CN116059342A

CN116059342A - Liquid preparation containing recombinant fully human anti-IFNAR 1 monoclonal antibody and preparation method thereof

Info

Publication number: CN116059342A
Application number: CN202210986979.9A
Authority: CN
Inventors: 邹志通; 张鑫; 常志远; 边琳; 王威
Original assignee: Genrix Shanghai Biopharmaceutical Co ltd; Chongqing Zhixiang Jintai Biopharmaceutical Co ltd
Current assignee: Genrix Shanghai Biopharmaceutical Co ltd; Chongqing Zhixiang Jintai Biopharmaceutical Co ltd
Priority date: 2022-08-17
Filing date: 2022-08-17
Publication date: 2023-05-05

Abstract

The present application provides pharmaceutical formulations, such as liquid formulations, comprising a recombinant fully human anti-IFNAR 1 monoclonal antibody and an osmolality regulator. The recombinant fully human anti-IFNAR 1 monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 sequences. The pharmaceutical formulations are useful for the prevention or treatment of diseases mediated by human IFNAR1, for example autoimmune diseases, and may be administered by injection.

Description

Liquid preparation containing recombinant fully human anti-IFNAR 1 monoclonal antibody and preparation method thereof

Technical Field

The present application relates generally to the field of pharmaceutical formulations comprising antibodies. In particular, the present application relates to stable pharmaceutical formulations comprising monoclonal antibodies against human type I interferon receptor subunit 1 (IFNAR 1) and methods of making the same.

Background

Interferons (IFNs) include type I interferons, type II interferons, and type III interferons. Wherein human type I interferon comprises IFN-alpha, IFN-beta, IFN-epsilon, IFN-kappa, and IFN-omega. Generally, when viruses invade the body, fibroblasts and monocytes in the human body secrete various type I interferons to prevent and interfere with the DNA and RNA replication of the viruses. In addition, type I interferons have anti-tumor and immunomodulatory functions (Capobiananchi M.R., et al, 2015,Cytokine Growth Factor Rev, 26:103;Zitvogel L, et al, 2015,Nat Rev Immunol, 15:405).

All human type I interferons share a set of cell surface receptor complexes, i.e., ifnα/β receptor complexes, which include two transmembrane protein subunits, IFNAR1 and IFNAR 2. IFNAR1 and IFNAR2 are both essential for the realization of type I interferon function and affect both the high affinity (KD of about 10-11M) and specificity (Bekisz J, et al 2004,Growth Factors,22:243) of the receptor complex for different type I interferons.

In recent years, more studies suggest that type I interferons have strong immunomodulatory effects in adaptive immunity, including promotion of antibody secretion and support of functional activity and survival of T memory cells. In particular, IFN- α has been shown to promote maturation or activation of Dendritic Cells (DCs) (Santini S.M., et al, 2000, J. EXP. Med., 191:1777). Furthermore, various autoimmune diseases have been found to exhibit type I interferon overexpression. Among them, insulin Dependent Diabetes Mellitus (IDDM) and Systemic Lupus Erythematosus (SLE) are associated with increased expression of IFN- α, while IFN- β may be associated with Rheumatoid Arthritis (RA). Moreover, it has been reported that clinical type I IFN administration results in exacerbations of some autoimmune diseases (including psoriasis, multiple sclerosis, etc.), and may induce SLE-like symptoms in patients without a history of autoimmune disease. Furthermore, studies have shown that TMPD is effective in inducing systemic lupus erythematosus symptoms in normal mice, but not in mice deficient in IFNAR1 gene (naconones d.c., et al, 2007,Arthritis Rheum,56:3770). In BXSB model mice, anti-IFNAR 1 antibodies showed significant therapeutic effects in early disease (Baccala R., et al 2012, J.Immunol, 189:5876). Thus, inhibition of type I interferon receptors (IFNAR) may be beneficial in patients with certain autoimmune diseases, and there is a clinical need for drugs that are effective in inhibiting type I interferon receptors for the treatment of various autoimmune diseases, including systemic lupus erythematosus, psoriasis, multiple sclerosis or rheumatoid arthritis.

As with any protein, the biological activity of an antibody depends on the conformational integrity of at least the core amino acid sequence remaining intact while protecting the various functional groups of the protein from degradation. Both chemical and physical instability can lead to antibody degradation. Because antibodies are larger and more complex than traditional organic and inorganic drugs, the formulation of such antibodies presents particular problems. Antibody stability can be affected by a variety of factors, such as pH, temperature, repeated freeze/thaw cycles, and shear forces, among others. Active antibodies may be lost as a result of physical instabilities including denaturation, aggregation (soluble and insoluble aggregate formation), precipitation and adsorption, and chemical instabilities including, for example, racemization, hydrolysis and deamidation, and the like. Any of these instabilities can potentially result in the formation of antibody byproducts or derivatives having reduced biological activity, increased toxicity, and/or increased immunogenicity.

While the prior art indicates examples of many excipients that may be suitable for use in antibody formulations for the production of a particular antibody, it cannot be predicted which excipients should be added and the amount that should be added to overcome the particular instability problems that a particular antibody may have. Furthermore, it is difficult to find optimal conditions for maintaining the chemical and biological stability of a particular antibody within a particular formulation. In view of all the factors that can vary, finding suitable excipients and optimal conditions for formulating antibodies challenges.

Thus, there remains a need in the art for stable pharmaceutical formulations comprising antibodies against human type I interferon receptor subunit 1 (IFNAR 1).

Summary of The Invention

In one aspect, the present application provides a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of a recombinant fully human anti-IFNAR 1 monoclonal antibody (hereinafter simply referred to as "monoclonal antibody"), an osmolality adjusting agent selected from the group consisting of pharmaceutically acceptable salts of trehalose and/or arginine, optionally a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0, wherein the monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 sequences, and specifically binds human type I interferon receptor subunit 1 (IFNAR 1).

In one aspect, the present application provides a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of a recombinant fully human anti-IFNAR 1 monoclonal antibody (hereinafter simply referred to as "monoclonal antibody"), a buffer salt, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, optionally a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0, wherein the monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 sequences, and specifically binds human type I interferon receptor subunit 1 (IFNAR 1).

In one aspect, the present application provides a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of a recombinant fully human anti-IFNAR 1 monoclonal antibody (hereinafter simply referred to as "monoclonal antibody"), a buffer salt, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0, wherein the monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2, and LCDR3 sequences, and specifically binds human type I interferon receptor subunit 1 (IFNAR 1).

