Anti-blys antibodies, pharmaceutical compositions thereof, and uses thereof

Abstract

The present application provides anti-BLyS antibodies, pharmaceutical compositions thereof, and uses thereof. The anti-BLyS antibodies or antigen-binding fragments thereof of the present application comprise a heavy chain variable region and a light chain variable region, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 thereof are shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively, and the amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively. The anti-BLyS antibodies or antigen-binding fragments thereof of the present application have excellent binding ability to human BLyS protein and the ability to competitively inhibit the binding of human BLyS to its receptor human BR3. The pharmaceutical composition of the present application is a highly stable pharmaceutical composition containing anti-BLyS antibodies and a buffer, and can further contain at least one stabilizer and optionally a surfactant.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and relates to an anti-BLyS antibody, its pharmaceutical composition and its use. The anti-BLyS antibody has excellent binding ability to human BLyS protein and competitive inhibition of the binding of human BLyS to its receptor human BR3. Background Technology

[0002] Systemic lupus erythematosus (SLE) is a diffuse, progressive, autoimmune disease that affects multiple systems throughout the body, including the skin, joints, heart, lungs, kidneys, blood, and brain, often alternating between remission and relapse. SLE primarily affects people of African and Caribbean descent, Asians, and Hispanics, while Caucasians (Caucasians) are less affected. Because the initial symptoms of SLE are often subtle, the actual number of patients is likely much higher than currently estimated. Therefore, there is a significant objective need for improved diagnosis and treatment of SLE in clinical practice.

[0003] The causes of systemic lupus erythematosus (SLE) are complex and unclear, and it is not caused by a single factor. It may be related to multiple factors such as genetics, environment, sex hormones, and immunity. Currently, the internationally recognized cause of SLE is that at the cellular level, self-reactive B cells persist for too long in peripheral tissues, producing autoantigens and causing autoimmunity. Therefore, if the growth and proliferation of early B cells can be inhibited, SLE can be treated.

[0004] B lymphocyte stimulator (BLyS), also known as Tall-1 (TNF and Apol-related leukocyte-expressed ligand 1), BAFF (B cell activating factor belonging to the TNF family), and THANK (TNF homologues that activate apoptosis, NF-κB, and JNK), belongs to the tumor necrosis factor (TNF) family. It is a novel cytokine first discovered and cloned by Shu Hongbing et al. in 1999. As a co-stimulatory factor for B lymphocytes, BLyS can specifically stimulate B cell proliferation and differentiation in the presence of anti-IgM and IL-4, playing a crucial role in humoral immunity; its overexpression in vivo is closely related to autoimmune diseases.

[0005] In vitro experiments showed that after B cells were preactivated with IgM, BLyS could induce their proliferation and secretion of large amounts of IgM and IgA. However, this stimulatory effect was not significant for B cells in the resting state (3). Further studies showed that BLyS mainly acts on pre-B lymphocytes, immature B lymphocytes, and activated lymphocytes, but has no effect on plasma cells or lymphoplastic stem cells. Like most cytokines, BLyS mainly stimulates downstream signal transduction through B cell surface receptors. Multiple research groups have confirmed that the receptors that bind to BLyS are: B cell activator receptor (BR3, BLyS receptor 3, or BAFF-R), transmembrane activator-1 and calcium modulator and cyclophilin ligand-interactor (TACI), and B cell maturation antigen (BCMA). This specificity determines that BLyS is a good target for B cell antibody-mediated autoimmune diseases and lymphoma.

[0006] Therapeutic antibodies against BLyS have been shown in vivo and in vitro to effectively inhibit B cell growth and IgA and IgM secretion, thereby achieving the therapeutic effect of treating systemic lupus erythematosus (Edwards BM et al., The remarkable flexibility of the human antibody repertoire; isolation of over one thousand different antibodies to a single protein, BLyS. J Mol Biol. 2003 Nov 14; 334(1):103-18; Baker KP et al., Generation and characterization of LymphoStat-B, a human monooclonal antibody that antagonizes the bioactivities of B lymphocyte stimulator. Arthritis Rheum. 2003 Nov; 48(11):3253-65). Benlysta, an anti-BLyS antibody developed by Human Genome Research, has become the world's first novel drug for the treatment of lupus erythematosus in the past 60 years. Benlysta targets only BLyS-stimulated B cells, significantly reducing side effects compared to chemotherapy drugs, thus providing a safe and effective treatment option for patients with systemic lupus erythematosus (SLE). In recent years, research and clinical application of targeted therapies against BLyS have developed rapidly. Apart from Human Genome Research, other companies have used fusion proteins modified based on BLyS or its receptor. Genentech developed BR3-FC, Zymogenetics developed TACI-FC, and AMGEN developed peptide-FC. Compared to Benlysta, these drugs have lower specificity, weaker binding affinity, relatively lower efficacy, and higher toxicity. All three drugs have been discontinued or terminated in phase II clinical trials. Therefore, anti-BLyS antibody drugs are the effective approach for targeting this target. Summary of the Invention

[0007] The anti-BLyS antibody provided by this invention possesses excellent binding ability to human BLyS protein and the ability to competitively inhibit the binding of human BLyS to its receptor, human BR3. The pharmaceutical composition is a highly stable pharmaceutical composition containing the anti-BLyS antibody. In particular, this invention has discovered that the humanized anti-BLyS antibody exhibits unexpected characteristics, namely, high stability, in histidine buffer systems and combinations of L-arginine, L-arginine hydrochloride, or methionine.

[0008] This invention provides a humanized anti-BLyS antibody or its antigen-binding fragment, comprising a heavy chain variable region and a light chain variable region, wherein:

[0009] The amino acid sequences of the humanized anti-BLyS antibody or its antigen-binding fragment LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, respectively;

[0010] The amino acid sequences of the humanized anti-BLyS antibody or its antigen-binding fragment HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, respectively;

[0011] The light chain variable region of the humanized anti-BLyS antibody or its antigen-binding fragment is selected from any of the FR1 described in SEQ ID NO:7, 9 and 13, FR2 is the FR2 described in SEQ ID NO:7, FR3 is selected from any of the FR3 described in SEQ ID NO:7, 9 and 11, and FR4 is the FR4 described in SEQ ID NO:7.

[0012] The heavy chain variable region of the humanized anti-BLyS antibody or its antigen-binding fragment is selected from FR1 of any one of SEQ ID NO:8, 10, 12 and 15, FR2 of any one of SEQ ID NO:8, 10 and 15, FR3 of any one of SEQ ID NO:8, 12 and 14, and FR4 of SEQ ID NO:8 or 10; and

[0013] The humanized anti-BLyS antibody or its antigen-binding fragment does not include: a humanized anti-BLyS antibody or its antigen-binding fragment having the amino acid sequence shown in SEQ ID NO:7 in the light chain variable region and the amino acid sequence shown in SEQ ID NO:8 in the heavy chain variable region.

[0014] In some protocols, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment contains:

[0015] The light chain variable region FR1 is selected from any one of SEQ ID NO:7, 9, and 13, FR2 is the FR2 described in SEQ ID NO:7, FR3 is selected from any one of SEQ ID NO:7, 9, and 11, and FR4 is the FR4 described in SEQ ID NO:7; the heavy chain variable region FR1-FR4 are respectively FR1-FR4 of SEQ ID NO:15; or

[0016] The light chain variable region FR1 is selected from any one of SEQ ID NO:7, 9, and 13, FR2 is the FR2 described in SEQ ID NO:7, FR3 is selected from any one of SEQ ID NO:7, 9, and 11, and FR4 is the FR4 described in SEQ ID NO:7; the heavy chain variable region FR1-FR4 are respectively FR1-FR4 of SEQ ID NO:10; or

[0017] The light chain variable region FR1 is selected from any one of SEQ ID NO:7, 9, and 13, FR2 is the FR2 described in SEQ ID NO:7, FR3 is selected from any one of SEQ ID NO:7, 9, and 11, and FR4 is the FR4 described in SEQ ID NO:7; the heavy chain variable region FR1-FR4 are respectively FR1-FR4 of SEQ ID NO:12; or

[0018] The light chain variable region FR1 is the FR1 described in SEQ ID NO:11 or 13, FR2 is the FR2 described in SEQ ID NO:11, FR3 is the FR3 described in SEQ ID NO:11 or 13, and FR4 is the FR4 described in SEQ ID NO:11; the heavy chain variable region FR1-FR4 are FR1-FR4 of SEQ ID NO:14 respectively.

[0019] In some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13, and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15.

[0020] In some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region and the heavy chain variable region are selected from:

[0021] (1) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0022] (2) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0023] (3) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0024] (4) The light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 13, and the heavy chain variable region contains an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15;

[0025] Preferably, the light chain variable region and the heavy chain variable region are selected from:

[0026] (5) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 10; or

[0027] (6) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 12; or

[0028] (7) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 14; or

[0029] (8) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0030] (9) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0031] (10) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0032] (11) The light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 13, and the heavy chain variable region contains an amino acid sequence as shown in SEQ ID NO: 15.

[0033] The humanized anti-BLyS antibody of this invention further comprises a human light chain constant region and a heavy chain constant region, and the light chain variable region and the heavy chain variable region are respectively linked to the human light chain constant region and the heavy chain constant region. That is, the humanized anti-BLyS antibody contains a complete light chain and a complete heavy chain, wherein the complete light chain is formed by linking the light chain variable region contained in the anti-BLyS antibody with the human light chain constant region, and the complete heavy chain is formed by linking the heavy chain variable region contained in the anti-BLyS antibody with the human heavy chain constant region.

[0034] In some schemes, the human light chain constant region is the human light chain κ constant region.

[0035] In some schemes, the human heavy chain constant region is the human heavy chain Fc segment.

[0036] The human light chain κ constant region and the human heavy chain Fc fragment are both derived from healthy human B lymphocytes. Using genetic engineering techniques, overlap extension PCR was used to ligate the variable and constant regions, resulting in the complete humanized light and heavy chains of the anti-BLyS antibody.

[0037] In some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein the light chain comprises an amino acid sequence as shown in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20 or SEQ ID NO: 22, and the heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24.

[0038] In some formulations, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein:

[0039] (1) The light chain comprises the amino acid sequence shown in SEQ ID NO: 16, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24; or

[0040] (2) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24; or

[0041] (3) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, or SEQ ID NO: 24; or

[0042] (4) The light chain comprises an amino acid sequence as shown in SEQ ID NO: 22, and the heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24;

[0043] Preferably:

[0044] (5) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 19; or

[0045] (6) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 21; or

[0046] (7) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 23; or

[0047] (8) The light chain comprises the amino acid sequence shown in SEQ ID NO: 16, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0048] (9) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0049] (10) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0050] (11) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24.

[0051] In some embodiments, the antigen-binding fragment described in this invention is selected from Fab, Fab', Fab'-SH, Fv, scFv, F(ab')2, sdAb, or a double antibody.

[0052] In another aspect, the present invention provides a polynucleotide molecule selected from: a polynucleotide molecule encoding a humanized anti-BLyS antibody as described in any of the embodiments herein, or an antigen-binding fragment thereof, or a complementary sequence thereof.

[0053] In another aspect, the present invention provides an expression vector comprising a polynucleotide molecule as described herein, preferably a eukaryotic expression vector.

[0054] In another aspect, the present invention provides a host cell comprising a polynucleotide molecule or expression vector as described herein, preferably a eukaryotic cell, more preferably a mammalian cell.

[0055] In another aspect, the present invention provides a method for preparing a humanized anti-BLyS antibody or an antigen-binding fragment thereof as described in any of the embodiments herein, the method comprising culturing a host cell as described herein under conditions suitable for expression of the antibody or the antigen-binding fragment thereof, to express the antibody or the antigen-binding fragment thereof, and recovering the expressed antibody or the antigen-binding fragment thereof.

[0056] In another aspect, the present invention provides a pharmaceutical composition comprising a humanized anti-BLyS antibody or an antigen-binding fragment thereof as described in any of the embodiments herein, a polynucleotide as described herein, an expression vector or a host cell as described herein, and a pharmaceutically acceptable carrier or excipient.

[0057] In some embodiments, the present invention provides a pharmaceutical composition comprising: (1) a buffer solution; and (2) a humanized anti-BLyS antibody or an antigen-binding fragment thereof, said humanized anti-BLyS antibody or antigen-binding fragment thereof having amino acid sequences as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, respectively, LCDR1, LCDR2 and LCDR3, and amino acid sequences as shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, respectively, HCDR1, HCDR2 and HCDR3.

[0058] In some embodiments, the present invention provides a pharmaceutical composition comprising: (1) a buffer solution; and (2) a humanized anti-BLyS antibody or an antigen-binding fragment thereof, wherein the humanized anti-BLyS antibody or the antigen-binding fragment thereof is as described in any embodiment herein.

[0059] In some formulations, the concentration of the humanized anti-BLyS antibody or its antigen-binding fragment in the above-mentioned pharmaceutical composition is about 1-300 mg / mL, preferably about 10-300 mg / mL, more preferably about 20-280 mg / mL, more preferably about 30-150 mg / mL, more preferably about 80-120 mg / mL, and even more preferably about 180-220 mg / mL.

[0060] In some schemes, the buffer solution is selected from one or more of acetate buffer, citrate buffer, succinate buffer, phosphate buffer, and histidine buffer.

[0061] In some formulations, the concentration of the buffer solution is approximately 1-200 mM, preferably approximately 1-100 mM, preferably approximately 5-50 mM, preferably approximately 10-40 mM, and preferably approximately 20-40 mM.

[0062] In some embodiments, the pH of the buffer solution is approximately 5.0-6.5, preferably approximately 5.0-6.0, and even more preferably approximately 5.5-6.0.

[0063] In some formulations, the aforementioned pharmaceutical composition also includes a stabilizer.

[0064] In some embodiments, the stabilizer is selected from one or more of arginine, arginine hydrochloride, methionine, proline, glycine, sodium chloride, mannitol, sorbitol, sucrose, maltose, xylitol, and trehalose.

[0065] In some formulations, the stabilizer is selected from one or more of L-arginine, L-arginine hydrochloride, sucrose, methionine, and sodium chloride.

[0066] In some formulations, the concentration of the stabilizer is about 10 mM to 400 mM, preferably about 50 mM to 300 mM, more preferably about 100 mM to 300 mM, and even more preferably about 100 mM to 200 mM.

