Methods of administering FVIII-mimicking bispecific antibodies every two weeks

Abstract

The present invention relates generally to the use of bispecific FVIII mimicking antibodies in the treatment of hemophilia, such as hemophilia A with or without inhibitors, in particular methods of treating the disease, such as dosage regimens and compositions used in such methods.

Description

[0001] This application is a divisional application of Chinese invention patent application filed on March 1, 2023, with application number 202380024325.0 and invention title "Method for administering FVIII mimicking bispecific antibodies every two weeks". Technical Field

[0002] This invention relates to a method for administering factor VIII mimic antibodies to hemophilia patients.

[0003] Incorporation of sequence lists This application is filed together with the electronic sequence list, the entire contents of which are incorporated herein by reference. Background Technology

[0004] In patients with coagulation disorders, such as those with hemophilia A and B, various steps of the coagulation cascade become dysfunctional due to, for example, the lack or inadequacy of functional clotting factors. This dysfunction of a part of the coagulation cascade leads to inadequate blood clotting and potentially life-threatening bleeding, or damage to internal organs such as joints.

[0005] Factor VIII (FVIII) deficiency, commonly known as hemophilia A, is a congenital bleeding disorder affecting approximately 420,000 people worldwide, of whom about 105,000 are currently diagnosed.

[0006] Patients with hemophilia A can receive clotting factor replacement therapy, such as exogenous FVIII. Routine treatment consists of replacement therapy, which is provided as a preventative measure against bleeding episodes or as needed. Until recently, prophylactic treatment for patients with severe hemophilia A was intravenous administration of plasma-derived FVIII or recombinant FVIII or its long-acting variants up to three times per week.

[0007] However, these patients are at risk of developing neutralizing antibodies (so-called inhibitors) against these exogenous factors, rendering previously effective therapies ineffective. Hemophilia A patients with inhibitors are a non-limiting example of a partially congenital, partially acquired coagulopathy. Patients who have developed FVIII inhibitors cannot be treated with conventional replacement therapy.

[0008] Exogenous clotting factors can only be administered intravenously, which is quite inconvenient and uncomfortable for patients. For example, infants and young children may require surgical insertion of an intravenous catheter into a chest vein to ensure venous access. This exposes them to a significant risk of bacterial infection.

[0009] The drug emicizumab (HEMLIBRA) ®Also known as ACE910, it has been approved for subcutaneous prophylactic treatment of hemophilia A with or without an inhibitor. Emicizumab is a humanized bispecific anti-FIX(a) / anti-FX(a) monoclonal antibody developed by Chugai Pharmaceuticals / Roche Pharmaceuticals for the treatment of hemophilia A. Emicizumab is engineered to mimic the function of cofactor FVIII (see Sampei et al.: (2013)). PLoS One , 8, e57479 and WO2012 / 067176).

[0010] WO2015 / 194233 and WO2018 / 047813 disclose dosage regimens that are allegedly used for administering emecizumab.

[0011] For example, WO2018 / 021450, WO2020 / 025672 and WO2021 / 152066 also disclose FVIII mimicking anti-FIX(a) and anti-FX(a) bispecific antibodies and their use as procoagulants in the treatment of hemophilia A.

[0012] There is a need in the art for improved methods of administering specific bispecific FVIII mimic antibodies to patients with hemophilia A, with or without inhibitors. Summary of the Invention

[0013] This invention relates to a method of administering a compound that acts as a substitute for FVIII in patients suffering from coagulopathy, particularly those lacking functional coagulation factor VIII (FVIII), such as patients with hemophilia A, including those with hemophilia A having an inhibitor. In particular, it relates to a bispecific antibody capable of treating hemophilia A with or without an inhibitor, wherein the bispecific antibody is capable of binding to FIX (SEQ ID NO:1) or its activated form and FX (SEQ ID NO:2) or its activated form.

[0014] In one aspect, the present invention relates to bispecific antibodies for treating hemophilia A with or without inhibitors, wherein the bispecific antibody comprises An anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and Anti-FX(a) antibodies or antigen-binding fragments thereof capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising heavy and light chains, in The heavy chain of the anti-FIX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:3, 4 and 5, respectively, and The light chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:8, 9 and 10, respectively, and The heavy chain of the anti-FX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:13, 14 and 15, respectively, and The light chain of the anti-FX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:18, 19 and 20, respectively, and The bispecific antibody will be administered subcutaneously to human patients in a composition containing the bispecific antibody. The loading dose includes - Approximately 9 mg of the bispecific antibody was administered to patients weighing 5 kg to <15 kg, or - Approximately 29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or - Approximately 66 mg of the described bispecific antibody, administered to patients weighing 45 kg or more. Maintenance dose includes - Approximately 4 mg of the bispecific antibody, administered every two weeks to patients weighing 5 kg to <15 kg, or - Approximately 9 mg of the bispecific antibody, administered every two weeks to patients weighing 15 kg to <45 kg, or - Approximately 20 mg of the bispecific antibody, administered every two weeks to patients weighing 45 kg or more. The first maintenance dose is administered two weeks after the loading dose; and The steady-state plasma concentration of the bispecific antibody is provided in the range of about 2 µg / mL to about 18 µg / mL, such as 3 to 9 µg / mL, such as 6.5 µg / mL.

[0015] In one embodiment, the bispecific antibody comprises a first heavy chain and a first light chain, a second heavy chain and a second light chain, wherein the first heavy chain comprises SEQ ID NO:7, the first light chain comprises SEQ ID NO:12, the second heavy chain comprises SEQ ID NO:17, and the second light chain comprises SEQ ID NO:22 (mAb1).

[0016] In one embodiment, the present invention relates to a pharmaceutical composition comprising a bispecific antibody as described herein.

[0017] In one embodiment, the bispecific antibody is to be administered subcutaneously to a human patient in a pharmaceutical composition comprising the bispecific antibody as described herein.

[0018] On the one hand, as disclosed herein, the bispecific antibody is administered once every two weeks.

[0019] In a preferred embodiment, as disclosed herein, the antibody is administered at a loading dose followed by a maintenance dose.

[0020] In another aspect, the present invention relates to a kit comprising a composition containing the bispecific antibody (e.g., in an injection device) and instructions for use.

[0021] This application also includes the following implementation schemes: 1. A bispecific antibody for treating hemophilia A with or without an inhibitor, wherein the bispecific antibody comprises An anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and Anti-FX(a) antibodies or antigen-binding fragments thereof capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising heavy and light chains, in The heavy chain of the anti-FIX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:3, 4 and 5, respectively, and The light chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:8, 9 and 10, respectively, and The heavy chain of the anti-FX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:13, 14 and 15, respectively, and The light chain of the anti-FX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:18, 19 and 20, respectively, and The bispecific antibody will be administered subcutaneously to human patients in a composition containing the bispecific antibody. The loading dose includes - Approximately 9 mg of the bispecific antibody was administered to patients weighing 5 kg to <15 kg, or - Approximately 29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or - Approximately 66 mg of the described bispecific antibody, administered to patients weighing 45 kg or more. Maintenance dose includes - Approximately 4 mg of the bispecific antibody, administered every two weeks to patients weighing 5 kg to <15 kg, or - Approximately 9 mg of the bispecific antibody, administered every two weeks to patients weighing 15 kg to <45 kg, or - Approximately 20 mg of the bispecific antibody, administered every two weeks to patients weighing 45 kg or more. The first maintenance dose is administered two weeks after the loading dose; and The steady-state plasma concentration of the bispecific antibody is provided in the range of about 2 µg / mL to about 18 µg / mL, such as 3 to 9 µg / mL, such as 6.5 µg / mL.

[0022] 2. A bispecific antibody for treating hemophilia A with or without an inhibitor, as described in Implementation Scheme 1, wherein... The anti-FIX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:6 and a light chain variable domain represented by SEQ ID NO:11, and wherein The anti-FX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:16 and a light chain variable domain represented by SEQ ID NO:21.

[0023] 3. A bispecific antibody for treating hemophilia A with or without an inhibitor, as described in Implementation Scheme 2, wherein... The heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12. The heavy chain of the anti-FX(a) antibody contains SEQ ID NO:17, and the light chain of the anti-FX(a) antibody contains SEQ ID NO:22.

[0024] 4. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the foregoing embodiments, wherein the bispecific antibody is mAb1.

[0025] 5. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the foregoing embodiments, wherein the loading dose comprises -9 mg of the bispecific antibody, administered to patients weighing 5 kg to <15 kg, or -29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or -66 mg of the bispecific antibody, administered to patients weighing 45 kg or more. Maintenance dose includes -4 mg of the bispecific antibody, administered every two weeks to patients weighing 5 kg to <15 kg, or -9 mg of the bispecific antibody, administered every two weeks to patients weighing 15 kg to <45 kg, or - 20 mg of the bispecific antibody, administered every two weeks to patients weighing 45 kg or more. This provides a steady-state plasma concentration of the bispecific antibody in the range of approximately 3 to 9 µg / mL, such as 6.5 µg / mL.

[0026] 6. A bispecific antibody for treating hemophilia A with or without an inhibitor, wherein the bispecific antibody is mAb1, and mAb1 will be administered subcutaneously to human patients in a composition containing mAb1. The loading dose includes -9 mg mAb1, administered to patients weighing 5 kg to <15 kg, or -29 mg mAb1, administered to patients weighing 15 kg to <45 kg, or -66 mg mAb1, administered to patients weighing 45 kg or more. Maintenance dose includes -4 mg mAb1, administered every two weeks to patients weighing 5 kg to <15 kg, or -9 mg mAb1, administered every two weeks to patients weighing 15 kg to <45 kg, or - 20 mg of mAb1, administered every two weeks to patients weighing 45 kg or more. The first maintenance dose is administered two weeks after the loading dose; and This provides steady-state plasma concentrations of mAb1 in the range of 3 to 9 µg / mL, such as 6.5 µg / mL.

[0027] 7. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the foregoing embodiments, wherein the composition comprises about 150 mM of L-arginine hydrochloride or L-arginine, about 20 mM of L-histidine, and a surfactant, such as polysorbate 20 or polysorbate 80, at a pH of about 6.3.

[0028] 8. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the foregoing embodiments, wherein the composition comprises about 150 mM of L-arginine hydrochloride or L-arginine, about 20 mM of L-histidine, and about 0.02 w / v% of polysorbate 20 or polysorbate 80, at a pH of about 6.3.

[0029] 9. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the foregoing embodiments, wherein the composition comprises about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, and about 0.02 w / v% of polysorbate 20, at a pH of about 6.3.

