium, calcium or magnesium, but is not limited thereto.

[0103] In some embodiments of the present invention, the benzenesulfonic acid (BA) is included in a concentration of about 5 mg / mL to about 30 mg / mL, preferably about 10 mg / mL to about 27.5 mg / mL, more preferably about 10 mg / mL to about 25 mg / mL, and most preferably about 10 mg / mL to about 20 mg / mL.

[0104] In some embodiments of the present invention, the camphorsulfonic acid may be a camphorsulfonate, which is a pharmaceutically acceptable salt thereof. The camphorsulfonic acid may be a salt of an alkali metal or alkaline earth metal such as sodium, potassium, lithium, calcium or magnesium, but is not limited thereto.

[0105] In some embodiments of the present invention, the camphorsulfonic acid (CA) is included in a concentration of about 5 mg / mL to about 30 mg / mL, preferably about 10 mg / mL to about 27.5 mg / mL, more preferably about 10 mg / mL to about 25 mg / mL, and most preferably about 10 mg / mL to about 20 mg / mL.

[0106] In some embodiments of the present invention, the total concentration of the benzenesulfonic acid and the camphorsulfonic acid is about 10 mg / mL to about 60 mg / mL, preferably about 20 mg / mL to about 50 mg / mL, more preferably about 20 mg / mL to about 60 mg / mL, about 40 mg / mL, and most preferably about 30 mg / mL to 40 mg / mL.

[0107] In some embodiments of the present invention, the weight ratio of the benzenesulfonic acid to the camphorsulfonic acid may be 1:0.5 to 1:2.5, preferably 1:1 to 1:2.5, more preferably 1:2.0, 1:1 or 1:0.75.

[0108] In some embodiments of the invention, viscosity reducing excipients may be used to reduce the viscosity of high concentration protein formulations.

[0109] In some embodiments of the present invention, the high concentration protein preparation may be a liquid preparation.

[0110] In some embodiments of the invention, the concentration of the protein in the high concentration protein preparation is about 1 mg / mL or higher, about 10 mg / mL or higher, or about 50 mg / mL or higher, more preferably about 150 mg / mL or higher, and more preferably about 200 mg / mL or higher.

[0111] In some embodiments of the invention, the concentration of the protein is about 10 mg / mL to about 5,000 mg / mL, preferably about 50 mg / mL to about 2,500 mg / mL, more preferably about 100 mg / mL to about 1,000 mg / mL, more preferably about 100 mg / mL to about 500 mg / mL, and most preferably about 150 mg / mL to about 500 mg / mL, but is not limited thereto.

[0112] In some embodiments of the present invention, the concentration of the protein is about 1 w / v% to about 70 w / v%, more preferably about 5 w / v% to about 50 w / v%, more preferably about 7 w / v% to about 35 w / v%, more preferably about 10 w / v% to about 25 w / v%, but is not limited thereto.

[0113] In one embodiment of the present invention, the concentration of the protein is about 200 mg / mL (about 20% w / v), but is not limited thereto.

[0114] In some embodiments of the present invention, the viscosity reducing excipient may further comprise proline or glycine.

[0115] In some embodiments of the present invention, the concentration of the glycine or proline included may be about 10 mM to about 1,000 mM, preferably about 50 mM to about 500 mM, more preferably about 100 mM to about 400 mM, more preferably about 150 mM to about 300 mM. In one embodiment of the present invention, a composition comprising 250 mM glycine or proline is prepared, but is not limited thereto.

[0116] In another aspect, the present invention relates to a method of reducing the viscosity of a composition comprising a protein, which comprises adding benzenesulfonic acid and camphorsulfonic acid as viscosity reducing excipients to a composition comprising at least one protein and glycine or proline.

[0117] [Example]

[0118] The present invention will be described in more detail below in conjunction with examples. However, it will be apparent to those skilled in the art that these examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention.

[0119] Example 1. Preparation of high-concentration protein preparations

[0120] To confirm the viscosity reducing effect of the combination of benzenesulfonic acid and camphorsulfonic acid in a high-concentration protein formulation, benzenesulfonic acid (BA) and / or camphorsulfonic acid (CA) were added as viscosity reducing excipients (Table 2) to a base formulation containing 20% ​​human IgG (human immunoglobulin G, green cross) and proline or glycine (Table 1) to prepare a liquid composition.

[0121] [Table 1]

[0122] Basic preparations

[0123] Element concentration protein 20% IgG (w / v) Glycine or Proline 250mM Polysorbate 80 0.02%(w / v)

[0124] [Table 2]

[0125] Viscosity reducing excipients

[0126]

[0127] Example 2. Confirmation of viscosity reduction effect of high concentration protein preparation under low temperature refrigeration conditions

[0128] Example 2-1: Viscosity measurement method

[0129] To measure the viscosity of the proline-based 20% IgG formulations, 500 μL of each reduced viscosity formulation sample shown in Table 2 was measured under refrigerated conditions at 4°C using the RheoSense mVROC according to the developer's protocol.

[0130] Example 2-2: Viscosity reduction effect of a proline-based 20% IgG formulation

[0131] The viscosity of the 20% IgG protein preparation containing proline after refrigeration at 4°C is shown in Table 3 below ( Figure 1 ).