[0067] In some formulations, the stabilizer is a combination of approximately 10-30 mM L-arginine hydrochloride and approximately 50-200 mM sodium chloride; or the stabilizer is a combination of approximately 10-30 mM L-arginine and approximately 50-200 mM sodium chloride; or the stabilizer is a combination of approximately 10-30 mM L-arginine and approximately 80-220 mM sucrose; or the stabilizer is a combination of approximately 10-30 mM L-arginine hydrochloride and approximately 80-220 mM sucrose; or the stabilizer is a combination of approximately 30-90 mM methionine and approximately 50-200 mM sodium chloride; or the stabilizer is approximately 10-120 mM L-arginine hydrochloride. Salt; or the stabilizer is about 10-120 mM methionine; preferably, the stabilizer is a combination of about 10-30 mM L-arginine hydrochloride and about 80-120 mM sodium chloride; or the stabilizer is a combination of about 10-30 mM L-arginine and about 80-120 mM sodium chloride; or the stabilizer is a combination of about 50-70 mM methionine and about 80-120 mM sodium chloride; or the stabilizer is a combination of about 10-30 mM L-arginine and about 130-170 mM sucrose; or the stabilizer is a combination of about 10-30 mM L-arginine hydrochloride and about 130-170 mM sucrose.

[0068] In some embodiments, the pharmaceutical composition further includes a surfactant, preferably selected from one or more of polysorbate 80, polysorbate 20, and poloxamer 188.

[0069] In some formulations, the concentration of the surfactant, calculated by w / v, is approximately 0.001%-0.1%, preferably approximately 0.01%-0.1%, and most preferably approximately 0.01%-0.05%.

[0070] In some formulations, the above-mentioned pharmaceutical composition also includes water for injection.

[0071] In some embodiments, the pharmaceutical composition is a liquid formulation or a lyophilized formulation.

[0072] In some embodiments, the pharmaceutical composition is a liquid formulation.

[0073] In another aspect, the present invention provides an injection comprising the pharmaceutical composition described in any of the embodiments herein and a sodium chloride solution; preferably, the concentration of the sodium chloride solution is 0.85 to 0.9%; preferably, the concentration of the humanized anti-BLyS antibody in the injection is 3 to 100 mg / mL, more preferably about 3 to 60 mg / mL; preferably, the pH of the injection is 5.5 to 6.0.

[0074] In some embodiments, the pharmaceutical composition or injection is administered via subcutaneous injection.

[0075] In another aspect, the present invention provides the use of humanized anti-BLyS antibodies or antigen-binding fragments thereof as described in any of the embodiments herein, polynucleotide molecules described herein, expression vectors described herein, host cells described herein, pharmaceutical compositions described herein, or injections described herein in the preparation of medicaments for the prevention and / or treatment of diseases caused by excessive B cell proliferation.

[0076] In another aspect, the present invention provides a humanized anti-BLyS antibody or its antigen-binding fragment as described in any of the embodiments herein, a polynucleotide molecule described herein, an expression vector described herein, a host cell described herein, a pharmaceutical composition described herein, or an injection described herein, for the prevention and / or treatment of diseases caused by excessive B cell proliferation.

[0077] In another aspect, the present invention provides a method for preventing and / or treating diseases caused by excessive B cell proliferation, comprising administering to a subject in need a humanized anti-BLyS antibody or an antigen-binding fragment thereof as described in any of the embodiments herein, a polynucleotide molecule described herein, an expression vector described herein, a host cell described herein, a pharmaceutical composition described herein, or an injection described herein.

[0078] Preferably, the diseases caused by excessive B cell proliferation are systemic lupus erythematosus, rheumatoid arthritis, ankylosing spondylitis, or B-cell lymphoma.

[0079] In another aspect, the present invention provides a pharmaceutical combination comprising a humanized anti-BLyS antibody or an antigen-binding fragment thereof as described in any of the embodiments herein, a polynucleotide molecule described herein, an expression vector described herein, a host cell described herein, a pharmaceutical composition described herein, or an injectable formulation described herein, and one or more other therapeutic agents.

[0080] In another aspect, the present invention provides a kit comprising a humanized anti-BLyS antibody or an antigen-binding fragment thereof as described in any of the embodiments herein, a polynucleotide molecule as described herein, an expression vector as described herein, a host cell as described herein, a pharmaceutical composition as described herein, or an injectable as described herein, preferably further comprising a drug delivery device.

[0081] In another aspect, the present invention provides the use of histidine buffer and one or more stabilizers selected from L-arginine, L-arginine hydrochloride, methionine, and sodium chloride, and optionally a surfactant (preferably polysorbate 80), in improving the stability of pharmaceutical formulations of humanized anti-BLyS antibodies or antigen-binding fragments thereof, or in preparing pharmaceutical formulations of humanized anti-BLyS antibodies or antigen-binding fragments thereof with improved stability. Preferably, the histidine buffer, stabilizer, and surfactant, and the amounts thereof, are as described in any embodiment herein; the stability improvement is as described in any embodiment herein; and the humanized anti-BLyS antibody or antigen-binding fragment thereof is as described in any embodiment herein. Detailed Implementation

[0082] Definitions and Explanations

[0083] To facilitate understanding of this invention, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this invention pertains. It should be understood that this invention is not limited to specific methods, reagents, compounds, compositions, or biological systems, and variations thereof are certainly possible. It should also be understood that the terminology used in this application is for the purpose of describing specific embodiments only and is not intended to be limiting.

[0084] Unless otherwise expressly stated, the singular forms "a," "an," and "the" used in this specification and the appended claims include plural references. Thus, for example, reference to "a polypeptide" includes a combination of two or more polypeptides, etc.

[0085] The terms "pharmaceutical composition" or "formulation" refer to a mixture containing one or more antibodies described herein, along with other components such as physiologically pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertment of its biological activity.

[0086] The term "liquid formulation" refers to a formulation in a liquid state and is not intended to refer to a lyophilized formulation in a weighted suspension. The liquid formulation of the present invention is stable during storage, and its stability is independent of lyophilization (or other state-changing methods, such as spray drying).

[0087] The term "aqueous liquid formulation" refers to a liquid formulation that uses water as a solvent. In some formulations, aqueous liquid formulations are formulations that do not require lyophilization, spray drying, and / or freezing to maintain stability (e.g., chemical and / or physical stability and / or biological activity).

[0088] The term "excipient" refers to an agent that can be added to a formulation to provide desired properties (such as consistency, improved stability) and / or adjust osmotic pressure. Common examples of excipients include, but are not limited to, sugars, polyols, amino acids, surfactants, and polymers.

[0089] The term “about” as used in this application, when referring to measurable values ​​(such as quantity, duration, etc.), is intended to cover variations of ±20% or ±10% relative to a specific value, including ±5%, ±1%, and ±0.1%, because such variations are suitable for carrying out the disclosed methods.

[0090] The term "buffer solution pH of approximately 5.0-6.5" refers to a reagent that, through the action of its acid / base conjugate components, enables a solution containing the reagent to resist pH changes. The buffer solution used in the formulations of this invention may have a pH in the range of approximately 5.0 to approximately 6.5, or a pH in the range of approximately 5.5 to approximately 6.5, or a pH in the range of approximately 5.0 to approximately 6.0.

[0091] In this paper, examples of “buffer solutions” that control the pH within this range include succinic acid, succinates (e.g., sodium succinate), gluconic acid, histidine, histidine hydrochloride, methionine, citric acid, citrate, phosphoric acid, phosphate, citrate / phosphate, imidazole, acetic acid, acetate, combinations thereof, and other organic acid buffers.

[0092] "Histidine buffer" is a buffer containing histidine ions. Examples of histidine buffers include histidine and histidine salts, such as histidine hydrochloride, histidine acetate, histidine phosphate, and histidine sulfate, such as a histidine buffer containing histidine and histidine hydrochloride; the histidine buffer of the present invention also includes a histidine buffer containing histidine and acetate (such as sodium or potassium salt).

[0093] "Citrate buffer" is a buffer solution containing citrate ions. Examples of citrate buffer solutions include citrate-sodium citrate, citrate-potassium citrate, citrate-calcium citrate, citrate-magnesium citrate, etc. Preferably, the citrate buffer solution is citrate-sodium citrate buffer solution.

[0094] "Acetate buffer" is a buffer solution containing acetate ions. Examples of acetate buffer solutions include acetate-sodium acetate, acetate-potassium acetate, acetate-calcium acetate, acetate-magnesium acetate, etc. Preferably, the acetate buffer solution is acetate-sodium acetate buffer.

[0095] "Succinate buffer" is a buffer solution containing succinate ions. Examples of succinate buffer solutions include sodium succinate-sodium succinate, potassium succinate-potassium succinate, calcium succinate-calcium succinate, magnesium succinate-magnesium succinate, etc. Preferably, the succinate buffer solution is sodium succinate-sodium succinate buffer solution.

[0096] Phosphate buffer is a buffer solution containing phosphate ions. Examples of phosphate buffers include disodium hydrogen phosphate-sodium dihydrogen phosphate and dipotassium hydrogen phosphate-potassium dihydrogen phosphate. Preferably, the phosphate buffer is disodium hydrogen phosphate-sodium dihydrogen phosphate.

[0097] The term "stabilizer" refers to a pharmaceutically acceptable excipient that protects the active pharmaceutical ingredient and / or formulation from chemical and / or physical degradation during manufacturing, storage, and application. Stabilizers include, but are not limited to, sugars, amino acids, salts, polyols, and their metabolites as defined below, such as sodium chloride, calcium chloride, magnesium chloride, mannitol, sorbitol, sucrose, trehalose, methionine, L-arginine or its salts (e.g., L-arginine hydrochloride), glycine, alanine (α-alanine, β-alanine), betaine, leucine, lysine, glutamic acid, aspartic acid, proline, 4-hydroxyproline, sarcosine, γ-aminobutyric acid (GABA), opins, alanine, strombine, and trimethylamine N-oxide (TMAO), human serum albumin (hsa), bovine serum albumin (bsa), α-casein, globulin, α-lactalbumin, LDH, lysozyme, myoglobin, ovalbumin, and RNAase A. Some stabilizers, such as sodium chloride, calcium chloride, magnesium chloride, mannitol, sorbitol, and sucrose, can also control osmotic pressure. The stabilizers specifically used in this invention are selected from one or more of polyols, amino acids, salts, and sugars. Preferably, the sugars are sucrose and trehalose, and preferably, the polyol is mannitol. Preferably, the amino acids are L-arginine or its salts (such as L-arginine hydrochloride), methionine, glycine, and proline. Preferably, the stabilizer is sodium chloride, mannitol, sorbitol, sucrose, trehalose, methionine, L-arginine, L-arginine hydrochloride, glycine, proline, sodium chloride-L-arginine, sodium chloride-L-arginine hydrochloride, sodium chloride-methionine, sodium chloride-sorbitol, sodium chloride-mannitol, sodium chloride-sucrose, sodium chloride-trehalose, L-arginine hydrochloride-mannitol, L-arginine hydrochloride-sucrose, more preferably L-arginine hydrochloride, L-arginine, methionine, sodium chloride-L-arginine, sodium chloride-L-arginine hydrochloride, sodium chloride-methionine, more preferably sodium chloride-L-arginine, sodium chloride-L-arginine hydrochloride. In a particularly preferred embodiment, the stabilizer used in this invention is selected from one or more of methionine, L-arginine, L-arginine hydrochloride, and sodium chloride.

[0098] The term "surfactant" generally includes agents that protect proteins, such as antibodies, from air / solution interface-induced stress and solution / surface-induced stress to reduce antibody aggregation or minimize particulate formation in formulations. Exemplary surfactants include, but are not limited to, nonionic surfactants such as polyoxyethylene sorbitan fatty acid esters (e.g., polysorbate 20 and polysorbate 80), polyethylene-polypropylene copolymers, polyethylene-polypropylene glycol, polyoxyethylene-stearate, polyoxyethylene alkyl ethers, such as polyoxyethylene monolauryl ether, alkylphenyl polyoxyethylene ether (Triton-X), polyoxyethylene-polyoxypropylene copolymers (pluronic), and sodium dodecyl sulfate (SDS). In a particularly preferred embodiment, the surfactant used in this invention is polysorbate 80.

[0099] The term "isotonic" means that the preparation has an osmotic pressure that is essentially the same as that of human blood. Isotonic preparations generally have an osmotic pressure of approximately 250 to 350 mOsm. Isotonicity can be measured using a vapor pressure or freezing point depression osmoremeter.

[0100] The term "stable" formulation refers to a formulation in which the antibody substantially retains its physical and / or chemical stability and / or biological activity during the manufacturing process and / or storage. A pharmaceutical formulation may be considered stable even if the contained antibody fails to retain 100% of its chemical structure or biological function after a certain period of storage. In some cases, maintaining approximately 90%, 95%, 96%, 97%, 98%, or 99% of the antibody structure or function after a certain period of storage may also be considered "stable." Various analytical techniques for measuring protein stability are available in this field and are reviewed in *Peptide and Protein Drug Delivery* 247-301, edited by Vincent Lee, Marcel Dekker, Inc., New York, NY, Pubs. (1991), and Jones, A. (1993) Adv. Drug Delivery Rev. 10: 29-90 (both incorporated herein by reference).

[0101] After a formulation has been stored at a certain temperature for a certain period of time, its stability can be measured by determining the percentage of natural antibodies remaining (and other methods). Among other methods, the percentage of natural antibodies can be measured by size exclusion chromatography (e.g., size exclusion high-performance liquid chromatography [SEC-HPLC]), where "natural" refers to non-aggregated and non-degraded antibodies. In some formulations, protein stability is determined by the percentage of monomeric proteins in a solution with a low percentage of degradation (e.g., fragmentation) and / or aggregated proteins. In some formulations, the formulation can be stored stably at room temperature, about 25–30°C, or 40°C for at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer, with a maximum of about 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of aggregated antibodies.