[0030] 10. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the foregoing embodiments, wherein the composition comprises 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, and 0.02 w / v% of polysorbate 20, at a pH of 6.3.

[0031] 11. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the foregoing embodiments, wherein the composition comprises the bispecific antibody at a concentration of about 2 mg / ml, about 5 mg / ml, about 11.25 mg / ml, about 25 mg / ml, or about 57.5 mg / ml.

[0032] 12. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the foregoing embodiments, wherein the treatment provides an annualized bleeding rate of 0, 1, 2, 3 or 4.

[0033] 13. A pharmaceutical composition comprising a bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02 w / v% of polysorbate 20, and a pH of about 6.3.

[0034] 14. The pharmaceutical composition according to embodiment 13, wherein the composition comprises about 2 mg / mL to about 57.5 mg / mL of bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02 w / v% of polysorbate 20, and a pH of about 6.3.

[0035] 15. A kit comprising a) a pharmaceutical composition comprising a bispecific antibody according to any one of embodiments 1-4; and b) instructions for subcutaneous administration of the pharmaceutical composition once every two weeks for the treatment of hemophilia A with or without an inhibitor according to any of the foregoing embodiments. Attached Figure Description

[0036] Figure 1 SEQ ID NO:3-22 is shown in tabular form.

[0037] Figure 2 The mean curve of mAb1 concentration in patient plasma is shown. Pre-dose measurements below the lower limit of quantitation were set as 0. Due to the logarithmic axis, a concentration of 0 was reported as 1e-2 µg / mL. The vertical line represents the two pharmacokinetic (PK) phases. PK phases were performed on days 56–63 for cohorts 1–3 and 5 (once weekly), while days 56–84 were performed on cohort 4 (once every 4 weeks). Mean + / - SEM.

[0038] Figure 3 This section shows a visual prediction check of the fit between the PK model and the data observed in the FRONTIER1 MAD section. Data points are individual mAb1 plasma concentrations over time. The solid line represents the median of the observed data, and the dashed line represents the median mAb1 plasma concentration predicted by the model relative to time. The dotted lines represent the 5th percentile (bottom) and 95th percentile (top) of the model predictions obtained from 1000 trial simulations using the PK model. The model adequately captures the median trend line and variability of the data across all cohorts.

[0039] Figure 4Peak thrombin levels are shown in patients treated with gradually increasing doses of mAb1 and clinically recommended doses of emecizumab (emi). Plasma samples were collected from patients treated with different doses of mAb1 (multiple escalation dose (MAD) cohorts) who were either starting treatment with emecizumab or receiving emecizumab for established prophylaxis at different time points throughout treatment. Potential FVIII activity was neutralized by adding anti-FVIII antibodies, and thrombin generation was performed in vitro. Solid lines represent in vitro samples of human plasma from healthy subjects, which were made hemophilia A-like with anti-FVIII antibodies and infused with different concentrations of mAb1 or emecizumab. Dashed lines represent the mean mAb1 plasma concentration (C1) for each specified cohort. avg (Calculations are based on the PK stage).

[0040] Figure 5 Typical predictive PK curves for mAb1 in subjects with different body weights are shown, using a single loading dose followed by a maintenance dose, as illustrated in Table 8. PK simulations confirm that all subjects should achieve therapeutic range plasma mAb1 concentrations regardless of body weight, defined by a minimum concentration (2 µg / mL) comparable to that of FRONTIER1 MAD cohort 2 (C2) and a concentration defined by C in MAD cohort 5. max The concentration is defined as a maximum concentration (18 µg / mL). This has been confirmed in QW, Q2W, and QM dosing intervals.

[0041] Sequence Summary SEQ ID NO:1 represents the amino acid sequence of human coagulation factor IX.

[0042] SEQ ID NO:2 represents the amino acid sequence of human coagulation factor X.

[0043] SEQ ID NO:3-22 represents the amino acid sequences belonging to or derived from the bispecific antibody “mAb1” mentioned herein, as shown below: SEQ ID NO: 3, 4 and 5 represent the complementarity-determining regions (CDRs) 1-3 of the heavy chain of the anti-FIX(a) antibody component of mAb1, respectively.

[0044] SEQ ID NO:6 represents the heavy chain variable domain (V) of the anti-FIX(a) antibody component of mAb1. H ).

[0045] SEQ ID NO:7 represents the full-length heavy chain of the anti-FIX(a) antibody component of mAb1.

[0046] SEQ ID NO:8, 9 and 10 represent CDRs 1-3 of the light chain of the anti-FIX(a) antibody component of mAb1, respectively.

[0047] SEQ ID NO:11 represents the light chain variable domain (V) of the anti-FIX(a) antibody component of mAb1. L ).

[0048] SEQ ID NO:12 represents the full-length light chain of the anti-FIX(a) antibody component of mAb1.

[0049] SEQ ID NO: 13, 14 and 15 represent CDR 1-3 of the heavy chain of the anti-FX(a) antibody component of mAb1, respectively.

[0050] SEQ ID NO:16 represents the heavy chain variable domain (V) of the anti-FX(a) antibody component of mAb1. H ).

[0051] SEQ ID NO:17 represents the full-length heavy chain of the anti-FX(a) antibody component of mAb1.

[0052] SEQ ID NO: 18, 19 and 20 represent CDRs 1-3 of the light chain of the anti-FX(a) antibody component of mAb1, respectively.

[0053] SEQ ID NO:21 represents the light chain variable domain (V) of the anti-FX(a) antibody component of mAb1. L ).

[0054] SEQ ID NO:22 represents the full-length light chain of the anti-FX(a) antibody component of mAb1.

[0055] In addition to the accompanying electronic sequence list, Figure 1 SEQ ID NO:3-22 is also presented in tabular form. Detailed Implementation

[0056] This invention relates to methods of administering bispecific antibodies and compositions comprising such bispecific antibodies, which act as a substitute for FVIII in patients with coagulopathy, particularly those lacking functional coagulation factor VIII (FVIII), such as patients with hemophilia A, including those with and without inhibitors of FVIII. Specifically, a dosing regimen of once every two weeks is preferred. To facilitate a better understanding of the invention, some terms are first defined.

[0057] The terms “a” or “an” are intended to mean “one or more (one or more types)”. When the term “comprising” and its variations such as “including” precede a descriptive step or element, it is intended to indicate that adding a further step or element is optional and not excluded.

[0058] This document uses the term "about" to mean approximately, roughly, or around. When the term "about" is used in conjunction with a numerical range, it modifies the range by extending the boundaries above and below the stated value. Typically, the term "about" can modify a value upwards or downwards (higher or lower) by 10% above or below the stated value.

[0059] The term "annualized bleeding rate" (ABR) refers to the number of treated bleeding episodes (including spontaneous and traumatic bleeding) experienced by a patient within a defined period of time, extrapolated to one year. For example, two bleeding episodes within six months would indicate an ABR of four.

[0060] The term antibody includes, but is not limited to, bivalent antibodies, such as bispecific antibodies. A full-length antibody contains at least four polypeptide chains: two heavy chains (HC) and two light chains (LC) linked by disulfide bonds. One class of immunoglobulins of particular pharmaceutical interest is IgG. In humans, based on the sequence of their heavy chain constant regions, the IgG class can be divided into four subclasses: IgG1, IgG2, IgG3, and IgG4 in the preferred embodiment. Based on differences in their sequence composition, the light chains can be divided into two types: κ chains and λ chains. An IgG molecule consists of two heavy chains interconnected by two or more disulfide bonds and two light chains, each connected to the heavy chain by a disulfide bond. The IgG heavy chain may contain a heavy chain variable domain (V). H ) and at most three heavy chains constant (C H Domain: C H 1. C H 2 and C H 3. Light chains may contain light chain variable fields (V). L ) and light chain constant domain (C L V H and V L The region can be further subdivided into highly variable regions known as complementary determinant regions (CDRs) or high-variability regions (HvRs), interspersed with more conservative regions known as framing regions (FRs). V H and V LThe domain typically consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The heavy and light chain variable domains containing the hypervariable region (CDR) constitute the structure capable of interacting with the antigen, while the constant region of the antibody mediates the binding of the immunoglobulin to host tissues or factors, including but not limited to various cells of the immune system (effector cells), Fc receptors, and the first component of the C1 complex of the classical complement system, C1q.

[0061] Antibodies or fragments thereof may be defined according to their complementarity-determining regions (CDRs). The term “complementarity-determining region” or “CDR” as used herein refers to the antibody region containing the amino acid residues involved in antigen binding. A CDR can be identified as the region with the highest variability in the amino acid alignment of the antibody's variable domains. Databases such as the Kabat database can be used for CDR identification; for example, a CDR is defined as containing amino acid residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) of the light chain variable domain and 31-35 (H1), 50-65 (H2), and 95-102 (H3) of the heavy chain variable domain; (Kabat et al. 1991; Sequences of Proteins of Immunological Interest, 5th ed., US Department of Health and Human Services, NIH Publication No. 91-3242). Typically, the amino acid residues in this region are numbered using the method described by Kabat et al. (ibid.). In this paper, phrases such as “Kabat position,” “Kabat residue,” and “according to Kabat” refer to this numbering system used for heavy chain variable domains or light chain variable domains, and unless contradicted by the context, this paper uses the numbering according to Kabat.

[0062] As used in this article, "bispecific antibody" refers to an antibody that can bind to two different antigens or two different epitopes on the same antigen.

[0063] The term "fixed dose" for bispecific antibodies refers to the dose administered to patients whose weight falls within a predetermined range (e.g., 15 kg to <45 kg). Therefore, the fixed dose is not provided in mg / kg, but rather in the absolute amount of the bispecific antibody.

[0064] As used herein, the term "human antibody" is intended to include antibodies having a variable domain in which at least a portion of the frame region and / or at least a portion of the CDR region is derived from a human immunoglobulin sequence. For example, a human antibody may have a variable domain in which both the frame region and the CDR region are derived from a human immunoglobulin sequence. Furthermore, if the antibody contains a constant region, that constant region or a portion thereof is also derived from a human immunoglobulin sequence. Preferably, the human antibody is a monoclonal antibody.

[0065] As used herein, “mAb1” refers to a bispecific antibody comprising an anti-FIX(a) antibody capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and an anti-FX(a) antibody capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising a heavy chain and a light chain, wherein the heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12, the heavy chain of the anti-FX(a) antibody comprises SEQ ID NO:17, and the light chain of the anti-FX(a) antibody comprises SEQ ID NO:22. The bispecific antibody comprises CDR sequences represented by SEQ ID NO:3, 4, 5 and 8, 9, 10 and 13, 14, 15 and 18, 19, 20.

