Pharmaceutical formulations comprising anti-alpha4beta7 antibodies

Abstract

The invention provides a stable pharmaceutical preparation which comprises 60 g / L of an anti-alpha4beta7 antibody, 112 g / L of trehalose, 26.34 g / L of arginine hydrochloride, 0.6 g / L of Tween 80 and 50 mM of a histidine-histidine hydrochloric acid buffer solution, the PH value of the pharmaceutical preparation is 6.3, and the anti-alpha4beta7 antibody is videlizumab.

Description

Technical Field

[0001] The present invention relates to the field of biological pharmaceutical preparations, and in particular, to a stable pharmaceutical preparation containing vedolizumab. Background Art

[0002] Vedolizumab, a humanized monoclonal antibody, specifically binds to α4β7 integrin, blocks the interaction between α4β7 integrin and MAdCAM-1, and inhibits the migration of memory T lymphocytes across the endothelium to the inflammatory tissue of the gastrointestinal tract, thereby accurately inhibiting intestinal inflammation. It is often used to treat ulcerative colitis and Crohn's disease.

[0003] Currently marketed pharmaceutical products It is a lyophilized Vidali beads preparation, which after reconstitution is 60 mg / ml antibody, 50 mM histidine, 10% (w / v) sucrose, 125 mM arginine hydrochloride and 0.06% (w / v) polysorbate 80, with a pH of 6.3. On this basis, the stability of the antibody in the preparation is further improved, and it is necessary to ensure that the dosage and product safety requirements are met when the molecule is administered to patients, and to minimize protein degradation, especially protein aggregation, to avoid severe immunogenic reactions as much as possible.

[0004] Therefore, there is still a need in the art to optimize the formulation of Vidli beads, improve antibody stability, and enhance storage capacity. Summary of the invention

[0005] The object of the present invention is to provide an optimized preparation comprising an anti-α4β7 antibody, which exhibits improved physical and chemical stability during long-term storage.

[0006] The first aspect of the present invention provides a stable pharmaceutical preparation comprising an anti-α4β7 antibody and a first stabilizer, wherein the first stabilizer is trehalose.

[0007] In some embodiments, the concentration of trehalose is about 99-118 g / L.

[0008] In some embodiments, the concentration of trehalose is about 99-112 g / L.

[0009] In some embodiments, the concentration of trehalose is about 112 g / L.

[0010] In some embodiments, the anti-α4β7 antibody comprises a heavy chain variable region (VH) and, the anti-α4β7 antibody comprises a light chain variable region (VL); wherein the heavy chain variable region comprises HCDR1 with an amino acid sequence of SEQ ID NO:1, HCDR2 with an amino acid sequence of SEQ ID NO:2 and / or HCDR3 with an amino acid sequence of SEQ ID NO:3; the light chain variable region comprises LCDR1 with an amino acid sequence of SEQ ID NO:4, LCDR2 with an amino acid sequence of SEQ ID NO:5 and / or LCDR3 with an amino acid sequence of SEQ ID NO:6.

[0011] SEQ ID NO: 1:

[0012] Ser Tyr Trp Met His

[0013] SEQ ID NO:2:

[0014] Glu Ile Asp Pro Ser Glu Ser Asn Thr Asn Tyr Asn Gln Lys Phe Lys

[0015] SEQ ID NO:3:

[0016] Gly Gly Tyr Asp Gly Trp Asp Tyr Ala Ile Asp Tyr

[0017] SEQ ID NO:4:

[0018] Arg Ser Ser Gln Ser Leu Ala Lys Ser Tyr Gly Asn Thr Tyr Leu Ser

[0019] SEQ ID NO:5

[0020] Gly Ile Ser Asn Arg Phe Ser

[0021] SEQ ID NO:6

[0022] Leu Gln Gly Thr His Gln Pro Tyr Thr

[0023] In some embodiments, the anti-α4β7 antibody comprises a heavy chain variable region (VH) having an amino acid sequence of SEQ ID NO:7 and a light chain variable region (VL) having an amino acid sequence of SEQ ID NO:8.

[0024] SEQ ID NO:7:

[0025] Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala SerVal Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His TrpVal Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile Gly Glu Ile Asp Pro Ser GluSer Asn Thr Asn Tyr Asn Gln Lys Phe Lys Gly Arg Val Thr Leu Thr Val Asp IleSer Ala Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala ValTyr Tyr Cys Ala Arg Gly Gly Tyr Asp Gly Trp Asp Tyr Ala Ile Asp Tyr Trp GlyGln Gly Thr Leu Val Thr Val Ser Ser

[0026] SEQ ID NO:8:

[0027] Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly GluPro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Ala Lys Ser Tyr Gly Asn ThrTyr Leu Ser Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr GlyIle Ser Asn Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Gly Ser Ser Gly ThrAsp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr CysLeu Gln Gly Thr His Gln Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

[0028] In some embodiments, the anti-α4β7 antibody comprises a heavy chain with an amino acid sequence of SEQ ID NO:9 and a light chain with an amino acid sequence of SEQ ID NO:10.

