Antibody formulation for coagulation factor xi and / or activated form factor xia

Abstract

The present invention relates to the field of pharmaceutical formulations, and in particular, to a liquid pharmaceutical formulation comprising an anti-FXI / FXIa antibody and an antigen-binding fragment thereof, and use thereof as a medicament.

Description

Antibody preparation against coagulation factor XI and / or its activated form factor XIa TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and relates to an antibody preparation against FXI and / or FXIa (anti-FXI / FXIa), which is used for preventing and / or treating conditions in which pathological thrombus formation or thrombus embolism is involved. BACKGROUND

[0002] Coagulation consists of humoral and cellular responses and is an important component of hemostasis, which is the process of stopping blood loss from a wound or injury. Pathological coagulation or thrombus formation refers to the formation of blood clots that are not part of the normal hemostatic process and can cause disease symptoms. Examples of pathological coagulation are deep vein thrombosis, vascular graft thrombosis, coronary atherosclerotic plaque thrombosis, microvascular thrombosis, and sepsis disseminated intravascular coagulation. In addition, part of the case thrombus can be released and carried away by the blood flow, blocking the blood vessels of other parts of the body such as the lungs or the brain, which is the process of embolism. Diseases in which thrombosis acts with or without embolism become thrombotic diseases.

[0003] At present, the therapeutic drugs for thrombotic diseases mainly include three types of drugs, namely, anticoagulants, antiplatelets and thrombolytics. Traditional anticoagulants, such as warfarin, heparin, low molecular weight heparin, and new drugs listed in recent years, such as FXa inhibitors (rivaroxaban, apixaban, etc.), and thrombin inhibitors (dabigatran, hirudin, etc.), have good effects on reducing thrombus formation, but all face the common deficiency that they can cause bleeding complications. For example, patients receiving anticoagulant therapy due to venous thrombosis have a risk of 7.2 major bleeding per 100 people per year, and the risk of fatal bleeding is 1.31 per 100 people per year, and the mortality rate due to major bleeding is 13.4% (Linkins LA, Ann Intern Med 2003; 139:893-900). Although the emergence of new oral anticoagulants (NOACs) such as apixaban and rivaroxaban has changed the rules for the use of anticoagulants, this drug has quickly occupied the main market of anticoagulants with a wider window and lower risk of bleeding than traditional anticoagulants such as warfarin. However, in vulnerable groups such as the elderly or patients with renal dysfunction, bleeding caused by NOACs is still an important clinical problem. More importantly, similar to warfarin, long-term use of NOACs increases the burden on the kidneys, leading to accelerated damage to kidney function.

[0004] Human coagulation factor XI (FXI) is an important coagulation factor in the intrinsic coagulation pathway. Research data and clinical trials show that, by inhibiting the level of FXI to 20% through antisense nucleotide drugs, an anti-coagulation effect comparable to NOACs can be achieved with a low risk of bleeding. Therefore, there is a need for a liquid pharmaceutical preparation comprising an anti-FXI and / or FXIa antibody (anti-FXI / FXIa antibody) that is sufficiently stable and suitable for administration to a patient. SUMMARY

[0005] The first aspect of the present application discloses a liquid pharmaceutical preparation comprising an anti-FXI / FXIa antibody or an antigen-binding fragment thereof and a buffer, wherein;

[0006] The anti-FXI / FXIa antibody or the antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein:

[0007] The heavy chain variable region comprises

[0008] (i) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2 and HCDR3 contained within a peptide segment having the amino acid sequence of SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81 or 83; or

[0009] (ii) a sequence comprising collectively at least one and not more than 5 amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) on the three heavy chain complementarity determining regions (HCDRs) relative to the sequence of the three heavy chain complementarity determining regions (HCDRs) recited in (i); and / or

[0010] The light chain variable region comprises,

[0011] (i) three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2 and LCDR3 contained within a peptide segment having the amino acid sequence of SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82 or 84; or

[0012] (ii) a sequence comprising collectively at least one and not more than 5 amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) on the three light chain complementarity determining regions (LCDRs) relative to the sequence of the three light chain complementarity determining regions (LCDRs) recited in (i).

[0013] In some embodiments, the anti-FXI / FXIa antibody or the antigen-binding fragment thereof comprises:

[0014] three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2 and HCDR3 contained within a peptide stretch of amino acid sequence SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81 or 83; and / or

[0015] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2 and LCDR3 contained within a peptide stretch of amino acid sequence SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82 or 84.

[0016] In some embodiments, the liquid pharmaceutical formulation comprises: an anti-FXI / FXIa antibody or antigen binding fragment thereof and a buffer, wherein the anti-FXI / FXIa antibody or antigen binding fragment thereof comprises:

[0017] a) a heavy chain complementarity determining region HCDR3 comprising or consisting of any one of the group of amino acid sequences of SEQ ID NO: 5, 1 1, 17, 23, 29, 35, 41 and 47, or comprising an amino acid sequence having 1, 2 or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as set forth in SEQ ID NO: 5, 1 1, 17, 23, 29, 35, 41 or 47; and

[0018] b) a light chain complementarity determining region LCDR3 comprising or consisting of any one of the group of amino acid sequences of SEQ ID NO: 8, 14, 20, 26, 32, 38, 44 and 50, or comprising an amino acid sequence having 1, 2 or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as set forth in SEQ ID NO: 8, 14, 20, 26, 32, 38, 44 or 50.

[0019] In some embodiments, the anti-FXI / FXIa antibody or antigen binding fragment thereof further comprises:

[0020] a) a heavy chain complementarity determining region HCDR1 comprising or consisting of any of the group of amino acid sequences set forth in SEQ ID NOs: 3, 9, 15, 21, 27, 33, 39, and 45, or comprising an amino acid sequence having 1, 2 or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence set forth as SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, or 45;

[0021] b) a heavy chain complementarity determining region HCDR2 comprising or consisting of any of the group of amino acid sequences set forth in SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, and 46, or comprising an amino acid sequence having 1, 2 or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence set forth as SEQ ID NO: 4, 10, 16, 22, 28, 34, 40, or 46;

[0022] c) a light chain complementarity determining region LCDR1 functional region comprising or consisting of any of the group of amino acid sequences set forth in SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, and 48, or comprising an amino acid sequence having 1, 2 or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence set forth as SEQ ID NO: 6, 12, 18, 24, 30, 36, 42, or 48; and

[0023] d) a light chain complementarity determining region LCDR2 functional region comprising or consisting of any of the group of amino acid sequences set forth in SEQ ID NOs: 7, 13, 19, 25, 31, 37, 43, and 49, or comprising an amino acid sequence having 1, 2 or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence set forth as SEQ ID NO: 7, 13, 19, 25, 31, 37, 43, or 49.

[0024] In some embodiments, the liquid pharmaceutical formulation comprises: an anti-FXI / FXIa antibody or antigen binding fragment thereof comprising three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3; and three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein:

[0025] HCDR1 comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, or 45, or the HCDR1 comprises an amino acid sequence having 1, 2, or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as set forth in SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, or 45;

[0026] HCDR2 comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 4, 10, 16, 22, 28, 34, 40, or 46, or the HCDR2 comprises an amino acid sequence having 1, 2, or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as set forth in SEQ ID NO: 4, 10, 16, 22, 28, 34, 40, or 46;

[0027] HCDR3 comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 5, 11, 17, 23, 29, 35, 41, or 47, or the HCDR3 comprises an amino acid sequence having 1, 2, or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as set forth in SEQ ID NO: 5, 11, 17, 23, 29, 35, 41, or 47;

[0028] LCDR1 comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 6, 12, 18, 24, 30, 36, 42, or 48, or the LCDR1 comprises an amino acid sequence having 1, 2, or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as set forth in SEQ ID NO: 6, 12, 18, 24, 30, 36, 42, or 48;

[0029] LCDR2 comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 7, 13, 19, 25, 31, 37, 43, or 49, or the LCDR2 comprises an amino acid sequence having 1, 2, or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as set forth in SEQ ID NO: 7, 13, 19, 25, 31, 37, 43, or 49;

[0030] LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO: 8, 14, 20, 26, 32, 38, 44, or 50, or the LCDR3 comprises an amino acid sequence having 1, 2, or 3 amino acid mutations, preferably amino acid substitutions, preferably conservative substitutions, compared to the amino acid sequence as shown in SEQ ID NO: 8, 14, 20, 26, 32, 38, 44, or 50.

[0031] In some embodiments, the anti-FXI / FXIa antibody or antigen binding fragment thereof comprises:

[0032] (1) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 51, and

[0033] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 52;

[0034] (2) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 53, 67, 69, 71, 73, 75, 77, 79, or 81, and

[0035] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 54, 68, 70, 72, 74, 76, 78, 80, or 82;

[0036] (3) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 55, and

[0037] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 56;

[0038] (4) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 57 or 83, and

[0039] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide stretch having an amino acid sequence as shown in SEQ ID NO: 58 or 84.

[0040] (5) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 59, and

[0041] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 60;

[0042] (6) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 61, and

[0043] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 62;

[0044] (7) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 63, and

[0045] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 64; or

[0046] (8) three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 65, and

[0047] three light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide segment of an amino acid sequence of SEQ ID NO: 66.

[0048] In some embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment thereof comprises:

[0049] (1) a HCDR1 comprising or consisting of SEQ ID NO: 3,

[0050] a HCDR2 comprising or consisting of SEQ ID NO: 4,

[0051] a HCDR3 comprising or consisting of SEQ ID NO: 5,

[0052] a LCDR1 comprising or consisting of SEQ ID NO: 6,

[0053] LCDR2, which contains SEQ ID NO:7 or is composed of therein, and

[0054] LCDR3, which contains SEQ ID NO:8 or is composed of therein;

[0055] (2) HCDR1 containing SEQ ID NO:9 or consisting thereof,

[0056] HCDR2, which contains SEQ ID NO:10 or consists of it,

[0057] HCDR3, which contains SEQ ID NO:11 or is composed of therein,

[0058] LCDR1 containing SEQ ID NO:12 or composed thereof,

[0059] LCDR2, which contains SEQ ID NO:13 or is composed of therein, and

[0060] LCDR3 containing SEQ ID NO:14 or composed thereof;

[0061] (3) HCDR1 containing or consisting of SEQ ID NO: 15,

[0062] HCDR2, which contains SEQ ID NO:16 or is composed of therein,

[0063] HCDR3, which contains SEQ ID NO:17 or is composed of therein,

[0064] LCDR1, which contains SEQ ID NO:18 or is composed of therein,

[0065] LCDR2, which contains SEQ ID NO:19 or is composed of therein, and

[0066] LCDR3, which contains SEQ ID NO:20 or is composed of therein;

[0067] (4) HCDR1 containing or consisting of SEQ ID NO:21,

[0068] HCDR2, which contains SEQ ID NO:22 or is composed of therein,

[0069] HCDR3, which contains SEQ ID NO:23 or is composed of therein,

[0070] LCDR1 containing SEQ ID NO:24 or composed thereof,

[0071] LCDR2, which contains SEQ ID NO:25 or is composed of therein, and

[0072] LCDR3 comprising or consisting of SEQ ID NO: 26;

[0073] (5) HCDR1 comprising or consisting of SEQ ID NO: 27,

[0074] HCDR2 comprising or consisting of SEQ ID NO: 28,

[0075] HCDR3 comprising or consisting of SEQ ID NO: 29,

[0076] LCDR1 comprising or consisting of SEQ ID NO: 30,

[0077] LCDR2 comprising or consisting of SEQ ID NO: 31, and

[0078] LCDR3 comprising or consisting of SEQ ID NO: 32;

[0079] (6) HCDR1 comprising or consisting of SEQ ID NO: 33,

[0080] HCDR2 comprising or consisting of SEQ ID NO: 34,

[0081] HCDR3 comprising or consisting of SEQ ID NO: 35,

[0082] LCDR1 comprising or consisting of SEQ ID NO: 36,

[0083] LCDR2 comprising or consisting of SEQ ID NO: 37, and

[0084] LCDR3 comprising or consisting of SEQ ID NO: 38;

[0085] (7) HCDR1 comprising or consisting of SEQ ID NO: 39,

[0086] HCDR2 comprising or consisting of SEQ ID NO: 40,

[0087] HCDR3 comprising or consisting of SEQ ID NO: 41,

[0088] LCDR1 comprising or consisting of SEQ ID NO: 42,

[0089] LCDR2 comprising or consisting of SEQ ID NO: 43, and

[0090] LCDR3 comprising or consisting of SEQ ID NO: 44; or

[0091] (8) HCDR1 comprising or consisting of SEQ ID NO:45,

[0092] HCDR2 comprising or consisting of SEQ ID NO:46,

[0093] HCDR3 comprising or consisting of SEQ ID NO:47,

[0094] LCDR1 comprising or consisting of SEQ ID NO:48,

[0095] LCDR2 comprising or consisting of SEQ ID NO:49, and

[0096] LCDR3 comprising or consisting of SEQ ID NO:50.