[0066] The term "dosing regimen" or "administration regimen" includes a treatment regimen based on a defined set of doses. For example, in one embodiment, the invention describes a dosing regimen for treating hemophilia A with or without an inhibitor, wherein a bispecific antibody is first administered at a loading dose, followed by a maintenance dose containing the same or a lower amount of the bispecific antibody as the loading dose.

[0067] The term “administration” refers to the application of a substance (such as mAb1) to achieve a therapeutic purpose (e.g., treating hemophilia A with or without inhibitors).

[0068] A “dosage” can be administered by a single dose or by multiple consecutive doses. For example, a 60 mg dose can be administered by a single 60 mg dose or by two consecutive 30 mg doses, while a 120 mg dose can be administered, for example, by three consecutive 40 mg doses. The total number of doses administered consecutively within one hour after the first dose constitutes a single dose (e.g., a loading dose that cannot be easily administered by a single dose).

[0069] The terms “FIX (SEQ ID NO:1) and / or its activated form (FIXa)” may also be referred to as “FIX / FIXa” or simply “FIX(a)”.

[0070] The term “FX (SEQ ID NO:2) and / or its activated form (FXa)” may also be referred to as “FX / FXa” or simply “FX(a)”.

[0071] The term "heavy chain" includes full-length heavy chains. Full-length heavy chains include variable region structural domains V. H and three constant region structural domains C H 1. C H 2 and C H 3. V H The domain is located at the amino terminus of the polypeptide, while C H The domain is located at the carboxyl terminus, where C H 3 is closest to the -COOH end.

[0072] As used in this article, the term "light chain" includes full-length light chains. Full-length light chains include variable region structural domains V. L and constant region structural domain C L The variable domain of the light chain is located at the amino terminus of the polypeptide. Light chains, as described herein, include κ and λ chains.

[0073] The term "kit" refers to a packaged product containing components for administering a bispecific antibody (such as mAb1) to treat a condition, along with instructions for use. The kit preferably comprises a box or container holding the kit's components. The box or container is labeled with a protocol approved by the Food and Drug Administration (or the relevant regulatory authority).

[0074] As used herein, the term "loading dose" refers to the first dose of bispecific antibody administered to a patient at the start of a treatment regimen. Typically, a loading dose aims to achieve treatment-relevant plasma concentrations of the bispecific antibody in the patient within a short period of time.

[0075] The term "loading period" refers to a time during which a patient is treated, including the administration of a bispecific antibody to induce a clinical response. Depending on the required dosing frequency, the "loading period" typically lasts from one week to one month and is triggered by the administration of the first loading dose. The loading period precedes the administration of the first maintenance dose.

[0076] As used in this article, the term "maintenance dose" refers to the dose of bispecific antibody administered to a patient at a point in time following the administration of the loading dose.

[0077] The term "maximum plasma concentration" (C max ( ) refers to the highest concentration of bispecific antibodies observed in the patient's plasma after administration of bispecific antibodies to the patient.

[0078] The term "mean plasma concentration" (or "C") avg "" refers to the average plasma concentration of the bispecific antibody during the dosing interval under steady-state conditions.

[0079] The term "steady-state C of mAb1 plasma concentration" max This refers to a state in which the maximum plasma concentration of mAb1 after administration does not differ between doses. In one embodiment, the steady-state C of mAb1 plasma concentration... max Approximately 18 µg / mL. In another embodiment, the steady-state C of mAb1 plasma concentration... max It is approximately 9 µg / mL.

[0080] The term "steady-state C of mAb1 plasma concentration" min This refers to a state in which the minimum plasma concentration of mAb1 after administration does not differ between doses. In one embodiment, the steady-state C of mAb1 plasma concentration... min Approximately 2 µg / mL. In another embodiment, the steady-state C of mAb1 plasma concentration... min It is approximately 3 µg / mL.

[0081] The term "serum or plasma half-life" refers to the time required for half the amount of a substance administered to a patient to be metabolized or eliminated from the patient's serum or plasma through normal biological processes.

[0082] The term "preventive treatment" refers to the application of a therapy for the treatment of hemophilia A with or without an inhibitor, wherein such treatment is intended to control, manage, prevent, or reduce the occurrence and / or severity of one or more symptoms of hemophilia A with or without an inhibitor, such as bleeding episodes, such as one or more spontaneous bleeding episodes, and / or joint damage.

[0083] The terms "treatment" or "management" refer to reducing the frequency of one or more symptoms of hemophilia A, with or without inhibitors, such as spontaneous or uncontrollable bleeding episodes. However, "treatment" does not necessarily mean a cure.

[0084] The term "T" max "" refers to the time it takes for a substance to reach its maximum concentration in a patient's plasma after it has been administered to the patient.

[0085] This invention relates to a method of administering bispecific antibodies that act as a substitute for FVIII in patients with coagulopathy, particularly those lacking functional coagulation factor VIII (FVIII), such as patients with hemophilia A, including those with and without inhibitors. Specifically, it relates to bispecific antibodies or antigen-binding fragments thereof capable of treating hemophilia A with or without inhibitors, wherein the antibodies are capable of binding to FIX (SEQ ID NO:1) or its activated form and FX (SEQ ID NO:2) or its activated form.

[0086] In one such embodiment, the heavy chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:3, 4 and 5, respectively, and the light chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:8, 9 and 10, respectively, and the heavy chain of the anti-FX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:13, 14 and 15, respectively, and the light chain of the anti-FX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:18, 19 and 20, respectively.

[0087] In one such embodiment, the anti-FIX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:6 and a light chain variable domain represented by SEQ ID NO:11, and the anti-FX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:16 and a light chain variable domain represented by SEQ ID NO:21.

[0088] In a preferred embodiment, the bispecific antibody is an IgG4 isotype.

[0089] In a preferred embodiment, the bispecific antibody is a human antibody.

[0090] In one embodiment, the heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12, and the heavy chain of the anti-FX(a) antibody comprises SEQ ID NO:17, and the light chain of the anti-FX(a) antibody comprises SEQ ID NO:22 (also referred to herein as mAb1).

[0091] The properties of the bispecific antibody have been described in WO2020 / 025672, which is incorporated herein by reference.

[0092] As can be clearly seen from the above, bispecific antibodies can therefore be characterized by their CDR sequence, their variable domain sequence, or their complete heavy and light chain sequences. For the avoidance of doubt, reference to mAb1 requires the presence of the heavy and light chains defined by SEQ ID NOs:7 and 12, and 17 and 22 herein. The terms “mAb1” and “bimAb1” are used interchangeably.

[0093] The method disclosed herein involves administering a subcutaneous injection of a bispecific antibody to a patient every two weeks. Preferably, one or more loading doses are administered prior to the two-week dosing regimen.

[0094] Therefore, in a first aspect of the invention, a method for treating hemophilia A with or without an inhibitor is provided, comprising administering an effective amount of a bispecific antibody to a patient in need, the method comprising... a) Administer at least one loading dose of bispecific antibody to the patient; as well as b) Administer at least one maintenance dose of the bispecific antibody to the patient after the final loading dose.

[0095] On one hand, such a bispecific antibody is mAb1.

[0096] In a preferred embodiment, the loading dose and maintenance dose are selected based on the patient's weight, and in particular the patient's weight range (also referred to herein as the "weight band").

[0097] In this implementation scheme, patients weighing between 5 kg and <15 kg are grouped into one group.

[0098] In another such implementation, patients weighing between 15 kg and <45 kg are grouped into one group.

[0099] In yet another such implementation, patients weighing 45 kg or more are grouped together.

[0100] In one implementation, the loading dose and maintenance dose are fixed doses applicable to patients weighing 5 kg to <15 kg.

[0101] In one implementation, the loading dose and maintenance dose are fixed doses applicable to patients weighing 15 kg to <45 kg.

[0102] In one implementation, the loading dose and maintenance dose are fixed doses applicable to patients weighing 45 kg or more.

[0103] In one implementation, the loading dose and maintenance dose, measured in mg, are the same in terms of the amount of bispecific antibody to be delivered.

[0104] In one implementation, the loading dose and maintenance dose, measured in mg, are not the same in terms of the amount of bispecific antibody to be delivered.

[0105] The administration methods disclosed herein include a dosing regimen every two weeks, which includes a patient-specific weight band and a dosage carefully designed by the inventors to allow for safe and effective treatment.

[0106] Due to the long half-life of mAb1, limited inter-subject variability in plasma concentration, a large therapeutic window, and administration close to the maximal effect (as determined by the inventors), using a simplified weight band for dosing instead of continuous weight-based administration is considered feasible. In addition to providing safe and effective hemostatic coverage, fixed-volume injection based on weight bands is more convenient than dosing per kg of body weight, eliminates the need for dose calculations, and reduces the risk of dosing errors. It aims to simplify administration while still taking into account plasma concentration variations related to body weight and drug product strength (see Example 1, Table 6). This administration method is suitable for injection devices, such as—but not limited to—pre-filled pen syringes for subcutaneous administration of mAb1.

[0107] Dosage regimen range once every two weeks In a preferred embodiment, a loading dose containing about 5 mg to about 15 mg, such as 9 mg of bispecific antibody such as mAb1, is administered to patients weighing 5 kg to <15 kg; a loading dose containing about 25 mg to about 35 mg, such as 29 mg of bispecific antibody such as mAb1, is administered to patients weighing 15 kg to <45 kg; and a loading dose containing about 60 mg to about 70 mg, such as 66 mg of bispecific antibody such as mAb1, is administered to patients weighing 45 kg or more.

[0108] In a preferred embodiment, a maintenance dose containing about 2 mg to about 6 mg, such as 4 mg of bispecific antibody such as mAb1, is administered to patients weighing 5 kg to <15 kg; a maintenance dose containing about 7 mg to about 11 mg, such as 9 mg of bispecific antibody such as mAb1, is administered to patients weighing 15 kg to <45 kg; and a maintenance dose containing about 18 mg to about 22 mg, such as 20 mg of bispecific antibody such as mAb1, is administered to patients weighing 45 kg or more.

[0109] In a preferred embodiment, the first maintenance dose is administered two weeks after the loading dose.

[0110] In a preferred embodiment, maintenance doses are continued every two weeks as long as treatment is necessary.

