Combination therapy with anti-factor XI / factor XIA antibodies

Abstract

The present disclosure relates to dosage regimens of combination therapies comprising anti-Factor XI and / or Activating Factor XI (Factor XIa) antibodies or antigen-binding fragments thereof, e.g., for the treatment of thromboembolic disorders or related conditions.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 355,319, filed on June 24, 2022, the disclosure of which is hereby incorporated by reference in its entirety for all purposes. Technical Field

[0003] The present disclosure relates to dosage regimens for combination therapies comprising anti-factor XI and / or activated factor XI (factor XIa) antibodies or antigen-binding fragments thereof, e.g., for treating thromboembolic disorders or related conditions. Background Art

[0004] There is a high unmet medical need for safer therapies that reduce thromboembolic complications (e.g., stroke, systemic embolism, cognitive decline, and death), have similar or improved efficacy as demonstrated by existing therapies, and have a lower risk of bleeding.

[0005] Factor XI (FXI) is a serine protease that plays a role in both the intrinsic and extrinsic coagulation pathways. Factor XI exists as a homodimer in the zymogen form; after the peptide bond at R369-I370 is broken, factor XI is activated (factor XIa, FXIa). FXI plays a minor role in normal hemostasis in a high tissue factor environment, but plays a key role in thrombosis. Hereditary factor XI deficiency is associated with a reduced incidence of ischemic stroke and venous thromboembolic events (Salomon et al. (2008); Salomon et al. (2011) Thromb Haemost.; 105:269-73). Bleeding manifestations in subjects with factor XI deficiency are uncommon, usually mild, caused by injury or trauma, and rarely affect vital organs (Salomon et al. (2011)).

[0006] Antibodies that bind to factor XI and / or factor XIa have been studied. For example, WO 2016 / 207858 describes one such anti-factor XI and / or factor XIa antibody, which is disclosed as antibody 1 in Table 1 herein. The present disclosure adds to these developments and provides further clinical methods, including combined dosage regimens, to treat patients with specific thromboembolic diseases with the desired safety and efficacy. Summary of the invention

[0007] The present disclosure provides combination therapies with anti-Factor XI and / or Factor XIa antibodies or antigen-binding fragments thereof, or pharmaceutical formulations comprising the same.

[0008] Thus, in one aspect, provided herein is a method for reversing the anticoagulant effect of an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or antigen-binding fragment thereof in a subject, wherein the isolated antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 150 mg, wherein the method comprises administering to the subject a therapeutically effective amount of recombinant activated factor VII (rFVIIa), thereby reversing the anticoagulant effect.

[0009] In some embodiments, the rFVIIa of the therapeutically effective amount is a low dose. In certain embodiments, the rFVIIa of the therapeutically effective amount is between 0.5 and 1 μg / mL. In some embodiments, the rFVIIa of the therapeutically effective amount is administered once. In some embodiments, the rFVIIa of the therapeutically effective amount is administered more than once. In some embodiments, the rFVIIa of the therapeutically effective amount is administered as needed.

[0010] In some embodiments, relative to the coagulation time (CT) before the administration of a therapeutically effective amount, the administration of a therapeutically effective amount of rFVIIa prolongs CT. In some embodiments, relative to the clot formation time (CFT) before the administration of a therapeutically effective amount, the administration of a therapeutically effective amount of rFVIIa prolongs CFT. In certain embodiments, the CT or CFT is determined by rotational thromboelastometry (ROTEM) in whole blood assays. In certain embodiments, the whole blood assay is ex vivo or in vitro assays.

[0011] In certain embodiments, the method further comprises applying to the subject one of: (i) fluid replacement using colloids, crystalloids, human plasma, or plasma proteins such as albumin; (ii) transfusion of concentrated red blood or whole blood; (iii) administration of fresh frozen plasma (FFP), prothrombin complex concentrate (PCC), activated PCC (APCC), such as a Factor VIII inhibitor, or (iv) an anti-idiotypic antibody to an isolated anti-FXI and / or anti-FXIa antibody or an antigen-binding fragment thereof.

[0012] In another aspect, provided herein is a method of treating a disease or condition in a subject in need thereof, the method comprising administering to the subject a dose of about 150 mg of an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or antigen-binding fragment thereof, and administering (a) about 1 mg / mL aspirin or (b) about 3 µM ticagrelor.

[0013] In some embodiments, the aspirin or ticagrelor is administered prior to administration of the antibody or antigen-binding fragment thereof.

[0014] In some embodiments, the aspirin or ticagrelor is administered concurrently with administration of the antibody or antigen-binding fragment thereof.

[0015] In some embodiments, the aspirin or ticagrelor is administered after administration of the antibody or antigen-binding fragment thereof.

[0016] In some embodiments, compared with subjects treated with aspirin or ticagrelor alone, the administration of the antibody or its antigen-binding fragment does not affect the platelet aggregation of the subject. In certain embodiments, the platelet aggregation is measured by impedance platelet aggregation assay. In certain embodiments, the platelet aggregation is induced by collagen, 5'-adenosine diphosphate (ADP) or thrombin receptor activated peptide-6 (TRAP-6). In certain embodiments, the platelet aggregation is determined in vitro or in vitro.

[0017] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) in SEQ ID NO: 9 or 29, which comprises complementarity determining regions HCDR1, HCDR2 and HCDR3; and a light chain variable region (VL) in SEQ ID NO: 19 or 39, which comprises complementarity determining regions LCDR1, LCDR2, LCDR3.

[0018] In some embodiments of the above aspects, the antibody or antigen-binding fragment thereof comprises:

[0019] i. heavy chain variable region CDR1 of SEQ ID NO: 23; heavy chain variable region CDR2 of SEQ ID NO: 24; heavy chain variable region CDR3 of SEQ ID NO: 25; light chain variable region CDR1 of SEQ ID NO: 33; light chain variable region CDR2 of SEQ ID NO: 34; and light chain variable region CDR3 of SEQ ID NO: 35;

[0020] ii. heavy chain variable region CDR1 of SEQ ID NO: 26; heavy chain variable region CDR2 of SEQ ID NO: 27; heavy chain variable region CDR3 of SEQ ID NO: 28; light chain variable region CDR1 of SEQ ID NO: 36; light chain variable region CDR2 of SEQ ID NO: 37 (or the amino acid sequence of KNY); and light chain variable region CDR3 of SEQ ID NO: 38;

[0021] iii. the heavy chain variable region CDR1 of SEQ ID NO: 43; the heavy chain variable region CDR2 of SEQ ID NO: 44; the heavy chain variable region CDR3 of SEQ ID NO: 45; the light chain variable region CDR1 of SEQ ID NO: 47; the light chain variable region CDR2 of SEQ ID NO: 37 (or the amino acid sequence of KNY); and the light chain variable region CDR3 of SEQ ID NO: 15; or

[0022] iv. the heavy chain variable region CDR1 of SEQ ID NO: 46; the heavy chain variable region CDR2 of SEQ ID NO: 4; the heavy chain variable region CDR3 of SEQ ID NO: 5; the light chain variable region CDR1 of SEQ ID NO: 33; the light chain variable region CDR2 of SEQ ID NO: 14; and the light chain variable region CDR3 of SEQ ID NO: 15.

[0023] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) selected from the group consisting of SEQ ID NOs: 9, 29, and a VH with 90% identity thereof; and a light chain variable region (VL) selected from the group consisting of SEQ ID NOs: 19, 39, and a VL with 90% identity thereof.

[0024] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) selected from the group consisting of SEQ ID NOs: 9 and 29; and a light chain variable region (VL) selected from the group consisting of SEQ ID NOs: 19 and 39.

[0025] In some embodiments, the antibody comprises a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 31, 11, and a heavy chain with 90% identity thereof; and a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 41, 21, and a light chain with 90% identity thereof. In certain embodiments, the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 31, and a light chain comprising an amino acid sequence of SEQ ID NO: 41.

[0026] In some embodiments, the antibody is a human monoclonal antibody.

[0027] In some embodiments, the antibody is of human IgG1 isotype.

[0028] In some embodiments, the antibody comprises D265A and P329A substitutions in the Fc domain.

[0029] In some embodiments, the antibody or antigen-binding fragment thereof is administered intravenously.

[0030] In some embodiments, the antibody or antigen-binding fragment thereof is administered subcutaneously.

[0031] In some embodiments, the antibody or antigen binding fragment thereof is administered in a drug delivery formulation comprising a histidine buffer at a concentration of about 20 mM. In some embodiments, the antibody or antigen binding fragment thereof is administered in a drug delivery formulation comprising sucrose at a concentration of about 220 mM. In some embodiments, the antibody or antigen binding fragment thereof is administered in a drug delivery formulation comprising polysorbate 20 at a concentration of about 0.04%. In some embodiments, the antibody or antigen binding fragment thereof is administered in a drug delivery formulation at a pH of 5.5.

[0032] In some embodiments, when the antibody or antigen-binding fragment thereof is administered in an intravenous drug delivery formulation, the intravenous drug delivery formulation further comprises about 5% glucose.

[0033] In some embodiments, the subject has or is at risk of having a thromboembolic disorder. In some embodiments, the thromboembolic disorder is selected from the group consisting of atrial fibrillation or atrial flutter, transient ischemic attack, ischemic stroke, thromboembolic stroke, hemorrhagic stroke, venous thromboembolism (VTE), pediatric VTE, systemic embolism, non-central nervous system embolism, myocardial infarction, deep vein thrombosis, severe protein S deficiency, cerebrovascular accident and cancer.

[0034] In some embodiments, the first dose of the antibody or antigen-binding fragment thereof is administered intravenously and the second dose of the antibody or antigen-binding fragment is administered subcutaneously. In certain embodiments, the method further comprises administering one or more additional doses of the antibody or antigen-binding fragment thereof subcutaneously after the second dose.

[0035] In some embodiments, the antibody or antigen-binding fragment thereof is administered about once a month.

[0036] Other embodiments and details of the present disclosure are presented below. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1A-1B Depicted in collagen ( Figure 1A ) or TRAP-6 ( Figure 1B ) In vitro platelet aggregation in donor whole blood supplemented with vehicle, Antibody 1, or abciximab after induction.

[0038] Figure 2A-2BFigure 2 shows the effect of combination of antibody 1 and aspirin (ASA) on the expression of 3.2 µg / mL collagen ( Figure 2A ) or 0.5 mM arachidonic acid ( Figure 2B )-induced whole blood platelet aggregation assay in vitro. Error bars represent one standard deviation.

[0039] Figure 3A-3B The combination of Antibody 1 and Ticagrelor is shown in Figure 5. Figure 3A ) or 32 μM TRAP6( Figure 3B )-induced whole blood platelet aggregation assay in vitro. Error bars represent one standard deviation.

[0040] Figure 4A-4B Figure 1 shows the effect of Antibody 1 and rFVIIa on the rotational thromboelastometry (ROTEM) parameters of human blood. Coagulation time (CT, provided in seconds) Figure 4A ) and clot formation time (CFT, given in seconds) ( Figure 4B ). Values ​​are shown as mean and standard deviation percentage values ​​represent differences compared to vehicle alone (baseline). Dashed lines delineate normal reference intervals. * indicates p < 0.05 relative to vehicle alone (baseline). DETAILED DESCRIPTION

[0041] definition

[0042] To facilitate understanding of the present invention, certain terms and phrases are defined below.

[0043] As used herein, the terms "a" and "an" mean "one or more" and include a plural number unless the context is inappropriate.

[0044] As used herein, the terms "FXI protein", "FXI antigen" and "FXI" are used interchangeably and refer to factor XI proteins in different species. Factor XI is mammalian plasma coagulation factor XI, a glycoprotein present in human plasma as a zymogen at a concentration of 25-30 nM, which participates in the intrinsic pathway of blood coagulation when converted to an active serine protease by limited proteolysis.

[0045] The terms "FXIa protein", "FXIa antigen" and "FXIa" are used interchangeably and refer to activated FXI proteins in different species. Enzyme factor XI is converted into its active form, factor Xla (FXIa) via the contact phase of blood coagulation or via thrombin-mediated platelet surface activation. During this activation of factor XI, the internal peptide bond in each of the two chains is cleaved to produce activated factor Xla, a serine protease consisting of two heavy chains and two light chains bound together by disulfide bonds. This serine protease FXIa converts factor IX into IXa, which then activates factor X (Xa). Xa can then mediate factor II / thrombin activation. For example, human FXI has the sequence listed in Table 1 (SEQ ID NO: 1), and has been described in previous reports and literature (Mandle RJ Jr et al. (1979) Blood; 54 (4): 850; NCBI reference sequence: AAA51985).

[0046] In the context of the present disclosure, the terms "FXI" and "FXIa" (and the like) include mutants and variants of native FXI and FXIa proteins, respectively, which have substantially the same amino acid sequence as the native primary structure (amino acid sequence) described in the above reports.

[0047] The terms "catalytic domain", "serine protease catalytic domain" and similar terms used herein refer to amino acids Ile370 to Val607, as counted from Glu1 at the N-terminus of the circulating mature protein. It can also be described as residues 388-625 at the C-terminus of FXI. As used herein, the term "active site" refers to the catalytic triad consisting of amino acids His413, Asp462 and Ser557. (Bane and Gailani (2014) Drug Disc. 19(9)).

[0048] As used herein, the term "antibody" means a complete antibody and any antigen-binding fragment thereof (e.g., "antigen-binding portion") or single chain. A complete antibody is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region consists of three domains CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain CL. The VH and VL regions can be further subdivided into highly variable regions called complementary determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL 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 variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. In some specific aspects, the antibody can be a monoclonal antibody, a human antibody, a humanized antibody, a camel antibody, or a chimeric antibody. The antibody can be of any isotype (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass.

[0049] The CDRs of an antigen binding site can be determined by the methods described in Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991), Chothia et al., J. Mol. Biol. 196:901-917 (1987) and MacCallum et al., J. Mol. Biol. 262:732-745 (1996). When compared to each other, CDRs determined according to these definitions typically include overlapping or subsets of amino acid residues. In certain embodiments, the term "CDR" is a CDR as defined by MacCallum et al., J. Mol. Biol. 262:732-745 (1996) and Martin A., Protein Sequence and Structure Analysis of Antibody Variable Domains, Antibody Engineering, Kontermann and Dubel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001). In certain embodiments, the term "CDR" is a CDR as defined by Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991). In certain embodiments, the heavy chain CDR and light chain CDR of an antibody are defined using different conventions. For example, in certain embodiments, the heavy chain CDR is defined according to MacCallum (supra), while the light chain CDR is defined according to Kabat (supra). CDRH1, CDRH2 and CDRH3 represent heavy chain CDRs, and CDRL1, CDRL2 and CDRL3 represent light chain CDRs.

[0050] As used herein, the term "drug delivery formulation" or "intravenous drug delivery formulation" refers to a pharmaceutical formulation comprising an active agent in combination with an inert or active carrier, making the composition particularly suitable for in vivo or ex vivo diagnostic or therapeutic use.

[0051] As used herein, the terms "subject" and "patient" refer to an organism to be treated by the methods and compositions described herein. Such organisms preferably include, but are not limited to, mammals (e.g., rodents, monkeys, equines, bovines, porcines, primates, canines, felines, etc.), and more preferably include humans. In certain embodiments, the subject is a human. As used herein, "primate subject" includes humans and non-human primates. In certain embodiments, the subject is a baboon model of thrombosis, such as described in Gruber et al., Blood, February 15, 1989; 73(3): 639-42 and Crosby et al., Arterioscler Thromb Vasc Biol, July 2013; 33(7): 1670-8.

[0052] "Thromboembolic disorder" or similar terms as used herein refers to any number of conditions or diseases in which the intrinsic and / or common coagulation pathways are abnormally activated or are not naturally inactivated (e.g., without treatment). These conditions include, but are not limited to, thromboembolic stroke and other types of ischemic stroke, atrial fibrillation, prevention of stroke in atrial fibrillation (SPAF), deep vein thrombosis, venous thromboembolism, and pulmonary embolism. These may also include the prevention and treatment of catheter-related thrombosis (e.g., Hickman catheters in tumor patients), where the catheter thrombi, and extracorporeal membrane oxygenation (ECMO), where the tubing and oxygenation membrane form clots.

[0053] As used herein, "thromboembolic disorder" or similar terms may also refer to any number of the following, which the anti-FXI and / or FXIa antibodies or antigen-binding fragments thereof of the present disclosure may be used to prevent or treat:

[0054] - Thromboembolism in subjects with suspected or confirmed cardiac arrhythmias (e.g., paroxysmal, persistent or permanent atrial fibrillation or atrial flutter);

[0055] - Stroke Prevention in Atrial Fibrillation (SPAF), a subpopulation of which is AF patients undergoing percutaneous coronary intervention (PCI);

[0056] - Treatment of acute venous thromboembolic events (VTE) and extended secondary VTE prophylaxis in patients at high risk of bleeding;

[0057] - Venous thromboembolism, where the subject is a pediatric subject (pediatric VTE);

[0058] - secondary prevention of cerebral and cardiovascular events after transient ischemic attack (TIA) or non-disabling stroke and prevention of thromboembolic events in heart failure with sinus rhythm;

[0059] - Hemorrhagic stroke;

[0060] - Left atrial clot formation and thromboembolism in subjects undergoing cardioversion for arrhythmias;

[0061] - Thrombosis before, during and after arrhythmia ablation procedures;

[0062] - Venous thrombosis, which includes but is not limited to the treatment and secondary prevention of deep or superficial vein thrombosis of the lower or upper extremities, abdominal and thoracic vein thrombosis, sinus thrombosis and jugular vein thrombosis;

[0063] - thrombosis on any artificial surface of a vein or artery, such as a catheter, pacemaker lead, synthetic arterial graft; mechanical or biological heart valve or left ventricular assist device;

[0064] - Pulmonary embolism in patients with or without venous thrombosis;

[0065] - Chronic thromboembolic pulmonary hypertension (CTEPH);

[0066] - Arterial thrombosis on ruptured atherosclerotic plaques, thrombosis on intra-arterial prostheses or catheters, and thrombosis in apparently normal arteries, including but not limited to acute coronary syndrome, ST-segment elevation myocardial infarction, non-ST-segment elevation myocardial infarction, unstable angina, stent thrombosis, thrombosis of any artificial surface of the arterial system, and pulmonary artery thrombosis in subjects with or without pulmonary hypertension;

[0067] - Thrombosis and thromboembolism in patients undergoing percutaneous coronary intervention (PCI);

[0068] - Cardioembolic and cryptogenic strokes;

[0069] - Non-central nervous systemic embolism (non-CNS systemic embolism);

[0070] - Thrombosis in patients with invasive and non-invasive cancer malignancies (e.g. CAT);

[0071] - thrombosis of indwelling catheters;

[0072] - Thrombosis and thromboembolism in critically ill patients;

[0073] - Cardiac thrombosis and thromboembolism, including but not limited to cardiac thrombosis following myocardial infarction, cardiac thrombosis associated with conditions such as cardiac aneurysms, myocardial fibrosis, cardiac enlargement and insufficiency, myocarditis and cardiac artificial surfaces;

[0074] - Thromboembolism in patients with valvular heart disease with or without atrial fibrillation;

[0075] - Thromboembolism of mechanical or biological valve prostheses;

[0076] - Thromboembolism in patients with natural or artificial cardiac patches, arterial or venous conduits, after cardiac repair for simple or complex cardiac malformations;

[0077] - venous thrombosis and thromboembolism following knee replacement surgery, hip replacement surgery, and orthopedic, chest or abdominal surgery;

[0078] - Arterial or venous thrombosis following neurosurgery, including intracranial and spinal interventions;

[0079] - Congenital or acquired thrombotic tendencies, including but not limited to factor V Leiden, prothrombin mutation, antithrombin III, protein C and protein S deficiency, factor XIII mutation, familial dysfibrinogenemia, congenital plasminogen deficiency, elevated factor XI levels, sickle cell disease, antiphospholipid syndrome, autoimmune diseases, chronic intestinal diseases, nephrotic syndrome, hemolytic uremic disease, myeloproliferative diseases, disseminated intravascular coagulation, paroxysmal nocturnal hemoglobinuria, and heparin-induced thrombocytopenia;

[0080] - Thrombosis and thromboembolism in chronic kidney disease; and

[0081] -Thrombosis and thromboembolism in patients undergoing hemodialysis and patients receiving extracorporeal membrane oxygenation.