In some embodiments, the present application provides a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of the monoclonal antibody, from about 10mM to about 50mM of a buffer salt, an osmotic pressure modulator selected from the group consisting of trehalose and/or pharmaceutically acceptable salts of arginine, optionally from about 0.05mg/ml to about 1.0mg/ml of a nonionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0, a concentration of trehalose of from 0 to about 150mg/ml, and a concentration of pharmaceutically acceptable salts of arginine of from 0 to about 50mg/ml.

In another aspect, the present application provides the use of a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of said monoclonal antibody, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, optionally a non-ionic surfactant, and the balance water, in the manufacture of a medicament for the prevention or treatment of a disease mediated by human IFNAR1, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0.

In another aspect, the present application provides a method for preventing or treating a disease mediated by human IFNAR1 in a subject, comprising administering to the subject in need thereof a pharmaceutical composition, wherein the pharmaceutical composition comprises from about 5mg/ml to about 200mg/ml of the monoclonal antibody, an osmolality regulator selected from a pharmaceutically acceptable salt of trehalose and/or arginine, optionally a non-ionic surfactant, and balance water, wherein the pH of the pharmaceutical composition is from about 4.8 to about 6.0.

In another aspect, the present application provides a pharmaceutical composition for preventing or treating a human IFNAR1 mediated disease in a subject comprising from about 5mg/ml to about 200mg/ml of the monoclonal antibody, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, optionally a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0.

In one embodiment of the present application, the pharmaceutical composition further comprises a nonionic surfactant, such as a polysorbate surfactant.

In one embodiment of the present application, the osmolality adjusting agent is a mixture of pharmaceutically acceptable salts of trehalose and arginine.

Drawings

FIGS. 1A-1G illustrate the effect of osmolality adjusting agents on the stability of pharmaceutical formulations comprising GR1603 protein.

Figures 2A-2G illustrate the effect of pH on the stability of a pharmaceutical formulation comprising GR1603 protein in an acetate/trehalose containing formulation.

Figures 3A-3E illustrate the effect of pH on stability of a pharmaceutical formulation comprising GR1603 protein in a formulation containing acetate/arginine hydrochloride.

Figures 4A-4G illustrate the effect of buffer salt concentration on the stability of a pharmaceutical formulation comprising GR1603 protein.

FIGS. 5A-5G illustrate the effect of the presence of polysorbate 80 on the stability of pharmaceutical formulations comprising the GR1603 protein.

FIGS. 6A-6G illustrate the effect of the type of buffer salt on the stability of a pharmaceutical formulation comprising the GR1603 protein.

Figures 7A-7E illustrate the effect of different ratios of arginine hydrochloride to trehalose mixtures on the stability of pharmaceutical formulations comprising GR1603 protein.

Figures 8A-8E illustrate the effect of different amino acid-type osmolytes on the stability of a pharmaceutical formulation comprising GR1603 protein.

Figures 9A-9C illustrate the effect of polysorbate 80 concentration on stability of pharmaceutical formulations comprising GR1603 protein in a shaking experiment.

Fig. 10 illustrates the effect of polysorbate 80 concentration on stability of pharmaceutical formulations comprising GR1603 protein in freeze-thawing experiments.

Figure 11 illustrates the stability of a pharmaceutical formulation comprising a high concentration of GR1603 protein.

Detailed Description

Recombinant fully human anti-IFNAR 1 monoclonal antibody

The present application provides recombinant fully human anti-IFNAR 1 monoclonal antibodies that specifically bind to human IFNAR1 and comprise a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 sequences, characterized in that: the HCDR1 sequence is NYWVA (SEQ ID NO: 29), the HCDR2 sequence is IIYPGDSDTRYSPSFQG (SEQ ID NO: 30), the HCDR3 sequence is HDVTGYDY (SEQ ID NO: 31), the LCDR1 sequence is RASQNVGNYLN (SEQ ID NO: 41), the LCDR2 sequence is RASNLAS (SEQ ID NO: 42), and the LCDR3 sequence is QQMEHAPPT (SEQ ID NO: 43); alternatively, the HCDR1 sequence is NYWVA (SEQ ID NO: 29), the HCDR2 sequence is IIYPGDSDTRYSPSFQG (SEQ ID NO: 30), the HCDR3 sequence is HDVTGYDY (SEQ ID NO: 31), the LCDR1 sequence is RASQSVIGYYLA (SEQ ID NO: 44), the LCDR2 sequence is SVSTLAS (SEQ ID NO: 45), and the LCDR3 sequence is QQYYRFPIT (SEQ ID NO: 46); alternatively, the HCDR1 sequence is NYWMA (SEQ ID NO: 32), the HCDR2 sequence is IIYPSDSDTRYSPSFQG (SEQ ID NO: 33), the HCDR3 sequence is HDVEGYDY (SEQ ID NO: 34), the LCDR1 sequence is RASQNVGNYLN (SEQ ID NO: 41), the LCDR2 sequence is RASNLAS (SEQ ID NO: 42), and the LCDR3 sequence is QQMEHAPPT (SEQ ID NO: 43); alternatively, the HCDR1 sequence is NYWMA (SEQ ID NO: 32), the HCDR2 sequence is IIYPSDSDTRYSPSFQG (SEQ ID NO: 33), the HCDR3 sequence is HDVEGYDY (SEQ ID NO: 34), the LCDR1 sequence is RASQSVIGYYLA (SEQ ID NO: 44), the LCDR2 sequence is SVSTLAS (SEQ ID NO: 45), and the LCDR3 sequence is QQYYRFPIT (SEQ ID NO: 46); alternatively, the HCDR1 sequence is NYWVA (SEQ ID NO: 29), the HCDR2 sequence is IIYPSDSDTRYSPSFQG (SEQ ID NO: 33), the HCDR3 sequence is HDVHGYDY (SEQ ID NO: 35), the LCDR1 sequence is RASQNVSNYLN (SEQ ID NO: 47), the LCDR2 sequence is RASNLQS (SEQ ID NO: 48), and the LCDR3 sequence is QQMMDAPPT (SEQ ID NO: 49); alternatively, the HCDR1 sequence is NYWIG (SEQ ID NO: 36), the HCDR2 sequence is RIYPSDSNTSYSPSFQG (SEQ ID NO: 37), the HCDR3 sequence is DASSKTYDS (SEQ ID NO: 38), the LCDR1 sequence is SGSSSNIGTNAVN (SEQ ID NO: 50), the LCDR2 sequence is SKNQRPP (SEQ ID NO: 51), and the LCDR3 sequence is AAWDDSQNGYVV (SEQ ID NO: 52); alternatively, the HCDR1 sequence is NYWIG (SEQ ID NO: 36), the HCDR2 sequence is RIYPGDSYTRYSPSFQG (SEQ ID NO: 39), the HCDR3 sequence is DGAPAKGDFDY (SEQ ID NO: 40), the LCDR1 sequence is RASEGIGNHLN (SEQ ID NO: 53), the LCDR2 sequence is TASNLQS (SEQ ID NO: 54), and the LCDR3 sequence is QQTYITPLT (SEQ ID NO: 55); wherein the HCDR and LCDR sequences are defined according to Kabat.