[0029] SEQ ID NO:9

[0030] Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala SerVal Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His TrpVal Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile Gly Glu Ile Asp Pro Ser GluSer Asn Thr Asn Tyr Asn Gln Lys Phe Lys Gly Arg Val Thr Leu Thr Val Asp IleSer Ala Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala ValTyr Tyr Cys Ala Arg Gly Gly Tyr Asp Gly Trp Asp Tyr Ala Ile Asp Tyr Trp GlyGln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe ProLeu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu ValLys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr SerGly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser SerVal Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val AsnHis Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp LysThr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val PheLeu Phe Pro Pro Lys Pro Lys AspThr Leu Met Ile Ser Arg Thr Pro Glu Val ThrCys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr ValAsp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn SerThr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly LysGlu Tyr Lys Cys Lys Val SerAsn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr IleSer Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser ArgAsp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr ProSer Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys ThrThr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr ValAsp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu AlaLeu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

[0031] SEQ ID NO:10

[0032] Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly GluPro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Ala Lys Ser Tyr Gly Asn ThrTyr Leu Ser Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr GlyIle Ser Asn Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Gly Ser Ser Gly ThrAsp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr CysLeu Gln Gly Thr His Gln Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile LysArg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu LysSer Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala LysVal Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val ThrGlu GlnAsp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser LysAla Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu SerSer Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

[0033] In some embodiments, the anti-α4β7 antibody is vedolizumab.

[0034] In some embodiments, the concentration of the anti-α4β7 antibody is about 50-70 g / L.

[0035] In some embodiments, the concentration of the anti-α4β7 antibody is about 60 g / L.

[0036] In some embodiments, the pharmaceutical formulation further comprises a second stabilizer, wherein the second stabilizer is selected from arginine hydrochloride, glycine, and sodium chloride.

[0037] In some embodiments, the second stabilizer is arginine hydrochloride.

[0038] In some embodiments, the concentration of the second stabilizer is about 22 g / L-30 g / L.

[0039] In some embodiments, the concentration of the second stabilizer is about 26.34 g / L-30 g / L.

[0040] In some embodiments, the concentration of the second stabilizer is about 26.34 g / L.

[0041] In some embodiments, the pharmaceutical formulation further comprises a surfactant, wherein the surfactant is selected from Tween 80, Tween 20, and Poloxamer 188.

[0042] In some embodiments, the surfactant is Tween 80.

[0043] In some embodiments, the concentration of the surfactant is about 0.4-0.8 g / L.

[0044] In some embodiments, the concentration of the surfactant is about 0.6 g / L.

[0045] In some embodiments, the pharmaceutical formulation further comprises a buffer, wherein the buffer is selected from the group consisting of histidine-histidine HCl and citric acid-sodium citrate.

[0046] In some embodiments, the buffer is histidine-histidine HCl.

[0047] In some embodiments, the concentration of the buffer is about 40 mM to about 65 mM.

[0048] In some embodiments, the concentration of the buffer is about 50 mM.

[0049] In some embodiments, the pharmaceutical formulation comprises an anti-α4β7 antibody, trehalose, arginine hydrochloride, Tween 80, and histidine-histidine hydrochloride buffer.

[0050] In some embodiments, the pharmaceutical formulation comprises about 50 g / L-70 g / L anti-α4β7 antibody, about 99 g / L-112 g / L trehalose, about 22 g / L-30 g / L arginine hydrochloride, about 0.4-0.8 g / L Tween 80, and about 40-65 mM histidine-histidine hydrochloride buffer.

[0051] In some embodiments, the pharmaceutical formulation comprises about 60 g / L of an anti-α4β7 antibody, about 112 g / L trehalose, about 26.34 g / L arginine hydrochloride, about 0.6 g / L Tween 80, and about 50 mM histidine-histidine hydrochloride buffer.

[0052] In some embodiments, the pH of the pharmaceutical formulation is about 6.0 to about 6.6.

[0053] In some embodiments, the pH is 6.3.

[0054] In some embodiments, the pharmaceutical preparation is a liquid preparation or a lyophilized preparation.

[0055] In some embodiments, the pharmaceutical formulation is a lyophilized formulation.

[0056] The present invention provides an optimized antibody pharmaceutical preparation for anti-α4β7 antibodies. The pharmaceutical preparation provided by the present invention has high stability when containing a high concentration of anti-α4β7 antibodies, and can maintain considerable stability when stored as a liquid preparation or a lyophilized preparation, and has low viscosity and an osmotic pressure close to isotonicity, thereby meeting the requirements for drug administration and improving the efficacy of the drug.

[0057] Definition and detailed description

[0058] The following examples further illustrate the present invention, but do not limit the present invention.