[0097] In some embodiments, the liquid pharmaceutical formulation comprises: an anti-FXI / FXIa antibody or antigen binding fragment thereof, and a buffer, the anti-FXI / FXIa antibody or antigen binding fragment thereof comprising:

[0098] a heavy chain variable region which is at least 80% identical to any one of the amino acid sequences selected from the group consisting of SEQ ID NOs:51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, and 83; preferably, the heavy chain variable region is selected from an amino acid sequence having one or more, preferably not more than 10, more preferably not more than 5, amino acid mutations, preferably amino acid substitutions, more preferably amino acid conservative substitutions, compared to SEQ ID NO:51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, or 83, preferably the amino acid mutations do not occur in the heavy chain complementarity determining regions; preferably, the heavy chain variable region has CDRs regions identical to those of SEQ ID NO:51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, or 83, and the non-CDRs regions of the heavy chain variable region are at least 80% identical to the non-CDRs regions of SEQ ID NO:51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, or 83; preferably, the heavy chain variable region has any one of the amino acid sequences selected from the group consisting of SEQ ID NOs:51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, and 83; and / or

[0099] a light chain variable region that is at least 80% identical to any one of the group consisting of the amino acid sequences of SEQ ID NOs: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, and 84; preferably, the light chain variable region is selected from an amino acid sequence having one or more, preferably no more than 10, more preferably no more than 5, amino acid mutations, preferably amino acid substitutions, more preferably amino acid conservative substitutions, compared to SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, or 84, preferably the amino acid mutations do not occur in the light chain complementarity determining regions; preferably, the light chain variable region has CDRs regions identical to the CDRs regions of the amino acid sequence of SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, or 84, and the non-CDRs regions of the light chain variable region are at least 80% identical to the non-CDRs regions of the sequence of SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, or 84; preferably, the light chain variable region has any one of the group consisting of the amino acid sequences of SEQ ID NOs: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, and 84.

[0100] In some embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region having an amino acid sequence pair selected from any one of the group consisting of the amino acid sequence pairs of SEQ ID NO: 51 and 52, SEQ ID NO: 53 and 54, SEQ ID NO: 55 and 56, SEQ ID NO: 57 and 58, SEQ ID NO: 59 and 60, SEQ ID NO: 61 and 62, SEQ ID NO: 63 and 64, SEQ ID NO: 65 and 66, SEQ ID NO: 67 and 68, SEQ ID NO: 69 and 70, SEQ ID NO: 71 and 72, SEQ ID NO: 73 and 74, SEQ ID NO: 75 and 76, SEQ ID NO: 77 and 78, SEQ ID NO: 79 and 80, SEQ ID NO: 81 and 82, and SEQ ID NO: 83 and 84.

[0101] In some embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment thereof is SEQ ID NO: 83 and 84.

[0102] In some embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment thereof further comprises a constant region from an IgG antibody, an IgM antibody, an IgA antibody, an IgD antibody, or an IgE antibody, preferably, the constant region is from an IgGl antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody.

[0103] In some embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment thereof further comprises a heavy chain constant region comprising: SEQ ID NO: 85 or an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the sequence of SEQ ID NO: 85; or consisting of SEQ ID NO: 85, or consisting of an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the sequence of SEQ ID NO: 85; and / or

[0104] a light chain constant region comprising: SEQ ID NO: 86, or an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the sequence of SEQ ID NO: 86; or consisting of SEQ ID NO: 86, or consisting of an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the sequence of SEQ ID NO: 86.

[0105] In some embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof is a murine antibody, a chimeric antibody, a humanized antibody, or a human antibody; and / or

[0106] the antibody or antigen-binding fragment thereof is selected from the group consisting of a ScFv, a Fab, a Fab', a (Fab')2, a Fv fragment, a disulfide linked Fv (dsFv), and a diabody.

[0107] In some embodiments, the liquid pharmaceutical formulation, wherein,

[0108] The buffer is selected from one or more of acetic acid, acetate, histidine, histidine hydrochloride, citric acid, citrate, phosphoric acid, phosphate, Tris (TRIS); preferably the buffer is selected from one or more of acetic acid, sodium acetate, histidine, sodium histidine, citric acid, sodium citrate, phosphoric acid, disodium hydrogen phosphate, sodium dihydrogen phosphate, TRIS; preferably the buffer is selected from one or more of acetic acid, sodium acetate, citric acid, sodium citrate, phosphoric acid, disodium hydrogen phosphate; preferably the buffer is selected from one or more of acetic acid, sodium acetate, citric acid, disodium hydrogen phosphate; more preferably the buffer is acetic acid-sodium acetate or citric acid-disodium hydrogen phosphate; more preferably acetic acid-sodium acetate.

[0109] In some embodiments, the liquid pharmaceutical formulation further comprises a surfactant and / or a stabilizer, wherein

[0110] The surfactant is selected from one or more of poloxamer 188, polysorbate 80 or polysorbate 20; preferably polysorbate 20 or polysorbate 80; preferably polysorbate 20; and / or

[0111] The stabilizer is selected from one or more of sucrose, trehalose, mannitol, sorbitol, maltose, arginine and arginine hydrochloride; preferably the stabilizer is selected from one or more of sucrose, trehalose, mannitol, sorbitol, maltose, arginine hydrochloride, preferably sorbitol or arginine hydrochloride.

[0112] In some embodiments, the liquid pharmaceutical formulation has a pH of about 5.0 to about 8.0, preferably about 5.0 to about 6.0, preferably about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9 or about 6.0.

[0113] In some embodiments, the liquid pharmaceutical formulation, wherein

[0114] the concentration of the anti-FXI / FXIa antibody or antigen binding fragment thereof is about 70 mg / mL to about 500 mg / mL, preferably about 75 mg / mL to about 500 mg / mL, preferably about 80 mg / mL to about 500 mg / mL, preferably about 100 mg / mL to about 400 mg / mL, preferably about 100 mg / ml, 110 mg / ml, 120 mg / ml, 130 mg / ml, 140 mg / ml, about 150 mg / ml, about 160 mg / ml, about 170 mg / ml, about 180 mg / ml, about 190 mg / ml, about 200 mg / ml, about 210 mg / ml, about 220 mg / ml, about 230 mg / ml, about 240 mg / ml, about 250 mg / ml, about 300 mg / ml, about 350 mg / ml, or about 400 mg / ml; and / or

[0115] the concentration of the buffer is about 1 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM; and / or

[0116] the concentration of the stabilizer is about 100 mM to about 300 mM, preferably about 100 to about 220 mM, preferably about 110 mM to about 220 mM, preferably about 120 to about 200 mM, more preferably about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 210 mM, or about 220 mM; and / or

[0117] the concentration of the surfactant is about 0.005 mg / mL to about 0.1 mg / mL, preferably about 0.005 mg / mL, about 0.006 mg / mL, about 0.007 mg / mL, about 0.008 mg / mL, about 0.009 mg / mL, about 0.01 mg / mL, about 0.02 mg / mL, about 0.03 mg / mL, about 0.04 mg / mL, about 0.05 mg / mL, about 0.06 mg / mL, about 0.07 mg / mL, about 0.08 mg / mL, about 0.09 mg / mL, or about 0.1 mg / mL.

[0118] In some embodiments, the liquid pharmaceutical formulation, comprises,

[0119] Anti-FXI / FXIa antibody or its antigen-binding fragment at a concentration of about 70 mg / mL to about 500 mg / mL, preferably about 75 mg / mL to about 500 mg / mL, preferably about 80 mg / mL to about 500 mg / mL, preferably about 100 mg / mL to about 400 mg / mL, preferably about 70 mg / mL to about 200 mg / mL, preferably about 75 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, 110 mg / mL, 120 mg / mL, 130 mg / mL, 140 mg / mL, about 150 mg / mL, about 160 mg / mL, about 170 mg / mL, about 180 mg / mL, about 190 mg / mL, or about 200 mg / mL;

[0120] The anti-FXI / FXIa antibody or its antigen-binding fragment comprises: HCDR1 comprising SEQ ID NO:21 or thereof, HCDR2 comprising SEQ ID NO:22 or thereof, HCDR3 comprising SEQ ID NO:23 or thereof, LCDR1 comprising SEQ ID NO:24 or thereof, LCDR2 comprising SEQ ID NO:25 or thereof, and LCDR3 comprising SEQ ID NO:26 or thereof; preferably, the anti-FXI / FXIa antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the amino acid sequence of HCDR1 is as shown in SEQ ID NO:21, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:22, the amino acid sequence of HCDR3 is as shown in SEQ ID NO:23, the amino acid sequence of LCDR1 is as shown in SEQ ID NO:24, the amino acid sequence of LCDR2 is as shown in SEQ ID NO:25, and the amino acid sequence of LCDR3 is as shown in SEQ ID NO:26. As shown in NO:26; preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or its antigen-binding fragment are shown in SEQ ID NO:83 and SEQ ID NO:84, respectively; and

[0121] An acetate / sodium acetate buffer solution or citrate-disodium hydrogen phosphate at a concentration of about 1 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 10 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM; and

[0122] about 150 mM to about 220 mM, more preferably about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, or about 220 mM sorbitol or arginine hydrochloride; and

[0123] about 0.005 mg / mL to about 0.1 mg / mL, preferably about 0.005 mg / mL, about 0.006 mg / mL, about 0.007 mg / mL, about 0.008 mg / mL, about 0.009 mg / mL, about 0.01 mg / mL, about 0.02 mg / mL, about 0.03 mg / mL, about 0.04 mg / mL, about 0.05 mg / mL, about 0.06 mg / mL, about 0.07 mg / mL, about 0.08 mg / mL, about 0.09 mg / mL, or about 0.1 mg / mL polysorbate 20 or polysorbate 80;

[0124] the pH is about 5.0 to about 8.0, preferably about 5.0 to about 6.0, preferably about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.0.

[0125] In some embodiments, the liquid pharmaceutical formulation comprises

[0126] a) 150 mg / ml ± 50 mg / ml or 75 mg / ml ± 20 mg / ml of an anti-FXI / FXIa antibody, or antigen binding fragment thereof; preferably about 150 mg / ml of an anti-FXI / FXIa antibody, or antigen binding fragment thereof; preferably about 75 mg / ml of an anti-FXI / FXIa antibody, or antigen binding fragment thereof;

[0127] b) 25 mM ± 5 mM of an acetic acid / sodium acetate buffer; preferably about 25 mM of an acetic acid / sodium acetate buffer;

[0128] c) 150 mM ± 10 mM, 180 mM ± 10 mM, or 220 mM ± 10 mM of sorbitol, preferably about 150 mM to about 220 mM of sorbitol; preferably about 150 mM, about 180 mM, or about 220 mM of sorbitol; and

[0129] d) 0.01 mg / ml ± 0.005 mg / ml of polysorbate 20; preferably, about 0.01 mg / ml of polysorbate 20; and

[0130] pH 5.3 ± 0.3; preferably pH is about 5.5;

[0131] Preferably, the anti-FXI / FXIa antibody, or antigen binding fragment thereof, comprises a HCDR1 comprising or consisting of SEQ ID NO: 21, a HCDR2 comprising or consisting of SEQ ID NO: 22, a HCDR3 comprising or consisting of SEQ ID NO: 23, a LCDR1 comprising or consisting of SEQ ID NO: 24, a LCDR2 comprising or consisting of SEQ ID NO: 25, and a LCDR3 comprising or consisting of SEQ ID NO: 26; preferably, the anti-FXI / FXIa antibody, or antigen binding fragment thereof, comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3, the amino acid sequence of the HCDR1 is set forth in SEQ ID NO: 21, the amino acid sequence of the HCDR2 is set forth in SEQ ID NO: 22, the amino acid sequence of the HCDR3 is set forth in SEQ ID NO: 23, the amino acid sequence of the LCDR1 is set forth in SEQ ID NO: 24, the amino acid sequence of the LCDR2 is set forth in SEQ ID NO: 25, and the amino acid sequence of the LCDR3 is set forth in SEQ ID NO: 26; preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody, or antigen binding fragment thereof, are set forth in SEQ ID NO: 83 and SEQ ID NO: 84, respectively.