[0102] Stability can be measured (and by other methods) by determining the percentage of antibody (“acidic form”) that migrates in the more acidic fraction of the main antibody fraction (“major charged form”) during ion exchange, wherein stability is inversely proportional to the percentage of acidic antibody. Among other methods, the percentage of “acidified” antibody can be measured by ion exchange chromatography (e.g., cation exchange high-performance liquid chromatography [CEX-HPLC]). In some embodiments, acceptable stability means that after storage at a certain temperature for a certain period of time, the detectable acidic antibody form does not exceed approximately 49%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1%. The storage period prior to stability measurement may be at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. When assessing stability, the permissible storage temperature for pharmaceutical preparations may be approximately -80°C.

[0103] Storage at any temperature in the range of approximately 45°C, such as at approximately -80°C, approximately -30°C, approximately -20°C, approximately 0°C, approximately 2-8°C, approximately 5°C, approximately 25°C, or approximately 40°C.

[0104] If an antibody does not show substantially no signs of aggregation, precipitation, and / or denaturation upon visual inspection of color and / or clarity, or upon measurement by UV light scattering or size exclusion chromatography, then the antibody “maintains its physical stability” in the pharmaceutical composition. Aggregation is the process by which individual molecules or complexes covalently or nonvalently associate to form aggregates. Aggregation can proceed to the extent that visible precipitates are formed.

[0105] The stability of a formulation, such as its physical stability, can be assessed using methods known in the art, including measuring the apparent extinction (absorbance or optical density) of the sample. Such extinction measurements are related to the turbidity of the formulation. The turbidity of a formulation is partly an inherent property of proteins dissolved in solution and is typically measured by turbidimetry and quantified in turbidimetric units (NTU).

[0106] The turbidity level, which varies with the concentration of one or more components in the solution (e.g., protein and / or salt concentration), is also referred to as the “emulsification” or “emulsive appearance” of the formulation. The turbidity level can be calculated by referring to a standard curve generated using a suspension of known turbidity. Reference standards used to determine the turbidity level of a pharmaceutical composition can be based on the European Pharmacopoeia (European Pharmacopoeia, 4th edition, “Directorate for the Quality of Medicine of the Council of Europe” (EDQM), Strasbourg, France). According to the European Pharmacopoeia, a clear solution is defined as a solution with a turbidity less than or equal to that of a reference suspension having a turbidity of approximately 3 according to the European Pharmacopoeia. Turbidimetric turbidity measurements can detect Rayleigh scattering in the absence of association or non-ideal effects, which typically varies linearly with concentration. Other methods for assessing physical stability are well known in the art.

[0107] An antibody is considered to retain its biological activity as defined below if its chemical stability at a given time point is sufficient to ensure that the antibody retains its chemical stability in the pharmaceutical composition. Chemical stability can be assessed, for example, by detecting or quantifying chemical changes in the antibody. Chemical changes can include size changes (e.g., shortening), which can be assessed using, for example, size exclusion chromatography, SDS-PAGE, and / or matrix-assisted laser desorption / ionization / time-of-flight mass spectrometry (MALDI / TOF MS). Other types of chemical changes include charge changes (e.g., occurring as a result of deamidation or oxidation), which can be assessed, for example, by ion exchange chromatography.

[0108] If the antibody in the pharmaceutical composition is biologically active for its intended purpose, then the antibody “retains its biological activity” in the pharmaceutical composition. For example, if, after storage at temperatures such as 5°C, 25°C, and 45°C for a certain period of time (e.g., 1 to 12 months), the affinity of the humanized monoclonal antibody contained in the formulation for binding to COVID-19 is at least 90%, 95%, or greater than the antibody binding affinity before the storage, then the formulation of the present invention can be considered stable. Binding affinity can also be determined using, for example, ELISA or plasmon resonance techniques.

[0109] In the context of this invention, in a pharmacological sense, a "therapeutic effective amount" or "effective amount" of an antibody refers to an amount effective in preventing, treating, or alleviating symptoms of an impairment for which the antibody can be effectively treated. In this invention, a "therapeutic effective amount" or "therapeutic effective dose" of a drug is any amount of the drug that, when used alone or in combination with another therapeutic agent, protects a subject from disease onset or promotes disease regression, said disease regression being demonstrated by a reduction in the severity of disease symptoms, an increase in the frequency and duration of asymptomatic periods of the disease, or prevention of injury or disability caused by disease suffering. The ability of a drug to promote disease regression can be evaluated using a variety of methods known to those skilled in the art, such as in human subjects during clinical trials, in animal model systems predicting human efficacy, or by determining the activity of the drug in an in vitro assay. A therapeutically effective amount of a drug includes a "preventatively effective amount," which is any amount of the drug that, when given alone or in combination with other therapeutic agents to a subject at risk of disease or a subject experiencing disease relapse, inhibits the development or recurrence of the disease.

[0110] The terms "subject" or "patient" are intended to include mammalian organisms. Examples of subjects / patients include humans and non-human mammals, such as non-human primates, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non-human animals. In a particular embodiment of the invention, the subject is a human.

[0111] The terms “administration,” “giving,” and “treatment” refer to the introduction of a composition containing a therapeutic agent into a subject using any of the various methods or delivery systems known to those skilled in the art. Routes of administration of humanized anti-BLyS antibodies or their antigen-binding fragments include intravenous, intramuscular, subcutaneous, peritoneal, spinal, or other parenteral routes, such as injection or infusion. “Parenteral administration” refers to administration by injection other than enteral or local administration, including but not limited to intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intra-tracheal, intracapsular, intra-articular, intra-articular, intra-articular, subcapsular, subarachnoid, intraspinal, intradural, and intrasternal injections and infusions, as well as intracorporeal electroporation.

[0112] Antibody

[0113] The term "antibody" as used herein should be understood to include both the complete antibody molecule and its antigen-binding fragment. The term "antigen-binding portion" or "antigen-binding fragment" (or simply "antibody portion" or "antibody fragment") of an antibody, as used herein, refers to one or more fragments of the antibody that retain the ability to specifically bind to BLyS.

[0114] As used herein, the term "full-length antibody" or "complete antibody molecule" refers to an immunoglobulin molecule containing four peptide chains: two heavy (H) chains (approximately 50-70 kDa in full length) and two light (L) chains (approximately 25 kDa in full length) linked together by disulfide bonds. Each heavy chain (abbreviated as HC in this document) consists of a heavy chain variable region (abbreviated as VH in this document) and a heavy chain constant region (abbreviated as CH in this document). The heavy chain constant region consists of three domains: CH1, CH2, and CH3. Each light chain (abbreviated as LC in this document) consists of a light chain variable region (abbreviated as VL in this document) and a light chain constant region. The light chain constant region consists of one domain: CL. The VH and VL regions can be further subdivided into highly variable complementarity-determining regions (CDRs) and regions separated by more conserved regions called framework regions (FRs). Each VH or VL region consists of three CDRs and four FRs arranged in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4, from the amino terminus to the carboxyl terminus. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant regions of the antibody mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[0115] When used in this document, the term "CDR" refers to the complementarity-determining region within the antibody variable sequence. Three CDRs exist in each variable region of the heavy and light chains, which are named HCDR1, HCDR2, and HCDR3, or LCDR1, LCDR2, and LCDR3, for each heavy and light chain variable region. The precise boundaries of these CDRs are defined differently depending on the system.

[0116] The precise amino acid sequence boundary of the variable region CDR of the antibody of the present invention can be determined using any of many well-known schemes, including the Kabat scheme described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD” (“Kabat” numbering scheme) and the IMGT scheme described by Lefranc M.-P. et al. (1999 Nucleic Acids Research, 27, 209-212).

[0117] As used herein, “antigen-binding fragment” includes fragments of antibodies or derivatives thereof, typically comprising at least one fragment of the antigen-binding region or variable region (e.g., one or more CDRs) of the parent antibody, which retains at least some of the binding specificity of the parent antibody. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; biantibodies; linear antibodies; single-chain antibody molecules, such as sc-Fv; nanobodies formed from antibody fragments; and multispecific antibodies. When the binding activity of an antibody is expressed on a molar concentration basis, the binding fragment or derivative thereof typically retains at least 10% of the antigen-binding activity of the parent antibody. Preferably, the binding fragment or derivative thereof retains at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or higher of the antigen-binding affinity of the parent antibody. It is also anticipated that the antigen-binding fragment of an antibody may include conserved or non-conserved amino acid substitutions (referred to as “conserved variants” or “functionally conserved variants” of the antibody) that do not significantly alter its biological activity.

[0118] When referring to ligand / receptor, antibody / antigen, or other binding pairs, "specific" binding means the determination of the presence of said protein within a heterogeneous population of proteins and / or other biological reagents. For example, the binding reaction of the monoclonal antibody of this invention to the BLyS protein. Therefore, under specified conditions, a particular ligand / antigen binds to a specific receptor / antibody and does not bind in significant amounts to other proteins present in the sample.

[0119] The humanized monoclonal antibodies or antigen-binding fragments thereof described herein include any of the anti-BLyS antibodies described in application number CN201210160474.3, the entire disclosure of which is incorporated herein by reference. In some embodiments, the CDR sequence of the antibody used in the methods and compositions of the present invention includes the CDR sequence of the antibody BLyS-IV described in CN201210160474.3. In some embodiments, the non-limiting, exemplary antibodies used in the examples herein are selected from the humanized anti-BLyS antibody BLyS-IV described in CN201210160474.3.

[0120] In some embodiments, the non-limiting, exemplary humanized anti-BLyS antibody used in the examples herein has a light chain amino acid sequence as shown in SEQ ID NO:18 and a heavy chain amino acid sequence as shown in SEQ ID NO:19; this humanized anti-BLyS antibody is named H482L472.

[0121] In some embodiments, this invention provides a humanized anti-BLyS antibody or its antigen-binding fragment thereof, which can bind to B lymphocyte stimulating factor and inhibit the binding of B lymphocyte stimulating factor to its receptor BR3-Fc, exhibiting excellent binding ability to human BLyS protein and competitive inhibition of the binding of human BLyS to its receptor human BR3. The humanized anti-BLyS antibody or its antigen-binding fragment provided by this invention can be obtained by constructing an expression vector according to the methods shown in Examples 7 and 8 of application number CN201210160474.3, transiently transfecting it using 293F cells, and undergoing one-step affinity purification in a 100 mL system.

[0122] In some embodiments, the present invention provides a humanized anti-BLyS antibody or its antigen-binding fragment thereof, wherein the amino acid sequences of LCDR1, LCDR2, and LCDR3 of the humanized anti-BLyS antibody or its antigen-binding fragment are shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; and the amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. The light chain variable region FR1 is selected from any one of SEQ ID NO:7, 9, and 13; FR2 is the FR2 described in SEQ ID NO:7; FR3 is selected from any one of SEQ ID NO:7, 9, and 11; and FR4 is the FR4 described in SEQ ID NO:7. The heavy chain variable region FR1 is selected from any one of SEQ ID NO:8, 10, 12, and 15; FR2 is selected from any one of SEQ ID NO:8, 10, and 15; FR3 is selected from any one of SEQ ID NO:8, 12, and 14; and FR4 is the FR4 described in SEQ ID NO:7. FR4 as described in NO:8 or 10.

[0123] In some embodiments, the humanized anti-BLyS antibody or its antigen-binding fragment described in this invention does not include a humanized anti-BLyS antibody or its antigen-binding fragment having the amino acid sequence shown in SEQ ID NO:7 for the light chain variable region and the amino acid sequence shown in SEQ ID NO:8 for the heavy chain variable region. In some embodiments, the humanized anti-BLyS antibody or its antigen-binding fragment described in this invention does not include a humanized anti-BLyS antibody or its antigen-binding fragment having the amino acid sequence shown in SEQ ID NO:16 for the light chain and the amino acid sequence shown in SEQ ID NO:17 for the heavy chain.

[0124] In some embodiments, in the humanized anti-BLyS antibody or its antigen-binding fragment of the present invention, the amino acid sequences of LCDR1, LCDR2, and LCDR3 are as shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; the amino acid sequences of HCDR1, HCDR2, and HCDR3 are as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively; FR1 of the light chain variable region is selected from any of the FR1 described in SEQ ID NO:7, 9, and 13; FR2 is the FR2 described in SEQ ID NO:7; FR3 is selected from any of the FR3 described in SEQ ID NO:7, 9, and 11; and FR4 is the FR4 described in SEQ ID NO:7; FR1-FR4 of the heavy chain variable region are FR1-FR4 of SEQ ID NO:15, respectively.

[0125] In some embodiments, in the humanized anti-BLyS antibody or its antigen-binding fragment of the present invention, the amino acid sequences of LCDR1, LCDR2, and LCDR3 are as shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; the amino acid sequences of HCDR1, HCDR2, and HCDR3 are as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively; FR1 of the light chain variable region is selected from any of the FR1 described in SEQ ID NO:7, 9, and 13; FR2 is the FR2 described in SEQ ID NO:7; FR3 is selected from any of the FR3 described in SEQ ID NO:7, 9, and 11; and FR4 is the FR4 described in SEQ ID NO:7; FR1-FR4 of the heavy chain variable region are FR1-FR4 of SEQ ID NO:10, respectively.

[0126] In some embodiments, in the humanized anti-BLyS antibody or its antigen-binding fragment of the present invention, the amino acid sequences of LCDR1, LCDR2, and LCDR3 are as shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; the amino acid sequences of HCDR1, HCDR2, and HCDR3 are as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively; FR1 of the light chain variable region is selected from any of the FR1 described in SEQ ID NO:7, 9, and 13; FR2 is the FR2 described in SEQ ID NO:7; FR3 is selected from any of the FR3 described in SEQ ID NO:7, 9, and 11; and FR4 is the FR4 described in SEQ ID NO:7; FR1-FR4 of the heavy chain variable region are FR1-FR4 of SEQ ID NO:12, respectively.

[0127] In some embodiments, in the humanized anti-BLyS antibody or its antigen-binding fragment of the present invention, the amino acid sequences of LCDR1, LCDR2, and LCDR3 are as shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; the amino acid sequences of HCDR1, HCDR2, and HCDR3 are as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively; the FR1 of the light chain variable region is the FR1 described in SEQ ID NO:11 or 13, the FR2 is the FR2 described in SEQ ID NO:11, the FR3 is the FR3 described in SEQ ID NO:11 or 13, and the FR4 is the FR4 described in SEQ ID NO:11; the FR1-FR4 of the heavy chain variable region are FR1-FR4 of SEQ ID NO:14, respectively.