The structure, amino acid sequence, nucleotide sequence, preparation method and biological activity of the recombinant fully human anti-IFNAR 1 monoclonal antibody of the present application have been described in chinese patent application No. 201610634601.7, the entire contents of which are hereby incorporated by reference in their entirety.

In some embodiments, the heavy chain variable region sequence of an antibody that specifically binds human IFNAR1 is SEQ ID NO. 19 and the light chain variable region sequence is SEQ ID NO. 21.

In some embodiments, the heavy chain variable region sequence of the antibody that specifically binds human IFNAR1 is SEQ ID NO. 19 and the light chain variable region sequence is SEQ ID NO. 22.

In some embodiments, the heavy chain variable region sequence of the antibody that specifically binds human IFNAR1 is SEQ ID NO. 20 and the light chain variable region sequence is SEQ ID NO. 21.

In some embodiments, the heavy chain variable region sequence of the antibody that specifically binds human IFNAR1 is SEQ ID NO. 20 and the light chain variable region sequence is SEQ ID NO. 22.

In some embodiments, the heavy chain variable region sequence of the antibody that specifically binds human IFNAR1 is SEQ ID NO. 23 and the light chain variable region sequence is SEQ ID NO. 26.

In some embodiments, the heavy chain variable region sequence of the antibody that specifically binds human IFNAR1 is SEQ ID NO. 24 and the light chain variable region sequence is SEQ ID NO. 27.

In some embodiments, the heavy chain variable region sequence of an antibody that specifically binds human IFNAR1 is SEQ ID NO. 25 and the light chain variable region sequence is SEQ ID NO. 28.

In some embodiments, the antibody that specifically binds to human IFNAR1 is a fully human full length antibody.

In some embodiments, the antibody that specifically binds human IFNAR1 further comprises a heavy chain constant region selected from the group consisting of subtype IgG1 (SEQ ID NO: 7), subtype IgG2 (SEQ ID NO: 8) or subtype IgG4 (SEQ ID NO: 9) and/or comprises a light chain constant region selected from the group consisting of subtype kappa (SEQ ID NO: 10) or subtype lambda (SEQ ID NO: 11).

In some embodiments, the antibody that specifically binds to human IFNAR1 antagonizes at least one biological activity associated with IFNAR1 or a portion thereof in vitro or in vivo.

In some embodiments, the antibody that specifically binds IFNAR1 is capable of specifically binding to the extracellular region of recombinant human IFNAR 1.

Pharmaceutical composition

In one aspect, the present application provides a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of a recombinant fully human anti-IFNAR 1 monoclonal antibody, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, optionally a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0, wherein the monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 sequences, and specifically binds human type I interferon receptor subunit 1 (IFNAR 1).

In one aspect, the present application provides a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of a recombinant fully human anti-IFNAR 1 monoclonal antibody, a buffer salt, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, optionally a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0, wherein the monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 sequences, and specifically binds human type I interferon receptor subunit 1 (IFNAR 1).

In one aspect, the present application provides a pharmaceutical composition comprising from about 5mg/ml to about 200mg/ml of a recombinant fully human anti-IFNAR 1 monoclonal antibody, a buffer salt, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 6.0, wherein the monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2, and LCDR3 sequences, and specifically binds human type I interferon receptor subunit 1 (IFNAR 1).

In some embodiments, the present application provides a pharmaceutical composition comprising from about 40mg/ml to about 170mg/ml of the monoclonal antibody, from about 10mM to about 50mM of a buffer salt, an osmotic pressure modulator selected from a pharmaceutically acceptable salt of arginine or a mixture of trehalose and a pharmaceutically acceptable salt of arginine, from about 0.1mg/ml to about 0.3mg/ml of a nonionic surfactant, and balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 5.6, a concentration of trehalose of from 0 to about 90mg/ml, and a concentration of a pharmaceutically acceptable salt of arginine of from about 10mg/ml to about 50mg/ml.

In some embodiments, the present application provides a pharmaceutical composition comprising from about 40mg/ml to about 150mg/ml of the monoclonal antibody, from about 10mM to about 50mM of a buffer salt, an osmolality adjuster selected from the group consisting of a pharmaceutically acceptable salt of arginine or a mixture of trehalose and a pharmaceutically acceptable salt of arginine, from about 0.1mg/ml to about 0.3mg/ml of a nonionic surfactant, and balance water, wherein the pharmaceutical composition has a pH of from about 4.8 to about 5.6, a concentration of trehalose of from 0 to about 90mg/ml, and a concentration of pharmaceutically acceptable salt of arginine of from about 10mg/ml to about 50mg/ml, wherein the amount of the pharmaceutically acceptable salt of arginine contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is from about 50 to about 95 parts by weight relative to 100 parts by weight of trehalose, and the amount of trehalose contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is from about 5 to about 50 parts by weight relative to 100 parts by weight of trehalose.

In some embodiments, the monoclonal antibody is present in the pharmaceutical composition in an amount of about 5mg/ml to about 200mg/ml, about 40mg/ml to about 170mg/ml, about 40mg/ml to about 150mg/ml, about 40mg/ml to about 120mg/ml, about 40mg/ml to about 100mg/ml, about 40mg/ml to about 60mg/ml, or about 45mg/ml to about 55mg/ml, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200mg/ml. Alternatively, one skilled in the art can select the appropriate amount of the monoclonal antibody according to the needs of the practice.

In some embodiments, the buffer salts may be conventional buffer salts used in the art to prepare liquid formulations, examples of which include, but are not limited to, phosphate buffer pairs, histidine/histidine inorganic salt buffer pairs, acetate buffer pairs, citrate buffer pairs, and the like. In some embodiments, the buffer salt is an acetic acid/acetate buffer pair, such as an acetic acid/sodium acetate buffer pair, an acetic acid/potassium acetate buffer pair, or an acetic acid/ammonium acetate buffer pair. In some embodiments, the acetic acid/acetate buffer pair is an acetic acid/acetate buffer system having a buffer range of ph3.8-ph 5.8.