[0059] The term "antibody" herein refers to the whole antibody and any antigen-binding portion or single chain thereof. A naturally occurring "antibody" is a glycoprotein comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region comprises one domain, CL. The VH and VL regions can be further subdivided into multiple hypervariable regions, referred to as hypervariable regions or complementary determining regions (CDRs), with regions more conservatively referred to as framework regions (FRs) in between. VH and VL contain three CDRs and four FRs, arranged from amino terminus to carboxyl terminus as follows: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4, i.e., VH contains HCDR1, HCDR2, HCDR3, HFR1, HFR2, HFR3 and HFR4, and VL contains LCDR1, LCDR2, LCDR3, LFR1, LFR2, LFR3 and LFR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens.

[0060] The cell surface molecule "α4β7 integrin" or "α4β7" is a heterodimer of an α4 chain (CD49D, ITGA4) and a β7 chain (ITGB7). The human α4 and β7 genes (GenBank (National Center for Biotechnology Information, Bethesda, MD) RefSeq accession numbers NM_000885 and NM_000889, respectively) are expressed by B and T lymphocytes, particularly memory CD4+ lymphocytes. As typical of many integrins, α4β7 can exist in either a quiescent or activated state. Ligands for α4β7 include vascular cell adhesion molecule (VCAM), fibronectin, and mucosal addressin (MAdCAM (e.g., MAdCAM-1)).

[0061] The term "anti-α4β7 antibody" means a human antibody or an antigen-binding fragment thereof that specifically binds to human α4β7.

[0062] As used herein, "liquid formulations", also called liquid pharmaceutical compositions, refer to aqueous compositions that are not reconstituted from lyophilisates, which contain at least one anti-α4β7 antibody or antigen-binding fragment thereof (e.g., vedolizumab) and at least one excipient (e.g., stabilizer, etc.). The liquid pharmaceutical composition may contain other excipients (buffers, surfactants) and other active ingredients. Such formulations are also referred to as "ready-to-use" liquid formulations.

[0063] The term "stable formulation" refers to a formulation in which the antibody maintains its physical stability and / or chemical stability and / or biological activity during storage. Various analytical methods can be used to assess the stability of a given composition, for example, CEX-HPLC can be used to assess the purity of the antibody in the liquid composition of the present disclosure (the content of the main peak), and SEC-HPLC can be used to assess the aggregation level in the liquid composition of the present disclosure.

[0064] Compositions or methods described herein as "comprising" or "including" one or more named elements or steps are open ended, meaning that the named elements or steps are essential, but other elements or steps may be added within the scope of the composition or method. In any composition or method disclosed herein, known or published equivalents of any named essential element or step may be substituted for the element or step. DETAILED DESCRIPTION

[0065] The experimental methods in the following examples are conventional methods unless otherwise specified. The drug raw materials, reagent materials, etc. used in the following examples are commercially available products or can be prepared according to the known techniques in the art unless otherwise specified.

[0066] This article uses size exclusion chromatography SEC-HPLC, which can be used to evaluate the physical stability of anti-α4β7 antibodies (such as vedolizumab). SEC is used to separate monomeric anti-α4β7 antibodies (including vedolizumab) from aggregates and fragments based on size under non-denaturing conditions. "SEC purity" is used in this article to represent the percentage of monomers in SEC, which is used to evaluate the stability of anti-α4β7 antibodies (including vedolizumab). The sum of the peaks eluted before the main peak is reported as the percentage of aggregated products (aggregates, AP-SEC), and the sum of the peaks eluted after the main peak is reported as the percentage of degradation products (fragments, DP-SEC). According to the general rules of the three parts of the Chinese Pharmacopoeia 2015 edition <0512> Determination. The specific steps of the present invention are: SEC-HPLC detection is performed using a high performance liquid chromatography instrument E2695-2489 from Waters, the mobile phase is 25% methanol and 150 mM sodium phosphate solution, the detection wavelength is 280 nm, the test sample is diluted to a concentration of about 2 mg / mL antibody as the test sample solution, 20 μL of the test sample solution is injected into the liquid chromatograph, and the relative content of the monomer is reported as a numerical value, the relative content of the polymer is reported as a numerical value, and the content of the fragment is reported as a numerical value.

[0067] In this paper, cation exchange high performance liquid chromatography CEX-HPLC can be used to evaluate the chemical stability of anti-α4β7 antibodies (including vedolizumab). CEX is used to evaluate the charge heterogeneity of vedolizumab by measuring the percentage of acidic and basic variants (refer to Appendix IIIB of Part III of the 2015 edition of the Chinese Pharmacopoeia). The specific steps are: use Shimadzu high performance liquid chromatograph, Thermo Fisher ProPac TM WCX-10 chromatographic column, 20mM MES as mobile phase, wavelength 280nm, sample diluted to a concentration of 2mg / ml, then detected. Calculated by area normalization method.

[0068] Sodium dodecyl sulfate-capillary gel electrophoresis (CE-SDS) is used to evaluate the purity of anti-α4β7 antibodies (such as vedolizumab) (refer to the third edition of the Chinese Pharmacopoeia 2015). In the present invention, the CESI 8000Plus capillary electrophoresis instrument of the SCIEX manufacturer was used for detection, the test sample was diluted to 10g / L, the sample volume was 20ul, and the area normalization method was used for calculation.