[0082] As used herein, the term "trough" or "trough level" refers to the lowest concentration of a drug reached before the next dose of the drug is administered. In certain embodiments, the inhibition of factor XI / factor XIa at the trough is greater than about 50% (e.g., greater than about 60%, greater than about 70%, greater than about 80%, or greater than about 90%). In certain embodiments, the inhibition of factor XI / factor XIa at the trough is greater than about 80%. In certain embodiments, the inhibition of factor XI / factor XIa at the trough is greater than about 90%.

[0083] The terms "treat / treating / treatment" and other grammatical equivalents as used in this disclosure include alleviating, alleviating, ameliorating, or preventing a disease, disorder, or symptom; preventing other symptoms; ameliorating or preventing the underlying metabolic causes of a symptom; inhibiting a disease or disorder, such as arresting the development of a disease or disorder, alleviating a disease or disorder, causing regression of a disease or disorder, alleviating a disorder caused by a disease or disorder, or stopping the symptoms of a disease or disorder; and are intended to include prevention. The terms also include achieving a therapeutic benefit and / or a preventive benefit. "Therapeutic benefit" means eradicating or ameliorating the underlying condition being treated. In addition, a therapeutic benefit is achieved by eradicating or ameliorating one or more physiological symptoms associated with the underlying condition, thereby observing an improvement in the patient, although the patient may still suffer from the underlying condition.

[0084] In certain embodiments of the methods described herein, the subject has not been treated, i.e., has never received any form of anticoagulant therapy prior to treatment with an anti-factor XI / XIa antibody (e.g., antibody 1) described herein. In certain embodiments of the methods described herein, the subject has received stable treatment with a recommended dose of a novel oral anticoagulant (NOAC), e.g., prior to treatment with an anti-factor XI / XIa antibody (e.g., antibody 1) described herein. In certain embodiments, the subject has received a direct oral anticoagulant (DOAC), e.g., prior to treatment with an anti-factor XI / XIa antibody (e.g., antibody 1) described herein. In certain embodiments, the subject has received a vitamin K antagonist (VKA), e.g., prior to treatment with an anti-factor XI / XIa antibody (e.g., antibody 1) described herein.

[0085] As used herein, the term "vial" refers to a container that holds a pharmaceutical product. In some embodiments, the vial can be a vial, a bag, a pen, or a syringe. In some embodiments, the vial can be a vial, such as a glass vial.

[0086] As used herein, the term "drug product" refers to an anti-Factor XI / XIa antibody described herein, such as Antibody 1 disclosed in Table 1, and excipients, such as histidine buffer, sugars, and polysorbates.

[0087] The term "about" refers to any minimal change in the concentration or amount of a pharmaceutical agent that does not change the efficacy of the pharmaceutical agent in the preparation of the formulation and the treatment of a disease or condition. In certain embodiments, the term "about" may include ±5%, ±10%, or ±15% of a specified numerical value or data point.

[0088] Ranges can be expressed in the present disclosure as from "about" a specific value, and / or to "about" another specific value. When such ranges are expressed, another aspect includes from a specific value and / or to another specific value. Similarly, when a value is expressed as an approximation by using the antecedent "about", it should be understood that the specific value forms another aspect. It should also be understood that the endpoints of each range are significant relative to the other endpoint and are independent of the other endpoint. It should also be understood that multiple values ​​are disclosed in the present disclosure, and each value is also disclosed as "about" the specific value in addition to the value itself. It should also be understood that throughout this application, data is provided in a variety of different formats, and this data represents the scope of any combination of endpoints and starting points and data points. For example, if a specific data point "10" and a specific data point "15" are disclosed, it should be understood that it is considered to disclose greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 and between 10 and 15. It should also be understood that each unit between two specific units is also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0089] Throughout this specification, when compositions are described as having, including or comprising specific components, or when processes and methods are described as having, including or comprising specific steps, it is contemplated that there are additional compositions of the invention that consist essentially of or consist of those components, and that there are processes and methods according to the invention that consist essentially of or consist of those process steps.

[0090] In general, compositions specifying percentages are by weight unless otherwise indicated. In addition, if a variable is not accompanied by a definition, the preceding definition of the variable will prevail.

[0091] Anti-factor XI and / or activated factor XI (factor XIa) antibodies

[0092] In some embodiments, the present disclosure provides a pharmaceutical formulation comprising an antibody that binds to FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), wherein the antibody comprises a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29, the formulation comprises a histidine buffer; a sugar or sugar alcohol; and a polysorbate, and the pH of the formulation is between pH 5.0 and 6.0. In certain embodiments, the antibody comprises a VH having an amino acid sequence of SEQ ID NO: 29.

[0093] In an embodiment, the present disclosure provides a pharmaceutical formulation comprising an antibody that binds to a FXI and / or FXIa protein or an antigen-binding fragment thereof, the pharmaceutical formulation being contained in a vial, wherein the formulation comprises an excess fill volume for completely withdrawing a therapeutically effective amount of an anti-FXI and / or anti-FXIa antibody or an antigen-binding fragment thereof. In certain embodiments, the vial contains a pharmaceutical formulation comprising about 150 mg of an antibody that binds to a FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 9 or 29; a histidine buffer at a concentration of about 20 mM; a sucrose at a concentration of about 220 mM; and a polysorbate-20 at a concentration of about 0.04% (v / v); and the pH of the formulation is about pH 5.5.

[0094] In an embodiment, the present disclosure provides an intravenously delivered pharmaceutical formulation comprising about 1.5 mg of an antibody or antigen-binding fragment thereof that binds to FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 9 or 29; a histidine buffer at a concentration of about 0.20 mM; sucrose at a concentration of about 2.20 mM; polysorbate-20 at a concentration of about 0.0004% (v / v) and a diluent (e.g., 5% dextrose in water (D5W)); and the pH of the formulation is about pH 5.5.

[0095] The present disclosure also provides pharmaceutical formulations of antibodies that specifically bind to FXI and / or FXIa proteins, wherein the antibody comprises a VH CDR having an amino acid sequence of any one of the VH CDRs listed in Table 1 below, the formulation comprises a histidine buffer; a sugar or sugar alcohol; and a polysorbate; and the pH of the formulation is between pH 5.0 and 6.0. In particular, the present disclosure provides pharmaceutical formulations of antibodies that specifically bind to FXI and / or FXIa proteins (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), wherein the antibody comprises one, two, three or more VH CDRs having an amino acid sequence of any one of the VH CDRs listed in Table 1 below (or alternatively, consists thereof), the formulation comprises a histidine buffer; a sugar or sugar alcohol; and a polysorbate; and the pH of the formulation is between pH 5.0 and 6.0. (See PCT International Patent Application No. PCT / IB2016 / 053790 filed on June 24, 2016 and published as WO2016 / 207858, which is hereby incorporated by reference in its entirety).

[0096] In some embodiments, the present disclosure provides a pharmaceutical formulation of an antibody that specifically binds to a FXI / FXIa protein, the antibody comprising a light chain variable domain (VL) having an amino acid sequence of SEQ ID NO: 19 or 39, for use in the methods described herein (e.g., a method for treating a subject having or at risk of having a thromboembolic disorder), the formulation comprising a histidine buffer; a sugar or sugar alcohol; and a polysorbate; and the pH of the formulation is between pH 5.0 and 6.0. In certain embodiments, the antibody comprises a VL having an amino acid sequence of SEQ ID NO: 39.

[0097] In an embodiment, the present disclosure provides a pharmaceutical formulation comprising an antibody that binds to a FXI and / or FXIa protein or an antigen-binding fragment thereof, the pharmaceutical formulation being contained in a vial, wherein the formulation comprises an excess fill volume for completely withdrawing a therapeutically effective amount of an anti-FXI and / or anti-FXIa antibody or an antigen-binding fragment thereof. In certain embodiments, the vial contains a pharmaceutical formulation comprising about 150 mg of an antibody that binds to a FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 19 or 39; a histidine buffer at a concentration of about 20 mM; a sucrose at a concentration of about 220 mM; and a polysorbate-20 at a concentration of about 0.04% (v / v); and the pH of the formulation is about pH 5.5.

[0098] In an embodiment, the present disclosure provides an intravenously delivered pharmaceutical formulation comprising about 1.5 mg of an antibody or antigen-binding fragment thereof that binds to FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 19 or 39; a histidine buffer at a concentration of about 0.20 mM; sucrose at a concentration of about 2.20 mM; polysorbate-20 at a concentration of about 0.0004% (v / v) and a diluent (e.g., 5% dextrose in water (D5W)); and the pH of the formulation is about pH 5.5.

[0099] The present disclosure also provides pharmaceutical formulations of antibodies that specifically bind to FXI and / or FXIa proteins (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), for use in the methods described herein (e.g., methods for treating a subject having or at risk of having a thromboembolic disorder), the antibodies comprising a VL CDR having an amino acid sequence of any one of the VL CDRs listed in Table 1 below; the formulation comprising a histidine buffer; a sugar or sugar alcohol; and a polysorbate; and the pH of the formulation is between pH 5.0 and 6.0. The antibodies that specifically bind to FXIa proteins (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa) may comprise one, two, three or more VL CDRs having an amino acid sequence of any one of the VL CDRs listed in Table 1 below (or alternatively, consist of).

[0100] In an embodiment, the present disclosure provides a pharmaceutical formulation comprising an antibody that binds to a FXI and / or FXIa protein or an antigen-binding fragment thereof, the pharmaceutical formulation being contained in a vial, wherein the formulation comprises an excess fill volume for completely withdrawing a therapeutically effective amount of an anti-FXI and / or anti-FXIa antibody or an antigen-binding fragment thereof. In certain embodiments, the vial contains a pharmaceutical formulation comprising about 150 mg of an antibody that binds to a FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 9 or 29, and a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 19 or 39; a histidine buffer at a concentration of about 20 mM; a sucrose at a concentration of about 220 mM; and a polysorbate-20 at a concentration of about 0.04% (v / v); and the pH of the formulation is about pH 5.5.

[0101] In an embodiment, the present disclosure provides an intravenously delivered pharmaceutical formulation comprising about 1.5 mg of an antibody or antigen-binding fragment thereof that binds to FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 9 or 29, and a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 19 or 39; a histidine buffer at a concentration of about 0.20 mM; sucrose at a concentration of about 2.20 mM; polysorbate-20 at a concentration of about 0.0004% (v / v) and a diluent (e.g., 5% dextrose in water (D5W)); and the pH of the formulation is about pH 5.5.

[0102] In an embodiment, the present disclosure provides a pharmaceutical formulation comprising an antibody that binds to FXI and / or FXIa proteins or antigen-binding fragments thereof, the pharmaceutical formulation being contained in a vial, wherein the formulation comprises an excess fill volume for completely withdrawing a therapeutically effective amount of the anti-FXI and / or anti-FXIa antibody or antigen-binding fragment thereof. In certain embodiments, the vial contains a pharmaceutical formulation comprising about 150 mg of an antibody that binds to FXI and / or FXIa proteins (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 29, and a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 39; a histidine buffer at a concentration of about 20 mM; a sucrose at a concentration of about 220 mM; and a polysorbate-20 at a concentration of about 0.04% (v / v); and the pH of the formulation is about pH 5.5.

[0103] In an embodiment, the present disclosure provides an intravenously delivered pharmaceutical formulation comprising about 1.5 mg of an antibody or antigen-binding fragment thereof that binds to FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 29, and a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 39; a histidine buffer at a concentration of about 0.20 mM; a sucrose at a concentration of about 2.20 mM; a polysorbate-20 at a concentration of about 0.0004% (v / v) and a diluent (e.g., 5% dextrose in water (D5W)); and the pH of the formulation is about pH 5.5.

[0104] In some embodiments, other antibodies for use in the methods described herein (e.g., methods for treating a subject having or at risk for a thromboembolic disorder) include amino acids that have been mutated but have at least 60%, 70%, 80%, 85%, 90%, or 95% CDR region identity with the CDR regions depicted in the sequences set forth in Table 1. In some embodiments, the antibodies include mutant amino acid sequences, wherein no more than 1, 2, 3, 4, or 5 amino acids in the CDR regions have been mutated compared to the CDR regions depicted in the sequences set forth in Table 1.

[0105] Table 1. Examples of FXI / FXIa antibodies, Fabs, and FXI / FXIa proteins

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116] In some embodiments, other antibodies for use in the methods or formulations described herein (e.g., methods for treating a subject having or at risk for a thromboembolic disorder) include antibodies in which the amino acids or nucleic acids encoding the amino acids have been mutated, but have at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% identity to the sequences set forth in Table 1. Some embodiments include mutant amino acid sequences in which no more than 1, 2, 3, 4, or 5 amino acids in the variable region have been mutated compared to the variable region depicted in the sequences set forth in Table 1, while retaining substantially the same antigen binding activity.

[0117] Since each of these antibodies can bind to FXI and / or FXIa, the VH, VL, full-length light chain and full-length heavy chain sequences (amino acid sequences and nucleotide sequences encoding the amino acid sequences) can be "mixed and matched" to generate other FXI and / or FXIa binding antibodies of the present disclosure. Such "mixed and matched" FXI and / or FXIa binding antibodies can be tested using binding assays known in the art (e.g., ELISA and other assays described in the Examples section). When these chains are mixed and matched, the VH sequence from a specific VH / VL pairing should be replaced with a structurally similar VH sequence. Similarly, the full-length heavy chain sequence from a specific full-length heavy chain / full-length light chain pairing should be replaced with a structurally similar full-length heavy chain sequence. Similarly, the VL sequence from a specific VH / VL pairing should be replaced with a structurally similar VL sequence. Similarly, the full-length light chain sequence from a specific full-length heavy chain / full-length light chain pairing should be replaced with a structurally similar full-length light chain sequence.

[0118] Thus, in one aspect, for use in the methods described herein (e.g., methods for treating a subject having or at risk of having a thromboembolic disorder), the present disclosure provides an isolated antibody or antigen-binding fragment thereof having: a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 29, and a light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 19 and 39, wherein the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey, and baboon FXIa). In another aspect, for use in the formulations described herein (e.g., formulations in vials, intravenous drug delivery formulations), the present disclosure provides an isolated antibody or antigen-binding fragment thereof having: a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 29, and a light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 19 and 39, wherein the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey, and baboon FXIa).

[0119] More specifically, in certain aspects, the present disclosure provides an isolated antibody or antigen-binding fragment thereof having a heavy chain variable domain and a light chain variable domain comprising an amino acid sequence selected from SEQ ID NOs: 9 and 29 or 19 and 39, respectively.

[0120] In a specific embodiment for use in the methods described herein (e.g., methods for treating a subject having or at risk for a thromboembolic disorder), the antibody or antigen-binding fragment thereof provided herein that specifically binds to human FXI and / or FXIa contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 19. In a specific embodiment for use in a formulation described herein (e.g., a formulation in a vial, an intravenous drug delivery formulation), the antibody or antigen-binding fragment thereof provided herein that specifically binds to human FXI and / or FXIa contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 19.

[0121] In a specific embodiment for use in the methods described herein (e.g., methods for treating a subject having or at risk for a thromboembolic disorder), the antibody or antigen-binding fragment thereof provided herein that specifically binds to human FXI and / or FXIa contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 29, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 39. In a specific embodiment for use in a formulation described herein (e.g., a formulation in a vial, an intravenous drug delivery formulation), the antibody or antigen-binding fragment thereof provided herein that specifically binds to human FXI and / or FXIa contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 29, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 39.

[0122] In another aspect for use in the methods described herein, the present disclosure provides (i) an isolated antibody having: a full-length heavy chain comprising an amino acid sequence optimized for expression in a mammalian cell selected from the group consisting of SEQ ID NO: 11 or 31, and a full-length light chain comprising an amino acid sequence optimized for expression in a mammalian cell selected from the group consisting of SEQ ID NO: 21 or 41; or (ii) a functional protein comprising an antigen binding portion thereof. More specifically, in certain aspects, the present disclosure provides an isolated antibody or antigen binding fragment thereof having a heavy chain and a light chain comprising an amino acid sequence selected from SEQ ID NO: 11 and 31 or 21 and 41, respectively. In another aspect for use in the formulations described herein, the present disclosure provides (i) an isolated antibody having: a full-length heavy chain comprising an amino acid sequence optimized for expression in a mammalian cell selected from the group consisting of SEQ ID NO: 11 or 31, and a full-length light chain comprising an amino acid sequence optimized for expression in a mammalian cell selected from the group consisting of SEQ ID NO: 21 or 41; or (ii) a functional protein comprising an antigen binding portion thereof. More specifically, in certain aspects, the present disclosure provides an isolated antibody or antigen-binding fragment thereof having a heavy chain and a light chain comprising an amino acid sequence selected from SEQ ID NOs: 11 and 31 or 21 and 41, respectively.

[0123] In a specific embodiment for use in the methods described herein, the antibodies or antigen-binding fragments thereof provided herein that specifically bind to human FXI and / or FXIa contain a heavy chain comprising the amino acid sequence of SEQ ID NO: 11, and a light chain comprising the amino acid sequence of SEQ ID NO: 21. In a specific embodiment for use in the formulations described herein, the antibodies or antigen-binding fragments thereof provided herein that specifically bind to human FXI and / or FXIa contain a heavy chain comprising the amino acid sequence of SEQ ID NO: 11, and a light chain comprising the amino acid sequence of SEQ ID NO: 21.

[0124] In a specific embodiment for use in the methods described herein, an antibody or antigen-binding fragment thereof that specifically binds to human FXI and / or FXIa provided herein contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 41. In a specific embodiment for use in the formulations described herein, an antibody or antigen-binding fragment thereof that specifically binds to human FXI and / or FXIa provided herein contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 41.

[0125] As used herein, the terms "complementarity determining region" and "CDR" refer to the amino acid sequences within the variable region of an antibody that confer antigen specificity and binding affinity. Typically, each heavy chain variable region has three CDRs (HCDR1, HCDR2, HCDR3), and each light chain variable region has three CDRs (LCDR1, LCDR2, LCDR3).

[0126] The precise amino acid sequence boundaries of a given CDR can be readily determined using any of a number of well-known schemes, including those described by Kabat et al., (1991), "Sequences of Proteins of Immunological Interest", 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); Lefranc et al., (2003) Dev. Comp. Immunol., 27, 55-77 ("IMGT" numbering scheme); or a "combined" system.

[0127] For example, according to Kabat, the CDR amino acid residues of Antibody 2 in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-66 (HCDR2), and 99-111 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 22-35 (LCDR1), 51-57 (LCDR2), and 90-100 (LCDR3). According to Chothia, the CDR amino acid residues in VH are numbered 26-32 (HCDR1), 52-57 (HCDR2), and 99-111 (HCDR3); and the amino acid residues in VL are numbered 25-33 (LCDR1), 51-53 (LCDR2), and 92-99 (LCDR3). By combining the CDR definitions of Kabat and Chothia, the CDRs consist of amino acid residues 26-35 (HCDR1), 50-66 (HCDR2), and 99-111 (HCDR3) in human VH and amino acid residues 22-35 (LCDR1), 51-57 (LCDR2), and 90-100 (LCDR3) in human VL. By combining the CDR definitions of Kabat and Chothia, the "combined" CDRs consist of amino acid residues 26-35 (HCDR1), 50-66 (HCDR2), and 99-108 (HCDR3) in human VH and amino acid residues 24-38 (LCDR1), 54-60 (LCDR2), and 93-101 (LCDR3) in human VL. As another example, according to IMGT, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 26-33 (HCDR1), 51-58 (HCDR2), and 97-108 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 27-36 (LCDR1), 54-56 (LCDR2), and 93-101 (LCDR3). Table 1 provides exemplary Kabat, Chothia, combination, and IMGT HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 for anti-FXI / FXIa antibodies, e.g., Antibody 2 and Antibody 1. In another aspect, the present disclosure provides FXIa-binding antibodies comprising heavy and light chain CDR1, CDR2, and CDR3 as described in Table 1, or a combination thereof. The amino acid sequence of the VH CDR1 of the antibody is shown in SEQ ID NOs: 3 and 23. The amino acid sequence of the VH CDR2 of the antibody is shown in SEQ ID NOs: 4 and 24. The amino acid sequence of the VH CDR3 of the antibody is shown in SEQ ID NOs: 5 and 25. The amino acid sequence of the VL CDR1 of the antibody is shown in SEQ ID NOs: 13 and 33.The amino acid sequence of the VL CDR2 of the antibody is shown in SEQ ID NOs: 14 and 34. The amino acid sequence of the VL CDR3 of the antibody is shown in SEQ ID NOs: 15 and 35. These CDR regions are delineated using the Kabat system.