[0132] In some embodiments, the liquid pharmaceutical formulation, comprises

[0133] a) 150 mg / ml ± 50 mg / ml of the anti-FXI / FXIa antibody, or antigen binding fragment thereof; preferably about 150 mg / ml of the anti-FXI / FXIa antibody, or antigen binding fragment thereof;

[0134] b) 25 mM ± 5 mM of citric acid-dibasic sodium phosphate; preferably about 25 mM of citric acid-dibasic sodium phosphate;

[0135] c) 150 mM ± 10 mM of sorbitol, preferably about 150 mM of sorbitol; and

[0136] d) 0.05 mg / ml ± 0.01 mg / ml of polysorbate 20; preferably about 0.05 mg / ml of polysorbate 20; and

[0137] pH 5.3 ± 0.3; pH is about 5.5;

[0138] Preferably, the anti-FXI / FXIa antibody or antigen-binding fragment thereof comprises a HCDR1 comprising or consisting of SEQ ID NO: 21, a HCDR2 comprising or consisting of SEQ ID NO: 22, a HCDR3 comprising or consisting of SEQ ID NO: 23, a LCDR1 comprising or consisting of SEQ ID NO: 24, a LCDR2 comprising or consisting of SEQ ID NO: 25, and a LCDR3 comprising or consisting of SEQ ID NO: 26; preferably, the anti-FXI / FXIa antibody or antigen-binding fragment thereof comprises a HCDR1 having the amino acid sequence of SEQ ID NO: 21, a HCDR2 having the amino acid sequence of SEQ ID NO: 22, a HCDR3 having the amino acid sequence of SEQ ID NO: 23, a LCDR1 having the amino acid sequence of SEQ ID NO: 24, a LCDR2 having the amino acid sequence of SEQ ID NO: 25, and a LCDR3 having the amino acid sequence of SEQ ID NO: 26; preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or antigen-binding fragment thereof are set forth in SEQ ID NO: 83 and SEQ ID NO: 84, respectively.

[0139] In some embodiments, the liquid pharmaceutical formulation, comprises

[0140] a) 150 mg / ml ± 50 mg / ml or 75 mg / ml ± 20 mg / ml of the anti-FXI / FXIa antibody or antigen-binding fragment thereof; preferably about 150 mg / ml of the anti-FXI / FXIa antibody or antigen-binding fragment thereof; preferably about 75 mg / ml of the anti-FXI / FXIa antibody or antigen-binding fragment thereof;

[0141] b) 25 mM ± 5 mM of an acetic acid / sodium acetate buffer; preferably about 25 mM of an acetic acid / sodium acetate buffer;

[0142] c) about 100 mM to about 150 mM arginine hydrochloride; preferably 110 mM ± 10 mM arginine hydrochloride; preferably about 110 mM or about 140 mM arginine hydrochloride; and

[0143] d) 0.01 mg / ml ± 0.005 mg / ml of polysorbate 20 or polysorbate 80; preferably about 0.01 mg / ml of polysorbate 20 or polysorbate 80; and

[0144] a pH of 5.5 ± 0.3; preferably a pH of about 5.5;

[0145] Preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody, or antigen binding fragment thereof, are set forth in SEQ ID NO: 83 and SEQ ID NO: 84, respectively.

[0146] In some embodiments, the liquid pharmaceutical formulation, comprises

[0147] a) about 150 mg / ml of an anti-FXI / FXIa antibody, or antigen binding fragment thereof, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody, or antigen binding fragment thereof, are set forth in SEQ ID NO: 83 and SEQ ID NO: 84, respectively;

[0148] b) about 25 mM of an acetate buffer;

[0149] c) about 110 mM of arginine hydrochloride; and

[0150] d) about 0.01 mg / ml of polysorbate 20 or polysorbate 80; and

[0151] at a pH of about 5.5.

[0152] In some embodiments, the liquid pharmaceutical formulation, comprises

[0153] a) about 150 mg / ml of an anti-FXI / FXIa antibody, or antigen binding fragment thereof, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody, or antigen binding fragment thereof, are set forth in SEQ ID NO: 83 and SEQ ID NO: 84, respectively;

[0154] b) about 25 mM of an acetate buffer;

[0155] c) about 180 mM of sorbitol; and

[0156] d) about 0.01 mg / ml of polysorbate 20 or polysorbate 80; and

[0157] at a pH of about 5.5.

[0158] In some embodiments, the liquid pharmaceutical formulation is administered to a subject by a mode of administration selected from the group consisting of subcutaneous administration, intravenous administration, inhalation administration, intradermal administration, transdermal administration, intraperitoneal administration, and intramuscular administration.

[0159] A second aspect of the present application provides a lyophilized formulation prepared from the liquid pharmaceutical formulation of the first aspect of the present application.

[0160] The third aspect of the application provides a method of preparing the liquid pharmaceutical formulation of the first aspect of the application, comprising mixing the anti-FXI / FXIa antibody or antigen-binding fragment thereof and one or more pharmaceutically acceptable excipients.

[0161] The fourth aspect of the application provides the use of the liquid pharmaceutical formulation of the first aspect of the application or the lyophilized formulation of the second aspect of the application in the manufacture of a medicament for the treatment of coagulation or thrombotic embolism.

[0162] The fifth aspect of the application provides a method of treating coagulation or thrombotic embolism, comprising administering to a subject in need thereof a therapeutically effective amount of the liquid pharmaceutical formulation of the first aspect of the application or the lyophilized formulation of the second aspect of the application.

[0163] The sixth aspect of the application provides a container comprising the liquid pharmaceutical formulation of the first aspect of the application or the lyophilized formulation of the second aspect of the application, the container being a tube, a bottle, a vial, a syringe or an injection pen.

[0164] The seventh aspect of the application provides a kit comprising the container of the sixth aspect of the application.

[0165] Definitions

[0166] For better understanding of the present application, the definitions and explanations of relevant terms are provided as follows.

[0167] The term "antibody" or "Ab" generally refers to a Y-shaped tetrameric protein comprising two heavy (H) and two light (L) polypeptide chains held together by covalent disulfide bonds and non-covalent interactions. The light chains of antibodies can be classified into kappa and lambda light chains. The heavy chains can be classified into mu, delta, gamma, alpha, and epsilon, which define the isotype of the antibody as IgM, IgD, IgG, IgA, and IgE, respectively. Antibodies also have different antibody isotypes, such as IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtypes), IgA1, IgA2, IgD, IgE, or IgM antibodies. Each heavy chain comprises a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region comprises 3 domains (CH1, CH2, and CH3). Each light chain comprises a light chain variable region (VL) and a light chain constant region (CL). The VH and VL regions can be further divided into hypervariable regions (called complementarity determining regions (CDRs)) that are spaced apart by relatively conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from N- to C-terminus. The variable regions of each heavy / light chain pair (VH and VL) form an antigen binding site, respectively.

[0168] The terms "antigen-binding portion" or "antigen-binding fragment" of an antibody, which may be used interchangeably in the context of this application, refer to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been demonstrated that the antigen-binding function of an antibody can be achieved by certain fragments of a full-length antibody. Under certain conditions, antigen-binding fragments include Fab, Fab', F(ab')2, Fd, Fv, dAb, and complementarity-determining regions.

[0169] (CDR) fragments, single-chain antibodies (e.g., scFv), chimeric antibodies, biantibodies, and such polypeptides comprising at least a portion of an antibody sufficient to confer specific antigen-binding ability. The antigen-binding fragment of the antibody can be obtained from a given antibody using conventional techniques known to those skilled in the art (e.g., recombinant DNA techniques or enzymatic or chemical cleavage methods) and can be screened for specificity in the same manner as the intact antibody.

[0170] Different analyses can be used to determine or roughly estimate the CDR region. Examples of such methods include, but are not limited to, IMGT definitions, Kabat definitions, Chothia definitions, AbM definitions, and contact definitions. The IMGT definition, or International Immunogenetic Information (IMGT) (LaFranc et al., 2005. Nucl Acids Res. 33: D593-D597), has become a widely adopted and universally accepted numbering system. The Kabat definition is an immunoglobulin alignment and numbering system proposed by Elvin A. Kabat (see, for example, Kabat et al., Sequences of Proteins of Immunological Interest, 5). thEd. Public Health Service, National Institutes of Health, Bethesda, Md., 1991. or) e.g., Johnson & Wu, Nucleic Acids Res., 28:214-8 (2000), for the standard used for numbering of residues in antibodies, generally used to determine CDR regions. The Chothia definition is similar to the Kabat definition, but the Chothia definition takes into account the location of certain structural loop regions, which is a classic rule for identifying CDR boundaries based on the location of structural loop regions using the immunoglobulin numbering system proposed by Chothia et al., see, e.g., Chothia et al., J. Mol. Biol., 196:901-17 (1986); Chothia et al., Nature, 342:877-83 (1989). The AbM definition uses integrated computer programs that mimic antibody structure made by Oxford Molecular Group. See, e.g., Martin et al., Proc Natl Acad Sci (USA), 86:9268-9272 (1989); "AbM™, A Computer Program for Modeling Variable Regions of Antibodies" Oxford, UK; Oxford Molecular, Ltd. The AbM definition models antibody tertiary structure from primary sequence using known databases and ab initio methods, such as described in Samudrala et al., "Ab Initio Protein Structure Prediction Using a Combined Hierarchical Approach" PROTEINS, Structure, Function and Genetics Suppl., 3:194-198 (1999). The contact definition is based on analysis of available complex crystal structures. See, e.g., MacCallum et al., J. Mol. Biol., 5:732-45 (1996).

[0171] The antibody or antigen-binding fragment thereof described herein can be a murine antibody, a chimeric antibody, a humanized antibody, or a human antibody.

[0172] The term "monoclonal antibody" or "monoclonal" or "mAb" refers to antibodies of single molecular composition or preparations thereof. Monoclonal antibodies display a single binding specificity and affinity to a particular epitope.

[0173] The term "murine antibody" or "murine" refers to antibodies obtained by fusing immunized mouse B cell myeloma cells, followed by screening for mouse hybridoma cells that both multiply indefinitely and secrete antibodies, followed by screening, antibody production and antibody harvesting; or, to antibodies produced by plasma cells that are formed by the differentiation and multiplication of B cells following antigen invasion into the mouse.

[0174] The term "chimeric antibody" refers to an antibody in which the variable region sequences are from one species and the constant region sequences are from another species, such as an antibody in which the variable region sequences are derived from a mouse antibody and the constant region sequences are derived from a human antibody.

[0175] The term "humanized antibody" is intended to refer to antibodies in which CDR sequences derived from another mammalian species, such as mouse, have been grafted onto human framework sequences. Additional framework region modifications can be made within the human framework sequences. The terms "FXI protein", "FXI antigen" and "FXI" are used interchangeably and refer to the coagulation factor XI protein of different species. The terms "FXIa protein", "FXIa antigen" and "FXIa" are used interchangeably and refer to the activated coagulation factor XI protein of different species. The terms "FXI" and "FXIa" include both the native FXI and FXIa proteins, mutants and variants of the native FXI and FXIa proteins (mutants and variants of the native primary structure having substantially the same amino acid sequence).

[0176] The term "therapeutically effective amount" refers to the amount of an antibody that, when administered to a subject for the treatment of at least one clinical symptom of a disease or disorder, is sufficient to effect such treatment for the disease, disorder or symptom. The therapeutically effective amount can vary as a function of the severity of the disease or disorder, or the age and / or weight of the subject to be treated.

[0177] The term "pharmaceutically acceptable excipient" or "excipient" refers to a carrier and / or excipient that is physiologically and / or pharmacologically compatible for use in a subject and an active agent, which are well known in the art. These include, but are not limited to, pH adjusting agents, surfactants, adjuvants, ionic strength enhancers, viscosity reducing agents, antioxidants, buffers, preservatives, isotonic agents, chelating agents and stabilizers, etc.

[0178] The term "pharmaceutical formulation" refers to a pharmaceutical composition comprising an active agent in combination with an inert or active carrier, which makes the composition particularly suitable for diagnostic or therapeutic use in vivo or in vitro.

[0179] The term "stable" refers to formulations in which all of the protein therein essentially retain their physical, chemical, biological activity after storage at the intended storage temperature, e.g., 0-40°C. A formulation can be stable even though the antibody in the formulation does not retain 100% of its physical, chemical, biological activity after storage for some time. A formulation can be considered "stable" if it retains about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more of the antibody structure and function after storage for some time.