In some embodiments, the concentration of the buffer salt in the pharmaceutical composition is from about 10mM to about 50mM, from about 10mM to about 45mM, from about 10mM to about 40mM, from about 10mM to about 35mM, from about 10mM to about 30mM, from about 15mM to about 25mM, from about 15mM to about 20mM, from about 20mM to about 25mM, or about 20mM.

In some embodiments, the buffer salt is an acetic acid/acetate buffer pair, e.g., an acetic acid/sodium acetate buffer pair, at a concentration of about 10mM to about 50mM, about 10mM to about 45mM, about 10mM to about 40mM, about 10mM to about 35mM, about 10mM to about 30mM, about 15mM to about 25mM, about 15mM to about 20mM, about 20mM to about 25mM, or about 20mM. Alternatively, one skilled in the art can select the appropriate concentration of acetic acid/acetate buffer pair as desired for practice.

In some embodiments, the pharmaceutical composition has a pH of not 5.8 when buffered with an inorganic salt of histidine/histidine (e.g., histidine hydrochloride), e.g., the pH may be 5.0, 5.2, 5.5, or 6.0.

In some embodiments, the osmolality adjusting agent is selected from the group consisting of trehalose alone, arginine hydrochloride (for example), or a mixture of trehalose and arginine hydrochloride (for example, arginine hydrochloride), preferably a mixture of trehalose and arginine. In some embodiments, the concentration of trehalose is from 0 to about 150mg/ml, preferably from 0 to about 90mg/ml, more preferably 17mg/ml, and the concentration of arginine hydrochloride is from 0 to about 50mg/ml, preferably from about 10mg/ml to about 30mg/ml, more preferably 24mg/ml.

In this application, pharmaceutically acceptable salts of trehalose and arginine (particularly arginine hydrochloride) are used as osmolality adjusting and stabilizing agents in the liquid formulations of the present application comprising recombinant fully human anti-IFNAR 1 monoclonal antibodies. The pharmaceutically acceptable salts of arginine, particularly arginine hydrochloride, can well inhibit aggregation and precipitation of the GR1603 protein while inhibiting the production of acidic variants. While arginine hydrochloride may cause proteolytic fragmentation of GR1603, the simultaneous addition of trehalose further promotes maintenance of GR1603 protein stability. Trehalose has a solvent exclusion effect in protein solutions, and can improve the stability of proteins in aqueous solutions.

In some embodiments, the amount of the pharmaceutically acceptable salt of arginine contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is about 50 to about 95 parts by weight, about 55 to about 90 parts by weight, about 60 to about 90 parts by weight, about 65 to about 85 parts by weight, about 70 to about 80 parts by weight, or about 75 parts by weight relative to 100 parts by weight of the pharmaceutically acceptable salt of arginine, and the amount of trehalose contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is about 5 to about 50 parts by weight, about 10 to about 45 parts by weight, about 10 to about 40 parts by weight, about 15 to about 35 parts by weight, about 20 to about 30 parts by weight, or about 25 parts by weight relative to 100 parts by weight of trehalose. For example, with 85mg/ml trehalose being 100% trehalose (i.e., 100 parts by weight trehalose) which is isotonic, the amount of trehalose contained in the mixture of trehalose and a pharmaceutically acceptable salt of arginine is 80% (68 mg/ml), 50% (42.5 mg/ml) or 20% (17 mg/ml), respectively. Arginine hydrochloride at an isotonic level of 30mg/ml is 100% arginine hydrochloride (i.e., 100 parts by weight of arginine hydrochloride), and the amount of the pharmaceutical salt of arginine (e.g., arginine hydrochloride) contained in the mixture of trehalose and the pharmaceutical salt of arginine is 80% (24 mg/ml), 50% (15 mg/ml), or 20% (6 mg/ml), respectively.

In some embodiments, in the pharmaceutical composition, the nonionic surfactant may be a polysorbate-type surfactant, such as polysorbate 20 or polysorbate 80 (i.e., polysorbate 80).

In some embodiments, the amount of nonionic surfactant (e.g., polysorbate-type surfactant, such as polysorbate 20 or polysorbate 80) is about 0.05mg/ml to about 1.0mg/ml, about 0.1mg/ml to about 0.3mg/ml, about 0.2mg/ml to about 0.5mg/ml, about 0.3mg/ml to about 1.0mg/ml, or about 0.5mg/ml to about 1.0mg/ml, such as about 0.2mg/ml, about 0.3mg/ml, about 0.34mg/ml, about 0.4mg/ml, about 0.5mg/ml, about 0.6mg/ml, about 0.7mg/ml, about 0.8mg/ml, about 0.9mg/ml, or about 1.0mg/ml.

In the present application, the monoclonal antibody may be denatured as a protein at an interface (air/liquid interface, liquid/glass interface, etc.) due to hydrophobic interaction, and thus protein aggregation and even protein precipitation may occur. The use of nonionic surfactants (e.g., polysorbate type surfactants such as polysorbate 20 or polysorbate 80) can prevent protein aggregation precipitation during shaking, agitation, or freeze thawing.

In some embodiments, the pharmaceutical composition has a pH of about 4.8 to about 6.0, about 5.0 to about 5.8, about 5.0 to about 5.6, about 5.0 to about 5.5, about 5.2 to about 5.6, or about 5.2 to about 5.5, e.g., the pH may be about 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0.

In some embodiments, the water in the pharmaceutical composition may be sterile, pyrogen-free water, such as water for injection.

In some embodiments, the present application provides a pharmaceutical composition comprising from about 40mg/ml to about 170mg/ml of the monoclonal antibody, from about 10mM to about 50mM of an acetic acid/acetate buffer pair, an osmolality adjuster selected from the group consisting of a pharmaceutically acceptable salt of arginine or a mixture of trehalose and a pharmaceutically acceptable salt of arginine, from about 0.1mg/ml to about 0.3mg/ml of a polysorbate surfactant, and balance water, wherein the pharmaceutical composition has a pH of from about 5.0 to about 5.5, a concentration of trehalose of from 0 to about 90mg/ml, and a concentration of a pharmaceutically acceptable salt of arginine of from about 10mg/ml to about 50mg/ml, wherein the amount of the pharmaceutically acceptable salt of arginine contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is from about 70 to about 90 parts by weight relative to 100 parts by weight of trehalose, and the amount of the trehalose contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is from about 10 to about 30 parts by weight relative to 100 parts by weight of trehalose.