[0132] [Table 3]

[0133] Viscosity of proline-based formulations (4°C)

[0134] Preparation (Proline) Viscosity(cP) Basic preparations 29.79 BA 10 (Proline) 29.548 CA 10 (Proline) 28.904 BA 20 (Proline) 28.007 CA 20 (proline) 30.722 BA 5+CA 5 (proline) 28.397 BA 10+CA 10 (proline) 23.332 BA 15+CA 15 (proline) 21.523 BA 10+CA 20 (proline) 14.834 BA 20+CA 20 (proline) 14.523 BA 25+CA 25 (proline) 21.748 BA 12.5+CA 27.5 (proline) 25.486 BA 30+CA 30 (proline) 29.745

[0135] As shown in Table 3, when benzenesulfonic acid or camphorsulfonic acid is added alone, no matter how its concentration is, there is no effect on viscosity. However, when benzenesulfonic acid and camphorsulfonic acid are applied in combination, viscosity decreases. Specifically, when the addition amount of benzenesulfonic acid and camphorsulfonic acid is 10mg / mL, 15mg / mL or 25mg / mL (BA 10+CA 10, or BA 25+CA 25), or when a combination of 12.5mg / mL of benzenesulfonic acid and 27.5mg / mL of camphorsulfonic acid is added (BA 12.5+CA 27.5), a viscosity reduction effect of about 20% to 30% can be obtained under low temperature refrigerated storage conditions, and when a combination of 10mg / mL of benzenesulfonic acid and 20mg / mL of camphorsulfonic acid is added (BA 10+CA20), or 20mg / mL of benzenesulfonic acid and camphorsulfonic acid (BA20+CA20), the viscosity decreases by more than 50%.

[0136] On the other hand, when the addition amount of benzenesulfonic acid and camphorsulfonic acid was 30 mg / mL or more respectively, the viscosity reducing effect of the basic preparation was not obtained, which means that the viscosity reducing effect of benzenesulfonic acid and camphorsulfonic acid does not simply increase with increasing concentration, but is caused by the specific concentration and concentration ratio defined in the present invention.

[0137] Example 2-3: Viscosity reduction effect of a glycine-based 20% IgG formulation

[0138] The viscosity of the 20% IgG protein preparation containing glycine after refrigeration at 4°C is shown in Table 4 below ( Figure 2 ).

[0139] [Table 4]

[0140] Viscosity of glycine-based formulations (4°C)

[0141] Preparation (Glycine) Viscosity(cP) Basic preparations 38.557 BA20+CA15(Glycine) 26.154

[0142] When glycine is included in the formulation, the combination of benzenesulfonic acid and camphorsulfonic acid can provide about 33% viscosity reduction compared to the formulation without glycine.

[0143] Industrial Applicability

[0144] The excipient composition for reducing the viscosity of a protein drug according to the present invention and the low-viscosity high-concentration protein preparation containing the excipient composition have very low viscosity even when the protein concentration is about 20% or higher, so that they can be easily manufactured, stored and used, and are particularly suitable for preparing antibody therapeutic injections for subcutaneous and intramuscular administration. In particular, the high-concentration protein preparation of the present invention can maintain low viscosity even when stored at low temperatures, so it can be used immediately after storage without separate preheating.

[0145] Although the specific configuration of the present invention has been described in detail, it will be appreciated by those skilled in the art that this description is only for the purpose of illustration to set forth the preferred embodiment and should not be interpreted as limiting the scope of the present invention. Therefore, the true scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition comprising: at least one protein; glycine or proline; Benzenesulfonic acid; and camphorsulfonic acid.

2. The pharmaceutical composition according to claim 1, wherein The protein is an antibody or an antigen-binding fragment thereof.

3. The pharmaceutical composition according to claim 1, wherein The protein is included in a concentration of 10% to 25% (w / v).

4. The pharmaceutical composition according to claim 1, wherein The glycine or protein is included at a concentration of 150 mM to 250 mM.

5. The pharmaceutical composition according to claim 1, wherein The benzenesulfonic acid and camphorsulfonic acid are each independently included in a concentration of 10 mg / mL to 27.5 mg / mL.

6. The pharmaceutical composition according to claim 1, wherein The total concentration of the benzenesulfonic acid and the camphorsulfonic acid is 20 mg / mL to 50 mg / mL.

7. The pharmaceutical composition according to claim 1, wherein The benzenesulfonic acid and the camphorsulfonic acid are included in a weight ratio of 1:0.5 to 1:2.

5.

8. The pharmaceutical composition according to claim 1, wherein The absolute viscosity of the pharmaceutical composition at 4° C. is less than 25 cP.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein The pharmaceutical composition is administered by subcutaneous injection.

10. A method of reducing the viscosity of a composition comprising a protein, the method comprising adding benzenesulfonic acid and camphorsulfonic acid as viscosity reducing excipients to a composition comprising at least one protein and glycine or proline.

11. The method according to claim 9, wherein: The protein is an antibody or an antigen-binding fragment thereof.

12. The method according to claim 9, wherein: The protein is included in a concentration of 10% to 25% (w / v).

13. The method according to claim 9, wherein: The glycine or protein is included at a concentration of 150 mM to 250 mM.

14. The method according to claim 9, wherein: The total concentration of the benzenesulfonic acid and the camphorsulfonic acid is 20 mg / mL to 50 mg / mL.

15. The method according to claim 9, wherein: The benzenesulfonic acid and the camphorsulfonic acid are included in a weight ratio of 1:0.5 to 1:2.5.

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