Art
[1]The present invention relates to liquid formulations of mumap otherwise Ah with anti -TNF alpha antibody, specifically.
BACKGROUND
[2]A tumor necrosis factor alpha (TNF alpha, TNFa) is a cytokine produced by a variety of cells such as mononuclear white blood cells and macrophages by stimulation, such as endotoxin. TNFa is activating the TNF receptor is the main mediator of the major inflammatory, immune, pathophysiological response that serves to elicit a response such as T- cell activation, thymocyte proliferation (Grell, M., et al. (1995) Cell , 83: 793-802).
[3]
Oh otherwise mumap it is a recombinant human immunoglobulin G1 monoclonal antibody to selectively bind to tumor necrosis factor alpha inhibit the immune response by tumor necrosis factor alpha in the body. Oh, unlike mumap was developed by the BASP Bioresearch Corporation, circa 1993, has been through a hard beotrae Laboratories Inc. (Abbott Laboratories) approved for sale in rheumatoid arthritis. Oh, unlike mumap are sold under the trade name of HUMIRA, HUMIRA is used in the treatment of Crohn's disease, ankylosing spondylitis, psoriatic arthritis, ulcerative colitis after receiving marketing approval in rheumatoid arthritis.
[4]
Oh, unlike mumap was developed by applying the phage display technology as the first fully human antibody drug developed an affinity enhancement has been causing mutations in the CDR. Oh otherwise mumap is called, has a molecular weight of about 148kD consisting of 1330 amino acids, also known as D2E7 (US Patent 6,090,382). Unlike ah mumap is a TNFa inhibitor and in combination with the p55 TNF, TNF and TNF receptor p75 in the cell surface to prevent reaction with each other to prevent a reaction to TNF the induction.
[5]
On the other hand, antibody drugs can lead to physical and chemical denaturation by a number of factors as a type of protein drugs. Denaturation of the protein loses the oxidation, deamidation, isomerization and chemical modification and which fragments may lead to structural modification such as animation or that this agglomeration, the pharmacological activity of the protein itself, if the protein is modified, such as, side effects in the body, as she may lead to unwanted immune responses. If the antibody fragment (Framentation), by changing the time staying in the binding affinity or the body can affect the pharmacological activity. There are also studies that the antibody fragments is induced aggregation of the antibody. In addition to airway reduces the pharmacological activity by aggregation. There are commercially available interferon-beta as compared to a product aggregate (aggregate) and a lot of study the content of the particles (particle) ratio higher product that neutralizing antibodies generated in vivo results (Barnard et al., 2013, J. Pharm. Sci . 102: 915). Saenggimyeon the neutralizing antibodies in the body even if injected protein drugs are neutralizing antibodies that bind to protein drugs to influence the stability, pharmacological effects, pharmacokinetics of the protein drug. Also in this denaturation and aggregation it was accentuated EpoetinAlfa out to be the cause of increased immunogenicity of Epoetin Alfa drugs. Therefore, for protein drugs not to lose its physiological activity during storage, and it is very important to manufacture the proper dosage form so that the protein is not Chemistry fragmentation, agglomeration, or particle. Therefore, it is going to actively research on the formulation of a number of protein drugs.
[6]
Detailed Description of the Invention
SUMMARY
[7]
Studies of the protein formulation has the purpose to be able to keep it stable until the patient is administered the optimal combination by properly mixing the various additives in consideration of the characteristics of each product. The primary purpose of the added additive is to control the physical properties of the stabilized mixture of the protein material. The additive is classified into surface active agent, stabilizers, preservatives, buffering agents and isotonic agents, and the like according to the purpose and properties. For antibody drugs have to find an effective therapeutic amounts of the protein is to be administered other than protein drugs. In addition, the high concentration formulation development is important because the route of administration, such as pain and difficulty of producing a large volume of patients hagieneun administered at a time when the subcutaneous injection. Increasing the concentration of the protein aggregates and high ground intermolecular interaction is increased, caused problems such as viscosity increase, gel Chemistry and precipitation of the solution. Of these, if the viscosity is excessively increased as well as the production it will not easily be difficult to administer to patients with increased injection pressure. Therefore, there is a predict the viscosity of the high concentration of the antibody solution to a number of lowering being studied.
Problem solving means
[8]
One object of the present invention is to provide liquid formulations of anti -TNF alpha antibody.
[9]
Another object of the present invention is to provide a method for preparing the liquid preparations.
[10]
Another object of the present invention uses a composition comprising a stabilizing agent, a surfactant, and arginine, to provide a method for increasing the stability of an anti -TNF alpha antibody.
[11]
Another object of the present invention does not include a buffering agent, by using a composition comprising a stabilizing agent, a surfactant, and arginine, to provide a method for increasing the stability of an anti -TNF alpha antibody.
Effects of the Invention
[12]
Wherein -TNFa antibody, liquid formulations of mumap otherwise specifically ah according to the present invention may allow for long-term storage by reducing the formation of by-products otherwise mumap Ah during the storage period. Further, it is possible to keep the long-term stability for the pharmacological efficacy of the contrast mumap O to prevent the denaturation and aggregation takes place in the production process and physical stress at the time of transportation. Therefore, the liquid preparation according to the present invention may be applied effectively to treat the field relevant to the pharmacological efficacy of the contrast mumap ah future.
Brief Description of the Drawings
[13]
Figure 1 shows the viscosity of the additive composition of the samples of Example 1. FIG.
[14]
Figure 2, illustrates an example embodiment 7, the number of sample and particles after the pump passes before the placebo sample.
Best Mode for Carrying Out the Invention
[15]
Hereinafter, the present invention will be described in more detail.
[16]
On the other hand, each described and embodiments of the disclosed herein may be applied to other embodiments and description of each. That is, any combination of the various elements described herein it is within the scope of the invention. In addition, it can not be said to the scope of the invention by the specific description will be described limits.
[17]
In addition, one of ordinary skill in the art may recognize or determine the number of equivalents for the particular embodiments of the invention described in this application using only routine experimentation. In addition, such equivalents are intended to be encompassed by the present invention.
[18]
One aspect of the present invention for solving the above problems is a liquid formulation of the anti -TNF alpha antibody.
[19]
[20]
As used herein, the term "anti -TNF alpha antibody" refers to antibodies that regulate the biological activity thereof binds to TNF alpha. More specifically, the antibodies may have the ability to bind to TNF-alpha inhibits the binding between TNF alpha and its receptor inhibit signal transduction by TNF alpha. In addition, such anti -TNF alpha antibody can be a monoclonal antibody.
[21]
The anti -TNF alpha antibody may be a form of an antibody fragment containing a full-length antibody or an antigen-binding site, are not particularly limited.
[22]
More specifically, the anti -TNF alpha antibody may be a recombinant human immunoglobulin G1 monoclonal antibodies, can mapil Oh no otherwise than specifically. Information about the Ah mumap Unlike those skilled in the art can be readily obtained from a known database.
[23]
The antibodies, but can be produced via recombinant DNA technology using mammalian cell expression system, it is not particularly limited.
[24]
The antibody may be contained in a therapeutically effective amount in the liquid formulations according to the invention. As specific examples, 1 to a 250mg / mL concentration, specifically, from 20 to 200mg / mL concentration, more specifically, 50mg / mL to about 200mg / mL of concentration, even more specifically, 50mg / mL, 100mg / mL, or but may be present at a concentration of 130mg / mL, it is not particularly limited.
[25]
[26]
Liquid formulations of the present invention may include stabilizers, surfactants, and arginine in addition to anti -TNF alpha antibody. The liquid formulation may be a solution formulation that can store -TNF alpha antibody stably wherein.
[27]
Specifically, there is a well-known protein stability assays in the field of the art can be used to measure the stability of an anti -TNF alpha antibody. Stability can be measured at a selected temperature during the selected time. For rapid testing, the formulation is more, for higher or "accelerated" temperature, for example, can be stored at 40 ℃ for at least 2 weeks to 1 month, at this point of time-dependent stability is measured.
[28]
Means that in the present invention, "anti -TNF that it granted to stabilize the alpha antibody" is a period of time for a particular storage conditions, in particular under a certain amount of loss of the active ingredient under certain temperature, such as less than 10%. Usually, at 5 ± 3 ℃ 2 nyeon, at 25 ± 2 ℃ 6 months or at 40 ± 2 ℃ wherein for one month to about two months -TNF alpha antibody is at least 90%, specifically, keeping the residual rate of at least about 92% such agents if may be understood to be stable.
[29]
[30]
A stabilizer contained in the liquid formulations of the present invention may be a polyol, an amino acid, or a combination thereof. Here, the amino acid may be a different amino acid other than arginine.
[31]
Specifically, the stabilizer are 1) one kind of the polyol, and 2) a combination between the kind of polyol and one kind of amino acid, and 3) one kind of the polyol, the first amino acid, and a combination of the second amino acid, and 4) first a combination of a polyol and a second polyol, and 5) a first polyol, a second polyol and a type of a combination between an amino acid, 6) a first polyol, a second polyol, the first amino acid, and a combination between the two amino acids, or 7) It can be a kind of amino acid.
[32]
More specifically, the polyol may be any of mannitol, sucrose, trehalose, PEG, or combinations thereof, may be more specifically, sucrose, trehalose, PEG, or a combination thereof. The PEG may be a specifically PEG400 or PEG4000, is not particularly limited. In the preparation polyols may be present in a concentration of 0.1 to 100 mg / mL.
[33]
[34]
More specifically, the different amino acid other than the arginine may be glycine, leucine, isoleucine, phenylalanine, or proline. In the formulation is an amino acid may be present in a concentration of 1 to 300 mM.
[35]
In addition, the present invention is in the term "amino acid" also includes substantially all the analogues, solvate, hydrate, stereoisomer, and a pharmaceutically acceptable salts thereof of the amino acid representing the same effect.
[36]
In the present invention, including the salts derived from the term "pharmaceutically acceptable salt" is a pharmaceutically acceptable mineral acid is organic acid, or a base. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, glycolic acid, lactic acid, salicylic acid, succinic acid, -p- toluene sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid , there may be mentioned benzoic, malonic, naphthalene-2-sulfonic acid, benzene sulfonic acid. Salts derived from an appropriate base may include an alkali earth metal, and ammonium, such as an alkali metal, such as magnesium, sodium, potassium.
[37]
In addition, the term "solvate" used in the present invention refers to a amino acid or a salt thereof to form a complex with the solvent molecules.
[38]
[39]
Still more specifically,
[40]
The stabilizing agent (i) sucrose or trehalose, (ii) number of PEG, (iii) an average molecular weight of 200 to 600 PEG, or a number average molecular weight of from 1000 to 8000 glycine or leucine, and (iv) wherein (i) - (iii) of, but may be selected from the group consisting of two or more thereof, is not particularly limited.
[41]
As a more specific example, the stabilizers are: 1) sucrose, either of trehalose, and PEG400, 2) sucrose or a combination of trehalose and glycine or leucine, 3) sucrose or a combination of trehalose and glycine and leucine, 4) sucrose or trehalose and PEG4000 combination of five) sucrose or a combination of trehalose, PEG4000, and glycine, 6) sucrose or a combination of trehalose, PEG4000, and leucine, 7) sucrose or a combination of trehalose, PEG4000, glycine, and leucine, and 8) glycine It may be a one selected from the group consisting of, but is not particularly limited.
[42]
[43]
Surface active agent contained in the liquid formulations of the invention may be a non-ionic surfactant. More specifically, the surfactant can meoil polysorbate or Pollock used.
[44]
Specifically, the surfactant is polysorbate 80, polysorbate 20 or poloxamer 188, but can work, is not particularly limited.
[45]
In the formulation the surfactant may be present in a concentration of 0.1 to 5 mg / mL.
[46]
[47]
Arginine it contained in the liquid formulations of the present invention may exist in salt form. More specifically, it may be a pharmaceutically acceptable salt form.
[48]
More specifically, the arginine may be an arginine hydrochloride (Arginine hydrochloride) form, it is not particularly limited.
[49]
In the formulation arginine may be present in a concentration of from 0.1 to 200 mM. More particularly, the Arginine if said antibody is present in the formulation as a 100mg / mL There may be present in a concentration of 0.1 to 140mM, if the antibody is present in the formulation as a 50mg / mL can be present at a concentration of 0.1 to 100mM, but It is not particularly limited.
[50]
The arginine can be included in the liquid formulations of the present invention as a viscosity depressant.
[51]
By including arginine, liquid formulations of the invention, but may have a viscosity of from about 1 to about 6 cps, it is not particularly limited. Measurement of the viscosity can take place using a variety of methods known in the art, for example, but can be carried out in the same manner as described herein in Example 1, it is not particularly limited.
[52]
[53]
Liquid formulations of the present invention may further comprise an antioxidant.
[54]
As used herein, the term "antioxidants" may serve to suppress the impurity generation that can occur by the oxidation reaction or the like of the protein in solution.
[55]
As such an antioxidant, sodium bisulfate, ascorbic acid, ascorbyl palmitate, citric acid, butyl hydroxy anisole (BHA), butylhydroxytoluene (BHT), thioglycerol, propyl gallate, methionine, sodium ascorbate, It may be mentioned sodium citrate, sodium sulfide, sodium sulfite, EDTA, and other antioxidants. However, without being limited thereto.
[56]
Antioxidant to the, in particular methionine in the formulation but be present in a concentration of 1 to 50 mM, it is not particularly limited.
[57]
[58]
In addition, pH of the liquid preparation of the present invention, but from 4 to 6, it is not particularly limited.
[59]
[60]
On the other hand, not particularly limited, the liquid formulation may be one which does not contain additional salt and / or buffer.
[61]
As a non-limiting example, the formulation containing 50mg / mL or more anti -TNF alpha antibody may not comprise additional salts and / or buffer.
[62]
As noted in the embodiment of the present invention, the preparation according to the present invention it does not include these in comparison to the formulation containing salt, buffer, or both may be given a higher stability to heat the anti -TNF alpha antibody. However, it is not particularly limited.
[63]
On the other hand, the high concentration of the antibody drugs, for example 50mg / to the formulation of drugs which have mL or more antibodies present additional buffering agents, or the solution of the osmotic pressure if the amount of further additives is not used, causing a departure from the osmotic pressure range is similar to the body fluid decreases when administered pain there is comfort of the patient can be increased.
[64]
[65]
On the other hand, not particularly limited to, a liquid formulation according to the present invention may have the following effects:
[66]
Liquid preparation according to the invention containing arginine is the aggregation of anti -TNF alpha antibody protein is inhibited as compared to formulations that do not contain ahreugininreul may indicate the content of the relatively low molecular products (HMW) / or, not include arginine as compared to formulations that are to suppress the generation of the isomeric acid antibody may comprise a relatively low acid isomeric antibody. In particular, the liquid formulations according to the invention can be generated to hold the reduction in the antibody, and / or the effect of agglomeration and particle generation reduced to a particular physical stress due to degeneration.
[67]
[68]
Solution formulation according to the present invention may include a preservative in addition. The preservative is in a, such preservatives to mean a compound added to a pharmaceutical formulation benzalkonium chloride, benzethonium benzamide to act as an antibacterial agent, when a Dean-chloro-hexyl, phenol, m- cresol, benzyl alcohol, methyl paraben, propyl paraben, chlorobutanol, o- cresol, can be used in the p- cresol, chloro-cresol, phenyl mercury nitrate acid (phenylmercuric nitrate), thimerosal, benzoic acid, etc., without being limited thereto. These preservatives may be used in used alone or optionally in combination of two or more.
[69]
[70]
Liquid preparation according to the invention may be in the form of a pharmaceutical composition.
[71]
Further, in addition to the above-mentioned components may include a variety of pharmaceutically acceptable carriers.
[72]
Formulations of the present invention is rheumatoid arthritis, psoriasis, psoriatic arthritis, layering spinal arthritis (such as ankylosing spondylitis, radiological as ankylosing spondylitis severe layering spinal arthritis is not OK), vasculitis, Alzheimer's disease, ulcerative colitis, Behcet's colitis, purulent sweat gland salt, uveitis, juvenile idiopathic arthritis, juvenile plaque psoriasis, or Crohn's disease (Crohn's disease including adult, pediatric Crohn's disease) may be used in the prevention or treatment, such as, without being limited thereto.
[73]
The formulations according to the present invention is not within the oral administration, or subcutaneous, intramuscular, intraperitoneal, intrasternal, transdermal, and intravenous injection, and but can be administered in vivo by parenteral administration, including injection, limited.
[74]
[75]
Yet another aspect of the invention is a method for producing the liquid preparation of the anti -TNF alpha antibody, comprising the step of mixing of the anti -TNF alpha antibody, stabilizers, surfactants, and arginine.
[76]
It is as described above for these terms. In addition, applied both to the specific examples of the present embodiment, each term is obvious.
[77]
[78]
Further, another aspect of the present invention is a method of using a composition comprising a stabilizing agent, a surfactant, and arginine, increase the stabilization of the anti -TNF alpha antibody.
[79]
It is as described above for these terms. In addition, applied both to the specific examples of the present embodiment, each term is obvious.
[80]
[81]
Another object of the present invention does not include a buffering agent, by using a composition comprising a stabilizing agent, a surfactant, and arginine, to provide a method for increasing the stability of an anti -TNF alpha antibody.
[82]
It is as described above for these terms. In addition, applied both to the specific examples of the present embodiment, each term is obvious.
Mode for the Invention
[83]
Will now be described in more detail through the practice of the present invention. However, these examples are intended to illustrate the invention by way of example, it is not the scope of the present invention is limited to these Examples.
[84]
[85]
< Example 1>
[86]
According to the additive O mumap otherwise make the viscosity-lowering effect of the solution and stability
[87]
[88]
Oh the mumap liquid formulation to ensure an additive for use in the preparation of sucrose 55mg / mL, Methionine 5 mM, polysorbate 80 1 mg / mL, Oh otherwise mumap 100 mg / mL, pH Formulation 1 5.2 The composition was prepared otherwise .
[89]
Further more in arginine hydrochloride, lysine hydrochloride, leucine, isoleucine, phenylalanine, glutamic acid, glycine, proline, alanine, sodium chloride, calcium chloride, Rheosense's mVROC device by respectively the addition of magnesium chloride to prepare a formulation 2 to 13 with the composition of Formulation 1 used to measure the viscosity. To the type and the concentration and viscosity of each formulation of the additive added to the formulation shown in Table 1 and Figure 1.
[90]
TABLE 1
Formulation # Additive composition Viscosity (cp)
Formulation 1 - 3.23
Formulation 2 20 mM arginine hydrochloride 3.04
Formulation 3 40 mM lysine hydrochloride 3.14
Formulation 4 Leucine 40 mM 3.28
Formulation 5 40 mM isoleucine 3.26
Formulation 6 Phenylalanine 15 mM 3.21
Formulation 7 7.5 mM glutamate 3.07
Formulation 8 40 mM glycine 3.20
Formulation 9 40 mM proline 3.20
Formulation 10 Alanine 40 mM 3.21
Formulation 11 40 mM NaCl 3.12
Formulation 12 20 mM calcium chloride 2.94
Formulation 13 Magnesium chloride, 20 mM 3.01

