Factor XI antibody and method of use

Abstract

This treatment offers comparable efficacy to existing therapies but with reduced bleeding risk, and provides a safer treatment that can reduce thromboembolic complications of AF, such as stroke, systemic embolism, cognitive decline, and mortality. [Solution] The present invention provides a monoclonal antibody that binds to human coagulation factor XI and XIa (activated factor XI) and its antigen-binding fragment, as well as a pharmaceutical composition containing the same, and a treatment method comprising the step of administering the same.

Description

Technical Field

[0001] This application claims the benefit of each of U.S. Provisional Application No. 62 / 184,955, filed Jun. 26, 2015, and U.S. Provisional Application No. 62 / 341,568, filed May 25, 2016, each of which is hereby incorporated by reference in its entirety.

[0002] This application was electronically filed in ASCII format and contains a Sequence Listing that is hereby incorporated by reference in its entirety. The ASCII copy, created on Jun. 23, 2016, is named "PAT056955-WO-PCT_SL.txt" and is 45,685 bytes in size.

[0003] Background Thrombosis refers to the formation of a thrombus inside a blood vessel following a combination of hereditary risk factors known as thrombotic predisposition or hypercoagulable state and acquired risk factors. Damage to the blood vessel wall, congestion, increased platelet reactivity, and activation of coagulation factors are some of the root features of thrombosis. Thrombosis can occur in either the venous circulation or the arterial circulation, resulting in the onset of deep vein thrombosis (DVT), pulmonary embolism, and stroke. When a thrombus occurs in the arterial system, ischemia occurs downstream, resulting in acute coronary syndrome (ACS), ischemic stroke, and acute lower limb ischemia. Thrombosis in the venous system typically results in deep vein thrombosis, pulmonary embolism, and chronic thromboembolic pulmonary hypertension. Coagulation can also form inside the left atrial appendage of patients with atrial fibrillation (AF) and break off These blood clots can lead to serious complications, namely thromboembolic stroke and systemic stroke. This can result in embolism. Low molecular weight heparin (LMWH), thrombin inhibitor —, and all currently available antithrombotic drugs, including factor Xa (FXa) inhibitors, Associated with a significant risk of bleeding (Weitz JI (2010) Thromb. Haemost. 103, 62). Hemostasis The development of antithrombotic agents that do not affect and therefore do not result in bleeding complications is It is strongly desired.

[0004] Current anticoagulants are administered by injection or orally. Injectable anticoagulant LMWH It is widely used and has a superior therapeutic profile compared to unfractionated heparin, which was previously used. This has brought about improvements. Over the past several decades, the most commonly used oral anticoagulants have been: It was warfarin. Warfarin has a narrow therapeutic window, and therefore, the clotting state... It requires frequent monitoring and exhibits various drug interactions. More recently, Orally available direct FXa and thrombin inhibitors enter the anticoagulant market. Its application is increasing.

[0005] LMWH, FXa inhibitor, and thrombin inhibitor are all used in postoperative veins. Prevention of thromboembolic diseases, management of spontaneous DVT and pulmonary embolism, and in atrial fibrillation These anticoagulants are effective in preventing strokes. However, these anticoagulants are also a step down from older drugs. Bleeding complications generally equivalent to those observed with rufarin and unfractionated heparin. It is also associated with the disease. In the ADVANCE-2 clinical trial, the FXa inhibitor apixaban ( Eliquis) is used in patients after total knee arthroplasty, and enoxapari is a LMWH (Leading Membrane Heat Stretch). It was compared to . Acute apixaban therapy is used to prevent venous thromboembolic disease, and enoki Although more effective than saparin, all drugs were associated with a significant risk of bleeding. Targeted bleeding occurred in 4% of patients receiving apixaban and enoxaparin. It occurred in 5% of patients treated with [the drug] (Lassen, MR, et al. (2009) N. Engl.). J. Med. 361, 594).

[0006] In the RE-LY trial, the direct thrombin inhibitor dabigatran (Prada) xa) In patients with atrial fibrillation and a risk of stroke, compared to warfarin (Connolly, SJ, et al. (2009) N. Engl. J. Med. 361, 1139). Chronic dabigatran The therapy was associated with a significant reduction in the risk of stroke or systemic embolism. However, massive bleeding was associated with... Complications occurred in 3.1% of patients receiving 150 mg of dabigatran daily, and This occurred in 3.4% of patients receiving rufarin (p=0.31).

[0007] Atrial fibrillation (AF) remains the most common cardiac arrhythmia in clinical practice. It accounts for about one-third of hospitalizations due to tuning problems. Currently, AF accounts for 60 in Europe. It is estimated to affect over 0,000 patients, and approximately 2.3 million in the United States. Due to the increasing proportion of the aging population, this number continues to grow rapidly. (People over 65 years old) It is estimated that approximately 5% of the population and 10% of people over 80 years old will develop atrial fibrillation (AF). However, the prevalence of AF is increasing beyond what can be explained by age alone. Hypertension, congestive heart failure, left ventricular hypertrophy, coronary artery disease, and diabetes, as well as obstructive sleep disorders. Risk factors for AF, such as sleep apnea, are also increasing. In fact, among Westerners, this is expected to increase two to three times over the next 30 years (Kannel). and Benjamin (2008) Med Clin North Am. 2008; 92:17-40; Bunch, et al. (2012) JI nnovations of Card Rhythm Manag 2012; 3: 855-63).

[0008] The primary risk factor for AF is a 4-5 times increased risk of embolic stroke. This is attributed to its association with AF. The risk of stroke increases sharply with age, rising to 23.5% for those aged 80-89. It is significant. AF is associated with a doubling of mortality in both sexes (Kannel and Benjamin). (min 2008). AF is also independently associated with cognitive decline and all forms of dementia. (Marzona, et al. (2012) CMAJ 2012; 184: 329-36; Geita et al 2013; Bunch et al. (al 2012).

[0009] Most patients with AF need to take precautions throughout their lives to prevent cardioembolic stroke and systemic embolism. The patient is required to undergo continuous anticoagulation therapy. The CHA2DS2-VASc risk score has been validated. Therefore, to predict the risk of thromboembolism in patients with atrial fibrillation and to determine which patients will benefit from anticoagulation therapy. It is a widely used stratification tool for identifying individuals (LIP 2011; Camm, et al. (20 12) Eur Heart J 2012; 33: 2719-2747), the accumulation of evidence shows that CHA2DS2-VASc To identify patients who develop stroke and thromboembolism, scores such as CHADS2 and At least as accurate, or perhaps even better, with autofocus. It is definitively superior in identifying patients who are "truly at low risk". This indicates that 85–90% of AF patients will require anticoagulant therapy.

[0010] The efficacy of vitamin K antagonists (VKAs) in reducing stroke and systemic embolism. A meta-analysis, including six studies, that assessed the effects showed that the risk of stroke incidence was... A highly significant reduction was observed (the relative reduction in the risk of stroke was 67%). The mortality rate was significantly reduced (26%) with dose-adjusted VKA compared to the control group. (Hart, Pearce, and Aguilar (2007) Ann Intern Med 2007; 146:857-867). 2-3 The target INR (international normalized ratio) value is related to the best profit-risk ratio. (Hylek et al (2003) N Engl J Med; 349:1019-1026), international guidelines and domestic guidelines It is widely adopted in the IDE.

[0011] In recent years, new oral anticoagulants (NOA), also known as direct oral anticoagulants (DOACs), have emerged. C) has been approved and introduced into clinical practice. These drugs reduce the risk of thromboembolic disease. In reducing the dosage, it is at least as effective as warfarin, or even better. (Connolly, et al. (2009) N Engl J Med; 361:1139-51; Connolly, et al. (2011) N Engl J Med; 364:806-17; Patel, et al. (2011) N Engl J Med 2011; 365:883-91). NOACs also have a lower risk of the most serious complications of warfarin, namely hemorrhagic stroke and cerebral infarction. It was also associated with a significant reduction in internal bleeding. Massive bleeding events were associated with well-administered warfarin therapy and They were equivalent or slightly less. In addition, NOACs were smaller than warfarin. In relation to the possibility of drug-drug interactions, use without routine monitoring is... This is expected to facilitate their use in routine medical practice.

[0012] Despite recent improvements, the risk of bleeding associated with anticoagulant use remains high. It goes as far as [number missing]. For example, the annual incidence rates of massive bleeding and clinically relevant non-massive bleeding are 14. The figure was 9%, and in the ROCKET study, among patients treated with rivaroxaban, The annual incidence of major bleeding events was 3.6% (Patel et al 2011). In patients whose HAS Bled risk score is defined as ≥3, massive bleeding The annual incidence rate was >5% (Gallego, et al. (2012) Carc Arrhythm Electrophysiol). (5:312-318). Massive bleeding is a particularly significant clinical outcome, for example, ROCKE In the T study, when massive bleeding occurred, the all-cause mortality rate was 20% in the rivaroxaban group. The rates were 0.4% in the 0.4% group and 26.1% in the warfarin group. In the event of a massive bleeding event, 4.7% and 5% of patients in the rivaroxaban group and the warfarin group, respectively. Stroke and systemic embolism occurred in 4% of cases (Piccini, et al. (2014) Eur Heart J; 35 (1873-80). Hospitalization, intravenous administration of blood products, and the use of blood supply sources may also contribute to the occurrence of massive bleeding. The patient was severely affected. The risk of bleeding is also higher in eligible patients who do not receive anticoagulants. This is also a major reason. 182 hospitals in 35 countries and 5,333 outpatients with AF and Euro Heart Survey on Art, including data from hospitalized AF patients. In oral fibrillation, 67% of eligible patients received oral anticoagulant at discharge. They were only given a coagulant (Nieuwlaat, et al (2005) Eur Heart J;26, 2422-243) 4).

[0013] Therefore, it has comparable efficacy to existing treatments but reduces the risk of bleeding, stroke, all It may reduce thromboembolic complications of AF, such as physical embolism, cognitive decline, and mortality. There is a great unmet medical need for safer treatments.

[0014] overview The present invention relates to human coagulation factor XI and XIa (activated factor XI) (hereinafter, in the case of Therefore, the monochromatic (referred to as "FXI", "FXIa", and similar terms) A nal antibody, a pharmaceutical composition containing the same, and a treatment method comprising the step of administering the same. Regarding thrombosis or thromboembolic diseases / disorders (e.g., thrombotic stroke, atrial fibrillation, atrial Prevention of stroke in fibrillation (SPAF), deep vein thrombosis, venous thromboembolism, pulmonary embolism, Acute coronary syndrome (ACS), ischemic stroke, acute lower limb ischemia, chronic thromboembolic pulmonary hypertension It is effective in preventing and treating (systemic embolism), but carries a risk of bleeding. If we develop antithrombotic agents that carry only a minimal risk of bleeding, then the requirements will be met. This will likely fulfill major, unmet medical needs.

[0015] In specific embodiments, the antibodies presented herein (e.g., human monoclonal antibodies, etc.) Melanomoclonal antibodies (humanized monoclonal antibodies) are human FXIa and FXI It binds to the catalytic domain (CD) with similar high affinity, and within FXIa, the inactive It induces protease domain conformation.

[0016] The isolated anti-FXI antibodies and / or anti-FXIa antibodies described herein, for example, Full-length IgG with the two binding sites described in the specification is FXI and / or FXI a is an equilibrium dissociation constant (K) less than or equal to 100 pM. D ) are joined together. For example, as described herein The isolated antibodies were found to be human FXI and / or FXIa at concentrations of ≤100 pM and ≤50 pM. K ≤ 45 pM, ≤ 40 pM, ≤ 35 pM, ≤ 20 pM, or ≤ 10 pM D It can bind to human FXI. More specifically, the isolated antibodies described herein can also bind to human FXI. and / or FXIa, surface plasmon resonance (SPR), e.g., BIACORE( (Trademark) assays measuring 34 pM or less, or equilibrium titration assays in solution (SE K ≤ 4 pM as measured by T) D It is also possible to combine them; and also, crab quizzes FXI and / or FXIa are measured by the BIACORE® assay. K is below 53 pM, or below 4 pM as measured by SET. D It is also possible to combine them using this method. It is possible. In specific embodiments, the isolated antibody described herein (e.g., NOV1401) ) can be detected in human FXI and FXIa, for example, by a solution equilibrium titration assay (SET). The measured concentrations are approximately 5 pM (e.g., 4.7 pM) and 2 pM (e.g., 1.7 pM), respectively. Apparent K below 3 pM D They are joined together. In specific embodiments, as described herein. Anti-FXI / FXIa antibodies are used against FXI / FXIa in cynomolgus monkeys, for example, in SET (e.g., in the SET). Approximately 12.5 (±6.6) pM relative to FXIa, as measured by (see Example 2). and an apparent K of approximately 5.0 (±0.7) pM D They are joined together. In a specific embodiment, this The anti-FXI / FXIa antibodies described in the specification are rabbit FXI and / or FXIa In addition, there is a K of approximately 20 (±2) nM. D They are joined together. In specific embodiments, the anti described herein FXI / FXIa antibodies are used in humans, cynomolgus monkeys, and rabbits for FXI and / or F It binds to XIa, but does not specifically bind to mouse or rat FXI.

[0017] Isolated anti-FXI antigen-binding fragments and / or anti-FXIa antigen-binding fragments described herein. Compatibility fragments, for example, Fab fragments and other fragments containing one binding site, are FXI and / or FXIa, equilibrium dissociation constant (K) less than or equal to 10 nM D ) is used to join them. For example, The isolated antigen-binding fragments described herein are used in human FXI and / or FXIa. , 10nM or less, 5nM or less, 1nM or less, 500pM or less, 305pM or less, 62pM The following KDs can be bound. More specifically, isolated antigen-binding fragments as described herein. It also binds to human FXI and / or FXIa with a KD of 305 pM or less. It is also possible.

[0018] This invention relates to an isolated antibody or so which binds to FXIa in humans, rabbits, and cynomolgus monkeys. The present invention relates to antigen-binding fragments of FXI and / or FXIa catalyst domaine. Specifically, within the active site region, an isolated antibody or its antigen-binding fragment that binds to the surface of the active site region It also relates to this.

[0019] The present invention also conjugates to FXI and / or FXIa and the antibodies listed in Table 1 ( For example, isolated antibodies or their antigen binding that further compete for binding to NOV1401) This also relates to sex fragments. Between antibodies and / or their antigen-binding fragments as described herein. "Competition" in this context means that both antibodies (or their binding fragments) are the same or overlapping FXI antibodies. To bind to a pitope and / or FXIa epitope (for example, as well as known to those skilled in the art) This means (determined in a competitive binding assay by one of the methods). The antibody or its antigen-binding fragment used in the details is the competing antibody or its antigen-binding The fragment has the same FXI epitope and / or F as the antibody or antigen-binding fragment of the present invention. XIa epitope, or overlapping FXI epitope and / or FXIa epitope The FXI antibody and / or FXIa antibody of the present invention, as long as they do not bind to a pitope. Or compete with antigen-binding fragments (e.g., NOV1401 or NOV1090) i. The competitive antibody or its antigen-binding fragment used herein is (i) the antibody of the present invention Alternatively, the antigen-binding fragment is sterically blocked from binding to its target (for example, the competition The antibody binds to nearby, non-overlapping FXI epitopes and / or FXIa epitopes. Furthermore, the antibody or antigen-binding fragment of the present invention is physically prevented from binding to its target. (ii) different, non-overlapping FXI epitopes and / or F It binds to the XIa epitope and to the FXI protein and / or FXIa protein. The FXI antibody and / or FXIa antibody or antigen-binding fragment of the invention conforms In the absence of conformational changes, the conformational changes are no longer able to bond in the same way. It does not contain a competitive antibody or its antigen-binding fragment that induces a change in the protein.

[0020] In one embodiment, the antibodies listed in Table 1 are conjugated to FXI and / or FXIa. The isolated antibodies or their antigen-binding fragments that further compete for binding with the aforementioned antibodies are listed in Table 1. It binds to most of the amino acids of the epitope to which it binds. In another embodiment, FXI and / or binds to FXIa and further competes for binding with the antibodies listed in Table 1. The isolated antibody or its antigen-binding fragment binds to all of the epitopes to which the antibody binds, as shown in Table 1. To combine.

[0021] In one embodiment, the isolated antibody or its antigen-binding fragment is active to FXI(FXIa). It binds, but when it binds to the active FXI (FXIa) catalytic domain, it binds to FXIa. The information is changed to an inactive conformation. In another embodiment, the previous The isolated antibody or its antigen-binding fragment is the four N-terminus of the inactive conformation. The residues, loops 145, 188, and 220, compared to the active conformation, This induces further changes that shift and / or become irregular.

[0022] In one embodiment, the isolated antibody or its antigen-binding fragment is FXI (e.g., human FXI). ) binds, but when it binds to FXI, the FXI catalytic domain becomes loops 145, 188, and 220 is an active conformation that is as regular as the structure within the FXIa catalytic domain. To prevent the removal of the omen.

[0023] In one embodiment, the isolated antibody or its antigen-binding fragment binds to FXI, but FXI When it binds, the FXI catalytic domain consists of four residues at the N-terminus, loops 145 and 188, and The active conformation in which 220 is regular as in the structure of the FXIa catalytic domain To prevent the removal of the n.

[0024] In one embodiment, the isolated antibody or its antigen-binding fragment binds to FXI, but FXI When it binds, it induces conformational changes within the zymogen structure, and binds to FXIa. Inhibitory FXI conformation observed when and closely related to inhibitory FXI conformation By further introducing the conformation, the FXI catalytic domain becomes active conformation To prevent the release of urine.

[0025] In one embodiment, the isolated antibody or its antigen-binding fragment is FXI and / or FXI It binds to a, but binds to FXI and / or FXIa, and FXI and / or FXI When the catalytic domain of a forms an antibody:antigen complex, loops 145, 188, and 2 20. The uncomplexed and relative structures of the catalytic domain of the active factor XI (FXIa). This causes shift and / or loss of orientation when compared.

[0026] In one embodiment, the isolated antibody or its antigen-binding fragment is FXI and / or FXI It binds to a, but binds to FXI and / or FXIa, and FXI and / or FXI When the catalytic domain of a forms an antibody:antigen complex, the four residues at the N-terminus, loop 145 Complexation of the catalytic domains of the active factor XI (FXIa) at 188 and 220. This causes a shift and / or loss of orientation compared to a structure without this modification.

[0027] In one embodiment, the isolated antibody or its antigen-binding fragment is active to FXI(FXIa). It binds to the FXI(FXIa) catalytic domain, and its conformation is in loop 145. , 188, and 220 are shifted compared to the active conformation, and / or are distributed It transforms into an inactive conformation in which directional properties are lost.

[0028] In one embodiment, the isolated antibody or its antigen-binding fragment binds to FXI, but thymo It induces conformational changes within the gene structure, thereby when binding to FXIa. Observed inhibitory FXI conformation and inhibitory FXI conformation closely related By causing this, the catalytic domain is prevented from adopting the active conformation. Stop.

[0029] The present invention also provides the same epitone as the antibodies listed in Table 1 (e.g., NOV1401). The invention also relates to isolated antibodies that bind to the antigen or their antigen-binding fragments.