[0111] In one embodiment, a method for treating hemophilia, such as hemophilia A with or without an inhibitor, is provided, wherein the method comprises administering to the patient a composition comprising a bispecific antibody capable of binding to FIX(a) and FX(a), wherein the administration provides a steady-state plasma concentration of the bispecific antibody in the range of 2 µg / mL to about 18 µg / mL, preferably in the range of about 3 µg / mL to about 9 µg / mL, such as 6 to 7 µg / mL, such as 6.5 to 7 µg / mL.

[0112] In a preferred embodiment, the bispecific antibody comprises an anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and an anti-FX(a) antibody or its antigen-binding fragment capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising a heavy chain and a light chain, wherein the heavy chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:3, 4 and 5, respectively, and the light chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:8, 9 and 10, respectively, and the heavy chain of the anti-FX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:13, 14 and 15, respectively, and the light chain of the anti-FX(a) antibody or its antigen-binding fragment comprises CDR1-3 sequences represented by SEQ ID NO:18, 19 and 20, respectively.

[0113] In a more preferred embodiment, the anti-FIX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:6 and a light chain variable domain represented by SEQ ID NO:11, and the anti-FX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:16 and a light chain variable domain represented by SEQ ID NO:21.

[0114] In the most preferred embodiment, the heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12, and the heavy chain of the anti-FX(a) antibody comprises SEQ ID NO:17, and the light chain of the anti-FX(a) antibody comprises SEQ ID NO:22.

[0115] For mAb1 characterized by its heavy and light chain CDR sequences, heavy and light chain variable domain sequences, or full-length heavy and light chain sequences, the loading dose in a bi-weekly dosing regimen is... In one embodiment, the composition containing the bispecific antibody is administered subcutaneously at a loading dose in a dosing regimen once every two weeks, in patients weighing 5 kg to <15 kg, which may be 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4 or 9.5 mg, preferably 9 mg.

[0116] In one embodiment, the composition containing the bispecific antibody is administered subcutaneously at a loading dose in a dosing regimen once every two weeks, in patients weighing 15 kg to <45 kg, which may be 28.5, 28.6, 28.7, 28.8, 28.9, 29, 29.1, 29.2, 29.3, 29.4 or 29.5 mg, preferably 29 mg.

[0117] In one embodiment, the composition containing the bispecific antibody is administered subcutaneously at a loading dose in a dosing regimen once every two weeks, which may be 65.5, 65.6, 65.7, 65.8, 65.9, 66, 66.1, 66.2, 66.3, 66.4 or 66.5 mg, preferably 66 mg, in patients weighing 45 kg or more.

[0118] For mAb1 characterized by its heavy and light chain CDR sequences, heavy and light chain variable domain sequences, or full-length heavy and light chain sequences, the maintenance dose in a bi-weekly dosing regimen is... In one embodiment, the composition containing the bispecific antibody is administered subcutaneously at a maintenance dose in a dosing regimen once every two weeks, which may be 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4 or 4.5 mg, preferably 4 mg, in patients weighing 5 kg to <15 kg.

[0119] In one embodiment, the composition containing the bispecific antibody is administered subcutaneously at a maintenance dose in a dosing regimen of once every two weeks, which may be 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4 or 9.5 mg, preferably 9 mg, in patients weighing 15 kg to <45 kg.

[0120] In one embodiment, the composition containing the bispecific antibody is administered subcutaneously at a maintenance dose in a dosing regimen that is once every two weeks, and in patients weighing 45 kg or more, the dose may be 19.5, 19.6, 19.7, 19.8, 19.9, 20, 20.1, 20.2, 20.3, 20.4 or 20.5 mg, preferably 20 mg.

[0121] In one implementation, the application method disclosed herein provides 1, 2, 3, 4 or 5, or in the range of 0-3 such as 1-3 or 2-3, or in the range of 1-5 such as 1-2, 1-3, 1-4, 2-3, 2-4, 2-5, 3-4, 3-5 or 4-5 for the ABR.

[0122] In one implementation, the administration method includes administering one, two, or three further loading doses—referred to as “extended loading doses”—to distinguish them from the initial loading dose—if the patient does not achieve an adequate clinical response by the end of the initial loading period. The dose and dosing intervals during the extended loading period are generally the same as those during the initial loading period, but may be altered if the attending healthcare professional has reason to believe that the patient may benefit from changes such as increasing the bispecific antibody dose or more frequent dosing.

[0123] In a preferred embodiment, the first maintenance dose is administered two weeks after the loading dose is given to the patient.

[0124] In one implementation, the first maintenance dose is administered two weeks after the loading dose is given to the patient.

[0125] In one implementation, T 1 / 2 It takes about 30.4 days.

[0126] In one implementation, T max It takes about 9.1 days.

[0127] In a preferred embodiment, the treatment disclosed herein is a preventative treatment.

[0128] In one implementation, the administration method disclosed herein reduces such spontaneous bleeding or bleeding episodes in patients prone to spontaneous bleeding or bleeding episodes.

[0129] pharmaceutical preparations In one aspect of the invention, a pharmaceutical composition suitable for the administration methods disclosed herein is provided.

[0130] Such a pharmaceutical composition contains a bispecific antibody, which is preferably present at a concentration of 1 mg / mL to 100 mg / mL, such as 2 mg / mL to 100 mg / mL, such as 2 mg / mL to 60 mg / mL, and at a pH in the range of 5.5 to 7.5, preferably in the range of 6.0 to 6.5, such as about 6.3.

[0131] In a more preferred embodiment, the concentration of the bispecific antibody is 2 mg / mL, 5 mg / mL, 11.25 mg / mL, 25 mg / mL, or 57.5 mg / mL.

[0132] Such pharmaceutical compositions are suitable for—but not limited to—fixed-dose injection devices, such as those configured to administer 0.8 ml per injection.

[0133] In a preferred embodiment, the pharmaceutical composition is an aqueous formulation. In one embodiment, the bispecific antibody is mAb1.

[0134] Pharmaceutical compositions may further comprise one or more of buffer systems, preservatives, tonifying agents, chelating agents, stabilizers, or surfactants, and various combinations thereof. The use of preservatives, isotonic agents, chelating agents, stabilizers, and surfactants in pharmaceutical compositions is well known to those skilled in the art. See Remington: The Science and Practice of Pharmacy, 19th edition, 1995.

[0135] In one embodiment, the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of a bispecific antibody such as mAb1, L-arginine or L-arginine hydrochloride, L-histidine, and a surfactant, with a pH in the range of 5.5 to 7.0.

[0136] In one embodiment, the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of a bispecific antibody such as mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, polysorbate 20 or polysorbate 80, and a pH in the range of 5.5-7.0.

[0137] In one embodiment, the pharmaceutical composition comprises 2 mg / mL, 5 mg / mL, 11.25 mg / mL, 25 mg / mL or 57.5 mg / mL of bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02% of polysorbate 20, and a pH of about 6.3.

[0138] In a preferred embodiment, the pharmaceutical composition comprises about 2 mg / mL, about 5 mg / mL, about 11.25 mg / mL, about 25 mg / mL or about 57.5 mg / mL of bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02% of polysorbate 20, and a pH of about 6.3.

[0139] In a preferred embodiment, the pharmaceutical composition comprises 2 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02% of polysorbate 20, and pH 6.3.

[0140] In another preferred embodiment, the pharmaceutical composition comprises 5 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02% of polysorbate 20, and pH 6.3.

[0141] In another preferred embodiment, the pharmaceutical composition comprises 11.25 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02% of polysorbate 20, and pH 6.3.

[0142] In another preferred embodiment, the pharmaceutical composition comprises 25 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02% of polysorbate 20, and pH 6.3.

[0143] In yet another preferred embodiment, the pharmaceutical composition comprises 57.5 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02% of polysorbate 20, and a pH of 6.3.

[0144] In one embodiment, the pharmaceutical composition comprises 2 mg / mL, 5 mg / mL, 11.25 mg / mL, 25 mg / mL or 57.5 mg / mL of bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02 w / v% of polysorbate 20, and a pH of about 6.3.

[0145] In a preferred embodiment, the pharmaceutical composition comprises about 2 mg / mL, about 5 mg / mL, about 11.25 mg / mL, about 25 mg / mL or about 57.5 mg / mL of bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02 w / v% of polysorbate 20, and a pH of about 6.3.

[0146] In a preferred embodiment, the pharmaceutical composition comprises 2 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02 w / v% of polysorbate 20, and pH 6.3.

[0147] In another preferred embodiment, the pharmaceutical composition comprises 5 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02 w / v% of polysorbate 20, and pH 6.3.

[0148] In another preferred embodiment, the pharmaceutical composition comprises 11.25 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02 w / v% of polysorbate 20, and pH 6.3.

[0149] In another preferred embodiment, the pharmaceutical composition comprises 25 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02 w / v% of polysorbate 20, and pH 6.3.

[0150] In yet another preferred embodiment, the pharmaceutical composition comprises 57.5 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02 w / v% of polysorbate 20, and pH 6.3.

[0151] In one embodiment, a pharmaceutical composition containing a mAb1 concentration of 2 mg / mL is used to administer a 1.6 mg dose of mAb1.

[0152] In one embodiment, a pharmaceutical composition containing a mAb1 concentration of 5 mg / mL is used to administer a 4 mg dose of mAb1.

[0153] In one embodiment, a pharmaceutical composition containing a mAb1 concentration of 11.25 mg / mL is used to administer a 9 mg dose of mAb1.

[0154] In one embodiment, a pharmaceutical composition containing a mAb1 concentration of 25 mg / mL is used to administer a 20 mg dose of mAb1.

[0155] In one embodiment, a pharmaceutical composition containing a mAb1 concentration of 57.5 mg / mL is used to administer a 46 mg dose of mAb1.

[0156] In some embodiments, the pharmaceutical compositions disclosed herein are intended for use in and / or inclusion in an injection device.

[0157] In a preferred embodiment, the injection device is a fixed-dose device, such as a device configured to deliver a single predetermined dose of a pharmaceutical composition or a device configured to deliver multiple predetermined doses of a pharmaceutical composition; the latter is sometimes referred to as a multi-fixed-dose device or a fixed-dose multiple-shot device. In some embodiments, the injection device is a disposable, pre-filled, multi-dose device. In some embodiments, the injection device is a disposable, pre-filled, single-shot device.

[0158] In one embodiment, the pharmaceutical composition of the invention is administered using an injection device comprising a tube with a needle gauge in the range of 26 to 36.

[0159] In one embodiment, the loading dose and / or maintenance dose may be administered as a single injection, wherein the entire loading dose and / or maintenance dose are administered as a single dose, i.e., the entire dose is administered at once.