In some embodiments, the present application provides a pharmaceutical composition comprising from about 40mg/ml to about 150mg/ml of the monoclonal antibody, from about 10mM to about 50mM of a acetic acid/sodium acetate buffer pair, an osmolality adjuster selected from the group consisting of a pharmaceutically acceptable salt of arginine or a mixture of trehalose and a pharmaceutically acceptable salt of arginine, from about 0.1mg/ml to about 0.3mg/ml of polysorbate 80, and balance water, wherein the pharmaceutical composition has a pH of from about 5.0 to about 5.5, a concentration of trehalose of from 0 to about 90mg/ml, and a concentration of a pharmaceutically acceptable salt of arginine of from about 10mg/ml to about 50mg/ml, wherein the amount of the pharmaceutically acceptable salt of arginine contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is from about 75 to about 95 parts by weight relative to 100 parts by weight of trehalose, and the amount of the trehalose contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is from about 15 to about 25 parts by weight relative to 100 parts by weight of trehalose.

In some embodiments, the present application provides a pharmaceutical composition comprising about 150mg/ml of the monoclonal antibody, about 20mM acetic acid/acetate (e.g., acetic acid/sodium acetate) buffer pair, an osmotic pressure regulator selected from the group consisting of a mixture of trehalose and a pharmaceutically acceptable salt of arginine (e.g., arginine hydrochloride), about 0.2mg/ml of a polysorbate surfactant, and the balance water, wherein the pharmaceutical composition has a pH of about 5.2, wherein the amount of the pharmaceutically acceptable salt of arginine contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is about 80 parts by weight relative to 100 parts by weight of trehalose, and the amount of the trehalose contained in the mixture of trehalose and the pharmaceutically acceptable salt of arginine is about 20 parts by weight relative to 100 parts by weight of trehalose.

The pharmaceutical compositions of the present application are liquid formulations, which may be in the form of solutions, emulsions or suspensions, preferably solutions. The pharmaceutical compositions of the present application may be administered to a subject in need thereof by a parenteral route of administration, such as an injection route of administration. Such injection routes of administration include, but are not limited to, subcutaneous injection (e.g., wound subcutaneous infiltration injection), intramuscular injection, intradermal injection, and the like.

The pharmaceutical composition comprising the monoclonal antibody has no substantial change in SEC-HPLC monomer peak area, CEX-HPLC acid peak area, CE-SDS (non-reducing method) and activity level after being placed at 37 ℃ for 4 weeks. The pharmaceutical composition containing the monoclonal antibody can be stored for a long time under the light-shielding condition of 5+/-3 ℃, can keep stable quality, and ensures the safety, effectiveness and uniformity of medicines.

In another aspect, the present application provides the use of a pharmaceutical composition as described herein in the manufacture of a medicament for the prevention or treatment of a disease mediated by human IFNAR 1.

In some embodiments, the disease is an autoimmune disease. In some embodiments, the autoimmune disease includes, but is not limited to, systemic lupus erythematosus, psoriasis, multiple sclerosis, rheumatoid arthritis.

It should be understood that the foregoing detailed description is only for the purpose of making the contents of the present application more clearly apparent to those skilled in the art, and is not intended to be limiting in any way. Various modifications and changes to the described embodiments will occur to those skilled in the art.

The following examples are for the purpose of illustration only and are not intended to limit the scope of the present application. In the following examples, GR1603 protein is used as an example of the monoclonal antibodies of the present application. The sequence of GR1603 protein is disclosed in Chinese patent application No. 201610634601.7, the HCDR1 of the antibody is NYWMA (SEQ ID NO: 32), HCDR2 is IIYPSDSDTRYSPSFQG (SEQ ID NO: 33), and HCDR3 is HDVEGYDY (SEQ ID NO: 34); LCDR1 is RASQSVIGYYLA (SEQ ID NO: 44), LCDR2 is SVSTLAS (SEQ ID NO: 45), and LCDR3 is QQYYRFPIT (SEQ ID NO: 46); the heavy chain constant region of the IgG1 subtype (SEQ ID NO: 7) and the light chain constant region of the kappa subtype (SEQ ID NO: 10).

Examples

Example 1: effect of osmolality regulator on stability of pharmaceutical formulations comprising GR1603 protein

And (3) taking a proper amount of GR1603 protein sample, replacing ultrafiltration liquid exchange into pure water, and measuring the protein concentration to be 73.7mg/ml to obtain the GR1603 protein aqueous solution. A 5× formulation buffer stock (containing 5-fold concentrations of buffer salt and osmolality adjusting agent, pH approximately 5.5) was prepared for each of the formulations shown in table 1. Then, 0.68ml of the aqueous GR1603 protein solution, 0.12ml of water, 0.2ml of the 5 Xpreparation buffer mother liquor and 2. Mu.L of the 10% aqueous polysorbate 80 solution were taken and mixed uniformly to prepare the osmotic pressure regulator of trehalose, sucrose, mannitol, sodium chloride and arginine hydrochloride, and the preparation sample containing no osmotic pressure regulator, respectively.

Table 1 prescription composition of liquid formulations

And (2) the following steps: indicating that the prescription contains the corresponding substances; -: indicating that the prescription does not contain the corresponding substances;

20mM acetate is a buffer system composed of acetic acid and sodium acetate and containing 20mM acetic acid/acetic acid ion

Samples of each formulation were stored at high temperature (37.+ -. 2 ℃) for stability testing to accelerate degradation of the GR1603 protein and sampled at day 0, week 1, week 2, week 3 and week 4, and the purity of the formulation samples was determined at each time point using the SEC-HPLC, CEX-HPLC, nrCE-SDS and rCE-SDS methods described below. The Tm values of each of the prescribed samples were determined using the DSF method to determine their conformational stability properties. The detection results are shown in the following table and fig. 1A to 1G.

The purity of SEC-HPLC (size exclusion chromatography) is determined by adopting a TSKgel G3000SWXL chromatographic column and a high performance liquid chromatograph, wherein the mobile phase is 50mM PB/300mM NaCl,pH 7.0 solution, the sample is diluted and injected, isocratic elution is carried out at a flow rate of 1.0ml/min, the elution is carried out for 15min, the detection is carried out at 280nm ultraviolet wavelength, and the peak area percentage of the monomer and the polymer is obtained by adopting a peak area normalization method.