[0030] The binding affinity of the isolated antibodies and antigen-binding fragments described herein can be determined by solution equilibrium titration (SET). The SET method is known in the art and is described in further detail below. Alternatively, the binding affinity of the isolated antibodies or fragments described herein can be determined, for example, by surface plasmon resonance measurements in a BIACORE™ assay. Methods for BIACORE™ reaction rate assays are known in the art and are described in further detail below. The binding affinity of the isolated antibodies and antigen-binding fragments described herein can be determined by solution equilibrium titration (SET). The SET method is known in the art and is described in further detail below. Alternatively, the binding affinity of the isolated antibodies or fragments described herein can be determined, for example, by surface plasmon resonance measurements in a BIACORE™ assay. Methods for BIACORE™ reaction rate assays are known in the art and are described in further detail below. The binding affinity of the isolated antibodies and antigen-binding fragments described herein can be determined by solution equilibrium titration (SET). The SET method is known in the art and is described in further detail below. Alternatively, the binding affinity of the isolated antibodies or fragments described herein can be determined, for example, by surface plasmon resonance measurements in a BIACORE™ assay. Methods for BIACORE™ reaction rate assays are known in the art and are described in further detail below. The binding affinity of the isolated antibodies and antigen-binding fragments described herein can be determined by solution equilibrium titration (SET). The SET method is known in the art and is described in further detail below. Alternatively, the binding affinity of the isolated antibodies or fragments described herein can be determined, for example, by surface plasmon resonance measurements in a BIACORE™ assay. Methods for BIACORE™ reaction rate assays are known in the art and are described in further detail below. The binding affinity of the isolated antibodies and antigen-binding fragments described herein can be determined by solution equilibrium titration (SET). The SET method is known in the art and is described in further detail below. Alternatively, the binding affinity of the isolated antibodies or fragments described herein can be determined, for example, by surface plasmon resonance measurements in a BIACORE™ assay. Methods for BIACORE™ reaction rate assays are known in the art and are described in further detail below. The binding affinity of the isolated antibodies and antigen-binding fragments described herein can be determined by solution equilibrium titration (SET). The SET method is known in the art and is described in further detail below. Alternatively, the binding affinity of the isolated antibodies or fragments described herein can be determined, for example, by surface plasmon resonance measurements in a BIACORE™ assay. Methods for BIACORE™ reaction rate assays are known in the art and are described in further detail below.

[0031] The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway.

[0032] The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. The isolated anti-FXI antibodies and / or isolated FXIa antibodies and antigen-binding fragments described herein can be used to inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin, and / or the binding to platelet receptors, thereby preventing the activation of the intrinsic coagulation pathway and / or the common coagulation pathway. IC 50 The isolated antibodies or antigen-binding fragments described herein can inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin at an IC The isolated antibodies or antigen-binding fragments described herein can inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin at an IC The isolated antibodies or antigen-binding fragments described herein can inhibit the direct or indirect activation of factor IX (also known as FIX), factor X (FX), and / or thrombin at an IC 50nM or less, 35nM or less, 25nM or less, 10nM or less, or 5.2nM or less I C 50 It can be inhibited by the isolated antibody or its antigen described herein. More specifically, the isolated antibody or antigen described herein. The binding fragments are factor IX (also known as FIX), factor X (FX), and Direct or indirect activation of thrombin, less than 100 nM, 50 nM The following ICs have a minimum impedance of 35nM, 25nM, 20nM, or 18nM. 50 hindered It may be harmful. More specifically, the isolated antibodies or their antigen-binding fragments described herein may be harmful. , factor IX (also known as FIX), factor X (FX), and / or Direct or indirect activation of thrombin, below 100 nM, below 50 nM, below 35 nM ICs with a minimum impedance of M, 25nM or less, 10nM or less, or 5nM or less. 50 This can be inhibited. Specifically In a typical embodiment, the anti-FXI / FXIa antibody or its antigen-binding properties as described herein The fragments mediated the activation of its natural substrate, FIX, via FXIa, at concentrations of less than 2 nM. For example, a 1.8nM IC 50 It inhibits it.

[0033] Isolated anti-FXI antibodies and / or isolated anti-FXIa antibodies, or their antigen-binding decryption. Using this method, for example, inhibiting the activation of FIX mediated by FXI and / or FXIa. Through this, the intrinsic coagulation pathway and / or common coagulation pathway are inhibited (e.g., these activations It can block sexualization. Therefore, isolated anti-FXI / FXIa antibodies, or Antigen-binding fragments can be used to prevent blood clots or the expansion of blood clots. Isolated anti The body or its antigen-binding fragment inhibits the activation of FIX mediated by FXI. , prevention and treatment of coagulation disorders such as deep vein thrombosis and stroke (e.g., ischemic stroke), Alternatively, it can be used to improve something.

[0034] In specific embodiments, anti-FXI antibodies and / or anti-FXIa antibodies, or the same The antigen-binding fragment is determined, for example, by the aPTT assay described in the Examples section. In a concentration-dependent manner, the clotting time of human plasma (for example, the time until a blood clot begins to form) It is possible to extend it. In a specific embodiment, the clotting time (aPTT) is aPTT As determined by the assay, the range is 10 nM to 20 nM, for example, about 14 nM or Compared to baseline, with a 15 nM total anti-FXI antibody (e.g., NOV1401) concentration. It doubled. In certain embodiments, anti-FXI antibody and / or anti-FXIa antibody, These antigen-binding fragments are determined, for example, by the aPTT assay described in the Examples section. As described, in the range of 5nM to 20nM, for example, with an IC50 of approximately 13nM, the concentration is dependent. It is possible to prolong the clotting time of human plasma.

[0035] In specific embodiments, the anti-FXI antibody and / or anti-FXI described herein a antibody, or its antigen-binding fragment, is, for example, the aPTT app described in the Examples section. As determined by Sei, for example, in a concentration-dependent manner, at least 1.1 times, 1.2 times, 1 0.3x, 1.4x, 1.5x, 1.6x, 1.7x, 1.8x, 1.9x, or 2x, It is possible to extend the clotting time of human plasma (for example, the time it takes for a blood clot to begin forming). Yes. In specific embodiments, the anti-FXI antibody and / or anti-F described herein are used. XIa antibodies, or their antigen-binding fragments, are, for example, aPTT as described in the Examples section. As determined by the assay, at least 1.4 times, 1.5 times, 1.6 times, or 1. It is possible to extend the clotting time of human plasma (for example, the time until a blood clot begins to form) by 7 times. It is possible.

[0036] In specific embodiments, the anti-FXI antibody and / or anti-FXIa antibody described herein. The body, or its antigen-binding fragments, in the presence of very low concentrations of tissue factor (TF) Therefore, the effect of FXIa inhibition on the thrombin → FXIa feedforward loop is In the thrombin production assay (TGA) of human plasma, the amount of thrombin is measured. It is possible to reduce it in a concentration-dependent manner. In certain embodiments, the anti- FXI antibodies and / or anti-FXIa antibodies, or their antigen-binding fragments, are found in human blood. In a thrombin synthesis assay (TGA), in the range of 10 nM to 30 nM, for example ICs with a frequency of approximately 20nM or 24nM 50 With a residual thrombin concentration of approximately 159 nM, It is possible to reduce the amount of thrombin.

[0037] In specific embodiments of this specification, the catalytic domain of human FXI and / or FXIa is It specifically binds and the terminal phase elimination half-life (t) of all antibodies in cynomolgus monkeys. 1 / 2 ) to approximately 1 The incubation period is 4 to 15 days, and the antibody (e.g., NOV1401, or HCDR of NOV1401) is used. Antibodies including 1-3 and LCDR1-3 (as listed in Table 1), or their antigen binding. Sex fragments are presented. In specific embodiments, such anti-FXI / FXIa antibodies are approximately It exhibits absolute subcutaneous (sc) bioavailability of 61-66%.

[0038] In a specific embodiment, this specifically binds to human FXI and / or FXIa. The antibody or its antigen-binding fragment presented in the specification (e.g., NOV1401, see Table 1) The antibodies listed in [the document] have the following characteristics: (i) For example, apparent K at approximately 1-2 pM and 4-5 pM respectively D So, human F Specific binding to the catalytic domain (CD) of XI and FXIa; (ii) Condensation assessed by activated partial thromboplastin time (aPTT) assay Prolonging the blood flow time; (iii) F by activated factor XII (FXIIa) and thrombin, respectively Inhibiting thrombin production in human plasma by inhibiting XI activation; (iv) In FXI- / - mice reconstituted with human FXI, antithrombotic activity and anti To exhibit coagulation activity; (v) For example, in cynomolgus monkeys, free FXI (FXI f ) Reduce the level, Or extend the reduction; (vi) For example, the terminal phase elimination half-life of all antibodies in cynomolgus monkeys is approximately 14-15 days. To make a space; (vii) Although it specifically binds to human and monkey FXI and / or FXIa, It does not specifically bind to mouse or rat FXI and / or FXIa; Bini (viii) The following residues of human FXI (numbered by Swissprot): Pro 410, Arg413, Leu415, Cys416, His431, Cys432, T yr434, Gly435, Glu437, Tyr472~Glu476, Tyr521 ~Lys527, Arg548, His552, Ser575, Ser594~Glu5 97, and one or more of Arg602~Arg604 (for example, two, three) (4, 5, 6, or 7, or more), or some or all of them Contact One or more of these (for example, two, or three, or four, or five, or It presents six, seven, or all of them.

[0039] The isolated anti-FXI antibody and / or isolated FXIa antibody described herein, or The antigen-binding fragments include monoclonal antibodies, human or humanized antibodies, chimeric antibodies, and single-chain antibodies. Body, Fab fragment, Fv fragment, F(ab')2 fragment, or scFv fragment, and / or It could be an IgG isotype (for example, IgG1 such as human IgG1). In the application form, the anti-FXI antibody and / or anti-FXIa antibody described herein is recombinant It is a human antibody. In specific embodiments, the anti-FXI antibody and / Alternatively, the anti-FXIa antibody is a human IgG1 / lambda (λ) antibody. In specific embodiments... The anti-FXI antibodies and / or anti-FXIa antibodies described herein are effectors. F manipulated to reduce the likelihood of function (e.g., ADCC and / or CDC) Human Fc domains containing c domains, e.g., D265A substitution and / or P329A substitution. This is a human IgG1 / lambda (λ) antibody containing yin.

[0040] The isolated anti-FXI antibody and / or isolated FXIa antibody described herein or the Their antigen-binding fragments also contain amino acids that match human VH germline sequences or human VL germline sequences. Frameworks that have been replaced with antibody frameworks derived from each cell lineage sequence It may include.

[0041] Another aspect of the present invention has the complete heavy and light chain sequences of Fab as described in Table 1. It contains an isolated antibody or its antigen-binding fragment. More specifically, it contains an isolated antibody or its antigen-binding fragment. The compatible fragment may have the heavy and light chain sequences of NOV1090 and NOV1401.

[0042] Further aspects of the present invention include the heavy and light chain variable domains of Fab as described in Table 1. It includes an isolated antibody having a sequence or an antigen-binding fragment thereof. More specifically, an isolated antibody or Its antigen-binding fragments are variable heavy and light chains of NOV1090 and NOV1401. It may have a main sequence.

[0043] Further aspects of the present invention include Kabat CDR, IMGT CDR, and Chothia The heavy chain variable domain CD of the antibodies listed in Table 1, such as CDR or combined CDR. R (i.e., HCDR1, HCDR2, and HCDR3) sequences and light chain variable domains Isolated CDR (i.e., LCDR1, LCDR2, and LCDR3) sequences It includes an antibody or its antigen-binding fragment. More specifically, an isolated antibody or its antigen-binding fragment Fragments are available as Kabat CDR, IMGT CDR, Chothia CDR, or combinations. For example, CDRs, such as NOV1090 and NOV1401, as presented in Table 1, H CDR1 sequence, HCDR2 sequence, HCDR3 sequence, LCDR1 sequence, LCDR2 sequence, It may also have an LCDR3 sequence.

[0044] The present invention also includes heavy chain CDR1 selected from the group consisting of sequence numbers 3 and 23, sequence number Heavy chain CDR2 selected from the group consisting of numbers 4 and 24, and sequence numbers 5 and 25 It contains a heavy chain CDR3 selected from the group consisting of human FXI and / or human FXI The invention also relates to an isolated antibody that binds to a or its antigen-binding fragment. In another embodiment, such The isolated antibody or its antigen-binding fragment was selected from the group consisting of SEQ ID NOs: 13 and 33. Light chain CDR1, light chain CDR2 selected from the group consisting of sequence numbers 14 and 34, The further comprises a light chain CDR3 selected from the group consisting of sequence numbers 15 and 35.

[0045] The present invention also includes a light chain CDR1 selected from the group consisting of SEQ ID NOs: 13 and 33, sequence Light chain CDR2 selected from the group consisting of numbers 14 and 34, and sequence number 15 and Includes a light chain CDR3 selected from the group consisting of 35, and human FXI and / or human This also relates to isolated antibodies that bind to FXIa or their antigen-binding fragments.

[0046] The present invention also presents isolated antibodies or antigenic antibodies that bind to FXI and / or FXIa. Composite fragments, including HCDR1, HCDR2, and HCDR3, as well as LCDR1, It has LCDR2 and LCDR3, and HCDR1, HCDR2, and HCDR3, Includes sequence numbers 3, 4, and 5, with LCDR1, LCDR2, and LCDR3 being sequence number 1 3, 14, and 15 are included, or HCDR1, HCDR2, and HCDR3 are Includes sequence numbers 23, 24, and 25, with LCDR1, LCDR2, and LCDR3 being sequence numbers. This also relates to isolated antibodies or their antigen-binding fragments, including those numbered 33, 34, and 35.

[0047] The present invention also presents isolated antibodies that bind to FXI and / or FXIa, or their antigenic binding. Sexual fragments, including HCDR1, HCDR2, and HCDR3, as well as LCDR1, L It has CDR2 and LCDR3, and HCDR1, HCDR2, and HCDR3 are Each includes sequence numbers 43, 44, and 45, respectively, and LCDR1, LCDR2, LCDR3 However, isolated antibodies containing SEQ ID NOs. 47, 37, and 15, respectively, or their antigen-binding decomposition, are included. It also concerns one side.

[0048] The present invention also presents isolated antibodies that bind to FXI and / or FXIa, or their antigenic binding. Sexual fragments, including HCDR1, HCDR2, and HCDR3, as well as LCDR1, L It has CDR2 and LCDR3, and HCDR1, HCDR2, and HCDR3 are Each includes sequence numbers 46, 4, and 5, and LCDR1, LCDR2, and LCDR3 are, Each of these isolates an antibody or its antigen-binding fragment containing SEQ ID NOs. 33, 14, and 15. It also relates to this.

[0049] The present invention also relates to the variable heavy chain of SEQ ID NOs. 9 and 29 as defined by Chothia. Variable light HCDR1, HCDR2, and HCDR3, as well as sequence numbers 19 and 39 This also relates to antibody or antigen-binding fragments having LCDR1, LCDR2, and LCDR3 in the chain. In another aspect of the present invention, the antibody or antigen-binding fragment is defined by Kabat. , the heavy chain variable domain sequences of SEQ ID NOs. 9 and 29 are HCDR1, HCDR2, and HC DR3, and LCDR1, LCD, the light chain variable domain sequences of sequence numbers 19 and 39. It may have R2 and LCDR3.

[0050] The present invention also relates to the variable heavy chain HCD of sequence numbers 9 and 29 as defined by IMGT. R1, HCDR2, and HCDR3, as well as the variable light chain L of sequence numbers 19 and 39. This also relates to antibodies or antigen-binding fragments having CDR1, LCDR2, and LCDR3. In another aspect of the present invention, the antibody or antigen-binding fragment is defined by Combined The heavy chain variable domain sequences of sequence numbers 9 and 29, HCDR1, HCDR2, and HCDR3, and the light chain variable domain sequences of SEQ ID NOs. 19 and 39, LCDR1, L It may have CDR2 and LCDR3.

[0051] In one aspect of the present invention, the isolated antibody or its antigen-binding fragment is SEQ ID NOs. 9 and 29. It contains a heavy chain variable domain sequence selected from the following groups. The isolated antibody or antigen-binding fragment is It may further include a light chain variable domain sequence, in which case the heavy chain variable domain and The light chain variable domain, when combined with other elements, forms an antigen-binding site for FXIa. In particular, the light chain variable domain sequence can be selected from sequence numbers 19 and 39, and this In this case, the isolated antibody or its antigen-binding fragment binds to FXI and / or FXIa. To combine.

[0052] The present invention also provides a light chain variable domain selected from the group consisting of SEQ ID NOs: 19 and 39. An isolated antibody or its antigen that includes a column and binds to human FXI and / or human FXIa. This also relates to binding fragments. Isolated antibodies or antigen-binding fragments further modify the heavy chain variable domain sequence. It is possible to include in this, and in this case, the light chain variable domain and the heavy chain variable domain are combined. This then forms antigen-binding sites for FXI and / or FXIa.

[0053] In particular, isolated antibodies or antigen-binding fragments that bind to FXI and / or FXIa. These include sequences 9 and 19; or 19 and 39, respectively, representing heavy and light chains. It may have a variable-chain domain.

[0054] The present invention further includes sequences selected from the group consisting of sequence numbers 9 and 29 and at least 8 Heavy with sequence identity of 0%, 85%, 90%, 95%, 97%, 98%, or 99% The term also relates to an isolated antibody containing a chain variable domain or its antigen-binding fragment, in which case the antibody It binds to FXI and / or FXIa. In one embodiment, the isolated antibody or its antigen The binding fragment also has at least one sequence selected from the group consisting of sequence numbers 19 and 39. It has sequence identity of 80%, 85%, 90%, 95%, 97%, 98%, or 99%. This also includes a light chain variable domain. In a further aspect of the present invention, the isolated antibody or antigen-binding fragment is , as defined by Kabat and listed in Table 1, HCDR1, HCDR2, HCD It has R3, LCDR1, LCDR2, and LCDR3. In a specific embodiment, The release antibody or antigen-binding fragment is defined by Chothia, IMGT, or a combination thereof. The following are listed in Table 1: HCDR1, HCDR2, HCDR3, LCDR1, LCD It has R2 and LCDR3.

[0055] The present invention also includes sequences selected from the group consisting of sequence numbers 19 and 39 and at least 8 Light with sequence identity of 0%, 85%, 90%, 95%, 97%, 98%, or 99% The also relates to isolated antibodies having a chain variable domain or their antigen-binding fragments, in this case the antibody The body binds to FXI and / or FXIa.

[0056] In another aspect of the present invention, an isolated antibody or so that binds to FXI and / or FXIa. The antigen-binding fragment may have a heavy chain containing the sequence of SEQ ID NO: 11 or 31. The isolated antibody is Furthermore, when combined with the heavy chain, it forms antigenic bonds against human FXI and / or human FXIa. It may also include light chains that can form bonding sites. In particular, the light chains may include sequences 21 or 41. It may have columns. In particular, isolated antibodies that bind to FXI and / or FXIa or their anti- The original binding fragments contain sequences 11 and 21; or 31 and 41, respectively. It may have heavy and light chains.

[0057] The present invention further comprises sequences selected from the group consisting of sequence numbers 11 or 31 and at least This also applies to isolated antibodies containing heavy chains with 90% sequence identity or their antigen-binding fragments. In this case, the antibody binds to FXI and / or FXIa. In one embodiment, The release antibody or its antigen-binding fragment is also selected from the group consisting of SEQ ID NOs: 21 or 41. The sequence is at least 80%, 85%, 90%, 95%, 97%, 98%, or 99% This also includes light chains that have sequence identity.