[0128] Alternatively, as defined using the Chothia system (Al-Lazikani et al., (1997) JMB 273, 927-948), the amino acid sequence of the VH CDR1 of the antibody is shown in SEQ ID NOs: 6 and 26. The amino acid sequence of the VH CDR2 of the antibody is shown in SEQ ID NOs: 7 and 27. The amino acid sequence of the VH CDR3 of the antibody is shown in SEQ ID NOs: 8 and 28. The amino acid sequence of the VL CDR1 of the antibody is shown in SEQ ID NOs: 16 and 36. The amino acid sequence of the VL CDR2 of the antibody is shown in SEQ ID NOs: 17 and 37 or has the amino acid sequence of KNY. The amino acid sequence of the VL CDR3 of the antibody is shown in SEQ ID NOs: 18 and 38.

[0129] Alternatively, as defined using a combinatorial system, the amino acid sequence of the VH CDR1 of the antibody is shown in SEQ ID NO: 46. The amino acid sequence of the VH CDR2 of the antibody is shown in SEQ ID NO: 4. The amino acid sequence of the VH CDR3 of the antibody is shown in SEQ ID NO: 5. The amino acid sequence of the VL CDR1 of the antibody is shown in SEQ ID NO: 33. The amino acid sequence of the VL CDR2 of the antibody is shown in SEQ ID NO: 14. The amino acid sequence of the VL CDR3 of the antibody is shown in SEQ ID NO: 15.

[0130] Alternatively, as defined using the IMGT numbering scheme, the amino acid sequence of the VH CDR1 of the antibody is shown in SEQ ID NO: 43. The amino acid sequence of the VH CDR2 of the antibody is shown in SEQ ID NO: 44. The amino acid sequence of the VH CDR3 of the antibody is shown in SEQ ID NO: 45. The amino acid sequence of the VL CDR1 of the antibody is shown in SEQ ID NO: 47. The amino acid sequence of the VL CDR2 of the antibody is shown in SEQ ID NO: 37 (or the amino acid sequence of KNY). The amino acid sequence of the VL CDR3 of the antibody is shown in SEQ ID NO: 15.

[0131] Given that each of these antibodies can bind to FXI and / or FXIa, and that antigen binding specificity is provided primarily by the CDR1, 2, and 3 regions, the VH CDR1, 2, and 3 sequences and the VL CDR1, 2, and 3 sequences can be "mixed and matched" (e.g., CDRs from different antibodies can be mixed and matched, although each antibody preferably contains VH CDR1, 2, and 3 and VL CDR1, 2, and 3 to generate other FXI and / or FXIa binding molecules of the present disclosure). Such "mixed and matched" FXI and / or FXIa binding antibodies can be tested using binding assays known in the art and those described in the Examples (e.g., ELISA, SET, BIACORE™ assays). When mixing and matching VH CDR sequences, the CDR1, CDR2, and / or CDR3 sequences from a particular VH sequence should be replaced with CDR sequences of similar structure. Similarly, when mixing and matching VL CDR sequences, the CDR1, CDR2, and / or CDR3 sequences from a particular VL sequence should be replaced with CDR sequences of similar structure. It will be apparent to one of ordinary skill that new VH and VL sequences can be generated by replacing one or more VH and / or VL CDR region sequences with structurally similar sequences from the CDR sequences of the monoclonal antibodies used in the present disclosure set forth herein. In addition to the foregoing, in one embodiment, an antigen-binding fragment of an antibody described herein may comprise VH CDR1, 2, and 3, or VL CDR 1, 2, and 3, wherein the fragment binds to FXI and / or FXIa as a single variable domain. Note that the CDR sequences of Antibody 1 and Antibody 2 are identical.

[0132] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof may have the heavy chain and light chain sequences of the Fab described in Table 1. More specifically, the antibody or antigen-binding fragment thereof may have the heavy chain and light chain sequences of Antibody 2 and Antibody 1.

[0133] In other embodiments of the present disclosure, the antibody or antigen-binding fragment that specifically binds to FXI and / or FXIa comprises a heavy chain variable region CDR1, a heavy chain variable region CDR2, a heavy chain variable region CDR3, a light chain variable region CDR1, a light chain variable region CDR2, and a light chain variable region CDR3, as defined by Kabat and described in Table 1. In other embodiments of the present disclosure, the antibody or antigen-binding fragment that specifically binds to FXI and / or FXIa comprises a heavy chain variable region CDR1, a heavy chain variable region CDR2, a heavy chain variable region CDR3, a light chain variable region CDR1, a light chain variable region CDR2, and a light chain variable region CDR3, as defined by Chothia and described in Table 1. In other embodiments, the antibody or antigen-binding fragment that specifically binds to FXI and / or FXIa comprises a heavy chain variable region CDR1, a heavy chain variable region CDR2, a heavy chain variable region CDR3, a light chain variable region CDR1, a light chain variable region CDR2, and a light chain variable region CDR3, as defined by a combined system and described in Table 1. In other embodiments of the present disclosure, the antibody or antigen-binding fragment that specifically binds to FXI and / or FXIa comprises a heavy chain variable region CDR1, a heavy chain variable region CDR2, a heavy chain variable region CDR3, a light chain variable region CDR1, a light chain variable region CDR2, and a light chain variable region CDR3, as defined by IMGT and described in Table 1.

[0134] In a specific embodiment for use in the methods described herein, the disclosure includes an antibody that specifically binds to FXI and / or FXIa, comprising a heavy chain variable region CDR1 of SEQ ID NO: 3; a heavy chain variable region CDR2 of SEQ ID NO: 4; a heavy chain variable region CDR3 of SEQ ID NO: 5; a light chain variable region CDR1 of SEQ ID NO: 13; a light chain variable region CDR2 of SEQ ID NO: 14; and a light chain variable region CDR3 of SEQ ID NO: 15.

[0135] In a specific embodiment, the present disclosure includes an antibody that specifically binds to FXI and / or FXIa, comprising a heavy chain variable region CDR1 of SEQ ID NO: 23; a heavy chain variable region CDR2 of SEQ ID NO: 24; a heavy chain variable region CDR3 of SEQ ID NO: 25; a light chain variable region CDR1 of SEQ ID NO: 33; a light chain variable region CDR2 of SEQ ID NO: 34; and a light chain variable region CDR3 of SEQ ID NO: 35.

[0136] In a specific embodiment, the present disclosure includes an antibody that specifically binds to FXI and / or FXIa, which comprises a heavy chain variable region CDR1 of SEQ ID NO: 6; a heavy chain variable region CDR2 of SEQ ID NO: 7; a heavy chain variable region CDR3 of SEQ ID NO: 8; a light chain variable region CDR1 of SEQ ID NO: 16; a light chain variable region CDR2 of SEQ ID NO: 17 (or the amino acid sequence of KNY); and a light chain variable region CDR3 of SEQ ID NO: 18.

[0137] In a specific embodiment, the present disclosure includes an antibody that specifically binds to FXI and / or FXIa, which comprises a heavy chain variable region CDR1 of SEQ ID NO: 26; a heavy chain variable region CDR2 of SEQ ID NO: 27; a heavy chain variable region CDR3 of SEQ ID NO: 28; a light chain variable region CDR1 of SEQ ID NO: 36; a light chain variable region CDR2 of SEQ ID NO: 37 (or the amino acid sequence of KNY); and a light chain variable region CDR3 of SEQ ID NO: 38.

[0138] In a specific embodiment, provided herein is an antibody that specifically binds to FXI and / or FXIa, which comprises a heavy chain variable region CDR1 of SEQ ID NO: 43; a heavy chain variable region CDR2 of SEQ ID NO: 44; a heavy chain variable region CDR3 of SEQ ID NO: 45; a light chain variable region CDR1 of SEQ ID NO: 47; a light chain variable region CDR2 of SEQ ID NO: 37 (or the amino acid sequence of KNY); and a light chain variable region CDR3 of SEQ ID NO: 15.

[0139] In a specific embodiment, provided herein is an antibody that specifically binds to FXI and / or FXIa, comprising a heavy chain variable region CDR1 of SEQ ID NO: 46; a heavy chain variable region CDR2 of SEQ ID NO: 4; a heavy chain variable region CDR3 of SEQ ID NO: 5; a light chain variable region CDR1 of SEQ ID NO: 33; a light chain variable region CDR2 of SEQ ID NO: 14; and a light chain variable region CDR3 of SEQ ID NO: 15.

[0140] In certain embodiments, the present disclosure includes antibodies or antigen-binding fragments that specifically bind to FXI and / or FXIa as described in Table 1. In a specific embodiment for the methods described herein, the antibodies or antigen-binding fragments that bind to FXI and / or FXIa are Antibody 2 and Antibody 1.

[0141] As used herein, a human antibody comprises a heavy or light chain variable region or a full-length heavy or light chain, and if the variable region or full-length chain of the antibody is obtained from a system using human germline immunoglobulin genes, the variable region or full-length chain is the "product" or "derived from" a specific germline sequence. Such systems include immunizing transgenic mice carrying human immunoglobulin genes with an antigen of interest or screening a human immunoglobulin gene library displayed on phage with an antigen of interest. Human antibodies that are "products" or "derived from" human germline immunoglobulin sequences can be identified by comparing the amino acid sequence of a human antibody with the amino acid sequence of a human germline immunoglobulin and selecting the human germline immunoglobulin sequence that is closest in sequence to the human antibody sequence (i.e., the greatest % identity).

[0142] A human antibody that is the "product" of or "derived from" a particular human germline immunoglobulin sequence may contain amino acid differences compared to the germline sequence, for example, due to naturally occurring somatic mutations or intentionally introduced site-directed mutations. However, in the VH or VL framework regions, the selected human antibody is typically at least 90% identical in amino acid sequence to the amino acid sequence encoded by the human germline immunoglobulin gene, and contains amino acid residues that identify the human antibody as human when compared to germline immunoglobulin amino acid sequences of other species (e.g., murine germline sequences). In some cases, the amino acid sequence of the human antibody may be at least 60%, 70%, 80%, 90%, or at least 95%, or even at least 96%, 97%, 98%, or 99% identical to the amino acid sequence encoded by the germline immunoglobulin gene.

[0143] Typically, a recombinant human antibody will exhibit no more than 10 amino acid differences in the VH or VL framework regions from the amino acid sequence encoded by a human germline immunoglobulin gene. In certain cases, a human antibody may exhibit no more than 5, or even no more than 4, 3, 2, or 1 amino acid differences from the amino acid sequence encoded by a germline immunoglobulin gene. Examples of human germline immunoglobulin genes include, but are not limited to, the variable domain germline segments described below, as well as DP47 and DPK9.

[0144] Homologous antibody

[0145] In some other embodiments for use in the methods described herein (e.g., methods for treating a subject having or at risk of a thromboembolic disorder), the disclosure provides an antibody or antigen-binding fragment thereof comprising an amino acid sequence homologous to a sequence described in Table 1 (e.g., SEQ ID NO: 29, 31, 39, or 41), and the antibody binds to FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXIa), and retains the desired functional properties of those antibodies described in Table 1, e.g., Antibody 2 and Antibody 1. In specific aspects, such homologous antibodies retain the CDR amino acid sequences described in Table 1 (e.g., Kabat CDRs, Chothia CDRs, IMGT CDRs, or combined CDRs).

[0146] For example, in some embodiments, the present disclosure provides an isolated antibody or a functional antigen-binding fragment thereof, comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 29; the light chain variable domain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 19 and 39; and the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey and baboon FXIa). In one embodiment, the isolated antibody or functional antigen-binding fragment thereof comprises a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 9; the light chain variable domain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 19; and the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey and baboon FXIa). In one embodiment, the isolated antibody or its functional antigen-binding fragment comprises a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 29; the light chain variable domain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 39; and the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey and baboon FXIa). In certain aspects of the present disclosure, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences defined by Kabat, e.g., SEQ ID NOs: 3, 4, 5, 13, 14 and 15, respectively. In certain other aspects of the present disclosure, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences defined by Chothia, such as SEQ ID NOs: 6, 7, 8, 16, 17 (or the amino acid sequence of KNY), and 18, respectively. In certain other aspects, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences defined by the Combinatorial System, such as SEQ ID NOs: 46, 4, 5, 33, 14, and 15, respectively.In certain other aspects, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences defined by IMGT, such as SEQ ID NOs: 43, 44, 45, 47, 37 (or the amino acid sequence of KNY) and 15, respectively.

[0147] In other embodiments for use in the methods described herein, the VH and / or VL amino acid sequences may be 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequences listed in Table 1. In other embodiments for use in the formulations described herein, the VH and / or VL amino acid sequences may be 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequences listed in Table 1. In other embodiments, the VH and / or VL amino acid sequences may be identical except for amino acid substitutions at no more than 1, 2, 3, 4, or 5 amino acid positions. Antibodies having VH and VL regions with high (i.e., 80% or greater) identity to those described in Table 1 can be obtained by mutagenesis (e.g., site-directed or PCR-mediated mutagenesis) of nucleic acid molecules encoding SEQ ID NO: 10 or 30 and SEQ ID NO: 20 and 40, respectively, and then testing the encoded altered antibodies for retained function using the functional assays described herein.

[0148] In other embodiments for use in the methods described herein, the full-length heavy chain and / or full-length light chain amino acid sequences may be 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences listed in Table 1 (e.g., SEQ ID NOs: 11 and / or 21, or 31 and / or 41). In other embodiments for use in the formulations described herein, the full-length heavy chain and / or full-length light chain amino acid sequences may be 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences listed in Table 1 (e.g., SEQ ID NOs: 11 and / or 21, or 31 and / or 41). Antibodies having full-length heavy chains and full-length light chains with high (e.g., 80% or higher) identity to the full-length heavy chain of any one of SEQ ID NOs: 11 or 31 and the full-length light chain of any one of SEQ ID NOs: 21 or 41 can be obtained by mutagenesis (e.g., site-directed or PCR-mediated mutagenesis) of nucleic acid molecules encoding such polypeptides, and then testing the retained function of the encoded altered antibodies using the functional assays described herein.

[0149] In one aspect, provided herein is an isolated antibody or a functional antigen-binding fragment thereof, comprising a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 11 and 31; the light chain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 41; and the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey and baboon FXIa). In one embodiment, the isolated antibody or its functional antigen-binding fragment comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 11; the light chain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 21; and the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey and baboon FXIa). In one embodiment, the isolated antibody or its functional antigen-binding fragment comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 31; the light chain comprises an amino acid sequence that is at least 80%, at least 90% or at least 95% identical to the amino acid sequence of SEQ ID NO: 41; and the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey and baboon FXIa). In certain aspects of the present disclosure, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences defined by Kabat, such as SEQ ID NOs: 3, 4, 5, 13, 14 and 15, respectively. In certain other aspects of the present disclosure, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences defined by Chothia, such as SEQ ID NOs: 6, 7, 8, 16, 17 (or the amino acid sequence of KNY) and 18, respectively. In certain other aspects, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences defined by Combinatorial Systems, such as SEQ ID NOs: 46, 4, 5, 33, 14 and 15, respectively. In certain other aspects, the heavy and light chain sequences further comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences defined by IMGT, such as SEQ ID NOs: 43, 44, 45, 47, 37 (or the amino acid sequence of KNY) and 15, respectively.

[0150] In other embodiments for use in the methods described herein, the full-length heavy chain and / or full-length light chain nucleotide sequences may be 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences listed in Table 1 (e.g., SEQ ID NOs: 12 and / or 22, or 32 and / or 42).

[0151] In other embodiments for use in the methods described herein, the heavy chain variable region and / or light chain variable region nucleotide sequences may be 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences listed in Table 1 (e.g., SEQ ID NOs: 10 and / or 20, or 30 and / or 40). In other embodiments for use in the formulations described herein, the heavy chain variable region and / or light chain variable region nucleotide sequences may be 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequences listed in Table 1 (e.g., SEQ ID NOs: 10 and / or 20, or 30 and / or 40).

[0152] As used herein, the percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced to achieve optimal alignment of the two sequences, and the length of each gap (i.e., % identity equals number of identical positions / total number of positions x 100). The comparison of sequences between two sequences and determination of percent identity can be accomplished using a mathematical algorithm, as described in the non-limiting examples below.

[0153] As described herein, the isolated anti-FXI and / or FXIa antibodies or antigen-binding fragments thereof may be monoclonal antibodies, human antibodies or humanized antibodies, chimeric antibodies, single-chain antibodies, Fab fragments, Fv fragments, F(ab')2 fragments or scFv fragments, and / or IgG isotypes (e.g., IgG1, such as human IgG1). In specific embodiments, the anti-FXI and / or anti-FXIa antibodies described herein are recombinant human antibodies. In specific embodiments, the anti-FXI and / or anti-FXIa antibodies described herein are human IgG1 / lambda (λ) antibodies. In specific embodiments, the anti-FXI and / or anti-FXIa antibodies described herein are human IgG1 / lambda (λ) antibodies comprising an Fc domain that is engineered to reduce the likelihood of effector function (e.g., ADCC and / or CDC), such as human Fc domains comprising D265A and / or P329A substitutions.

[0154] Additionally or alternatively, the protein sequences disclosed herein can be further used as "query sequences" to search public databases, e.g., to identify related sequences. For example, such searches can be performed using the BLAST program (version 2.0) of Altschul et al., 1990 J. Mol. Biol. 215:403-10.

[0155] Antibodies with conservative modifications

[0156] In certain other embodiments, the antibodies of the present disclosure for use in the methods described herein (e.g., methods for treating a subject having or at risk of having a thromboembolic disorder) have a heavy chain variable region sequence comprising a CDR1, CDR2, and CDR3 sequence and a light chain variable region comprising a CDR1, CDR2, and CDR3 sequence, wherein one or more of these CDR sequences have a specific amino acid sequence based on an antibody described herein or a conservative modification thereof, and wherein the antibody retains the desired functional properties of the FXIa-binding antibodies of the present disclosure. In certain other embodiments, the antibodies of the present disclosure for use in the formulations described herein (e.g., formulations in vials, intravenous drug delivery formulations) have a heavy chain variable region sequence comprising a CDR1, CDR2, and CDR3 sequence and a light chain variable region comprising a CDR1, CDR2, and CDR3 sequence, wherein one or more of these CDR sequences have a specific amino acid sequence based on an antibody described herein or a conservative modification thereof, and wherein the antibody retains the desired functional properties of the FXIa-binding antibodies of the present disclosure.

[0157] Thus, for use in the methods described herein, in some embodiments, the present disclosure provides an isolated antibody or antigen-binding fragment thereof consisting of a heavy chain variable region comprising CDR1, CDR2 and CDR3 sequences and a light chain variable region comprising CDR1, CDR2 and CDR3 sequences, wherein: the heavy chain variable region CDR1 amino acid sequence is selected from the group consisting of SEQ ID NOs: 3 and 23 and conservative modifications thereof; the heavy chain variable region CDR2 amino acid sequence is selected from the group consisting of SEQ ID NOs: 4 and 24 and conservative modifications thereof; the heavy chain variable region CDR3 amino acid sequence is selected from the group consisting of SEQ ID NOs: 5 and 25 and conservative modifications thereof; the light chain variable region CDR1 amino acid sequence is selected from the group consisting of SEQ ID NOs: 13 and 33 and conservative modifications thereof; the light chain variable region CDR2 amino acid sequence is selected from the group consisting of SEQ ID NOs: 14 and 34 and conservative modifications thereof; the light chain variable region of CDR3 amino acid sequence is selected from the group consisting of SEQ ID NOs: 15 and 35 and conservative modifications thereof; and the antibody or antigen-binding fragment thereof specifically binds to FXIa.