CEX-HPLC (cation exchange chromatography) purity determination Using MabPac ^TM SCX-10 BioLC ^TM Analytical chromatographic column and high performance liquid chromatograph, mobile phase A is 20mM ACES, pH 6.8 solution, mobile phase B is 20mM MES+0.5M NaCl,pH 6.8 solution, dilute the sample, sample injection, gradient elution at a flow rate of 0.8ml/min, eluting for 40min, detecting at 280nm ultraviolet wavelength, and peak area normalization method is adopted to obtain peak area percentages of acid peak, main peak and alkaline peak.

The NrCE-SDS (sodium dodecyl sulfate capillary electrophoresis) and rCE-SDS methods were carried out by using a capillary electrophoresis apparatus, using an uncoated capillary column having an effective length of 20cm and SDS-MW Gel separation Gel, a column temperature of 25℃and a separation voltage of 15kV, and a detection wavelength of 214nm, and the purity of the sample was measured under non-reducing and reducing conditions, respectively.

DSF (differential scanning fluorescence) is measured by using a Real-Time PCR instrument, and the sample is diluted and mixed with fluorescent dye @

Orange Protein gel stain, life) and transferring the mixture into a PCR tube, setting a temperature raising program (25 ℃ to 95 ℃,1 ℃/min), collecting fluorescence in real time, recording data, deriving a fluorescence-temperature curve, and obtaining the highest peak of the curve after derivation, namely the Tm value.

TABLE 2 influence of osmolality regulator on Tm values of pharmaceutical formulations comprising GR1603 protein

Formulation number	Tm1 value (. Degree. C.)	Tm2 value (. Degree. C.)
			S1	54.6	66.1
S2	52.9	65.0
			S3	54.9	65.7
S4	54.0	65.5
			S5	50.8	66.8
S6	49.6	63.4

TABLE 3 influence of osmolality adjusting agent on the content of related substances in pharmaceutical formulations comprising GR1603 protein

As can be seen from tables 2-3 and fig. 1A-1G, trehalose, sucrose and mannitol can increase Tm1 values of GR1603 proteins; sodium chloride and arginine hydrochloride, in turn, lower the Tm1 value of the GR1603 protein. Under the high temperature condition, arginine hydrochloride can obviously inhibit the increase of GR1603 protein polymer and inhibit the protein precipitation phenomenon of GR1603 protein, but can accelerate the fragmentation speed of GR1603 protein; arginine hydrochloride inhibits the increase of the acidic peak to some extent, which can significantly reduce the rate of increase of SEC-HPLC multimers and CEX-HPLC acidic variants; arginine hydrochloride accelerates GR1603 hydrolysis; and the effect of each osmolyte regulator on the change in purity of the GR1603 protein reduced CE-SDS was not significantly different.

GR1603 is an IgG1 subtype antibody, and it has been reported in the literature that the hinge region of an IgG1 subtype antibody is relatively flexible, and is susceptible to cleavage under high temperature conditions, producing fab+fc fragments and Fab fragments, which significantly reduce the rate of chemical degradation at 2-8 ℃. Without the rate of protein aggregation being slowed down much by the reduced temperature. Thus, inhibiting aggregation of the GR1603 protein should be prioritized when balancing whether or not specific excipients need to be added to the formulation to increase the stability of the GR1603 protein. Thus, arginine hydrochloride needs to be added to GR1603 formulations as an osmotic pressure regulator.

Example 2 influence of formulation pH on stability of pharmaceutical formulations comprising GR1603 protein

In a 20mM acetate system, liquid formulations were prepared using the preparation method described in example 1 and according to the prescription composition shown in the following table, except that the pH of the formulation was adjusted to 4.8 to 5.8.

Table 4 prescription composition of liquid formulations

And (2) the following steps: indicating that the prescription contains the corresponding substances;

The purity of each formulation sample was tested using storage conditions, sampling time points and test items similar to those described in example 1. The detection results are shown in the following table and fig. 2A to 2G.

Table 5 effect of pH of formulation on Tm of pharmaceutical formulation comprising GR1603 protein

Formulation number	Tm1 value (. Degree. C.)	Tm2 value (. Degree. C.)
			S1	54.6	66.1
S7	54.7	66.9
			S8	54.9	66.7
S9	55.0	66.5
			S10	55.0	65.8

Table 6 effect of pH of formulations on content of related substances in pharmaceutical formulations comprising GR1603 protein

As can be seen from the above table and fig. 2A to 2G, tm1 value of GR1603 protein is less affected by formulation pH, but Tm2 value gradually decreases with increasing formulation pH. Under high temperature conditions, the rate of protein polymer increase increases with increasing formulation pH; the pH of the preparation has no effect on the increasing speed of the acid peak and has no effect on the purity change of the non-reducing CE-SDS of the GR1603 protein; when the pH of the preparation is less than or equal to 5.2, protein precipitation phenomenon can not occur; at a pH of 4.8, the alkaline peak increases faster; the preparation has no obvious influence on CEX purity change when the pH value is 5.0-5.8; the purity of the reduction CE-SDS drops faster at the pH of the preparation of 5.8; the preparation has no influence on the purity change of GR1603 protein reduced CE-SDS at pH 4.8-5.5.

In a 20mM acetate system, a liquid preparation was prepared by the preparation method described in example 1 and according to the prescription composition shown in the following table, except that the pH of the preparation was adjusted to 5.0 to 5.6 and arginine hydrochloride was used as an osmotic pressure regulator.

TABLE 7 prescription composition of liquid formulations

The purity of each formulation sample was tested using storage conditions, sampling time points and test items similar to those described in example 1. The results of the detection are shown in the following table and fig. 3A to 3E.

TABLE 8 influence of pH of formulations with arginine hydrochloride as osmotic pressure regulator on the content of related substances in pharmaceutical formulations comprising GR1603 protein

As can be seen from the above table and FIGS. 3A to 3E, there is no difference in the rate of increase of protein polymer and the change of CEX acidic basic peak at pH 5.0 to 5.6 in the system containing 20mM acetate/arginine hydrochloride at high temperature.

Example 3 Effect of buffer salt concentration on stability of pharmaceutical formulations comprising GR1603 protein

Liquid formulations were prepared using the preparation method described in example 1 and according to the prescription composition shown in the following table, except that the concentration of acetate in the formulation was adjusted to 10 to 30 mM.

Table 9 prescription composition of liquid formulations

/>

The purity of each formulation sample was tested using storage conditions, sampling time points and test items similar to those described in example 1. The detection results are shown in the following table and fig. 4A to 4G.