[0180] The present application provides containers comprising the stable pharmaceutical formulations provided herein. The term "container" can be a tube, a bottle, a vial, a syringe, or an injection pen. In embodiments where the container is a syringe, the container further comprises a needle for injection. Accordingly, the present application provides containers comprising stable liquid formulations for single dose administration or multiple dose administration.

[0181] The term "kit" can comprise the pharmaceutical formulations described herein and a device for administration, e.g., the pharmaceutical formulation can be packaged with a device for administration, e.g., a syringe, an inhaler, a measuring cup, a dropper, or an applicator. The pharmaceutical formulation can be filled in a container as defined above. The kit can optionally comprise instructions for use, including instructions for dosage, administration regimen, and mode of administration. BRIEF DESCRIPTION OF DRAWINGS [0181.1][Corrected according to Rule 91 04.09.2025] Figures 1a-1g are the results of S2366 activity detection of chimeric antibodies 24B6, 14F3, 29E10C4, 17E3C10, 19C9C10, 11E8, 3B3 in turn, with the horizontal axis being concentration (nM) and the vertical axis being absorbance OD405; [0181.2][Corrected according to Rule 91 04.09.2025] Figures 2a-2g are the results of FXIa kit activity detection of chimeric antibodies 24B6, 14F3, 17E10, 29E10C4, 17E3C10, 19C9C10, 11E8 in turn, with the horizontal axis being concentration (nM) and the vertical axis being absorbance OD405; [0181.3][Corrected according to Rule 91 04.09.2025] Figures 3a-3h are the results of human blood APTT prolongation experiments of chimeric antibodies 24B6, 14F3, 17E10, 29E10C4, 17E3C10, 19C9C10, 11E8, 3B3 in turn, with the horizontal axis being concentration (nM) and the vertical axis being the time (sec) for which precipitation was observed; [0181.4][Corrected according to Rule 91 04.09.2025] Figure 4 is the result of S2366 activity detection of humanized antibodies 14F3-Z1, 14F3-Z2, 14F3-Z3, 14F3-Z4, 14F3-1, 14F3-2, 14F3-3, 14F3-4, the horizontal axis is the concentration (nM), and the vertical axis is the absorbance OD405; [0181.5][Corrected according to Rule 91 04.09.2025] Figure 5 is the result of S2366 activity detection of humanized antibody 29E10C4-1, the horizontal axis is the concentration (nM), and the vertical axis is the absorbance OD405; [0181.6][Corrected according to Rule 91 04.09.2025] Figure 6 is the result of FXIa kit activity detection of humanized antibodies 14F3-Z1, 14F3-Z2, 14F3-Z3, 14F3-Z4, 14F3-1, 14F3-2, 14F3-3, 14F3-4, the horizontal axis is the concentration (nM), and the vertical axis is the absorbance OD405; [0181.7][Corrected according to Rule 91 04.09.2025] Figure 7 is the result of FXIa kit activity detection of humanized antibody 29E10C4-1 and murine antibody 29E10C4-sy, the horizontal axis is the concentration (nM), and the vertical axis is the absorbance OD405; [0181.8][Corrected according to Rule 91 04.09.2025] Figure 8 is the result of human blood APTT prolongation experiment of humanized antibodies 14F3-Z1, 14F3-Z2, 14F3-Z3, 14F3-Z4, 14F3-1, 14F3-2, 14F3-3, 14F3-4, the horizontal axis is the concentration (nM), and the vertical axis is the time (sec) for observing precipitation; [0181.9][Corrected according to Rule 91 04.09.2025] Figure 9 is the result of human blood APTT prolongation experiment of humanized antibody 29E10C4-1, the horizontal axis is the concentration (nM), and the vertical axis is the time (sec) for observing precipitation; [0181.10][Corrected according to Rule 91 04.09.2025] Figure 10 is the result of free FXI concentration detection in vivo of humanized antibody 29E10C4-1 after injection into cynomolgus monkeys, the horizontal axis is the time (h / d), and the vertical axis is the relative baseline level of free FXI. [0181.11][Corrected according to Rule 91 04.09.2025] Figure 11 is the result of pharmacokinetics of humanized antibodies 14F3-1 and 29E10C4-1 in vivo in cynomolgus monkeys, the horizontal axis is the time (m / h / d), and the vertical axis is the antibody concentration in plasma (μg / mL). [0181.12][Correspondence 91 more corrections 04.09.2025] Figure 12 is the result of PT (plasma thrombin time) test of humanized antibodies 14F3-1 and 29E10C4-1 after administration in cynomolgus monkeys, the horizontal axis is time (m / h / d), and the vertical axis is PT (sec). DETAILED DESCRIPTION

[0182] The embodiments of the present application will be described in detail below with examples, but those skilled in the art will understand that the following examples are only for illustration of the present application and should not be regarded as limiting the scope of the present application. The specific conditions not mentioned in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not marked with the manufacturer, which are all conventional products that can be purchased on the market.

[0183] Abbreviations

[0184] TMB refers to 3,3',5,5'-tetramethylbenzidine

[0185] Solution.

[0186] PBS refers to phosphate buffer solution (PBS, Phosphate Buffer Solution).

[0187] PBST refers to phosphate buffer solution (PBS, Phosphate Buffer Solution) added with Tween-20 (Tween-20).

[0188] BSA refers to bovine serum albumin (BSA, Bovine Serum Albumin).

[0189] IC 50 refers to the concentration that produces 50% inhibition (half maximal inhibitory concentration).

[0190] ELISA refers to enzyme linked immunosorbent assay.

[0191] APTT is activated partial thromboplasting time.

[0192] Example 1 Preparation of antibodies

[0193] Animal immunization:

[0194] Use 1 mL medical syringe injection of 500 uL PBS pH 7.2 basic (1x) (Gibco), take 1 mg target protein plus PBS PH 7.2 basic (1x) (Gibco) to 500 μL mixed evenly, and then mixed with adjuvant (Sigma Adjuvant System, SIGMA) (freeze-dried powder adjuvant stored in 4℃ medical refrigerator was heated in 42℃ water bath for 15 min, ready for use), vortex for 5 min, and then ready for use. The mice were immunized according to the following method, 200 μg target protein, 200 μL per mouse, subcutaneous injection in abdominal cavity and back, two points were selected for each injection, 50 μL per point; the second immunization was performed 3 weeks after the first immunization, and the immunization method was the same as the first immunization; the third immunization was performed 3 weeks later, and the immunization method was the same as the second immunization, and the mouse tail vein blood was taken one week later to measure the titer. The mice with good titer were selected for fusion three days before, i.e. abdominal injection of 100 μg target protein diluted to 100 μL without adjuvant for immunization, and the boost could not add adjuvant.

[0195] The titer determination method is as follows: 10 μL of mouse tail vein blood is dissolved in 490 μL of PBS, and is diluted from 1:3000 to 240,000 by 3-fold gradient, respectively. 100 μL is taken to the Elisa plate coated with the target protein (100 ng / well), and is incubated at 37℃ in a biochemical incubator for 1 h; the residual solution is shaken off by washing 3 times with PBST as the washing solution, 100 μL of enzyme-labeled secondary antibody diluted with blocking solution (diluted at 1:10000) is added, and is incubated at 37℃ in a biochemical incubator for 1 h; the residual solution is shaken off by washing 4 times with PBST as the washing solution, 100 μL of TMB single-component color developing liquid is added, and is colored for about 8 min in the dark, 50 μL of 2M H2SO4 is added to terminate the reaction, and the OD value is read by an enzyme-labeled instrument, and the titer is P / N>2.1. 450

[0196] Hybridoma fusion:

[0197] The mouse spleen cells and Sp2 / 0 mouse myeloma cells were fused by standard electrofusion, and then screened by HAT selection medium (DMEM medium containing 20% FBS, 1% HAT, 1% P / S, 1% OPI) added with OPI, and screened by Elisa after 7 days.

[0198] Indirect Elisa operation steps:

[0199] ​On the 7th day after fusion, the medium was aspirated as much as possible without sucking out the hybridoma cells, and 180 μL of HAT selection medium (Gibco) was added in the afternoon one day before ELISA detection. 80 μL of cell supernatant was added to the detection plate coated with target protein, and a positive control (positive mouse serum, diluted 1:1000) was set. Incubate in a 37°C biochemical incubator for 1 hour. Wash 3 times with PBST to remove residual solution, add enzyme-labeled secondary antibody (Invitrogen) diluted 1:10000 with PBST+2% BSA, 100 μL per well, and incubate in a 37°C biochemical incubator for 1 hour. Wash 4 times with PBST to remove residual solution. Add TMB single-component developing solution (Solarbio), 100 μL / well, and develop in the dark for about 8 minutes. Add 2M H2SO4 stop solution to stop the reaction, 50 μL / well. Use a microplate reader to read the OD 450 value. Select positive wells and transfer them to a 24-well plate for further culture for two days before freezing.

[0200] Cell subcloning:

[0201] The hybridoma cells were plated at a density of one cell per well using limited dilution method, and the positive wells containing single cell clusters were detected by the method of Cell Base Elisa experiment. Specifically, the cells in a 12-well plate were gently blown and mixed with 20 μL of cell suspension and 20 μL of trypan blue. The number of cells was counted using a countstar, and according to the counting result, 300 cells were transferred to 40 ml of complete medium (complete medium). The complete medium reagent composition is: 20% FBS (EeCell Bio) + 1% P / S (Gibco) + 1% HAT (Gibco) + 1% OPI (SIGMA) + DMEM (Gibco). The 40 ml medium was plated at a density of 200 μL / well in two plates and incubated in a 37°C carbon dioxide incubator. On the 5th day, the cell state and the number of cell strains per well were observed under a microscope and marked. After 7 days, the positive wells were detected by indirect Elisa experiment, and the single and positive cell strains were expanded to a 24-well plate. After two days, they were expanded to a 6-well plate, and when they reached 70%-80% density, they were frozen, with two strains of each cell frozen and stored in liquid nitrogen.

[0202] Sequencing of hybridoma positive antibody

[0203] Eight FXI target-related hybridomas were sequenced and analyzed, as shown in Table 1:

[0204] Table 1 Murine antibody sequences

[0205] Detection of murine antibody activity

[0206] The frozen monoclonal cell strains are recovered, the serum content in the culture medium is gradually reduced, and finally the monoclonal cell strains and antibodies surviving in the serum-free culture medium are domesticated. The antibodies obtained by domestication are subjected to S2366 activity detection according to the following method: the FXIa factor is mixed with the antibodies, added to a 96-well plate, and then S2366 solution (BIOO) is added. After shaking and mixing, the OD value at 405 nm is read. The MAA868 antibody is used as a positive control, and the antibodies are subjected to S2366 detection. The concentration of the antibody to be tested is 20 μg / mL. The S2366 detection results of the antibodies are shown in Table 2:

[0207] Table 2 S2366 activity detection results of mouse-derived antibodies after domestication

[0208] The experimental results show that the OD values of the mouse-derived antibodies disclosed in the patent are lower than that of the positive control MAA868, and therefore the mouse-derived antibodies disclosed in the patent have better activity than the positive control drug MAA868. 405nm

[0209] Preparation of chimeric antibodies

[0210] Primers are designed according to the sequences obtained by sequencing the hybridoma, and the variable region of the mouse-derived antibody is replaced into the pcDNA3.1(+) vector containing the constant region of the human-derived antibody by enzyme digestion and ligation, to form a chimeric antibody vector of the variable region of the mouse-derived antibody and the constant region of the human-derived antibody. The vector is sequenced and correct, and is transfected into 293F cells. The supernatant expressed by the cells is collected, purified, and then the corresponding chimeric antibodies 24B6, 14F3, 17E10, 29E10C4, 17E3C10, 11E8, 19C9C10 and 3B3 are obtained. The chimeric antibodies can avoid the immunogenicity of heterologous antibodies, while retaining the ability of specific binding of antibodies to antigens.

[0211] Activity detection of chimeric antibodies

[0212] S2366 activity detection is performed according to the following method: the FXIa factor is mixed with the antibodies, added to a 96-well plate, and then S2366 solution (BIOO) is added. After shaking and mixing, the OD value at 405 nm is read. The MAA868 antibody is used as a positive control, and the antibodies are subjected to S2366 detection and IC 50 values are calculated. The antibody to be tested and MAA868 are diluted with PBS, and the initial concentration is 0.55 mg / mL. The dilution is two-fold down, and seven concentrations are detected. The S2366 detection results of the antibodies are shown in Figures 1a-1g, and the IC 50 values are shown in Table 3:

[0213] Table 3 IC 50 values of chimeric antibodies S2366 activity detection

[0214] The experimental results show that the IC 50 of the chimeric antibodies of the present disclosure are all lower than or equivalent to the positive control MAA868, and the chimeric antibodies of the present disclosure are better or equivalent to the positive control MAA868 in activity.