[91]
[92]
When the viscosity values ​​of the respective formulations shown in Table 1 for sucrose, methionine, polysorbate 80, and ah Formulation 1 consisting of mumap different viscosity was 3.23. As compared the case of the addition of amino acids leucine, isoleucine, phenylalanine, glycine, proline, such as formulation viscosity, the viscosity was not significantly different to 3.20 ~ 3.28. On the other hand, if the addition of arginine hydrochloride, lysine hydrochloride, glutamic acid, sodium chloride, calcium chloride, magnesium chloride, etc. The viscosity was found to decrease the viscosity to 2.94 ~ 3.14.
[93]
[94]
Each formulation was sealed sterilizing by filtration after each containing 0.4 mL to 1.0 mL capacity glass syringe for about 40 to a pore size of 0.2 ㎛ filter in order to compare the stability of each formulation o were kept two months C. After storage and analyzed the content of the polymer product (High molecular weight impurities, HMW) and O otherwise fragment the low molecular weight product of mumap molecule (Low molecular weight impurities, LMW) and the like of various samples to analyzed by SE-HPLC and oligomers, aggregates. First, the viscosity drop formulations, i.e. formulations 2, Formulation 3, Formulation 7 Formulation 11, Formulation 12, Formulation 13 and exhibited a SE-HPLC results of formulations 1, the control group in Table 2 below. In addition, formulations that are not significantly different viscosity, i.e. formulations 4, Formulation 5, Formulation 6, Formulation 8, Formulation 9, Formulation 10 and shows the SE-HPLC results of formulations 1, the control group are shown in Table 3.
[95]
TABLE 2
Keep ago 40 O C 2 months of storage
HMW (%) LMW (%) Total (%) HMW (%) LMW (%) Total (%)
Formulation 1 0.33 0.54 0.87 1.46 5.13 6.59
Formulation 2 0.33 0.53 0.86 1.31 5.21 6.52
Formulation 3 0.33 0.53 0.85 1.42 5.18 6.61
Formulation 7 0.33 0.56 0.89 1.40 5.06 6.45
Formulation 11 0.35 0.55 0.89 1.70 5.40 7.10
Formulation 12 0.33 0.54 0.87 1.36 6.02 7.38
Formulation 13 0.34 0.54 0.88 1.72 5.70 7.42