[0128] In some embodiments, the present invention provides a humanized anti-BLyS antibody or an antigen-binding fragment thereof, comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13, and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15.

[0129] In some formulations, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment includes a light chain variable region and a heavy chain variable region, wherein:

[0130] (1) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0131] (2) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0132] (3) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0133] (4) The light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 13, and the heavy chain variable region contains an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15.

[0134] Preferably, the light chain variable region and the heavy chain variable region are selected from one of the following groups:

[0135] (5) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 10; or

[0136] (6) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 12; or

[0137] (7) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 14; or

[0138] (8) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0139] (9) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0140] (10) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0141] (11) The light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 13, and the heavy chain variable region contains an amino acid sequence as shown in SEQ ID NO: 15.

[0142] In some formulations, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment includes a light chain variable region and a heavy chain variable region, wherein:

[0143] (a) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13;

[0144] (b) The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 10, and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 7, 9, 11 or 13;

[0145] (c) The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 12, and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 7, 9, 11 or 13;

[0146] (d) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 14, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11 or 13; or

[0147] (e) The heavy chain variable region contains an amino acid sequence as shown in SEQ ID NO: 15, and the light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 7, 9, 11 or 13.

[0148] The humanized anti-BLyS antibody of this invention further comprises a human light chain constant region and a heavy chain constant region, and the light chain variable region and the heavy chain variable region are respectively linked to the human light chain constant region and the heavy chain constant region. That is, the humanized anti-BLyS antibody contains a complete light chain and a complete heavy chain, wherein the complete light chain is formed by linking the light chain variable region contained in the anti-BLyS antibody with the human light chain constant region, and the complete heavy chain is formed by linking the heavy chain variable region contained in the anti-BLyS antibody with the human heavy chain constant region.

[0149] In some schemes, the human light chain constant region is the human light chain κ constant region.

[0150] In some schemes, the human heavy chain constant region is the human heavy chain Fc segment.

[0151] The human light chain κ constant region and the human heavy chain Fc fragment are both derived from healthy human B lymphocytes. Using genetic engineering techniques, overlap extension PCR was used to ligate the variable and constant regions, resulting in the complete humanized light and heavy chains of the anti-BLyS antibody.

[0152] In some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein the light chain comprises an amino acid sequence as shown in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20 or SEQ ID NO: 22, and the heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24.

[0153] In some formulations, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein:

[0154] (1) The light chain comprises the amino acid sequence shown in SEQ ID NO: 16, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24; or

[0155] (2) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24; or

[0156] (3) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, or SEQ ID NO: 24; or

[0157] (4) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, or SEQ ID NO: 24; or

[0158] Preferably, the light chain and the heavy chain are selected from one of the following groups:

[0159] (5) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 19; or

[0160] (6) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 21; or

[0161] (7) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 23; or

[0162] (8) The light chain comprises the amino acid sequence shown in SEQ ID NO: 16, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0163] (9) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0164] (10) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0165] (11) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24.

[0166] In some formulations, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein:

[0167] (a) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, and the light chain comprises the amino acid sequence shown in SEQ ID NO: 22;

[0168] (b) The heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 19, and the light chain comprises an amino acid sequence as shown in SEQ ID NO: 16, 18, 20 or 22;

[0169] (c) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 21, and the light chain comprises the amino acid sequence shown in SEQ ID NO: 16, 18, 20 or 22;

[0170] (d) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 23, and the light chain comprises the amino acid sequence shown in SEQ ID NO: 20 or 22; or

[0171] (e) The heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 24, and the light chain comprises an amino acid sequence as shown in SEQ ID NO: 16, 18, 20 or 22.

[0172] Pharmaceutical preparations

[0173] The pharmaceutical composition described in this invention is a highly stable pharmaceutical composition containing an anti-BLyS antibody. In particular, this invention has discovered that the anti-BLyS antibody exhibits unexpected characteristics, namely, high stability, in histidine buffer systems and combinations of L-arginine, L-arginine hydrochloride, or methionine.

[0174] The present invention provides a pharmaceutical composition comprising the humanized anti-BLyS antibody or its antigen-binding fragment described herein, the polynucleotide described herein, the expression vector or host cell described herein, and a pharmaceutically acceptable carrier or excipient.

[0175] In some embodiments, the present invention provides a pharmaceutical composition comprising: (1) a buffer solution; and (2) a humanized anti-BLyS antibody or an antigen-binding fragment thereof, said humanized anti-BLyS antibody or antigen-binding fragment thereof having amino acid sequences as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, respectively, LCDR1, LCDR2 and LCDR3, and amino acid sequences as shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, respectively, HCDR1, HCDR2 and HCDR3.

[0176] Preferably, the humanized anti-BLyS antibody or its antigen-binding fragment in the pharmaceutical composition of the present invention is as described in any embodiment of the "Antibody" section of this application; or the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 7, and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 8; or the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein the light chain comprises an amino acid sequence as shown in SEQ ID NO: 16, and the heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 17.

[0177] For example, in some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13, and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15.

[0178] In some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein:

[0179] (1) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0180] (2) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0181] (3) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0182] (4) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; or

[0183] Preferably, the light chain variable region and the heavy chain variable region are selected from one of the following groups:

[0184] (5) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 10; or

[0185] (6) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 12; or

[0186] (7) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 14; or

[0187] (8) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0188] (9) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 9, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0189] (10) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 11, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0190] (11) The light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 15; or

[0191] (12) The light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 7, and the heavy chain variable region contains an amino acid sequence as shown in SEQ ID NO: 8.

[0192] The humanized anti-BLyS antibody of this invention further comprises a human light chain constant region and a heavy chain constant region, and the light chain variable region and the heavy chain variable region are respectively linked to the human light chain constant region and the heavy chain constant region. That is, the humanized anti-BLyS antibody contains a complete light chain and a complete heavy chain, wherein the complete light chain is formed by linking the light chain variable region contained in the anti-BLyS antibody with the human light chain constant region, and the complete heavy chain is formed by linking the heavy chain variable region contained in the anti-BLyS antibody with the human heavy chain constant region.

[0193] In some schemes, the human light chain constant region is the human light chain κ constant region.

[0194] In some schemes, the human heavy chain constant region is the human heavy chain Fc segment.

[0195] The human light chain κ constant region and the human heavy chain Fc fragment are both derived from healthy human B lymphocytes. Using genetic engineering techniques, overlap extension PCR was used to ligate the variable and constant regions, resulting in the complete humanized light and heavy chains of the anti-BLyS antibody.

[0196] In some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein the light chain comprises an amino acid sequence as shown in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20 or SEQ ID NO: 22, and the heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24.

[0197] In some embodiments, the aforementioned humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain and a heavy chain, wherein:

[0198] (1) The light chain comprises the amino acid sequence shown in SEQ ID NO: 16, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24; or

[0199] (2) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23 or SEQ ID NO: 24; or

[0200] (3) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, or SEQ ID NO: 24; or

[0201] (4) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, or SEQ ID NO: 24; or

[0202] Preferably, the light chain and the heavy chain are selected from one of the following groups:

[0203] (5) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 19; or

[0204] (6) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 21; or

[0205] (7) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 23; or

[0206] (8) The light chain comprises the amino acid sequence shown in SEQ ID NO: 16, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0207] (9) The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0208] (10) The light chain comprises the amino acid sequence shown in SEQ ID NO: 20, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0209] (11) The light chain comprises the amino acid sequence shown in SEQ ID NO: 22, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 24; or

[0210] (12) The light chain comprises the amino acid sequence shown in SEQ ID NO: 16, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 17.

[0211] In some embodiments, the antigen-binding fragment described in this invention is selected from Fab, Fab', Fab'-SH, Fv, scFv, F(ab')2, sdAb, or a double antibody.

[0212] In some formulations, the concentration of the humanized anti-BLyS antibody or its antigen-binding fragment in the pharmaceutical composition is about 1-300 mg / mL, preferably about 10-300 mg / mL, more preferably about 20-280 mg / mL, even more preferably about 30-150 mg / mL, even more preferably about 80-120 mg / mL, and even more preferably about 180-220 mg / mL; more preferably, the concentration of the above-mentioned humanized monoclonal antibody or its antigen-binding fragment is about 5 mg / mL, 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 60 mg / mL, 70 mg / mL, 80 mg / mL, or 90 mg / mL. L, 100mg / mL, 110mg / mL, 120mg / mL, 130mg / mL, 140mg / mL, 150mg / mL, 160mg / mL, 170mg / mL, 180mg / mL or 200mg / mL, preferably about 40mg / mL, 60mg / mL, 70mg / mL, 80mg / mL, 90mg / mL , 95mg / mL, 100mg / mL, 105mg / mL, 110mg / mL, 120mg / mL, 150mg / mL, 160mg / mL, 170mg / mL, 180mg / mL, 190mg / mL, 195mg / mL, 200mg / mL, 205mg / mL, 210mg / mL, 220mg / mL.

[0213] In some schemes, the buffer solution is selected from one or more of acetate buffer, citrate buffer, succinate buffer, phosphate buffer, and histidine buffer.

[0214] In some embodiments, the concentration of the buffer solution is approximately 1-200 mM, preferably approximately 1-100 mM, preferably approximately 5-50 mM, preferably approximately 10-40 mM, and preferably approximately 20-40 mM; non-limiting examples of the buffer solution concentration are approximately 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, or 180 mM, or any two values ​​within these ranges as endpoints, preferably 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, or 35 mM.

[0215] In some embodiments, the pH of the buffer solution is about 5.0-6.5, preferably about 5.0-6.0, and even more preferably about 5.5-6.0. Non-limiting examples of the pH of the buffer solution are about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, and preferably about 5.5, 5.8 or 6.0.

[0216] In some embodiments, the buffer solution is a histidine buffer solution. Preferably, the histidine buffer solution is selected from histidine-hydrochloride buffer solution or histidine-acetate buffer solution, with histidine-hydrochloride buffer solution being the most preferred.

[0217] In some formulations, the histidine-hydrochloride buffer is prepared from histidine and histidine hydrochloride, preferably L-histidine and L-histidine monohydrochloride. In some formulations, the histidine buffer is prepared from 5-40 mM L-histidine and 5-40 mM L-histidine monohydrochloride. In some formulations, the histidine buffer is prepared from histidine and histidine hydrochloride in a molar ratio of 1:1 to 1:4. In some formulations, the histidine buffer is prepared from histidine and histidine hydrochloride in a molar ratio of 1:2 to 1:2.5. In some formulations, the histidine buffer is prepared from histidine and histidine hydrochloride in a molar ratio of 1:2.3. In some formulations, the histidine buffer is a pH 5.8 histidine buffer prepared from about 10.0-11.0 mM L-histidine and about 24.0-25.0 mM L-histidine monohydrochloride. In some protocols, the preparation method for a 35mM histidine buffer at pH 5.8 involves first preparing 35mM L-histidine and L-histidine monohydrochloride solutions separately, and then mixing them to a pH of 5.8. In other protocols, the histidine buffer is a pH 5.8 histidine buffer prepared from approximately 10.0mM histidine and approximately 25.0mM histidine hydrochloride.

[0218] In some embodiments, the buffer solution is an acetate buffer solution. Preferably, the acetate buffer solution is an acetate-sodium acetate buffer solution or an acetate-potassium acetate buffer solution, with acetate-sodium acetate buffer solution being the most preferred.

[0219] In some embodiments, the buffer solution is a citrate buffer solution, and preferably, the citrate buffer solution is a citrate-sodium citrate buffer solution.

[0220] In some embodiments, the buffer solution is a succinate buffer solution, and preferably, the succinate buffer solution is a succinate-sodium succinate buffer solution.

[0221] In some schemes, the buffer solution is a phosphate buffer, preferably a sodium dihydrogen phosphate-disodium hydrogen phosphate buffer or a potassium dihydrogen phosphate-disodium hydrogen phosphate buffer.

[0222] In some formulations, the aforementioned pharmaceutical composition also includes a stabilizer.

[0223] In some embodiments, the stabilizer is selected from one or more of arginine, arginine hydrochloride, methionine, proline, glycine, sodium chloride, mannitol, sorbitol, sucrose, maltose, xylitol, and trehalose.

[0224] In some formulations, the stabilizer is selected from one or more of L-arginine, L-arginine hydrochloride, sucrose, methionine, and sodium chloride.

[0225] In some formulations, the concentration of the stabilizer is about 10 mM to 400 mM, preferably about 50 mM to 300 mM, more preferably about 100 mM to 300 mM, and even more preferably about 100 mM to 200 mM.

[0226] In some formulations, the stabilizer is a combination of approximately 10-30 mM L-arginine hydrochloride and approximately 50-200 mM sodium chloride; or the stabilizer is a combination of approximately 10-30 mM L-arginine and approximately 50-200 mM sodium chloride; or the stabilizer is a combination of approximately 10-30 mM L-arginine and approximately 80-220 mM sucrose; or the stabilizer is a combination of approximately 10-30 mM L-arginine hydrochloride and approximately 80-220 mM sucrose; or the stabilizer is a combination of approximately 30-90 mM methionine and approximately 50-200 mM sodium chloride; or the stabilizer is approximately 10-120 mM L-arginine hydrochloride. Salt; or the stabilizer is about 10-120 mM methionine; preferably, the stabilizer is a combination of about 10-30 mM L-arginine hydrochloride and about 80-120 mM sodium chloride; or the stabilizer is a combination of about 10-30 mM L-arginine and about 80-120 mM sodium chloride; or the stabilizer is a combination of about 50-70 mM methionine and about 80-120 mM sodium chloride; or the stabilizer is a combination of about 10-30 mM L-arginine and about 130-170 mM sucrose; or the stabilizer is a combination of about 10-30 mM L-arginine hydrochloride and about 130-170 mM sucrose.

[0227] In some embodiments, the stabilizer is composed of sodium chloride at a concentration of about 50-200 mM and L-arginine at a concentration of about 10-30 mM; preferably, the stabilizer is composed of sodium chloride at a concentration of about 80-120 mM and L-arginine at a concentration of about 10-30 mM; non-limiting examples of the stabilizer are composed of sodium chloride at concentrations of about 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM or 130 mM and L-arginine at concentrations of about 10 mM, 15 mM, 20 mM, 25 mM, or 30 mM.