[0069] The vedolizumab used in the embodiment is a self-made vedolizumab in the laboratory using conventional biotechnology, and the sequence (the amino acid sequence of the heavy chain variable region is SEQ ID NO:7, and the amino acid sequence of the light chain variable region is SEQ ID NO:8) is consistent with the sequence of vedolizumab sold on the market. The method for preparing vedolizumab is: using DNA recombination technology to construct CHO (Chinese hamster ovary) cells expressing anti-α4β7 antibodies: cloning the α4β7 coding sequence into a eukaryotic expression vector to obtain a recombinant expression vector; the obtained recombinant expression vector is transferred into CHO-GS cells, and a cell strain expressing the anti-α4β7 antibody is obtained after screening and identification. The CHO-GS cells (Chinese hamster ovary cells) in the embodiment of the present invention are purchased from Merck; the amino acid sequences of the heavy chain and light chain of the anti-α4β7 antibody are SEQ ID NO:9 and SEQ ID NO:10, respectively. CHO cells expressing the anti-α4β7 antibody are inoculated into a basal culture medium and fermented and cultured for 12-18 days, and the anti-α4β7 antibody is obtained by separation and purification using conventional biotechnology. Anti-α4β7 antibody products with the same physicochemical properties.

[0070] Vedolizumab is prepared by the conventional recombinant protein technology known above, and then the active pharmaceutical ingredient Vedolizumab is prepared with different excipients by the following preparation method:

[0071] Preparation method of replacement buffer:

[0072] Replacement buffer A: Weigh the buffer, the first stabilizer and the second stabilizer according to the prescribed amount, adjust the total volume to 7000 ml with water for injection, stir and mix evenly, and the pH is about 6.0-6.6. This is the final replacement buffer.

[0073] Preparation method of liquid preparation and lyophilized preparation:

[0074] 1. The purified sample (the active ingredient is vedolizumab) is ultrafiltered at a rate of 20 to 30 ml / min and a pressure of less than 2 bar.

[0075] 2. After ultrafiltration is completed, the protein concentration is measured. When the concentration is 40 mg / ml, the liquid is replaced in equal proportion. During the replacement operation, the conductivity and pH of the liquid at the permeate end are measured. When the pH and osmotic pressure measured at the permeate end are consistent with those of the replacement buffer, the liquid replacement is stopped.

[0076] 3. After the replacement buffer, the sample (active ingredient is vedolizumab) was ultrafiltered and concentrated to 100-120 mg / ml, the sample was collected and the volume was recorded. Then, the system was flushed with the replacement buffer and added to the over-concentrated sample to make the sample concentration 60 mg / ml.

[0077] 4. A surfactant was added to the 60 mg / ml sample (the active ingredient was vedolizumab) to make the final concentration of the surfactant 0.02%, thereby obtaining the final preparation.

[0078] 5. Filter the final preparation with a 0.22 μm filter membrane under a sterile environment and dispense into prefilled syringes at a volume of 1 ml / vial, for a total of 1,000 vials. Label them to obtain a liquid preparation.

[0079] Alternatively, the final preparation is terminally sterilized and canned into vials (5.50 ml / vial), a total of 100 vials, and then freeze-dried, labeled, and packaged into the final outer packaging material to obtain a freeze-dried preparation.

[0080] Example

[0081] Preparation

[0082] According to the above-mentioned liquid preparation method, the following preparation is obtained:

[0083] Example 1: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH adjusted to 6.3 with hydrochloric acid or sodium hydroxide, liquid preparation.

[0084] Example 2: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, adjusted to pH 6.0 with hydrochloric acid or sodium hydroxide, liquid preparation.

[0085] Example 3: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH adjusted to 6.2 with hydrochloric acid or sodium hydroxide, liquid preparation.

[0086] Example 4: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, adjusted to pH 6.6 with hydrochloric acid or sodium hydroxide, liquid preparation.

[0087] According to the above freeze-dried preparation method, the following preparation is obtained:

[0088] Example 5: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH adjusted to 6.3 with hydrochloric acid or sodium hydroxide, lyophilized preparation, the above is the concentration of the lyophilized preparation after reconstitution.

[0089] Comparative Examples 1-14 Preparation

[0090] The liquid preparations were prepared according to the above-mentioned liquid preparation preparation method to obtain the following comparative liquid preparations:

[0091] Comparative Example 1: vedolizumab 60 mg / ml, sucrose 100 mg / ml, arginine hydrochloride 26.34 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0092] Comparative Example 2: vedolizumab 60 mg / ml, sucrose 100 mg / ml, glycine 17.02 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0093] Comparative Example 3: 60 mg / ml of vedolizumab, 100 mg / ml of sucrose, 6.95 mg / ml of sodium chloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, Tween 80

[0094] 0.6mg / ml, pH 6.3, liquid formulation.