[96]
TABLE 3
Keep ago 40 O C 2 months of storage
HMW (%) LMW (%) Total (%) HMW (%) LMW (%) Total (%)
Formulation 1 0.33 0.54 0.87 1.46 5.13 6.59
Formulation 4 0.33 0.52 0.85 1.27 4.81 6.07
Formulation 5 0.33 0.56 0.88 1.28 4.74 6.02
Formulation 6 0.34 0.52 0.86 1.32 4.85 6.17
Formulation 8 0.33 0.48 0.81 1.21 4.77 5.98
Formulation 9 0.32 0.55 0.87 1.14 4.80 5.94
Formulation 10 0.34 0.56 0.90 1.24 4.84 6.08

[97]
[98]
Table 2 The results of storage but prior to HMW, LMW and similar amount of total impurities, 40 o For the formulation total impurities, including sodium chloride, after 2 months storage at C 11 for 7.10%, the formulation 12 contains calcium chloride in the case of 7.38%, dosage form 13 including the magnesium chloride was found to 7.42%, up to larger in case of total impurities 6.59% of the formulation 1 control group. While arginine hydrochloride, lysine hydrochloride, formulations containing the glutamic acid 2, Formulation 3, in the case of formulations 7 40 o the total impurities 6.59% and the impurity level of C in the control group as two months of storage is 6.45 ~ 6.61% total amount of impurities Formulation 1 It was maintained
[99]
The results of Table 3, leucine, isoleucine, phenylalanine-glycine, in the case of the formulation by the addition of amino acid such as proline 40 o C 2 gaewol impurity content of the formulation 1 of total impurity amount of the control group to 5.94 ~ 6.17% after storage 6.59 in contrast to the% it confirmed that significantly reduced. Thus it was confirmed that the amino acid leucine, isoleucine, phenylalanine, glycine, proline, etc. Oh contributing to the stability of the otherwise mumap.
[100]
[101]
< Example 2>
[102]
Stability assessment of the amount of arginine hydrochloride 1
[103]
[104]
In order to evaluate the stability of the O Unlike mumap formulations according to the content of arginine hydrochloride was prepared in a formulation as follows:
[105]
Sucrose, 55mg / mL, methionine 5mM, polysorbate 80 1mg / mL and O otherwise were prepared formulation 14 such that the mumap 100mg / mL, the composition by the addition of arginine hydrochloride 20mM and arginine hydrochloride 40mM the composition of Formulation 14 Formulation 15 and producing a dosage form 16, it was charged by the 1mL 0.4mL glass syringe. Each syringe 40 o After 2 months storage at C were evaluated for stability by analyzing a SE-HPLC. HMW content of the composition and storage before / after each of the formulations shown in Table 4.
[106]
TABLE 4
Formulation Furtherance HMW (%)
40 O C Storage 2 months ago 40 O C 2 months of storage
Formulation 14 Sucrose, 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, arginine hydrochloride 0mM 0.17 1.00
Formulation 15 Sucrose, 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, 20mM arginine hydrochloride 0.17 0.82
Formulation 16 Sucrose, 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, 40mM arginine hydrochloride 0.16 0.82

[107]
[108]
In the SE-HPLC results of Table 4, HMW content of the sample before storage when the content of arginine hydrochloride hayeoteul increase in 0mM (formulation 14) to 20mM (formulation 15) and 40mM (formulation 16), all in 0.16 ~ 0.17% were similar, 40 o C 2 gaewol storage after HMW content when the formulation 14 does not contain the arginine hydrochloride 1.00%, the amount of arginine hydrochloride 20mM and 40mM formulations 15, for a 16-arginine hydrochloride to 0.82% is 20mM or 40mM if they contain about HMW generated was reduced than in the case that do not contain arginine hydrochloride. Therefore, arginine hydrochloride was confirmed that the anti-aggregation effects of different ah mumap.
[109]
[110]

[111]
Charge isomeric antibody according to the arginine content (charge variant) produced Comparison of
[112]
[113]
Creating a formulation that does not contain the arginine formulation comprising 40 to compare the charge isomeric antibodies degree according to the arginine content o compared the C 1 month after the charge to the CEX-HPLC isomeric antibodies aspect stored at. It is shown for each formulation and storage before and after the charge-isomeric content of antibodies in Table 5 below.
[114]
Table 5
Formulation Formulations Summary 40 O C 1 Keep months ago 40 O C 1 months of storage
Acidic K0 K1 Basic Acidic K0 K1 Basic
A Sucrose, 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, Oh otherwise mumap 100mg / mL 13.36 70.77 12.44 3.44 33.39 51.58 10.70 4.33
B Sucrose, 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, Oh otherwise mumap 100mg / mL, 20mM arginine hydrochloride 13.15 71.14 12.41 3.30 31.31 52.51 11.16 5.01

[115]
Formulation A is sucrose 55mg / mL, methionine 5mM, polysorbate 80 (PS80) 1mg / mL and O otherwise mumap 100mg / mL were prepared to include the formulation B is arginine hydrochloride (Arginine Hydrochloride) the composition of Formulation A for It was prepared to include an additional 20mM. 40 o C 1 gaewol storage acid isomeric antibody content was similar been, stored acidic isomeric Comparing the antibody content of the case of formulation B containing arginine hydrochloride formulations A content of the prepared low content of acidic isomeric antibody, K0 after before the it can be seen more.
[116]
[117]
Therefore, unlike the arginine hydrochloride ah it found that the lower the acidity of the isomeric antibody mumap.
[118]
[119]