Table 10 effect of buffer salt concentration on Tm values of pharmaceutical formulations comprising GR1603 protein

Formulation number	Tm1 value (. Degree. C.)	Tm2 value (. Degree. C.)
			S1	54.6	66.1
S11	55.0	66.0
			S12	55.0	66.0

The effect of buffer salt concentration in the formulations of Table 11 on the content of related substances in pharmaceutical formulations comprising GR1603 protein

As can be seen from the above table and FIGS. 4A to 4G, the Tm value of the GR1603 protein is not affected when the acetate concentration is varied in the range of 10 to 30 mM. Under the high temperature condition, when the acetate concentration is changed within the range of 10-30 mM, the increasing speed of the protein polymer and the change trend of CEX-HPLC purity are not affected, and the change trend of non-reducing CE-SDS purity are not obviously different.

Example 4 influence of the presence of nonionic surfactant on the stability of pharmaceutical formulations comprising GR1603 protein

Liquid formulations were prepared using the preparation method described in example 1 and according to the prescription composition shown in the following table, except that polysorbate 80 was not added.

TABLE 12 prescription composition of liquid formulations

/>

The purity of each formulation sample was tested using storage conditions, sampling time points and test items similar to those described in example 1. The detection results are shown in the following table and fig. 5A to 5G.

TABLE 13 influence of the presence of nonionic surfactants on the Tm of pharmaceutical formulations comprising GR1603 protein

Formulation number	Tm1 value (. Degree. C.)	Tm2 value (. Degree. C.)
			S1	54.6	66.1
S13	54.5	65.8

TABLE 14 influence of the presence of nonionic surfactants on the content of related substances in pharmaceutical formulations comprising GR1603 protein

As can be seen from the above table and FIGS. 5A-5G, the presence or absence of polysorbate 80 does not affect the Tm of the GR1603 protein. The rate of increase of protein aggregates, the CEX purity profile, the non-reducing CE-SDS purity profile and the reducing CE-SDS purity profile are substantially unaffected by the presence or absence of polysorbate 80 under high temperature conditions.

Example 5 Effect of the buffer salt species on the stability of pharmaceutical formulations comprising GR1603 protein

Liquid formulations were prepared using the preparation method described in example 1 and according to the recipe composition shown in the following table, except that the histidine/histidine salt buffer pair was used instead of the acetic acid/acetate buffer pair and the pH of the formulation was adjusted.

TABLE 15 prescription composition of liquid formulations

The purity of each formulation sample was tested using storage conditions, sampling time points and test items similar to those described in example 1. The detection results are shown in the following table and fig. 6A to 6G.

TABLE 16 influence of buffer salt species on Tm values of pharmaceutical formulations comprising GR1603 protein

Formulation number	Tm1 value (. Degree. C.)	Tm2 value (. Degree. C.)
			S1	54.6	66.1
S14	54.5	Undetected
			S15	54.9	66.5
S16	54.7	66.4

Undetected ×: the DSF fluorescence signal derivative curve of the S14 formulation does not have the 2 nd peak, and Tm2 value cannot be calculated.

TABLE 17 influence of the buffer salt species on the content of related substances in pharmaceutical formulations comprising GR1603 protein

N/A: and not detected.

As can be seen from the above table and FIGS. 6A to 6G, there is no difference in the Tm value of GR1603 protein in the buffer system of acetate and histidine salts. Under the high temperature condition, the CEX-HPLC purity change trend of the GR1603 protein is not obviously different in an acetate buffer system; in the histidine salt buffer system, the rate of increase of protein aggregates was slower and there was no significant difference in CEX-HPLC purity trend of GR1603 protein, but in the histidine salt buffer system at pH5.8, GR1603 protein was unstable, the rate of decrease of the monomer peak area% was faster, the rate of decrease of non-reducing CE-SDS purity was faster, and the rate of decrease of reducing CE-SDS purity was faster.

Example 6 in-depth investigation of the effect of osmolality adjusting agent on the stability of pharmaceutical formulations comprising GR1603 protein

And taking a proper amount of GR1603 protein sample, replacing the ultrafiltration liquid into pure water, and diluting with water until the protein concentration is 62.5mg/ml, thus obtaining the GR1603 protein aqueous solution. A 5× formulation buffer stock (containing 5-fold concentrations of buffer salt and osmolality adjusting agent, pH approximately 5.2) was prepared for each formulation shown in table 18. Then, 1.2ml of the GR1603 protein aqueous solution, 0.3ml of the 5 Xpreparation buffer mother liquor and 3. Mu.L of the 10% polysorbate 80 aqueous solution were taken and mixed uniformly to prepare preparation samples of different osmotic pressure regulators, respectively.

TABLE 18 prescription composition of liquid formulations

The concentration of arginine hydrochloride is respectively reduced to 80 percent (24 mg/ml), 50 percent (15 mg/ml), 20 percent (6 mg/ml) and 0 percent by taking 30mg/ml of isotonic arginine hydrochloride as 100 percent arginine hydrochloride; the concentration of trehalose was reduced to 80% (68 mg/ml), 50% (42.5 mg/ml), 20% (17 mg/ml) and 0%, respectively, with isotonic 85mg/ml trehalose as 100% trehalose.

The purity of each formulation sample was tested using storage conditions, sampling time points and test items similar to those described in example 1. The detection results are shown in the following table and fig. 7A to 8E.

TABLE 19 influence of osmolality regulator on the content of related substances in pharmaceutical formulations comprising GR1603 protein

/>

As can be seen from the above table and fig. 7A to 8E, the GR1603 protein aggregation rate is increased when the arginine hydrochloride content is reduced to less than 50% (15 mg/ml) under high temperature conditions; and the rate of increase of the acidic peak decreases with increasing arginine hydrochloride concentration. Under high temperature conditions, arginine hydrochloride, proline and arginine acetate did not significantly differ in inhibiting the increase in the GR1603 protein multimer, and the increase in the acidic peak was rapid in the proline-containing formulation.

Example 7 Effect of Polysorbate 80 concentration on stability of pharmaceutical formulations containing GR1603 protein

Liquid formulations were prepared according to the formulation shown in the following table using the preparation method in example 1.

Table 20 prescription composition

/>

The "%" in Table 20 means weight/volume percent or% w/v. For example, 0.02% polysorbate 80 means that the amount of polysorbate 80 in the formulation is 0.2mg/mL.