[0158] For use in the formulations described herein, in some embodiments, the present disclosure provides an isolated antibody or antigen-binding fragment thereof, which consists of a heavy chain variable region comprising CDR1, CDR2 and CDR3 sequences and a light chain variable region comprising CDR1, CDR2 and CDR3 sequences, wherein: the heavy chain variable region CDR1 amino acid sequence is selected from the group consisting of SEQ ID NOs: 3 and 23 and conservative modifications thereof; the heavy chain variable region CDR2 amino acid sequence is selected from the group consisting of SEQ ID NOs: 4 and 24 and conservative modifications thereof; the heavy chain variable region CDR3 amino acid sequence is selected from the group consisting of SEQ ID NOs: 5 and 25 and conservative modifications thereof; the light chain variable region CDR1 amino acid sequence is selected from the group consisting of SEQ ID NOs: 13 and 33 and conservative modifications thereof; the light chain variable region CDR2 amino acid sequence is selected from the group consisting of SEQ ID NOs: 14 and 34 and conservative modifications thereof; the light chain variable region of CDR3 amino acid sequence is selected from the group consisting of SEQ ID NOs: 15 and 35 and conservative modifications thereof; and the antibody or antigen-binding fragment thereof specifically binds to FXIa.

[0159] In one aspect, the present invention provides an isolated antibody or an antigen-binding fragment thereof, which consists of a heavy chain variable region comprising CDR1, CDR2 and CDR3 sequences and a light chain variable region comprising CDR1, CDR2 and CDR3 sequences, wherein: the heavy chain variable region CDR1 amino acid sequence is selected from the group consisting of those shown in Table 1 and conservative modifications thereof; the heavy chain variable region CDR2 amino acid sequence is selected from the group consisting of those shown in Table 1 and conservative modifications thereof; the heavy chain variable region CDR3 amino acid sequence is selected from the group consisting of those shown in Table 1 and conservative modifications thereof; the light chain variable region CDR1 amino acid sequence is selected from the group consisting of those shown in Table 1 and conservative modifications thereof; the light chain variable region CDR2 amino acid sequence is selected from the group consisting of those shown in Table 1 and conservative modifications thereof; the light chain variable region of CDR3 amino acid sequence is selected from the group consisting of those shown in Table 1 and conservative modifications thereof; and the antibody or antigen-binding fragment thereof specifically binds to FXIa.

[0160] In other embodiments for use in the methods described herein, the antibodies of the present disclosure are optimized for expression in mammalian cells, have a full-length heavy chain sequence and a full-length light chain sequence, wherein one or more of these sequences have a specified amino acid sequence based on an antibody described herein or a conservative modification thereof, and wherein the antibody retains the desired functional properties of the FXIa-binding antibodies of the present disclosure. In other embodiments for use in the formulations described herein, the antibodies of the present disclosure are optimized for expression in mammalian cells, have a full-length heavy chain sequence and a full-length light chain sequence, wherein one or more of these sequences have a specified amino acid sequence based on an antibody described herein or a conservative modification thereof, and wherein the antibody retains the desired functional properties of the FXIa-binding antibodies of the present disclosure. Thus, the present disclosure provides an isolated antibody optimized for expression in mammalian cells, consisting of a full-length heavy chain and a full-length light chain, wherein the full-length heavy chain has an amino acid sequence selected from the group consisting of SEQ ID NO: 11 or 31 and conservative modifications thereof; and the full-length light chain has an amino acid sequence selected from the group consisting of SEQ ID NO: 21 or 41 and conservative modifications thereof; and the antibody specifically binds to FXI and / or FXIa (e.g., human, rabbit, cynomolgus monkey, and baboon FXIa).

[0161] Antibodies that bind to the same epitope

[0162] In some embodiments, the present disclosure provides antibodies that compete for the same epitope as the FXI and / or FXIa binding antibodies described in Table 1 for use in the methods described herein (e.g., methods for treating a subject having or at risk of having a thromboembolic disorder). In some embodiments, the present disclosure provides antibodies that compete for the same epitope as the FXI and / or FXIa binding antibodies described in Table 1 for use in the formulations described herein (e.g., formulations in vials, intravenous drug delivery formulations). Thus, additional antibodies can be identified based on their ability to compete (e.g., competitively inhibit binding in a statistically significant manner by binding to the same or overlapping epitope) with other antibodies of the present disclosure in a FXI and / or FXIa binding assay (e.g., those described in the Examples section). The ability of a test antibody to inhibit binding of an antibody of the present disclosure to a FXI and / or FXIa protein indicates that the test antibody can compete with the antibody for binding to FXI and / or FXIa; according to non-limiting theory, such an antibody can bind to the same or related (e.g., structurally similar or spatially proximal) epitope on the FXI and / or FXIa protein as the antibody it competes with. In a certain embodiment, the antibody that binds to the same epitope on FXI and / or FXIa as the antibodies of the present disclosure is a human monoclonal antibody. Such human monoclonal antibodies can be prepared and isolated as described herein.

[0163] As used herein, an antibody "competes" for binding when the competing antibody binds to the same FXI and / or FXIa epitope as an antibody or antigen-binding fragment of the present disclosure (e.g., Antibody 1 or Antibody 2) and inhibits FXI and / or FXIa binding of the antibody or antigen-binding fragment of the present disclosure by more than 50% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) in the presence of an equimolar concentration of the competing antibody. This can be determined, for example, in a competitive binding assay by any method well known to those skilled in the art.

[0164] As used herein, an antibody or antigen-binding fragment thereof does not "compete" with an FXI and / or FXIa antibody or antigen-binding fragment of the present disclosure (e.g., Antibody 1 or Antibody 2) unless the competing antibody or antigen-binding fragment thereof binds to the same FXI and / or FXIa epitope as the antibody or antigen-binding fragment of the present disclosure, or to an overlapping FXI and / or FXIa epitope. As used herein, a competing antibody or antigen-binding fragment thereof does not include one that (i) sterically blocks an antibody or antigen-binding fragment of the present disclosure from binding to its target (e.g., if the competing antibody binds to a nearby non-overlapping FXI and / or FXIa epitope and physically prevents the antibody or antigen-binding fragment of the present disclosure from binding to its target); and / or (ii) binds to a different non-overlapping FXI and / or FXIa epitope and induces a conformational change in the FXI and / or FXIa protein such that the protein is no longer able to be bound by an FXI and / or FXIa antibody or antigen-binding fragment of the present disclosure in the manner that would occur in the absence of the conformational change.

[0165] Engineered and modified antibodies

[0166] In some embodiments, the antibodies of the present disclosure used in the methods described herein can be further prepared using antibodies having one or more VH and / or VL sequences shown herein as starting materials to engineer modified antibodies, which can have different properties from the starting antibodies. In some embodiments, the antibodies of the present disclosure used in the formulations described herein can be further prepared using antibodies having one or more VH and / or VL sequences shown herein as starting materials to engineer modified antibodies, which can have different properties from the starting antibodies. Antibodies can be engineered by modifying one or more residues within one or two variable regions (i.e., VH and / or VL), such as within one or more CDR regions and / or within one or more framework regions. Additionally or alternatively, antibodies can be engineered by modifying residues within the constant region, for example, to change the effector function of the antibody.

[0167] One type of variable region engineering that can be performed is CDR transplantation. Antibodies interact with target antigens primarily via amino acid residues located in the six heavy and light chain complementary determining regions (CDRs). For this reason, the amino acid sequences within the CDRs are more diverse between individual antibodies than sequences outside the CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, it is possible to express recombinant antibodies that mimic the properties of a specific naturally occurring antibody by constructing an expression vector that includes the CDR sequences from a specific naturally occurring antibody grafted onto framework sequences from a different antibody with different properties (see, e.g., Riechmann, L. et al., 1998 Nature 332:323-327; Jones, P. et al., 1986 Nature 321:522-525; Queen, C. et al., 1989 Proc. Natl. Acad., USA 86:10029-10033; U.S. Pat. No. 5,225,539 to Winter, and U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.).

[0168] Therefore, another embodiment of the present disclosure relates to an isolated antibody or antigen-binding fragment thereof, comprising a heavy chain variable region comprising a CDR1 sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 3 and 23, respectively; a CDR2 sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 and 24; a CDR3 sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 5 and 25; and a light chain variable region having a CDR1 sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 13 and 33, respectively; a CDR2 sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 14 and 34; and a CDR3 sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 15 and 35. Thus, such antibodies contain the VH and VL CDR sequences of monoclonal antibodies, but may contain framework sequences different from those of these antibodies.

[0169] Such framework sequences can be obtained from public DNA databases or published references including germline antibody gene sequences. For example, germline DNA sequences of human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database and Kabat, EA et al., 1991 Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242; Tomlinson, IM et al., 1992 J. Mol. Biol. 227:776-798; and Cox, JPL et al., 1994 Eur. J Immunol. 24:827-836; the contents of each of which are expressly incorporated herein by reference.

[0170] Examples of framework sequences for antibodies of the present disclosure are those that are structurally similar to the framework sequences used by the selected antibodies of the present disclosure, for example, the consensus sequences and / or framework sequences used by the monoclonal antibodies of the present disclosure. The VH CDR1, 2 and 3 sequences and the VL CDR1, 2 and 3 sequences can be transplanted to a framework region having a sequence identical to that found in the germline immunoglobulin gene from which the framework sequence is derived, or the CDR sequence can be transplanted to a framework region containing one or more mutations compared to the germline sequence. For example, it has been found that in some cases, it is beneficial to mutate residues within the framework region to maintain or enhance the antigen binding ability of the antibody (see, for example, U.S. Pat. Nos. 5,530,101 to Queen et al.; 5,585,089; 5,693,762 and 6,180,370). Frameworks that can be used as scaffolds on which the antibodies and antigen-binding fragments described herein are constructed include, but are not limited to, VH1A, VH1B, VH3, Vk1, Vl2 and Vk2.

[0171] Accordingly, for use in the methods described herein, another embodiment of the present disclosure relates to an isolated FXIa-binding antibody or antigen-binding fragment thereof, comprising a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 29, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions in the framework regions of such sequences, and further comprising a light chain variable region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 19 and 39, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions in the framework regions of such sequences.

[0172] Accordingly, for use in the formulations described herein, another embodiment of the present disclosure relates to an isolated FXIa-binding antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 29, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions in the framework regions of such sequences, and further comprising a light chain variable region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 19 and 39, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions in the framework regions of such sequences.

[0173] Another type of variable region modification is the mutation of amino acid residues within the VH and / or VL CDR1, CDR2 and / or CDR3 regions to improve one or more binding properties (e.g., affinity) of the antibody of interest, referred to as "affinity" maturation. Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce mutations, and the effects on antibody binding or other functional properties of interest can be evaluated in in vitro or in vivo assays as described herein and provided in the Examples section. Conservative modifications (as discussed above) can be introduced. Mutations can be amino acid substitutions, additions or deletions. In addition, typically no more than one, two, three, four or five residues are altered within the CDR regions.

[0174] Thus, in another embodiment for use in the methods described herein, the present disclosure provides an isolated FXIa-binding antibody or antigen-binding fragment thereof consisting of: a heavy chain variable region having a VH CDR1 region consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 3 and 23, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 3 and 23; a VH CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 and 24, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 4 and 24; a VH CDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 5 and 25, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 5 and 25; a VL CDR1 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 13 and 33, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: A VL CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 14 and 34, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 14 and 34; and a VL CDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 15 and 35, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 15 and 35.

[0175] Thus, in another embodiment for use in the methods described herein, the present disclosure provides an isolated FXIa-binding antibody or antigen-binding fragment thereof consisting of: a heavy chain variable region having a VH CDR1 region consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 6 and 26, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 6 and 26; a VH CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 7 and 27, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 7 and 27; a VH CDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 8 and 28, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 8 and 28; a VL CDR1 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 16 and 36, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 16 and 36; a VL CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 and 37 or an amino acid sequence having KNY, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 17 and 37, or an amino acid sequence having KNY; and a VLCDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 18 and 38 or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 18 and 38.

[0176] Thus, in another embodiment for use in the formulations described herein, the present disclosure provides an isolated FXIa-binding antibody or antigen-binding fragment thereof consisting of: a heavy chain variable region having a VH CDR1 region consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 3 and 23, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 3 and 23; a VH CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 and 24, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 4 and 24; a VH CDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 5 and 25, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 5 and 25; a VL CDR1 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 13 and 33, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: A VL CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 14 and 34, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 14 and 34; and a VL CDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 15 and 35, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 15 and 35.

[0177] Thus, in another embodiment for use in the formulations described herein, the present disclosure provides an isolated FXIa-binding antibody or antigen-binding fragment thereof consisting of: a heavy chain variable region having a VH CDR1 region consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 6 and 26, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 6 and 26; a VH CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 7 and 27, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 7 and 27; a VH CDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 8 and 28, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 8 and 28; a VL CDR1 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 16 and 36, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: A VL CDR2 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 and 37 or an amino acid sequence of KNY, or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 17 and 37 (or the amino acid sequence of KNY); and a VLCDR3 region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 18 and 38 or an amino acid sequence having one, two, three, four or five amino acid substitutions, deletions or additions compared to SEQ ID NOs: 18 and 38.

[0178] Antibodies with extended half-life

[0179] In some embodiments, the present disclosure provides antibodies that specifically bind to FXIa protein and have a prolonged half-life in vivo for use in the methods or formulations described herein.

[0180] Many factors can affect the in vivo half-life of a protein. For example, renal filtration, liver metabolism, proteolytic enzyme (protease) degradation, and immunogenic reactions (e.g., neutralization of proteins by antibodies and uptake by macrophages and dendritic cells). A variety of strategies can be used to extend the half-life of the disclosed antibodies. For example, by chemical connection with polyethylene glycol (PEG), reCODE PEG, antibody scaffolds, polysialic acid (PSA), hydroxyethyl starch (HES), albumin binding ligands, and carbohydrate shields; by genetic fusion with proteins such as albumin, IgG, FcRn that bind to serum proteins, and transfer; by (genetic or chemical) coupling to other binding moieties that bind to serum proteins, such as nanobodies, Fabs, DARPins, avimers, affibodies, and anticalins; by genetic fusion with rPEG, albumin, albumin domains, albumin binding proteins, and Fc; or by incorporation into nanocarriers, sustained-release formulations, or medical devices.

[0181] In order to prolong the serum circulation of antibodies in vivo, inert polymer molecules such as high molecular weight PEG can be conjugated to the N-terminus or C-terminus of the antibody via PEG or the site-specific conjugation of the ε-amino group present on the lysine residue, with or without a multifunctional linker, to the antibody or its fragment. In order to PEGylate the antibody, the antibody or its fragment is typically reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions where one or more PEG groups are connected to the antibody or antibody fragment. Pegylation can be carried out by acylation or alkylation with a reactive PEG molecule (or a similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any form of PEG that has been used to derivatize other proteins, such as mono (C1-C10) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody to be PEGylated is a non-glycosylated antibody. Linear or branched polymer derivatization resulting in minimal loss of biological activity will be used. The degree of conjugation can be closely monitored by SDS-PAGE and mass spectrometry to ensure that the PEG molecule is properly conjugated to the antibody. Unreacted PEG can be separated from the antibody-PEG conjugate by size exclusion or by ion exchange chromatography. The binding activity and in vivo efficacy of PEG-derived antibodies can be tested using methods well known to those skilled in the art, such as by immunoassays as described herein. The method of protein PEGylation is known in the art and can be applied to the antibodies of the present disclosure. See, for example, EP 0 154 316 of Nishimura et al. and EP 0 401 384 of Ishikawa et al.

[0182] Other improved PEGylation technologies include Reconstructive Chemical Orthogonal Directed Engineering (ReCODE PEG), which incorporates chemically specified side chains into biosynthetic proteins via a reconstructive system including tRNA synthetase and tRNA. This technology can incorporate more than 30 new amino acids into biosynthetic proteins in E. coli, yeast, and mammalian cells. tRNA incorporates unnatural amino acids at any position where an amber codon is located, converting amber from a stop codon to a codon that signals the incorporation of a chemically specific amino acid.

[0183] Recombinant PEGylation technology (rPEG) can also be used to extend serum half-life. This technology involves genetically fusing an unstructured protein tail of 300-600 amino acids into an existing pharmaceutical protein. Since the apparent molecular weight of such unstructured protein chains is about 15 times greater than their actual molecular weight, the serum half-life of the protein is greatly increased. Compared with traditional PEGylation that requires chemical conjugation and repurification, the manufacturing process is greatly simplified and the product is homogeneous.

[0184] Polysialylation is another technology that uses the natural polymer polysialic acid (PSA) to extend the active life and improve the stability of therapeutic peptides and proteins. PSA is a polymer of sialic acid (a sugar). When used for protein and therapeutic peptide drug delivery, polysialic acid provides a protective microenvironment for conjugation. This increases the active life of therapeutic proteins in circulation and prevents them from being recognized by the immune system. PSA polymers occur naturally in the human body. It is adopted by certain bacteria, which have evolved over millions of years to use it to cover their walls. These naturally polysialylated bacteria are then able to defeat the body's defense system by molecular mimicry. PSA is nature's ultimate stealth technology, which can be easily mass-produced from such bacteria and has predetermined physical characteristics. Bacterial PSA is completely non-immunogenic, even when coupled to proteins, because it is chemically identical to PSA in the human body.

[0185] Another technique involves the use of hydroxyethyl starch ("HES") derivatives linked to antibodies. HES is a modified natural polymer derived from waxy corn starch and can be metabolized by human enzymes. HES solutions are typically administered to replace insufficient blood volume and improve the rheological properties of blood. HESylation of antibodies can increase biological activity by increasing the stability of the molecule and by prolonging the circulation half-life by reducing renal clearance. By varying different parameters, such as the molecular weight of HES, a wide range of HES antibody conjugates can be customized.

[0186] Antibodies with increased in vivo half-life can also be generated by introducing one or more amino acid modifications (i.e., substitutions, insertions, or deletions) into the IgG constant domain or its FcRn binding fragment (preferably Fc or hinge Fc domain fragment). See, e.g., International Publication No. WO 98 / 23289; International Publication No. WO 97 / 34631; and U.S. Pat. No. 6,277,375.

[0187] In addition, the antibody can be conjugated to albumin (e.g., human serum albumin; HSA) to make the antibody or antibody fragment more stable in vivo or have a longer half-life in vivo. Such techniques are well known in the art, see, for example, International Publication Nos. WO 93 / 15199, WO 93 / 15200, and WO 01 / 77137; and European Patent No. EP 413,622. In addition, in the case of bispecific antibodies as described above, the specificity of the antibody can be designed so that one binding domain of the antibody binds to FXIa and the second binding domain of the antibody binds to serum albumin, preferably HSA.

[0188] The strategy of increasing half-life is particularly useful in nanobodies, fibronectin-based binders, and other antibodies or proteins where increased in vivo half-life is desired.

[0189] Antibody conjugates

[0190] In some embodiments, the present disclosure provides antibodies or fragments thereof for use in the methods or formulations described herein, which specifically bind to a FXIa protein recombinantly fused or chemically conjugated (including covalent and non-covalent conjugations) to a heterologous protein or polypeptide (or a fragment thereof, preferably a polypeptide of at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90 or at least 100 amino acids) to produce a fusion protein. In particular, the present disclosure provides a fusion protein comprising an antigen-binding fragment of an antibody described herein (e.g., a Fab fragment, a Fd fragment, a Fv fragment, a F(ab)2 fragment, a VH domain, a VH CDR, a VL domain or a VL CDR) and a heterologous protein, polypeptide or peptide. Methods of fusing or conjugating a protein, polypeptide or peptide to an antibody or antibody fragment are known in the art. See, e.g., U.S. Pat. Nos. 5,336,603, 5,622,929, 5,359,046, 5,349,053, 5,447,851, and 5,112,946; European Patent Nos. EP 307,434 and EP 367,166; International Publication Nos. WO 96 / 04388 and WO 91 / 06570; Ashkenazi et al., 1991, Proc. Natl. Acad. Sci. USA 88: 10535-10539; Zheng et al., 1995, J. Immunol. 154:5590-5600; and Vil et al., 1992, Proc. Natl. Acad. Sci. USA 89: 11337- 11341.