[120]
Type formulation and comparative formulation stability of the surface active agent
[121]
[122]
Unlike ah according to the kind of the surface active agent was prepared in the following formulation of the same composition in order to compare the stability of the liquid formulations mumap.
[123]
Example 2 The formulation was prepared consisting of 17 such that the sucrose 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL and O otherwise mumap 100mg / mL, such as in the formulation 14. In addition, the content of the surfactant is prepared in the same manner replacing the kind of the surface active agent as polysorbate 20 and poloxamer 188 formulation 18 and formulation 19 and fixed.
[124]
And filtering each of the filtered formulation is filled by a 1mL 0.4mL glass syringe 40 o was one months storage in C. The impurity content of the HMW and LMW samples were analyzed before and after storage using a SE-HPLC. Configuration and 40 of dosage form 17 ~ 19 o The results of analyzing the content of C SE-HPLC before and after 1 months storage are shown in Table 6.
[125]
TABLE 6
40 O C 1 Keep months ago 40 O C 1 months of storage
HMW (%) LMW (%) Total (%) HMW (%) LMW (%) Total (%)
Formulation 17 Sucrose, 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, pH5.2 0.14 0.37 0.51 0.46 2.05 2.52
Formulation 18 Sucrose, 55mg / mL, 5mM methionine, polysorbate 20 1mg / mL, pH5.2 0.15 0.38 0.53 0.45 2.04 2.49
Formulation 19 Sucrose, 55mg / mL, 5mM methionine, Poloxamer 188 1mg / mL, pH5.2 0.15 0.37 0.53 0.44 1.99 2.43

[126]
The results of the table 640 in accordance with the type of surfactant o C one month storage HMW and LMW content before and after the not much different. That is, poly sorbitan the formulation and stability of the formulation containing poloxamer 188, including the formulation, polysorbate 20, including the polysorbate 80 were similar to each other.
[127]
Therefore, polysorbate 80, polysorbate 20 and the poly poloxamer 188 was confirmed that the influence on the stability of the otherwise similar to each other mumap Ah.
[128]
[129]
< Example 5>
[130]
Comparison type formulation and the stability of the polyol
[131]
[132]
Unlike ah according to the kind of the polyol was prepared in the following formulation of the same composition in order to compare the stability of the liquid formulations mumap.
[133]
Example 2 The formulation 20 is configured such that the sucrose 55mg / mL, methionine 5mM, polysorbate 80 1mg / mL and O otherwise mumap 100mg / mL, such as Formulation 14 were prepared (the same as in the formulation 17 in Example 4). In addition, the total amount of polyol was prepared with the same fixed trehalose the type of polyol, formulations 21-23 replaced with PEG400 and PEG4000. And filtering each of the filtered formulation is filled by a 1mL 0.4mL glass syringe 40 o was one months storage in C. The impurity content of the HMW and LMW samples were analyzed before and after storage using a SE-HPLC.
[134]
Configuration and 40 of dosage form 20 ~ 23 o The results of analyzing the content of C SE-HPLC before and after 1 months storage are shown in Table 7.
[135]
Table 7
40 O C 1 Keep months ago 40 O C 1 months of storage
HMW (%) LMW (%) Total (%) HMW (%) LMW (%) Total (%)
Formulation 20 Sucrose, 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, pH5.2 0.14 0.37 0.51 0.46 2.05 2.52
Formulation 21 Trehalose 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, pH5.2 0.14 0.36 0.50 0.46 2.03 2.49
Formulation 22 PEG400 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, pH5.2 0.16 0.38 0.54 0.62 2.02 2.63
Formulation 23 PEG4000 55mg / mL, 5mM methionine, polysorbate 80 1mg / mL, pH5.2 0.17 0.37 0.53 0.85 2.05 2.90

[136]
The results of the table 740 in accordance with the type of the polyol o has a C 1 gaewol keep HMW and LMW content before and after the confirmed that differs greatly. 40 for all formulations o Although C 1 gaewol storage before HMW and LMW impurity content is similar, 40 o C 1 gaewol storage after as compared to formulations 23, including the case of the formulations 20-22 is HMW content, including sucrose and trehalose, and PEG400 PEG4000 relatively low. LMW content was similar to both. Therefore, according to the type of polyol used it was confirmed that there may be differences in the formulation between stability among them was confirmed that the sucrose and trehalose has an effect to improve stability than other polyols.
[137]
[138]
< Example 6>
[139]
Polyols, arginine, methionine, the surfactant, and further comprising a stabilizer formulation and Humira stability compared with formulations
[140]
[141]
And by changing the methionine 5mM different surface active agent in the composition used as a stabilizer and the polyol type, adding a stabilizer or absence and arginine hydrochloride content, etc. to prepare a sample by constructing a variety of compositions and manufacturing compositions in commercial Humira formulation 40 o and then stored at C analyzed by SE-HPLC was compared to the stability of each formulation. The composition of each formulation in Table 8, 40 of each composition o C 2 gaewol impurity content before and after storage are shown in Table 9.
[142]
Table 8
Formulation # 　Formulation Composition
24 Unlike mumap Oh 100 mg / mL, 5mM methionine, arginine hydrochloride 0mM, Polysorbate 80 1mg / mL, sucrose 55mg / mL
25 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 20mM, polysorbate 80 1mg / mL, sucrose, 55mg / mL
26 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, polysorbate 80 1mg / mL, sucrose, 55mg / mL
27 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose, 55mg / mL
28 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose, 55mg / mL, 20mM glycine
29 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose, 55mg / mL, 20mM leucine
30 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose, 55mg / mL, 10mM glycine, 10mM leucine
31 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 55mg / mL
32 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 55mg / mL, 20mM glycine
33 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 55mg / mL, 20mM leucine
34 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 55mg / mL, 10mM glycine, 10mM leucine
35 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose 27.5mg / mL, PEG4000 27.5mg / mL
36 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose 27.5mg / mL, PEG4000 27.5mg / mL, 20mM glycine
37 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose 27.5mg / mL, PEG4000 27.5mg / mL, 20mM leucine
38 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, sucrose 27.5mg / mL, PEG4000 27.5mg / mL, 10mM glycine, 10mM leucine
39 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 27.5mg / mL, PEG4000 27.5mg / mL
40 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 27.5mg / mL, PEG4000 27.5mg / mL, 20mM glycine
41 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 27.5mg / mL, PEG4000 27.5mg / mL, 20mM leucine
42 Oh otherwise mumap 100 mg / mL, 5mM methionine, arginine hydrochloride 40mM, Poloxamer 188 1mg / mL, trehalose 27.5mg / mL, PEG4000 27.5mg / mL, 10mM glycine, 10mM leucine
43 Oh otherwise mumap 100mg / mL, commercial formulation HUMIRA *
44 Oh otherwise mumap 50 mg / mL, 5mM methionine, arginine hydrochloride 40mM, polysorbate 80 1mg / mL, sucrose, 55mg / mL
45 Oh otherwise mumap 50mg / mL, commercial formulation Humira *

[143]
*상용 휴미라 제형: Sodium Phosphate Monobasic Dihydrate (0.86　mg/mL), Sodium Phosphate Dibasic Dihydrate (1.53　mg/mL), Sodium Citrate (0.3　mg/mL), Citric Acid Monohydrate (1.3　mg/mL), Mannitol (12　mg/mL), Sodium Chloride (6.16　mg/mL), Polysorbate 80 (PS80) (1　mg/mL), pH　5.2
[144]
[145]
Table 9
Formulation # 40 O C Storage 2 months ago 40 O C 2 months of storage
HMW(%) LMW (%) Total(%) HMW(%) LMW (%) Total(%)
24 0.17 0.44 0.60 1.00 4.42 5.43
25 0.17 0.45 0.62 0.82 4.49 5.32
26 0.16 0.41 0.57 0.82 4.69 5.52
27 0.17 0.43 0.60 0.85 4.57 5.42
28 0.17 0.45 0.61 0.77 4.55 5.32
29 0.16 0.44 0.60 0.80 4.55 5.34
30 0.17 0.44 0.60 0.79 4.54 5.33
31 0.17 0.45 0.62 0.76 4.58 5.34
32 0.17 0.45 0.62 0.73 4.55 5.28
33 0.17 0.43 0.60 0.76 4.55 5.31
34 0.16 0.44 0.60 0.76 4.57 5.33
35 0.18 0.45 0.62 0.99 4.56 5.55
36 0.17 0.44 0.61 1.06 4.50 5.56
37 0.17 0.43 0.61 1.02 4.50 5.52
38 0.18 0.44 0.62 0.95 4.47 5.42
39 0.17 0.43 0.61 0.96 4.52 5.48
40 0.17 0.44 0.62 0.97 4.51 5.48
41 0.17 0.42 0.59 0.99 4.50 5.49
42 0.18 0.43 0.61 0.99 4.55 5.55
43 0.29 0.44 0.73 1.44 5.52 6.95
44 0.19 0.43 0.62 0.50 4.62 5.12
45 0.29 0.45 0.74 1.15 5.66 6.81