[0095] Comparative Example 4: vedolizumab 60 mg / ml, sorbitol 59 mg / ml, arginine hydrochloride 26.34 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0096] Comparative Example 5: vedolizumab 60 mg / ml, trehalose 99 mg / ml, arginine hydrochloride 30 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0097] Comparative Example 6: vedolizumab 60 mg / ml, trehalose 118 mg / ml, arginine hydrochloride 22.36 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0098] Comparative Example 7: vedolizumab 60 mg / ml, trehalose 127 mg / ml, arginine hydrochloride 18.28 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0099] Comparative Example 8: vedolizumab 60 mg / ml, trehalose 112 mg / ml, arginine hydrochloride 26.34 mg / ml, citric acid 0.8 mg / ml, sodium citrate 14.0 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0100] Comparative Example 9: vedolizumab 60 mg / ml, trehalose 112 mg / ml, arginine hydrochloride 26.34 mg / ml, glacial acetic acid 0.08 mg / ml, sodium acetate trihydrate 6.60 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0101] Comparative Example 10: vedolizumab 60 mg / ml, trehalose 112 mg / ml, arginine hydrochloride 26.34 mg / ml, sodium dihydrogen phosphate 10.5 mg / ml, sodium hydrogen phosphate 3.20 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0102] Comparative Example 11: vedolizumab 60 mg / ml, trehalose 112 mg / ml, arginine hydrochloride 26.34 mg / ml, citric acid 2.40 mg / ml, disodium hydrogen phosphate 7.00 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, liquid preparation.

[0103] Comparative Example 12: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, the pH was adjusted to 5.8 with hydrochloric acid or sodium hydroxide, liquid preparation.

[0104] Comparative Example 13: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, the pH was adjusted to 6.8 with hydrochloric acid or sodium hydroxide, liquid preparation.

[0105] Prepare according to the above freeze-dried preparation method to obtain

[0106] Comparative Example 14: vedolizumab 60 mg / ml, sucrose 100 mg / ml, arginine hydrochloride 26.34 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3, lyophilized preparation.

[0107] Test Example 1: Selection of the First Stabilizer

[0108] The effects of different first stabilizers on the physical stability (SEC-HPLC (SEC high molecular weight protein: General Rules of the Chinese Pharmacopoeia 2015 Edition) and chemical stability (CEX-HPLC lysine variants, refer to Appendix IIIB of Part III of the Chinese Pharmacopoeia 2015 Edition) of vedolizumab during storage were evaluated under long-term (4°C) and high temperature (40°C) storage conditions. The effects of the first stabilizers (sucrose, trehalose and sorbitol) were evaluated in prefilled syringes. The specific prescriptions are as follows:

[0109] Comparative Example 1: 60 mg / ml of vedolizumab, 100 mg / ml of sucrose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH 6.3.

[0110] Example 1: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH 6.3.

[0111] Comparative Example 4: vedolizumab 60 mg / ml, sorbitol 59 mg / ml, arginine hydrochloride 26.34 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 80 0.6 mg / ml, pH 6.3.

[0112] The results are shown in Tables 1-4. According to the results of accelerated and 4°C long-term stability for 24 weeks, the SEC results show that the polymer of Example 1 is lower than that of Comparative Example 1, and the CEX results show that the acidic variant of Example 1 is the lowest. In summary, trehalose as the first stabilizer has better stability than sucrose and sorbitol.

[0113] Table 1SEC-HPLC test results (40°C)

[0114]

[0115] Table 2 CEX-HPLC test results (40°C)

[0116]

[0117] Table 3SEC-HPLC test results (4°C)

[0118]

[0119]

[0120] Table 4 CEX-HPLC test results (4°C)

[0121]

[0122] Test Example 2: Selection of the Second Stabilizer

[0123] The effects of different second stabilizers on the physical stability (SEC-HPLC (SEC high molecular weight protein: General Rules of the Chinese Pharmacopoeia 2015 Edition; CE-SDS capillary electrophoresis: Part IV of the Chinese Pharmacopoeia 2020 Edition) and chemical stability (CEX-HPLC lysine variants, refer to Appendix IIIB of Part III of the Chinese Pharmacopoeia 2015 Edition) of vedolizumab during storage were evaluated under high temperature (40°C) storage conditions.

[0124] The effect of the type of stabilizer (arginine hydrochloride, glycine, sodium chloride) was evaluated in prefilled syringes. The specific formulation is as follows:

[0125] Comparative Example 1: 60 mg / ml of vedolizumab, 100 mg / ml of sucrose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH 6.3;

[0126] Comparative Example 2: 60 mg / ml of vedolizumab, 100 mg / ml of sucrose, glycine

[0127] 17.02mg / ml, histidine 4.6mg / ml, histidine hydrochloride 4.28mg / ml, Tween 80 0.6mg / ml, pH 6.3;

[0128] Comparative Example 3: 60 mg / ml of vedolizumab, 100 mg / ml of sucrose, 6.95 mg / ml of sodium chloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, Tween 80

[0129] 0.6mg / ml, pH 6.3.