[0215] FXIa kit activity detection

[0216] The FXIa kit activity detection was performed according to the steps of the FXIa kit (HYPHEN BioMed, BIOPHEN TM Factor Xia) instruction manual. The MAA868 antibody was used as a positive control, and each antibody to be tested was sequentially subjected to FXIa kit detection and the IC 50 was calculated. The chimeric antibodies to be tested and MAA868 were diluted with PBS, and the initial concentration was 2.2 mg / mL, which was sequentially diluted by four times down, and eight concentrations were detected. The obtained FXIa kit detection results are shown in Figures 2a-2g, and the IC 50 values are shown in Table 4:

[0217] Table 4: IC 50

[0218] The experimental results show that the IC 50 of each antibody of the present disclosure are all lower than or equivalent to the positive control MAA868, and in particular, the antibodies 29E10C4 and 17E3C10 show IC 50 far lower than the positive control, and the antibodies to be tested are better or equivalent to the positive control MAA868 in activity.

[0219] Human blood APTT extension experiment

[0220] The anticoagulant activity of the antibodies to be tested was detected using human plasma (Osocimab) and activated partial thromboplastin time (APTT) detection kits (Regen Bio) according to the steps of the kit instruction manual. The MAA868 and Osocimab antibodies were used as positive controls, and the chimeric antibodies were subjected to APTT function detection. The chimeric antibodies to be tested and the positive control drugs were diluted with PBS, and the initial concentration was 0.8 mg / mL, which was sequentially diluted by four times down, and six concentrations were detected.

[0221] The detection results are shown in Figures 3a-3h. As can be seen from the results, with the increase of the antibody concentration, the APTT time is increased to different degrees, and the test antibody has a good prolongation of the clotting time compared with the positive control drug. The clotting time of the test antibodies 24B6, 17E10 and 17E3C10 is shorter than that of the control antibody at low concentration, and is longer than that of the control drug at high concentration; the clotting time of 14F3, 29E10C4 and 19C9C10 is close to that of the positive control drug at low concentration, and is longer than that of the positive control at high concentration; the clotting time of 11E8 and 3B3 is close to that of the positive control drug.

[0222] Humanization of chimeric antibodies

[0223] The 14F3 antibody and the 29E10C4 antibody with excellent activity screened are subjected to humanization. By using computer simulation technology, the human germ line antibody sequence is selected according to the homology of the murine antibody and the human antibody, and CDR grafting is performed. Based on the three-dimensional structure of the murine antibody, by back mutation or non-back mutation of the key amino acids in the FR region of the humanized antibody, a humanized antibody molecule with high degree of humanization and excellent activity is finally obtained.

[0224] The constant region sequence of the humanized antibody is as shown in SEQ ID NO: 85 and SEQ ID NO: 86:

[0225] The heavy chain constant region sequence is as shown in SEQ ID NO: 87:

[0226] The light chain constant region sequence is as shown in SEQ ID NO: 88:

[0227] The humanized sequence of the monoclonal antibody 14F3 is shown in Table 5:

[0228] Table 5 Humanized antibody sequence of the monoclonal antibody 14F3

[0229] The humanized sequence of the monoclonal antibody 29E10C4 is shown in Table 6:

[0230] Table 6 Humanized antibody sequence of the monoclonal antibody 29E10C4 29E10C4-1

[0231] Activity detection of humanized antibodies

[0232] Activity detection of S2366

[0233] S2366 activity detection was performed as follows: FXIa factor was mixed with antibody, added to 96-well plate, then S2366 solution (Biorad) was added, after shaking, OD value at 405 nm was read. Osocimab and MAA868 antibody were used as positive control, and each antibody was detected by S2366 and IC 50 value was calculated. The results are shown in Figures 4-5 and Tables 7-8. 50

[0234] Table 7 IC value of humanized antibody of antibody 14F3 in S2366 detection 50

[0235] The results showed that the IC 50 values of the eight humanized antibodies disclosed in the present patent were lower than the positive control, especially the IC 50 values of 14F3-1 and 14F3-2 were the lowest, 45.38 nM and 42.96 nM respectively, so the humanized antibodies disclosed in the present patent had better activity than the positive control drugs Osocimab and MAA868.

[0236] Table 8 IC value of humanized antibody of antibody 29E10C4 in S2366 detection 50

[0237] The results showed that the IC 50 value of the humanized antibody 29E10C4-1 to be tested was much lower than the positive control, so the humanized antibody to be tested had better activity than the positive control drug MAA868.

[0238] FXIa kit activity detection

[0239] FXIa kit activity detection was performed according to the instructions of FXIa kit (HYPHEN BioMed, BIOPHEN TM Factor Xia). Osocimab and MAA868 antibody were used as positive control, and each antibody to be tested was detected by FXIa kit and IC 50 value was calculated. The results are shown in Figures 6-7 and Tables 9-10. 50

[0240] ​​Table 9 IC of humanized antibody of antibody 14F3 detected by FXIa kit 50

[0241] The experimental results show that the IC of each antibody to be tested 50 is lower than that of the positive control Osocimab and MAA868, especially antibodies 14F3-1, 14F3-Z1 and 14F3-Z2 show IC far lower than the positive control 50 , so the tested antibodies are better than the positive control drugs.

[0242] Table 10 IC of humanized antibody of antibody 29E10C4 detected by FXIa kit 50

[0243] The experimental results show that the IC of each antibody to be tested 50 is far lower than that of the positive control MAA868, so the tested antibodies are better than the positive control drugs in activity.

[0244] APTT prolongation activity detection

[0245] (1) APTT kit detection of humanized antibody of antibody 14F3

[0246] The anticoagulant activity of the antibody to be tested was detected using human plasma (Oscars) and activated partial thromboplastin time (APTT) detection kit (Regen Bio) according to the steps of the kit instructions. MAA868 and Osocimab antibodies were used as positive controls for APTT function detection of humanized antibodies. The antibody to be tested and the positive control drug were diluted with PBS, and the initial concentration was 0.8 mg / mL, and then diluted by four times in turn, and six concentrations were taken for APTT detection.

[0247] The detection results are shown in Figure 8. From the results, it can be seen that with the increase of the concentration of the antibody, the APTT time is increased to different degrees, and the humanized antibody 14F3 can well prolong the clotting time at low concentration compared with the positive control drug, and the positive drug MAA868 is better than the humanized antibody 14F3 at high concentration. The activity of the humanized antibody after modification to prolong the clotting time of human plasma does not decrease significantly, and reaches the level comparable to the positive drug MAA868.

[0248] (2) Prolongation activity detection of humanized antibody of antibody 29E10C4

[0249] The test humanized antibody and positive control were diluted with PBS to a concentration of 80 μg / mL, 40 μg / mL, 20 μg / mL, 15 μg / mL, 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 0, a total of 8 concentrations for APTT detection. Fresh human plasma was used, after adding the corresponding concentration of antibody, the outsourcing company used a fully automatic coagulation analyzer to detect APTT, and the experimental results are shown in Figure 9.

[0250] In summary, in the human plasma APTT detection, the APTT prolonging effect of humanized antibody 29E10C4-1 is stronger than that of positive drug MAA868.

[0251] Biacore affinity assay

[0252] The ability of humanized antibodies to specifically bind to antigens was determined by Biacore, and the KD, Kd, and Ka were calculated by fitting the curve according to the experimental results. The affinities of the positive controls and humanized antibodies for human FXIa factor were detected using Osocimab and MAA868 antibodies as positive controls, and 7 concentrations were diluted 2 times downward based on the maximum antigen concentration, plus 0 concentration, a total of 8 concentrations for biacore affinity assay. The KD, Kd, and Ka results of the antibodies and positive controls in this patent are shown in Table 11:

[0253] Table 11 Biacore results of humanized antibodies

[0254] The experimental results show that the affinity of humanized antibodies for human FXIa factor is stronger than that of positive controls, showing strong affinity.

[0255] Cynomolgus monkey pharmacodynamic experiment of humanized antibodies

[0256] Humanized antibodies 14F3-1 and 29E10C4-1 were selected for administration to cynomolgus monkeys (Junke Zhengyuan). Blood was collected 30 min before administration of the cynomolgus monkeys, and intravenous injection of 3 mg / kg of antibody was administered, followed by collection of plasma at 15 min, 1 h, 3 h, 6 h, 24 h, 48 h, 72 h, 96 h, 7 d, 10 d, and 14 d for subsequent detection. At the same time, fecal occult blood was detected before administration, 4 d, 7 d, and 14 d after administration to determine drug safety.

[0257] After the antibody enters the blood, it inhibits the activity of FXI by binding to the antigen FXI, and the free antigen portion that fails to bind to the antibody still exhibits its original activity. Therefore, by detecting the concentration of free FXI in plasma at each time point after injection of the antibody, the drug efficacy of the antibody in the plasma can be reflected.

[0258] Free FXI detection method is to use the same antibody as the injection to coat the hole plate, then add the blood plasma with multiple dilution, and then use the mouse antibody with different epitope to the used antibody for subsequent ELISA detection. The detection results are shown in Figure 10.

[0259] As shown in the figure, the degree of inhibition of humanized antibody 29E10C4-1 to free FXI is stronger than MAA868 compared with positive drug MAA868.

[0260] Since the maximum safety problem of anticoagulant drugs is bleeding, and after gastrointestinal microbleeding, blood cells can be detected in feces, fecal occult blood detection was performed on feces collected at-3d, 4d, 7d, 10d, 14d before and after injection of cynomolgus monkeys. After detecting the color, properties, fat globules, red blood cells, white blood cells and other indicators of feces, no abnormalities were detected in the fecal occult blood of MAA868 and humanized antibody 29E10C4-1.

[0261] In summary, the activity of humanized antibody 29E10C4-1 is better than positive control MAA868, and it also shows good safety.

[0262] Pharmacokinetics experiment (PK) of humanized antibody in cynomolgus monkeys

[0263] The pharmacokinetic indicators of humanized antibodies 14F3-1 and 29E10C4-1 in cynomolgus monkeys were detected according to the following steps:

[0264] (1) Injection of cynomolgus monkeys (Junkezhengyuan):

[0265] Dosing method: intravenous injection, dosing amount: 3mg / kg;

[0266] (2) Blood sampling points:

[0267] -30min, 15min, 2h, 6h, 24h, 48h, 72h, 96h, 7d, 10d, 14d, 17d, 21d, 24d, 28d, 31d, 35d blood sampling points;

[0268] (3) Blood sampling scheme:

[0269] About 1mL of whole blood sample was collected from the non-dosing limb of the cynomolgus monkey, immediately placed in a vacuum blood collection tube with sodium citrate 9:1 anticoagulant with sample number marked, placed in an ice box for storage, and placed in a 2-8℃ centrifuge within 1h after collection, centrifuged at 3000rpm for 10 minutes, and the supernatant was separated into an EP tube with sample number marked. After sample processing, store in a-60--90℃ refrigerator;

[0270] (4) Treatment of samples: 5 μL of plasma was dissolved in 45 μL of PBS, centrifuged at 200g at 4°C for 10 min, the red blood cells were discarded, and the serum was collected and stored at -80°C for subsequent PK detection;

[0271] (5) Coating antibody: the specific protein F11 was diluted to 1 ug / mL with coating buffer 1xPBS, 100 μL was added to each well (costar 96-well plate), and incubated at 4°C overnight;

[0272] (6) Blocking: the coating solution in the 96-well plate was discarded, and the plate was dried. 200 μL of 2% BSA (diluted with PBST, m / v) was added to each well, and incubated at 37°C for 2h;

[0273] (7) Incubation of primary antibody: the coating solution was discarded, and the plate was dried. 100 μL of the corresponding standard curve sample and serum sample was added to each well, and incubated at 37°C for 1h. The standard curve samples MAA868, 14F3-1 and 29E10C4-1 and the serum samples were diluted with 2% BSA, and the concentration gradient of the standard curve samples MAA868, 14F3-1 and 29E10C4-1 was 0 ng / mL, 1 ng / mL, 2 ng / mL, 4 ng / mL, 8 ng / mL, 16 ng / mL, 32 ng / mL, 64 ng / mL, 128 ng / mL, 256 ng / mL and 512 ng / mL; the serum samples MAA868, 14F3-1 and 29E10C4-1 were diluted by two times at 15 min, 30 min, 1h, 3h, 8h, 24h, 48h, 96h and 168h: 100x and 1000x.