[0160] In some embodiments, the loading dose and / or maintenance dose are administered in multiple smaller doses, such as a total of two, three, or four smaller doses constituting the full loading dose or maintenance dose. As a non-limiting example, the loading dose of 80 mg of bispecific antibody may be administered in three smaller doses of 60 mg each. Alternatively, for example, consecutive administration of two 50 mg doses and one 80 mg dose may also be considered.

[0161] The pharmaceutical composition can be administered subcutaneously as a solution of at least 0.05 mL to achieve the desired dose in mg.

[0162] For example, a pharmaceutical composition containing 100 mg / mL mAb1 compound requires a volume of 100 µL to deliver a dose of 10 mg.

[0163] The necessary volume will depend on the concentration of the bispecific antibody in the pharmaceutical composition to be administered, as a lower volume usually results in the need for additional injections, while a larger volume usually results in patient discomfort at the injection site. Typically, the volume administered per injection is 0.08 to 1.5 mL, preferably 0.2 to 1 mL, more preferably 0.6 to 0.9 mL, and even more preferably 0.8 mL of injectable solution.

[0164] The pharmaceutical compositions described above can be combined to allow for the administration of a specific dose using a (fixed) injection volume of 0.8 mL per injection—as disclosed herein. For example, two injections of a pharmaceutical composition containing 57.5 mg / mL mAb1 in 0.8 mL doses would allow for a cumulative dose of 92 mg mAb1. In another example, a single injection of a pharmaceutical composition containing 11.25 mg in 0.8 mL doses, followed by a single injection of a pharmaceutical composition containing 57.5 mL doses in 0.8 mL doses, would allow for a cumulative dose of 55 mg mAb1.

[0165] The pharmaceutical composition can be administered at the same or different injection sites.

[0166] In another general aspect, the present invention relates to a kit comprising an injection device that contains a pharmaceutical composition comprising mAb1 and one or more pharmaceutically acceptable carriers. The kit includes instructions for subcutaneous administration of the pharmaceutical composition for the treatment of hemophilia A with or without inhibitors.

[0167] Further implementation plan 1. A dosage regimen for treating hemophilia A with or without an inhibitor, comprising subcutaneous administration of a bispecific antibody at one or more loading doses, the bispecific antibody comprising... An anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and an anti-FX(a) antibody or its antigen-binding fragment capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising a heavy chain and a light chain. The heavy chain of the anti-FIX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:3, 4 and 5, respectively, and The light chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:8, 9 and 10, respectively, and The heavy chain of the anti-FX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:13, 14 and 15, respectively, and The light chain of the anti-FX(a) antibody or its antigen-binding fragment contains CDR1-3 sequences represented by SEQ ID NO:18, 19 and 20, respectively; Then a maintenance dose is administered every two weeks. The loading dose contains the same or higher amount of the bispecific antibody as the maintenance dose.

[0168] 2. The dosage regimen according to embodiment 1, wherein the anti-FIX(a) antibody or its antigen-binding fragment comprises The heavy chain variable domain represented by SEQ ID NO:6 and the light chain variable domain represented by SEQ ID NO:11, and The anti-FX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:16 and a light chain variable domain represented by SEQ ID NO:21.

[0169] 3. The dosage regimen according to implementation scheme 1 or 2, wherein The heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12. The heavy chain of the anti-FX(a) antibody contains SEQ ID NO:17, and the light chain of the anti-FX(a) antibody contains SEQ ID NO:22.

[0170] 4. The dosage regimen according to any one of the foregoing embodiments, wherein the bispecific antibody is administered as a pharmaceutical composition comprising the bispecific antibody.

[0171] 5. The dosage regimen according to any one of the foregoing embodiments, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, L-arginine or L-arginine hydrochloride, L-histidine and a surfactant, at a pH of about 5.5 to about pH 7.

[0172] 6. The dosage regimen according to any one of the foregoing embodiments, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, L-arginine or L-arginine hydrochloride, L-histidine and a surfactant, and a pH of approximately 6.3.

[0173] 7. The dosage regimen according to any one of the foregoing embodiments, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, about 150 mM of L-arginine, about 20 mM of L-histidine, and about 0.02% of polysorbate 20 or 80, with a pH of about 6.3.

[0174] 8. The dosage regimen according to any one of the foregoing embodiments, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine and about 0.02% of polysorbate 20, with a pH of about 6.3.

[0175] 9. The dosage regimen according to any one of the foregoing embodiments, wherein the pharmaceutical composition comprises 2 mg / mL to 60 mg / mL mAb1, such as 2 mg / mL, 5 mg / mL, 11.25 mg / mL, 25 mg / mL or 57.5 mg / mL mAb1, 150 mM L-arginine hydrochloride, 20 mM L-histidine, and 0.02% polysorbate 20, at pH 6.3.

[0176] 10. The dosage regimen according to any one of the foregoing embodiments, wherein the bispecific antibody is administered to a human patient.

[0177] 11. A dosage regimen according to any one of embodiments 1-10, wherein the loading dose comprises about 5 mg to about 15 mg of the bispecific antibody, administered to a patient weighing 5 kg to <15 kg, or The bispecific antibody, approximately 25 mg to approximately 35 mg, is administered to patients weighing 15 kg to <45 kg, or The bispecific antibody, approximately 60 mg to approximately 70 mg, is administered to patients weighing 45 kg or more. Maintenance dose includes The bispecific antibody, approximately 2 mg to approximately 6 mg, is administered every two weeks to patients weighing 5 kg to <15 kg, or The bispecific antibody, approximately 7 mg to approximately 11 mg, is administered every two weeks to patients weighing 15 kg to <45 kg, or The bispecific antibody, approximately 18 mg to approximately 22 mg, is administered every two weeks to patients weighing 45 kg or more. The first maintenance dose is administered two weeks after the loading dose.

[0178] 12. The dosage regimen according to implementation scheme 11, wherein Administer a loading dose of the bispecific antibody of 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, or 9.5 mg to patients weighing 5 kg to <15 kg, or Administer a loading dose of the bispecific antibody of 28.5, 28.6, 28.7, 28.8, 28.9, 29, 29.1, 29.2, 29.3, 29.4, or 29.5 mg to patients weighing 15 kg to <45 kg, or Administer a loading dose of the bispecific antibody of 65.5, 65.6, 65.7, 65.8, 65.9, 66, 66.1, 66.2, 66.3, 66.4, or 66.5 mg to patients weighing 45 kg or more, and in For patients weighing 5 kg to <15 kg, administer a maintenance dose of the bispecific antibody at a dose of 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, or 4.5 mg every two weeks, or For patients weighing 15 kg to <45 kg, administer a maintenance dose of the bispecific antibody in doses of 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, or 9.5 mg every two weeks, or Patients weighing 45 kg or more shall be given a maintenance dose of the bispecific antibody every two weeks in doses of 19.5, 19.6, 19.7, 19.8, 19.9, 20, 20.1, 20.2, 20.3, 20.4 or 20.5 mg.

[0179] 13. According to the dosage regimen described in embodiment 12, a loading dose of approximately 9 mg of the bispecific antibody is administered to a patient weighing 5 kg to <15 kg, or A loading dose of approximately 29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or A loading dose of approximately 66 mg of the bispecific antibody was administered to patients weighing 45 kg or more, and in which Patients weighing 5 kg to <15 kg should receive a maintenance dose of approximately 4 mg of the bispecific antibody every two weeks, or Patients weighing 15 kg to <45 kg should receive a maintenance dose of approximately 9 mg of the bispecific antibody every two weeks, or Patients weighing 45 kg or more should be given a maintenance dose of approximately 20 mg of the bispecific antibody every two weeks.

[0180] 14. The dosage regimen according to any one of the foregoing embodiments, wherein the treatment is a preventive treatment.

[0181] 15. The dosage regimen according to any one of the foregoing embodiments, wherein a steady-state plasma concentration of the bispecific antibody is provided in the range of about 2 µg / mL to about 18 µg / mL.

[0182] 16. The dosage regimen according to any one of the foregoing embodiments, wherein a steady-state plasma concentration of the bispecific antibody is provided in the range of about 3 µg / mL to about 9 µg / mL, such as 6.5 µg / mL to 7 µg / mL.

[0183] 17. The dosage regimen according to any one of the foregoing embodiments, wherein a steady-state plasma concentration of the bispecific antibody of about 6.5 µg / mL is provided.

[0184] 18. The dosing regimen according to any one of the foregoing embodiments, wherein the regimen provides an ABR of 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 or 3 or 4, or an ABR in the range of 1-5.

[0185] 19. The dosage regimen according to any one of the foregoing embodiments, wherein the bispecific antibody is mAb1.

[0186] 20. A kit comprising a) a pharmaceutical composition comprising a bispecific antibody according to any one of embodiments 1-3; and b) instructions for subcutaneous administration of the pharmaceutical composition once every two weeks for the treatment of hemophilia A with or without an inhibitor according to any of the foregoing embodiments.

[0187] 21. A treatment for hemophilia A, with or without an inhibitor, comprising subcutaneous administration of a bispecific antibody in one or more loading doses, the bispecific antibody containing... An anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and an anti-FX(a) antibody or its antigen-binding fragment capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising a heavy chain and a light chain. The heavy chain of the anti-FIX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:3, 4 and 5, respectively, and The light chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:8, 9 and 10, respectively, and The heavy chain of the anti-FX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:13, 14 and 15, respectively, and The light chain of the anti-FX(a) antibody or its antigen-binding fragment contains CDR1-3 sequences represented by SEQ ID NO:18, 19 and 20, respectively; Then a maintenance dose is administered every two weeks. The loading dose contains the same or a higher amount of the bispecific antibody as the maintenance dose, and Plasma concentrations of the bispecific antibody are provided in the range of about 2 µg / mL to about 18 µg / mL.

[0188] 22. The treatment method according to embodiment 21, wherein the anti-FIX(a) antibody or its antigen-binding fragment comprises The heavy chain variable domain represented by SEQ ID NO:6 and the light chain variable domain represented by SEQ ID NO:11, and The anti-FX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:16 and a light chain variable domain represented by SEQ ID NO:21.

[0189] 23. The treatment method according to implementation scheme 21 or 22, wherein The heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12. The heavy chain of the anti-FX(a) antibody contains SEQ ID NO:17, and the light chain of the anti-FX(a) antibody contains SEQ ID NO:22.

[0190] 24. The treatment method according to any one of embodiments 21-23, wherein the bispecific antibody is administered as a pharmaceutical composition comprising the bispecific antibody.