[146]
In Table 8, the combination of sucrose and trehalose, and combinations and trehalose and sucrose and of PEG4000 PEG4000 was used as a polyol, a surface active agent include a polysorbate 80 and poloxamer 188 were used. Arginine is to determine the role of further additives, has been used is 0, 20 or 40mM glycine (Gly), leucine (Leu) or to design the formulation of a combination of geulrisinwa leucine Oh otherwise HUMIRA stability at mumap 100mg / mL commercial It was compared with the formulation. Stability at 50mg / mL Figure (in contrast with the formulations Ah 26 except the content of the same mumap) HUMIRA commercial formulation and formulation 44 was compared to the composition of the.
[147]
Comparing the stability of each formulation in Table 9, it was confirmed that all of the formulation indicates a superior stability than Humira commercial formulation. Total impurities before storage was similar to all 0.57 to 0.74. However, 40 o C In the stability after 2 months storage, Oh otherwise the content of mumap of 100mg / mL of HUMIRA commercial formulation (formulation 43) and O otherwise mumap content of 50mg / mL of HUMIRA commercial formulation (formulation 45) compared to other formulations Oh Unlike confirmed that more mumap low impurity content.
[148]
Therefore, the various polyols and a surfactant, the formulation consisting of arginine hydrochloride, and zero additional stabilization was confirmed to be a more stable formulation in terms of increased impurity than the commercial Humira formulation.
[149]
[150]
Also 40 than the formulation with the use of sucrose, trehalose, the PEG 4000 with a polyol in the preceding embodiment 5 o compared to the total impurity content when C 1 gaewol kept about 0.4% was high, in the present embodiment, though two months yieoteumedo storage time using a mix of sucrose or trehalose and PEG4000 when Ah was compared to otherwise similar to the impurity content of mumap exclusive use of sucrose and trehalose. As a result, it was confirmed to be replaced with sucrose or part of a larger molecular weight PEG of trehalose can be used as a way to maintain the osmotic pressure and stability when using an additional additive in order to obtain an additional effect, such as antioxidants.
[151]
[152]

[153]
Commercial Humira stability compared evaluation of the formulation arginine and mechanical stresses (mechanical stress) of the hydrochloride formulation including
[154]
[155]
In order to compare the stability of the mechanical stress of the formulation containing the commercial Humira formulation with arginine hydrochloride, create a sample of the following composition was evaluated in the same pump can pass before and after the particles passed through the rotary piston pump (rotary piston pump).
[156]
Also evaluated the Oh otherwise only placebo (placebo) In the same condition before and after the water pump passes through the particles was passed through a rotary-piston pump in creating mumap except in the composition of each sample to ensure that the particle measured ah mumap otherwise derived. A method of assessing the number of particles was used the Protein Simple's micro flow imaging device. The composition of each sample is shown in Table 10, showing the number of placebo pump passed before and after the particles of each sample and the samples in Table 11 and FIG.
[157]
[Table 10]
Formulation 46 Oh otherwise mumap 100 mg / mL, sucrose 55 mg / mL, 40 mM arginine hydrochloride, methionine, 5 mM, polysorbate 80 1 mg / mL, pH5.2
Formulation 47 Oh otherwise mumap 50 mg / mL, sucrose 55 mg / mL, 40 mM arginine hydrochloride, methionine, 5 mM, polysorbate 80 1 mg / mL, pH5.2
Formulation 48 Oh otherwise mumap 50 mg / mL, phosphate is sodium hydrate (Sodium phosphate monobasic dihydrate) 0.86 mg / mL, a sodium hydrogen phosphate hydrate (Sodium phosphate dibasic dihydrate) 1.53 mg / mL, sodium citrate (Sodium citrate) 0.3 mg / mL, citric acid monohydrate (citric acid monohydrate), 1.3 mg / mL, mannitol 12 mg / mL, NaCl 6.16 mg / mL, polysorbate 80 (PS80) 1 mg / mL, pH5.2 (composition commercial Humira)

[158]
[Table 11]
The particle concentration (# / mL)
placebo Samples (ah, unlike included mumap)
Before passing through the pump After passing through the pump Before passing through the pump After passing through the pump
Formulation 46 2667 3618 699 85743
Formulation 47 2667 3618 769 53734
Formulation 48 1213 7938 1917 150617

[159]
Formulation 46 and formulation 47, as shown in Table 10 is the same as the composition of the additive, and, unlike the O content of 46 mumap formulation is 100mg / mL, dosage form 47 is different from a 50mg / mL. For formulation 48 was the same composition and O otherwise control the mumap content and commercial Humira. Results for each formulation was analyzed the number of particles before and after passed through a rotary piston pump, a table 11 and formulation 46, such as 2 85 743 pieces / mL, dosage form 47 is a high dosage form content of mumap otherwise ah, with 53 734 pieces / mL 46 is different Oh it was confirmed that the content of the particles is further increased mumap lower dosage form 47. Whereas commercial Humira was the number of particles after passing through the pump case of the formulation of 48 dosage form highly dog ​​150 617 / mL formulated in 46 and 47 contrast. After also passing through the particle concentration for all formulations placebo pump 3618 / mL and 7938 / contrast Ah in mL after pump passage of the specimen containing a mumap significantly less than the particle concentration, after the pump passage ah otherwise measured at mumap containing sample Oh, it was confirmed that the particles, unlike mumap origin. Accordingly, there is a dosage form containing arginine hydrochloride can be seen that the Oh otherwise mumap be protected against mechanical stresses than commercial Humira formulation.
[160]
[161]
< Example 8>
[162]
Arginine hydrochloride concentration by viscosity lowering effect test
[163]
[164]
Oh otherwise the viscosity of the addition of arginine to the solution mumap to determine the arginine concentration range of the drop occurring the experiment was as follows. Arginine hydrochloride (ArgHCl) as a control that does not contain, Oh otherwise prepare a sample to include mumap 100mg / mL and polysorbate 80 1mg / mL, or Ah otherwise mumap 50mg / mL and 80 1mg / mL polysorbate. As test groups, gamyeo increased by a 20mM concentration of arginine hydrochloride on the composition of each of the control group the arginine hydrochloride to prepare a sample containing a final concentration of 180mM maximum. The pH of all samples was about 5.2. The viscosity of the prepared sample was measured using a Rheosense's mVROC device. Viscosity The measurement results are shown in Table 12.
[165]
[Table 12]
ArgHCl(mM) 100mg / mL Oh unlike mumap (Unit: cp) 50mg / mL Oh unlike mumap (Unit: cp)
0 2.71 1.47
20 2.59 1.42
40 2.59 1.42
60 2.62 1.42
80 2.62 1.45
100 2.63 1.48
120 2.67 1.49
140 2.70 1.50
160 2.75 1.52
180 2.79 1.54

[166]
Table 12 Ah otherwise mumap 100mg / mL, polysorbate 80 1mg / mL, and looking at the arginine hydrochloride (ArgHCl) the viscosity of the additive formulation at various concentrations, Oh otherwise mumap 100mg / ml, which does not include ArgHCl, polysorbate 80 1mg / mL viscosity of the solution was 2.71 cp. When the ArgHCl to this was added to a final concentration of 20 ~ 120mM it can be confirmed that the viscosity of the composition prepared without addition of the ArgHCl dropping. If ArgHCl are added to a final concentration of 140mM it did not have the addition of ArgHCl composition and viscosity was similar. If ArgHCl are added to a final concentration of at least 160mM there did not receive the effect of lowering the viscosity, ArgHCl confirmed that the non-viscosity is higher than addition of the formulation. Oh contrast to mumap 50mg / mL and polysorbate 80 1mg / in the case of the formulation including mL, was the viscosity of the case which does not include ArgHCl 1.47 cp, is 1.42 ~ 1.45 viscosity when the ArgHCl concentration of 20 ~ 80 mM ArgHCl a viscosity lower than the US added to the formulation. The viscosity of the addition of ArgHCl to a final concentration of 100mM was an increase in viscosity than when ArgHCl this was similar to the non-addition of the viscosity of the composition, without addition of the addition the ArgHCl to a final concentration of at least 120mM.
[167]
Therefore, it was confirmed that 100mg / mL O otherwise mumap added to ArgHCl of 140mM or less it is possible to lower the viscosity for the solution, 50mg / mL O contrast to lower the viscosity for mumap solution is added arginine in 100mM or less.
[168]
[169]
< Example 9>
[170]
Oh otherwise mumap confirmation buffer and salt effects on the formulation Test 1
[171]
[172]
Oh contrast was prepared a sample of the formulation did not include a buffer and a salt to determine the buffer and salt effects on the stability of mumap, to thereby prepare a sample of the addition of a buffer or salt formulated in a dosage form 40 o 2 weeks in C and 1 month storage was compared to the stability by SE-HPLC, to measure the pH of each sample.
[173]
[Table 13]
Formulation # Formulation Add buffer and Salt zerotime 40 O C 2 weeks of storage 40 O C 1 months of storage
HMW LMW Total HMW LMW Total HMW LMW Total
A-1 Oh, unlike mumap 100mg / mL, PS801mg / mL, pH 5.2 no buffer/no salt 0.36 0.38 0.74 0.47 1.19 1.66 0.65 2.04 2.69
A-2 Sodium phosphate / sodium citrate (Humira Composition Buffer) * 0.38 0.39 0.76 0.59 1.25 1.84 0.92 2.27 3.19
A-3 NaCl 100mM 0.37 0.41 0.78 0.66 1.42 2.09 0.86 2.50 3.35
A-4 Ammonium sulfate (Ammonium sulfate) 100mM 0.37 0.41 0.78 0.50 1.42 1.92 0.69 2.54 3.24
A-5 Sodium sulfate (Sodium Sulfate) 100mM 0.39 0.41 0.80 0.54 1.41 1.95 0.75 2.49 3.24
A-6 Oh, unlike mumap 50mg / mL, PS801mg / mL, pH 5.2 no buffer/no salt 0.37 0.40 0.77 0.28 1.21 1.49 0.38 2.03 2.41
A-7 Sodium phosphate / sodium citrate (Humira Composition Buffer) * 0.38 0.41 0.79 0.43 1.27 1.69 0.66 2.20 2.86
A-8 NaCl 100mM 0.38 0.42 0.80 0.43 1.38 1.81 0.63 2.50 3.13
A-9 Ammonium sulfate (Ammonium sulfate) 100mM 0.39 0.41 0.80 0.38 1.45 1.83 0.53 2.58 3.10
A-10 Sodium sulfate (Sodium Sulfate) 100mM 0.39 0.44 0.83 0.40 1.45 1.85 0.55 2.51 3.05