Samples of the liquid formulations of polysorbate 80 having different concentrations in table 20 were subjected to shaking experiments and after shaking at a rate of 1500 times/mm for 1 day, turbidity (OD value at 350nm/550 nm) and SEC-HPLC polymer levels in each sample were detected in a cuvette with a 1cm optical path using a spectrophotometer. The results of the shaking experiments are shown in FIGS. 9A-9C. When the sample of the preparation does not contain polysorbate 80, the turbidity of the sample is obviously increased after shaking for 1 day, which indicates that the preparation containing the GR1603 protein is easily affected by mechanical force such as shaking and the like to generate turbidity. When polysorbate 80 was added at a concentration of 0.01% to a formulation containing GR1603 protein, the turbidity of the sample did not increase significantly after shaking for 1 day, which had a significant effect on inhibiting turbidity of the formulation.

Samples of liquid formulations having different concentrations of polysorbate 80 in table 20 were subjected to freeze thawing experiments, i.e., the samples were repeatedly freeze-thawed 5 times at-20 c to room temperature, and SEC-HPLC polymer levels were determined, the results of which are shown in fig. 10. The results show that repeated freeze thawing does not increase the GR1603 protein polymer, and the addition of polysorbate 80 to the formulation does not affect the damage of freeze thawing to the protein.

EXAMPLE 8 formulation development at high protein concentration

An appropriate amount of GR1603 protein sample was removed, the stock solution was ultrafiltered with 20mM acetate Buffer, then diluted 1:1 by volume with each prescription of 2 Xarginine hydrochloride Buffer (containing 20mM acetate and 2 times the concentration of arginine hydrochloride) and concentrated to a protein concentration of 167mg/ml, finally 10 Xtrehalose Buffer stock solution (containing 20mM acetate, 1 times the concentration of arginine hydrochloride and 10 times the concentration of trehalose) was added to each prescription at a 9:1 volume ratio, and 10% polysorbate 80 aqueous solution was added to give a final formulation of polysorbate 80 concentration of 0.2mg/ml.

TABLE 21 prescription composition of high protein concentration formulations

The viscosity of each formulation was determined and the purity of each formulation sample was measured using storage conditions, sampling time points and test items similar to those described in example 1. The detection results are shown in the following table and fig. 11A to 11G.

TABLE 22 viscosity of high protein concentration formulations

/>

TABLE 23 purity variation of high protein concentration formulations at high temperature (37 ℃ C.)

Table 22 shows that the viscosities of the GR1603 formulations of each formulation at 150mg/ml concentrations are from about 10 mPas to about 12.5 mPas, meeting the viscosity requirements for subcutaneous injection administration.

As can be seen from Table 23 and FIGS. 11A through 11G, each GR1603 formulation at 150mg/ml concentration had good stability at high temperature, increased SEC-HPLC polymer ratio by 1.4% to 1.9% at 8 weeks, increased CEX-HPLC acid peak by 20.9% to 25.2%, decreased non-reducing CE-SDS purity by 6.1% to 6.8%, and decreased reducing CE-SDS purity by 2.2% to 3.6%.

Claims

1. A pharmaceutical composition comprising about 5mg/ml to about 200mg/ml of a recombinant fully human anti-IFNAR 1 monoclonal antibody, an osmolality adjusting agent selected from the group consisting of trehalose and/or arginine, optionally a non-ionic surfactant, and the balance water, wherein the pharmaceutical composition has a pH of about 4.8 to about 6.0, wherein the monoclonal antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 sequences, and specifically binds human type I interferon receptor subunit 1 (IFNAR 1).

2. The pharmaceutical composition of claim 1, comprising the nonionic surfactant, wherein the nonionic surfactant is a polysorbate-type surfactant, such as polysorbate 20 and polysorbate 80.

3. The pharmaceutical composition of claim 2, wherein the non-ionic surfactant is present in an amount of about 0.05mg/ml to about 1.0mg/ml, about 0.1mg/ml to about 0.3mg/ml, about 0.2mg/ml to about 0.5mg/ml, about 0.3mg/ml to about 1.0mg/ml or about 0.5mg/ml to about 1.0mg/ml, e.g., about 0.2mg/ml, about 0.3mg/ml, about 0.34mg/ml, about 0.4mg/ml, about 0.5mg/ml, about 0.6mg/ml, about 0.7mg/ml, about 0.8mg/ml, about 0.9mg/ml or about 1.0mg/ml.

4. A pharmaceutical composition according to any one of claims 1-3, wherein the osmolality adjusting agent is a pharmaceutically acceptable salt of arginine, such as arginine hydrochloride, or a mixture of trehalose and a pharmaceutically acceptable salt of arginine, such as a mixture of trehalose and arginine hydrochloride.

5. The pharmaceutical composition of claim 4, wherein the amount of the pharmaceutically acceptable salt of arginine contained in the mixture of trehalose and pharmaceutically acceptable salt of arginine is about 50 to about 95 parts by weight, about 55 to about 90 parts by weight, about 60 to about 90 parts by weight, about 65 to about 85 parts by weight, about 70 to about 80 parts by weight, or about 75 parts by weight relative to 100 parts by weight of the pharmaceutically acceptable salt of arginine, and the amount of trehalose contained in the mixture of trehalose and pharmaceutically acceptable salt of arginine is about 5 to about 50 parts by weight, about 10 to about 45 parts by weight, about 10 to about 40 parts by weight, about 15 to about 35 parts by weight, about 20 to about 30 parts by weight, or about 25 parts by weight relative to 100 parts by weight of trehalose.

6. The pharmaceutical composition of any one of claims 1-5, further comprising a buffer salt, such as a phosphate/phosphate buffer pair, a histidine/histidine inorganic salt buffer pair, an acetic acid/acetate buffer pair, a citric acid/citrate buffer pair, preferably an acetic acid/acetate buffer pair, such as an acetic acid/sodium acetate buffer pair.

7. The pharmaceutical composition of claim 6, wherein the buffer salt is at a concentration of about 10mM to about 50mM, about 10mM to about 45mM, about 10mM to about 40mM, about 10mM to about 35mM, about 10mM to about 30mM, about 15mM to about 25mM, about 15mM to about 20mM, about 20mM to about 25mM, or about 20mM.

8. The pharmaceutical composition of any one of claims 1-7, wherein the pH of the pharmaceutical composition is about 5.0 to about 5.8, about 5.0 to about 5.6, about 5.0 to about 5.5, about 5.2 to about 5.6, or about 5.2 to about 5.5.

9. Use of a pharmaceutical composition according to any one of claims 1 to 8 in the manufacture of a medicament for the prophylaxis or treatment of a disease mediated by human IFNAR 1.

10. The use of claim 9, wherein the disease mediated by human IFNAR1 is an autoimmune disease, such as systemic lupus erythematosus, psoriasis, multiple sclerosis or rheumatoid arthritis.