[0058] The present invention further comprises sequences selected from the group consisting of sequence numbers 21 or 41 and less They all have 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity. The present invention also relates to an isolated antibody containing a light chain or its antigen-binding fragment thereof, in which case the antibody is F Joins to XI and / or FXIa.

[0059] The present invention also includes compositions comprising an isolated antibody or its antigen-binding fragment as described herein. This also relates to antibody compositions combined with pharmaceutically acceptable carriers. For example, the present invention relates to antibodies such as NOV1090 and NOV1401, as shown in Table 1. The present invention further comprises a pharmaceutical composition containing the antigen-binding fragment thereof. The present invention also includes the isolated anti- The present invention also relates to pharmaceutical compositions comprising two or more combinations of the body or its antigen-binding fragments. .

[0060] The present invention also encodes a variable heavy chain having sequences selected from sequence numbers 9 and 29. This also relates to isolated nucleic acid sequences. In particular, nucleic acids are selected from the group consisting of sequence numbers 10 and 30. The sequence is at least 80%, 85%, 90%, 95%, 97%, 98%, or 99%. It has % sequence identity. In a further aspect of the present invention, the sequence is sequence number 10 or 30 That is the case.

[0061] The present invention also codes a variable light chain having sequences selected from sequence numbers 20 and 40. This also relates to the isolated nucleic acid sequences. In particular, the nucleic acids are selected from the group consisting of SEQ ID NOs. 20 and 40. Selected sequences and at least 80%, 85%, 90%, 95%, 97%, 98%, or 9 It has 9% sequence identity. In a further aspect of the present invention, the sequences are sequence numbers 20 and 4 It is 0.

[0062] The present invention also includes sequences selected from the group consisting of sequence numbers 20 and 40 and at least 9 Contains a sequence encoding a polypeptide containing a light chain variable domain with 0% sequence identity. This also relates to isolated nucleic acids.

[0063] The present invention also includes a vector comprising one or more nucleic acid molecules described herein. This also relates to that.

[0064] The present invention also includes a recombinant DNA sequence encoding the heavy chain of the antibody described above, and the above The same also applies to isolated host cells containing a second recombinant DNA sequence encoding the light chain of the antibody, In this case, the DNA sequence is operably linked to a promoter and expressed in a host cell. It is possible to do so. The antibody is intended to be a human monoclonal antibody. Furthermore, the host cell is intended to be a non-human mammalian cell.

[0065] The present invention also relates to the expression of FXI and / or FXIa, and / or intrinsic coagulation. A method for reducing the activation of pathways and / or common coagulation pathways, wherein cells are subjected to an effective amount , contact with a composition containing an isolated antibody or its antigen-binding fragment as described herein. This also concerns methods that include steps.

[0066] The present invention also provides a method for inhibiting the coupling of FXI and / or FXIa to FIX. There, the cells are subjected to an effective amount of the isolated antibody or its antigen-binding fragment described herein. The invention also relates to a method which includes the step of bringing the composition into contact with the present material.

[0067] The cells are intended to be human cells. The cells are further intended to be present in the object. In one embodiment, the cells are intended to be platelets. The subject is human. This is being considered even more.

[0068] The present invention also relates to a method for treating, improving, or preventing thromboembolic disease in a subject. Then, an effective amount of a set comprising the antibody or its antigen-binding fragment described herein is applied to the target. The invention also relates to a method that includes the step of administering the product. In one embodiment, thromboembolic disease is a thrombus Sexual disorders (e.g., thrombosis, thrombotic stroke, atrial fibrillation, prevention of stroke in atrial fibrillation (S) These include PAF, deep vein thrombosis, venous thromboembolism, and pulmonary embolism. The subjects are humans. A certain plan is also being considered.

[0069] Any of the isolated antibodies or their antigen-binding fragments mentioned above are monoclonal antibodies. It may be an antigen-binding fragment thereof.

[0070] Non-limiting embodiments of this disclosure are described in the following ways: 1. Isolated anti-FXI antibodies that bind to the catalytic domain of FXI and / or FXIa, and / or isolated anti-FXIa antibodies or fragments thereof. 2. Isolated antibodies that conjugate to one or more epitopes of anti-FXI and / or FXIa. or a fragment thereof, the epitope being Pro410, Arg413, Leu415, Cys416, His431, Cys432, Tyr434, Gly435, Glu43 7, Tyr472, Lys473, Met474, Ala475, Glu476, Tyr 521, Arg522, Lys523, Leu524, Arg525, Asp526, L ys527, Arg548, His552, Ser575, Ser594, Trp595 Gly596, Glu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment thereof containing two or more amino acid residues. 3. Epitope is Pro410, Arg413, Leu415, Cys416, His 431, Cys432, Tyr434, Gly435, Glu437, Tyr472, L ys473, Met474, Ala475, Glu476, Tyr521, Arg522 , Lys523, Leu524, Arg525, Asp526, Lys527, Arg5 48, His552, Ser575, Ser594, Trp595, Gly596, Gl Four or more amino acids from u597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to embodiment 2, comprising an acid residue. 4. Epitope is Pro410, Arg413, Leu415, Cys416, His 431, Cys432, Tyr434, Gly435, Glu437, Tyr472, L ys473, Met474, Ala475, Glu476, Tyr521, Arg522 , Lys523, Leu524, Arg525, Asp526, Lys527, Arg5 48, His552, Ser575, Ser594, Trp595, Gly596, Gl Six or more amino acids from among u597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to embodiment 2, comprising an acid residue. 5. Epitope is Pro410, Arg413, Leu415, Cys416, His 431, Cys432, Tyr434, Gly435, Glu437, Tyr472, L ys473, Met474, Ala475, Glu476, Tyr521, Arg522 , Lys523, Leu524, Arg525, Asp526, Lys527, Arg5 48, His552, Ser575, Ser594, Trp595, Gly596, Gl Eight or more amino acids from among u597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to embodiment 2, comprising an acid residue. 6. Epitope is Pro410, Arg413, Leu415, Cys416, His 431, Cys432, Tyr434, Gly435, Glu437, Tyr472, L ys473, Met474, Ala475, Glu476, Tyr521, Arg522 , Lys523, Leu524, Arg525, Asp526, Lys527, Arg5 48, His552, Ser575, Ser594, Trp595, Gly596, Gl A single compound of embodiment 2, comprising residues u597, Arg602, Glu603, and Arg604. Antibody release or fragment. 7. The epitopes are the amino acid residues Pro410, Arg413, and Lys527, and Le u415, Cys416, His431, Cys432, Tyr434, Gly435, Glu437, Tyr472, Lys473, Met474, Ala475, Glu47 6, Tyr521, Arg522, Lys523, Leu524, Arg525, Asp 526, Arg548, His552, Ser575, Ser594, Trp595, G Of ly596, Glu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to embodiment 2, comprising one or more amino acid residues. 8. The epitopes are the amino acid residues Pro410, Arg413, and Lys527, and Le u415, Cys416, His431, Cys432, Tyr434, Gly435, Glu437, Tyr472, Lys473, Met474, Ala475, Glu47 6, Tyr521, Arg522, Lys523, Leu524, Arg525, Asp 526, Arg548, His552, Ser575, Ser594, Trp595, G Of ly596, Glu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to embodiment 2, comprising four or more amino acid residues. 9. The epitopes are the amino acid residues Pro410, Arg413, and Lys527, and Le u415, Cys416, His431, Cys432, Tyr434, Gly435, Glu437, Tyr472, Lys473, Met474, Ala475, Glu47 6, Tyr521, Arg522, Lys523, Leu524, Arg525, Asp 526, Arg548, His552, Ser575, Ser594, Trp595, G Of ly596, Glu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to embodiment 2, comprising six or more amino acid residues. 10. Isolated anti-FXI antibodies that bind to the catalytic domain of FXI and / or FXIa. and / or isolated anti-FXIa antibodies or fragments thereof, wherein FXI and / or F XIa to one or more of Factor IX, Factor XIIa, and thrombin An isolated antibody or fragment that blocks binding. 11. Factor IX, Factor XIIa, or thrombi of FXI and / or FXIa Aspect 10, which blocks binding to n and one or more of the other components of the coagulation pathway. Isolated antibody or fragment. 12. One or more of FIX, FXI, and FXIa binding to platelet receptors An isolated antibody or fragment according to embodiment 1 that blocks the compound. 13. An isolated antibody according to Embodiment 1 that prevents activation of the intrinsic coagulation pathway or the common coagulation pathway. piece. 14. In human FXI protein and / or human FXIa protein, BIACORE (Trademark) assays measuring 34 nM or less, or in a solution equilibrium titration assay (S K(s) less than 4 pM as measured by ET D An isolated antibody or fragment thereof that binds to it. 15. For at least one of the CDRs listed in Table 1, at least 90% of the same An isolated antibody or fragment according to Embodiment 1, comprising at least one complementarity-determining region having unisexuality. 16. Isolated antibody or fragment according to Embodiment 1, comprising CDR1, CDR2, and CDR3 of Table 1. . 17. Isolation variants of the antibody or fragment of Embodiment 1, wherein the antibody or fragment is C in Table 1. Includes DR1, CDR2, and CDR3, and the variant is CDR1, CDR2, or CD Isolated mutants having at least 1 to 4 amino acid changes in one of the R3 groups. . 18. A single unit of embodiment 1 comprising a heavy chain CDR3 selected from the group consisting of sequence numbers 5 and 25. Antibody release or fragment. 20. VH selected from the group consisting of SEQ ID NOs. 9 and 29, or 90% of the same. An amino acid sequence having the same identity; and selected from the group consisting of SEQ ID NOs: 19 and 39. A single amino acid sequence comprising a VL, or an amino acid sequence having 90% identity thereto, according to Embodiment 1 Antibody release or fragment. 21. VH selected from the group consisting of SEQ ID NOs. 9 and 29, or 95% of the same. An amino acid sequence having the same identity; and selected from the group consisting of SEQ ID NOs: 19 and 39. A single amino acid sequence comprising a VL, or an amino acid sequence having 95% identity thereto, according to Embodiment 1 Antibody release or fragment. 22. VH selected from the group consisting of sequence numbers 9 and 29, or 97% of the same. An amino acid sequence having the same identity; and selected from the group consisting of SEQ ID NOs: 19 and 39. A single amino acid sequence comprising a VL, or an amino acid sequence having 97% identity thereto, according to Embodiment 1 Antibody release or fragment. 23. A single variable heavy chain sequence selected from the group consisting of sequence numbers 9 and 29, according to embodiment 1. Antibody release or fragment. 24. A variable light chain sequence selected from the group consisting of sequence numbers 19 and 39, comprising Embodiment 1 Isolated antibody or fragment. 25. Variable heavy chains selected from the group consisting of SEQ ID NOs: 9 and 29; and SEQ ID NO: 19 An isolated antibody or fragment of Embodiment 1 comprising a variable light chain sequence selected from the group consisting of 39 and 39. . 26. Antibodies or fragments containing the variable heavy chain sequence of SEQ ID NO: 9 and the variable light chain sequence of SEQ ID NO: 19. Furthermore, antibodies or cleavage containing the variable heavy chain sequence of SEQ ID NO: 29 and the variable light chain sequence of SEQ ID NO: 39 An isolated antibody or fragment according to embodiment 1, selected from the group consisting of pieces. 27. CDR1 of the heavy chain variable region selected from the group consisting of SEQ ID NO: 46; from SEQ ID NO: 4 CDR2 is selected from the group consisting of 5; CDR3 is selected from the group consisting of 5; Sequence ID 33 The group consisting of CDR1 of the light chain variable region selected from the group and sequence number 14 is selected from the group A single CDR3 selected from the group consisting of CDR2 and SEQ ID NO: 15, according to Embodiment 1. Antibody release or fragment. 28. CDR1 of the heavy chain variable region selected from the group consisting of Sequence IDs 3 and 23; Sequence No. CDR2 selected from the group consisting of numbers 4 and 24; CDR2 selected from the group consisting of numbers 5 and 25 CDR3; CDR of the light chain variable region selected from the group consisting of sequence numbers 13 and 33 1; CDR2 selected from the group consisting of SEQ ID NOs: 14 and 34; and SEQ ID NO: 15 An isolated antibody or fragment according to embodiment 1, comprising CDR3 selected from the group consisting of and 35. 29. CDR1 of the heavy chain variable region selected from the group consisting of sequence numbers 6 and 26; sequence number CDR2 selected from the group consisting of numbers 7 and 27; selected from the group consisting of numbers 8 and 28 CDR3; CDR of the light chain variable region selected from the group consisting of sequence numbers 16 and 36 1; CDR2 selected from the group consisting of SEQ ID NOs. 17 and 37; and SEQ ID NO. 18 An isolated antibody or fragment according to embodiment 1, comprising CDR3 selected from the group consisting of 38. 30. CDR1 of the heavy chain variable region of SEQ ID NO: 3; CDR2 of the heavy chain variable region of SEQ ID NO: 4; distribution CDR3 of the heavy chain variable region in column number 5; CDR1 of the light chain variable region in sequence number 13; sequence number Including CDR2 of 14 light chain variable regions; and CDR3 of the light chain variable region of sequence number 15, An isolated antibody or fragment according to Embodiment 1. 31. CDR1 of the heavy chain variable region of SEQ ID NO: 23; CDR2 of the heavy chain variable region of SEQ ID NO: 24 ;CDR3 of the heavy chain variable region of SEQ ID NO: 25;CDR1 of the light chain variable region of SEQ ID NO: 33; CDR2 of the light chain variable region at column number 34; and CDR3 of the light chain variable region at sequence number 35. Including an isolated antibody or fragment of Embodiment 1. 32. CDR1 of the heavy chain variable region of SEQ ID NO: 6; CDR2 of the heavy chain variable region of SEQ ID NO: 7; distribution CDR3 of the heavy chain variable region in column number 8; CDR1 of the light chain variable region in sequence number 16; sequence number Including CDR2 of 17 light chain variable regions; and CDR3 of the light chain variable region of sequence number 18, An isolated antibody or fragment according to Embodiment 1. 33. CDR1 of the heavy chain variable region of SEQ ID NO: 26; CDR2 of the heavy chain variable region of SEQ ID NO: 27 ;CDR3 of the heavy chain variable region of SEQ ID NO: 28;CDR1 of the light chain variable region of SEQ ID NO: 36; CDR2 of the light chain variable region in column number 37; and CDR3 of the light chain variable region in sequence number 38 Including an isolated antibody or fragment of Embodiment 1. 34. A pharmaceutical compound comprising an antibody or fragment thereof according to one of the above embodiments and a pharmaceutically acceptable carrier. Finished product. 35. A state that binds to the same epitope as the isolated antibody or fragment described in any of the above embodiments. Isolated antibody or fragment of type 1. 36. Regarding binding to human FXI protein and / or human FXIa protein, The isolated antibody or fragment of Embodiment 1 competes with the isolated antibody or fragment of any of the embodiments described above. Piece. 37. Isolated antimicrobial agent of embodiment 1, selected from the group consisting of NOV1090 and NOV1401 A body or fragment. 38. A method for treating thromboembolic disorders, comprising an effective amount, as described in any of the above embodiments. A pharmaceutical composition containing an antibody or fragment is administered to a subject suffering from a thromboembolic disorder. A method that includes steps. 39. The subject has one of the following conditions associated with atrial fibrillation: ischemic stroke or deep vein thrombosis. The method of aspect 38 involves multiple illnesses. 40. The method of aspect 38, wherein the subject suffers from ischemic stroke associated with atrial fibrillation. 41. A method for treating thromboembolic disorders, comprising an effective amount, as described in any of the above embodiments. A pharmaceutical composition containing antibodies or fragments, in combination with statin therapy, is used to treat thromboembolic disorders. A method that includes the step of administering an agent to a patient. 42. A pharmaceutical product comprising the antibody described in any of the above embodiments. 43. A nucleic acid encoding one or more of the antibodies described in any of the above embodiments. 44. A vector containing nucleic acid as described in embodiment 43. 45. A host cell containing the vector described in embodiment 44. 46. ​​When it binds to the active FXI(FXIa) catalytic domain, it confuses FXIa. The motion is such that the 4 residues at the N-terminus, loops 145, 188, and 220 are shifted and / or irregular compared to the active conformation, and are changed to an inactive conformation. An isolated antibody or fragment of embodiment 1. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where the 4 residues at the N-terminus, loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. 47. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where the 4 residues at the N-terminus, loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. 48. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where the 4 residues at the N-terminus, loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where the 4 residues at the N-terminus, loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. 49. When binding to FXI, it induces a conformational change in the prothrombin structure, and further results in an inhibitory FXI conformation closely related to the inhibitory FXI conformation observed when binding to FXIa, thereby preventing the FXI catalytic domain from taking an active conformation. An isolated antibody or fragment of embodiment 1. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where the 4 residues at the N-terminus, loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. When binding to FXI, the FXI catalytic domain is prevented from taking an active conformation where the 4 residues at the N-terminus, loops 145, 188, and 220 are regular as in the structure of the FXIa catalytic domain. An isolated antibody or fragment of embodiment 1. 50. When binding to FXI and / or FXIa and forming an antibody:antigen complex with the catalytic domain of FXI and / or FXIa, it causes a shift and / or loss of orientation of loops 145, 188, and 220 compared to the uncomplexed structure of the catalytic domain of active factor XI (FXIa). An isolated antibody or fragment of embodiment 1. When binding to FXI and / or FXIa and forming an antibody:antigen complex with the catalytic domain of FXI and / or FXIa, it causes a shift and / or loss of orientation of loops 145, 188, and 220 compared to the uncomplexed structure of the catalytic domain of active factor XI (FXIa). An isolated antibody or fragment of embodiment 1. When binding to FXI and / or FXIa and forming an antibody:antigen complex with the catalytic domain of FXI and / or FXIa, it causes a shift and / or loss of orientation of loops 145, 188, and 220 compared to the uncomplexed structure of the catalytic domain of active factor XI (FXIa). An isolated antibody or fragment of embodiment 1. 51. When binding to FXI and / or FXIa and forming an antibody:antigen complex with the catalytic domain of FXI and / or FXIa, it causes a shift and / or loss of orientation of the 4 residues at the N-terminus, loops 145, 188, and 220 compared to the uncomplexed structure of the catalytic domain of active factor XI (FXIa). When binding to FXI and / or FXIa and forming an antibody:antigen complex with the catalytic domain of FXI and / or FXIa, it causes a shift and / or loss of orientation of the 4 residues at the N-terminus, loops 145, 188, and 220 compared to the uncomplexed structure of the catalytic domain of active factor XI (FXIa). Isolation of embodiment 1 that causes loss of shift and / or orientation when compared to the i structure Antibody or fragment. 52. Binds to active FXI (FXIa) and to the FXI (FXIa) catalytic domain, its co nformation, where loops 145, 188, and 220 are in an inactive conformation shifted compared to the active conformation and / or have lost their orientation and is converted to an inactive conformation Isolated antibody or fragment of embodiment 1.

[0071] Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0072] The terms "FXI protein", "FXI antigen", and "FXI" are used interchangeably and refer to factor XI proteins in different species. Factor XI is a zymogen that, upon limited proteolysis, is converted to an active serine protease and participates in the intrinsic pathway of blood coagulation as a glycoprotein present in human plasma at a concentration of 25 - 30 nM, and is the mammalian plasma coagulation factor XI.