[174]
*Humira 조성 Buffer: Sodium phosphate monobasic dihydrate 0.86 mg/mL, Sodium phosphate dibasic dihydrate 1.53 mg/mL, Sodium citrate 0.3 mg/mL, Citric acid monohydrate 1.3 mg/mL
[175]
[176]
Table 13 Formulation and zerotime, 40 of each sample of the sample o after storage C 2 ju, 40 o C 1 gaewol exhibited a SE-HPLC analysis after storage. Oh otherwise mumap 100mg / mL, and so that the PS80 1mg / mL was prepared the samples of A-1 where a buffer or salt to prepare the A-2 ~ A-5 to include a. In addition, by controlling the A-1 ~ A-5 O mumap different concentration of the composition to 50mg / mL to prepare a A-6 ~ A-10. In the SE-HPLC results for each sample, as compared to A-2, including the A-1 and a buffer that does not contain the buffer solution, (or Ah contrast when comparing the A-6 and A-7, unlike the mumap concentration ) the formulation does not contain a buffer it was smaller the HMW and LMW growth rate, and thus it can be seen that a more stable formulation.
[177]
In addition, A-1 and A-6 and, NaCl, ammonium sulfate when compared to formulations containing the salts of (Ammonium Sulfate), sodium sulphate (Sodium Sulfate) when the formulation does not contain a salt, as compared with the formulation containing the salt HMW and a small increment of LMW. Thus, unlike Oh Oh that otherwise do not contain the salt in the case of mumap formulation can be seen better return on the stability of mumap.
[178]
In addition, the pH of all samples was held constant at around the storage, archiving and storage 5.2 2 weeks after one month later. In contrast ah than 50mg / mL, depending was determined that mumap have a sufficient self-buffering effect, and therefore no need for the use of a buffer, and it was confirmed that it is possible to configure a formulation having a higher reliability by not using a buffer.
[179]
[180]
< Example 10>
[181]
Oh otherwise mumap confirmation buffer and salt effects on the formulation Test 2
[182]
[183]
Oh otherwise otherwise Ah to identify the buffer and salt effects on the stability of mumap mumap (100mg / mL, or 50mg / mL), a stabilizer (sucrose, 55mg / mL or glycine 160mM), arginine hydrochloride (ArgHCl) 50mM, methionine 5mM and poly sorbic was prepared a sample of the bait formulation 80 consisting of a 1mg / mL, a sample of the formulation by the addition of a buffer or salt was prepared to the formulation. For comparison Oh otherwise mumap 100 mg / mL, or otherwise O mumap 50mg / mL was prepared for a sample of HUMIRA composition, each dosage form 55 is filled by a glass syringe 0.4mL o 1 ju storage stability to a SE-HPLC in C They were compared to measure the pH. In the table below for each composition and the 55 o exhibited a C 1 ju storage before / after monomer (monomer) content.
[184]
[Table 14]
[185]
[186]
*Humira 조성: Sodium phosphate monobasic dihydrate 0.86 mg/mL, Sodium phosphate dibasic dihydrate 1.53 mg/mL, Sodium citrate 0.3 mg/mL, Citric acid monohydrate 1.3 mg/mL, Mannitol 12 mg/mL, Sodium chloride 6.16 mg/mL, PS80 1 mg/mL
[187]
**Humira 조성 Buffer: Sodium phosphate monobasic dihydrate 0.86 mg/mL, Sodium phosphate dibasic dihydrate 1.53 mg/mL, Sodium citrate 0.3 mg/mL, Citric acid monohydrate 1.3 mg/mL
[188]
[189]
First, pH of all samples was 55 o C 1 ju storage before / after was kept constant at 5.2. Accordingly, it was confirmed that the composition in the table, and otherwise similar to the composition Oh mumap least 50mg / mL have a buffering effect in itself sufficient.
[190]
As the results in Table 14, stored total monomer content of the samples was similar to 98.8 ~ 99.0%. 55 o C 1 ju after storage were the monomer content differently for each composition. Oh otherwise mumap 100mg / mL, sucrose, 55mg / mL, ArgHCl 50mM, methionine 5mM and polysorbate 80 1mg / mL samples (A-11) of the following composition is ammonium monomer content is yieoteuna 94.8% NaCl, after storage of sulfuric acid, MgCl 2 and CaCl 2Salts, or, if using a sodium acetate buffer (A-12 ~ A-16) The monomer content was lower than that of the sample did not receive the content after storage of the monomer used and the salt buffer to 94.1 ~ 94.7% composition. When using the Gly 160mM place of sucrose stabilizer (A-17) it inde a 95.2% content of the store after the monomer salt, and (A-18 ~ A-23) kept after the monomer content of the salt to 94.0 ~ 94.9% when using the buffer than and the monomer content was lower than the compositions did not use a buffer. Oh otherwise, even if joseonghan lower the content of mumap to 50mg / mL when using sucrose as a stabilizer without the use of salts and buffers, if after the monomer content with 95.2% (A-25), Glycine (Gly) as a stabilizer Storage Storage after the monomer content of 95.6% (a-32) yieoteuna Add salt / storage after 93.5 ~ 95.0% monomer when the content of the sucrose containing formulations (a-26 ~ a-31) the case of using a buffer in each of the formulations, Gly contain formulation when (a-33 - a-38) was lower than the 93.6-monomer content of not using the additional salt / formulations of the monomer content of the salt / buffer with a buffering agent to a 95.4% formulation. Therefore, ah, unlike the additional buffer with a buffering effect of mumap, arginine, stabilizers Oh that agent and that does not use a dosage form additional salts and buffering agents in containing a surface active agent contrast was confirmed by increasing the stability of mumap, Oh otherwise it mumap itself it was confirmed that it is possible to configure that the composition without increasing the stability.
[191]
However HUMIRA the same Oh otherwise mumap content composition samples (A-24, A-39 ) of the formulation containing the salt and a buffering agent when compared to the 55 o C 1 ju stored after the monomer content of a HUMIRA composition sample 55 o C 1 ju storage It was over after the monomer content.
[192]
Accordingly, arginine, surfactants, Oh otherwise includes one or desirable in terms of stability is not used for more salts and buffers to mumap formulations, arginine, surface active agent, adding salt and buffer the formulation comprises a stabilizer or without containing stabilizer without a correlation was confirmed that HUMIRA stable than the commercial formulation.
[193]
[194]

[195]
Oh sure, unlike stabilizing effect of polyols in formulation experiments mumap
[196]
[197]
Oh contrast to the Ah otherwise mumap 112mg / mL solution and a Oh otherwise mumap 112mg / mL, and the formulation containing 42mg / mL of each polyol in order to compare the stabilizing effect of polyols is used to promote in stable solution of mumap prepared as follows: -70 o C and 5 o from the C repeats the cycle 5, and the same cold weather 10cycle analyzed by SE-HPLC and analyzed the content of the HMW, LMW and monomer.
[198]
[Table 15]
Formulation Zerotime FT 5C FT 10C
HMW (%) LMW (%) Total(%) HMW(%) LMW (%) Total(%) HMW(%) LMW (%) Total(%)
No polyol 0.40 0.42 0.82 1.63 0.49 2.13 2.08 0.37 2.45
Mannitol 42mg / mL 0.39 0.41 0.80 0.51 0.48 0.99 0.65 0.36 1.01
Sucrose, 42mg / mL 0.38 0.41 0.79 0.35 0.47 0.82 0.34 0.34 0.68
Trehalose 42mg / mL 0.38 0.41 0.79 0.35 0.47 0.82 0.35 0.35 0.70

[199]
sampling point of the formulation and the stability test of samples in TABLE 15 are shown by SE-HPLC results. When repeating the cold weather move the respective samples in the above results, shown mannitol, sucrose, or as compared to formulations that are not the case of the formulation added to the trehalose was added to the polyol tends to HMW and LMW increase reduced, the stabilizing effect of the polyol was confirmed that the. Comparing the type stabilizing effect of each polyol, if you have the sucrose or trehalose similar to the content of the sample and the HMW and LMW before cold weather copper, cold weather copper 10 cycle after it was confirmed stabilized show a similar purity as before cold weather the same effect is great. On the other hand, if the sample including mannitol when repeating the same cold weather tended to the HMW content increases, it was confirmed that the purity becomes lower during the cold weather during the same period. As a result, the stabilization effect of sucrose and trehalose was confirmed the superiority compared to the mannitol accordingly.
[200]
[201]