[0191] 25. The treatment method according to any one of embodiments 21-24, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, L-arginine or L-arginine hydrochloride, L-histidine, and a surfactant, at a pH of about 5.5 to about pH 7.

[0192] 26. The treatment method according to any one of embodiments 21-25, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, L-arginine or L-arginine hydrochloride, L-histidine, and a surfactant, at a pH of approximately 6.3.

[0193] 27. The treatment method according to any one of embodiments 21-26, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, about 150 mM of L-arginine, about 20 mM of L-histidine, and about 0.02% of polysorbate 20 or 80, with a pH of about 6.3.

[0194] 28. The treatment method according to any one of embodiments 21-27, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine and about 0.02% of polysorbate 20, at a pH of about 6.3.

[0195] 29. The treatment method according to any one of embodiments 21-28, wherein the pharmaceutical composition comprises 2 mg / mL to 60 mg / mL mAb1, such as 2 mg / mL, 5 mg / mL, 11.25 mg / mL, 25 mg / mL or 57.5 mg / mL mAb1, 150 mM L-arginine hydrochloride, 20 mM L-histidine, and 0.02% polysorbate 20, at pH 6.3.

[0196] 30. The treatment method according to any one of embodiments 21-29, wherein the bispecific antibody is administered to a human patient.

[0197] 31. The treatment method according to any one of embodiments 21-30, wherein the loading dose comprises about 5 mg to about 15 mg of the bispecific antibody, administered to a patient weighing 5 kg to <15 kg, or The bispecific antibody, approximately 25 mg to approximately 35 mg, is administered to patients weighing 15 kg to <45 kg, or The bispecific antibody, approximately 60 mg to approximately 70 mg, is administered to patients weighing 45 kg or more. Maintenance dose includes The bispecific antibody, approximately 2 mg to approximately 6 mg, is administered every two weeks to patients weighing 5 kg to <15 kg, or The bispecific antibody, approximately 7 mg to approximately 11 mg, is administered every two weeks to patients weighing 15 kg to <45 kg, or The bispecific antibody, approximately 18 mg to approximately 22 mg, is administered every two weeks to patients weighing 45 kg or more. The first maintenance dose is administered two weeks after the loading dose.

[0198] 32. The treatment method according to implementation plan 31, wherein Administer a loading dose of the bispecific antibody of 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, or 9.5 mg to patients weighing 5 kg to <15 kg, or Administer a loading dose of the bispecific antibody of 28.5, 28.6, 28.7, 28.8, 28.9, 29, 29.1, 29.2, 29.3, 29.4, or 29.5 mg to patients weighing 15 kg to <45 kg, or Administer a loading dose of the bispecific antibody of 65.5, 65.6, 65.7, 65.8, 65.9, 66, 66.1, 66.2, 66.3, 66.4, or 66.5 mg to patients weighing 45 kg or more, and in For patients weighing 5 kg to <15 kg, administer a maintenance dose of the bispecific antibody at a dose of 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, or 4.5 mg every two weeks, or For patients weighing 15 kg to <45 kg, administer a maintenance dose of the bispecific antibody in doses of 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, or 9.5 mg every two weeks, or Patients weighing 45 kg or more shall be given a maintenance dose of the bispecific antibody every two weeks in doses of 19.5, 19.6, 19.7, 19.8, 19.9, 20, 20.1, 20.2, 20.3, 20.4 or 20.5 mg.

[0199] 33. The treatment method according to embodiment 32, wherein a loading dose of approximately 9 mg of the bispecific antibody is administered to a patient weighing 5 kg to <15 kg, or A loading dose of approximately 29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or A loading dose of approximately 66 mg of the bispecific antibody was administered to patients weighing 45 kg or more, and in Patients weighing 5 kg to <15 kg should receive a maintenance dose of approximately 4 mg of the bispecific antibody every two weeks, or Patients weighing 15 kg to <45 kg should receive a maintenance dose of approximately 9 mg of the bispecific antibody every two weeks, or Patients weighing 45 kg or more should be given a maintenance dose of approximately 20 mg of the bispecific antibody every two weeks.

[0200] 34. The treatment method according to any one of embodiments 21-33, wherein a plasma concentration of mAb1 is provided in the range of about 3 µg / mL to about 9 µg / mL, such as 6.5 µg / mL to 7 µg / mL.

[0201] 35. The treatment method according to any one of embodiments 21-34, wherein a plasma concentration of mAb1 of about 6.5 µg / mL is provided.

[0202] 36. The treatment method according to any one of embodiments 21-35, wherein the plasma concentration is a steady-state plasma concentration.

[0203] 37. The treatment method according to any one of embodiments 21-36, wherein the treatment provides an ABR of 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 or 4, or an ABR in the range of 1-5.

[0204] 38. The treatment method according to any one of embodiments 24-27, wherein the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of the bispecific antibody, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, and about 0.02 w / v% of polysorbate 20, with a pH of about 6.3.

[0205] 39. The treatment method according to any one of embodiments 24-28, wherein the pharmaceutical composition comprises 2 mg / mL to 60 mg / mL mAb1, such as 2 mg / mL, 5 mg / mL, 11.25 mg / mL, 25 mg / mL or 57.5 mg / mL mAb1, 150 mM L-arginine hydrochloride, 20 mM L-histidine, and 0.02 w / v% polysorbate 20 at pH 6.3.

[0206] 40. The treatment method according to any one of embodiments 21-39, wherein the bispecific antibody is mAb1.

[0207] 41. A pharmaceutical composition comprising a bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02 w / v% of polysorbate 20, and a pH of about 6.3.

[0208] 42. The pharmaceutical composition according to embodiment 41, wherein the composition comprises about 2 mg / mL to about 57.5 mg / mL of bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, about 0.02 w / v% of polysorbate 20, and a pH of about 6.3.

[0209] 43. A kit comprising a) a pharmaceutical composition comprising a bispecific antibody according to any one of embodiments 1-19; and b) instructions for subcutaneous administration of the pharmaceutical composition once every two weeks for the treatment of hemophilia A with or without an inhibitor according to any one of embodiments 21-39.

[0210] 44. The kit according to embodiment 20 or 43, wherein the bispecific antibody is mAb1.

[0211] In a preferred embodiment, the inventors provide a bi-weekly dosing regimen for treating hemophilia A with or without an inhibitor, comprising subcutaneous administration of a loading dose of a bispecific antibody containing… An anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and an anti-FX(a) antibody or its antigen-binding fragment capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising a heavy chain and a light chain. The heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12. The heavy chain of the anti-FX(a) antibody contains SEQ ID NO:17, and the light chain of the anti-FX(a) antibody contains SEQ ID NO:22; Then administer a maintenance dose every two weeks; The bispecific antibody is administered as a pharmaceutical composition comprising the bispecific antibody at a concentration of 5 mg / mL, 11.25 mg / mL, or 25 mg / mL, 150 mM L-arginine hydrochloride, 20 mM L-histidine, and 0.02% polysorbate 20, with a pH of approximately 6.3. in A loading dose of approximately 9 mg of the bispecific antibody was administered to patients weighing 5 kg to <15 kg, or A loading dose of approximately 29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or A loading dose of approximately 66 mg of the bispecific antibody was administered to patients weighing 45 kg or more, and in which Patients weighing 5 kg to <15 kg should receive a maintenance dose of approximately 4 mg of the bispecific antibody every two weeks, or Patients weighing 15 kg to <45 kg should receive a maintenance dose of approximately 9 mg of the bispecific antibody every two weeks, or Patients weighing 45 kg or more should receive a maintenance dose of approximately 20 mg of the bispecific antibody every two weeks. This provides a steady-state plasma concentration of the bispecific antibody in the range of about 2 µg / mL to about 18 µg / mL.

[0212] In a preferred embodiment, the inventors provide a bi-weekly dosing regimen for treating hemophilia A with or without an inhibitor, comprising subcutaneous administration of a loading dose of a bispecific antibody containing… An anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and an anti-FX(a) antibody or its antigen-binding fragment capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising a heavy chain and a light chain. The heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:12. The heavy chain of the anti-FX(a) antibody contains SEQ ID NO:17, and the light chain of the anti-FX(a) antibody contains SEQ ID NO:22; Then administer a maintenance dose every two weeks; The bispecific antibody is administered as a pharmaceutical composition comprising the bispecific antibody at a concentration of 5 mg / mL, 11.25 mg / mL, or 25 mg / mL, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, and 0.02 w / v% of polysorbate 20, with a pH of approximately 6.3. in A loading dose of approximately 9 mg of the bispecific antibody was administered to patients weighing 5 kg to <15 kg, or A loading dose of approximately 29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or A loading dose of approximately 66 mg of the bispecific antibody was administered to patients weighing 45 kg or more, and in which Patients weighing 5 kg to <15 kg should receive a maintenance dose of approximately 4 mg of the bispecific antibody every two weeks, or Patients weighing 15 kg to <45 kg should receive a maintenance dose of approximately 9 mg of the bispecific antibody every two weeks, or Patients weighing 45 kg or more should receive a maintenance dose of approximately 20 mg of the bispecific antibody every two weeks. This provides a steady-state plasma concentration of the bispecific antibody in the range of about 2 µg / mL to about 18 µg / mL.

[0213] Those skilled in the art will understand the methods used to measure and calculate the pharmacokinetics described herein (e.g., but not limited to C). max ,、T max Various methods for performing statistical comparisons (e.g., but not limited to comparisons of changes from baseline to post-treatment and / or comparisons between treatment groups) and / or analyzing the pharmacokinetic and pharmacodynamic parameters described herein will be available.

[0214] Example List of abbreviations

[0215] Example 1: Dose escalation study The following anti-FIX(a) / FX(a) bispecific antibody is being developed for patients with hemophilia A (PwHA) with or without an inhibitor, comprising a first heavy chain and a first light chain, and a second heavy chain and a second light chain, wherein the first heavy chain comprises SEQ ID NO:7, the first light chain comprises SEQ ID NO:12, the second heavy chain comprises SEQ ID NO:17, and the second light chain comprises SEQ ID NO:22 (mAb1).

[0216] FRONTIER1 (EudraCT:2019-000465-20; NCT04204408) aims to investigate the safety, tolerability, pharmacokinetics, pharmacodynamics, and efficacy of a single-escalation subcutaneous dose of mAb1 (a bispecific antibody) in healthy participants, as well as multiple-escalation doses of mAb1 in PwHA with or without inhibitors. Additionally, this study aims to provide data for dose setting in subsequent FRONTIER studies.