[0274] (8) Washing: the coating solution was removed, and each well was washed with 200 μL of washing buffer PBST (containing 0.05% Tween-20) for 3 times;

[0275] (9) Incubation of secondary antibody: 100 μL of Mouse Anti-Human IgG Fc Antibody [HRP] (Jing Sui) (diluted with 2% BSA 1:10000) was added, and incubated at 37°C for 1h;

[0276] (10) Washing: the secondary antibody was removed, and each well was washed with 200 μL of washing buffer PBST (containing 0.05% Tween-20) for 4 times;

[0277] (11) TMB color development and termination: 100 μL of TMB (Solarbio) was added for color development for 6 min, and 2M sulfuric acid was added to terminate the reaction;

[0278] (13) Reading: the 96-well plate was placed in a microplate reader, and the absorbance value was read at 450 nm;

[0279] (14) Data processing: draw the curve, calculate the antibody concentration at each time point, use DAS2.0 software to calculate the PK parameters.

[0280] The detection results are shown in Figure 11. It can be seen that the half-lives of the humanized antibodies 14F3-1 and 29E10C4-1 are both above 200h, showing similar pharmacokinetics as the control drug MAA868.

[0281] 7.7 Detection of the risk of bleeding of the humanized antibodies in cynomolgus monkeys

[0282] In order to detect the risk of bleeding of the humanized antibodies in cynomolgus monkeys, first, it is detected whether the antibodies 14F3-1 and 29E10C4-1 affect exogenous coagulation. Fresh plasma is collected according to the method in 7.5, and is handed over to a third party to detect the exogenous coagulation index PT (plasma thrombin time) according to the conventional method known to those skilled in the art.

[0283] The experimental results are shown in Figure 12. It can be seen that in the PT (plasma thrombin time) detection after administration in cynomolgus monkeys, similar to MAA868, 14F3-1 and 29E10C4-1 do not affect the PT detection value of cynomolgus monkeys, therefore, both antibodies do not affect the exogenous coagulation pathway, and do not increase the risk of bleeding caused by exogenous pathway inhibition.

[0284] In addition, the fecal occult blood of cynomolgus monkeys before administration and on the 4th day, 7th day, 14th day, 21st day, 28th day and 35th day after administration is detected, and the specific detection indexes are: fecal color, fecal shape, fat globules, red blood cells, white blood cells, and parasite eggs and bodies, etc. The results are that the fecal occult blood of MAA868 and the humanized antibodies 14F3-1 and 29E10C4-1 are not detected to be abnormal. Therefore, the risk of gastrointestinal bleeding of the antibodies 14F3-1 and 29E10C4-1 is low.

[0285] Example 2 Determination of the formulation

[0286] 2.1 Preliminary screening of the formulation

[0287] In order to prepare a liquid formulation, the purified monoclonal antibody 29E10C4-1 is subjected to buffer exchange with a diafiltration buffer containing the expected buffer composition, and in the case required, concentrated to an antibody concentration greater than the target concentration. After reaching the target concentration, excipients are optionally added to the antibody solution as stock solutions. Finally, the protein concentration is adjusted to the target concentration with the final configuration buffer. The specific components of the different liquid pharmaceutical formulations prepared by the present application are shown in Table 12.

[0288] “-” means none.

[0289] “PS20” means polysorbate 20.

[0290] Table 12. Ingredients of different liquid pharmaceutical formulation formulations

[0291] The liquid pharmaceutical formulation formulations in Table 12 were left still at 40°C for 2 weeks, and the physicochemical stability of the antibody in each formulation was determined. The resulting formulations were analyzed for formulation stability by the following methods, and the results are shown in Table 13.

[0292] (1) Turbidity determination: The absorbance (OD405) of 100 μΐ of the sample was determined at 405 nm using an enzyme marker.

[0293] (2) SEC-HPLC was used to detect soluble high molecular weight species (aggregates, HMW) and low molecular weight hydrolysis products (LMW) in the composition. The SEC-HPLC refers to size exclusion chromatography for quantifying protein aggregates and fragments. Monomers, aggregates and fragments are eluted from the column at different times, equipped with a standard UV detector for detection. In the present application, a Shimadzu / Waters LC-20AT / Waters e2695 high performance liquid chromatograph, and an Agilent AdvanceBio SEC300 2.7 μιη chromatographic column was used to implement the method. For example, about 150 mM sodium phosphate solution (15% acetonitrile, for example, about pH = 6.8) was used as the mobile phase to separate the intact monomers, aggregates and hydrolysis products by isocratic elution, and detection was performed at a wavelength of 280 nm.

[0294] (3) Osmotic pressure determination: measured using an osmometer (OSMOMAT 3000-D).

[0295] Table 13. Physicochemical properties of different compositions after 2 weeks of acceleration

[0296] As can be seen from the results, the different formulations of the present application exhibited a clear appearance without any visible particles after undergoing 2 weeks of acceleration experiments; from the SEC results, the proportion of the main protein was close to 95% or more than 95%, or even more than 97%, and the change rate of the main protein was less than 3.5%, or even less than 1%; the proportion of aggregates or fragments was less than 5%, or less than 2%, and the growth rate of aggregates or fragments was less than 2%. Thus, the liquid pharmaceutical formulation formulations of the present application all exhibited good physicochemical stability.

[0297] 2.2 Effects of sorbitol and arginine hydrochloride on the stability of different concentration formulation formulations

[0298] Formulation 1: 150 mg / ml 29E10C4-1 antibody, 25 mM acetate buffer (acetic acid / sodium acetate buffer), pH 5.5, 0.01 mg / ml PS20 and 180 mM sorbitol;

[0299] Formulation 2: 150 mg / ml 29E10C4-1 antibody, 25 mM acetate buffer (acetic acid / sodium acetate buffer), pH 5.5, 0.01 mg / ml PS20 and 110 mM arginine hydrochloride;

[0300] Formulation 1: 150 mg / ml 29E10C4-1 antibody, 25 mM acetate buffer (acetic acid / sodium acetate buffer), pH 5.5, 0.01 mg / ml PS20 and 180 mM sorbitol;

[0301] Formulation 2: 150 mg / ml 29E10C4-1 antibody, 25 mM acetate buffer (acetic acid / sodium acetate buffer), pH 5.5, 0.01 mg / ml PS20 and 110 mM arginine hydrochloride;

[0302] The osmotic pressure, viscosity (viscosity was detected using RheoSense microVISC) and SEC-HPLC data at day 0, day 7, day 14 of each of the above formulations were detected. Table 14 shows the viscosity of Formulation 1 and Formulation 2, and Table 15 shows the stability data of the two formulations at two concentrations after 2 weeks of standing at 40°C.

[0303] Table 14 Viscosity of Formulation 1 and Formulation 2

[0304] Table 15 Stability data of the two formulations at two antibody concentrations after 2 weeks of standing at 40°C

[0305] As can be seen from Table 15, the osmotic pressure of Formulation 1 and Formulation 2 changes little when the concentration is increased, and still has good stability at a concentration of 150 mg / mL.

[0306] 2.3 Effect of sorbitol and arginine hydrochloride species and concentration on the stability of the formulation

[0307] In addition, Formulation 3 and Formulation 4 were configured to further explore the effect of sorbitol and arginine hydrochloride species and concentration on the stability of the formulation. Formulation 1-4 were placed at 40°C for 28 days, and the accelerated stability data at day 0, day 7, day 14, day 21, day 28 and day 42 were measured, as shown in Table 16 below.

[0308] Formulation 3: 75 mg / ml 29E10C4 antibody, 25 mM acetate buffer (acetic acid / sodium acetate buffer), pH = 5.5, 0.01 mg / ml PS20 and 220 mM sorbitol;

[0309] Formulation 4: 75 mg / ml 29E10C4 antibody, 25 mM acetate buffer (acetic acid / sodium acetate buffer), pH = 5.5, 0.01 mg / ml PS20 and 140 mM arginine hydrochloride.

[0310] Table 16 Accelerated stability data of formulations 1-4

[0311] From the above, it can be seen that the liquid pharmaceutical formulations of the present application all exhibit good physicochemical stability.

[0312] The present application has been described by the above examples, but it should be understood that the above examples are only for the purpose of illustration and description, and are not intended to limit the present application to the scope of the described examples. Furthermore, those skilled in the art can understand that the present application is not limited to the above examples, and that more variations and modifications can be made according to the teachings of the present application, which all fall within the scope of the present application claimed. The scope of protection of the present application is defined by the attached claims and their equivalent scope.

Claims

1. A liquid pharmaceutical preparation comprising: Anti-FXI / FXIa antibody or its antigen-binding fragment and buffer, wherein; The anti-FXI / FXIa antibody or its antigen-binding fragment comprises a heavy chain variable region and / or a light chain variable region, wherein, The heavy chain variable region includes: (i) the three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within a peptide whose amino acid sequence is SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, or 83; or (ii) a sequence containing at least one and no more than five amino acid mutations (preferably amino acid substitutions, preferably conserved substitutions) in all three heavy chain complementarity-determining regions (HCDRs) as described in (i); and / or The light chain variable region includes: (i) The three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3 contained within a peptide whose amino acid sequence is SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, or 84; or (ii) The sequence of the three light chain complementarity-determining regions (LCDRs) described in (i) contains at least one and no more than five amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) in the three light chain complementarity-determining regions (LCDRs).

2. The liquid pharmaceutical formulation of claim 1, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment comprises: The peptide containing the three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 within the amino acid sequence SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, or 83; and / or The peptide containing the amino acid sequence SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82 or 84 contains three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2 and LCDR3.

3. A liquid pharmaceutical formulation comprising an anti-FXI / FXIa antibody or its antigen-binding fragment thereof and a buffer solution, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment comprises: a) Heavy chain complementarity-determining region HCDR3, wherein HCDR3 comprises any amino acid sequence from the group consisting of the following amino acid sequences or is composed of any amino acid sequence from the group consisting of the following amino acid sequences: SEQ ID NO: 5, 11, 17, 23, 29, 35, 41, and 47, or wherein HCDR3 comprises an amino acid sequence having one, two, or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO: 5, 11, 17, 23, 29, 35, 41, or 47; and b) Light chain complementarity-determining region LCDR3, wherein the LCDR3 comprises any amino acid sequence from the group consisting of the following amino acid sequences or is composed of any amino acid sequence from the group consisting of the following amino acid sequences: SEQ ID NO:8, 14, 20, 26, 32, 38, 44 and 50, or the LCDR3 comprises an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown as SEQ ID NO:8, 14, 20, 26, 32, 38, 44 or 50.

4. The liquid pharmaceutical formulation of claim 3, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment further comprises: c) Heavy chain complementarity-determining region HCDR1, wherein HCDR1 comprises any amino acid sequence from the group consisting of the following amino acid sequences or any amino acid sequence from the group consisting of the following amino acid sequences: SEQ ID NO:3, 9, 15, 21, 27, 33, 39 and 45, or wherein HCDR1 comprises an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequences shown as SEQ ID NO:3, 9, 15, 21, 27, 33, 39 or 45; d) Heavy chain complementarity-determining region HCDR2, wherein HCDR2 comprises any amino acid sequence from the group consisting of the following amino acid sequences or any amino acid sequence from the group consisting of the following amino acid sequences: SEQ ID NO:4, 10, 16, 22, 28, 34, 40 and 46, or wherein HCDR2 comprises an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequences shown as SEQ ID NO:4, 10, 16, 22, 28, 34, 40 or 46; e) The light chain complementarity-determining region (LCDR1) comprises any amino acid sequence from the group consisting of SEQ ID NO: 6, 12, 18, 24, 30, 36, 42, and 48, or the LCDR1 comprises an amino acid sequence having one, two, or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO: 6, 12, 18, 24, 30, 36, 42, or 48; and f) Light chain complementarity-determining region (LCDR2) functional region, wherein the LCDR2 comprises any amino acid sequence from the group consisting of the following amino acid sequences or any amino acid sequence from the group consisting of the following amino acid sequences: SEQ ID NO: 7, 13, 19, 25, 31, 37, 43 and 49, or the LCDR2 comprises an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequences shown as SEQ ID NO: 7, 13, 19, 25, 31, 37, 43 or 49.