[0191] Additional fusion proteins can be generated via gene shuffling, motif shuffling, exon shuffling and / or codon shuffling (collectively referred to as "DNA shuffling") techniques. DNA shuffling can be used to alter the activity of an antibody or fragment thereof of the present disclosure (e.g., an antibody or fragment thereof with higher affinity and lower dissociation rate). See generally U.S. Pat. Nos. 5,605,793, 5,811,238, 5,830,721, 5,834,252, and 5,837,458; Patten et al., 1997, Curr. Opinion Biotechnol. 8:724-33; Harayama, 1998, Trends Biotechnol. 16(2):76-82; Hansson et al., 1999, J. Mol. Biol. 287:265-76; and Lorenzo and Blasco, 1998, Biotechniques 24(2):308-313 (each of these patents and publications is hereby incorporated by reference in its entirety). Antibodies or fragments thereof, or encoded antibodies or fragments thereof, may be altered by random mutagenesis prior to recombination by error-prone PCR, random nucleotide insertion, or other methods. The polynucleotide encoding the antibody or fragment thereof that specifically binds to FXIa protein can be recombined with one or more components, motifs, segments, parts, domains, fragments, etc. of one or more heterologous molecules.

[0192] In addition, the antibody or its fragment can be fused with a marker sequence such as a peptide to facilitate purification. In certain embodiments, the marker amino acid sequence is a hexa-histidine peptide (amino acid sequence: HHHHHH; SEQ ID NO: 48), such as a tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, CA, 91311), many of which are commercially available. As described in Gentz ​​et al., 1989, Proc. Natl. Acad. Sci. USA 86: 821-824, for example, hexa-histidine (SEQ ID NO: 48) provides convenient purification of fusion proteins. Other peptide tags that can be used for purification include, but are not limited to, hemagglutinin ("HA") tags, which correspond to epitopes derived from influenza hemagglutinin proteins (Wilson et al., 1984, Cell 37: 767), and "flag" tags.

[0193] In other embodiments, the antibodies or fragments thereof disclosed herein are conjugated to a diagnostic agent or a detectable agent. Such antibodies can be used to monitor or predict the occurrence, development, progression and / or severity of a disease or condition as part of a clinical testing procedure, such as to determine the efficacy of a particular therapy. Such diagnosis and detection can be accomplished by coupling the antibody to a detectable substance, which includes, but is not limited to, various enzymes, such as, but not limited to, horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; prosthetic groups, such as, but not limited to, streptavidin-biotin and avidin / biotin; fluorescent materials, such as, but not limited to, umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazineamine fluorescein, dansyl chloride, or phycoerythrin; luminescent materials, such as, but not limited to, luminol; bioluminescent materials, such as, but not limited to, luciferase, luciferin, and aequorin; radioactive materials , such as but not limited to iodine (131I, 125I, 123I and 121I), carbon (14C), sulfur (35S), tritium (3H), indium (115In, 113In, 112In and 111In), technetium (99Tc), thallium (201Ti), gallium (68Ga, 67Ga), palladium (103Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), 153Sm, 177Lu, 159Gd, 149Pm, 140La, 175Yb, 166Ho, 90Y, 47Sc, 186Re, 188Re, 142 Pr, 105Rh, 97Ru, 68Ge, 57Co, 65Zn, 85Sr, 32P, 153Gd, 169Yb, 51Cr, 54Mn, 75Se, 113Sn and 117Tin; and positron emitting metals using various positron emission tomography, and non-radioactive paramagnetic metal ions.

[0194] In some embodiments, the present disclosure also encompasses the use of antibodies or fragments thereof conjugated to a therapeutic moiety. The antibody or fragment thereof may be conjugated to a therapeutic moiety, such as a cytotoxin, e.g., a cytostatic or cytocidal agent, a therapeutic agent, or a radioactive metal ion, e.g., an alpha-emitter. Cytotoxins or cytotoxic agents include any agent that is harmful to cells.

[0195] In addition, the antibody or its fragment can be conjugated to a therapeutic or drug moiety that modifies a given biological response. The therapeutic or drug moiety should not be construed as being limited to classical chemotherapeutic agents. For example, the drug moiety can be a protein, peptide, or polypeptide having a desired biological activity. Such proteins can include, for example, toxins such as abrin, ricin A, Pseudomonas exotoxin, cholera toxin, or diphtheria toxin; proteins such as tumor necrosis factor, alpha-interferon, beta-interferon, nerve growth factor, platelet-derived growth factor, tissue plasminogen activator, apoptotic agents, anti-angiogenic agents; or biological response modifiers such as lymphokines.

[0196] In addition, the antibody can be conjugated to a therapeutic moiety, such as a radioactive metal ion, such as an α-emitter, such as 213Bi, or a macrocyclic chelator that can be used to conjugate radioactive metal ions including but not limited to 131In, 131LU, 131Y, 131Ho, 131Sm to a polypeptide. In certain embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), which can be attached to the antibody via a linker molecule. Such linker molecules are well known in the art and are described in Denardo et al., 1998, Clin Cancer Res. 4(10):2483-90; Peterson et al., 1999, Bioconjug. Chem. 10(4):553-7; and Zimmerman et al., 1999, Nucl. Med. Biol. 26(8):943-50, each of which is incorporated by reference in its entirety.

[0197] The techniques for conjugating therapeutic moieties to antibodies are well known, see, e.g., Arnon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery”, Controlled Drug Delivery (2nd ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, Monoclonal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); “Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., 1982, Immunol. Rev. 62: 119-58.

[0198] The antibodies can also be attached to solid supports, which are particularly useful for immunoassays or purification of target antigens. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene.

[0199] Pharmaceutical preparations

[0200] In some embodiments, the present disclosure also provides a pharmaceutical preparation containing a therapeutically effective amount of factor XI and / or factor XIa antibodies (eg, antibody 1) disclosed herein. The pharmaceutical preparation comprises one or more excipients and is maintained at a certain pH. As used herein, non-limiting examples of "excipients" include any non-therapeutic agents added to the preparation to provide the desired physical or chemical properties, such as pH, osmotic pressure, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption or penetration. Exemplary pharmaceutical preparations comprising factor XI and / or factor XIa antibodies (eg, antibody 1) disclosed herein are provided in, for example, International Application Publication WO2021 / 127525.

[0201] Drug substances

[0202] Antibody 1 is a high affinity anti-human factor XI monoclonal antibody. It is expressed in a Chinese hamster ovary cell line (CHO-C8TD). In some embodiments, the Antibody 1 drug substance is fully formulated for subcutaneous administration (i.e., no other excipients are added), and is therefore identical in composition to the Antibody 1 drug product. In some embodiments, for intravenous administration, the Antibody 1 drug product is further diluted in an appropriate carrier. In some embodiments, the Antibody 1 drug product is diluted in a solution comprising dextrose, such as a 5% dextrose in water solution (D5W).

[0203] Excipients and pH

[0204] In some embodiments, the excipients contained in the antibody 1 drug product are pharmacopoeia grade excipients. In some embodiments, the excipients in the antibody 1 drug product include histidine, histidine salts, sugars and polysorbates. In some embodiments, the excipients in the antibody 1 drug product include L-histidine and L-histidine hydrochloride monohydrate (histidine buffer), sucrose and polysorbate 20. Excipients can be selected according to their suitability for intravenous and subcutaneous administration, thereby providing the necessary stability, buffering capacity and tension. The formulation maximizes the stability of the monoclonal antibody product and can provide a sterile solution suitable for subcutaneous or intravenous administration. In some embodiments, sugar (e.g., sucrose) acts as a stabilizer. In some embodiments, histidine (e.g., L-histidine, L-histidine HCl monohydrate) acts as a buffer. In some embodiments, polysorbates (e.g., polysorbate 20) acts as a stabilizer. In some embodiments, the formulation is adjusted to the final volume in water for injection (WFI).

[0205] One or more excipients in the pharmaceutical formulation of the present invention include a buffer. As used herein, the term "buffer" refers to one or more components that, when added to an aqueous solution, can protect the solution from pH changes when an acid or base is added or when diluted with a solvent. In addition to phosphate buffers, glycine, carbonate, citrate, histidine buffers, etc. can also be used, in which case sodium ions, potassium ions, or ammonium ions can be used as counter ions.

[0206] In certain embodiments, the buffer or buffer system comprises at least one buffer having a buffer range that completely or partially overlaps with the range of pH 5.0-7.4. In certain embodiments, the buffer has a pH of about 5.5 ± 0.5. In certain embodiments, the buffer comprises a histidine buffer. In certain embodiments, the histidine buffer is 0.05-10 mM, 0.1-10 mM, 0.2-10 mM, 0.5-10 mM, 1-10 mM, 5-10 mM, 5 to 100 mM, 10 to 100 mM, 15 to 100 mM, 20 to 100 mM, 30 to 100 mM, 40 to 100 mM, 50 to 100 mM, 60 to 100 mM, 70 to 100 mM, 80 to 100 mM, 90 to 100 mM, 5 to 90 mM, 5 to 80 mM, 5 to 70 mM, 5 to 60 mM, 5 to 50 mM, 5 to 40 mM, 5 to 30 mM, 5 to 20 mM, 10 to 50 mM, 10 to 40 mM, 10 to 30 In some embodiments, the histidine buffer has a pH of about 5.0, about 5.5, about 6.0, about 6.5 or about 7.0. In a specific embodiment, the histidine buffer has a pH of about 5.5.

[0207] The pharmaceutical preparation of the present invention may have a pH of 5.0 to 6.0. For example, in certain embodiments, the pharmaceutical preparation has a pH of 5.0 to 6.0 (i.e. 5.5 ± 0.5), 5.1 to 5.9 (i.e. 5.5 ± 0.4), 5.2 to 5.8 (i.e. 5.5 ± 0.3), 5.3 to 5.7 (i.e. 5.5 ± 0.2), 5.4 to 5.6 (i.e. 5.5 ± 0.1) or 5.45 to 5.55 (i.e. 5.5 ± 0.05). In certain embodiments, the pharmaceutical preparation has a pH of about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4 or about 6.5. In certain embodiments, the pharmaceutical preparation has a pH of about 5.5. According to the rules of scientific rounding, a pH greater than or equal to 5.45 and less than or equal to 5.55 is rounded to 5.5.

[0208] In certain embodiments, the buffer system of the pharmaceutical formulation comprises 10 to 30 mM histidine at a pH of 5.5 ± 0.2. In certain embodiments, the buffer system of the pharmaceutical formulation comprises about 20 mM histidine at a pH of 5.5 ± 0.2. In certain embodiments, the buffer system of the pharmaceutical formulation comprises 10 to 30 mM histidine at a pH of 5.5 ± 0.05. In certain embodiments, the buffer system of the pharmaceutical formulation comprises about 20 mM histidine at a pH of 5.5 ± 0.05.

[0209] In certain embodiments, the buffer system of the pharmaceutical formulation comprises 0.10 to 0.30 mM histidine at a pH of 5.5 ± 0.2. In certain embodiments, the buffer system of the pharmaceutical formulation comprises about 0.20 mM histidine at a pH of 5.5 ± 0.2. In certain embodiments, the buffer system of the pharmaceutical formulation comprises 0.10 to 0.30 mM histidine at a pH of 5.5 ± 0.05. In certain embodiments, the buffer system of the pharmaceutical formulation comprises about 0.20 mM histidine at a pH of 5.5 ± 0.05.

[0210] One or more excipients in the pharmaceutical preparation of the present invention also include sugar or sugar alcohol.Sugar and sugar alcohol are suitable for use as heat stabilizers in pharmaceutical preparations.In certain embodiments, the pharmaceutical preparation includes sugar, such as monosaccharide (glucose, xylose or erythritol), disaccharide (such as sucrose, trehalose, maltose or galactose) or oligosaccharide (such as stachyose).In specific embodiments, the pharmaceutical preparation includes sucrose.In certain embodiments, pharmaceutical composition includes sugar alcohol, such as sugar alcohol (such as mannitol, sorbitol or xylitol) derived from monosaccharide, sugar alcohol (such as lactitol or maltitol) derived from disaccharide or sugar alcohol derived from oligosaccharide.In specific embodiments, the pharmaceutical preparation includes sucrose.

[0211] The amount of sugar or sugar alcohol contained in the preparation can change according to the specific circumstances and the intended purpose of using the preparation. In certain embodiments, the pharmaceutical preparation includes 50 to 300 mM, 50 to 250 mM, 100 to 300 mM, 100 to 250 mM, 150 to 300 mM, 150 to 250 mM, 200 to 300 mM, 200 to 250 mM or 250 to 300 mM sugar or sugar alcohol. In certain embodiments, the pharmaceutical preparation includes about 50 mM, about 75 mM, about 100 mM, about 125 mM, about 150 mM, about 200 mM, about 220 mM, about 250 mM or about 300 mM sugar or sugar alcohol. In a specific embodiment, the pharmaceutical preparation includes about 220 mM sugar or sugar alcohol (such as sucrose).

[0212] The amount of sugar or sugar alcohol contained in the preparation can vary according to the specific circumstances and the intended purpose of using the preparation. In certain embodiments, the pharmaceutical preparation comprises 0.50 to 3.00 mM, 0.50 to 2.50 mM, 1.00 to 3.00 mM, 1.00 to 2.50 mM, 1.50 to 3.00 mM, 1.50 to 2.50 mM, 2.00 to 3.00 mM, 2.00 to 2.50 mM or 2.50 to 3.00 mM sugar or sugar alcohol. In certain embodiments, the pharmaceutical preparation comprises about 0.50 mM, about 0.75 mM, about 1.00 mM, about 1.25 mM, about 1.50 mM, about 2.00 mM, about 2.20 mM, about 2.50 mM or about 3.00 mM sugar or sugar alcohol. In certain embodiments, the pharmaceutical formulation comprises about 2.20 mM of a sugar or sugar alcohol (eg, sucrose).

[0213] One or more excipients in the pharmaceutical formulation disclosed herein also include a surfactant. As used herein, the term "surfactant" refers to a surface-active molecule containing a hydrophobic portion (e.g., an alkyl chain) and a hydrophilic portion (e.g., a carboxyl group and a carboxylate group). Surfactants can be used in pharmaceutical formulations for reducing aggregation of therapeutic proteins. Surfactants suitable for pharmaceutical formulations are generally nonionic surfactants, and include but are not limited to polysorbates (e.g., polysorbate 20 or 80); Poloxamers (e.g., poloxamer 188); sorbitan and derivatives; Triton; sodium lauryl sulfate; sodium octyl glucoside; lauryl-, myristyl-, linoleyl-, or stearoyl-sulfobetaine; lauryl-, myristyl-, linoleyl-, or stearoyl-sarcosine Acid; linoleyl-, myristyl- or cetyl-betaine; lauroyl-cocamidopropyl-, linoleyl-, myristyl-, palmitamidopropyl- or isostearamidopropyl betaine (e.g., lauroyl); myristyl-, palmitamidopropyl- or isostearamidopropyl-dimethylamine; sodium methyl cocoyl taurate or disodium methyl oleyl taurate; and MONAQUAT™ series (Mona Industries, Inc., Paterson, NJ), polyethylene glycol, polypropylene glycol and copolymers of ethylene and propylene glycol (e.g., Pluronics, PF68, etc.). In certain embodiments, the surfactant is polysorbate. In certain embodiments, the surfactant is polysorbate 20.

[0214] The amount of nonionic surfactant contained in the pharmaceutical preparation of the present invention can change according to the specific characteristics required for the preparation and the specific environment and purpose of the preparation expected use. In certain embodiments, the pharmaceutical preparation includes 0.02% to 0.06%, 0.03% to 0.05% or 0.035% to 0.045% nonionic surfactant (e.g. polysorbate 20). In certain embodiments, the pharmaceutical preparation includes about 0.005%, about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09% or about 0.1% nonionic surfactant (e.g. polysorbate 20).

[0215] The amount of nonionic surfactant contained in the pharmaceutical preparation of the present invention can change according to the specific characteristics required for the preparation and the specific environment and purpose of the preparation expected use. In certain embodiments, the pharmaceutical preparation includes 0.0002% to 0.0006%, 0.0003% to 0.0005% or 0.00035% to 0.00045% nonionic surfactant (e.g. polysorbate 20). In certain embodiments, the pharmaceutical preparation includes about 0.00005%, about 0.0001%, about 0.0002%, about 0.0003%, about 0.0004%, about 0.0005%, about 0.0006%, about 0.0007%, about 0.0008%, about 0.0009% or about 0.001% nonionic surfactant (e.g. polysorbate 20).

[0216] In certain embodiments, the drug product is diluted in an aqueous carrier suitable for the route of administration, such as intravenous administration. Exemplary carriers include sterile water for injection (SWFI), bacteriostatic water for injection (BWFI), pH buffered solutions (e.g., phosphate buffered saline), sterile saline solutions, Ringer's solution, or glucose solution. In one embodiment, when the drug formulation is prepared for intravenous administration, the drug formulation can be diluted in 5% glucose solution (D5W).

[0217] Exemplary Formulations

[0218] In certain embodiments, pharmaceutical formulations of the invention comprise a factor XI and / or factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having the amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has the amino acid sequence of SEQ ID NO: 19 or 39, and the VL has the amino acid sequence of SEQ ID NO: 19 or 39), a histidine buffer, a sugar or sugar alcohol (e.g., sucrose) and a polysorbate (e.g., polysorbate 20), at a pH of 5.5 to 6.5.

[0219] In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having the amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has the amino acid sequence of SEQ ID NO: 19 or 39, and the VL has the amino acid sequence of SEQ ID NO: 39), 10 to 30 mM histidine buffer, 200 to 300 mM sugar or sugar alcohol (e.g., sucrose), and 0.02% to 0.06% polysorbate (e.g., polysorbate 20), at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a factor XI and / or factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 9 or 29, and the VL has an amino acid sequence of SEQ ID NO: 19 or 39), about 20 mM histidine buffer, about 220 mM sugar or sugar alcohol (e.g., sucrose), and about 0.04% polysorbate (e.g., polysorbate 20), at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a factor XI and / or factor XIa antibody, about 20 mM histidine buffer, about 220 mM sugar or sugar alcohol (e.g., sucrose), and about 0.04% polysorbate (e.g., polysorbate 20), at a pH of 5.2 to 5.8. In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 39), about 20 mM histidine buffer, about 220 mM sugar or sugar alcohol (e.g., sucrose), and about 0.04% polysorbate (e.g., polysorbate 20), at a pH of 5.45 to 5.55.

[0220] In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having the amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has the amino acid sequence of SEQ ID NO: 19 or 39, and the VL has the amino acid sequence of SEQ ID NO: 39), 0.10 to 0.30 mM histidine buffer, 2.00 to 3.00 mM sugar or sugar alcohol (e.g., sucrose), and 0.0002% to 0.0006% polysorbate (e.g., polysorbate 20), at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL) having an amino acid sequence of SEQ ID NO: 19 or 39), about 0.20 mM histidine buffer, about 2.20 mM sugar or sugar alcohol (e.g., sucrose), and about 0.0004% polysorbate (e.g., polysorbate 20), at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 39), about 0.20 mM histidine buffer, about 2.20 mM sugar or sugar alcohol (e.g., sucrose), and about 0.0004% polysorbate (e.g., polysorbate 20), at a pH of 5.2 to 5.8. In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 39), about 0.20 mM histidine buffer, about 2.20 mM sugar or sugar alcohol (e.g., sucrose), and about 0.0004% polysorbate (e.g., polysorbate 20), at a pH of 5.45 to 5.55.

[0221] In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a factor XI and / or factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 19 or 39), 10 to 30 mM histidine buffer, 200 to 300 mM sucrose, and 0.02% to 0.06% polysorbate 20, at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a factor XI and / or factor XIa antibody, about 20 mM histidine buffer, about 220 mM sucrose, and about 0.04% polysorbate 20, at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 39), about 20 mM histidine buffer, about 220 mM sucrose, and about 0.04% polysorbate 20, at a pH of 5.3 to 5.7. In certain embodiments, the pharmaceutical formulation comprises 100 to 200 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 39), about 20 mM histidine buffer, about 220 mM sucrose, and about 0.04% polysorbate 20, at a pH of 5.45 to 5.55.

[0222] In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 39), 0.10 to 0.30 mM histidine buffer, 2.00 to 3.00 mM sucrose, and 0.0002% to 0.0006% polysorbate 20, at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a factor XI and / or factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 19 or 39), about 0.20 mM histidine buffer, about 2.20 mM sucrose, and about 0.0004% polysorbate 20, at a pH of 5.0 to 6.0. In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a factor XI and / or factor XIa antibody, 20 mM histidine buffer, about 2.20 mM sucrose, and about 0.0004% polysorbate 20, at a pH of 5.3 to 5.7. In certain embodiments, the pharmaceutical formulation comprises 1.00 to 2.00 mg / mL of a Factor XI and / or Factor XIa antibody (e.g., an antibody having a heavy chain variable domain (VH) having an amino acid sequence of SEQ ID NO: 9 or 29 and a light chain variable domain (VL), wherein the VH has an amino acid sequence of SEQ ID NO: 19 or 39, and the VL has an amino acid sequence of SEQ ID NO: 39), about 0.20 mM histidine buffer, about 2.20 mM sucrose, and about 0.0004% polysorbate 20, at a pH of 5.45 to 5.55.