[0073] The terms "FXIa protein", "FXIa antigen", and "FXIa" are used interchangeably and refer to activated FXI proteins in different species. Factor XI, a zymogen, is converted to its active form, factor Xla coagulation factor (FXIa), via the contact phase of blood coagulation or via thrombin-mediated activation on the platelet surface. Upon activation of factor XI, the internal peptide bonds are cleaved in each of the two chains and disulf ​​​​​A serine is composed of two heavy chains and two light chains, held together by phytobonds. This serine protease results in activated Xla factor, which is a rotease. XFIa converts coagulation factor IX to IXa, and then IXa converts to coagulation factor X. Xa activates coagulation factors. Subsequently, Xa activates the second coagulation factor / thrombin. It can mediate. For example, human FXI has the sequence shown in Table 1 (Sequence ID 1), and precedent Reports and literature (Mandle RJ Jr, et al. (1979) Blood; 54(4):850; NCBI reference sequence) This is described in :AAA51985).

[0074] In the context of this invention, the terms "FXI" and "FXIa" (etc.) refer to natural FX Mutants and variants of protein I and protein FXIa, respectively The amino acid sequence of the natural primary structure (amino acid sequence) described in the above report and This includes mutants and variants that have substantially the same amino acid sequence.

[0075] The terms "catalytic domain," "serine protease catalytic domain," and the foregoing are used herein. A similar term used is the number of Glu1s counted from the N-terminus of a circulating mature protein. This refers to the mino acid Ile370~Val607. This also refers to the residue at the C-terminus of FXI. It can also be described as bases 388-625. The term "active site" as used in this specification and The term refers to the amino acids His413, Asp462, and Se557, which are toxic This refers to the three pharmacokinetic residues (Bane and Gailani (2014) Drug Disc. 19(9)).

[0076] The term "about" with respect to a numerical value x means, for example, x ± 10%.

[0077] As used herein, the term "antibody" means an intact antibody and any antigen-binding fragment (i.e., an "antigen-binding portion") or single chains thereof. An intact antibody is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region comprises one domain, CL. The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). Each VH and each VL is composed of three CDRs and four FRs arranged in the following order from the amino terminus to the carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain the binding domains that interact with an antigen. The constant regions of the antibody mediate binding of the immunoglobulin to various cells of the host tissues or immune system (e.g., effector cells) and factors including the first component (Clq) of the classical complement system. As used herein, the term "antigen-binding portion" or "antigen-binding fragment" of an antibody refers to one or more fragments of an intact antibody that retain the ability to specifically bind to a given antigen (e.g., factor XIa (FXIa)). The antigen-binding function of an antibody can be

[0078] Even more preferably, the antibody is a monoclonal antibody, a human antibody, a humanized antibody, or a chimeric antibody. As used herein, the term "antigen-binding portion" or "antigen-binding fragment" of an antibody refers to one or more fragments of an intact antibody that retain the ability to specifically bind to a given antigen (e.g., factor XIa (FXIa)). ​​​​​​​​​​​​ This can be accomplished by a fragment of an antibody. The antigen-binding portion or antigen-binding fragment of the antibody Examples of binding fragments included within the term are Fab fragments, VL domains, VH domains, and C A monovalent fragment consisting of an L domain and a CH1 domain; disulfide in the hinge region F(ab)2 fragment is a divalent fragment containing two Fab fragments linked by crosslinking; VH Fd fragment consisting of the main and CH1 domains; VL domain of a single arm of the antibody Fv fragment consisting of VH domains; single domain consisting of VH domain or VL domain Antibody (dAb) fragments (Ward et al., 1989 Nature 341:544~546); and isolation and complementarity determination. Includes area (CDR).

[0079] Furthermore, the two domains of the Fv fragment, the VL domain and the VH domain, are separate Encoded by genes, but using recombination, they can be processed into a single protein chain. It can be attached by an artificial peptide linker that makes it possible to produce, in this case The VL and VH regions pair up to form a monovalent molecule (known as single-chain Fv(scFv)). For example, Bird et al., 1988 Science 242:423~426; and Huston et al., 1988 Proc. Natl. See Acad. Sci. 85:5879~5883. Such single-chain antibodies form antibodies It contains one or more antigen-binding moieties or antigen-binding fragments. These antibody fragments Obtained using conventional techniques known to the manufacturer, the fragments are also useful in the same form as intact antibodies. Screening will be conducted for this.

[0080] Antigen-binding fragments also include single-domain antibodies, maxi-bodies, mini-bodies, and intra-bodies. i, Diabody, Triabody, Tetrabody, v-NAR, and bis-scFv (For example, Hollinger and Hudson, 2005, Nature Biotechnology, 23, 9, 1126-1136) It can also be incorporated into (see reference). The antigen-binding portion of the antibody is type III fibrous. Polypeptides such as nectin (Fn3) (fibronectin polypeptide monobody) To scaffolding based on U.S. Patent No. 6,703,199 (see U.S. Patent No. 6,703,199 for further details) It can be grafted.

[0081] The antigen-binding fragment, together with a complementary light chain polypeptide, forms a pair of antigen-binding domains. It is incorporated into a single-chain molecule containing a pair of tandem Fv segments (VH-CH1-VH-CH1). It can be incorporated (Zapata et al., 1995 Protein Eng. 8(10):1057~1062; and US Patent No. 5,641,870).

[0082] As used herein, the term "affinity" refers to the resistance of a single antigenic site. This refers to the strength of the interaction between the body and the antigen. Within each antigenic site, the variable region of the antibody "arm" The region interacts with the antigen via weak non-covalent forces at numerous sites, and the interaction The larger the size, the stronger the affinity. Antibodies or their antigen-binding fragments (e.g., Fa With respect to fragment b), the term “high affinity” as used herein generally refers to 10 -9 K below M D (For example, 10 -10 K below M D , 10 -11 K below M D , 10 - 12 K below MD , 10 -13 K below M D , 10 -14 K below M D Anti- Refers to a body or antigen-binding fragment.

[0083] The term "amino acid" includes naturally occurring amino acids and synthetic amino acids, as well as naturally occurring amino acids. This also refers to amino acid analogs and amino acid mimes that function in a similar manner to amino acids. The amino acids in question include not only those encoded by the genetic code, but also those that have been modified later. No acids, for example, hydroxyproline, γ-carboxyglutamic acid, and O-phosphosate. It is also phosphorus. Amino acid analogs are compounds that have the same basic chemical structure as naturally occurring amino acids, in other words, Hydrogen, carboxyl group, amino group, and R group, for example, homoserine, norleucine , methionine sulfoxide, methionine methylsulfonium has an alpha carbon bonded to it This refers to compounds that have a modified R group (e.g., norleucine) or modified Although it has a peptide skeleton, it retains the same basic chemical structure as naturally occurring amino acids. Acid mimetic compounds are compounds that have a structure different from the typical chemical structure of amino acids, but are not naturally occurring amino acids. This refers to chemical compounds that function in a similar manner to acids.

[0084] As used herein, the term “binding specificity” refers to the reaction with only one antigenic determinant. This refers to the ability of individual antigen-binding sites.

[0085] Antibodies (e.g., FXI-binding antibodies and / or FXIa-binding antibodies) are "specifically" The phrase "to bind (or selectively)" refers to a heterogeneous group of proteins and other biomolecules. Cognitive antigens within (e.g., human FXI and / or FXIa or cynoquiza) This refers to the binding reaction that determines the presence of FXI and / or FXIa. The phrases "antibody that recognizes an antigen" and "antibody that is specific to an antigen" are equivalent to "antibody that is specific to an antigen." This term is used interchangeably with "antibody that binds to."

[0086] The term "mediating FXI and / or FXIa" means FXI and / or F XIa is the IXth factor (also known as FIX), the Xth factor (FX), and / or by directly or indirectly activating thrombin, and / also By binding to platelet receptors, it mediates the intrinsic coagulation pathway and / or common coagulation pathway. It refers to the fact of mediation.

[0087] The term "hemostasis" refers to the primary mechanism for stopping blood flow at the site of injury, and wound This represents each of the major mechanisms for restoring vascular patency during healing. Normal hemostasis and disease In physical thrombosis, there are three mechanisms: primary thrombosis, which involves the interaction of activated platelets with the blood vessel wall. The formation of blood, fibrin, and a process called fibrinolysis are activated simultaneously.

[0088] Terms such as "coagulation and coagulation cascade" and "coagulation cascade model" are used in relation to wounds. A protein-based system used to stabilize the blood clot that forms to seal the blood vessel. This refers to the coagulation pathway, which is a proteolytic cascade. Each enzyme in the pathway is present in the plasma. When activated, it undergoes proteolytic cleavage, releasing the activating factor from the precursor molecule. It exists as zymogen (in an inactive form). The coagulation cascade controls the activation process. It functions as a series of positive and negative feedback loops that control the process. The ultimate goal of the process is to Then, thrombin can convert soluble fibrinogen into fibrin, which forms blood clots. The purpose is to produce [something].

[0089] The process of thrombin production involves three phases: the intrinsic pathway, and the activation of the coagulation factor FXa (active The extrinsic pathway, an alternative pathway for generating sexualized factor X, and thrombin It can be divided into a final common path that results in the formation of (Hoffman MM a nd Monroe DM (2005) Curr Hematol Rep. 4:391 -396; Johne J, et al. (2006) Biol Chem. 387:173-178).

[0090] "Platelet aggregation" refers to the phenomenon that occurs when a blood vessel ruptures, where substances that normally do not come into direct contact with the blood flow... This refers to the process of exposure to these substances (mainly collagen and von Willebrand oxalool). The platelet allows the platelet to adhere to the fractured surface. Then, it releases chemicals that attract more platelets to the damaged area, but this is called platelet aggregation. These two processes are the initial response to stop bleeding.

[0091] As used herein, the term "thromboembolic disorder" or similar terms refers to intrinsic coagulation. The pathway and / or common coagulation pathway are abnormally activated or spontaneously inactivated. (For example, without treatment) refers to any number of conditions or diseases. These conditions are Thrombotic stroke, atrial fibrillation, prevention of stroke in atrial fibrillation (SPAF), deep vein thrombosis This includes, but is not limited to, venous thromboembolism and pulmonary embolism. These also include, Catheter-related conditions in which catheters become thrombotic (e.g., Hickman catheters in cancer patients) This includes extracorporeal membrane oxygenation (ECMO), where the tubing causes blood clotting. Shut up.

[0092] As used herein, the term "thromboembolic" or similar terms also refers to the anti-inhibitory properties of the present invention. Using FXI Ab and / or anti-FXIa Ab or their antigen-binding fragments The following can be prevented or mitigated: Paroxysmal atrial fibrillation or paroxysmal atrial flutter, persistent atrial fibrillation or persistent atrial flutter, Alternatively, if cardiac arrhythmias such as persistent atrial fibrillation or persistent atrial flutter are suspected or confirmed. Thromboembolism in the subject being treated; • Subjects with atrial fibrillation requiring stroke prevention (SPAF), and whose subgroups are The target group consists of AF patients undergoing percutaneous coronary intervention (PCI); • Management of acute venous thromboembolic events (VTE) in patients at high risk of bleeding. and prevention of long-term secondary VTE; • In secondary prevention following transient ischemic attack (TIA) or non-functional stroke For the prevention of thromboembolic events in cerebral and cardiovascular events, as well as in heart failure with sinus rhythm. Cerebral and cardiovascular events in this context; • Clot formation and thromboembolism in the left atrium in patients undergoing cardiac defibrillation for cardiac arrhythmias. Embolism; Thrombosis before, during, and after ablation procedures for cardiac arrhythmias; • Venous thrombosis, which is deep vein thrombosis or superficial vein thrombosis in the lower or upper extremities. Vessel thrombosis, thrombosis in the abdominal and thoracic veins, sinus thrombosis and jugular vein thrombosis This includes, but is not limited to, treatment and secondary prevention; Thrombosis on any artificial surface within a vein, such as a catheter or pacemaker lead; • Pulmonary embolism in patients with or without venous thrombosis; Chronic thromboembolic pulmonary hypertension (CTEPH); • Arterial thrombosis on ruptured atherosclerotic plaque, on arterial prostheses or catheters These are thrombosis in the arteries and thrombosis in seemingly normal arteries, and these are acute coronary syndrome. Group, ST-elevation myocardial infarction, non-ST-elevation myocardial infarction, unstable angina, stent thrombosis, arterial Thrombosis of any artificial surface within the system, and in subjects with or without pulmonary hypertension. This includes, but is not limited to, pulmonary thrombosis; • Thrombosis and thrombosis in patients undergoing percutaneous coronary intervention (PCI) Embolism; • Cardioembolic stroke and cryptogenic stroke; Thrombosis in patients with invasive and non-invasive cancerous malignancies; • Thrombosis involving indwelling catheters; • Thrombosis and thromboembolism in critically ill patients; • Cardiac thrombosis and thromboembolism, including cardiac thrombosis after myocardial infarction and cardiovascular thrombosis. Conditions such as aneurysms, cardiomyopathy, cardiac hypertrophy and cardiac dysfunction, myocarditis, and artificial surfaces within the heart This includes, but is not limited to, cardiothrombosis associated with the condition; Thromboembolism in patients with valvular heart disease, with or without atrial fibrillation; • Thromboembolism involving mechanical or biological prostheses for heart valves; • After cardiac repair for simple or complex cardiac malformations, natural or artificial cardiac pads Thromboembolism in patients with arterial or venous conduits; • Total knee replacement, total hip replacement, and orthopedic surgery, thoracic surgery, or abdominal surgery Venous thrombosis and thromboembolism following; • Following neurosurgical procedures including intracranial and spinal interventions Arterial thrombosis or venous thrombosis; • Factor V Leiden, prothrombin mutation, antithrombin III, prote Protein C deficiency and protein S deficiency, factor XIII mutations, familial fibrosis Linogenemia, congenital plasminogen deficiency, elevated factor XI levels, sickle cell disease, Antiphospholipid syndrome, autoimmune diseases, chronic bowel disease, nephrotic syndrome, hemolytic uremic disease, bone marrow Proliferative disorders, disseminated intravascular coagulation, paroxysmal nocturnal hemoglobinuria, and heparin-induced hematologic dysplasia Congenital or acquired thrombotic predisposition, including but not limited to thrombotic plaque; • Thrombosis and thromboembolism in chronic kidney disease; and • Thrombosis and Thromboembolism It may also refer to any number among them.

[0093] The term "chimeric antibody" refers to (a) the antigen-binding site (variable region), class, effervescence. The effector function, and / or the type, or class, the effector function, and / or a modified constant region of the species, or a completely different one that confers new properties to a chimeric antibody. To link to molecules such as enzymes, toxins, hormones, growth factors, drugs, etc. Antibody molecules in which the normal region or a part thereof has been modified, replaced, or exchanged; or (b) The variable region or a part thereof has different antigen specificity or altered antigen specificity. This refers to antibody molecules that have been modified, replaced, or exchanged using a specified variable region. For example, mouse antibodies use a constant region derived from human immunoglobulins. It can be modified by changing. Due to replacement by the human constant region, texture The antibody reduces antigenicity in humans compared to the original mouse antibody, while still recognizing the antigen. It can retain its unique characteristics in that context.

[0094] The term "conservatively modified variant" refers to a variant of either the amino acid sequence or the nucleic acid sequence. This also applies. With respect to a specific nucleic acid sequence, a conservatively modified variant is the same as an amino acid. Nucleic acids that encode an amino acid sequence or an essentially identical amino acid sequence, or nucleic acids that encode an amino acid sequence If a sequence does not code, it refers to essentially the same sequence. Due to the degenerate nature of genetic coding, A large number of functionally identical nucleic acids code for any given protein. For example, codons GCA, GCC, GCG, and GCU all contain the amino acid alanine. Therefore, alanine is coded at every position specified by the codon. Without modifying the polypeptide, the codon is changed to one of the corresponding codons listed. It can be modified. Such nucleic acid mutations are one type of conservatively modified mutation. It is a "silent mutation." Any nucleic acid encoding polypeptide as defined herein The column also describes all possible silent mutations in nucleic acids. Those skilled in the art will know that within nucleic acids... Each codon (AUG, which is usually the codon for methionine only, and AUG, which is usually the codon for tryptophan only) It has been recognized that modifying the codon (excluding TGG) can result in a functionally identical molecule. They will recognize. Therefore, within each sequence described, nucleic acids encoding polypeptides Each of these silent mutations is implied.

[0095] In polypeptide sequences, "conservatively modified variants" are chemically similar to amino acids. Individual substitutions and deletions in polypeptide sequences, resulting in amino acid substitutions. , or addition. In the art, this includes conservative substitutions that present functionally similar amino acids. The table is well known. Such conservatively modified mutants are the polymorphic mutants and interspecific phases of the present invention. This adds to the same organism and alleles, and does not exclude them. The eight groups are: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamate 1) Asparagine (E); 2) Asparagine (N), Glutamine (Q); 3) Arginine (R), Li Syn(K); 5) Isoleucine(I), Leucine(L), Methionine(M), Valine(V) );6) Phenylalanine (F), tyrosine (Y), tryptophan (W);7) Serine (S), threonine (T); and (8) cysteine ​​(C), methionine (M) are related to each other. In contrast, it contains amino acids that are conservative substitutions (for example, Creighton, Proteins (1984) (See reference). In some embodiments, the term “conservative sequence modification” refers to an amino acid It does not significantly affect or modify the binding characteristics of antibodies containing the sequence. It is used to refer to amino acid modifications.

[0096] The term "epitope" refers to a protein determinant that is capable of specific binding to an antibody. Epitopes are typically composed of chemically active surface molecules, such as amino acids or sugar side chains. In addition to their distinctive three-dimensional structural features, they typically also possess distinctive charge characteristics. The difference between conformal epitopes and non-conformal epitopes is that the former binds to a denaturing solvent. It is distinguished in that it is lost in the presence of the latter, but the bond to the latter is not lost. In a test, one antibody is converted to a second antibody by any method well known to those skilled in the art. When two antibodies are shown to bind to the same epitope, they are said to "compete" with each other.

[0097] As used herein, the term "human antibody" refers to the framework region and the CDR region. Each region is intended to contain antibodies that have variable regions derived from human sequences. Furthermore, if the antibody contains a constant region, the constant region also contains such human sequences, for example If so, by human germline sequences or mutated versions of human germline sequences The present invention relates to human antibodies that have amino acid residues not encoded by human sequences (for example, i Induced by random mutagenesis or site-directed mutagenesis in the in vitro. A mutation introduced by a somatic mutation in vivo. (May include mutations.)

[0098] The term "human monoclonal antibody" refers to an antibody that exhibits a single binding specificity. Both the framework region and the CDR region contain variable regions derived from human sequences. This refers to an antibody that performs human immunoglobulin genetic testing. In one embodiment, a human monoclonal antibody is a human immunoglobulin gene. Prepare the child library using a phage display method for screening. ru.

[0099] "Humanized" antibodies are those that have low immunogenicity in humans but retain the reactivity of non-human antibodies. This is an antibody that, for example, retains the non-human CDR region and the rest of the antibody. In their human counterparts (i.e., the steady-state region and framework portion of the variable region), This can be achieved by doing so. For example, Morrison et al., Proc. Natl. Acad. Sci. U SA, 81:6851~6855, 1984; Morrison and Oi, Adv. Immunol., 44:65~92, 1988; Verho eyen et al., Science, 239:1534~1536, 1988; Padlan, Molec. Immun., 28:489~498, 1991 See also Padlan, Molec. Immun., 31:169-217, 1994. In addition to human manipulation techniques. Examples include the Xoma technology disclosed in US5,766,886, but these include Not limited.