5. A liquid pharmaceutical formulation comprising an anti-FXI / FXIa antibody or its antigen-binding fragment thereof and a buffer, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3; and three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein: HCDR1 contains or is composed of the amino acid sequence shown in SEQ ID NO:3, 9, 15, 21, 27, 33, 39 or 45, or the HCDR1 contains an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO:3, 9, 15, 21, 27, 33, 39 or 45. HCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO:4, 10, 16, 22, 28, 34, 40 or 46, or the HCDR2 comprises an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO:4, 10, 16, 22, 28, 34, 40 or 46. HCDR3 comprises or is composed of the amino acid sequence shown in SEQ ID NO:5, 11, 17, 23, 29, 35, 41 or 47, or the HCDR3 comprises an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO:5, 11, 17, 23, 29, 35, 41 or 47. LCDR1 contains or is composed of an amino acid sequence as shown in SEQ ID NO:6, 12, 18, 24, 30, 36, 42 or 48, or the LCDR1 contains an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO:6, 12, 18, 24, 30, 36, 42 or 48. LCDR2 comprises or is composed of the amino acid sequence shown in SEQ ID NO:7, 13, 19, 25, 31, 37, 43 or 49, or the LCDR2 comprises an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO:7, 13, 19, 25, 31, 37, 43 or 49; and LCDR3 contains or is composed of an amino acid sequence as shown in SEQ ID NO:8, 14, 20, 26, 32, 38, 44 or 50, or the LCDR3 contains an amino acid sequence having one, two or three amino acid mutations (preferably amino acid substitutions, preferably conservative substitutions) compared to the amino acid sequence shown in SEQ ID NO:8, 14, 20, 26, 32, 38, 44 or 50.

6. The liquid pharmaceutical formulation of claim 1 or 2, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment comprises: (1) The amino acid sequence is as shown in SEQ ID NO:51, containing three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3. The peptide with the amino acid sequence shown in SEQ ID NO:52 contains three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2 and LCDR3; (2) The three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within the peptide segments with amino acid sequences as shown in SEQ ID NO: 53, 67, 69, 71, 73, 75, 77, 79, or 81. The three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2 and LCDR3 contained in the peptide segments with amino acid sequences as shown in SEQ ID NO:54,68,70,72,74,76,78,80 or82; (3) The amino acid sequence is as shown in SEQ ID NO:55, containing three heavy chain complementarity-determining regions (HCDRs): HCDR1, HCDR2, and HCDR3. The peptide with the amino acid sequence shown in SEQ ID NO:56 contains three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2 and LCDR3; (4) The three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 contained within the peptide segment whose amino acid sequence is shown in SEQ ID NO:57 or 83. The peptide containing the three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2 and LCDR3, as shown in the amino acid sequence of SEQ ID NO:58 or 84; (5) The amino acid sequence is as shown in SEQ ID NO:59, containing three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3. The peptide with the amino acid sequence shown in SEQ ID NO:60 contains three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2 and LCDR3; (6) The amino acid sequence is as shown in SEQ ID NO:61, containing three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3. The peptide with the amino acid sequence shown in SEQ ID NO:62 contains three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2 and LCDR3; (7) The amino acid sequence is as shown in SEQ ID NO:63, containing three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3. The amino acid sequence of the peptide, as shown in SEQ ID NO:64, contains three light chain complementarity-determining regions (LCDRs): LCDR1, LCDR2, and LCDR3; or (8) The amino acid sequence is as shown in SEQ ID NO:65, containing three heavy chain complementarity-determining regions (HCDRs) HCDR1, HCDR2, and HCDR3. The peptide with the amino acid sequence shown in SEQ ID NO:66 contains three light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3.

7. The liquid pharmaceutical formulation according to any one of claims 1-6, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment comprises: (1) HCDR1 containing SEQ ID NO:3 or consisting thereof, HCDR2, which contains SEQ ID NO:4 or is composed of therein, Contains SEQ ID NO:5 or HCDR3 composed of it, LCDR1 containing SEQ ID NO:6 or composed thereof, LCDR2, which contains SEQ ID NO:7 or is composed of therein, and LCDR3, which contains SEQ ID NO:8 or is composed of therein; (2) HCDR1 containing SEQ ID NO:9 or consisting thereof, HCDR2, which contains SEQ ID NO:10 or consists of it, HCDR3, which contains SEQ ID NO:11 or is composed of therein, LCDR1 containing SEQ ID NO:12 or composed thereof, LCDR2, which contains SEQ ID NO:13 or is composed of therein, and LCDR3 containing SEQ ID NO:14 or composed thereof; (3) HCDR1 containing or consisting of SEQ ID NO:15, HCDR2, which contains SEQ ID NO:16 or is composed of therein, HCDR3, which contains SEQ ID NO:17 or is composed of therein, LCDR1, which contains SEQ ID NO:18 or is composed of therein, LCDR2, which contains SEQ ID NO:19 or is composed of therein, and LCDR3, which contains SEQ ID NO:20 or is composed of therein; (4) HCDR1 containing or consisting of SEQ ID NO:21, HCDR2, which contains SEQ ID NO:22 or is composed of therein, HCDR3, which contains SEQ ID NO:23 or is composed of therein, LCDR1 containing SEQ ID NO:24 or composed thereof, LCDR2, which contains SEQ ID NO:25 or is composed of therein, and LCDR3, which contains SEQ ID NO:26 or is composed of therein; (5) HCDR1 containing or consisting of SEQ ID NO:27, HCDR2, which contains SEQ ID NO:28 or is composed of therein, Contains SEQ ID NO:29 or HCDR3 composed of it, LCDR1 containing SEQ ID NO:30 or composed thereof, LCDR2, which contains SEQ ID NO:31 or is composed of therein, and LCDR3, which contains SEQ ID NO:32 or is composed of therein; (6) HCDR1 containing or consisting of SEQ ID NO:33, HCDR2, which contains SEQ ID NO:34 or is composed of therein, Contains SEQ ID NO:35 or HCDR3 composed of therein, LCDR1 containing SEQ ID NO:36 or composed thereof, LCDR2, which contains SEQ ID NO:37 or is composed of therein, and LCDR3, which contains SEQ ID NO:38 or is composed of therein; (7) HCDR1 containing or consisting of SEQ ID NO:39, HCDR2, which contains SEQ ID NO:40 or is composed of therein, HCDR3, which contains SEQ ID NO:41 or is composed of therein, LCDR1 containing SEQ ID NO:42 or composed thereof, LCDR2, which contains SEQ ID NO:43 or is composed of therein, and LCDR3 containing SEQ ID NO:44 or composed thereof; or (8) HCDR1 containing or consisting of SEQ ID NO:45, HCDR2, which contains SEQ ID NO:46 or is composed of therein, Contains SEQ ID NO:47 or HCDR3 composed of it, LCDR1 containing SEQ ID NO:48 or composed thereof, LCDR2, which contains SEQ ID NO:49 or is composed of therein, and LCDR3, which contains SEQ ID NO:50 or is composed of it.

8. A liquid pharmaceutical formulation comprising an anti-FXI / FXIa antibody or an antigen-binding fragment thereof and a buffer, wherein the anti-FXI / FXIa antibody or the antigen-binding fragment thereof comprises: The heavy chain variable region has at least 80% identity with any amino acid sequence selected from the group consisting of the following amino acid sequences: SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, and 83; preferably, the heavy chain variable region is selected from amino acid sequences that have one or more (preferably no more than 10, more preferably no more than 5) amino acid mutations (preferably amino acid substitutions, more preferably conserved amino acid substitutions) compared to SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, and 83; preferably, the amino acid mutations do not occur in the heavy chain complementarity-determining region; preferably, the heavy chain variable region has the same identity as SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, and 83. CDRs regions with the same amino acid sequence as SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81 or 83, and the non-CDR regions of the heavy chain variable region have at least 80% identity with the non-CDR regions of sequence SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81 or 83; preferably, the heavy chain variable region has any amino acid sequence selected from the group consisting of the following amino acid sequences: SEQ ID NO: 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81 and 83; and / or The light chain variable region has at least 80% identity with any amino acid sequence selected from the group consisting of the following amino acid sequences: SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, and 84; preferably, the light chain variable region is selected from amino acid sequences that have one or more (preferably no more than 10, more preferably no more than 5) amino acid mutations (preferably amino acid substitutions, more preferably conserved amino acid substitutions) compared to SEQ ID NO: 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, or 84; preferably, the amino acid mutations do not occur in the light chain complementarity-determining region; preferably, the light chain variable region has an identity with the amino acid sequence .... The CDRs region of SEQ ID NO:52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, or 84, and the non-CDRs region of the light chain variable region has at least 80% identity with the non-CDRs region of sequence SEQ ID NO:52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, or 84; preferably, the light chain variable region has any amino acid sequence selected from the group consisting of the following amino acid sequences: SEQ ID NO:52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, and 84.

9. The liquid pharmaceutical formulation of claim 8, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein the amino acid sequence pairs of the heavy chain variable region and the light chain variable region are selected from any one of the following groups of amino acid sequence pairs: SEQ ID NO: 51 and 52, SEQ ID NO: 53 and 54, SEQ ID NO: 55 and 56, SEQ ID NO: 57 and 58, SEQ ID NO: 59 and 60, SEQ ID NO: 61 and 62, SEQ ID NO: 63 and 64, SEQ ID NO: 65 and 66, SEQ ID NO: 67 and 68, SEQ ID NO: 69 and 70, SEQ ID NO: 71 and 72, SEQ ID NO: 73 and 74, SEQ ID NO: 75 and 76, SEQ ID NO: 77 and 78, SEQ ID NO: 79 and 80, SEQ ID NO: 81 and 82 and SEQ ID NO: 83 and 84; Preferably, the amino acid sequence pairs of the heavy chain variable region and the light chain variable region are SEQ ID NO:83 and 84.

10. The liquid pharmaceutical formulation according to any one of claims 1-9, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment further comprises a constant region derived from an IgG antibody, an IgM antibody, an IgA antibody, an IgD antibody, or an IgE antibody; preferably, the constant region is derived from an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody.

11. The liquid pharmaceutical formulation according to any one of claims 1-10, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment further comprises a heavy chain constant region, the heavy chain constant region comprising: SEQ ID NO: 85 or an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the sequence SEQ ID NO: 85; or the heavy chain constant region is composed of SEQ ID NO: 85, or is composed of an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the sequence SEQ ID NO: 85; and / or The light chain constant region comprises: SEQ ID NO:86, or an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the sequence SEQ ID NO:86; or the light chain constant region is composed of SEQ ID NO:86, or is composed of an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the sequence SEQ ID NO:

86.

12. The liquid pharmaceutical formulation according to any one of claims 1-11, wherein the anti-FXI / FXIa antibody or its antigen-binding fragment is a murine antibody, a chimeric antibody, a humanized antibody, or a human antibody; and / or The antibody or its antigen-binding fragment is selected from ScFv, Fab, Fab', (Fab')2, Fv fragment, disulfide-linked Fv (dsFv), and diabody.

13. The liquid pharmaceutical preparation according to any one of claims 1-12, wherein, The buffer solution is selected from one or more of acetic acid, acetate, histidine, histidine hydrochloride, citric acid, citrate, phosphoric acid, phosphate, and tris(hydroxymethyl)aminomethane (TRIS); preferably, the buffer solution is selected from one or more of acetic acid, sodium acetate, histidine, sodium histidine, citric acid, sodium citrate, phosphoric acid, disodium hydrogen phosphate, sodium dihydrogen phosphate, and tris(hydroxymethyl)aminomethane; preferably, the buffer solution is selected from one or more of acetic acid, sodium acetate, citric acid, sodium citrate, phosphoric acid, and disodium hydrogen phosphate; preferably, the buffer solution is selected from one or more of acetic acid, sodium acetate, citric acid, and disodium hydrogen phosphate; more preferably, the buffer solution is acetic acid-sodium acetate or citrate-disodium hydrogen phosphate; more preferably, it is acetic acid-sodium acetate.

14. The liquid pharmaceutical formulation according to any one of claims 1-13, further comprising a surfactant and / or a stabilizer, wherein... The surfactant is selected from one or more of poloxamer 188, polysorbate 80, and polysorbate 20; preferably polysorbate 20 or polysorbate 80; preferably polysorbate 20; and / or The stabilizer is selected from one or more of sucrose, trehalose, mannitol, sorbitol, maltose, arginine, and arginine hydrochloride; preferably, the stabilizer is selected from one or more of sucrose, trehalose, mannitol, sorbitol, maltose, and arginine hydrochloride, and more preferably, sorbitol or arginine hydrochloride.