[0130] The results are shown in Tables 5-7. According to the accelerated 24-week results, it can be seen from the SEC results that the polymer of Comparative Example 1 is the lowest, and from the CE results it can be seen that the purity of the antibody in Comparative Example 1 is higher, and from the CEX results it can be seen that the acid variant of Comparative Example 1 is the lowest. In summary, when arginine hydrochloride is used as the second stabilizer, the stability of the antibody preparation is better than that of glycine and sodium chloride.

[0131] Table 5 SEC-HPLC test results (40°C)

[0132]

[0133] Table 6 CE-SDS test results (40℃)

[0134]

[0135] Table 7 CEX-HPLC test results (40°C)

[0136]

[0137] Test Example 3: Selection of the content of the first stabilizer and the second stabilizer

[0138] The effects of different contents of the first stabilizer and the second stabilizer on the physical stability (SEC-HPLC, SEC high molecular weight protein: General Rules of the Chinese Pharmacopoeia 2015 Edition) and chemical stability (CEX-HPLC lysine variants, refer to Appendix IIIB of Part III of the Chinese Pharmacopoeia 2015 Edition) of vedolizumab during storage were evaluated under long-term (4°C) and high temperature (40°C) storage conditions. The effects of the stabilizer content were evaluated in prefilled syringes, and the specific formula is as follows: Comparative Example 5: vedolizumab 60 mg / ml, trehalose 99 mg / ml, arginine hydrochloride 30 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 800.6 mg / ml, pH 6.3

[0139] Example 2: 60 mg / ml of vedolizumab, 110 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH 6.3

[0140] Comparative Example 6: 60 mg / ml of vedolizumab, 118 mg / ml of trehalose, 22.36 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH 6.3

[0141] Comparative Example 7: 60 mg / ml of vedolizumab, 127 mg / ml of trehalose, 18.28 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH 6.3

[0142] The results are shown in Tables 8-11. According to the accelerated 24-week results, it can be seen from the SEC results that Example 2 (trehalose concentration is 110 mg / ml, arginine hydrochloride concentration is 26.34 mg / ml) has the lowest aggregates, and from the CEX results, it can be seen that the acidic variant of Example 2 (trehalose concentration is 110 mg / ml, arginine hydrochloride concentration is 26.34 mg / ml) is not much different from other prescriptions; and from the 4°C, 24-week results, it can be seen that Example 2 (trehalose is 110 mg / ml) and the comparative The SEC aggregate and acid peak results of Example 5 (trehalose is 99 mg / ml) are comparable and both are superior; in summary, the antibody preparation has superior stability when the trehalose concentration is about 99 mg / L-about 118 mg / mL, preferably when the trehalose concentration is about 99 mg / L-about 112 mg / mL, and more preferably when the trehalose concentration is about 110-112 mg / mL; the arginine hydrochloride has superior stability when it is about 22 mg / ml-about 30 mg / ml, preferably about 26.34 mg / ml-about 30 mg / ml, and more preferably about 26.34 mg / ml; wherein the antibody preparation has the best stability when the trehalose concentration is about 110-112 mg / ml and the arginine hydrochloride concentration is 26.34 mg / ml.

[0143] Table 8 SEC-HPLC test results (40°C)

[0144]

[0145] Table 9 CEX-HPLC test results (40°C)

[0146]

[0147]

[0148] Table 10 SEC-HPLC test results (4°C)

[0149]

[0150] Table 11 CEX-HPLC test results (4°C)

[0151]

[0152] Experimental Example 4: Selection of buffer

[0153] The effects of different buffers on the physical stability (SEC-HPLC (SEC high molecular weight protein: General Rules of the Chinese Pharmacopoeia 2015 Edition) and chemical stability (CEX-HPLC lysine variants, refer to Appendix IIIB of Part III of the Chinese Pharmacopoeia 2015 Edition) of vedolizumab during storage were evaluated under long-term (4°C) and high temperature (40°C) storage conditions. The effects of different buffers (histidine, citric acid, acetic acid, phosphoric acid, etc.) were evaluated in prefilled syringes. The specific prescription is as follows: Example 1: vedolizumab 60 mg / ml, trehalose 112 mg / ml, arginine hydrochloride 26.34 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 800.6 mg / ml, pH 6.3

[0154] Comparative Example 8: vedolizumab 60 mg / ml, trehalose 112 mg / ml, arginine hydrochloride 26.34 mg / ml, citric acid 0.8 mg / ml, sodium citrate 14.0 mg / ml, Tween 80 0.6 mg / ml, pH 6.3

[0155] Comparative Example 9: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 0.08 mg / ml of glacial acetic acid, 6.60 mg / ml of sodium acetate trihydrate, 0.6 mg / ml of Tween 80, pH 6.3

[0156] Comparative Example 10: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 10.5 mg / ml of sodium dihydrogen phosphate, 3.20 mg / ml of sodium dihydrogen phosphate, 0.6 mg / ml of Tween 80, pH 6.3