[202]
Arginine, methionine, O otherwise mumap formulations such cold weather test for a stabilizing effect and make comparison of glycine and sucrose
[203]
[204]
Arginine, methionine, glycine and sucrose is prepared of the sample immediately after preparation and then 5mL polycarbonate containing 1mL by the bottle and -70 as follows: In order to confirm the effects of cold weather simultaneous stabilization of the stock solution o C, 5 o the cold weather in the same C after repeating 5 times were analyzed by SE-HPLC. The composition and the same cold weather SE-HPLC results before / after the respective samples are as follows.
[205]
[Table 16]
Furtherance Add stabilizer All such cold weather After five such cold weather
ArgHCl(mM) Gly(mM) Met(mM) Sucrose (mg / mL)
HMW LMW Total HMW LMW Total
Oh otherwise mumap 130mg / mL, polysorbate 80 1mg / mL - - -　 -　 0.32 0.40 0.72 1.76 0.28 2.04
50 -　 -　 -　 0.36 0.39 0.74 0.67 0.31 0.98
50 -　 5 -　 0.34 0.37 0.71 0.67 0.33 1.00
50 -　 25 -　 0.33 0.34 0.67 0.52 0.33 0.85
50 160 5 -　 0.34 0.36 0.70 0.41 0.31 0.73
50 140 25 -　 0.33 0.34 0.66 0.43 0.33 0.75
50 0 5 55 0.34 0.35 0.69 0.41 0.29 0.70

[206]
Oh otherwise mumap 130mg / mL, poly sorbitan samples were prepared so that the bait 80 1mg / mL, to thereby prepare a sample to contain an additional stabilizer to it. Impurity content of each sample of the cold weather coins were similar both HMW and LMW. After such cold weather, LMW for all samples was similar showed the amount of HMW different depending on the type and amount of stabilizer added. If you do not include additional stabilizers such cold weather hoehu 5 is HMW was increased to 1.76%, the same cold weather after five HMW was greatly reduced to 0.67% when arginine hydrochloride containing 50mM. For the formulation containing an additional methionine to arginine hydrochloride 50mM, when the methionine 5mM hayeoteul include, but could not confirm the reduction of the additional HMW, using 25mM there was confirmed that the HMW is further reduced, so 0.52%. If you add the addition of glycine, or sucrose as the arginine hydrochloride, methionine cold weather copper was reduced to 5 times after the HMW further reduced 0.41 ~ 0.43%, it was confirmed that the glycine-containing formulations with sucrose containing formulations showing a similar stability. Thus, methionine, arginine, glycine, it was confirmed that the sucrose have all contributed to the stability of mumap otherwise H. Also was confirmed that by using a polyol or an amino acid as a stabilizer can be obtained, the stabilizing effect is similar when the right combination.
[207]

[208]
Glycine containing formulations, the formulation containing sucrose and Humira stability of the commercial formulation Comparative test
[209]
[210]
Glycine containing formulations, sucrose include the formulation and Humira otherwise ah order to compare the stability of the commercial formulation content of mumap to be 100mg / mL, or 50mg / mL, arginine hydrochloride (ArgHCl) 50mM, polysorbate 80 (PS80) 1mg / mL to prepare a sample so as to include a methionine and 5mM prepare a sample using a combination, or of sucrose, glycine, glycine and methionine stabilizing agent added to the composition. Also, unlike the O content of the sample was prepared mumap as HUMIRA commercial composition such that the 100mg / mL, or 50mg / mL. Each of the filled by 0.4mL in 1mL glass syringe 40 o analyzed the HMW and LMW content of C 4 ju storage before / after by SE-HPLC. The results of each composition and SE-HPLC is as follows.
[211]
[212]
[Table 17]
Furtherance Add stabilizer 40 O C 4 weeks ago archived 40 O C 4 weeks of storage
HMW LMW Total HMW LMW Total
Oh otherwise mumap 100mg / mL, ArgHCl 50mM, PS80 1mg / mL Met 5mM - 0.28 0.39 0.67 0.54 2.44 2.99
Gly 120mM 0.27 0.41 0.68 0.56 2.42 2.99
Gly 160mM 0.28 0.42 0.69 0.54 2.37 2.91
Gly 100mM, Met 20mM 0.26 0.40 0.67 0.54 2.40 2.94
Gly 120mM, Met 20mM 0.27 0.39 0.65 0.53 2.46 2.99
Gly 140mM, Met 20mM 0.28 0.43 0.70 0.49 2.32 2.81
Sucrose, 55mg / mL 0.27 0.40 0.67 0.48 2.51 2.99
Oh otherwise mumap 100mg / mL, Humira composition 0.40 0.40 0.80 0.98 3.05 4.03
Oh otherwise mumap 50mg / mL, ArgHCl 50mM, PS80 1mg / mL, Met 5mM - 0.27 0.38 0.64 0.35 2.36 2.71
Gly 120mM 0.26 0.38 0.64 0.35 2.29 2.64
Gly 160mM 0.28 0.43 0.71 0.32 2.26 2.58
Gly 100mM, Met 20mM 0.26 0.39 0.64 0.33 2.33 2.66
Gly 120mM, Met 20mM 0.25 0.36 0.61 0.34 2.38 2.71
Gly 140mM, Met 20mM 0.27 0.43 0.70 0.30 2.26 2.56
Sucrose, 55mg / mL 0.26 0.38 0.64 0.31 2.33 2.64
Oh otherwise mumap 50mg / mL, Humira composition 0.43 0.41 0.84 0.82 3.23 4.06

[213]
Arginine hydrochloride, polysorbate 80, methionine, O, unlike the sample of the composition containing the mumap are 40 o exhibited similar stability C 4 ju stored before. The samples of the composition are the high degree of commercial Humira HMW 0.1% than the other sample immediately after manufacture. After storage the sum of the HMW and LMW content is relatively low ateuna HUMIRA commercial composition sample at 2.81 ~ 2.99%, O 2.56 ~ 2.71% when mumap 50mg / mL contrast, when all the samples for contrast ah mumap 100mg / mL day, including arginine hydrochloride in other cases, Oh was higher than the sample content of HMW and LMW in the composition comprises arginine in 4.06% at 4.03%, O otherwise mumap 50mg / mL when mumap 100mg / mL day. Accordingly, it can be seen that the combination of additives described in the present embodiment, that is, the dosage form comprising the formulation, and additional arginine, polyol or amino acid stabilizer comprising arginine formulation superior in stability than if commercial Humira formulation.
[214]
[215]
< Example 14 >
[216]
Oh otherwise mumap , sucrose, glycine, leucine , methionine, sodium chloride, the concentration of arginine-specific ah otherwise mumap stability compared to the test formulation
[217]
[218]
O In order to compare the stability of the otherwise mumap formulation, Oh otherwise mumap, arginine hydrochloride (ArgHCl), sodium chloride (NaCl), polysorbate 80 (PS80), methionine (Met), sucrose (Sucrose), glycine (Gly), leucine a combination of (Leu) to prepare a sample with a variable composition. In addition, unlike O For the purpose of comparison the amount of mumap to prepare a sample with Humira commercial composition such that the 100mg / mL, or 50mg / mL. 40 is filled by 0.4mL of each formulation in 1mL glass syringe o to C 4 ju storage before / after the monomer content were analyzed by SE-HPLC. The results of each composition and SE-HPLC is as follows.
[219]
[Table 18]
　 Furtherance Monomer content (%)
Oh otherwise mumap (mg / mL) ArgHCl (mM) NaCl (mM) Met (mM) PS80 (mg/mL) Sucrose (mg/mL) Gly (mM) Leu (mM) 40 ° C 4 weeks ago archived 40 ° C 4 weeks of storage
A-40 100 50 　 5 1 55 　 　 99.29 97.15
A-41 100 50 　 5 1 45 　 　 99.33 97.16
A-42 100 50 　 25 1 45 　 　 99.30 97.14
A-43 100 50 　 5 1 45 20 　 99.30 97.18
A-44 100 50 　 5 1 45 　 20 99.30 97.12
A-45 100 50 　 5 1 35 　 　 99.30 97.06
A-46 100 50 　 25 1 35 　 　 99.30 97.14
A-47 100 50 　 25 1 35 40 　 99.33 97.09
A-48 100 50 　 25 1 35 20 20 99.29 97.23
A-49 100 50 　 5 1 25 　 　 99.31 97.11
A-50 100 50 　 25 1 25 　 　 99.32 97.13
A-51 100 50 　 25 1 25 60 　 99.30 97.17
A-52 100 50 　 25 1 25 40 20 99.31 97.17
A-53 100 50 　 5 1 　 　 　 99.31 97.06
A-54 100 50 　 25 1 　 　 　 99.32 97.13
A-55 100 50 　 25 1 　 140 　 99.30 97.04
A-56 100 50 50 5 1 25 　 　 99.31 96.96
A-57 100 50 50 15 1 25 　 　 99.28 96.93
A-58 100 50 50 25 1 25 　 　 99.27 96.89
A-59 100 25 60 5 1 25 　 　 99.30 96.88
A-60 100 25 60 35 1 25 　 　 99.29 97.00
A-61 100 25 60 5 1 35 　 　 99.27 96.85
A-62 100 Commercial formulations HUMIRA 99.29 95.64
A-63 50 50 　 5 1 55 　 　 99.30 97.23
A-64 50 50 　 5 1 45 　 　 99.30 97.21
A-65 50 50 　 25 1 45 　 　 99.31 97.28
A-66 50 50 　 5 1 45 20 　 99.30 97.30
A-67 50 50 　 5 1 45 　 20 99.30 97.32
A-68 50 50 　 5 1 35 　 　 99.30 97.23
A-69 50 50 　 25 1 35 　 　 99.31 97.23
A-70 50 50 　 25 1 35 40 　 99.30 97.27
A-71 50 50 　 25 1 35 20 20 99.30 97.34
A-72 50 50 　 5 1 25 　 　 99.31 97.15
A-73 50 50 　 25 1 25 　 　 99.30 97.24
A-74 50 50 　 25 1 25 60 　 99.30 97.32
A-75 50 50 　 25 1 25 40 20 99.31 97.26
A-76 50 50 　 5 1 　 　 　 99.29 97.25
A-77 50 50 　 25 1 　 　 　 99.30 97.33
A-78 50 50 　 25 1 　 140 　 99.29 97.33
A-79 50 50 　 25 1 　 120 20 99.31 97.38
A-80 50 50 50 5 1 25 　 　 99.29 97.06
A-81 50 50 50 15 1 25 　 　 99.30 96.90
A-82 50 50 50 25 1 25 　 　 99.27 97.02
A-83 50 25 60 5 1 25 　 　 99.30 96.81
A-84 50 25 60 35 1 25 　 　 99.30 97.00
A-85 50 25 60 5 1 35 　 　 99.29 96.92
A-86 50 Commercial formulations HUMIRA 99.29 95.47