15. The liquid pharmaceutical preparation according to any one of claims 1-14, wherein the pH value is about 5.0 to about 8.0, preferably about 5.0 to about 6.0, and more preferably about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.

0.

16. The liquid pharmaceutical preparation according to any one of claims 1-15, wherein, The concentration of the anti-FXI / FXIa antibody or its antigen-binding fragment is about 70 mg / mL to about 500 mg / mL, preferably about 75 mg / mL to about 500 mg / mL, preferably about 80 mg / mL to about 500 mg / mL, preferably about 100 mg / mL to about 400 mg / mL, preferably about 100 mg / mL, 110 mg / mL, 120 mg / mL, 130 mg / mL, 140 mg / mL, about 150 mg / mL, about 160 mg / mL, about 170 mg / mL, about 180 mg / mL, about 190 mg / mL, about 200 mg / mL, about 210 mg / mL, about 220 mg / mL, about 230 mg / mL, about 240 mg / mL, about 250 mg / mL, about 300 mg / mL, about 350 mg / mL, or about 400 mg / mL; and / or The concentration of the buffer solution is about 1 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM; and / or The concentration of the stabilizer is about 100 mM to about 300 mM, preferably about 100 mM to about 220 mM, more preferably about 110 mM to about 220 mM, more preferably about 120 mM to about 200 mM, and even more preferably about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 210 mM, or about 220 mM; and / or The concentration of the surfactant is from about 0.005 mg / mL to about 0.1 mg / mL, preferably about 0.005 mg / mL, about 0.006 mg / mL, about 0.007 mg / mL, about 0.008 mg / mL, about 0.009 mg / mL, about 0.01 mg / mL, about 0.02 mg / mL, about 0.03 mg / mL, about 0.04 mg / mL, about 0.05 mg / mL, about 0.06 mg / mL, about 0.07 mg / mL, about 0.08 mg / mL, about 0.09 mg / mL, or about 0.1 mg / mL.

17. The liquid pharmaceutical preparation according to any one of claims 1-15, comprising: An anti-FXI / FXIa antibody or its antigen-binding fragment at a concentration of approximately 70 mg / mL to approximately 500 mg / mL, preferably approximately 75 mg / mL to approximately 500 mg / mL, preferably approximately 80 mg / mL to approximately 500 mg / mL, preferably approximately 100 mg / mL to approximately 400 mg / mL, preferably approximately 70 mg / mL to approximately 200 mg / mL, preferably approximately 75 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, 110 mg / mL, 120 mg / mL, 130 mg / mL, 140 mg / mL, approximately 150 mg / mL, approximately 160 mg / mL, approximately 170 mg / mL, approximately 180 mg / mL, approximately 190 mg / mL, or approximately 200 mg / mL; wherein, The anti-FXI / FXIa antibody or its antigen-binding fragment comprises: HCDR1 comprising SEQ ID NO:21 or thereof, HCDR2 comprising SEQ ID NO:22 or thereof, HCDR3 comprising SEQ ID NO:23 or thereof, LCDR1 comprising SEQ ID NO:24 or thereof, LCDR2 comprising SEQ ID NO:25 or thereof, and LCDR3 comprising SEQ ID NO:26 or thereof; preferably, the anti-FXI / FXIa antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the amino acid sequence of HCDR1 is as shown in SEQ ID NO:21, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:22, the amino acid sequence of HCDR3 is as shown in SEQ ID NO:23, the amino acid sequence of LCDR1 is as shown in SEQ ID NO:24, the amino acid sequence of LCDR2 is as shown in SEQ ID NO:25, and the amino acid sequence of LCDR3 is as shown in SEQ ID NO:

26. As shown in NO:26; preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or its antigen-binding fragment are shown in SEQ ID NO:83 and SEQ ID NO:84, respectively; and An acetate / sodium acetate buffer solution or citrate-disodium hydrogen phosphate at a concentration of about 1 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 10 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM; and Sorbitol or arginine hydrochloride at a concentration of about 100 mM to about 300 mM, preferably about 120 mM to about 220 mM, more preferably about 120 mM to about 200 mM, more preferably about 150 mM to about 220 mM, and even more preferably about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, or about 220 mM; and About 0.005 mg / mL to about 0.1 mg / mL, preferably about 0.005 mg / mL, about 0.006 mg / mL, about 0.007 mg / mL, about 0.008 mg / mL, about 0.009 mg / mL, about 0.01 mg / mL, about 0.02 mg / mL, about 0.03 mg / mL, about 0.04 mg / mL, about 0.05 mg / mL, about 0.06 mg / mL, about 0.07 mg / mL, about 0.08 mg / mL, about 0.09 mg / mL, or about 0.1 mg / mL of polysorbate 20 or polysorbate 80; and The pH value is from about 5.0 to about 8.0, preferably from about 5.0 to about 6.0, and more preferably about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.

0.

18. The liquid pharmaceutical preparation according to any one of claims 1-17, comprising: a) Anti-FXI / FXIa antibody or its antigen-binding fragment at a concentration of 150 mg / ml ± 50 mg / ml or 75 mg / ml ± 20 mg / ml; preferably about 150 mg / ml anti-FXI / FXIa antibody or its antigen-binding fragment; preferably about 75 mg / ml anti-FXI / FXIa antibody or its antigen-binding fragment. b) 25 mM ± 5 mM acetate / sodium acetate buffer; preferably about 25 mM acetate / sodium acetate buffer; c) 150 mM ± 10 mM, 180 mM ± 10 mM, or 220 mM ± 10 mM of sorbitol, preferably about 150 to about 220 mM; preferably about 150 mM, about 180 mM, or about 220 mM of sorbitol; and d) 0.01 mg / ml ± 0.005 mg / ml of polysorbate 20; preferably, about 0.01 mg / ml of polysorbate 20; and The pH is 5.3 ± 0.3; preferably 5.5 ± 0.3; preferably about 5.

5. Preferably, the anti-FXI / FXIa antibody or its antigen-binding fragment comprises: HCDR1 comprising SEQ ID NO:21 or thereof, HCDR2 comprising SEQ ID NO:22 or thereof, HCDR3 comprising SEQ ID NO:23 or thereof, LCDR1 comprising SEQ ID NO:24 or thereof, LCDR2 comprising SEQ ID NO:25 or thereof, and LCDR3 comprising SEQ ID NO:26 or thereof; preferably, the anti-FXI / FXIa antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the amino acid sequence of HCDR1 is as shown in SEQ ID NO:21, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:22, the amino acid sequence of HCDR3 is as shown in SEQ ID NO:23, the amino acid sequence of LCDR1 is as shown in SEQ ID NO:24, the amino acid sequence of LCDR2 is as shown in SEQ ID NO:25, and the amino acid sequence of LCDR3 is as shown in SEQ ID NO:

26. As shown in NO:26; preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or its antigen-binding fragment are shown in SEQ ID NO:83 and SEQ ID NO:84, respectively.

19. The liquid pharmaceutical preparation according to any one of claims 1-17, comprising: a) 150 mg / ml ± 50 mg / ml of anti-FXI / FXIa antibody or its antigen-binding fragment; preferably about 150 mg / ml of anti-FXI / FXIa antibody or its antigen-binding fragment; b) 25 mM ± 5 mM citrate-disodium hydrogen phosphate; preferably about 25 mM citrate-disodium hydrogen phosphate; c) 150 mM ± 10 mM sorbitol, preferably about 150 mM sorbitol; and d) 0.05 mg / ml ± 0.01 mg / ml of polysorbate 20; preferably about 0.05 mg / ml of polysorbate 20; and pH 5.3±0.3; pH 5.5±0.3; preferably pH 5.5; Preferably, the anti-FXI / FXIa antibody or its antigen-binding fragment comprises: HCDR1 comprising SEQ ID NO:21 or thereof, HCDR2 comprising SEQ ID NO:22 or thereof, HCDR3 comprising SEQ ID NO:23 or thereof, LCDR1 comprising SEQ ID NO:24 or thereof, LCDR2 comprising SEQ ID NO:25 or thereof, and LCDR3 comprising SEQ ID NO:26 or thereof; preferably, the anti-FXI / FXIa antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the amino acid sequence of HCDR1 is as shown in SEQ ID NO:21, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:22, the amino acid sequence of HCDR3 is as shown in SEQ ID NO:23, the amino acid sequence of LCDR1 is as shown in SEQ ID NO:24, the amino acid sequence of LCDR2 is as shown in SEQ ID NO:25, and the amino acid sequence of LCDR3 is as shown in SEQ ID NO:

26. As shown in NO:26; preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or its antigen-binding fragment are shown in SEQ ID NO:83 and SEQ ID NO:84, respectively.

20. The liquid pharmaceutical preparation according to any one of claims 1-17, comprising: a) Anti-FXI / FXIa antibody or its antigen-binding fragment at a concentration of 150 mg / ml ± 50 mg / ml or 75 mg / ml ± 20 mg / ml; preferably about 150 mg / ml anti-FXI / FXIa antibody or its antigen-binding fragment; preferably about 75 mg / ml anti-FXI / FXIa antibody or its antigen-binding fragment. b) 25 mM ± 5 mM acetate / sodium acetate buffer; preferably about 25 mM acetate / sodium acetate buffer; c) Arginine hydrochloride of about 100 mM to about 150 mM; preferably arginine hydrochloride of about 110 mM to about 140 mM; preferably arginine hydrochloride of 110 mM ± 10 mM or 140 mM ± 10 mM; preferably arginine hydrochloride of about 110 mM or about 140 mM; and d) 0.01 mg / ml ± 0.005 mg / ml of polysorbate 20 or polysorbate 80; preferably about 0.01 mg / ml of polysorbate 20 or polysorbate 80; and The pH is 5.5 ± 0.3; the preferred pH is approximately 5.

5. Preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or its antigen-binding fragment are shown in SEQ ID NO:83 and SEQ ID NO:84, respectively.

21. Liquid pharmaceutical preparations, comprising a) An anti-FXI / FXIa antibody or its antigen-binding fragment at a concentration of approximately 150 mg / ml, wherein the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or its antigen-binding fragment are shown in SEQ ID NO:83 and SEQ ID NO:84, respectively. b) Approximately 25 mM acetate / sodium acetate buffer; c) Approximately 110 mM of arginine hydrochloride; and d) Approximately 0.01 mg / ml of polysorbate 20 or polysorbate 80; and The pH is approximately 5.

5.

22. Liquid pharmaceutical preparations, comprising a) An anti-FXI / FXIa antibody or its antigen-binding fragment at a concentration of approximately 150 mg / ml, wherein the amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-FXI / FXIa antibody or its antigen-binding fragment are shown in SEQ ID NO:83 and SEQ ID NO:84, respectively. b) Approximately 25 mM acetate / sodium acetate buffer; c) Approximately 180 mM of sorbitol; and d) Approximately 0.01 mg / ml of polysorbate 20 or polysorbate 80; and The pH is approximately 5.

5.

23. The liquid pharmaceutical preparation of any one of claims 1-22, wherein the liquid pharmaceutical preparation is administered to a subject by a mode of administration selected from the following: subcutaneous administration, intravenous administration, inhalation administration, intradermal administration, transdermal administration, intraperitoneal administration, and intramuscular administration.

24. A lyophilized formulation prepared by means of a liquid pharmaceutical formulation as described in any one of claims 1-22.

25. A method for preparing a liquid pharmaceutical formulation according to any one of claims 1-22, comprising mixing the anti-FXI / FXIa antibody or its antigen-binding fragment with one or more pharmaceutically acceptable excipients.

26. Use of the liquid pharmaceutical preparation of any one of claims 1-22 or the lyophilized preparation of claim 24 in the preparation of a medicament for treating coagulation or thromboembolism.

27. The liquid pharmaceutical preparation of any one of claims 1-22 or the lyophilized preparation of claim 24, for the treatment of coagulation or thromboembolism.

28. A method for treating coagulation or thromboembolism, comprising administering to a subject in need a therapeutically effective amount of any one of claims 1-22, or the lyophilized formulation of claim 24.

29. A container comprising a liquid pharmaceutical preparation according to any one of claims 1-22 or a lyophilized preparation according to claim 24, wherein the container is a tube, bottle, vial, syringe, or injection pen.

30. A medicine box comprising a container containing a liquid pharmaceutical preparation according to any one of claims 1-22 or a lyophilized preparation according to claim 24.

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