[0228] In some embodiments, the stabilizer is composed of sodium chloride at a concentration of about 50-200 mM and L-arginine hydrochloride at a concentration of about 10-30 mM; preferably, the stabilizer is composed of sodium chloride at a concentration of about 80-120 mM and L-arginine hydrochloride at a concentration of about 10-30 mM; non-limiting examples of the stabilizer are composed of sodium chloride at concentrations of about 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM or 130 mM and L-arginine hydrochloride at concentrations of about 10 mM, 15 mM, 20 mM, 25 mM, or 30 mM.

[0229] In some embodiments, the stabilizer is composed of sodium chloride at a concentration of about 50-200 mM and methionine at a concentration of about 30-90 mM; preferably, the stabilizer is composed of sodium chloride at a concentration of about 80-120 mM and methionine at a concentration of about 50-70 mM; non-limiting embodiments of the stabilizer are composed of sodium chloride at concentrations of about 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM or 130 mM and methionine at concentrations of about 30 mM, 40 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM or 80 mM.

[0230] In some embodiments, the stabilizer is composed of L-arginine hydrochloride at a concentration of about 10-30 mM and sucrose at a concentration of about 80-220 mM; preferably, the stabilizer is composed of sucrose at a concentration of about 130-170 mM and L-arginine hydrochloride at a concentration of about 10-30 mM; non-limiting examples of the stabilizer are composed of sucrose at concentrations of about 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, 150 mM, 155 mM, 160 mM or 170 mM and L-arginine hydrochloride at concentrations of about 10 mM, 15 mM, 20 mM, 25 mM, or 30 mM.

[0231] In some embodiments, the stabilizer is L-arginine. In some embodiments, the stabilizer is L-arginine at a concentration of about 10-100 mM, preferably about 15-50 mM, more preferably about 20-30 mM. Non-limiting examples of L-arginine concentrations are about 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, preferably 25 mM or 30 mM.

[0232] In some embodiments, the stabilizer is methionine. In some embodiments, the stabilizer is methionine at a concentration of about 20-200 mM, preferably about 20-180 mM, preferably about 40-150 mM, preferably about 45-100 mM, preferably about 50-90 mM. Non-limiting examples of methionine concentrations are about 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 100 mM, preferably 60 mM or 65 mM.

[0233] In some embodiments, the stabilizer is sodium chloride. In some embodiments, the stabilizer is sodium chloride with a concentration of about 30-200 mM, preferably about 50-190 mM, preferably about 60-180 mM, preferably about 70-150 mM, preferably about 80-120 mM, and non-limiting examples of sodium chloride concentrations are about 60 mM, 70 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, preferably 100 mM or 105 mM.

[0234] In some embodiments, the stabilizer is L-arginine hydrochloride. In some embodiments, the stabilizer is L-arginine hydrochloride at a concentration of about 30-200 mM, preferably about 50-190 mM, more preferably about 100-180 mM, more preferably about 120-170 mM, more preferably about 130-150 mM. Non-limiting examples of L-arginine hydrochloride concentrations are about 100 mM, 110 mM, 120 mM, 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, 150 mM, 155 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, preferably 135 mM or 140 mM.

[0235] In some embodiments, the pharmaceutical composition further includes a surfactant selected from one or more of polysorbate 80, polysorbate 20, and poloxamer 188.

[0236] In some formulations, the surfactant is selected from polysorbate 80.

[0237] In some formulations, the surfactant mentioned above is selected from polysorbate 20.

[0238] In some embodiments, the concentration of the surfactant, calculated on a w / v basis, is approximately 0.001%-0.1%, preferably approximately 0.01%-0.1%, and more preferably approximately 0.01%-0.05%; as a non-limiting example, the concentration of the surfactant is approximately 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.09%, or 0.08%.

[0239] In some formulations, the above-mentioned pharmaceutical composition also includes water for injection.

[0240] In some formulations, the above-described pharmaceutical composition contains: (a) about 20 mg / mL to 280 mg / mL of any of the humanized anti-BLyS antibodies described herein or their antigen-binding fragments; (b) about 5-50 mM of histidine buffer or about 5-50 mM of acetate buffer, at a pH of about 5.0-6.5; (c) about 10-30 mM of L-arginine or about 10-30 mM of L-arginine hydrochloride; (d) about 80-220 mM of sucrose; and (e) about 0.01%-0.1% of polysorbate 80.

[0241] In some embodiments, the above-described pharmaceutical composition contains: (a) about 20 mg / mL to 280 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any of the embodiments herein; (b) about 10-40 mM of histidine buffer or about 10-40 mM of acetate buffer, at a pH of about 5.0-6.5; (c) about 10-30 mM of L-arginine or about 10-30 mM of L-arginine hydrochloride; (d) about 130-170 mM of sucrose; and (e) about 0.01%-0.1% of polysorbate 80.

[0242] In some embodiments, the above-described pharmaceutical composition contains: (a) about 30 mg / mL to 150 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any of the embodiments herein; (b) about 5-50 mM of histidine buffer or about 5-50 mM of acetate buffer, at a pH of about 5.0-6.5; (c) about 10-30 mM of L-arginine, about 10-30 mM of L-arginine hydrochloride or about 30-90 mM of methionine; (d) about 50-200 mM of sodium chloride; and (e) about 0.01%-0.1% of polysorbate 80.

[0243] In some embodiments, the pharmaceutical composition comprises: (a) about 30 mg / mL to 150 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any embodiment herein; (b) about 5-50 mM of histidine buffer, pH about 5.0-6.5; (c) about 10-30 mM of L-arginine, about 10-30 mM of L-arginine hydrochloride or about 30-90 mM of methionine; (d) about 50-200 mM of sodium chloride; and (e) about 0.01%-0.1% of polysorbate 80.

[0244] In some embodiments, the pharmaceutical composition comprises: (a) about 30 mg / mL to 150 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any embodiment herein; (b) about 5-50 mM of acetate buffer, pH about 5.0-6.5; (c) about 10-30 mM of L-arginine, about 10-30 mM of L-arginine hydrochloride or about 30-90 mM of methionine; (d) about 50-200 mM of sodium chloride; and (e) about 0.01%-0.1% of polysorbate 80.

[0245] In some embodiments, the above-described pharmaceutical composition contains: (a) about 80 mg / mL to 120 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any of the embodiments herein; (b) about 20-40 mM of histidine buffer or about 20-40 mM of acetate buffer, at a pH of about 5.5-6.0; (c) about 10-30 mM of L-arginine, about 50-70 mM of methionine, or about 10-30 mM of L-arginine hydrochloride; (d) about 80-120 mM of sodium chloride; and (e) about 0.01%-0.05% of polysorbate 80.

[0246] In some embodiments, the above-described pharmaceutical composition contains: (a) about 80 mg / mL to 120 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any of the embodiments herein; (b) about 20-40 mM of histidine buffer or at a pH of about 5.5-6.0; (c) about 10-30 mM of L-arginine, about 50-70 mM of methionine, or about 10-30 mM of L-arginine hydrochloride; (d) about 80-120 mM of sodium chloride; and (e) about 0.01%-0.05% of polysorbate 80.

[0247] In some embodiments, the above-described pharmaceutical composition contains: (a) about 80 mg / mL to 120 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any of the embodiments herein; (b) about 20-40 mM of acetate buffer, pH about 5.5-6.0; (c) about 10-30 mM of L-arginine, about 50-70 mM of methionine, or about 10-30 mM of L-arginine hydrochloride; (d) about 80-120 mM of sodium chloride; and (e) about 0.01%-0.05% of polysorbate 80.

[0248] In some formulations, the above-described pharmaceutical composition comprises: (a) about 180 mg / mL to 220 mg / mL of any of the humanized anti-BLyS antibodies described herein or their antigen-binding fragments; (b) about 20-40 mM of histidine buffer or about 20-40 mM of acetate buffer, pH about 5.5-6.0; (c) about 10-30 mM of L-arginine or about 10-30 mM of L-arginine hydrochloride; (d) about 130-170 mM of sucrose; and (e) about 0.01%-0.05% of polysorbate 80; or

[0249] In some embodiments, the above-described pharmaceutical composition contains: (a) about 200 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any embodiment herein; (b) about 25 mM histidine buffer, about 25 mM acetate buffer, or about 35-36 mM histidine buffer, at a pH of about 5.5-6.0; (c) about 15 mM L-arginine or about 15 mM L-arginine hydrochloride; (d) 150 mM sucrose; and (e) about 0.02% polysorbate 80.

[0250] In some embodiments, the above-described pharmaceutical composition contains: (a) about 100 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any embodiment herein; (b) about 25 mM histidine buffer, pH about 5.5-6.0; (c) about 25 mM L-arginine or about 25 mM L-arginine hydrochloride; (d) about 100 mM sodium chloride; and (e) about 0.02% polysorbate 80.

[0251] In some embodiments, the above-described pharmaceutical composition contains: (a) about 100 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any embodiment herein; (b) about 25 mM acetate buffer, pH about 5.5-6.0; (c) about 60 mM methionine; (d) about 100 mM sodium chloride; and (e) about 0.02% polysorbate 80.

[0252] In some embodiments, the above-described pharmaceutical composition contains: (a) about 100 mg / mL of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any embodiment herein; (b) about 35-36 mM histidine buffer at a pH of about 5.5-6.0; (c) about 15 mM L-arginine hydrochloride or about 15 mM L-arginine; (d) about 100 mM sodium chloride; and (e) about 0.02% polysorbate 80.

[0253] In some embodiments, the above pharmaceutical composition comprises any one of the components shown in (1)-(20) below:

[0254] (1)(a) approximately 20 mg / mL to 280 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; (b) approximately 5-50 mM histidine buffer, pH approximately 5.0-6.5; (c) approximately 10-30 mM L-arginine or approximately 10-30 mM L-arginine hydrochloride; (d) approximately 80-220 mM sucrose; (e) and approximately 0.01%-0.1% polysorbate 80; or

[0255] (2)(a) approximately 30 mg / mL to 150 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; (b) approximately 5-50 mM histidine buffer, pH approximately 5.0-6.5; (c) approximately 10-30 mM L-arginine; (d) approximately 50-200 mM sodium chloride; (e) and approximately 0.01%-0.1% polysorbate 80; or

[0256] (3)(a) approximately 30 mg / mL to 150 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; (b) approximately 5-50 mM histidine buffer, pH approximately 5.0-6.5; (c) approximately 10-30 mM L-arginine hydrochloride; (d) approximately 50-200 mM sodium chloride; (e) and approximately 0.01%-0.1% polysorbate 80; or

[0257] (4)(a) approximately 30 mg / mL to 150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; (b) approximately 5-50 mM histidine buffer, pH approximately 5.0-6.5; (c) approximately 30-90 mM methionine; (d) approximately 50-200 mM sodium chloride; (e) and approximately 0.01%-0.1% polysorbate 80; or

[0258] (5)(a) approximately 30 mg / mL to 150 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; (b) approximately 5-50 mM acetate buffer, pH approximately 5.0-6.5; (c) approximately 10-30 mM of L-arginine; (d) approximately 50-200 mM of sodium chloride; (e) and approximately 0.01%-0.1% of polysorbate 80; or

[0259] (6)(a) approximately 30 mg / mL to 150 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; (b) approximately 5-50 mM acetate buffer, pH approximately 5.0-6.5; (c) approximately 10-30 mM L-arginine hydrochloride; (d) approximately 50-200 mM sodium chloride; (e) and approximately 0.01%-0.1% polysorbate 80; or

[0260] (7)(a) approximately 30 mg / mL to 150 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 15; (b) approximately 5-50 mM acetate buffer, pH approximately 5.0-6.5; (c) approximately 30-90 mM methionine; (d) approximately 50-200 mM sodium chloride; (e) and approximately 0.01%-0.1% polysorbate 80; or

[0261] (8)(a) approximately 80 mg / mL to 120 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) approximately 20-40 mM histidine buffer, pH approximately 5.5-6.0; (c) approximately 10-30 mM L-arginine; (d) approximately 80-120 mM sodium chloride; (e) and approximately 0.01%-0.05% polysorbate 80; or

[0262] (9)(a) approximately 80 mg / mL to 120 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) approximately 20-40 mM histidine buffer, pH approximately 5.5-6.0; (c) approximately 10-30 mM L-arginine hydrochloride; (d) approximately 80-120 mM sodium chloride; (e) and approximately 0.01%-0.05% polysorbate 80; or

[0263] (10) (a) approximately 80 mg / mL to 120 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) approximately 20-40 mM acetate buffer, pH approximately 5.5-6.0; (c) approximately 50-70 mM methionine; (d) approximately 80-120 mM sodium chloride; (e) and approximately 0.01%-0.05% polysorbate 80; or

[0264] (11)(a) approximately 180 mg / mL to 220 mg / mL of a humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) approximately 20-40 mM histidine buffer, pH approximately 5.5-6.0; (c) approximately 10-30 mM of L-arginine or approximately 10-30 mM of L-arginine hydrochloride; (d) approximately 130-170 mM of sucrose; (e) and approximately 0.01%-0.05% of polysorbate 80; or

[0265] (12)(a) about 200 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 35-36 mM histidine buffer, pH about 5.5-6.0; (c) about 15 mM L-arginine hydrochloride; (d) about 150 mM sucrose; (e) and about 0.02% polysorbate 80; or

[0266] (13)(a) about 200 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 35-36 mM histidine buffer, pH about 5.5-6.0; (c) about 15 mM L-arginine; (d) about 150 mM sucrose; (e) and about 0.02% polysorbate 80; or

[0267] (14)(a) about 200 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 25 mM histidine buffer, pH about 5.5-6.0; (c) about 15 mM L-arginine hydrochloride; (d) about 150 mM sucrose; (e) and about 0.02% polysorbate 80; or

[0268] (15)(a) about 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 25 mM histidine buffer, pH about 5.5-6.0; (c) about 25 mM L-arginine; (d) about 100 mM sodium chloride; (e) and about 0.02% polysorbate 80; or

[0269] (16)(a) about 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 25 mM histidine buffer, pH about 5.5-6.0; (c) about 25 mM L-arginine hydrochloride; (d) about 100 mM sodium chloride; (e) and about 0.02% polysorbate 80; or

[0270] (17)(a) about 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 25 mM acetate buffer, pH about 5.5-6.0; (c) about 60 mM methionine; (d) about 100 mM sodium chloride; (e) and about 0.02% polysorbate 80; or

[0271] (18)(a) about 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 35-36 mM histidine buffer, pH about 5.5-6.0; (c) about 15 mM L-arginine hydrochloride; (d) about 100 mM sodium chloride; (e) and about 0.02% polysorbate 80; or

[0272] (19)(a) about 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 35-36 mM histidine buffer, pH about 5.5-6.0; (c) about 15 mM L-arginine; (d) about 100 mM sodium chloride; (e) and about 0.02% polysorbate 80; or

[0273] (20)(a) about 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) about 35 mM histidine buffer, pH about 5.8; (c) about 15 mM L-arginine hydrochloride; (d) about 100 mM sodium chloride; and (e) about 0.02% polysorbate 80;

[0274] Preferably, in the above-mentioned pharmaceutical compositions, the light chain amino acid sequence of the humanized anti-BLyS antibody is shown in SEQ ID NO:18, and the heavy chain amino acid sequence is shown in SEQ ID NO:19.