[0100] "Identical" or "identical" in the context of two or more nucleic acid sequences or polypeptide sequences. The term "percent" refers to two or more sequences or subsequences being identical. Use one of the array comparison algorithms below, or perform manual alignment and The maximum correspondence is compared across a comparison range or designated area measured by visual inspection. If sequenced, the specified percentage of amino acid residues or nucleos of the two sequences. If the chid is the same (i.e., across the specified area, or if not specified) , 60% identity across the entire sequence, 65%, 70%, 75%, 80% at will, If the two sequences have 85%, 90%, 95%, or 99% identity, then they are "substantially" identical. They are "identical". By choice, identity is defined as having at least approximately 50 nucleotides (or 10 nucleotides). over a range of lengths of (no acid), or more preferably 100 to 500, or 1000 Over a region of length greater than a nucleotide (or 20, 50, or 200 amino acids or more) It exists.

[0101] In sequence comparison, one sequence acts as a reference sequence against which the test sequences are compared. This is typical. When using a sequence comparison algorithm, the sample sequence and the reference sequence are Input into the computer, specify sub-array coordinates if necessary, and then input the array algorithm program. Specify the Gram parameters. You can also use the default program parameters. Alternatively, you can also specify alternative parameters. Then, using the array comparison algorithm, Based on the program parameters, the sequence identity parity of the test sequence is compared with the reference sequence. Calculate the value.

[0102] As used herein, the "comparison region" is the region after the two sequences have been optimally sequenced. A reference sequence that can be compared with the same number of consecutive positions, ranging from 20 to 600, typically about 50 to 20. 0, more typically, a number bag with consecutive positions selected from a group consisting of approximately 100 to 150 numbers. Includes a reference to the segment by any one of the following. In this technical field, for comparison Sequence alignment methods are well known. The optimal sequence alignment for comparison is, for example, Sm The local homology algorithm by ith and Waterman (1970) Adv. Appl. Math. 2:482c This can also be done by homology according to Needleman and Wunsch, J. Mol. Biol. 48:443, 1970. This can also be done using a sex alignment algorithm, as demonstrated by Pearson and Lipman, and Proc. Nat'l. This can also be done using the similarity search method according to Acad. Sci. USA 85:2444, 1988, and these algorithms Computerized implementation of Gorhythm (Wisconsin Genetics Software) tware Package, Genetics Computer Group, 57 5 Science Dr., Madison, WI, GAP, BESTFIT, This can also be done by FASTA and TFASTA, and manual alignment and Visual inspection (e.g., Brent et al., Current Protocols in Molecular Biology, John Wiley & Sons) This can also be done by (see Ringbou, Inc., ed., 2003).

[0103] Algorithms suitable for determining sequence identity percentage and sequence similarity percentage The two examples are, respectively, Altschul et al., (1977) Nuc. Acids Res. 25:3389~3402; and As described by Altschul et al. (1990) J. Mol. Biol. 215:403~410, BLA The ST algorithm and the BLAST 2.0 algorithm are used. BLAST analysis is performed. The software for this purpose is from the National Center for Biotech This algorithm is published by Nology Information. When determining the sequence using words of the same length within a database array, a positive threshold score T is used. To identify short word lengths W in the query array that match or satisfy this condition This involves first identifying high-scoring sequence pairs (HSPs). T is the adjacent word score threshold. These are called values ​​(Altschul et al., previously cited). These initial adjacent word hits contain them It acts as a seed to initiate a search for HSPs with longer durations. Cumulative alignments As long as the core can be increased, word hits can be extended in any direction along each sequence. The cumulative score is calculated using the parameter M (matching residue pairs) in nucleotide sequences. Reward score (always > 0) and N (penalty for mismatched residues) The core (always < 0) is used for calculation. For amino acid sequences, the scoring matrix is ​​used. Use this to calculate the cumulative score. The cumulative alignment score is the maximum value achieved. If the amount X decreases; due to the accumulation of alignment of one or more negative score residues , if the cumulative score falls to zero or below; or if it reaches any end of the sequence This stops the expansion of word hits in each direction. (BLAST algorithm parameter) The W, T, and X values ​​determine the sensitivity and speed of the alignment. BLASTN In the program (for nucleotide sequences), the word length (W) is 11, and the expected value (E) is 10. ), using M=5 and N=-4 as defaults, compare both chains. Amino acid sequence The BLASTP program for this has a word length of 3 and an expected value (E) of 10, and 50 BLOSUM62 scoring matrices (Henikoff and Henikoff, Proc. See Natl. Acad. Sci. USA 89:10915, 1989) Alignment (B), 10 periods We will use the default values ​​(E), M=5, and N=-4 to compare both chains.

[0104] The BLAST algorithm also performs statistical analysis of the similarity between two sequences (e.g.) For example, see Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873~5787, 1993. (to be done) will also be implemented. One measure of similarity provided by the BLAST algorithm is The probability that a match occurs by chance between two nucleotide sequences or amino acid sequences. This is the minimum summation probability (P(N)) that yields the target. For example, comparing the test nucleic acid to a reference nucleic acid... The minimum sum probability when compared is less than approximately 0.2, and more preferably less than approximately 0.01. Yes, and most preferably, if the nucleic acid is less than about 0.001, it is considered similar to the reference sequence. It can be done.

[0105] The percentage of identity between two amino acid sequences is also expressed in the PAM120 weighted residue table, 1 ALIGN uses a gap length penalty of 2 and a gap penalty of 4. E. Meyers and W. Miller (Computer) were incorporated into the program (version 2.0). It can also be determined using the algorithm described in Appl. Biosci., 4:11~17, 1988. In addition, the percentage of identity between the two amino acid sequences is Blossom 62 matrix. Rix or PAM250 matrix, and 16, 14, 12, 10, 8, 6, Alternatively, use a gap weighting of 4 and a length weighting of 1, 2, 3, 4, 5, or 6. The GCG software package (available on the internet at gcg.com) Needleman and Wunsch (J. Mol, Biol. 48:44) were incorporated into the internal GAP program. The decision can also be made using the algorithm described in 4-453 (1970).

[0106] In addition to the sequence identity percentage mentioned above, two nucleic acid sequences or polypeptides are essentially Another indicator of identity is encoded by the first nucleic acid, as described below. The polypeptide is immunologically opposed to the polypeptide encoded by the second nucleic acid. It is cross-reactivity. Therefore, for example, if two peptides are only conserved substitutions Therefore, if they are different, the polypeptide is typically substantially identical to the second polypeptide. Another indicator that two nucleic acid sequences are substantially identical is described below. Two molecules or their complements hybridize with each other under strict conditions. Another indicator that the two nucleic acid sequences are substantially identical is that they use the same primer. The ability to amplify the sequence using this method.

[0107] The term "isolated antibody" refers to an antibody that substantially does not contain other antibodies with different antigen specificities. Refers to the body (for example, an isolated antibody that specifically binds to FXI and / or FXIa is F (Substantially contains antibodies that specifically bind to antigens other than XI and / or FXIa.) However, isolated antibodies that specifically bind to FXI and / or FXIa may not bind to other antigens. It may exhibit cross-reactivity with other intracellular substances and / or chemicals. Furthermore, isolated antibodies may react with other intracellular substances and / or chemicals. In some cases, the substance may be virtually nonexistent.

[0108] The term "isotype" refers to the antibody class resulting from heavy chain constant region genes. For example, it refers to IgM, IgE, IgG1, or IgG4 (IgG). Isotype It also includes a qualified version of one of these classes, whose qualification is the Fc function. Modify it, for example, by enhancing or reducing its effector function or binding to Fc receptors. It is applied to the sea urchin.

[0109] The term "K" used in this specification assoc " or "K a The term "antibody" refers to a specific antibody-antibody. While "K" is intended to refer to the association rate of inter-initiated interactions, as used herein, di s " or "K d The term refers to the dissociation rate of a particular antibody-antigen interaction. Intend to. The term "K" used herein D The term "K" is d K a Ratio to (that is, K d / K a This refers to the dissociation constant obtained from ) and is expressed as molar concentration (M). . Antibody K D The value can be determined using methods that are well established in the art. . Antibody K D Methods for determining this include the BIACORE(trademark) system and other biosensors. Surface plasmon resonance can be measured using a sensor system, or by solution equilibrium titration (SE). The procedure includes the step of measuring affinity in the solution by T).

[0110] The terms "monoclonal antibody" or "monoclonal antibody composition" used herein and The term refers to a preparation of antibody molecules with a single molecular composition. Monoclonal antibody composition This demonstrates a single binding specificity and affinity for a particular epitope.

[0111] In this specification, the term "nucleic acid" is used interchangeably with the term "polynucleotide." Used to form deoxyribonucleotides or ribonucleotides and their single-stranded forms. This refers to polymers in a double-stranded form. The term "nucleic acid" refers to known nucleotides. Nucleic acids containing analogues or modified skeletal residues or modified links, which are synthetic nucleic acids, spontaneously occurring These are raw nucleic acids and non-naturally occurring nucleic acids, which have similar binding properties to the reference nucleic acid, and the reference nucleic acid It includes nucleic acids that are metabolized in a similar manner to rheotides. Examples of such analogues are, without limitation, To elaborate, phosphorothioates, phosphoramidates, methylphosphonates, chiral methyl It contains phosphonates, 2-O-methylribonucleotides, and peptide nucleic acids (PNA).

[0112] Unless otherwise indicated, a particular nucleic acid sequence is also a conservatively modified version of that sequence. It implicitly includes variants (e.g., degenerate codon substitutions) and complementary sequences, as well as explicitly including them. It also includes the indicated sequence. In particular, as detailed below, degenerate codon substitution is 1 The third position of one or more selected (or all) codons is a mixed base and / or Alternatively, this can be achieved by creating sequences substituted with deoxyinosine residues. (Batzer et al., Nucleic Acid Res. 19:5081, 1991; Ohtsuka et al., J. Biol. Chem. 260:2605 ~2608, 1985; and Rossolini et al., Mol. Cell. Probes 8:91~98, 1994).

[0113] The term "operably linked" refers to two or more polynucleotides (e.g., DN A) Refers to the functional relationship between segments. The term “operatably linked” is a transcription. Typically, this refers to the functional relationship between a regulatory sequence and the sequence being transcribed. For example, p The romor sequence or enhancer sequence is expressed in an appropriate host cell or other expression system. When stimulating or modulating the transcription of the code sequence within, it is operably linked to the code sequence. Generally, a promoter transcriptional regulatory sequence is operably linked to the sequence to be transcribed. It is physically adjacent to the sequence being transcribed, i.e., it is cis-acting. However, enhance Some transcriptional regulatory sequences, such as sensors, physically transfer their transcription to the coding sequences that enhance them. They don't need to be adjacent or placed in close proximity.

[0114] As used herein, the term "optimized" refers to a nucleotide sequence that has been optimized for production cells. Or producing organisms, generally eukaryotic cells, for example, Pichia cells, Chinese herbs In Muster ovary cells (CHO) or human cells, amino acids are produced using preferred codons. This means that it has been modified to encode an acid sequence. Optimized nucleotide sequence This is the amino that is originally encoded by the starting nucleotide sequence, which is also known as the "parent" sequence. The acid sequence is manipulated to be preserved completely or as much as possible. Optimization as described herein The sequence has been manipulated to have preferred codons in mammalian cells. Furthermore, this specification also describes the optimization of these sequences in other eukaryotic or prokaryotic cells. Expression is also planned. The amino acid sequence encoded by the optimized nucleotide sequence is also They also claim it is optimized.

[0115] In this specification, the terms “polypeptide” and “protein” refer to amino acid residues. The terms "polymer" and "protein" are used interchangeably. The term refers to the artificial chemical composition of one or more amino acid residues that correspond to naturally occurring amino acids. In addition to amino acid polymers that are natural mimics, there are also naturally occurring amino acid polymers and non-naturally occurring ones. This also applies to amino acid polymers. Unless otherwise indicated, certain polymers The ptydo sequence also implicitly includes its conservatively modified variants.

[0116] As used herein, the term “recombinant human antibody” refers to a human immunoglobulin gene. Regarding animals that are transgenic or transchromosomal (e.g., mice) Alternatively, the antibodies isolated from the hybridoma prepared in this way, or human antibodies, are expressed. A host cell transformed in such a way, for example, an antibody isolated from a transfectoma, Antibodies isolated from alternative combinatorial human antibody libraries, and human immunoglobulins. Splicing of all or part of the brin gene sequence into other DNA sequences Antibodies prepared, expressed, created, or isolated by any means or prepared, expressed, created, or isolated by recombinant means, Includes all human antibodies. Such recombinant human antibodies include framework regions and CDRs. The region has a variable region derived from the immunoglobulin sequence of human germline cells. However, In certain embodiments, such recombinant human antibodies induce in vitro mutagenesis. (Alternatively, if you use an animal that is transgenic for the human Ig sequence, in It can be subjected to vivo somatic mutagenesis, and therefore recombinant antibody VH The amino acid sequences of the region and VL region are derived from the VH and VL sequences of the human germline. These are related to the germline repertoire of in vivo human antibodies. This is a sequence that may not exist in nature.

[0117] The term "recombinant host cell" (or simply "host cell") refers to recombinant expression vectors. This refers to cells into which a ter has been introduced. Such terminology is intended to refer only to specific target cells. It is important to understand that this does not refer to the cells themselves, but rather to the descendant cells of such cells. In subsequent generations, certain modifications may occur due to mutations or environmental influences. Therefore, these offspring cells may not actually be identical to the parent cells. This falls within the scope of the term "host cell" as used in this specification.

[0118] The term "subject" includes humans and non-human animals. Non-human animals are non-human primates. (For example, crab-eating macaques), sheep, rabbits, dogs, cows, chickens, amphibians, and reptiles. This includes all vertebrates, such as reptiles (e.g., mammals and non-mammals). Except where otherwise specified, the terms “patient” and “subject” are used interchangeably in this specification. The terms "cynomolgus" or "cyno-molgus" used in the specification and The term refers to the crab-eating macaque (Cynomolgus monkey) (Macaca fascicularis). This refers to a person. In specific cases, the patient or subject is a human being.

[0119] In one embodiment, any disease or disorder (e.g., thromboembolic) used herein is described herein. Disorder) "To treat ~" or these terms "treatment" refer to improving a disease or disorder. To improve (i.e., to slow the onset of at least one of the disease or its clinical symptoms) It refers to (to do, stop, or mitigate). In another embodiment, it means "to treat ~ "Doing" or "treatment" includes parameters that may not be identifiable by the patient. This refers to mitigating or improving at least one physical parameter. In this context, "to treat ~" or "treatment" means to physically modulate a disease or disorder. It either rates (for example, stabilization of identifiable symptoms) or modulates physiologically (for example) (For example, stabilizing physical parameters) or modulating them physically and physiologically. In yet another embodiment, "~to treat" or "treatment" means a disease or This refers to preventing or delaying the occurrence, onset, or progression of a disorder.

[0120] For example, "prevention" in the context of the indications described herein, including thromboembolic disorders, means For example, in patients at risk of the aforementioned worsening, the thromboembolic disease described below This refers to any effect that prevents or slows the worsening of lameth.

[0121] The term "vector" refers to a vector that transports another polynucleotide to which it is linked. This is intended to refer to possible polynucleotide molecules. One type of vector is... "P" refers to a circular double-stranded DNA loop that can ligate various DNA segments. This is "Rasmid." Another type of vector uses a further DNA segment, the viral geno. Adeno-associated virus vectors (AAV or AAV2) that can ligate to the virus Which viral vector is it? A certain type of vector (for example, a bacterial origin of replication). Vectors (and mammalian episomal vectors) are introduced into the host cells into which they are... It is capable of self-replication. Other vectors (e.g., non-mammalian episomal vectors) When introduced into a host cell, it can be incorporated into the host cell's genome, thereby allowing the host They replicate together with the main genome. Furthermore, certain types of vectors are genes that are operationally linked to each other. It is possible to direct the expression of genes. In this specification, such vectors are referred to as "combinations." This is called a recombinant expression vector (or simply, an expression vector). Generally, recombinant DN In method A, useful expression vectors are often in the form of plasmids. Plasmids Since it is the most commonly used form of vector, in this specification we will refer to it as "plasmid" and " It can be used interchangeably with "Vector". However, the present invention provides equivalent functionality. Viral vectors (e.g., replication-deficient retroviruses, adenoviruses, and adenoviruses) This is intended to include other forms of expression vectors, such as associated viruses. [Brief explanation of the drawing]