[0223] In an embodiment, the present disclosure provides a pharmaceutical formulation comprising an antibody that binds to a FXI and / or FXIa protein or an antigen-binding fragment thereof, the pharmaceutical formulation being contained in a vial, wherein the formulation comprises an excess fill volume for completely withdrawing a therapeutically effective amount of an anti-FXI and / or anti-FXIa antibody or an antigen-binding fragment thereof. In certain embodiments, the vial contains a pharmaceutical formulation comprising about 150 mg of an antibody that binds to a FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 9 or 29, and a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 19 or 39; a histidine buffer at a concentration of about 20 mM; a sucrose at a concentration of about 220 mM; and a polysorbate-20 at a concentration of about 0.04% (v / v); and the pH of the formulation is about pH 5.5.

[0224] In an embodiment, the present disclosure provides an intravenously delivered pharmaceutical formulation comprising about 1.5 mg of an antibody or antigen-binding fragment thereof that binds to FXI and / or FXIa protein (e.g., human, rabbit, cynomolgus monkey, and baboon FXI and / or FXIa), the antibody having a heavy chain variable domain (VH), the VH having an amino acid sequence of SEQ ID NO: 9 or 29, and a light chain variable domain (VL), the VL having an amino acid sequence of SEQ ID NO: 19 or 39; a histidine buffer at a concentration of about 0.20 mM; sucrose at a concentration of about 2.20 mM; polysorbate-20 at a concentration of about 0.0004% (v / v) and a diluent (e.g., 5% dextrose in water (D5W)); and the pH of the formulation is about pH 5.5.

[0225] Stability of Factor XI and / or Factor XIa Antibodies

[0226] The pharmaceutical formulations of the present invention exhibit a high level of stability. A pharmaceutical formulation is stable when the Factor XI and / or Factor XIa antibody in the formulation retains an acceptable degree of physical properties, chemical structure and / or biological function after storage under specified conditions.

[0227] Exemplary methods for determining the stability of factor XI and / or factor XIa antibodies in pharmaceutical formulations are described in Example 1 of the present disclosure. In addition, the stability of the protein can be assessed by measuring the binding affinity of the factor XI and / or factor XIa antibodies to their targets or the biological activity of the factor XI and / or factor XIa antibodies in certain in vitro assays, such as the aPTT and FXI activity assays described in WO 2016 / 207858.

[0228] The pharmaceutical formulation can be prepared and stored as a liquid formulation. In certain embodiments, the pharmaceutical formulation is a liquid formulation for storage at 2-8°C (eg, 4°C). In certain embodiments, the pharmaceutical formulation is a liquid formulation for storage at 4°C and protected from light.

[0229] Stability studies found that Antibody 1 150 mg / mL Injection Concentrate was compatible with its excipients and primary packaging materials. Antibody 1 150 mg / mL Injection Concentrate is suitable for subcutaneous administration with a disposable syringe without dilution or dilution with a vehicle buffer such as 5% dextrose (D5W). The dose range of the Injection Concentrate using commercially available disposable syringes has been demonstrated to be 0.5 mg / subject to 600 mg / subject. Materials found to be compatible with Antibody 1 include syringes made of polypropylene or polycarbonate, and injection needles made of stainless steel. Compatibility of Antibody 1 Injection Concentrate has been demonstrated with 1 mL syringes at Antibody 1 concentrations of 0.5 mg / mL to 150 mg / mL. Compatibility of Antibody 1 Injection Concentrate has been demonstrated with 3 mL syringes filled to approximately 2 mL at an Antibody 1 concentration of 150 mg / mL, covering a total dose range of 0.5 mg to 150 mg for 1 mL syringes and a dose of approximately 300 mg per injection for 3 mL syringes (filled with approximately 2 mL).

[0230] Dosage form

[0231] Before drug use, if suitable for route of administration, the pharmaceutical preparation can be diluted in an aqueous carrier. For intravenous administration, suitable carriers include sterile water for injection (SWFI), bacteriostatic water for injection (BWFI), pH buffer solution (e.g., phosphate buffered saline), sterile saline solution, Ringer's solution or glucose solution. For example, when the pharmaceutical preparation is prepared for intravenous administration, the pharmaceutical preparation includes 5% glucose solution (D5W). In certain embodiments, the diluted pharmaceutical preparation is isotonic and is suitable for administration by intravenous infusion, such as D5W. In certain embodiments, the preparation is diluted in about 50 mL D5W, 100 mL D5W, 150 mL D5W, 200 mL D5W, 250 mL D5W, 300 mL D5W, 350 mL D5W, 400 mL D5W, 450 mL D5W, 500 mL D5W or 1 L D5W.

[0232] The pharmaceutical formulation comprises a Factor XI and / or Factor XIa antibody at a concentration suitable for storage. In certain embodiments, the pharmaceutical formulation comprises a Factor XI and / or Factor XIa antibody at a concentration of 100-200 mg / mL, 100-190 mg / mL, 100-180 mg / mL, 100-170 mg / mL, 100-160 mg / mL, 110-150 mg / mL, 120-150 mg / mL, 130-150 mg / mL, 140-150 mg / mL, 140-160 mg / mL, 140-170 mg / mL, 140-180 mg / mL, 140-190 mg / mL, 150-190 mg / mL, 150-180 mg / mL, 150-170 mg / mL, or 150-160 mg / mL. In certain embodiments, the pharmaceutical formulation comprises a Factor XI and / or Factor XIa antibody at a concentration of about 10 mg / mL, about 15 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 120 mg / mL, about 125 mg / mL, about 130 mg / mL, about 135 mg / mL, about 140 mg / mL, about 145 mg / mL, about 150 mg / mL, about 155 mg / mL, about 160 mg / mL, about 165 mg / mL, about 170 mg / mL, about 175 mg / mL, about 180 mg / mL, about 185 mg / mL, about 190 mg / mL, about 195 mg / mL, or about 200 mg / mL.

[0233] The pharmaceutical formulation comprises Factor XI and / or Factor XIa antibody in a concentration suitable for storage. In certain embodiments, the pharmaceutical formulation comprises a Factor XI and / or Factor XIa antibody at a concentration of 1.00-2.00 mg / mL, 1.00-1.90 mg / mL, 1.00-1.80 mg / mL, 1.00-1.70 mg / mL, 1.00-1.60 mg / mL, 1.10-1.50 mg / mL, 1.20-1.50 mg / mL, 1.30-1.50 mg / mL, 1.40-1.50 mg / mL, 1.40-1.60 mg / mL, 1.40-1.70 mg / mL, 1.40-1.80 mg / mL, 1.40-1.90 mg / mL, 1.50-1.90 mg / mL, 1.50-1.80 mg / mL, 1.50-1.70 mg / mL, or 1.50-1.60 mg / mL. In certain embodiments, the pharmaceutical formulation comprises a Factor XI and / or Factor XIa antibody at a concentration of about 0.10 mg / mL, about 0.15 mg / mL, about 0.25 mg / mL, about 0.50 mg / mL, about 0.75 mg / mL, about 1.00 mg / mL, about 1.20 mg / mL, about 1.25 mg / mL, about 1.30 mg / mL, about 1.35 mg / mL, about 1.40 mg / mL, about 1.45 mg / mL, about 1.50 mg / mL, about 1.55 mg / mL, about 1.60 mg / mL, about 1.65 mg / mL, about 1.70 mg / mL, about 1.75 mg / mL, about 1.80 mg / mL, about 1.85 mg / mL, about 1.90 mg / mL, about 1.95 mg / mL, or about 2.00 mg / mL.

[0234] In certain embodiments, the pharmaceutical preparation is packaged in a vial (e.g., vial, bag, pen, or syringe). In certain embodiments, the vial comprises an overfill to allow for complete removal of the expected dose. In certain embodiments, the vial comprises an overfill of 5% to 35%, 10% to 30%, 15% to 25%, or 10% to 20%. In a specific embodiment, the vial comprises about 20% overfill.

[0235] In certain embodiments, the formulation may be a liquid formulation. In certain embodiments, the amount of factor XI and / or factor XIa antibody in the container is suitable for administration as a single dose. In certain embodiments, the amount of factor XI and / or factor XIa antibody in the container is suitable for administration as multiple doses. In certain embodiments, the pharmaceutical formulation comprises factor XI and / or factor XIa antibody in an amount of 0.1 to 200 mg. In certain embodiments, the pharmaceutical preparation comprises 1 to 200 mg, 10 to 200 mg, 20 to 200 mg, 50 to 200 mg, 100 to 200 mg, 200 to 200 mg, 500 to 2000 mg, 1000 to 2000 mg, 0.1 to 1000 mg, 1 to 1000 mg, 10 to 1000 mg, 20 to 1000 mg, 50 to 1000 mg, 100 to 1000 mg, 200 to 1000 mg, 500 to 1000 mg, 0.1 to 500 mg, 1 to 500 mg, 10 to 500 mg, 20 to 500 mg, 50 to 500 mg, 100 to 500 mg, 200 to 500 mg, 0.1 to 200 mg, 1 to 200 mg, 10 to 200 mg, 20 to 200 or 0.1 to 1 mg. In certain embodiments, the pharmaceutical formulation comprises Factor XI and / or Factor XIa antibody in an amount of about 0.1 mg, about 0.5 mg, about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1500 mg, or about 2000 mg in a therapeutically effective amount.

[0236] Dosage regimen and therapeutic use

[0237] In another aspect, the present disclosure provides a method for treating a thromboembolic disease, the method comprising administering a Factor XI and / or Factor XIa antibody disclosed herein (eg, Antibody 1) to a subject in need thereof once a month.

[0238] In certain embodiments, the method further comprises administering factor XI and / or factor XIa antibody to the subject in one or more monthly treatment cycles after the initial treatment cycle, e.g., for a period of 3 months, wherein the factor XI and / or factor XIa antibody is administered on day 1, day 31, and day 61. Subsequent treatment cycles (wherein the subject receives administration of factor XI and / or factor XIa antibody once a month) are designed to maintain a certain level of factor XI and / or factor XIa antibody in the subject. In certain embodiments, the subject receives at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 subsequent treatment cycles. In some embodiments, the subject receives treatment for life.

[0239] In some embodiments, the method includes treating a disease or condition in a subject in need thereof, such as a thromboembolic disease, comprising administering a first dose of an anti-FXI / FXIa antibody or an antigen-binding fragment thereof (e.g., Antibody 1), wherein the first dose is administered intravenously, and administering a second dose of an anti-FXI / FXIa antibody or an antigen-binding fragment thereof (e.g., Antibody 1), wherein the second dose is administered subcutaneously. In some embodiments, the method further comprises administering a third dose subcutaneously. In some embodiments, the method further comprises administering a fourth dose subcutaneously. In some embodiments, the method further comprises administering a fifth dose subcutaneously. In some embodiments, the method further comprises administering a sixth dose subcutaneously. In some embodiments, the method further comprises administering a seventh dose subcutaneously. In some embodiments, the method further comprises administering an eighth dose subcutaneously. In some embodiments, the method further comprises administering a ninth dose subcutaneously. In some embodiments, the method further comprises administering a tenth dose subcutaneously. In some embodiments, the method further comprises administering an eleventh dose subcutaneously. In certain embodiments, the method comprises administering a first dose intravenously, and administering five subsequent doses subcutaneously. In certain embodiments, the duration of treatment is about six months. In certain embodiments, the method comprises administering a first dose intravenously, and eleven subsequent doses subcutaneously. In certain embodiments, the duration of treatment is about one year. In certain embodiments, the method comprises administering a first dose intravenously, and then administering subcutaneous doses monthly until the subject's disease or condition subsides, or for the lifetime of the subject.

[0240] In some embodiments, a subject who suffers from a thromboembolic disorder or is at risk of suffering from a thromboembolic disorder and is undergoing surgery is administered an intravenous drug delivery formulation on the same day of the surgery. In some embodiments, the intravenous drug delivery formulation is administered between 2 and 10 hours after surgery. In some embodiments, the intravenous drug delivery formulation is administered between 4 and 8 hours after surgery. In some embodiments, the intravenous drug delivery formulation is administered about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, or about 10 hours after surgery.

[0241] In certain embodiments, one or more doses in the initial and subsequent treatment cycles comprise a Factor XI and / or Factor XIa antibody administered subcutaneously at a dose of about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1.0 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 or about 5.0 mg / kg.

[0242] In certain embodiments, one or more doses in the initial and subsequent treatment cycles comprise a factor XI and / or factor XIa antibody (e.g., Antibody 1) administered subcutaneously at a dose of about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 185 mg, about 190 mg, about 195 mg, or about 200 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered subcutaneously at a dose of about 90 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered subcutaneously at a dosage of about 120 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered subcutaneously at a dosage of about 150 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered subcutaneously at a dosage of about 180 mg. In any one of the above embodiments, the factor XI and / or factor XIa antibody is administered subcutaneously monthly.

[0243] In some embodiments, the therapeutically effective dose of factor XI and / or factor XIa antibody (e.g., Antibody 1) after subcutaneous administration ranges from about 75 mg to about 165 mg, about 80 mg to about 160 mg, about 85 mg to about 155 mg, or about 90 mg to about 160 mg. In certain embodiments, the therapeutically effective dose of factor XI and / or factor XIa antibody (e.g., Antibody 1) after subcutaneous administration ranges from about 90 mg to about 160 mg.

[0244] In certain embodiments, one or more doses in the initial and subsequent treatment cycles comprise a Factor XI and / or Factor XIa antibody (e.g., Antibody 1) administered intravenously at a dose of about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1.0 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 or about 5.0 mg / kg.

[0245] In certain embodiments, one or more doses in the initial and subsequent treatment cycles comprise a Factor XI and / or Factor XIa antibody (e.g., Antibody 1) administered intravenously at a dose of about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 185 mg, about 190 mg, about 195 mg, about 200 mg, about 250 mg, about 500 mg, about 750 mg, about 1000 mg, about 1250 mg, about 1500 mg, or about 2000 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered intravenously at a dose of about 30 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered intravenously at a dose of about 60 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered intravenously at a dose of about 75 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered intravenously at a dose of about 150 mg. In some embodiments, the factor XI and / or factor XIa antibody is administered intravenously in a single dose. In some embodiments, FXI / FXIa antibody (eg, antibody 1) is administered in a single dose of about 150 mg. In some embodiments, FXI / FXIa antibody (eg, antibody 1) is administered in a single dose of about 1000 mg. In some embodiments, FXI / FXIa antibody (eg, antibody 1) is administered in a single dose of about 1500 mg. In some embodiments, FXI / FXIa antibody (eg, antibody 1) is administered in a single dose of about 2000 mg.

[0246] In some embodiments, a first dose of a FXI / FXIa antibody (e.g., Antibody 1) is administered in one dose, and a second dose of a FXI / FXIa antibody (e.g., Antibody 1) is administered in a second dose. In certain embodiments, the first dose and the second dose are the same (e.g., 150 mg). In certain embodiments, the first dose is higher than the second dose (e.g., the first dose is about 1000 mg, and the second dose is 150 mg). In certain embodiments, the method further comprises administering a subsequent dose at the same dose as the second dose.

[0247] In certain embodiments, administration of a single dose of FXI / FXIa antibody (e.g., Antibody 1) extends the activated partial thromboplastin time (aPTT) by 400 hours or more, e.g., 400 hours or more, 500 hours or more, or 600 hours or more. In certain embodiments, administration of a single dose of FXI / FXIa antibody (e.g., Antibody 1) extends the activated partial thromboplastin time (aPTT) by about 600 hours.

[0248] Physicians can start the dosage of the disclosed antibody (e.g., Antibody 1) employed in the pharmaceutical composition from a level below the level required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. In general, the effective dosage of the disclosed composition for treating the thromboembolic disorders described herein varies according to many different factors, including the mode of administration, the target site, the physiological state of the patient, other drugs administered, and whether the treatment is preventive or therapeutic. The therapeutic dose can be titrated to optimize safety and efficacy. For systemic administration of the antibody, the dosage is in the range of about 0.01 to 15 mg / kg host body weight. For administration together with the antibody (e.g., subcutaneous or intravenous administration), the dosage may be in the range of 0.1 mg to 5 mg or 1 mg to 600 mg. For example, an anti-FXI / FXIa antibody described herein (e.g., Antibody 1) can be administered at a dose of about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1.0 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 or about 5.0 mg / kg.

[0249] In certain embodiments, the factor XI and / or factor XIa antibody is administered intravenously. For example, in certain embodiments, the factor XI and / or factor XIa antibody is administered by intravenous infusion, for example, using a pre-filled bag, a pre-filled pen or a pre-filled syringe. In certain embodiments, the factor XI and / or factor XIa antibody in the pharmaceutical preparation disclosed herein is diluted before administration. For example, in certain embodiments, the pharmaceutical preparation is diluted with 5% glucose aqueous solution (D5W) and administered intravenously from a bag. Intravenous infusion can last for about one hour (for example, 50 to 80 minutes). In certain embodiments, the bag is connected to a channel including a tube and / or a needle.

[0250] In certain embodiments, the Factor XI and / or Factor XIa antibody is administered parenterally. In certain embodiments, the Factor XI and / or Factor XIa antibody is administered parenterally in one or more doses.

[0251] The types of thromboembolic disorders that may be treated with the Factor XI and / or Factor XIa antibodies or pharmaceutical formulations disclosed herein include, but are not limited to, "thromboembolic" or similar terms as used herein, and may also refer to any number of the following that the anti-FXI and / or FXIa antibodies or antigen-binding fragments thereof disclosed herein may be used to prevent or treat: thromboembolism in subjects with suspected or confirmed arrhythmias (e.g., paroxysmal, persistent or permanent atrial fibrillation or atrial flutter); prevention of stroke in atrial fibrillation (SPAF), a subpopulation of which is AF patients undergoing percutaneous coronary intervention (PCI); treatment of acute venous thromboembolic events (VTE) and extended secondary VTE prevention in patients at high risk for bleeding; transient ischemic stroke; Prevention of cerebral and cardiovascular events in the secondary prevention of a transient ischemic attack (TIA) or non-disabling stroke and of thromboembolic events in heart failure with sinus rhythm; Venous thromboembolism (pediatric VTE) in pediatric subjects; Left atrial clot formation and thromboembolism in subjects undergoing cardioversion for arrhythmias; Thrombosis before, during, and after arrhythmia ablation procedures; Venous thrombosis, which includes but is not limited to the treatment and secondary prevention of deep or superficial vein thrombosis of the lower or upper extremities, abdominal and thoracic vein thrombosis, sinus thrombosis, and jugular vein thrombosis; Thrombosis on any artificial surface of a vein or artery, such as catheters, pacemaker leads, synthetic arterial grafts; Arteriovenous (AV) shunts; Mechanical or biological heart valves or left Ventricular assist devices; pulmonary embolism in patients with or without venous thrombosis; chronic thromboembolic pulmonary hypertension (CTEPH); arterial thrombosis on ruptured atherosclerotic plaques, thrombosis on intra-arterial prostheses or catheters, and thrombosis in seemingly normal arteries, including but not limited to acute coronary syndrome, ST-segment elevation myocardial infarction, non-ST-segment elevation myocardial infarction, unstable angina, stent thrombosis, thrombosis of any artificial surface of the arterial system, and pulmonary artery thrombosis in subjects with or without pulmonary hypertension; thrombosis and thromboembolism in patients undergoing percutaneous coronary intervention (PCI); cardiogenic and cryptogenic stroke; non-CNS systemic embolism (non-CN S systemic embolism); hemorrhagic stroke; thrombosis in patients with invasive and non-invasive cancer malignancies (e.g., gastrointestinal and genitourinary cancers); thrombosis of indwelling catheters; thrombosis and thromboembolism in critically ill patients; cardiac thrombosis and thromboembolism, including but not limited to cardiac thrombosis after myocardial infarction, cardiac thrombosis associated with conditions such as cardiac aneurysms, myocardial fibrosis, cardiac enlargement and insufficiency, myocarditis, and cardiac artificial surfaces; thromboembolism in patients with valvular heart disease with or without atrial fibrillation; thromboembolism of mechanical or biological valve prostheses; thromboembolism in patients with natural or artificial cardiac patches, arterial or venous catheters, after cardiac repair for simple or complex cardiac malformations;Venous thrombosis and thromboembolism following knee replacement surgery, hip replacement surgery, and orthopedic, thoracic, or abdominal surgery; arterial or venous thrombosis following neurosurgery, including intracranial and spinal interventions; congenital or acquired thrombotic tendencies, including but not limited to factor V Leiden, prothrombin mutations, antithrombin III, protein C and protein S deficiency, factor XIII mutations, familial dysfibrinogenemia, congenital plasminogen deficiency, elevated factor XI levels, sickle cell disease, antiphospholipid syndrome, autoimmune diseases, chronic intestinal diseases, nephrotic syndrome, hemolytic uremic disease, myeloproliferative diseases, disseminated intravascular coagulation, paroxysmal nocturnal hemoglobinuria, and heparin-induced thrombocytopenia; thrombosis and thromboembolism in chronic kidney disease; and thrombosis and thromboembolism in patients receiving hemodialysis and patients receiving extracorporeal membrane oxygenation. In certain embodiments, the subject treated with a Factor XI and / or Factor XIa antibody or pharmaceutical formulation disclosed herein is obese (e.g., severely obese, e.g., body mass index (BMI) ≥ 35 kg / m; 2 ). In certain embodiments, the subject treated with the factor XI and / or factor XIa antibody or pharmaceutical formulation disclosed herein is not obese. In certain embodiments, after the same dose of factor XI and / or factor XIa antibody (e.g., antibody 1) is administered to non-obese subjects, obese subjects are associated with lower exposure. In certain embodiments, after the same dose of factor XI and / or factor XIa antibody (e.g., antibody 1) is administered to non-obese subjects, the exposure of obese subjects is reduced by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90%. In certain embodiments, after the same dose of factor XI and / or factor XIa antibody (e.g., antibody 1) is administered to non-obese subjects, obese subjects are associated with a shorter duration of aPTT extension. In certain embodiments, the aPTT prolongation in obese subjects is shortened by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% following administration of the same dose of Factor XI and / or Factor XIa antibody (e.g., Antibody 1) as in non-obese subjects.