[0275] In some embodiments, the pharmaceutical composition is a liquid formulation or a lyophilized formulation.

[0276] In some embodiments, the pharmaceutical composition is a liquid formulation.

[0277] The above-mentioned liquid or lyophilized formulations are stable at 2-8°C for at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months.

[0278] The above aqueous solution or lyophilized preparation is stable at 40°C for at least 7 days, at least 14 days, or at least 28 days.

[0279] Medical uses and methods

[0280] This invention provides the use of humanized anti-BLyS antibodies or antigen-binding fragments thereof as described in any of the embodiments herein, polynucleotides described herein, expression vectors described herein, host cells described herein, pharmaceutical compositions described herein, or injections described herein in the preparation of medicaments for the prevention and / or treatment of diseases caused by excessive B cell proliferation.

[0281] This invention provides humanized anti-BLyS antibodies or antigen-binding fragments thereof as described in any of the embodiments herein, polynucleotides described herein, expression vectors described herein, host cells described herein, pharmaceutical compositions described herein, or injections described herein for the prevention and / or treatment of diseases caused by excessive B cell proliferation.

[0282] The present invention provides a method for preventing and / or treating diseases caused by excessive B cell proliferation, comprising administering to a subject in need a humanized anti-BLyS antibody or its antigen-binding fragment as described in any of the embodiments herein, a polynucleotide as described herein, an expression vector as described herein, a host cell as described herein, a pharmaceutical composition as described herein, or an injection as described herein.

[0283] In this invention, diseases caused by excessive B cell proliferation include systemic lupus erythematosus, rheumatoid arthritis, ankylosing spondylitis, or B-cell lymphoma.

[0284] The present invention will be described below by way of specific embodiments. It should be understood that these embodiments are merely illustrative and are not intended to limit the scope of the invention. The methods and materials used in the embodiments are conventional methods and materials in the art, unless otherwise stated.

[0285] Detection methods

[0286] The main pathways of protein degradation are the formation of aggregates, lysis products, and charged variants. Size exclusion chromatography (SEC-HPLC) was used to determine the percentage of native forms (protein monomers) and aggregates, while cation exchange chromatography (CEX-HPLC) was used to determine the percentage of acidic and basic antibody forms.

[0287] Stability was assessed using the following parameters: (1) visual appearance and visible foreign matter; (2) protein content determination by ultraviolet spectrophotometry; (3) antibody monomer, polymer, or fragment content determination by SEC-HPLC; (4) antibody major charge, acid charge, or basic charge content determination by CEX-HPLC; (5) antibody molecular weight determination by NR-CE-SDS method; (6) antibody molecular weight determination by R-CE-SDS method; and (7) antibody binding activity determination by ELISA method.

[0288] (1) Protein content

[0289] Protein concentration was determined using a Thermo Biomate 3S UV spectrophotometer. The percentage extinction coefficient (E1%) was set to 1.47 (g / mL) - 1 cm⁻¹. Using the Biomate 3S instrument, the cuvettes were rinsed three times with ultrapure water, and then 150 μL of ultrapure water was added before measurement. Ultrapure water was used for blank calibration. Each sample was measured in duplicate, with each solution measured three times. Before each measurement, the cuvette was rinsed twice with 150 μL of the solution, and then 150 μL of the solution was used for the measurement. The concentration of the corresponding sample was calculated using the extinction coefficient and OD value.

[0290] (2) SEC-HPLC purity

[0291] SEC purity was determined using HPLC (Ultimate 3000, Dionex instrument) with an SEC column (TOSOH G4000sxl SEC 7.8mm ID x 30cm, 5μm). The mobile phase composition was 50mM PB / 300mM NaCl, pH 7.0±0.2. The relative percentages of the main peak, polymers, and oligomers were calculated using the area normalization method. See Table 1 below for details.

[0292] Table 1: SEC-HPLC Purity Determination Methods

[0293]

[0294]

[0295] (3) CEX-HPLC purity

[0296] Peak area normalization was used for quantitative analysis of the results, and the peak area percentages of K0, K1, and K2 were calculated (K0 is the peak with the highest peak area percentage, K1 is the first peak after K0, and the retention time difference between K2 and K1 is approximately 2.5 min). The sum of the peak area percentages of K0, K1, and K2 represents the lysine variant content (i.e., sample purity). The sum of the peak area percentages of all peaks before K0 represents the acidic peak content, and the sum of the peak area percentages of all peaks remaining after K0 (excluding K1 and K2) represents the basic peak content. See Table 2 below for details.

[0297] Table 2: CEX-HPLC Purity Determination Methods

[0298]

[0299] (4) Insoluble particles

[0300] Turn on the instrument and adjust the illuminance knob to set the illuminance to 1200 lx.

[0301] Take 20 test samples, remove the container label, wipe the outer wall of the container to make the container clean and transparent, and let it stand at room temperature for 12 hours.

[0302] Place the test sample at the edge of the light shield, gently rotate and invert the container to suspend any visible foreign matter that may be present in the solution (do not create bubbles in the solution), and observe it at a distance of about 25 cm against a black and white background for 20 seconds.

[0303] Example 1: Antibody Preparation

[0304] Antibody sequence design:

[0305] Starting with antibody H48L47 (light chain as shown in SEQ ID NO: 16 and heavy chain as shown in SEQ ID NO: 17), to reduce the immunogenicity of the antibody and improve its stability, the frame region (FR) of the antibody was mutated, and the following amino acid sequences were screened out: the FR region of the light chain L47 (SEQ ID NO: 16) was mutated to generate three sequences: L472 (SEQ ID NO: 18), L473 (SEQ ID NO: 20) and L474 (SEQ ID NO: 22). The FR region of the heavy chain H48 (SEQ ID NO: 17) was mutated to generate four sequences: H482 (SEQ ID NO: 19), H483 (SEQ ID NO: 21), H484 (SEQ ID NO: 23) and H485 (SEQ ID NO: 24).

[0306] The light and heavy chains were paired and combined to construct expression vectors. The antibody expression light and heavy chain combinations are shown in Table 3 below. Transient expression was performed using 293F cells. The protein was obtained after one-step affinity purification in a 100ml system. The resulting twenty humanized antibodies are shown in Table 4 below.

[0307] Table 3: Antibody Expression Light and Heavy Chain Combinations

[0308] H48 H482 H483 H484 H485 L47 H48L47 H482L47 H483L47 H484L47 H485L47 L472 H48L472 H482L472 H483L472 H484L472 H485L472 L473 H48L473 H482L473 H483L473 H484L473 H485L473 L474 H48L474 H482L474 H483L474 H484L474 H485L474

[0309] Table 4: Amino acid sequence of the variable region and full-length sequence of the antibody

[0310]

[0311]

[0312] Example 2: Binding ELISA method for determining the binding ability of antibody to human BLyS protein

[0313] Human BLyS was coated onto a hydrophobic ELISA plate, and the binding ability of the antibody to human BLyS protein was measured using the Binding ELISA method. The EC50 was calculated from the fitted curve. The test results are shown in Table 5 below.

[0314] Table 5: Binding ability of antibodies to human BlyS protein

[0315]

[0316]

[0317] The results showed that all 20 antibodies of the present invention bound to the recombinant human BLyS protein. The EC50 of the antibody modified by the frame region (FR) sequence was smaller or equivalent to that of H48L47.

[0318] Example 3: Determination of the ability of antibody to competitively inhibit the binding of human BLyS to human BR3 by Blocking ELISA

[0319] Human BLyS was coated onto a hydrophobic ELISA plate. The ability of antibodies modified with the frame region (FR) sequence to competitively inhibit the binding of human BLyS to human BR3 was measured using the Blocking ELISA method, and the IC50 was calculated from the fitted curve. The Binding ELISA results showed that the EC50 of the antibodies modified with the frame region (FR) sequence was smaller than that of the unmodified antibodies. Therefore, only combinations with modified light and heavy chains, namely H482L472, H483L473, and H484L474, were selected for Blocking ELISA and compared with the commercially available positive control antibody Benlysta. The test results are shown in Table 6 below.

[0320] Positive control: Benlysta, Human Genome Sciences, patents see CN1279055C, CN106661111A.

[0321] Table 6: Ability of antibodies to competitively inhibit the binding of human BlyS to human BR3

[0322] Antibody name IC50 (μg / mL) H482L472 1.238 H483L473 0.8576 H484L474 1.275 Benlysta 4.714

[0323] The test results show that both the antibody modified with the frame region (FR) sequence and the positive control drug Benlysta can inhibit the binding of BLyS to its receptor BR3. The IC50 of antibodies H482L472, H483L473 and H484L474 are significantly better than that of the control Benlysta.

[0324] Example 4: Reporter gene assay to determine downstream NF-κB signaling stimulated by antibody competitive inhibition of human BLyS binding to TACI.

[0325] Human soluble BlyS binds to TACI, stimulating the downstream NF-κB signaling pathway. The downstream luciferase then releases a signal that is detected. Blocking the binding of BlyS and TACI with an antibody inhibits the downstream NF-κB signal. Changes in the signal value can be detected using a luciferase assay kit, and the IC50 is calculated from the fitted curve. Similarly, combinations with modified light and heavy chains, namely H482L472, H483L473, and H484L474, were selected for cell biology assays and compared with the commercially available positive control antibody Benlysta. The test results are shown in Table 7 below.

[0326] Positive control: Benlysta, Human Genome Sciences, patents see CN1279055C, CN106661111A.

[0327] Table 7: Antibody competitive inhibition of downstream NF-κB signaling stimulated by human BLyS binding to TACI

[0328] Antibody name IC 50 (ng / mL) H482L472 37.56 H483L473 29.27 H484L474 31.52 Benlysta 51.68

[0329] The test results show that both the modified frame region (FR) sequence antibody and the positive control drug Benlysta can inhibit the downstream NF-κB signaling stimulated by human BLyS binding to TACI. The IC50 values ​​of antibodies H482L472, H483L473 and H484L474 are significantly better than those of the control Benlysta.

[0330] Example 5: Buffer system and pH screening experiment

[0331] In liquid pharmaceutical compositions, the buffer system and pH closely affect antibody stability. Each antibody with unique physicochemical properties has an optimal type and pH of buffer solution. This embodiment aims to screen for an optimal buffer system and pH to ensure that the antibody disclosed in this invention has the best stability for clinical application. The antibody in this embodiment is H482L472.

[0332] 1.1 Buffer Preparation

[0333] Prepare 50 mL of each of the following solutions: 25 mM sodium acetate (pH 5.0, 5.5, 6.0), 25 mM sodium phosphate (pH 6.0, 6.5, 7.0), 35 mM histidine (pH 5.5, 6.0, 6.5), and 25 mM succinate buffer (pH 5.5, 6.0, 6.5).

[0334] 1.2 Fluid Change

[0335] Centrifugation and ultrafiltration were performed using 15 mL ultrafiltration tubes (Millipore, UCF803096) (5000 g × 15 min). 1 mL of protein was used per sample, and the medium was changed three times with 2 mL of buffer. After each medium change, the concentration was measured using a spectrophotometer, and the final concentration of the formulation was adjusted to approximately 30 mg / mL antibody.

[0336] 1.3 Sterilization filtration and sample placement

[0337] In a clean bench, the samples after liquid replacement were sterilized and filtered into 1.5 mL cryovials (approximately 1 mL per vial) using a syringe filter. The samples were incubated at 37°C for 7 days, and then analyzed by SEC and CEX-HPLC.

[0338] The experimental protocol is shown in Table 8 below.

[0339] Table 8: Buffer System and pH Screening Scheme

[0340]

[0341] The test results are shown in Table 9 below.

[0342] Table 9: Results of buffer system and pH screening tests

[0343]

[0344] The experimental results show that after being placed at 37℃ for 6 days, the SEC and CEX purities of acetate buffer and histidine buffer decreased less, which was better than that of phosphate and succinate buffers. Therefore, acetate and histidine buffers were selected for the next round of screening.

[0345] Example 6: Stabilizer Screening Experiment

[0346] According to literature reports, high protein concentrations in monoclonal antibody formulations often lead to adverse effects such as protein aggregation and increased polymer content. This experiment, based on the formulation buffer system selected in the first round of screening, added several different amino acids (L-arginine and methionine) to investigate the effects of these excipients on formulation stability.

[0347] 2.1 Buffer Preparation

[0348] The buffer solutions were prepared according to the formulations in Tables 10 and 11, respectively. The histidine buffer concentration was 35 mM; the acetate buffer concentration was 25 mM; the antibody used in this example was H482L472; the final antibody concentration in each formulation was 100 mg / mL; and the sodium chloride concentration in each formulation was 100 mM.

[0349] Table 10: Experimental protocol for stabilizer screening in histidine buffer system

[0350]

[0351] Table 11: Experimental scheme for stabilizer screening in acetate buffer system

[0352]

[0353]

[0354] 2.2 Fluid Change

[0355] Ultrafiltration concentration and buffer exchange were performed using a 50 mL centrifuge ultrafiltration tube (Millipore, UFC903096) (5000 g × 25 min). 3.5 mL of each sample was ultrafiltered and concentrated to 1 mL, then 4 mL of the corresponding buffer was added and the solution was exchanged three times. After the buffer exchange was completed, the volume was adjusted to 1 mL for later use. The final antibody concentration in the sample was approximately 100 mg / mL.