[0122] [Figure 1-1] Figures 1A-1C show the effect of NOV1401 on FeCl3-induced thrombosis in FXI- / - mice reconstituted with human FXI protein. NOV1401 inhibited thrombosis in a dose-dependent manner. The antibody prolonged aPTT to the same extent as in untreated FXI- / - mice. [Figure 1-2] (As stated above.) [Figure 2] Figures 2A-2B show the relationship between the effect of multiple intravenous (iv) (A; N=2) or subcutaneous (sc) (B; N=2) administrations of NOV1401 at doses of 3 mg / kg, 10 mg / kg, and 30 mg / kg on aPTT (diamond) and total plasma NOV1401 levels (square) in cynomolgus monkeys. A single dose of 3 mg / kg resulted in approximately a twofold increase in aPTT, which was maintained for 5-6 weeks. All test doses prolonged aPTT to a similar degree, and higher test doses did not appear to increase the magnitude of the aPTT prolongation observed at the 3 mg / kg dose. [Figure 3]Figures 3A-3B show the relationship between the effects of multiple IV (A; N=2) or sc (B; N=2) administrations of NOV1401 at doses of 3 mg / kg, 10 mg / kg, and 30 mg / kg on plasma free FXI (square) and aPTT (diamond) in cynomolgus monkeys. A single dose of 3 mg / kg reduced free FXI by approximately 90% over 5-6 weeks. All test doses reduced free FXI to a similar degree, and higher test doses did not appear to increase the magnitude of the reduction in free FXI observed at the 3 mg / kg dose. [Figure 4] Figures 4A-4B show the X-ray structures of the Fab of the NOV1401 antibody of the present invention bound to FXI. Figure 4A shows the X-ray structure of the NOV1401 Fab-FXI CD complex. The FXI catalytic domain is shown as a gray surface, and the Fab is shown as light gray (light chain) and dark gray (heavy chain) ribbons. Figure 4B shows the X-ray structure of the NOV1401 Fab-FXI CD complex superimposed on Zymogen FXI. The FXI catalytic domain is shown as a gray ribbon. The variable domains of the Fab are shown as light gray (VL) and dark gray (VH) ribbons. The Zymogen structure containing four apple domains is superimposed as a dark gray ribbon at the bottom of the structure (PDB 2F83). The activation cleavage site (Ile370) is indicated. [Figure 5] Figures 5A and 5B show the structural changes of FXIa upon binding to NOV1401 Fab. Figure 5A shows a visual view of the active site of FXIa before Fab binding. FXIa is represented as a ribbon with a transparent surface. Cross-sections of the structures that change conformation upon Fab binding are labeled (loops 145, 188, and 220). S1 and the subpocket of S1 are pointed to. Figure 5B shows the inactive conformation of FXI within the Fab complex. (Fab is not shown). [Figure 6-1]Figures 6A-6C show composite response curves for anti-FXI / FXIa antibodies. Figure 6A shows inhibition of factor XIa activity by NOV1401. This is a representative composite response curve for NOV1401, an antibody that inhibits the enzyme activity of full-length human FXIa. The assay measures the cleavage of a fluorescently labeled peptide, as described in Example 3. A nonlinear curve fitting using the logistic fitting model [y=A2+(A1-A2) / (1+(x / IC50)p)[wherein y is the inhibition percentage when the inhibitor concentration is x, A1 is the minimum inhibition value, A2 is the maximum inhibition value, and the exponent p is the Hill coefficient] is obtained for this representative dataset, yielding an IC50 value of 160 pM. Figure 6B shows a composite response curve for aPTT. This is a representative composite response curve for prolongation of coagulation time by the antibody NOV1401 in an aPTT assay using pooled human plasma. The assay measures coagulation time after initiating the intrinsic coagulation cascade in the presence of different concentrations of NOV1401, as described in Example 4. The black line represents the fit using a logistic nonlinear fitting model. The dotted line represents the baseline coagulation time of pooled human plasma in the absence of NOV1401. The baseline coagulation time is 32.3 seconds, indicated by the gray dashed line in the graph. The gray dotted line indicates the antibody concentration at which the coagulation time doubled compared to baseline, i.e., reached a value of 2 × aPTT, which is 14 nM. Figure 6C shows the TGA response curve. It shows a representative composite response curve for inhibition of thrombin generation by the antibody NOV1401 in TGA with pooled human plasma. The assay, as described in Example 4, measures the effect of different concentrations of NOV1401 on thrombin generation that can be induced by very low concentrations of tissue factor (TF) via a so-called thrombin-to-FXIa feedforward loop. The black line represents the fit using a four-parameter dose-response curve model. The dotted line represents the residual thrombin concentration resulting from thrombin generation induced by small amounts of TF. The IC50 value of 24 nM and the residual thrombin concentration of 159 nM (dotted line) were calculated for this composite response curve. [Figure 6-2] (As stated above.) [Figure 7-1] Figures 7A and 7B show the effects of NOV1401 on aPTT and FXI activity (FXI:C) with sc administration at 10 mg / kg (N=3) and 100 mg / kg (N=5) weekly for 13 weeks (14 doses), or IV administration at 50 mg / kg (N=3) weekly for 4 weeks (5 doses). Figure 7A shows the effect on aPTT as measured on days 2, 23, and 79 of the study. aPTT increased 2.1 to 3 times in all animals treated with NOV1401 and remained high throughout the administration period of the study. No dose-dependency was observed, and no sex-related differences were found. Figure 7B shows the effect on FXI:C as measured on days 2, 23, and 79 of the study, depicted as a percentage of plasma FXI activity. FXI:C levels decreased to 5–12% in all animals treated with NOV1401 and remained at these levels throughout the study's administration period. No dose-dependent differences were observed, nor were there any sex-related differences. [Figure 7-2] (As stated above.)

[0123] Detailed description The present invention partially relates to an antibody that specifically binds to FXIa and inhibits its biological activity. Based on the discovery of this child, the present invention relates to full-length IgG format antibodies, as well as Fab fragments (e.g.) For example, regarding antibodies NOV1090 and NOV1401, both of their antigen-binding fragments... ru.

[0124] Therefore, the present invention relates to FXI and / or FXIa (e.g., humans, rabbits, and Antibodies that specifically bind to FXI and / or FXIa) of cynomolgus monkeys, pharmaceutical compositions This provides methods for producing and using such antibodies and compositions.

[0125] Factor XI FXI occurs in both the intrinsic and extrinsic coagulation pathways, as well as in plasma. It plays an important role in bridging the initial and amplification phases of hemostasis. Factor XIIa and Both thrombins, as a result of activating FXI, lead to sustained thrombin production. It can lead to inhibition of fibrinolysis. FXI is used in a high-tissue-factor environment "after vascular injury". Although it plays a small role in normal hemostasis, it is thought to play a key role in thrombosis. Severe factor XI deficiency is associated with a reduced incidence of ischemic stroke and venous thromboembolic events. Related to (Salomon et al 2008; Salomon, et al. (2011) Thromb Haemost.; 105:269- 73). Bleeding symptoms in subjects with severe factor XI deficiency are infrequent and mild. It is often damage-inducing, and increases fibrinolytic activity in the oral mucosa, nasal mucosa, and urinary tract. It is preferable that the tissue is affected (Salomon et al 2011). Bleeding in life-threatening organs is It is either extremely rare or not recognized.

[0126] Plasma coagulation is a sequential process in which coagulation factors in the blood interact and become activated, and the final In essence, this results in the production of fibrin and the formation of a blood clot. In the genotype model, the process of fibrin formation involves two distinctly different pathways, namely the intrinsic pathway. It can be initiated by both the pathway and the extrinsic pathway (Mackman, 2008).

[0127] In the extrinsic pathway, vascular injury leads to the production of extravascular tissue factor (TF) and factor VII (FVII). It interacts with and activates it, thereby activating factor X and prothrombin. It brings about a sequential transformation. Active thrombin ultimately converts soluble fibrinogen into fibrin. It then converts to n. The extrinsic pathway is central to hemostasis, and the influence of coagulation factors within this pathway Wataru carries the risk of bleeding as a result.

[0128] In the intrinsic pathway, factor XII is sometimes activated through a process called contact activation. It is possible. The production of activated factor XIIa is due to the sequential activation of factor XI and factor IX. It brings about a change. Factor IXa activates Factor X, thus affecting both the extrinsic and intrinsic pathways. The pathways merge at this stage (common pathway). Thrombin activity is when thrombin is factor XI. The child is activated independently of factor XII via a feedforward loop. This is boosted by amplifying its own generation. This feedforward loop is It contributes to sustained thrombus growth, but coagulation requires strong activation by extravascular tissue factor. Since this is sufficient, its involvement in hemostasis is limited to a minimum. Therefore, the intrinsic pathway is... It is not substantially involved in blood (Gailani and Renne (2007) Arterioscler Thromb Vasc Biol 2007, 27(12):2507-13, Muller, Gailiani, and Renne 2011).

[0129] Preclinical studies using various methods to inhibit FXI or FXIa across various species However, this has contributed to the validation of this target. FXI- / - mice were experimentally shown to have venous thrombosis (Wa ng, et al. (2006) J Thromb Haemost; 4:1982-8) and arterial thrombosis (Wang, et al. (20 05) It is resistant to J Thromb Haemost; 3:695-702). Treatment of mice with an antibody that blocks activation (Ab; 14E11) resulted in experimentally observed thrombosis. This results in inhibition (Cheng, et al. (2010) Blood, 116:3981-9), and ischemic stroke. Reduced stroke size in a mouse model of [the substance] (Leung, et al. (2012) Transl Stroke Res 2012; 3:381-9). Anti-FXI blocking FXIa binding and activation. In baboons administered Ab, on collagen-coated vascular grafts, A reduction in the proliferation of platelet-rich thrombi was observed (Tucker, et al. (2009) Blood 2009; 113:93). 6-44) Similar results were also found in this model by 14E11 (Cheng (2010). No excessive bleeding was observed in any of these studies.

[0130] Mice (Zhang, et al. (2010) Blood 2010; 116:4684-92), cynomolgus monkeys (Younis, et al. (2012) Blood 2012; 119:2401-8), and baboons (Crosby, et al. (2013) Arteri In oscler Thromb Vasc Biol 2013; 33:1670-8), antisense oligonucleotides Blocking the synthesis of FXI by this method provides antithrombotic and anti-inflammatory effects without excessive bleeding. This resulted in a coagulation effect. Furthermore, in rats (Schumacher, et al. (2007) Eur J Pharmacol 2007; 570:167-74) and rabbits (Wong, et al. (2011) J Thromb Thromboly In the venous thrombosis model and arterial thrombosis model described in sis 2011; 32:129-37), low molecular weight Blocking FXIa with inhibitors has yielded similar results.

[0131] Patients with severe FXI deficiency rarely experience spontaneous bleeding and have high fibrinolytic activity. Except for severe tissue bleeding, it is limited to mild trauma-induced bleeding. Severe FXI deficiency is rare. The study aims to clarify the thrombotic profiles of these patients compared to the general population. This makes the use of research inevitable. In particular, such research shows that in these patients, ischemic stroke (Salomon 2008) and deep vein thrombosis (DVT) (Salomon, et al. (2011) Blood 200 8; 111: 4113-17) The incidence of severe FXI is reported to be reduced. The number of ischemic strokes observed in 115 patients with deficiency (N=1) is, The number of cases was lower than the predicted number in El's general population (N=8.6) (p<0.003). The number of DVT cases in patients with severe FXI deficiency (N=0) was lower than in the control population. The number of cases was lower than the predicted number (N=4.7) (p<0.019). Conversely, FXI Individuals whose Bell number is above the 90th percentile had twice the risk of developing DVT (Meijer). (2000) N Engl J Med. 2000; 342:696-701).

[0132] In recent years, patients undergoing total knee arthroplasty, a procedure that predisposes them to DVT, have been referred to as FXI AN. Antisense therapy or standard treatment (enoxaparin) was used. Antisense group (300m g) Compared to standard treatment, the incidence of venous thrombosis was reduced by one-seventh, and bleeding events decreased. (Not statistically significant) was shown (Buller et al, (2014) N Engl J Med. 372(3):232-40. i: 10.1056 / NEJMoa1405760. Epub 2014 Dec 7).

[0133] In summary, the above studies strongly support FXI as a valid target for antithrombotic therapy.

[0134] FXIa antibody and antigen-binding fragments This invention provides antibodies that specifically bind to FXI and / or FXIa. In some embodiments, the present invention relates to the FXI and / or other forms of humans, rabbits, and cynomolgus macaques. The present invention provides an antibody that specifically binds to FXIa as well. This includes, but is not limited to, human monoclonal antibodies and Fab isolated as described above. stomach.

[0135] The present invention relates to FXI protein and / or FXIa protein (e.g., human, u Antibodies that specifically bind to FXI and / or FXIa) in herons and cynomolgus monkeys. The present invention provides an antibody containing a VH domain having the amino acid sequences of SEQ ID NOs. 9 and 29. The present invention also specifically binds to FXI protein and / or FXIa protein. The antibody that matches one of the VH CDRs listed in Table 1 below. The present invention also provides antibodies containing VH CDRs having an acid sequence. In particular, the present invention provides antibodies containing FXI protein F(x) and / or F(x)a proteins (e.g., F(x)a proteins of humans, rabbits, and cynomolgus monkeys) Antibodies that specifically bind to XI and / or FXIa), as listed in Table 1 below. One, two, or three or more VHs having any of the amino acid sequences of VH CDRs The present invention provides antibodies containing (or, alternatively, composed of) CDRs.

[0136] The present invention is an antibody that specifically binds to the FXIa protein, and is related to SEQ ID NO: 19 or The present invention provides an antibody containing a VL domain having a 39-amino acid sequence. Proteins and / or FXIa proteins (e.g., human, rabbit, and crab meat) Antibodies that specifically bind to FXI and / or FXIa) of sieves, as shown in Table 1 below. Includes a VL CDR having any one of the amino acid sequences listed among the VL CDRs. The present invention also provides antibodies. In particular, the present invention provides FXIa proteins (for example, human, rabbit, and An antibody that specifically binds to FXI and / or FXIa) of cynomolgus monkeys, One or two VL CDRs having one of the amino acid sequences listed in Table 1 below. The present invention provides an antibody containing (or, alternatively, consisting of) three or more VL CDRs. ru.

[0137] Other antibodies of the present invention have been mutated, but within the CDR region, as listed in Table 1... The CDR region depicted within the sequence, and at least 60, 70, 80, 85, 90, or contains amino acids having 95 percent identity. In some embodiments, the present invention Other antibodies, within the CDR region, depict the CDR region within the sequence listed in Table 1 and In comparison, one, two, three, four, or five or fewer amino acids have been mutated. Includes the amino acid sequence of the variant.

[0138] The present invention also relates to FXI protein and / or FXIa protein (e.g., human, VH, VL, and full-length weight of antibodies that specifically bind to rabbit and cynomolgus monkey FXIa). We also provide nucleic acid sequences that encode the chain and the full-length light chain. Such nucleic acid sequences are mammalian. The expression in cells can be optimized (for example, Table 1 shows the weight of the antibody of the present invention). (Shows nucleic acid sequences optimized for the chain and light chain.)

[0139] [Table 1-1]

[0140] [Table 1-2]

[0141] [Table 1-3]

[0142] [Table 1-4]

[0143] [Table 1-5]

[0144] [Table 1-6]

[0145] Other antibodies of the present invention involve mutating amino acids or nucleic acids encoding amino acids. However, the sequences listed in Table 1 still contain at least 60, 65, 70, 75, 80, 85, It contains antibodies having 90 or 95 percent identity. Some embodiments are variable Within the region, while maintaining substantially the same antigen-binding activity, the sequences described in Table 1 are drawn. When compared to the indicated variable region, there are one, two, three, four, or five or fewer amino acids. It contains mutant amino acid sequences in which the acid has been mutated.

[0146] Each of these antibodies can bind to FXI and / or FXIa, so the VH sequence , VL sequence, full-length light chain sequence, and full-length heavy chain sequence (amino acid sequence and amino acid sequence are coded The nucleotide sequence to be used is "mixed and matched" with the other FXI-binding antibody of the present invention. It is possible to create antibodies that bind to the body and / or FXIa. Such "mixed, Matched FXI-binding antibodies and / or FXIa-binding antibodies are used in this technology. Known binding assays (e.g., ELISA and other assays described in the Examples section) It can be used to investigate. When mixing and matching these strands, a specific VH / V The VH sequence derived from L pairing will be replaced with a structurally similar VH sequence. In addition, a full-length heavy chain sequence derived from a specific full-length heavy chain / full-length light chain pair is structurally similar to a full-length heavy chain. The chain sequence will be replaced. Similarly, the VL sequence derived from a specific VH / VL pairing will be replaced. The VL sequence will be replaced with a structurally similar one. Similarly, specific full-length heavy chains / full-length light chains will be replaced. The full-length light chain sequence derived from pairing will be replaced with a structurally similar full-length light chain sequence. .

[0147] Therefore, in one embodiment, the present invention is selected from the group consisting of Sequence IDs 9 and 29. From the group consisting of heavy chain variable domains containing amino acid sequences, and SEQ ID NOs: 19 and 39 Isolated antibody having a light chain variable domain containing a selected amino acid sequence or its antigen-binding properties Provides a region, in this case the antibody is FXI and / or FXIa (e.g., human, u It specifically binds to herons and cynomolgus macaques (FXIa).

[0148] More specifically, in certain embodiments, the present invention relates to Sequence IDs 9 and 29; or 19 and Heavy chain variable domains and light chain variable domains containing amino acid sequences selected from each of the 39. This provides an isolated antibody or its antigen-binding region having a main component.

[0149] In a specific embodiment, this specifically binds to human FXI and / or FXIa. The antibody or antigen-binding fragment presented in the specification comprises the amino acid sequence of SEQ ID NO: 9. It includes a heavy chain variable region and a light chain variable region containing the amino acid sequence of SEQ ID NO: 19.

[0150] In a specific embodiment, this specifically binds to human FXI and / or FXIa. The antibody or its antigen-binding fragment presented in the specification contains the amino acid sequence of SEQ ID NO: 29 It includes a heavy chain variable region and a light chain variable region containing the amino acid sequence of SEQ ID NO: 39.

[0151] In another aspect, the present invention relates to (i) an ami optimized for expression in mammalian cells. An amino acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11 or 31. This is the full-length heavy chain, as well as an amino acid sequence optimized for expression in mammalian cells. This refers to a full-length light chain containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 21 or 41. (ii) to provide an isolated antibody having, or a functional protein containing the antigen-binding moiety thereof. More specifically, in certain embodiments, the present invention relates to Sequence IDs 11 and 31; or 19. Isolated antibodies having heavy and light chains containing amino acid sequences selected from each of 39. or provides the antigen-binding region thereof.

[0152] In a specific embodiment, this specifically binds to human FXI and / or FXIa. The antibody or antigen-binding fragment presented in the specification contains the amino acid sequence of SEQ ID NO: 11 It contains a heavy chain and a light chain containing the amino acid sequence of SEQ ID NO: 21.

[0153] In a specific embodiment, this specifically binds to human FXI and / or FXIa. The antibody or antigen-binding fragment presented in the specification contains the amino acid sequence of SEQ ID NO: 31 It includes a heavy chain variable region and a light chain variable region containing the amino acid sequence of SEQ ID NO: 41.

[0154] As used herein, the terms “complementarity-determining region” and “CDR” refer to antigen specificity. This refers to the sequence of amino acids within the antibody variable region that confer antigen-binding affinity. Generally, within each heavy chain variable region (HCDR1, HCDR2, HCDR3), there are three CDRs. There are three CDRs within each light chain variable region (LCDR1, LCDR2, LCDR3). be.

[0155] The precise amino acid sequence boundaries of a given CDR are defined in Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National In Institutes of Health, Bethesda, MD ("Kabat" numbering scheme), Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme), Lefranc e According to t al., (2003) Dev. Comp. Immunol., 27, 55-77 ("IMGT" numbering scheme) The schemes described, or the "combination" systems, and many other well-known schemes. You can easily make a decision using one of these options.

[0156] For example, according to Kabat, NOV109 is an antibody located within the heavy chain variable domain (VH). The CDR amino acid residues of 0 are 31-35 (HCDR1), 50-66 (HCDR2), They are numbered 99-111 (HCDR3) and are located within the light chain variable domain (VL). The CDR amino acid residues are 22-35 (LCDR1), 51-57 (LCDR2), and They are numbered 90-100 (LCDR3). According to Chothia, within VH CDR amino acids are 26-32 (HCDR1), 52-57 (HCDR2), and 99 It is numbered ~111 (HCDR3), and the amino acid residues in VL are 25~33 ( Numbered as LCDR1), 51-53 (LCDR2), and 92-99 (LCDR3). This involves combining CDR definitions from both Kabat and Chothia. Furthermore, CDRs are amino acid residues 26-35 (HCDR1) and 50-66 (H) in human VH. CDR2), and 99-111 (HCDR3), as well as two amino acid residues in human VL. 2-35 (LCDR1), 51-57 (LCDR2), and 90-100 (LCDR3) ) consists of combining CDR definitions from both Kabat and Chothia. Furthermore, the "combination" CDR is the amino acid residues 26-35 (HCDR1) and 50 in human VH. ~66 (HCDR2), and 99~108 (HCDR3), as well as mycelium in human VL. 24-38 (LCDR1), 54-60 (LCDR2), and 93-101 ( It consists of LCDR3). Another example, according to IMGT, is within the heavy chain variable domain (VH). The CDR amino acid residues are 26-33 (HCDR1), 51-58 (HCDR2), and They are numbered 97-108 (HCDR3), and are C within the light chain variable domain (VL). The DR amino acid residues are 27-36 (LCDR1), 54-56 (LCDR2), and 9 They are numbered 3-101 (LCDR3). Table 1 shows examples of anti-FXI / FXIa antibodies. For example, regarding NOV1090 and NOV1401, Kabat, Chothia, and Examples of HCDR1, HCDR2, HCDR3, and LCDR by IMGT. 1, LCDR2, and LCDR3 are presented. In another embodiment, the present invention is described in Table 1. The heavy and light chains CDR1, CDR2, and CDR3, or combinations thereof. This provides an FXIa-binding antibody containing [a specific component]. The amino acid sequence of the antibody's VH CDR1 is [a specific component]. Shown in numbers 3 and 23. The amino acid sequences of the antibody's VH CDR2 are shown in SEQ ID NOs: 4 and 2 As shown in 4. The amino acid sequence of the antibody VH CDR3 is shown in SEQ ID NOs. 5 and 25. The amino acid sequences of the antibody VL CDR1 are shown in SEQ ID NOs: 13 and 33. The amino acid sequences of the antibody's VL CDR2 are shown in SEQ ID NOs: 14 and 34. The amino acid sequences of the antibody's VL CDR3 are shown in SEQ ID NOs. 15 and 35. These CDR areas are represented using the Kabat system.