[0217] Methods: In the single-escalation-dose (SAD) phase, healthy subjects received a single escalation dose of mAb1 (target plasma concentrations of 0.05 to 3 µg / mL) or placebo. In the multiple-escalation-dose (MAD) phase, PwHA received multiple escalation doses of mAb1, targeting mean plasma concentrations of 1 µg / mL (cohort 1, QW), 3 µg / mL (cohort 2, QW), 9 µg / mL (cohort 3, QW, and cohort 4, Q4W), or 16 µg / mL (cohort 5, QW).

[0218] Nonlinear mixed-effects modeling was used to analyze mAb1 plasma concentration-time data from FRONTIER1. The development of the structural basis model for mAb1 included both a one-compartment and a two-compartment model. The mAb1 plasma concentration-time curves were best described using a two-compartment model with first-order elimination and absorption rates, parameterized as: absorption rate constant (…). k a ), whole-body clearance rate ( CL ), inter-room clearance rate ( Q ), central distribution volume ( V 2 ), peripheral distribution volume ( V 3 ) and relative bioavailability ( F (Fixed to 1). Random effects were explored using inter-individual variability (IIV, or between-subjects variability) on the model's key parameters, as well as accumulated and / or proportional residual errors. In addition to model stability and parameter accuracy, a minimum decrease of 3.84 in the objective function value (OFV) was considered to incorporate random effects. The final model includes... CL , k a and F The residual error on the IIV is described by the proportional error.

[0219] Table 1-5 lists the dosing regimens used in MAD cohorts 1-5.

[0220] Table 1: Loading and maintenance doses of MAD cohort 1 (once-weekly dosing) The loading doses administered in week 0 and week 1 will be numbered 1 and 2.

[0221] Based on body weight measured at week 0.

[0222] The maintenance dose will be administered starting from week 2.

[0223] Based on the latest weight measured according to the scheme flowchart.

[0224] Table 2: Loading and maintenance doses in MAD cohort 2 (once-weekly dosing) The loading doses administered in week 0 and week 1 will be numbered 1 and 2:

[0225] Based on body weight measured at week 0.

[0226] The maintenance dose to be administered starting from week 2:

[0227] Table 3: Loading and maintenance doses in MAD cohort 3 (once-weekly dosing) The loading doses administered in week 0 and week 1 will be numbered 1 and 2:

[0228] The maintenance dose to be administered starting from week 2:

[0229] Table 4: MAD Cohort 4 (Q4W Dosing) A loading dose to be administered in week 0:

[0230] Subsequent doses to be administered starting in week 4:

[0231] Table 5: MAD Cohort 5 (QW Dosing) A loading dose to be administered in week 0:

[0232] The maintenance dose to be administered starting from week 2:

[0233] Results: mAb1 was well tolerated after both single and multiple doses, and no associated thromboembolic events or serious adverse events were reported. No anti-mAb1 antibodies were reported. Data from the SAD section are provided, along with estimated T... 1 / 2 It is 30.4 days, while T max The effective period was 9.1 days. The highest mAb1 concentration observed in MAD cohort 5 was 18 µg / mL, which was evaluated as safe and effective. Figure 2 ).

[0234] Population PK model analysis of the FRONTIER1 data suggests that a two-compartment PK model with first-order absorption and elimination is best described for mAb1. The main covariates affecting mAb1 concentration are baseline body weight and drug product strength. Table 6 collects parameter estimates from the population PK model analysis, while... Figure 3 The model is shown to fit the FRONTIER1 MAD data.

[0235] Table 6: Parameter estimates for the group PK model

[0236] RSE: Relative Standard Error During the 12-week observation period, 15 cases of treated bleeding were reported in 8 patients, of which 13 cases were observed in 6 patients from the lowest dose cohort (9 of which were traumatic bleeding). The two bleeding events in patients from cohorts 2 and 3 were both traumatic bleeding, therefore no treated joint bleeding or spontaneous bleeding was observed outside of cohort 1 (Table 7).

[0237] Conclusion: Pharmacokinetic properties support the bi-weekly dosing regimen as disclosed herein. The population PK model adequately described the median trend and variability of the data, with baseline body weight and drug product strength as the primary covariates. mAb1 was well tolerated, and no anti-mAb1 antibodies were reported. FRONTIER1 provides encouraging data supporting further clinical development using the dosing regimens / dosing schemes described herein, which were derived taking into account the effects of body weight and drug product strength.

[0238] Table 7: Bleeding during 12 weeks of treatment with multi-escalation mAb1 for PwHA

[0239] Example 2: Thrombin generation after mAb1 administration Factor VIII (FVI11) replacement therapy is the standard treatment for patients with hemophilia A (HA). mAb1 is a bispecific antibody that binds to both factor IX(a) and FX(a) and exhibits enhanced hemostatic properties in vitro and in HA mouse models compared to emecizumab. FRONTIER1 (NCT04204408) is a phase 1 / 2 study designed to investigate the safety, tolerability, pharmacokinetics, and pharmacodynamics of subcutaneously administered mAb1 in healthy volunteers and patients with severe HA (regardless of FVI11 inhibitor status).

[0240] Peak thrombin generation and laboratory biomarkers in response to mAb1 or emecizumab were analyzed.

[0241] Methods: The Phase II portion of FRONTIER1 was an open-label study involving subcutaneous administration of mAb1 over 12 weeks in five multi-escalation dose cohorts, with the goal of achieving mean plasma concentrations of 1–18 µg / mL via weekly dosing (cohorts 1–3 and 5) or every four weeks (cohort 4); cohorts 3 and 4 aimed for the same plasma concentrations. An additional exploratory cohort of subjects treated with emecizumab was also included for comparison.

[0242] Results: The study included 42 subjects using mAb1 (cohorts 1 [n=7], 2 [n=9], 3 [n=8], 4 [n=8], and 5 [n=10]) and 10 subjects using emecizumab. Peak thrombin levels increased with increasing mAb1 dose. In in vitro experiments with either mAb1 or emecizumab added to plasma samples neutralized with FVIIl before administration, mAb1 achieved maximum peak height at lower plasma concentrations compared to emecizumab, indicating greater potency (see [link to study]). Figure 4 In cohort 2, mean peak thrombin levels were comparable between patients using emecizumab and mAb1. No dose-dependent changes in D-dimer, fibrinogen, platelet, or FIXa / FX antigen levels were observed; most values ​​remained within the normal range. For patients using mAb1 and emecizumab, dose-dependent increases in prothrombin fragments 1 and 2 were observed (relative changes [%]: 26.08, 93.99, 323.65, 606.41, and 315.62 in cohorts 1–5, and 85.04 for emecizumab), stabilizing at steady state.

[0243] Conclusion: A dose-dependent increase in thrombin generation was observed in mAb1 patients, with peak thrombin levels reaching higher levels than in patients receiving emecizumab. Laboratory parameters showed no safety signals.

[0244] Example 3: Extrapolating the results of Examples 1 and 2 to novel dosing regimens for different body weight bands Based on new insights gained from the FRONTIER1 clinical trials, such as those included in Examples 1 and 2, the inventors carefully analyzed and used the new data obtained from those trials, including patient responses and observed properties of the bispecific antibody mAb1 (e.g., dose, T). ½ T max(and plasma concentrations), and designed administration methods as disclosed herein, particularly specific dosing regimens (including the selection of weight bands / groups). For example, the inventors have achieved therapeutic steady-state plasma concentrations of about 2 µg / mL to about 18 µg / mL, preferably in the range of 3 to 9 µg / mL, such as 5, 5.5, 6, 6.5, or 7 µg / mL, and have developed the necessary dosing regimens to achieve these steady-state plasma concentrations for specific weight bands by increasing the number of weight bands from two to three, to cover weights of 5 kg and above, and thus also include pediatric patients.

[0245] Using the population PK model described in Example 1, the inventors have derived a dosing regimen that rapidly establishes steady-state mAb1 plasma concentrations within the therapeutic range (2 to 18 µg / mL) with a loading dose, followed by a maintenance dose at a Q2W dosing frequency for typical body weight within the hemophilia population.

[0246] Examples of the dosing regimens and weight bands are shown in Table 8.

[0247] Table 8: Examples of maintenance and loading doses of mAb1 and weight bands

[0248] Q2W = every two weeks.

[0249] Figure 5 Examples of mAb1 PK curves for typical subjects within each weight band and dosing frequency are shown in the figure (C2 = cohort 2, C...). avg =Mean plasma concentration, C max =Maximum plasma concentration).

[0250] Example 4: Stability of mAb1 Composition The pharmaceutical composition analyzed in this embodiment contains 1-100 mg / mL of bispecific antibody mAb1, 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, 0.02 w / v% of polysorbate 20, and a pH of approximately 6.3.

[0251] Stability was assessed based on the following key parameters: appearance, high molecular weight protein (HMWP), monomer, and purity. Based on the stability results given in Tables 9 to 22, no trend changes or only minor trend changes were observed in the key parameters related to chemical stability (HMWP, monomer, and purity) and physical stability (appearance) during storage under long-term storage conditions (5°C ± 3°C) and accelerated storage conditions (25°C ± 2°C).

[0252] In summary, the results show that these compositions are chemically and physically stable over time at 5°C and 25°C.

[0253] Analysis program 1. Pharmacopoeia Analysis Procedure The following pharmacopoeia analysis procedures are used to control drug products.

[0254] Appearance According to the European and Japanese pharmacopoeias, the appearance of drug products is determined by visual inspection.

[0255] pH pH was measured using potentiometric assays performed according to the European, American, and Japanese pharmacopoeias.

[0256] 2. Non-Pharmacopoeia Analysis Procedures The following non-pharmacographa analytical procedures are used to control drug products.

[0257] 2.1 Content determined by SE-HPLC The content was determined by SE-HPLC using isocratic elution on a size exclusion column and subsequent UV detection. The content was calculated using the area of ​​the high molecular weight protein (HMWP) peak and monomer peak relative to the area of ​​the HMWP peak and monomer peak of a reference substance with a known concentration, and expressed in mg / ml.

[0258] 2.2 Monomers determined by SE-HPLC The monomer content was determined by SE-HPLC using isocratic elution on a size exclusion column followed by UV detection. The monomer peak area relative to the total area was calculated and expressed as a percentage.

[0259] 2.3 High molecular weight proteins (HMWP) determined by SE-HPLC The content of HMWP was determined by SE-HPLC using isocratic elution on a size exclusion column and subsequent UV detection. The HMWP peak area relative to the total area was calculated and expressed as a percentage.