[0356] 2.3 Sterilization Filtration

[0357] After sterile filtration of the sample following the solution change in the laminar flow hood, it is dispensed into 1mL pre-filled syringes, approximately 1mL per syringe.

[0358] 2.4 Slotting and Testing

[0359] The sterilized filtered samples were incubated in a 37°C incubator for 7 days. SEC and CEX-HPLC were then performed.

[0360] The experimental results are shown in Tables 12 and 13 below.

[0361] Table 12: Results of stabilizer screening experiments in histidine buffer system

[0362]

[0363] Table 13: Results of stabilizer screening experiments in the acetate buffer system

[0364]

[0365]

[0366] Based on the experimental results, the pH of the formulation significantly affects its stability, regardless of whether it contains histidine or acetic acid. As the pH increases, the purity initially increases and then decreases. Adding L-arginine or methionine to the formulation effectively reduces aggregate formation. Based on these results, two formulations were selected: 35mM histidine + 15mM L-arginine + 100mM sodium chloride, pH 5.8; and 25mM acetic acid + 60mM methionine + 100mM sodium chloride, pH 5.8.

[0367] Example 7: Forced Degradation Experiment

[0368] Further forced degradation experiments were conducted on the selected formulations to examine their stability under various screening pressures. The antibody used in this example was H482L472, and the experimental protocol is shown in Table 14 below.

[0369] Table 14: Forced Degradation Experimental Protocol

[0370]

[0371] 3.1 Buffer Preparation

[0372] Prepare 500 mL of each of the required reagent buffer solutions according to the formulas in Table 15 below, filter them through a 0.2 μm filter and set aside.

[0373] Table 15: Formulation Buffer Formulation

[0374]

[0375] 3.2 Concentration and Liquid Replacement

[0376] Ultrafiltration concentration and buffer replacement were performed using 50 mL ultrafiltration tubes (Millipore, UFC903096) (5000 g × 25 min). 4.5 mL of the original solution was added to each tube, and after ultrafiltration concentration to approximately 1.5 mL, 4 mL of the corresponding buffer solution was added and the solution was replaced three times. After each buffer replacement, the mixture was thoroughly mixed, and the protein concentration was measured. The final sample concentration was adjusted to approximately 100 mg / mL.

[0377] 3.3 Preparation of Tween-80 stock solution

[0378] Prepare 10 mL of an aqueous solution containing 2% Tween-80 and shake to mix. Add an appropriate volume of the prepared Tween-80 stock solution to the sample after the solution has been changed, adjusting the Tween-80 content in the sample to 0.02%.

[0379] 3.4 Sterilization filtration and dispensing

[0380] After filtering the prepared sample through a 0.2μm sterile filter in the laminar flow hood, it is dispensed into 1mL pre-filled syringes, approximately 1mL per syringe.

[0381] 3.5 Layout and Testing

[0382] Slot the samples according to the plan in Table 14 and test them.

[0383] The experimental results are shown in Table 16-20 below.

[0384] Table 16: Results of the shaking experiment

[0385]

[0386] Table 17: Freeze-thaw test results

[0387]

[0388]

[0389] Table 18: Results of Ultraviolet Irradiation Experiment

[0390]

[0391] Table 19: Results of High-Temperature Accelerated Test for SEC / CEX Purity

[0392]

[0393] Table 20: Results of High-Temperature Accelerated Test for Appearance and Visible Foreign Matter

[0394]

[0395]

[0396] The experimental results show that no significant decrease in purity was observed after shaking and freeze-thaw cycles for formulations F1 and F2. However, after 8 hours of UV irradiation, the SEC and CEX purities of both formulations showed a decreasing trend, indicating that the formulations are sensitive to UV light and need to be stored in the dark. The high-temperature accelerated test showed that the appearance of the samples did not change significantly after being placed at high temperatures for 4W and 8W, but the acid peak and polymer content increased significantly. Among them, the increasing trend of formulation F1 was slightly lower than that of F2. During subsequent production according to Formulation F1, the pH needed to be adjusted with hydrochloric acid due to the slightly high alkalinity of L-arginine. Therefore, L-arginine in the original Formulation F1 was replaced with L-arginine hydrochloride (CAS No.: 1119-34-2). The stability of the replaced Formulation F3 (formulation: 100 mg / mL antibody + 35 mM histidine buffer + 15 mM L-arginine hydrochloride + 100 mM sodium chloride + 0.02% Tween-80, pH 5.8) was investigated. After being placed at 4℃ and 37℃ for 4 weeks, the purity of SEC and CEX was measured. The test results are shown in Table 21 below.

[0397] Table 21: Stability test results after replacing arginine hydrochloride

[0398]

[0399] The experimental results show that when arginine is replaced with arginine hydrochloride, the decreasing trend of SEC and CEX is basically the same as when arginine is used. The addition of the above two stabilizers to the pharmaceutical composition of the present invention can maintain the good stability of the prescription formulation.

[0400] Example 8: Compatibility Stability Study Experiment

[0401] 4.1 Preparation of physiological saline

[0402] Prepare 500 mL of 0.9% physiological saline using ultrapure water, and stir with a magnetic stirrer until completely dissolved.

[0403] 4.2 Sample Dilution

[0404] The formulation sample (formulation: 100 mg / mL antibody + 35 mM histidine buffer + 15 mM L-arginine hydrochloride + 100 mM sodium chloride + 0.02% Tween-80, pH 5.8) was diluted to the corresponding final concentration (60, 30, 10, 3 mg / mL) using the physiological saline prepared in 5.4.1.1. The volume of the diluted sample was approximately 25 mL. The sample was thoroughly shaken before use. The antibody used in this example was H482L472.

[0405] 4.3 Sample Packaging

[0406] After being sterilized and filtered in a laminar flow hood, the diluted samples were aseptically dispensed into 6 mL vials. Each concentration of sample was dispensed into 5 vials, each containing 5 mL. The vials were then capped using a crimping machine.

[0407] 4.4 Layout and Testing

[0408] Samples from each group were incubated at 25℃ and 30℃ for 0, 2, and 4 hours, respectively, and then stored at -80℃. After being frozen at -80℃ for 5 days, the samples were sent for testing to determine SEC purity, biological activity (blocking ELISA), and insoluble microparticles.

[0409] The experimental results are shown in Tables 22 and 23 below.

[0410] Table 22: Detection results after incubation at 25℃

[0411]

[0412]

[0413] Table 23: Detection results after incubation at 30℃

[0414]

[0415] The experimental results show that after incubation at 25℃ and 30℃ for 2 hours and 4 hours, respectively, the test results for each concentration group did not show a significant decrease, and all met the requirements of the product quality standards. Therefore, it can be concluded that the formulation is stable within 4 hours of storage at room temperature after dilution with physiological saline.

[0416] Compatibility stability tests showed that the final formulation, diluted with 0.9% sodium chloride solution, remained stable at room temperature for 4 hours within a protein concentration range of 3–60 mg / ml.

[0417] In summary, by investigating different buffer systems, pH conditions, antibody concentrations, and excipient compositions, the optimal formulation was finally determined. Histidine buffer was used to adjust the pH, arginine hydrochloride and sodium chloride were used to adjust the osmotic pressure of the formulation, and polysorbate 20 was added to increase the solubility of the formulation.

[0418] Example 9: Study on the inhibitory effect of UBP1213sc formulation on physiological changes induced by human BLyS in Balb / c mice

[0419] Animals were randomly divided into 7 groups of 10 each, based on their weight: G1 The animals were divided into four groups: G2 (Vehicle model control group), G3 (KLH hIgG1 group), G4 (UBP1213sc 5mg / kg group), G5 (UBP1213sc 1.5mg / kg group), G6 (UBP1213sc 0.5mg / kg group), and G7 (UBP1213sc 0.15mg / kg group). Administration was performed on Day 1 and Day 3, for a total of two administrations. From Day 1 to Day 4, 0.3mg / kg of human BlyS was administered subcutaneously. Animal weight was monitored daily during the experiment. At the end of the experiment, animals were euthanized, serum IgA levels were measured, spleens were harvested and weighed to calculate organ coefficients, and B220 levels in the spleen were measured using FACS. + / ThB + Cell proportions. The UBP1213sc formulation consisted of: 100 mg / mL antibody H482L472 + 35 mM histidine buffer + 15 mM L-arginine hydrochloride + 100 mM sodium chloride + 0.02% Tween-80, pH 5.8. Test results are shown in Tables 24-26.

[0420] Table 24: Effects of UBP1213sc on spleen wet weight and spleen organ coefficient in BLyS-stimulated mice (data expressed as mean ± SEM, n = 10)

[0421]

[0422] Note: * :P<0.05; ** :P<0.01; *** :P<0.01vs # :P<0.05; ## :P<0.01; ### :P<0.01vsVehicle.

[0423] Table 25: B220 in the spleen of mice stimulated by UBP1213sc and BlyS + / ThB + The impact of proportions (data expressed as mean ± SEM, n = 10)

[0424]

[0425]

[0426] Note: * :P<0.05; ** :P<0.01; *** :P<0.01vs # :P<0.05; ## :P<0.01; ### :P<0.01vsVehicle.

[0427] Table 26: Effect of UBP1213sc on serum IgA levels in BLyS-stimulated mice (data expressed as mean ± SEM, n = 10)

[0428]

[0429] Note: * :P<0.05; **:P<0.01; ***:P<0.001vs # :P<0.05; ## :P<0.01; ### :P<0.01vs Vehicle.

[0430] According to the experimental results in Table 24-26, compared with Compared to the control group, the Vehicle group showed that 0.3 mg / kg human BLyS (subcutaneous injection) successfully induced a significant increase in spleen wet weight / spleen organ coefficient, spleen B220+ / ThB+ cell ratio, and serum IgA level in Balb / c mice. Compared to the Vehicle group, it significantly inhibited the increase in various physiological indicators in the model mice; 1.5 mg / kg UBP1213 subcutaneously significantly inhibited the increase in various physiological indicators in the model mice; 0.5, 1.5, and 5 mg / kg UBP1213sc significantly inhibited the increase in various physiological indicators in the model mice.

[0431] Subcutaneous injection of human BLyS induced a significant increase in spleen wet weight / spleen organ coefficient, spleen B220+ / ThB+ cell ratio, and serum IgA levels in Balb / c mice. Subcutaneous injection of different doses (0.15, 0.5, 1.5, 5 mg / kg) of the anti-BLyS monoclonal antibody UBP1213sc significantly inhibited the increase in spleen wet weight / spleen organ coefficient in mice. UBP1213sc at 1.5 and 5 mg / kg also significantly inhibited the increase in B220+ / ThB+ cell ratio and serum IgA levels, exhibiting a favorable dose-response relationship. Furthermore, there were no significant changes in body weight among the treated groups. sequence list <110> Shanghai Junshi Biosciences Co., Ltd. Suzhou Junmeng Biomedical Technology Co., Ltd. <120> Anti-BLyS antibodies, their pharmaceutical compositions and their uses <130> 213020Z1 1CNCN <150> CN 202110441100.8 <151> 2021-04-23 <160> twenty four <170> PatentIn version 3.5 <210> 1 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> LCDR1 <400> 1 Arg Thr Ser Gln Asp Ile Ser Ile Tyr Leu Asn 1 5 10 <210> 2 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> LCDR2 <400> 2 Tyr Thr Ser Arg Leu Arg Ser 1 5 <210> 3 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> LCDR3 <400> 3 Gln Gln Gly Ser Thr Leu Pro Trp Thr 1 5 <210> 4 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> HCDR1 <400> 4 Asp Ala Trp Met Asp 1 5 <210> 5 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> HCDR2 <400> 5 Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu Ser 1 5 10 15 Val Lys Gly <210> 6 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> HCDR3 <400> 6 Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr Ile Met Asp Tyr 1 5 10 15 <210> 7 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> VL <400> 7 Asp Ile Gln Met Thr Gln Thr Pro Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 8 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> VH <400> 8 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 125 <210> 9 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> VL <400> 9 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 10 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> VH <400> 10 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 11 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> VL <400> 11 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 12 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> VH <400> 12 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 13 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> VL <400> 13 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 14 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> VH <400> 14 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 15 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> VH <400> 15 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 16 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> LC <400> 16 Asp Ile Gln Met Thr Gln Thr Pro Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 17 <211> 457 <212> PRT <213> Artificial Sequence <220> <223> HC <400> 17 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala 115 120 125 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 130 135 140 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 145 150 155 160 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 165 170 175 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 180 185 190 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 195 200 205 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg 210 215 220 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 225 230 235 240 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 245 250 255 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 260 265 270 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 275 280 285 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 290 295 300 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 305 310 315 320 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 325 330 335 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 340 345 350 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 355 360 365 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 370 375 380 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 385 390 395 400 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 405 410 415 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 420 425 430 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 435 440 445 Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 <210> 18 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> LC <400> 18 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 19 <211> 457 <212> PRT <213> Artificial Sequence <220> <223> HC <400> 19 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 115 120 125 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 130 135 140 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 145 150 155 160 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 165 170 175 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 180 185 190 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 195 200 205 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg 210 215 220 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 225 230 235 240 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 245 250 255 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 260 265 270 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 275 280 285 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 290 295 300 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 305 310 315 320 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 325 330 335 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 340 345 350 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 355 360 365 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 370 375 380 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 385 390 395 400 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 405 410 415 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 420 425 430 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 435 440 445 Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 <210> 20 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> LC <400> 20 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 21 <211> 457 <212> PRT <213> Artificial Sequence <220> <223> HC <400> 21 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 115 120 125 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 130 135 140 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 145 150 155 160 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 165 170 175 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 180 185 190 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 195 200 205 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg 210 215 220 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 225 230 235 240 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 245 250 255 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 260 265 270 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 275 280 285 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 290 295 300 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 305 310 315 320 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 325 330 335 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 340 345 350 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 355 360 365 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 370 375 380 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 385 390 395 400 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 405 410 415 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 420 425 430 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 435 440 445 Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 <210> 22 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> LC <400> 22 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Asp Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Ser Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 23 <211> 457 <212> PRT <213> Artificial Sequence <220> <223> HC <400> 23 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 115 120 125 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 130 135 140 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 145 150 155 160 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 165 170 175 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 180 185 190 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 195 200 205 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg 210 215 220 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 225 230 235 240 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 245 250 255 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 260 265 270 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 275 280 285 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 290 295 300 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 305 310 315 320 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 325 330 335 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 340 345 350 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 355 360 365 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 370 375 380 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 385 390 395 400 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 405 410 415 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 420 425 430 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 435 440 445 Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455 <210> 24 <211> 457 <212> PRT <213> Artificial Sequence <220> <223> HC <400> 24 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala 20 25 30 Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Glu Ile Arg Asn Lys Ala Asn Asn His Ala Thr Tyr Gln Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Ala Pro Phe Asp Leu Leu Val Arg Arg Gly Tyr Tyr 100 105 110 Ile Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 115 120 125 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 130 135 140 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 145 150 155 160 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 165 170 175 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 180 185 190 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 195 200 205 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg 210 215 220 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 225 230 235 240 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 245 250 255 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 260 265 270 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 275 280 285 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 290 295 300 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 305 310 315 320 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 325 330 335 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 340 345 350 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 355 360 365 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 370 375 380 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 385 390 395 400 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 405 410 415 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 420 425 430 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 435 440 445 Lys Ser Leu Ser Leu Ser Pro Gly Lys 450 455

Claims

1. A humanized anti-BLyS antibody or its antigen-binding fragment, comprising a heavy chain variable region and a light chain variable region, characterized in that, The light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 9, and the heavy chain variable region contains an amino acid sequence as shown in SEQ ID NO:

10.