[0157] Alternatively, the Chothia system (Al-Lazikani et al., (1997), JMB 273 927~948) As specified using, the amino acid sequence of the antibody VH CDR1 is as follows: SEQ ID NO: 6 and The amino acid sequence of the antibody VH CDR2 is shown in 26. The amino acid sequences of the antibody's VH CDR3 are shown in SEQ ID NOs. 8 and 28. The amino acid sequences of the antibody VL CDR1 are shown in SEQ ID NOs. 16 and 36. The amino acid sequences of the antibody's VL CDR2 are shown in SEQ ID NOs: 17 and 37. The amino acid sequences of the antibody's VL CDR3 are shown in SEQ ID NOs. 18 and 38.

[0158] Alternatively, the amino acid of the antibody VH CDR1 as defined using a combination system. The acid sequence is shown in SEQ ID NO: 46. The amino acid sequence of the antibody VH CDR2 is: It is shown in number 4. The amino acid sequence of the antibody VH CDR3 is shown in SEQ ID NO: 5. The amino acid sequence of the antibody VL CDR1 is shown in SEQ ID NO: 33. The amino acid sequence of L CDR2 is shown in SEQ ID NO: 14. The amino acid sequence is shown in SEQ ID NO: 15.

[0159] Alternatively, the VH CDR of the antibody as defined using the IMGT numbering scheme. The amino acid sequence of 1 is shown in SEQ ID NO: 43. The amino acid sequence of the antibody VH CDR2 The sequence is shown in SEQ ID NO: 44. The amino acid sequence of the antibody VH CDR3 is SEQ ID NO: It is shown in 45. The amino acid sequence of the antibody VL CDR1 is shown in SEQ ID NO: 47. The amino acid sequence of the antibody VL CDR2 is shown in SEQ ID NO: 37. The amino acid sequence of L CDR3 is shown in SEQ ID NO: 15.

[0160] Each of these antibodies is capable of binding to FXI and / or FXIa. Antigen binding specificity is mainly determined by the CDR1, CDR2, and CDR3 regions. Given that, each antibody is a component of the other FXI-binding molecule of the present invention and / or To create FXIa-binding molecules, VH CDR1, VH CDR2, and VH C This contains DR3, as well as VL CDR1, VL CDR2, and VL CDR3. This is preferable, but VH CDR1 sequence, VH CDR2 sequence, and VH CDR3 sequence , as well as the VL CDR1 sequence, VL CDR2 sequence, and VL CDR3 sequence, It is possible to "mix and match" (i.e., mix CDRs derived from different antibodies). (and can be matched). Such "mixed and matched" FXI binding components The child and / or FXIa-binding antibody can be used in binding assays and procedures known in the art. The binding assays described in the example (e.g., ELISA, SET, BIACORE®) This can be investigated using (Say). When mixing and matching VH CDR sequences. This is a CDR1 sequence, CDR2 sequence, and / or CDR3 sequence derived from a specific VH sequence. The columns will be replaced with structurally similar CDR sequences. Similarly, the VL CDR sequences When mixing and matching, the CDR1 and CDR2 sequences derived from a specific VL sequence , and / or the CDR3 sequence shall be replaced with a structurally similar CDR sequence. Those skilled in the art can use novel VH and VL sequences to represent one or more VH CDR regions. The column and / or VL CDR region sequence for the monoclonal antibody of the present invention Created by substituting a structurally similar sequence derived from the CDR sequence shown in the detailed document. It will be readily apparent that this is possible. In addition to the foregoing, in one embodiment, as described herein The antigen-binding fragments of the antibodies are VH CDR1, VH CDR2, and VH CDR 3, or possibly including VL CDR1, VL CDR2, and VL CDR3 Yes, in this case the fragment is a single variable domain in FXI and / or FXIa. They combine.

[0161] In certain embodiments of the present invention, antibodies or antigen-binding fragments thereof are as shown in Table 1. It may have the heavy and light chain sequences of the Fab. More specifically, an antibody or its antigen. The binding fragment may have the heavy and light chain sequences of NOV1090 and NOV1401.

[0162] In other embodiments of the present invention, an antibody specifically binding to FXI and / or FXIa is used. Alternatively, the antigen-binding fragment is defined by Kabat and listed in Table 1, and is heavy-chain variable. CDR1 region, CDR2 heavy chain variable region, CDR3 heavy chain variable region, C light chain variable region The present invention further includes DR1, CDR2 of the light chain variable region, and CDR3 of the light chain variable region. In other embodiments, an antibody or antigen that specifically binds to FXI and / or FXIa. The binding fragments are defined by Chothia and are described in Table 1 as heavy chain variable regions. CDR1 in the region, CDR2 in the heavy chain variable region, CDR3 in the heavy chain variable region, CD in the light chain variable region Includes R1, CDR2 of the light chain variable region, and CDR3 of the light chain variable region. The antibody or antigen-binding fragment that specifically binds to FXI and / or FXIa is a combination of the two. The CDR1 and heavy chain variable regions are defined by the matching system and are listed in Table 1. CDR2 of the chain variable region, CDR3 of the heavy chain variable region, CDR1 of the light chain variable region, light chain variable region This includes the CDR2 region and the CDR3 region of the light chain variable region. In yet another embodiment of the present invention, Antibodies or antigen-binding fragments that specifically bind to FXI and / or FXIa are IM The heavy chain variable region CDR1, defined by GT and described in Table 1, and the heavy chain variable region CDR2 in the region, CDR3 in the heavy chain variable region, CDR1 in the light chain variable region, CD in the light chain variable region Includes R2 and the light chain variable region CDR3.

[0163] In specific embodiments, the present invention specifically binds to FXI and / or FXIa. The antibody is characterized by CDR1 in the heavy chain variable region of SEQ ID NO: 3 and CD in the heavy chain variable region of SEQ ID NO: 4. R2; CDR3 of the heavy chain variable region of SEQ ID NO: 5; CDR1 of the light chain variable region of SEQ ID NO: 13; CDR2 of the light chain variable region of SEQ ID NO: 14; and CDR3 of the light chain variable region of SEQ ID NO: 15 Contains antibodies that include [specific component].

[0164] In specific embodiments, the present invention specifically binds to FXI and / or FXIa. The antibody is such that the heavy chain variable region CDR1 of SEQ ID NO: 23 and the heavy chain variable region of SEQ ID NO: 24 CDR2 of the heavy chain variable region of sequence number 25, CDR3 of the light chain variable region of sequence number 33 DR1, CDR2 of the light chain variable region of sequence number 34, and the light chain variable region of sequence number 35 Contains antibodies including CDR3.

[0165] In specific embodiments, the present invention specifically binds to FXI and / or FXIa. The antibody is such that CDR1 is in the heavy chain variable region of SEQ ID NO: 6, and C is in the heavy chain variable region of SEQ ID NO: 7. DR2, CDR3 of the heavy chain variable region of SEQ ID NO: 8, and CDR1 of the light chain variable region of SEQ ID NO: 16 , CDR2 of the light chain variable region of SEQ ID NO: 17, and CDR of the light chain variable region of SEQ ID NO: 18 Contains antibodies that include 3.

[0166] In specific embodiments, the present invention specifically binds to FXI and / or FXIa. The antibody is such that the heavy chain variable region CDR1 of SEQ ID NO: 26 and the heavy chain variable region of SEQ ID NO: 27 CDR2 of the heavy chain variable region of sequence number 28, CDR3 of the light chain variable region of sequence number 36 DR1, CDR2 of the light chain variable region of sequence number 37, and the light chain variable region of sequence number 38 Contains antibodies including CDR3.

[0167] In specific embodiments of this specification, binding specifically occurs to FXI and / or FXIa. An antibody, wherein the heavy chain variable region CDR1 of SEQ ID NO: 43; the heavy chain variable region of SEQ ID NO: 44 CDR2; Heavy chain variable region of SEQ ID NO: 45; CDR3; Light chain variable region of SEQ ID NO: 47 R1; CDR2 of the light chain variable region of SEQ ID NO: 37; and C of the light chain variable region of SEQ ID NO: 15 Antibodies containing DR3 are presented.

[0168] In specific embodiments of this specification, binding specifically occurs to FXI and / or FXIa. The antibody is characterized by CDR1 in the heavy chain variable region of SEQ ID NO: 46; and C in the heavy chain variable region of SEQ ID NO: 4. DR2; CDR3 of the heavy chain variable region of SEQ ID NO: 5; CDR1 of the light chain variable region of SEQ ID NO: 33 ;CDR2 of the light chain variable region of SEQ ID NO: 14; and CDR of the light chain variable region of SEQ ID NO: 15 Antibodies containing 3 are presented.

[0169] In certain embodiments, the present invention relates to the FXI and / or FX described in Table 1. It contains an antibody or antigen-binding fragment that specifically binds to Ia. In preferred embodiments, FX The antibody or antigen-binding fragment that binds to I and / or FXIa is NOV1090 and It is NOV1401.

[0170] The variable region or full-length chain of the antibody uses human germline immunoglobulin genes. When obtained from a system, the human antibodies used herein are produced from a specific germline sequence. If it is "a thing" or "derived from" it, heavy chain or light chain variable region or full-length heavy chain It contains a light chain. Such a system is a transgenic organism that possesses human immunoglobulin genes. The mice are immunized with the target antigen, or the antigen is presented on a phage containing the target antigen. This includes screening a library of human immunoglobulin genes. Human antibodies that are "products" of or "derived from" a cell lineage immunoglobulin sequence are The amino acid sequence of human antibodies was compared with the amino acid sequence of human germline immunoglobulins. , human germline antibody whose sequence most closely resembles the sequence of a human antibody (i.e., has the highest identity %) By selecting the immunoglobulin sequence of the cell lineage, it can be identified as such. ru.

[0171] It is a "product" of or derived from an immunoglobulin sequence of a specific human germ cell lineage. Human antibodies that "do" are, for example, intentionally produced by spontaneous somatic mutations or site-directed mutations. Due to the introduction of this gene, it may contain differences in amino acids compared to the germline sequence. In the VH framework region or VL framework region, the selected human antibody is The amino acid sequence is encoded by the immunoglobulin genes of the human germline. The sequence is at least 90% identical to that of human antibodies, and the germline immunoglobulins of other species When compared to an amino acid sequence (e.g., a mouse germline sequence), it can be identified as human. Typically, they contain amino acid residues. In some cases, human antibodies contain amino acids. The amino acid sequence is the amino acid sequence encoded by the germline immunoglobulin gene, At least 60%, 70%, 80%, 90%, or at least 95%, or less At the very least, they can be 96%, 97%, 98%, or even 99% identical.

[0172] Recombinant human antibodies are human antibodies within the VH framework region or VL framework region. 10 amino acids with the amino acid sequence encoded by germline immunoglobulin genes Typically, it presents differences below the acid level. In certain cases, human antibodies are germ cells. The amino acid sequence encoded by the immunoglobulin gene series is 5 amino acids or less. It may present differences of 4, 3, 2, or 1 amino acid or less. Human germline Examples of immunoglobulin genes include the germline variable domain fragments described below, as well as This includes, but is not limited to, DP47 and DPK9.

[0173] homologous antibody In yet another embodiment, the present invention relates to the sequences listed in Table 1 (for example, sequence number 2). An antibody or its antigen-binding properties containing an amino acid sequence homologous to 9, 31, 39, or 41) The antibody provides fragments of the FXI protein and / or FXIa protein (for example) It binds to human FXIa, rabbit FXIa, and cynomolgus monkey FXIa, and NOV1 The antibodies listed in Table 1, such as 090 and NOV1401, retain their desired functional properties. In a specific example, such homologous antibodies are CDR amino acids as listed in Table 1. Acid sequences (for example, Kabat CDR, Chothia CDR, IMGT CDR, It holds the combination CDR.

[0174] For example, the present invention relates to an isolated antibody or so comprising a heavy chain variable domain and a light chain variable domain. It provides a functional antigen-binding fragment, in which case the heavy chain variable domain is SEQ ID NO: 9 and An amino acid sequence selected from a group of 29, and at least 80%, at least 90%, or containing at least 95% identical amino acid sequences; the light chain variable domain is SEQ ID NO: 19 and an amino acid sequence selected from the group consisting of 39, and at least 80%, at least 9 Containing 0% or at least 95% identical amino acid sequences; the antibody is FXI and / or This applies to FXIa (e.g., human FXIa, rabbit FXIa, and cynomolgus macaque FXIa). It binds specifically. In one embodiment, the isolated antibody or its functional antigen-binding fragment is heavy It includes a chain variable domain and a light chain variable domain, in which case the heavy chain variable domain is sequence number The amino acid sequence of No. 9, and at least 80%, at least 90%, or at least 95% It contains the same amino acid sequence; the light chain variable domain has the same amino acid sequence as SEQ ID NO: 19 and a small amount Containing at least 80%, at least 90%, or at least 95% identical amino acid sequences; Antibodies include FXI and / or FXIa (e.g., human FXIa, rabbit FXIa, and It specifically binds to cynomolgus monkeys (FXIa). In one embodiment, the isolated antibody or its mechanism The active antigen-binding fragment includes a heavy chain variable domain and a light chain variable domain, in this case, The heavy chain variable domain has the amino acid sequence of SEQ ID NO: 29 and at least 80% and at least 9 Containing 0% or at least 95% identical amino acid sequences; the light chain variable domain is sequence number The amino acid sequence of No. 39 and at least 80%, at least 90%, or at least 95% % Contains identical amino acid sequences; the antibody is FXI and / or FXIa (e.g., human F It specifically binds to XIa, rabbit FXIa, and cynomolgus monkey FXIa. In certain embodiments, the heavy and light chain sequences are defined by Kabat, HCDR1 sequence Column, HCDR2 sequence, HCDR3 sequence, LCDR1 sequence, LCDR2 sequence, and LCD The R3 sequence further includes, for example, sequence numbers 3, 4, 5, 13, 14, and 15, respectively. In certain other aspects of the present invention, the heavy chain sequence and the light chain sequence are provided by Chothia. The specified sequences are HCDR1, HCDR2, HCDR3, LCDR1, and LC. DR2 sequences and LCDR3 sequences, for example, sequence numbers 6, 7, 8, 16, and 1, respectively. The following further include 7 and 18. In certain other embodiments, the heavy chain sequence and the light chain sequence are combined. The HCDR1 sequence, HCDR2 sequence, HCDR3 sequence, LC are defined by the system. DR1 sequence, LCDR2 sequence, and LCDR3 sequence, for example, sequence number 46, respectively. , further including 4, 5, 33, 14, and 15. In some other embodiments, the heavy chain sequence and The light chain sequence is defined by IMGT, consisting of the HCDR1 sequence, HCDR2 sequence, and HCDR 3 sequences, LCDR1 sequence, LCDR2 sequence, and LCDR3 sequence, for example, respectively, This further includes sequence numbers 43, 44, 45, 47, 37, and 15.

[0175] In other embodiments, the VH amino acid sequence and / or VL amino acid sequence are as shown in Table 1. The array is divided into 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, and 98%. They may be %, or 99%, identical. In other embodiments, the VH amino acid sequence and / or The VL amino acid sequence is an amino acid sequence with 1, 2, 3, 4, or 5 or fewer amino acid positions. They can be identical except for acid substitution. The VH and VL regions of the antibodies listed in Table 1 are large Antibodies having VH and VL regions with significant (i.e., 80% or more) identity are , the nucleic acids encoding sequence numbers 10 or 30, and sequence numbers 20 and 40, respectively. Molecular mutagenesis (e.g., site-directed mutagenesis or PCR-mediated mutagenesis) Subsequently, the encoded modified antibody is tested for its function using the functional assay described herein. This can be obtained by investigating retention.

[0176] In other embodiments, the full-length heavy chain amino acid sequence and / or the full-length light chain amino acid sequence are shown in the table. The sequence shown in 1 (for example, sequence numbers 11 and / or 21, or 31 and / Alternatively, 41), 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, They may be 98% or 99% identical. The full length of either sequence number 11 or 31. Heavy chain, and a full-length light chain of either sequence number 21 or 41 and a large (i.e., Antibodies having full-length heavy chains and full-length light chains with identity of 80% or more are such poly Mutagenesis of peptide-encoding nucleic acid molecules (e.g., site-directed mutagenesis or After PCR-mediated mutagenesis, use the functional assay described herein to code The modified antibody can be obtained by examining its ability to retain its function.

[0177] In one aspect of this specification, an isolated antibody comprising a heavy chain and a light chain or its functional antigen-binding properties A fragment is presented, and in this case, the heavy chain is selected from the group consisting of SEQ ID NOs: 11 and 31. The amino acid sequence and the a that is at least 80%, at least 90%, or at least 95% identical It contains a amino acid sequence; the light chain is an amino acid selected from the group consisting of SEQ ID NOs: 21 and 41. The sequence and amino acids that are at least 80%, at least 90%, or at least 95% identical. The sequence is included; the antibody is FXI and / or FXIa (e.g., human FXIa, rabbit F). It specifically binds to XIa and cynomolgus monkey FXIa). In one embodiment, isolated antibody Alternatively, the functional antigen-binding fragment comprises a heavy chain and a light chain, in which case the heavy chain is sequence The amino acid sequence of number 11, and at least 80%, at least 90%, or at least 9 It contains 5% identical amino acid sequences; the light chain has the amino acid sequence of SEQ ID NO: 21 and at least 8 The antibody contains 0%, at least 90%, or at least 95% identical amino acid sequences; FXI and / or FXIa (e.g., human FXIa, rabbit FXIa, and crab FXIa) It specifically binds to isal fXIa). In one embodiment, the isolated antibody or its functional antibody The primordial fragment includes a heavy chain and a light chain, in which case the heavy chain is the amino acid sequence of SEQ ID NO: 31. The row and the amino acid composition which is at least 80%, at least 90%, or at least 95% identical. The light chain includes the amino acid sequence of SEQ ID NO: 41, and at least 80%, at least 90% Containing %, or at least 95%, identical amino acid sequences; the antibody is FXI and / or FXIa (e.g., human FXIa, rabbit FXIa, and cynomolgus monkey FXIa) They bind heteromorphically. In certain aspects of the present invention, the heavy chain sequence and the light chain sequence are bonded to Kabat. The following are defined: HCDR1 sequence, HCDR2 sequence, HCDR3 sequence, LCDR1 sequence, LCDR2 sequence and LCDR3 sequence, for example, sequence numbers 3, 4, 5, and 13, respectively. , 14, and 15 further include. In certain other embodiments of the present invention, heavy chain arrangement and light chain The sequences are defined by Chothia: HCDR1 sequence, HCDR2 sequence, HCDR 3 sequences, LCDR1 sequence, LCDR2 sequence, and LCDR3 sequence, for example, respectively, The sequence numbers 6, 7, 8, 16, 17, and 18 are further included. In certain other embodiments, The chain sequence and light chain sequence are defined by a combination system, HCDR1 sequence, HCDR 2 sequences, HCDR3 sequence, LCDR1 sequence, LCDR2 sequence, and LCDR3 sequence, example For example, each further includes sequence numbers 46, 4, 5, 33, 14, and 15. In other embodiments, the heavy chain sequence and light chain sequence are defined by IMGT, HCDR1 sequence Column, HCDR2 sequence, HCDR3 sequence, LCDR1 sequence, LCDR2 sequence, and LCD R3 sequences, for example, sequence numbers 43, 44, 45, 47, 37, and 15 respectively It also includes.