[0252] In certain embodiments, factor XI and / or factor XIa antibodies (e.g., antibody 1) are administered to a subject with cancer. It is estimated that venous thromboembolism (VTE) affects 1% to 8% of all cancer patients, and is the second leading cause of death in outpatients receiving chemotherapy. Such VTE in cancer patients is complicated by thrombocytopenia, which is widely present in cancer patients receiving chemotherapy (Bannow et al. (2019). Res. Pract. Throm. Haemost., 2:664-669).

[0253] In certain embodiments, the cancer is a solid tumor. In certain other embodiments, the cancer is brain cancer, bladder cancer, breast cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, leukemia, lung cancer, liver cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, stomach cancer, testicular cancer, or uterine cancer.In other embodiments, the cancer is a vascularized tumor, squamous cell carcinoma, adenocarcinoma, small cell carcinoma, melanoma, glioma, neuroblastoma, sarcoma (e.g., angiosarcoma or chondrosarcoma), laryngeal cancer, parotid cancer, biliary tract cancer, thyroid cancer, acral melanoma, actinic keratosis, acute lymphoblastic leukemia, acute myeloid leukemia, adenoid cystic carcinoma, adenoma, adenosarcoma, adenosquamous carcinoma, anal canal cancer, anal cancer, anorectal cancer, astrocytoma, Bartholin's gland cancer, basal Cell carcinoma, bile duct cancer, bone cancer, bone marrow cancer, bronchial cancer, bronchial adenocarcinoma, carcinoid, bile duct cancer, chondrosarcoma, choroid plexus papilloma / carcinoma, chronic lymphocytic leukemia, chronic myeloid leukemia, clear cell carcinoma, connective tissue cancer, cystadenoma, digestive system cancer, duodenal cancer, endocrine system cancer, endodermal sinus tumor, endometrial hyperplasia, endometrial stromal sarcoma, endometrioid adenocarcinoma, endothelial cell carcinoma, ependymal carcinoma, epithelial cell carcinoma, Ewing's sarcoma sarcoma), cancer of the eye and orbit, cancer of the female genitalia, focal nodular hyperplasia, gallbladder cancer, gastric antrum cancer, gastric fundus cancer, gastrinoma, glioblastoma, glucagonoma, heart cancer, hemangioblastoma, hemangioendothelioma, hemangioma, hepatic adenoma, hepatobiliary cancer, hepatocellular carcinoma, Hodgkin's disease, ileal cancer, insulinoma, intraepithelial neoplasia, intraepithelial squamous cell neoplasm, intrahepatic bile duct cancer, invasive squamous cell carcinoma, jejunal cancer, joint cancer, Kaposi's sarcoma sarcoma), pelvic cancer, large cell carcinoma, colorectal cancer, leiomyosarcoma, malignant lentigo melanoma, lymphoma, male genital cancer, malignant melanoma, malignant mesothelioma, medulloblastoma, medullary epithelioma, meningeal cancer, mesothelioma, metastatic cancer, oral cancer, mucoepidermoid carcinoma, multiple myeloma, muscle cancer, nasal passage cancer, nervous system cancer, neuroepithelial gland carcinoma, nodular melanoma, non-epithelial skin cancer, non-Hodgkin lymphoma, oat cell carcinoma, oligodendroglial carcinoma, oral cancer, osteosarcoma, papillary serous adenocarcinoma, penile cancer, pharyngeal cancer, pituitary tumor, plasma cell tumor, pseudomeatus In some embodiments, the cancer is selected from the group consisting of gastrointestinal cancer and urinary tract cancer.

[0254] In some embodiments, an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) is administered prophylactically, e.g., to prevent coagulation in a subject at risk of thrombosis. In some embodiments, an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) is administered therapeutically, e.g., to treat coagulation in a subject at risk of thrombosis.

[0255] In some embodiments, an anti-FXI / FXIa antibody or antigen-binding fragment thereof (e.g., Antibody 1) is administered to a subject with cancer associated thrombosis (CAT). In certain embodiments, the subject with cancer associated thrombosis has an existing clot. In certain embodiments, the subject has microthrombosis. In certain embodiments, the subject has microthrombosis associated with CAT.

[0256] The CHA2DS2-VASc risk score is a validated and widely used stratification tool that predicts thromboembolic risk in patients with AF and identifies patients who should benefit from anticoagulant therapy (LIP 2011; Camm et al. (2012) EurHeart J 2012; 33: 2719–2747); cumulative evidence suggests that CHA2DS2-VASc is at least as accurate as or possibly better than scores such as CHADS2 in identifying patients who develop stroke and thromboembolism, and is certainly better at identifying 'truly low risk' AF patients. The CHA2DS2-VASc risk score ranges from 0 to a maximum score of 9. In certain embodiments, the CHA2DS2-VASc risk score for subjects treated with a factor XI and / or factor XIa antibody or pharmaceutical formulation disclosed herein is 0-1 for males and 1-2 for females. In certain embodiments, the CHA2DS2-VASc risk score of a subject treated with a factor XI and / or factor XIa antibody or pharmaceutical formulation disclosed herein is ≥ 2 for males and ≥ 3 for females. In certain embodiments, the subject treated with a factor XI and / or factor XIa antibody or pharmaceutical formulation disclosed herein has a CHA2DS2-VASc risk score of ≥ 4 or ≥ 3 and has at least 1 planned concurrent use of an antiplatelet medication (e.g., aspirin and / or a P2Y12 inhibitor), or a CrCl of ≤ 50 ml / min as derived by the Cockcroft-Gault equation.

[0257] Factor XI and / or factor XIa antibodies (e.g., antibody 1) disclosed herein can be used as monotherapy or in combination with one or more therapies. Such combination therapies can be used to treat thromboembolic disorders, such as ischemic stroke (cardiogenic, thrombotic) or systemic embolism, AF, atrial fibrillation stroke prevention (SPAF), deep vein thrombosis, venous thromboembolism, pulmonary embolism, acute coronary syndrome (ACS), acute limb ischemia, chronic thromboembolic pulmonary hypertension or systemic embolism). In certain embodiments, factor XI and / or factor XIa antibodies are used as monotherapy according to the dosing regimen disclosed herein. In other embodiments, factor XI and / or factor XIa antibodies are used in combination with one or more therapies, wherein factor XI and / or factor XIa antibodies are administered according to the dosing regimen disclosed herein, and one or more therapies are administered according to a dosage regimen known to be suitable for treating a specific subject with a specific disorder.

[0258] In some aspects, statin therapy can be used in combination with the FXI / FXIa antibodies and antigen-binding fragments described in the present disclosure or formulations comprising the same (e.g., Antibody 1) to treat patients with thrombotic and / or thromboembolic disorders. In particular aspects, non-limiting examples of therapeutic agents suitable for use in combination with the anti-FXI / FXIa antibodies described herein (e.g., Antibody 1) include thromboprogesterone inhibitors (e.g., aspirin), adenosine diphosphate receptor antagonists (or P2Y12 inhibitors) such as thienopyridines (e.g., clopidogrel and prasugrel) and non-thienopyridines (e.g., ticagrelor and cangrelor), protease activated receptor 1 (PAR1) antagonists (e.g., vorapaxar and atopaxar), and proton pump inhibitors (PPIs) (e.g., omeprazole, diazepam, phenytoin, lansoprazole, dexlansoprazole, rabeprazole, pantoprazole, esomeprazole, and naproxen). The use of a PPI in combination therapy may be appropriate in situations where the subject has or has had a GI disorder, such as prior GI bleeding or a precursor to a peptic ulcer. In one aspect, the subject is being treated with a nonsteroidal anti-inflammatory drug (NSAID) and an anti-FXI / FXIa antibody described herein (e.g., Antibody 1) is administered in combination with a proton pump inhibitor (e.g., omeprazole, diazepam, phenytoin, lansoprazole, dexlansoprazole, rabeprazole, pantoprazole, esomeprazole, and naproxen). In certain embodiments, a direct oral anticoagulant (DOAC) is administered to a subject treated with FXI / FXIa antibodies and antigen-binding fragments, or a formulation comprising the same, such as Antibody 1, after the duration of treatment (e.g., on the same day that treatment ends). In certain embodiments, a vitamin K antagonist (VKA) is administered to a subject treated with FXI / FXIa antibodies and antigen-binding fragments, or a formulation comprising the same, such as Antibody 1, after the duration of treatment (e.g., about 5 days before the end of treatment, or about 3 days before the end of treatment).

[0259] In certain embodiments, the methods of treatment disclosed herein cause a subject or patient to produce a disease response or an improved survival period. For example, in certain embodiments, the disease response is a complete response, a partial response, or a stable condition. In certain embodiments, the improved survival period is an improved progression-free survival period (PFS) or an overall survival period. Improvement (e.g., in PFS) can be determined relative to the period before the treatment of the present disclosure is started. The method for determining the disease response (e.g., complete response, partial response, or stable condition) and patient survival period (e.g., PFS, overall survival period) of BTC (e.g., advanced BTC, metastatic BTC) or biliary tumor therapy is conventional in the art and is considered herein. In some embodiments, after contrast-enhanced computed tomography (CT) or magnetic resonance imaging (MRI) of the affected area (e.g., chest / abdomen and pelvis, covering the area from the upper part of the thoracic inlet to the pubic symphysis) is performed on the treated patient, the disease response is evaluated according to RECIST 1.1.

[0260] Combination therapy and related dosage regimens

[0261] Combination therapy to reverse anticoagulation

[0262] In one aspect, the present invention provides a method for reversing the anticoagulant effect of an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or an antigen-binding fragment thereof in a subject, wherein the isolated antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 150 mg, wherein the method comprises administering to the subject a therapeutically effective amount of recombinant activated factor VII (rFVIIa), thereby reversing the anticoagulant effect. In certain embodiments, such methods are used, for example, in subjects with a high risk of bleeding.

[0263] In some embodiments, temporarily reversing or inhibiting one or more anticoagulant effects of an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) is desirable and / or medically necessary, for example, for subjects with a high risk of bleeding. In some embodiments, bleeding is typically associated with, for example, but not limited to, trauma, surgery, menstruation, or postpartum. Therefore, in these cases, subjects who have been treated with an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) may require rapid and effective therapy to reduce bleeding or reduce the risk of bleeding. In certain embodiments, prolonged bleeding may occur after major trauma or after surgery, such as, but not limited to, surgery involving organs with areas of high fibrinolysis, such as buccal mucosa, nasal mucosa, genital mucosa, or urinary mucosa. Tooth extraction, tonsillectomy, and hysterectomy or prostatectomy are further non-limiting examples of surgeries that result in a high risk of bleeding. In certain embodiments, the simultaneous use of antiplatelet agents, other anticoagulants, and fibrinolytic agents may increase the risk of bleeding. In some embodiments, subjects with a high risk of bleeding can be identified by a history of prior bleeding, such as bleeding during or after surgery, or bleeding while treated with an anticoagulant (e.g., warfarin). In some embodiments, subjects with a high risk of bleeding can be identified by in vitro / ex vivo assays known in the art, such as, but not limited to, assays performed with the subject's blood or plasma: measuring aPTT, assessing biomarkers of the extrinsic coagulation pathway, measuring prothrombin time (PT), thrombin time (TT), rotational thromboelastometry (ROTEM), and / or thromboelastometry. ® )wait.

[0264] In some embodiments, the anticoagulant effect of the anti-FXI / FXIa antibodies or antigen-binding fragments described herein (e.g., Antibody 1) is achieved by administering a therapeutically effective amount of recombinant activated factor VII (rFVIIa), such as a low dose. As used herein, a "low dose" of rFVIIa refers to a therapeutically effective amount of rFVIIa of less than about 15 μg / kg.

[0265] In certain embodiments, a therapeutically effective amount of rFVIIa is from about 0.5 µg / mL to about 14.5 µg / mL, for example, from about 0.5 µg / mL to about 14 µg / mL, from about 0.5 µg / mL to about 13.5 µg / mL, from about 0.5 µg / mL to about 13 µg / mL, from about 0.5 µg / mL to about 12.5 µg / mL, from about 0.5 µg / mL to about 12 µg / mL, from about 0.5 µg / mL to about 11.5 µg / mL, from about 0.5 µg / mL to about 11 µg / mL, from about 0.5 µg / mL to about 10.5 µg / mL, from about 0.5 µg / mL to about 10 µg / mL, from about 0.5 µg / mL to about 9.5 µg / mL, from about 0.5 µg / mL to about 9.5 µg / mL, from about 0.5 µg / mL to about 8.5 µg / mL, from about 0.5 µg / mL to about 8.5 µg / mL. , about 0.5 µg / mL to about 7.5 µg / mL, about 0.5 µg / mL to about 7 µg / mL, about 0.5 µg / mL to about 6.5 µg / mL, about 0.5 µg / mL to about 6 µg / mL, about 0.5 µg / mL to about 5.5 µg / mL, about 0.5 µg / mL to about 5 µg / mL, about 0.5 µg / mL to about 4.5 µg / mL, about 0.5 µg / mL to about 4 µg / mL, about 0.5 µg / mL to about 3.5 µg / mL, about 0.5 µg / mL to about 3 µg / mL, about 0.5 µg / mL to about 2.5 µg / mL, about 0.5 µg / mL to about 2 µg / mL, about 0.5 µg / mL to about 1.5 µg / mL, or 0.5 µg / mL to about 1 µg / mL. In certain embodiments, the therapeutically effective amount of rFVIIa is about 0.5 μg / mL to about 1 μg / mL.

[0266] In certain embodiments, the therapeutically effective amount of rFVIIa is about 14.5 µg / mL, about 14 µg / mL, about 13.5 µg / mL, about 13 µg / mL, about 12.5 µg / mL, about 12 µg / mL, about 11.5 µg / mL, about 11 µg / mL, about 10.5 µg / mL, about 10 µg / mL, about 9.5 µg / mL, about 9 µg / mL, about 8.5 µg / mL, about 8 µg / mL, about 7.5 µg / mL, about 7 µg / mL, about 6.5 µg / mL, about 6 µg / mL, about 5.5 µg / mL, about 5 µg / mL, about 4.5 µg / mL, about 4 µg / mL, about 3.5 µg / mL, about 3 µg / mL, about 2.5 µg / mL, about 2 µg / mL, about 1.5 µg / mL, about 1 In some embodiments, rFVIIa is administered at a dose of about 0.5 μg / mL. In some embodiments, rFVIIa is administered at a dose of about 1 μg / mL.

[0267] In some embodiments, rFVIIa is administered once. In some embodiments, rFVIIa is administered twice. In some embodiments, rFVIIa is administered more than twice, for example, twice, three times, four times, five times or more times.

[0268] In some embodiments, the rFVIIa of therapeutically effective amount is used as needed. As used herein, "using as needed" refers to using rFVIIa until symptoms (such as bleeding) are treated and / or bleeding risk is reduced. In some embodiments, rFVIIa is used for a certain duration, such as the time of surgery. In some embodiments, rFVIIa is used for a certain duration, such as the time of surgery and hospitalization. In some embodiments, rFVIIa is used according to medical needs, such as determined by a physician.

[0269] In some embodiments, rFVIIa is used once per hour. In some embodiments, rFVIIa is used once every two hours. In some embodiments, rFVIIa is used once every four hours. In some embodiments, rFVIIa is used once every six hours. In some embodiments, rFVIIa is used once every eight hours. In some embodiments, rFVIIa is used twice a day, for example, once every twelve hours. In certain embodiments, the frequency of rFVIIa use is reduced along with the reduction of bleeding risk, for example, rFVIIa is used once every two hours in the set period (for example, continues 24 hours or 48 hours), then is used once every four hours in the set period (for example, at most one week).

[0270] In some embodiments, a therapeutically effective amount, such as a low dose, of recombinant activated Factor VII (rFVIIa) is administered to a subject intravenously, such as by intravenous bolus injection.

[0271] In some embodiments, the anti-coagulant effect of the anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) is evaluated using methods known in the art. For example, but not limited to, the anti-coagulant effect can be determined by clotting time (CT), clot formation time (CFT), activated partial thromboplastin time (aPTT), etc. In certain embodiments, administration of a therapeutically effective amount of rFVIIa prolongs the CT of a subject treated with an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1), such that relative to the CT before administration of rFVIIa, the CT is extended by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In certain embodiments, administration of a therapeutically effective amount of rFVIIa prolongs CFT in a subject treated with an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) such that CFT is prolonged by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% relative to CFT prior to administration of rFVIIa.

[0272] In some embodiments, the recombinant activated factor VII (rFVIIa) described herein of therapeutically effective amount can be combined with one or more additional anticoagulant reversal therapies. The non-limiting examples of conventional strategies for reversing anticoagulant effects include (i) using colloid, crystalloid, human plasma or plasma proteins such as albumin to carry out fluid replacement, or (ii) transfusion of concentrated red blood or whole blood. The example of the therapy for reversing anticoagulant effects (for example, in severe emergency situations) includes but is not limited to promoting hemostasis blood components, such as fresh frozen plasma (FFP), prothrombin complex concentrate (PCC) and activated PCC (APCC), such as factor VIII inhibitor bypass activity (FEIBA). In some embodiments, the rFVIIa of therapeutically effective amount and anticoagulant reversal therapy such as emicizumab (ACE910), tranexamic acid, fresh frozen plasma (FFP), Hemoeleven, prothrombin complex concentrate (PCC), activated PCC or FEIBA (a kind of FVIII inhibitor complex) are combined.

[0273] In some embodiments, a therapeutically effective amount of a recombinant activated factor VII (rFVIIa) described herein can be administered in combination with an anti-idiotypic antibody or antigen-binding fragment thereof to an anti-FXI / FXIa antibody or antigen-binding fragment described herein (eg, Antibody 1). Methods for preparing anti-idiotypic antibodies are known in the art.