[0157] Comparative Example 11: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 2.40 mg / ml of citric acid, 7.00 mg / ml of disodium hydrogen phosphate, 0.6 mg / ml of Tween 80, pH 6.3

[0158] The results are shown in Tables 12-15. According to the results of the accelerated (40°C) and 8-week tests, it can be seen from the SEC results that Example 1 (buffer is histidine) has the lowest aggregate and the highest purity, and from the CEX results it can be seen that the acidic variant of Example 1 (buffer is histidine) has the lowest aggregate and the highest purity; followed by Comparative Example 8 (i.e., citric acid buffer); According to the long-term (4°C) and 8-week test results, it can be seen from the SEC results that Example 1 (buffer is histidine) has the lowest aggregate and the highest purity, and from the CEX results it can be seen that the acidic variant of Example 1 (buffer is histidine) is comparable to other comparative examples, and the purity is relatively high; Based on the above, the stability of the antibody preparations in Example 1 (i.e., histidine buffer) and Comparative Example 8 (i.e., citric acid buffer) is better, among which the stability of the antibody preparation in Example 1 is the best.

[0159] Table 12 SEC-HPLC test results (40°C)

[0160]

[0161]

[0162] Table 13 CEX-HPLC test results (40°C)

[0163]

[0164] Table 14 SEC-HPLC test results (4°C)

[0165]

[0166] Table 15 CEX-HPLC test results (4°C)

[0167]

[0168] Test Example 5: Effect of different pH values

[0169] Use hydrochloric acid or sodium hydroxide to adjust different pH values, and store under long-term (4°C) and high-temperature (40°C) conditions to determine the effects of storage on the physical stability (SEC-HPLC, SEC high molecular weight protein: General Rules of the 2015 edition of the Chinese Pharmacopoeia) and chemical stability (CEX-HPLC lysine variants, refer to Appendix IIIB of Part III of the 2015 edition of the Chinese Pharmacopoeia). The effect of pH on antibody stability was evaluated in prefilled syringes. The specific prescription is as follows:

[0170] Comparative Example 12: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, and the pH was adjusted to 5.8 with hydrochloric acid or sodium hydroxide.

[0171] Example 2: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, and the pH value was adjusted to 6.0 with hydrochloric acid or sodium hydroxide.

[0172] Example 3: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, and the pH value was adjusted to 6.2 with hydrochloric acid or sodium hydroxide.

[0173] Example 4: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, and the pH value was adjusted to 6.6 with hydrochloric acid or sodium hydroxide.

[0174] Comparative Example 13: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, and the pH was adjusted to 6.8 with hydrochloric acid or sodium hydroxide.

[0175] The results are shown in Tables 16-19. According to the accelerated (40°C) and long-term (4°C) results for 12 weeks, the SEC results show that when the pH is 6.0-6.6, the aggregates are lower and the purity is higher, and the CEX results show that when the pH is 6.0-6.6, the purity is the highest; based on the above, the stability is better when the pH is 6.0-6.6.

[0176] Table 16 SEC-HPLC test results (40°C)

[0177]

[0178] Table 17 CEX-HPLC test results (40°C)

[0179]

[0180]

[0181] Table 18 SEC-HPLC test results (4°C)

[0182]

[0183] Table 19 CEX-HPLC test results (4°C)

[0184]

[0185]

[0186] Experimental Example 6: Effect of different first stabilizers in freeze-dried preparations

[0187] The effects of different first stabilizers on the physical stability (SEC-HPLC (SEC high molecular weight protein: General Rules of the Chinese Pharmacopoeia 2015 Edition) and chemical stability (CEX-HPLC lysine variants, refer to Appendix IIIB of Part III of the Chinese Pharmacopoeia 2015 Edition) of vedolizumab during storage were evaluated under long-term (4°C) and high temperature (40°C) storage conditions. The stability of the formulations containing different stabilizers (sucrose and trehalose) was evaluated after lyophilization in vials. The specific prescription is as follows: Comparative Example 14: vedolizumab 60 mg / ml, sucrose 100 mg / ml, arginine hydrochloride 26.34 mg / ml, histidine 4.6 mg / ml, histidine hydrochloride 4.28 mg / ml, Tween 800.6 mg / ml, pH 6.3

[0188] Example 5: 60 mg / ml of vedolizumab, 112 mg / ml of trehalose, 26.34 mg / ml of arginine hydrochloride, 4.6 mg / ml of histidine, 4.28 mg / ml of histidine hydrochloride, 0.6 mg / ml of Tween 80, pH 6.3

[0189] The results are shown in Tables 20-23. According to the accelerated (40°C) and long-term (4°C) and 24-week results, it can be seen from the SEC results that the aggregates generated during storage in Example 5 are lower than those in Comparative Example 14, and it can be seen from the CEX results that the acidic variants generated during storage in Example 5 are also the lowest. Based on the above, the stability of the lyophilized antibody preparation of Example 5 is better, that is, when trehalose is used as the first stabilizer, the stability is better.