[220]
*상용 휴미라 제형: Sodium phosphate monobasic dihydrate 0.86 mg/mL, Sodium phosphate dibasic dihydrate 1.53 mg/mL, Sodium citrate 0.3 mg/mL, Citric acid monohydrate 1.3 mg/mL, Mannitol 12 mg/mL, Sodium chloride 6.16 mg/mL, PS80 1 mg/mL
[221]
Monomer content of all samples was similar irrespective of the formulation prior to storage 99.27% ​​~ 99.33% at 40 ° C.
[222]
Oh otherwise 40 ° C 4 weeks of storage the monomer content of the content of mumap 100mg / mL in sample (A-40 ~ A-61) is, in the case of which does not contain sodium chloride formulations (A-40 ~ A-55) 97.04 % - it was 97.23%, formulations containing sodium chloride (a-56 - a-61, a commercial Humira except composition a-62) slightly lower than when the formulation does not contain sodium chloride in 96.85% - 97.00% of. However, sodium chloride, sucrose, methionine, glycine, 40 ° C 4 weeks after storage monomer content differences in the content of leucine was relatively insignificant, the same monomer content of the composition (A-40 ~ A-61), including arginine O Unlike was confirmed that at least 1% higher than the monomer content of 95.64% of the commercial Humira composition (a-62) of mumap concentration.
[223]
Oh otherwise, even if the mumap content of 50mg / mL contrast O 100mg / mL are shown results similar to the result of the analysis of mumap composition. Oh otherwise 40 ° C 4 weeks of storage the monomer content of the content of mumap 50mg / mL in sample (A-63 ~ A-85) is, in the case of which does not contain sodium chloride formulations (A-63 ~ A-79) 97.15 % - it was 97.38%, formulations containing sodium chloride (a-80 - a-85, a commercial Humira except composition a-86) slightly lower than when the formulation does not contain sodium chloride in 96.81% - 97.06% of. However, sodium chloride, sucrose, methionine, glycine, 40 ° C 4 main monomer content differences after storage according to the content of leucine was relatively insignificant, the same monomer content of the composition (A-63 ~ A-85), including arginine O Unlike was confirmed that at least 1% higher than the monomer content of 95.47% of the commercial Humira composition (a-86) of mumap concentration.
[224]
As a result, it was confirmed that the combination and the like of the additive formulation described in this embodiment, that is, formulations containing arginine formulation superior in stability than if commercial Humira formulation.
[225]
[226]
From the above description, those skilled in the art will appreciate that may be embodied in other specific forms of the present invention without changing the technical spirit or essential characteristics. The embodiments described above In this regard, the examples should be understood as illustrative and not be limiting in all aspects. The scope of the invention should be construed as the meaning and scope, and all such modifications as derived from the equivalent concepts of the claims to be described later, rather than the description above within the scope of the invention.

Claims

[Claim 1]Wherein the liquid formulation of the -TNF alpha antibody comprising an anti -TNF alpha antibody, stabilizers, surfactants, and arginine.
[Claim 2]
The method of claim 1 wherein the liquid preparation of the stabilizing agent is a polyol, other amino acid other than arginine, or a combination thereof, wherein -TNF alpha antibody.
[Claim 3]
The method of claim 2 wherein the liquid formulation of, anti -TNF alpha antibodies that said polyol is selected from the group consisting of sucrose, trehalose, PEG, and combinations thereof.
[Claim 4]
The method of claim 2, wherein the amino acid is in, wherein the liquid formulation of the -TNF alpha antibody is selected from glycine, leucine, isoleucine, phenylalanine, proline and combinations thereof.
[Claim 5]
The method of claim 1 wherein the stabilizing agent is (i) sucrose or trehalose, (ii) a number average molecular weight of from 200 to 600 of PEG, or a number average molecular weight of from 1000 to 8000 PEG, (iii) glycine or leucine, and ( iv) a liquid formulation of the above (i) to (iii) one of the two or more combinations selected from the group consisting of, wherein -TNF alpha antibody.
[Claim 6]
According to claim 2, wherein -TNF liquid preparations of alpha antibody in the formulation polyol is present at a concentration of 0.1 to 100 mg / mL.
[Claim 7]
The method of claim 2 wherein the liquid formulation of the amino acid other than arginine in the formulation is present at a concentration of 1 to 300 mM, wherein -TNF alpha antibody.
[Claim 8]
The method of claim 1 wherein the liquid formulation of the surfactant is a nonionic surfactant, an anti -TNF alpha antibody.
[Claim 9]
The method of claim 1 wherein the surfactant is a liquid formulation of a polysorbate or a poloxamer, wherein -TNF alpha antibody.
[Claim 10]
The method of claim 9 wherein the surfactant is a liquid formulation of the polysorbate 80, polysorbate 20, poloxamer 188, or phosphorus, wherein -TNF alpha antibody.
[Claim 11]
2. The method of claim 1, wherein -TNF liquid preparations of alpha antibody in the formulation surfactants are present at a concentration of 0.1 to 5 mg / mL.
[Claim 12]
The method of claim 1, wherein the arginine is in the liquid formulations of, anti -TNF alpha antibody salt form.
[Claim 13]
The method of claim 12, wherein the arginine is arginine hydrochloride (arginine hydrochloride) in the form of liquid preparations, the anti -TNF alpha antibody.
[Claim 14]
2. The method of claim 1, wherein -TNF liquid preparations of alpha antibody in the formulation arginine is present at a concentration of 0.1 to 200mM.
[Claim 15]
The method of claim 1 wherein the liquid formulation of the anti-alpha antibody is -TNF Oh otherwise mumap (adalimumab), anti -TNF alpha antibody.
[Claim 16]
The method of claim 1 wherein the liquid formulation of the anti -TNF alpha antibody in the formulation is present at a concentration of 1 to 250 mg / mL, wherein -TNF alpha antibody.
[Claim 17]
2. The method of claim 1, wherein -TNF liquid preparations of alpha antibody anti -TNF alpha antibody in the formulation is present at from 50 to 200 mg / mL concentration.
[Claim 18]
The method of claim 1 wherein the liquid formulation, the liquid formulation further comprises an antioxidant.
[Claim 19]
19. The method of claim 18 wherein the liquid formulation of the antioxidant is methionine in, wherein -TNF alpha antibody.
[Claim 20]
20. The method of claim 19,, wherein -TNF liquid preparations of alpha antibody in the preparation of methionine is present at a concentration of 1 to 50 mM.
[Claim 21]
Claim 1 to a liquid formulation of, wherein -TNF alpha antibody of claim 20 according to any one of claims, wherein the liquid formulation does not comprise additional salts and buffer.
[Claim 22]
Liquid formulations of any one of claims 1 to A method according to any one of claim 20 wherein, the pH of the liquid formulations to 4 to 6, wherein -TNF alpha antibody.
[Claim 23]
The method of claim 1, wherein the liquid formulation is -TNF alpha antibody of 1 to 250mg / mL concentration; 0.1 to polyol of 100mg / mL concentration; 0.1, wherein the liquid formulation of the -TNF alpha antibody comprising the arginine of the surface active agent, and 0.1 to 200mM concentration of 5mg / mL concentration.
[Claim 24]
The method of claim 1, wherein the liquid formulation is -TNF alpha antibody of 1 to 250mg / mL concentration; 1 to another amino acid other than arginine concentration of 300mM; 0.1, wherein the liquid formulation of the -TNF alpha antibody comprising the arginine of the surface active agent, and 0.1 to 200mM concentration of 5mg / mL concentration.
[Claim 25]
Wherein -TNF alpha antibody, a stabilizer, a surfactant, and a method for producing a liquid formulation of claim 1 comprising the step of mixing of the arginine.