2. The humanized anti-BLyS antibody or its antigen-binding fragment as described in claim 1, wherein, The humanized anti-BLyS antibody further contains a human light chain constant region and a heavy chain constant region, and the light chain variable region and the heavy chain variable region are respectively connected to the human light chain constant region and the heavy chain constant region.

3. The humanized anti-BLyS antibody or its antigen-binding fragment as described in claim 2, characterized in that, The constant region of the human light chain is the constant region of the human light chain κ.

4. The humanized anti-BLyS antibody or its antigen-binding fragment as described in any one of claims 1-3, comprising a light chain and a heavy chain, wherein, The light chain comprises the amino acid sequence shown in SEQ ID NO: 18, and the heavy chain comprises the amino acid sequence shown in SEQ ID NO:

19.

5. A polynucleotide molecule selected from polynucleotide molecules encoding a humanized anti-BLyS antibody or an antigen-binding fragment thereof as described in any one of claims 1-4.

6. An expression vector comprising the polynucleotide molecule as described in claim 5.

7. The expression vector as described in claim 6, characterized in that, The expression vector is a eukaryotic expression vector.

8. A host cell comprising the polynucleotide molecule as described in claim 5 or the expression vector as described in claim 6 or 7.

9. The host cell as described in claim 8, characterized in that, The host cell is a eukaryotic cell.

10. The host cell as described in claim 9, characterized in that, The host cell is a mammalian cell.

11. A pharmaceutical composition comprising: (1) Buffer solution, wherein the buffer solution is selected from one or more of acetate buffer, succinate buffer, phosphate buffer and histidine buffer; (2) Humanized anti-BLyS antibody or its antigen-binding fragment, wherein, The humanized anti-BLyS antibody or its antigen-binding fragment is as described in any one of claims 1-4; (3) A stabilizer, wherein the stabilizer is selected from one or more of arginine, arginine hydrochloride, methionine, and sodium chloride; and (4) Surfactant, wherein the surfactant is polysorbate 80 or polysorbate 20.

12. The pharmaceutical composition of claim 11, wherein, The concentration of the humanized anti-BLyS antibody or its antigen-binding fragment is 10-300 mg / mL.

13. The pharmaceutical composition according to claim 12, characterized in that, The concentration of the humanized anti-BLyS antibody or its antigen-binding fragment is 20-280 mg / mL.

14. The pharmaceutical composition according to claim 12, characterized in that, The concentration of the humanized anti-BLyS antibody or its antigen-binding fragment is 30-150 mg / mL.

15. The pharmaceutical composition according to claim 12, characterized in that, The concentration of the humanized anti-BLyS antibody or its antigen-binding fragment is 80-120 mg / mL.

16. The pharmaceutical composition of claim 12, characterized in that, The concentration of the humanized anti-BLyS antibody or its antigen-binding fragment is 180-220 mg / mL.

17. The pharmaceutical composition of claim 11, characterized in that, The concentration of the buffer solution is 5-50 mM.

18. The pharmaceutical composition of claim 17, characterized in that, The concentration of the buffer solution is 10-40 mM.

19. The pharmaceutical composition of claim 17, characterized in that, The concentration of the buffer solution is 20-40 mM.

20. The pharmaceutical composition of claim 17, characterized in that, The pH of the buffer solution is 5.0-6.

5.

21. The pharmaceutical composition of claim 20, characterized in that, The pH of the buffer solution is 5.0-6.

0.

22. The pharmaceutical composition of claim 20, characterized in that, The pH of the buffer solution is 5.5-6.

0.

23. The pharmaceutical composition of claim 11, characterized in that, The concentration of the stabilizer is 50 mM to 300 mM.

24. The pharmaceutical composition according to claim 23, characterized in that, The concentration of the stabilizer is 100 mM to 300 mM.

25. The pharmaceutical composition according to claim 23, characterized in that, The concentration of the stabilizer is 100 mM to 200 mM.

26. The pharmaceutical composition of claim 11, characterized in that, The concentration of the surfactant is 0.001%-0.1% (w / v).

27. The pharmaceutical composition of claim 26, characterized in that, The concentration of the surfactant is 0.01%-0.1% (w / v).

28. The pharmaceutical composition of claim 26, characterized in that, The concentration of the surfactant is 0.01%-0.05% (w / v).

29. The pharmaceutical composition of claim 11, wherein, The stabilizer is a combination of 10-30 mM L-arginine hydrochloride and 50-200 mM sodium chloride; or the stabilizer is a combination of 10-30 mM L-arginine and 50-200 mM sodium chloride; or the stabilizer is a combination of 30-90 mM methionine and 50-200 mM sodium chloride; or the stabilizer is 10-120 mM L-arginine hydrochloride; or the stabilizer is 10-120 mM methionine.

30. The pharmaceutical composition of claim 29, characterized in that, The stabilizer is a combination of 10-30 mM L-arginine hydrochloride and 80-120 mM sodium chloride; or the stabilizer is a combination of 10-30 mM L-arginine and 80-120 mM sodium chloride; or the stabilizer is a combination of 50-70 mM methionine and 80-120 mM sodium chloride.

31. The pharmaceutical composition according to claim 11, characterized in that, The pharmaceutical composition comprises: (a) 30 mg / mL to 150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment; (b) 5-50 mM histidine buffer or 5-50 mM acetate buffer, pH 5.0-6.5; (c) 10-30 mM L-arginine, 10-30 mM L-arginine hydrochloride or 30-90 mM methionine; (d) 50-200 mM sodium chloride; and (e) 0.01%-0.1% polysorbate 80.

32. The pharmaceutical composition according to claim 31, characterized in that, The pharmaceutical composition comprises: (a) 80 mg / mL-120 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment; (b) 20-40 mM histidine buffer or 20-40 mM acetate buffer, pH 5.5-6.0; (c) 10-30 mM L-arginine, 10-30 mM L-arginine hydrochloride or 50-70 mM methionine; (d) 80-120 mM sodium chloride; and (e) 0.01%-0.05% polysorbate 80.

33. The pharmaceutical composition according to claim 31, characterized in that, The pharmaceutical composition comprises: (a) 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment; (b) 25 mM histidine buffer at pH 5.5-6.0; (c) 25 mM L-arginine or 25 mM L-arginine hydrochloride; (d) 100 mM sodium chloride; and (e) 0.02% polysorbate 80.

34. The pharmaceutical composition according to claim 31, characterized in that, The pharmaceutical composition contains: (a) 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment; (b) 25 mM acetate buffer, pH 5.5-6.0; (c) 60 mM methionine; (d) 100 mM sodium chloride; and (e) 0.02% polysorbate 80.

35. The pharmaceutical composition according to claim 31, characterized in that, The pharmaceutical composition comprises: (a) 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment; (b) 35-36 mM histidine buffer at pH 5.5-6.0; (c) 15 mM L-arginine hydrochloride or 15 mM L-arginine; (d) 100 mM sodium chloride; and (e) 0.02% polysorbate 80.

36. The pharmaceutical composition of claim 31, comprising any one of the components shown in (1)-(6) below: (1) (a) 30 mg / mL-150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) 5-50 mM histidine buffer, pH 5.0-6.5; (c) 10-30 mM L-arginine; (d) 50-200 mM sodium chloride; (e) and 0.01%-0.1% polysorbate 80; or (2) (a) 30 mg / mL-150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, wherein the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 9 and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 10; (b) 5-50 mM histidine buffer, pH 5.0-6.5; (c) 10-30 mM L-arginine hydrochloride; (d) 50-200 mM sodium chloride; (e) and 0.01%-0.1% polysorbate 80; or (3) (a) 30 mg / mL-150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, wherein the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 9 and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 10; (b) 5-50 mM histidine buffer, pH 5.0-6.5; (c) 30-90 mM methionine; (d) 50-200 mM sodium chloride; (e) and 0.01%-0.1% polysorbate 80; or (4) (a) 30 mg / mL-150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, wherein the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 9 and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 10; (b) 5-50 mM acetate buffer, pH 5.0-6.5; (c) 10-30 mM L-arginine; (d) 50-200 mM sodium chloride; (e) and 0.01%-0.1% polysorbate 80; or (5) (a) 30 mg / mL-150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, wherein the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 9 and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 10; (b) 5-50 mM acetate buffer, pH 5.0-6.5; (c) 10-30 mM L-arginine hydrochloride; (d) 50-200 mM sodium chloride; (e) and 0.01%-0.1% polysorbate 80; or (6) (a) 30 mg / mL to 150 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, wherein the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 9 and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 10; (b) 5-50 mM acetate buffer, pH 5.0-6.5; (c) 30-90 mM methionine; (d) 50-200 mM sodium chloride; and (e) 0.01%-0.1% polysorbate 80.

37. The pharmaceutical composition of claim 31, comprising any one of the components shown in (7)-(9) below: (7) (a) 80 mg / mL-120 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, wherein the humanized anti-BLyS antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 9 and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 10; (b) 20-40 mM histidine buffer, pH 5.5-6.0; (c) 10-30 mM L-arginine; (d) 80-120 mM sodium chloride; (e) and 0.01%-0.05% polysorbate 80; or (8) (a) 80 mg / mL-120 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) 20-40 mM histidine buffer, pH 5.5-6.0; (c) 10-30 mM L-arginine hydrochloride; (d) 80-120 mM sodium chloride; (e) and 0.01%-0.05% polysorbate 80; or (9) (a) 80 mg / mL-120 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment, the humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, and the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) 20-40 mM acetate buffer, pH 5.5-6.0; (c) 50-70 mM methionine; (d) 80-120 mM sodium chloride; and (e) 0.01%-0.05% polysorbate 80.

38. The pharmaceutical composition of claim 31, comprising any one of the components shown in (10)-(12): (10) (a) 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 9, said heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 10; (b) 25 mM histidine buffer, pH 5.5-6.0; (c) 25 mM L-arginine; (d) 100 mM sodium chloride; (e) and 0.02% polysorbate 80; or (11) (a) 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) 25 mM histidine buffer, pH 5.5-6.0; (c) 25 mM L-arginine hydrochloride; (d) 100 mM sodium chloride; (e) and 0.02% polysorbate 80; or (12) (a) 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, the humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, and the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) 25 mM acetate buffer, pH 5.5-6.0; (c) 60 mM methionine; (d) 100 mM sodium chloride; and (e) 0.02% polysorbate 80.

39. The pharmaceutical composition of claim 31, comprising the components shown in (13) or (14) below: (13) (a) 100 mg / mL of humanized anti-BLyS antibody or its antigen-binding fragment thereof, said humanized anti-BLyS antibody or its antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 9, said heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 10; (b) 35-36 mM histidine buffer, pH 5.5-6.0; (c) 15 mM L-arginine hydrochloride; (d) 100 mM sodium chloride; (e) and 0.02% polysorbate 80; or (14) (a) 100 mg / mL of humanized anti-BLyS antibody or antigen-binding fragment thereof, the humanized anti-BLyS antibody or antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, and the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) 35-36 mM histidine buffer, pH 5.5-6.0; (c) 15 mM L-arginine; (d) 100 mM sodium chloride; and (e) 0.02% polysorbate 80.

40. The pharmaceutical composition of claim 31, comprising the following components: (a) 100 mg / mL humanized anti-BLyS antibody or antigen-binding fragment thereof, said humanized anti-BLyS antibody or antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 10; (b) 35 mM histidine buffer at pH 5.8; (c) 15 mM L-arginine hydrochloride; (d) 100 mM sodium chloride; and (e) 0.02% polysorbate 80.

41. The pharmaceutical composition according to any one of claims 36-40, characterized in that, The light chain amino acid sequence of the humanized anti-BLyS antibody is shown in SEQ ID NO:18, and the heavy chain amino acid sequence is shown in SEQ ID NO:

19.

42. An injectable preparation comprising the pharmaceutical composition of any one of claims 11-41 and a sodium chloride solution.

43. The injectable formulation as described in claim 42, characterized in that, In the injectable preparation, the concentration of the humanized anti-BLyS antibody is 3~100 mg / mL.

44. The injectable formulation as described in claim 43, characterized in that, The pH of the injection is 5.5 to 6.

0.

45. The pharmaceutical composition according to any one of claims 11-41 or the injectable preparation according to any one of claims 42-44, administered by subcutaneous injection.

46. ​​The use of the humanized anti-BLyS antibody or its antigen-binding fragment as described in any one of claims 1-4, or the polynucleotide molecule as described in claim 5, or the expression vector as described in claim 6 or 7, or the host cell as described in any one of claims 8-10, or the pharmaceutical composition as described in any one of claims 11-41, or the injection as described in any one of claims 42-44 in the preparation of a medicament for the prevention and / or treatment of diseases caused by excessive B cell proliferation, wherein the disease caused by excessive B cell proliferation is systemic lupus erythematosus.

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