[0190] Table 20 SEC-HPLC test results (40°C)

[0191]

[0192]

[0193] Table 21 CEX-HPLC test results (40°C)

[0194]

[0195] Table 22 SEC-HPLC test results (4°C)

[0196]

[0197] Table 23 CEX-HPLC test results (4°C)

[0198]

[0199] The present invention has been described by the above embodiments, but it should be understood that the above embodiments are only for the purpose of example and description, and are not intended to limit the present invention to the scope of the described embodiments. In addition, it can be understood by those skilled in the art that the present invention is not limited to the above embodiments, and more variations and modifications can be made according to the teachings of the present invention, and these variations and modifications all fall within the scope of the protection claimed by the present invention. The protection scope of the present invention is defined by the attached claims and their equivalents.

Claims

1. A stable pharmaceutical preparation comprising an anti-α4β7 antibody and a first stabilizer, wherein the first stabilizer is trehalose; preferably, the concentration of the trehalose is about 99-118 g / L, more preferably, the concentration of the trehalose is about 99-112 g / L, and most preferably, the concentration of the trehalose is about 112 g / L.

2. The pharmaceutical preparation according to claim 1, wherein The anti-α4β7 antibody comprises a heavy chain variable region (VH) and, the anti-α4β7 antibody comprises a light chain variable region (VL); wherein the heavy chain variable region comprises HCDR1 with an amino acid sequence of SEQ ID NO: 1, HCDR2 with an amino acid sequence of SEQ ID NO: 2 and / or HCDR3 with an amino acid sequence of SEQ ID NO: 3; the light chain variable region comprises LCDR1 with an amino acid sequence of SEQ ID NO: 4, LCDR2 with an amino acid sequence of SEQ ID NO: 5 and / or LCDR3 with an amino acid sequence of SEQ ID NO: 6; preferably, the anti-α4β7 antibody comprises a heavy chain variable region (VH) with an amino acid sequence of SEQ ID NO: 7 and a light chain variable region (VL) with an amino acid sequence of SEQ ID NO: 8; more preferably, the anti-α4β7 antibody comprises a heavy chain with an amino acid sequence of SEQ ID NO: 9 and a light chain with an amino acid sequence of SEQ ID NO: 10; most preferably, the anti-α4β7 antibody is vedolizumab; Further preferably, the concentration of the anti-α4β7 antibody is about 50-70 g / L; even more preferably, the concentration of the anti-α4β7 antibody is about 60 g / L.

3. The pharmaceutical preparation according to any one of claims 1 to 2, wherein The pharmaceutical preparation further comprises a second stabilizer, wherein the second stabilizer is selected from arginine hydrochloride, glycine and sodium chloride; preferably, the second stabilizer is arginine hydrochloride.

4. The pharmaceutical preparation according to claim 3, wherein The concentration of the second stabilizer is about 22 g / L-30 g / L, preferably the concentration of the second stabilizer is about 26.34 g / L- 30 g / L, more preferably the concentration of the second stabilizer is about 26.34 g / L.

5. The pharmaceutical formulation according to any one of claims 1 to 4, further comprising a surfactant, wherein the surfactant is selected from Tween 80, Tween 20 and Poloxamer 188; preferably, the surfactant is Tween 80; more preferably, the concentration of the surfactant is about 0.4-0.8 g / L; most preferably, the concentration of the surfactant is about 0.6 g / L; and / or The pharmaceutical preparation further comprises a buffer, wherein the buffer is selected from histidine-histidine hydrochloride and citric acid-sodium citrate; preferably, the buffer is histidine-histidine hydrochloride; more preferably, the concentration of the buffer is about 40mM-65mM; most preferably, the concentration of the buffer is about 50mM.

6. The pharmaceutical preparation according to any one of claims 1 to 5, wherein The pharmaceutical preparation comprises anti-α4β7 antibody, trehalose, arginine hydrochloride, Tween 80 and histidine-histidine hydrochloride buffer.

7. The pharmaceutical preparation according to any one of claims 1 to 6, wherein The pharmaceutical preparation comprises about 50-70 g / L anti-α4β7 antibody, about 99-112 g / L trehalose, about 22 g / L-30 g / L arginine hydrochloride, about 0.4-0.8 g / L Tween 80 and about 40-65 mM histidine-histidine hydrochloride buffer.

8. The pharmaceutical preparation according to any one of claims 1 to 7, comprising about 60 g / L anti-α4β7 antibody, about 112 g / L trehalose, about 26.34 g / L arginine hydrochloride, about 0.6 g / L Tween 80 and about 50 mM histidine-histidine hydrochloride buffer.

9. The pharmaceutical preparation according to any one of claims 1 to 8, wherein The pH value of the pharmaceutical preparation is about 6.0-6.6, preferably the pH value is 6.

3.

10. The pharmaceutical preparation according to any one of claims 1 to 9, wherein the preparation is a liquid preparation or a lyophilized preparation.

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