[0274] Combination antiplatelet therapy

[0275] In one aspect, a combined method for treating a subject is provided herein, comprising administering an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or antigen-binding fragment thereof, such as antibody 1, described herein at a dose of about 150 mg, and administering a therapeutically effective dose of an oral antiplatelet therapy. In some embodiments, the subject suffers from or is at risk of suffering from a thromboembolic disorder. Exemplary antiplatelet therapies include, for example, but not limited to, aspirin, clopidogrel, prasugrel, dipyridamole, ticlopidine, eptifibatide, and ticagrelor. In some embodiments, the antiplatelet therapy is aspirin. In some embodiments, the antiplatelet therapy is clopidogrel. In some embodiments, the antiplatelet therapy is prasugrel. In some embodiments, the antiplatelet therapy is dipyridamole. In some embodiments, the antiplatelet therapy is ticlopidine. In some embodiments, the antiplatelet therapy is eptifibatide. In some embodiments, the antiplatelet therapy is ticagrelor.

[0276] In one aspect, a combined method for treating a subject, such as a subject suffering from a thromboembolic disorder or a subject at risk of suffering from a thromboembolic disorder, is provided herein, comprising administering an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or an antigen-binding fragment thereof, such as antibody 1, described herein at a dose of about 125 nM, about 250 nM, or about 500 nM, and administering about 1 mg / mL aspirin. On the other hand, a combined method for treating a subject, such as a subject suffering from a thromboembolic disorder or a subject at risk of suffering from a thromboembolic disorder, is provided herein, comprising administering an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or an antigen-binding fragment thereof, such as antibody 1, described herein at a dose of about 150 mg, and administering about 100 mg aspirin. In certain embodiments, aspirin is administered orally, such as in the form of a pill or tablet. In certain embodiments, aspirin is low-dose aspirin. In certain embodiments, the dose of aspirin is about 75 mg, about 81 mg, about 100 mg, about 150 mg, about 162 mg, or about 200 mg. In certain embodiments, aspirin is administered daily. In certain embodiments, aspirin is administered twice daily. In certain embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., antibody 1) is administered intravenously. In certain embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., antibody 1) is administered subcutaneously. In certain embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., antibody 1) is administered intravenously in the first dose and subcutaneously in subsequent doses (e.g., the second dose, the third dose, the fourth dose, the fifth dose, the sixth dose, etc.). In certain embodiments, the anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., antibody 1) is administered monthly.

[0277] In one aspect, a combined method for treating a subject, such as a subject suffering from a thromboembolic disorder or a subject at risk of suffering from a thromboembolic disorder, is provided herein, comprising administering an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or an antigen-binding fragment thereof, such as antibody 1, described herein at a dose of about 125 nM, about 250 nM, or about 500 nM, and administering about 3 μM ticagrelor. On the other hand, a combined method for treating a subject, such as a subject suffering from a thromboembolic disorder or a subject at risk of suffering from a thromboembolic disorder, is provided herein, comprising administering an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or an antigen-binding fragment thereof, such as antibody 1, described herein at a dose of about 150 mg, and administering about 60 mg ticagrelor. In certain embodiments, ticagrelor is administered orally, such as in the form of a pill or tablet. In certain embodiments, the dose of ticagrelor is about 60 mg, about 90 mg, or about 180 mg. In certain embodiments, ticagrelor is administered with a loading dose (e.g., about 180 mg), and subsequent administration is administered with a maintenance dose (e.g., about 60 mg or about 90 mg). In certain embodiments, ticagrelor is administered daily. In certain embodiments, ticagrelor is administered twice daily. In certain embodiments, anti-FXI / FXIa antibodies or antigen-binding fragments (e.g., antibody 1) described herein are administered intravenously. In certain embodiments, anti-FXI / FXIa antibodies or antigen-binding fragments (e.g., antibody 1) described herein are administered subcutaneously. In certain embodiments, anti-FXI / FXIa antibodies or antigen-binding fragments (e.g., antibody 1) described herein are administered intravenously in the first dose, and subcutaneously in subsequent doses (e.g., second dose, third dose, fourth dose, fifth dose, sixth dose, etc.). In certain embodiments, anti-FXI / FXIa antibodies or antigen-binding fragments (e.g., antibody 1) described herein are administered monthly.

[0278] In some embodiments, a subject to whom an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., antibody 1) is administered, such as a subject with a thromboembolic disorder or a risk of a thromboembolic disorder, is receiving dual antiplatelet therapy (DAPT). In certain embodiments, the DAPT comprises ticagrelor and aspirin. In some embodiments, about 3 μM ticagrelor, about 1 mg / mL aspirin, and an anti-FXI and / or anti-FXIa antibody described herein, such as antibody 1, are administered to the subject. In certain embodiments, ticagrelor is administered orally, for example, in the form of a pill or tablet. In certain embodiments, ticagrelor is administered daily. In certain embodiments, aspirin is administered orally, for example, in the form of a pill or tablet. In certain embodiments, aspirin is low-dose aspirin. In certain embodiments, aspirin is administered daily. In certain embodiments, an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., antibody 1) is administered intravenously. In certain embodiments, an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) is administered subcutaneously. In certain embodiments, an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) is administered intravenously in a first dose and subcutaneously in subsequent doses (e.g., a second dose, a third dose, a fourth dose, a fifth dose, a sixth dose, etc.). In certain embodiments, an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) is administered monthly.

[0279] In some embodiments, administration of an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1) does not affect platelet aggregation in a subject compared to a subject treated with antiplatelet therapy alone (e.g., aspirin alone or ticagrelor alone). Methods for assessing platelet aggregation are known in the art and include, for example, but not limited to, impedance platelet aggregation assays.

[0280] In some embodiments, for example, when a subject, e.g., a subject administered an anti-FXI / FXIa antibody or antigen-binding fragment described herein (e.g., Antibody 1), has a high risk of bleeding (e.g., as discussed above), the subject can stop receiving antiplatelet therapy and be administered a therapeutically effective amount of recombinant activated Factor VII (rFVIIa), as discussed above.

[0281] Example

[0282] The disclosure now generally described will be more readily understood by reference to the following examples, which are included merely for the purpose of illustrating certain aspects and embodiments of the disclosure and are not intended to limit the scope of the disclosure in any way.

[0283] Example 1 - Treatment of Patients with Vascular Graft Thrombosis

[0284] Purpose and rationale

[0285] The present study was designed to test the effects of increasing concentrations of Antibody 1 compared with vehicle and active controls (abciximab, anti-GP2b3a) on platelet aggregation following stimulation with collagen or thrombin receptor-activating peptide-6 (TRAP-6) and to determine the effects of both agents on thrombin generation.

[0286] method

[0287] Whole blood (6 healthy donors) was obtained in EDTA and citrate tubes. Samples were spiked with vehicle, Antibody 1 (250, 500, or 1000 nM), or Abciximab (50 nM). Platelet aggregation was recorded using a multiplate impedance aggregometer after induction with collagen (1 µg / mL) or TRAP-6 (8 µM). The area under the curve (AUC) for platelet aggregation was determined as arbitrary aggregation*time (AU*min). Thrombin generation was also measured in platelet-rich plasma using a Thrombinoscope CAT (calibrated automated thrombinogram; Stago CH SA) and tissue factor reagent (1 pM) at the above Antibody 1 and Abciximab concentrations; fluorescence was measured by an automated plate reader fluorometer.

[0288] result

[0289] Antibody 1 against collagen ( Figure 1A ) or TRAP-6 ( Figure 1B )-induced platelet aggregation showed no inhibitory or stimulatory effects. In contrast, a significant reduction in collagen-induced platelet aggregation was observed with abciximab. Antibody 1 significantly delayed the lag time of thrombin generation and the time to reach peak concentration; abciximab had no effect on thrombin generation.

[0290] Importantly, these results suggest that targeting FXI / FXIa by antibody 1 does not affect platelet aggregation while inhibiting thrombin generation.

[0291] Example 2 - Antibody 1 does not affect the in vitro effects of two commonly used antiplatelet agents

[0292] Purpose and rationale

[0293] As shown in Example 1 above, Antibody 1 does not affect platelet function. The concomitant use of Antibody 1 with antiplatelet drugs has not been evaluated previously. Therefore, this study was designed to evaluate the effect of increasing concentrations of Antibody 1 on the inhibition of platelet aggregation by aspirin or ticagrelor.

[0294] method

[0295] Two studies were performed in parallel. Blood samples were obtained in EDTA and citrate tubes (six healthy donors in each study). In the first study, whole blood was spiked with vehicle, aspirin (1 mg / mL), or aspirin (1 mg / mL) + Antibody 1 (125, 250, and 500 nM). Platelet aggregation was measured by impedance platelet aggregometry (Multiplate® analyzer, COLtest and ASPItest reagents; Roche Diagnostics International Ltd, Rotkreuz, Switzerland) after induction with collagen (3.2 µg / mL) or arachidonic acid (0.5 mM).

[0296] In the second study, whole blood was spiked with vehicle, ticagrelor (3 µM), or ticagrelor (3 µM) + Antibody 1 (125, 250, and 500 nM). Platelet aggregation was measured using the same method after induction with ADP (6.4 µM) or thrombin receptor activating peptide-6 (TRAP-6, 32 µM).

[0297] result

[0298] In the presence of aspirin, collagen ( Figure 2A ) or arachidonic acid ( Figure 2B )-induced platelet aggregation was inhibited by 60% and 71%, respectively (p<0.05). Antibody 1 at 125, 250, and 500 ng / mL did not attenuate or increase the level of inhibition achieved by aspirin alone.

[0299] In the presence of ticagrelor, ADP ( Figure 3A ) or TRAP-6 ( Figure 3B )-induced platelet aggregation was inhibited by 54% and 31%, respectively (p<0.05). Antibody 1 at 125, 250, and 500 ng / mL did not alter the platelet inhibition achieved with ticagrelor alone.

[0300] in conclusion

[0301] Clinically relevant concentrations of Antibody 1 did not alter the inhibitory effects of aspirin or ticagrelor. These results suggest that pharmacodynamic interactions between Antibody 1 and aspirin or ticagrelor, two commonly used antiplatelet agents, are unlikely.

[0302] Example 3 - Low concentrations of rFVIIa bypass the rotational thromboelastometry (ROTEM) induced by antibody 1 in vitro ® ) Changes in coagulation parameters

[0303] Purpose and rationale

[0304] Low doses of recombinant activated factor VII (rFVIIa) have been used to prevent or stop bleeding in patients with severe factor XI (FXI) deficiency. A similar strategy has been hypothesized as a way to bypass the pharmacodynamic effects of antibody 1. This study was designed to test whether low concentrations of rFVIIa could correct antibody 1-induced changes in coagulation parameters as measured by rotational thromboelastometry (ROTEM) in a whole blood in vitro assay.

[0305] method

[0306] Blood samples from 6 healthy human donors were obtained in citrate tubes. The samples were incubated with 15 and 30 μg / mL of Antibody 1 or vehicle at 37°C for 10 minutes. The samples were then spiked with 0.5 and 1 μg / mL of rFVIIa or vehicle control. ® The delta analyzer monitored clot formation, and non-activated thromboelastometry (NATEM) testing was performed using a recalcification reagent. CT (clotting time in seconds), CFT (clot formation time in seconds), alpha angle, etc. were measured and compared with reference ranges provided by the manufacturer (Pentapharm GmbH, Munich, Germany).

[0307] result

[0308] Compared to baseline, concentrations of 15 and 30 µg / mL of Antibody 1 prolonged CT by 61% and 64%, prolonged CFT by 37% and 32%, and decreased α angle by 10% and 14%, respectively. Addition of 0.5 and 1.0 µg / mL of rFVIIa to samples spiked with Antibody 1 shortened CT by 21% and 38%, shortened CFT by 33% and 49%, and increased α angle by 29% and 47%, respectively. CT parameters are presented in Figure 4A , and the CFT parameters are presented in Figure 4B When rFVIIa was added, NATEM parameters remained within the normal reference range.

[0309] in conclusion

[0310] Low concentrations of rFVIIa (0.5 to 1 μg / mL) were able to correct the effects of Antibody 1, as assessed by rotational thromboelastometry. Furthermore, normalization without overcorrection of ROTEM coagulation parameters suggests that the tested rFVIIa concentrations are unlikely to result in a hypercoagulable state. These data support the use of low doses of rFVIIa for bleeding management in patients treated with Antibody 1.

Claims

1. A method for reversing the anticoagulant effect of an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or an antigen-binding fragment thereof in a subject, wherein the isolated antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 150 mg, wherein the method comprises administering to the subject a therapeutically effective amount of recombinant activated factor VII (rFVIIa), thereby reversing the anticoagulant effect.

2. The method of claim 1, wherein the therapeutically effective amount of rFVIIa is a low dose.

3. The method of claim 1 or 2, wherein the therapeutically effective amount of rFVIIa is between 0.5 and 1 μg / mL.

4. The method of any one of claims 1-3, wherein the therapeutically effective amount of rFVIIa is administered once.

5. The method of any one of claims 1-3, wherein the therapeutically effective amount of rFVIIa is administered on demand.

6. The method of any one of claims 1-5, wherein administration of the therapeutically effective amount of rFVIIa prolongs the clotting time (CT) relative to the CT before administration of the therapeutically effective amount.

7. The method of any one of claims 1-5, wherein administration of the therapeutically effective amount of rFVIIa prolongs clot formation time (CFT) relative to CFT prior to administration of the therapeutically effective amount.

8. The method of claim 6 or 7, wherein the CT or CFT is determined by rotational thromboelastometry (ROTEM) in a whole blood assay.

9. The method of claim 8, wherein the whole blood assay is an ex vivo or in vitro assay.

10. The method of any one of claims 1-9, wherein the method further comprises applying to the subject one of: (i) fluid replacement using colloids, crystalloids, human plasma, or plasma proteins such as albumin; (ii) transfusion of concentrated red blood or whole blood; (iii) administration of fresh frozen plasma (FFP), prothrombin complex concentrate (PCC), activated PCC (APCC), such as a Factor VIII inhibitor; or (iv) an anti-idiotypic antibody to the isolated anti-FXI and / or anti-FXIa antibody or an antigen-binding fragment thereof.

11. A method of treating a disease or condition in a subject in need thereof, the method comprising administering to the subject a dose of about 150 mg of an isolated anti-factor XI (FXI) and / or anti-activated factor XI (FXIa) antibody or antigen-binding fragment thereof, and administering (a) about 1 mg / mL aspirin or (b) about 3 µM ticagrelor.

12. The method of claim 11, wherein the aspirin or ticagrelor is administered prior to administration of the antibody or antigen-binding fragment thereof.

13. The method of claim 11, wherein the aspirin or ticagrelor is administered concurrently with the administration of the antibody or antigen-binding fragment thereof.

14. The method of claim 11, wherein the aspirin or ticagrelor is administered after administration of the antibody or antigen-binding fragment thereof.

15. The method of any one of claims 11-14, wherein administration of the antibody or antigen-binding fragment thereof does not affect platelet aggregation in the subject compared to the subject treated with aspirin or ticagrelor alone.

16. The method of claim 15, wherein the platelet aggregation is measured by impedance platelet aggregation assay.

17. The method of claim 16, wherein the platelet aggregation is induced by collagen, adenosine 5'-diphosphate (ADP) or thrombin receptor activating peptide-6 (TRAP-6).

18. The method of any one of claims 15-17, wherein the platelet aggregation is determined ex vivo or in vitro.

19. The method of any one of claims 1-18, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) in SEQ ID NO: 9 or 29, wherein the VH comprises complementarity determining regions HCDR1, HCDR2, and HCDR3; and a light chain variable region (VL) in SEQ ID NO: 19 or 39, wherein the VL comprises complementarity determining regions LCDR1, LCDR2, LCDR3.

20. The method of any one of claims 1-19, wherein the antibody or antigen-binding fragment thereof comprises: i. heavy chain variable region CDR1 of SEQ ID NO: 23; heavy chain variable region CDR2 of SEQ ID NO: 24; heavy chain variable region CDR3 of SEQ ID NO: 25; light chain variable region CDR1 of SEQ ID NO: 33; light chain variable region CDR2 of SEQ ID NO: 34; and light chain variable region CDR3 of SEQ ID NO: 35; ii. heavy chain variable region CDR1 of SEQ ID NO: 26; heavy chain variable region CDR2 of SEQ ID NO: 27; heavy chain variable region CDR3 of SEQ ID NO: 28; light chain variable region CDR1 of SEQ ID NO: 36; light chain variable region CDR2 of SEQ ID NO: 37 (or the amino acid sequence of KNY); and light chain variable region CDR3 of SEQ ID NO: 38; iii. the heavy chain variable region CDR1 of SEQ ID NO: 43; the heavy chain variable region CDR2 of SEQ ID NO: 44; the heavy chain variable region CDR3 of SEQ ID NO: 45; the light chain variable region CDR1 of SEQ ID NO: 47; the light chain variable region CDR2 of SEQ ID NO: 37 (or the amino acid sequence of KNY); and the light chain variable region CDR3 of SEQ ID NO: 15; or iv. the heavy chain variable region CDR1 of SEQ ID NO: 46; the heavy chain variable region CDR2 of SEQ ID NO: 4; the heavy chain variable region CDR3 of SEQ ID NO: 5; the light chain variable region CDR1 of SEQ ID NO: 33; the light chain variable region CDR2 of SEQ ID NO: 14; and the light chain variable region CDR3 of SEQ ID NO:

15.

21. The method of any one of claims 1-20, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) selected from the group consisting of SEQ ID NOs: 9, 29, and a VH with 90% identity thereto; and a light chain variable region (VL) selected from the group consisting of SEQ ID NOs: 19, 39, and a VL with 90% identity thereto.

22. The method of any one of claims 1-21, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) selected from the group consisting of SEQ ID NOs: 9 and 29; and a light chain variable region (VL) selected from the group consisting of SEQ ID NOs: 19 and 39.

23. The method of any one of claims 1-22, wherein the antibody comprises a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 31, 11, and a heavy chain with 90% identity thereto; and a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 41, 21, and a light chain with 90% identity thereto.

24. The method of any one of claims 1-23, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31, and a light chain comprising the amino acid sequence of SEQ ID NO:

41.

25. The method of any one of claims 1-24, wherein the antibody is a human monoclonal antibody.

26. The method of claim 25, wherein the antibody is of the human IgG1 isotype.

27. The method of claim 25 or 26, wherein the antibody comprises D265A and P329A substitutions in the Fc domain.

28. The method of any one of claims 1-27, wherein the antibody or antigen-binding fragment thereof is administered intravenously.

29. The method of any one of claims 1-28, wherein the antibody or antigen-binding fragment thereof is administered subcutaneously.

30. The method of any one of claims 1-29, wherein the antibody or antigen-binding fragment thereof is administered in a drug delivery formulation comprising a histidine buffer at a concentration of about 20 mM.

31. The method of any one of claims 1-30, wherein the antibody or antigen-binding fragment thereof is administered in a drug delivery formulation comprising sucrose at a concentration of about 220 mM.

32. The method of any one of claims 1-31, wherein the antibody or antigen-binding fragment thereof is administered in a drug delivery formulation comprising polysorbate 20 at a concentration of about 0.04%.

33. The method of any one of claims 1-32, wherein the antibody or antigen-binding fragment thereof is administered in a drug delivery formulation at pH 5.

5.

34. The method of any one of claims 1-33, wherein when the antibody or antigen-binding fragment thereof is administered in an intravenous drug delivery formulation, the intravenous drug delivery formulation further comprises about 5% dextrose.

35. The method of any one of claims 1-34, wherein the subject has or is at risk of having a thromboembolic disorder.

36. The method of claim 35, wherein the thromboembolic disorder is selected from the group consisting of atrial fibrillation or atrial flutter, transient ischemic attack, ischemic stroke, thromboembolic stroke, hemorrhagic stroke, venous thromboembolism (VTE), pediatric VTE, systemic embolism, non-central nervous system embolism, myocardial infarction, deep vein thrombosis, severe protein S deficiency, cerebrovascular accident, and cancer.

37. The method of any one of claims 1-36, wherein a first dose of the antibody or antigen-binding fragment thereof is administered intravenously and a second dose of the antibody or antigen-binding fragment thereof is administered subcutaneously.

38. The method of claim 37, further comprising subcutaneously administering one or more additional doses of the antibody or antigen-binding fragment thereof after administering the second dose.

39. The method of any one of claims 1-38, wherein the antibody or antigen-binding fragment thereof is administered about once a month.

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