[0260] 2.4 Purity determined by CE-SDS Purity was defined as the area of ​​the main peak measured by capillary electrophoresis (CE-SDS) in the presence of sodium dodecyl sulfate. Purity was determined by CE-SDS under non-reducing conditions and UV detection. Purity was calculated as the main peak area relative to the total area, expressed as a percentage.

[0261] Stability data for compositions containing mAb1 in the concentration range of 1-100 mg / ml The stability of compositions containing 1, 2, 5, 11.25, 57.5 and 100 mg / mL mAb1 was tracked under long-term storage conditions (5℃ ± 3℃) and accelerated storage conditions (25℃ ± 2℃).

[0262] Compositions containing mAb1 in the range of 1 mg / ml to 100 mg / ml showed considerable stability. The compositions are chemically and physically stable.

[0263] Stability data for 1 mg / ml mAb1 Table 9 reports the stability data of the composition containing 1 mg / ml mAb1 under long-term storage conditions, while Table 10 reports its stability data under accelerated storage conditions.

[0264] Table 9: Stability data of 1 mg / ml mAb1 at 5℃±3℃

[0265] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Table 10: Stability data of 1 mg / ml mAb1 at 25℃±2℃

[0266] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Stability data for 2.0 mg / ml mAb1 Table 11 reports the stability data of 2.0 mg / ml mAb1 under long-term storage conditions, while Table 12 reports its stability data under accelerated storage conditions.

[0267] Table 11: Stability data of 2.0 mg / ml mAb1 at 5℃±3℃

[0268] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Table 12: Stability data of 2.0 mg / ml mAb1 at 25℃±2℃

[0269] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Stability data for 5.0 mg / ml mAb1 Table 13 reports the stability data of 5.0 mg / ml mAb1 under long-term storage conditions, while Table 14 reports its stability data under accelerated storage conditions.

[0270] Table 13: Stability data of 5.0 mg / ml mAb1 at 5℃±3℃

[0271] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Table 14: Stability data of 5.0 mg / ml mAb1 at 25℃±2℃

[0272] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Stability data for 11.3 mg / ml mAb1 Table 15 reports the stability data of 11.3 mg / ml mAb1 under long-term storage conditions, while Table 16 reports its stability data under accelerated storage conditions.

[0273] Table 15: Stability data of 11.3 mg / ml mAb1 at 5℃±3℃

[0274] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Table 16: Stability data of 11.3 mg / ml mAb1 at 25℃±2℃

[0275] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Stability data for 25.0 mg / ml mAb1 Table 17 reports the stability data of 25.0 mg / ml mAb1 under long-term storage conditions, while Table 18 reports its stability data under accelerated storage conditions.

[0276] Table 17: Stability data of 25.0 mg / ml mAb1 at 5℃±3℃

[0277] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Table 18: Stability data of 25.0 mg / ml mAb1 at 25℃±2℃

[0278] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Stability data for 57.5 mg / ml mAb1 Table 19 reports the stability data of 57.5 mg / ml mAb1 under long-term storage conditions, while Table 20 reports its stability data under accelerated storage conditions.

[0279] Table 19: Stability data of 57.5 mg / ml mAb1 at 5℃±3℃

[0280] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Table 20: Stability data of 57.5 mg / ml mAb1 at 25℃±2℃

[0281] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Stability data for 100 mg / ml mAb1 Table 21 reports the stability data of 100 mg / ml mAb1 under long-term storage conditions, while Table 22 reports its stability data under accelerated storage conditions.

[0282] Table 21: Stability data of 100 mg / ml mAb1 at 5℃±3℃

[0283] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. Table 22: Stability data of 100 mg / ml mAb1 at 25℃±2℃

[0284] 1 The criteria for this product are: a clear or nearly clear, colorless or nearly colorless liquid that is substantially free of particles. While certain features of the invention have been set forth and described herein, many modifications, substitutions, alterations, and equivalents will now occur to those skilled in the art. Therefore, it should be understood that all such modifications and alterations falling within the true scope of the invention are intended to be covered by the appended claims.

Claims

1. A bispecific antibody for treating hemophilia A with or without an inhibitor, wherein the bispecific antibody comprises An anti-FIX(a) antibody or its antigen-binding fragment capable of binding to FIX (SEQ ID NO:1) and / or its activated form (FIXa), comprising a heavy chain and a light chain, and Anti-FX(a) antibodies or antigen-binding fragments thereof capable of binding to FX (SEQ ID NO:2) and / or its activated form (FXa), comprising heavy and light chains, in The heavy chain of the anti-FIX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:3, 4 and 5, respectively, and The light chain of the anti-FIX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:8, 9 and 10, respectively, and The heavy chain of the anti-FX(a) antibody or its antigen-binding fragment contains the CDR1-3 sequences represented by SEQ ID NO:13, 14 and 15, respectively, and The light chain of the anti-FX(a) antibody or its antigen-binding fragment comprises the CDR1-3 sequences represented by SEQ ID NO:18, 19 and 20, respectively, and The bispecific antibody will be administered subcutaneously to human patients in a composition containing the bispecific antibody. The loading dose includes - Approximately 9 mg of the bispecific antibody was administered to patients weighing 5 kg to <15 kg, or - Approximately 29 mg of the bispecific antibody was administered to patients weighing 15 kg to <45 kg, or - Approximately 66 mg of the described bispecific antibody, administered to patients weighing 45 kg or more. Maintenance dose includes - Approximately 4 mg of the bispecific antibody, administered every two weeks to patients weighing 5 kg to <15 kg, or - Approximately 9 mg of the bispecific antibody, administered every two weeks to patients weighing 15 kg to <45 kg, or - Approximately 20 mg of the bispecific antibody, administered every two weeks to patients weighing 45 kg or more. The first maintenance dose is administered two weeks after the loading dose.

2. The bispecific antibody for treating hemophilia A with or without an inhibitor, as described in claim 1, wherein... The anti-FIX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:6 and a light chain variable domain represented by SEQ ID NO:11, and wherein The anti-FX(a) antibody or its antigen-binding fragment comprises a heavy chain variable domain represented by SEQ ID NO:16 and a light chain variable domain represented by SEQ ID NO:

21.

3. The bispecific antibody for treating hemophilia A with or without an inhibitor, as described in claim 2, wherein... The heavy chain of the anti-FIX(a) antibody comprises SEQ ID NO:7, and the light chain of the anti-FIX(a) antibody comprises SEQ ID NO:

12. The heavy chain of the anti-FX(a) antibody comprises SEQ ID NO:17, and the light chain of the anti-FX(a) antibody comprises SEQ ID NO:

22.

4. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the preceding claims, wherein the bispecific antibody is mAb1.

5. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the preceding claims, wherein a steady-state plasma concentration of the bispecific antibody is provided in the range of about 2 µg / mL to about 18 µg / mL, such as 3 to 9 µg / mL, such as 6.5 µg / mL.

6. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the preceding claims, wherein the loading dose comprises -9 mg of the bispecific antibody, administered to patients weighing 5 kg to <15 kg, or -29 mg of the bispecific antibody, administered to patients weighing 15 kg to <45 kg, or -66 mg of the bispecific antibody, administered to patients weighing 45 kg or more. Maintenance dose includes -4 mg of the bispecific antibody, administered every two weeks to patients weighing 5 kg to <15 kg, or -9 mg of the bispecific antibody, administered every two weeks to patients weighing 15 kg to <45 kg, or - 20 mg of the bispecific antibody was administered every two weeks to patients weighing 45 kg or more.

7. A bispecific antibody for treating hemophilia A with or without an inhibitor, wherein the bispecific antibody is mAb1, and mAb1 will be administered subcutaneously to human patients in a composition containing mAb1. The loading dose includes -9 mg mAb1, administered to patients weighing 5 kg to <15 kg, or -29 mg mAb1, administered to patients weighing 15 kg to <45 kg, or -66 mg mAb1, administered to patients weighing 45 kg or more. Maintenance dose includes -4 mg mAb1, administered every two weeks to patients weighing 5 kg to <15 kg, or -9 mg mAb1, administered every two weeks to patients weighing 15 kg to <45 kg, or - 20 mg of mAb1, administered every two weeks to patients weighing 45 kg or more. The first maintenance dose is administered two weeks after the loading dose.

8. The bispecific antibody for treating hemophilia A with or without an inhibitor, as claimed in claim 7, wherein a steady-state plasma concentration of mAb1 is provided in the range of 3 to 9 µg / mL, such as 6.5 µg / mL.

9. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the preceding claims, wherein the treatment is a preventative treatment.

10. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the preceding claims, wherein the treatment reduces the frequency of bleeding episodes in patients with hemophilia A with or without an inhibitor.

11. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the preceding claims, wherein the composition comprises about 150 mM of L-arginine hydrochloride or L-arginine, about 20 mM of L-histidine, and a surfactant, such as polysorbate 20 or polysorbate 80, at a pH of about 6.

3.

12. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the preceding claims, wherein the composition comprises about 150 mM of L-arginine hydrochloride or L-arginine, about 20 mM of L-histidine, and about 0.02 w / v% of polysorbate 20 or polysorbate 80, at a pH of about 6.

3.

13. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the preceding claims, wherein the composition comprises about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, and about 0.02 w / v% of polysorbate 20, at a pH of about 6.

3.

14. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the preceding claims, wherein the composition comprises 150 mM of L-arginine hydrochloride, 20 mM of L-histidine, and 0.02 w / v% of polysorbate 20, at a pH of 6.

3.

15. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any one of the preceding claims, wherein the composition comprises the bispecific antibody at a concentration of about 2 mg / ml, about 5 mg / ml, about 11.25 mg / ml, about 25 mg / ml, or about 57.5 mg / ml.

16. A bispecific antibody for treating hemophilia A with or without an inhibitor, according to any of the preceding claims, wherein the treatment provides an annualized bleeding rate of 0, 1, 2, 3 or 4.

17. A pharmaceutical composition comprising a bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, and about 0.02 w / v% of polysorbate 20, at a pH of about 6.

3.

18. The pharmaceutical composition of claim 17, wherein the composition comprises about 2 mg / mL to about 57.5 mg / mL of a bispecific antibody mAb1, about 150 mM of L-arginine hydrochloride, about 20 mM of L-histidine, and about 0.02 w / v% of polysorbate 20, at a pH of about 6.

3.

19. A kit comprising a) a pharmaceutical composition comprising a bispecific antibody according to any one of claims 1-16; and b) instructions for subcutaneous administration of the pharmaceutical composition once every two weeks for the treatment of hemophilia A with or without an inhibitor according to any of the preceding claims.

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