[0178] In other embodiments, the nucleotide sequences of the full-length heavy chain and / or full-length light chain are shown in Table 1. The array being processed (for example, arrays 12 and / or 22, or 32 and / or 42) and 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% They can be % identical.

[0179] In other embodiments, the nucleotide sequences of the heavy chain variable region and / or the light chain variable region are: The sequences shown in Table 1 (for example, sequence numbers 10 and / or 20, or 30 and / Alternatively, 40) and 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% They are 99% identical.

[0180] As used herein, the percentage of identity between two sequences is the optimal value of the two sequences. Consider the number of gaps that need to be introduced for alignment and the length of each gap. A function of the number of identical positions shared by the array (i.e., identity %= identical positions) (Number / Total number of positions × 100). Comparison of arrays and identity percentage between two arrays. The decision can be achieved using a mathematical algorithm, as described in the non-restrictive example below. It is possible.

[0181] In addition, or alternatively, the protein sequences of the present invention are in reference to published databases. To perform a search, for example, as a "query sequence" to identify related sequences, It can also be used for this purpose. For example, such a search was conducted by Altschul et al., 1990 J. Mol. Biol. This should be done using the BLAST program (version 2.0) as specified in 215:403~10. It is possible.

[0182] Antibodies with Conservative Modifications In certain embodiments, the antibody of the present invention comprises a CDR1 sequence, a CDR2 sequence, and a CDR A heavy chain variable region containing 3 sequences, as well as CDR1, CDR2, and CDR3 sequences It has a light chain variable region including, in this case, one or more of these CDR sequences Based on the antibodies described herein or their conservative modifications, the amino acid sequences specified are The antibody possesses the desired functional properties of the FXIa-binding antibody of the present invention.

[0183] Therefore, the present invention relates to a heavy chain comprising the CDR1 sequence, the CDR2 sequence, and the CDR3 sequence. Variable region, and light chain variable region including CDR1 sequence, CDR2 sequence, and CDR3 sequence. An isolated antibody or its antigen-binding fragment comprising a region, wherein the heavy chain variable region is CDR1 amino The acid sequence was selected from the group consisting of SEQ ID NOs: 3 and 23, and their conservative modifications. The CDR2 amino acid sequences of the heavy chain variable region are those of SEQ ID NOs: 4 and 24, and their respective retention Selected from a group consisting of residual modifications, the CDR3 amino acid sequence of the heavy chain variable region is SEQ ID NO: 5 and 25, and selected from the group consisting of their conservative modifications, the CD of the light chain variable region. The R1 amino acid sequence consists of sequence numbers 13 and 33, and their conserved modifications. Selected from, if the CDR2 amino acid sequence of the light chain variable region is sequence numbers 14 and 34, Furthermore, selected from the group consisting of these conserved modifications, the CDR3 amino acid sequence of the light chain variable region. However, selected from the group consisting of sequence numbers 15 and 35, and their conservative modifications, L The present invention provides an antibody or its antigen-binding fragment that specifically binds to OX-1.

[0184] In one aspect of this specification, a heavy chain comprising the CDR1 sequence, the CDR2 sequence, and the CDR3 sequence is used. Variable region, and light chain variable region including CDR1 sequence, CDR2 sequence, and CDR3 sequence. An isolated antibody or its antigen-binding fragment comprising a region, wherein the heavy chain variable region is CDR1 amino The acid sequence is selected from the group consisting of amino acid sequences and their conserved modifications listed in Table 1. Furthermore, the CDR2 amino acid sequence of the heavy chain variable region is the same as the amino acid sequence listed in Table 1. Selected from a group consisting of conserved modifications; the CDR3 amino acid sequence of the heavy chain variable region is shown. Selected from the group consisting of amino acid sequences and their conserved modifications described in 1; light chain possible The CDR1 amino acid sequence of the variant region is the amino acid sequence listed in Table 1 and its conservation Selected from the group consisting of modifications; the CDR2 amino acid sequence of the light chain variable region is listed in Table 1. Selected from the group consisting of amino acid sequences and their conserved modifications; CD of the light chain variable region The R3 amino acid sequence consists of the amino acid sequences and their conserved modifications listed in Table 1. Selected from a group; the antibody or its antigen-binding fragment specifically binds to FXIa.

[0185] In other embodiments, the antibody of the present invention is optimized for expression in mammalian cells. It has a full-length heavy chain sequence and a full-length light chain sequence, and one or more of these sequences , amino acid sequences specified based on the antibodies described herein or their conservative modifications The antibody possesses the desired functional properties of the FXIa-binding antibody of the present invention. Therefore, the present invention provides a full-length heavy chain and a full-length light chain for expression in mammalian cells. Optimized isolated antibodies, the full-length heavy chain of which is sequence number 11 or 31, and their It has an amino acid sequence selected from the group of conservative modifications, and the full-length light chain is SEQ ID NO: 21 or 4 1, and having an amino acid sequence selected from the group of their conserved modifications, FXI and / or specific to FXIa (e.g., FXIa in humans, rabbits, and cynomolgus monkeys). Provides an antibody that binds to this substance.

[0186] The same epitope-binding antibody The present invention relates to the FXI-binding antibodies and / or FXIa-binding antibodies listed in Table 1. It provides antibodies that bind to the same epitopes as the body. Therefore, FXI and / or FX Ia binding assay (FXI and / or FXIa binding assay as described in the Examples section) In this case, the binding of the antibody to the other antibody of the present invention competes with other antibodies of the present invention (for example, the binding of the antibody to the other antibody of the present invention is the same). Binding to an epitope or overlapping epitope competitively inhibits in a statistically significant manner. Based on their ability to harm, further antibodies can be identified. The ability of the test antibody to inhibit binding to the FXI protein and / or FXIa protein. The force causes the test antibody to compete for binding to FXI and / or FXIa. It is possible to combine such antibodies, and according to a non-limiting theory, the FXI protein Whether the quality and / or FXIa protein is the same as or related to the competing antibody It can bind to epitopes of a series (for example, those that are structurally similar or spatially close). This is supported. In certain embodiments, the same FXI and / or antibody as the present invention Antibodies that bind to epitopes on FXIa are human monoclonal antibodies. Human monoclonal antibodies can be prepared and isolated as described herein. Cut.

[0187] As used herein, in the presence of equimolar concentrations of competing antibodies, the competing antibodies are as described in the present invention. The same FX as the antibody or antigen-binding fragment (e.g., NOV1401 or NOV1090) Binds to the I epitope and / or FXIa epitope, and binds to the antibody or antigen of the present invention. The binding of the sex fragment to FXI and / or FXIa exceeds 50% (e.g., 80%) If the antibody inhibits 85%, 90%, 95%, 98%, or 99%, then the antibody will bind to the target substance. "To be in competition" means, for example, by any of the methods well known to those skilled in the art, This can be determined in a combined assay.

[0188] The antibodies or antigen-binding fragments used herein are the competing antibodies or their antigens. The binding fragment is the same FXI epitope as the antibody or antigen-binding fragment of the present invention and / or ku is an FXIa epitope, or an overlapping FXI epitope and / or FX Insofar as it does not bind to the Ia epitope, the FXI antibody and / or FXI of the present invention a antibody or antigen-binding fragment (e.g., NOV1401 or NOV1090) and "competition" "Do not." The competitive antibodies or antigen-binding fragments used herein are (i) the present invention Steric blocking the binding of an antibody or antigen-binding fragment to its target (for example, before The competitive antibody is located near non-overlapping FXI epitopes and / or FXIa epitopes. By binding, the antibody or antigen-binding fragment of the present invention is physically prevented from binding to its target. (ii) different, non-overlapping FXI epitopes and / or It binds to the FXIa epitope, and the FXI antibody of the present invention and / or the protein This occurs when an FXIa antibody or antigen-binding fragment is formed in the absence of the conformational change. To prevent binding in that form, conformational changes are made to the FXI protein and / or It does not contain competitive antibodies or their antigen-binding fragments that induce the FXIa protein.

[0189] Modified antibodies and manipulated antibodies As a starting material for manipulating modified antibodies whose properties have been altered from the starting antibody, the following are shown herein. Using an antibody having one or more of the VH sequence and / or VL sequence, The antibody can be further prepared. The antibody has one or both variable regions (i.e.) , within VH and / or VL, for example, within one or more CDR areas, and / or Alternatively, modifying one or more residues within one or more framework regions. It can be manipulated more effectively. In addition, or alternatively, the antibody can modify residues within the constant region. For example, it can also be manipulated by modifying the effector function of the antibody. .

[0190] One type of variable region manipulation that can be performed is CDR grafting. The antibody is , primarily through amino acid residues located within the six heavy and light chain complementarity-determining regions (CDRs) It then interacts with the target antigen. For this reason, the amino acid sequence within the CDR is between individual antibodies. The diversity of the CDR sequence is greater than that of the sequence outside the CDR. The CDR sequence is the most antibody-antigen interaction Because this is one of the factors, the framework sequences are derived from different antibodies with different properties. By constructing an expression vector containing a CDR sequence derived from a rafted specific spontaneously occurring antibody, This makes it possible to express recombinant antibodies that mimic the characteristics of specific naturally occurring antibodies. (For example, 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; Win U.S. Patent No. 5,225,539 by ter, and U.S. Patent No. 5,225,539 by Queen et al. Patent No. 5,530,101; Patent No. 5,585,089; Patent No. 5,693 See also Specification No. 762 and Specification Nos. 6,180,370.

[0191] Therefore, another embodiment of the present invention is selected from the group consisting of Sequence IDs 3 and 23. Selected from the group consisting of CDR1 sequences, SEQ ID NOs: 4 and 24, which have the amino acid sequence. A selection from the group consisting of CDR2 sequences having amino acid sequences, SEQ ID NOs. 5 and 25. A heavy chain variable region containing each of the CDR3 sequences having a mino acid sequence, and Sequence ID No. 13 and CDR1 sequence having an amino acid sequence selected from a group of 33, SEQ ID NO: 14 and A CDR2 sequence having an amino acid sequence selected from a group of 34, and Sequence ID No. 1 Each of the CDR3 sequences consisting of amino acid sequences selected from the groups of 5 and 35 is This relates to an isolated antibody or its antigen-binding fragment containing a light chain variable region. Therefore, Such antibodies contain the VH CDR sequence and VL CDR sequence of the monoclonal antibody. They possess, but may contain different framework sequences derived from these antibodies.

[0192] Such framework sequences include published D sequences, which include germline antibody gene sequences. This can be obtained from the NA database or published references. For example, human heavy chain The germline DNA sequences of the light chain variable region genes are from the human germline sequence database. The "VBase" (available on the internet at mrc-cpe.cam.ac.uk / vbase) In addition to the above, the contents of each of them are expressly incorporated herein by reference, Kaba t, EA et al., 1991. Sequences of Proteins of Immunological Interest, 5th edition, US Department. rtment of Health and Human Services, NIH Publication No. 91-3242; Tomlinson, I. M. . et al., 1992 J. Mol. Biol. 227:776~798; and Cox, JPL et al., 1994 Eur. J Immunol It can also be found in 24:827-836.

[0193] Examples of framework sequences for use in antibodies of the present invention include selected antibodies of the present invention. Framework sequences used by the body, for example, used by the monoclonal antibody of the present invention The consensus sequence and / or framework sequence used are structurally similar to the framework These are the VH CDR1 sequence, VH CDR2 sequence, and VH CDR3 sequence. The sequences, as well as the VL CDR1 sequence, VL CDR2 sequence, and VL CDR3 sequence are The framework sequence is found within germline immunoglobulin genes from which it originates. It is also possible to graft the column into a framework region that has the same sequence, and the CDR sequence is Compared to germline sequences, framework regions containing one or more mutations It is also possible to graft into a domain. For example, in certain cases, within a framework domain It is considered beneficial to mutate residues to maintain or enhance the antigen-binding ability of antibodies. (For example, U.S. Patent No. 5,530,101 by Queen et al.; same) Specification No. 5,585,089; Specification No. 5,693,762; and Specification No. 6,18 See Specification No. 0,370. Antibodies and antigen-binding fragments described herein. Frameworks that can be used as a scaffolding to build upon are VH1A, VH1B, V This includes, but is not limited to, H3, Vk1, Vl2, and Vk2. In the art, Further frameworks are known, for example, the internet's vbase.mrc-cpe.cam. It can be found in the vBase database at ac.uk / index.php?&MMN_position=1:1 can.

[0194] Therefore, embodiments of the present invention are selected from the group consisting of Sequence IDs 9 and 29. Mino acid sequence, or within the framework region of such sequence, 1, 2, 3, 4 or Contains an amino acid sequence with five amino acid substitutions, deletions, or additions. An amino acid sequence that includes a heavy chain variable region and is selected from the group consisting of SEQ ID NOs: 19 or 39. , or within the framework region of such an array, 1, 2, 3, 4, or 5 locations Light chains having amino acid sequences with amino acid substitutions, amino acid deletions, or amino acid additions are possible. This relates to isolated FXIa-binding antibodies or antigen-binding fragments thereof, further comprising a variant region.

[0195] Another type of variable region modification is known as "affinity maturation" in the VH CDR1 region. Within the region, within the VH CDR2 region, and / or within the VH CDR3 region, and / or This is within the VL CDR1 region, the VL CDR2 region, and / or the VL CDR3 region. By mutating the amino acid residues within the molecule, the binding properties of one or more of the target antibody are altered. The goal is to improve sex (e.g., affinity). Site-directed mutagenesis or P Mutations can be introduced by performing CR-mediated mutagenesis, as described herein. As presented in the Examples section, in vitro assays or in vivo assays In this way, the effect on antibody binding or other functional properties of the target can be assessed. Conservative modifications (discussed above) can be introduced. Mutation occurs at the site of amino acid substitution. There may be additions, deletions, and moreover, typically within the CDR region. Modify one, two, three, four, or five or fewer residues.

[0196] Therefore, in another embodiment, the present invention is selected from the group consisting of Sequence IDs 3 and 23. The amino acid sequence, or compared to SEQ ID NOs: 3 and 23, 1, 2, 3, 4 or This refers to an amino acid sequence having five amino acid substitutions, amino acid deletions, or amino acid additions. Amino acid combinations selected from the group consisting of the VH CDR1 region, SEQ ID NOs: 4 and 24 Compared to the column, or sequence numbers 4 and 24, there are 1, 2, 3, 4, or 5 amino acids. VH CD having an amino acid sequence with acid substitution, amino acid deletion, or amino acid addition Amino acid sequence selected from the group consisting of the R2 region, SEQ ID NOs. 5 and 25, or sequence number Compared to numbers 5 and 25, there are 1, 2, 3, 4, or 5 amino acid substitutions, amino acids VH CDR3 region having an amino acid sequence with deletion or amino acid addition, sequence number An amino acid sequence selected from the group consisting of numbers 13 and 33, or SEQ ID NOs. 13 and 3 Compared to 3, there are 1, 2, 3, 4, or 5 amino acid substitutions, amino acid deletions, or The VL CDR1 region has an amino acid sequence with amino acid addition, SEQ ID NO: 14 and An amino acid sequence selected from a group of 34, or compared with SEQ ID NOs: 14 and 34 , 1, 2, 3, 4, or 5 amino acid substitutions, amino acid deletions, or amino acid additions The VL CDR2 region having an amino acid sequence containing the addition, and SEQ ID NOs. 15 and 35 An amino acid sequence selected from the group consisting of, or compared with SEQ ID NOs: 15 and 35, , 2, 3, 4, or 5 amino acid substitutions, amino acid deletions, or amino acid additions Isolation FX consisting of a heavy chain variable region having a VL CDR3 region having an amino acid sequence This invention provides an Ia-binding antibody or an antigen-binding fragment thereof.

[0197] Therefore, in another embodiment, the present invention selects from the group consisting of Sequence IDs 6 and 26. The amino acid sequence, or compared to SEQ ID NOs. 6 and 26, is 1, 2, 3, 4 or This refers to an amino acid sequence having five amino acid substitutions, amino acid deletions, or amino acid additions. Amino acid combinations selected from the group consisting of the VH CDR1 region, SEQ ID NOs: 7 and 27 Compared to the column, or sequence numbers 7 and 27, there are 1, 2, 3, 4, or 5 amino acids. VH CD having an amino acid sequence with acid substitution, amino acid deletion, or amino acid addition The R2 region, and an amino acid sequence selected from the group consisting of SEQ ID NOs: 8 and 28, Compared to Sequence IDs 8 and 28, it has 1, 2, 3, 4, or 5 amino acid substitutions. VH CDR3 region having an amino acid sequence with amino acid deletion or amino acid addition A heavy chain variable region having and an amino acid sequence selected from the group consisting of SEQ ID NOs: 16 and 36 Compared to the row, or sequence numbers 16 and 36, there are 1, 2, 3, 4, or 5 meshes. VL C having an amino acid sequence with an acid substitution, amino acid deletion, or amino acid addition. An amino acid sequence selected from the group consisting of the DR1 region, SEQ ID NOs: 17 and 37, or Compared to rows 17 and 37, there are 1, 2, 3, 4, or 5 amino acid substitutions. VL CDR2 region having an amino acid sequence with amino acid deficiency or amino acid addition, Also, an amino acid sequence or sequence number selected from the group consisting of SEQ ID NOs. 18 and 38. Compared to numbers 18 and 38, there are 1, 2, 3, 4, or 5 amino acid substitutions. It has a VL CDR3 region with an amino acid sequence that is either oxygen-deficient or has amino acid addition. This provides isolated FXIa-binding antibodies or antigen-binding fragments thereof, consisting of a light chain variable region. do.

[0198] Grafting antigen-binding domains onto alternative frameworks or scaffolds The resulting polypeptide has at least one bond that specifically binds to FXIa. Insofar as it includes the compatibility region, a wide variety of antibody / immunoglobulin frameworks or A scaffold can be used. Such a framework or scaffold is used in human immunoglobulins. It comprises five main idiotypes of phosphorus or fragments thereof, preferably with humanized aspects It contains immunoglobulins from other animal species. In this respect, it is identified in camelids. Single heavy chain antibodies, such as single heavy chain antibodies, are particularly targeted. Those skilled in the art can develop novel frameworks. Scrap, scaffolding, and fragments continue to be discovered and developed.

[0199] In one embodiment, the present invention uses a non-immunoglobulin scaffold capable of grafting the CDR of the present invention. This relates to producing non-immunoglobulin-based antibodies. They target FXI Insofar as it includes a binding domain specific to proteins and / or FXIa proteins , utilizing publicly known or future non-immunoglobulin frameworks and non-immunoglobulin scaffolds This is possible. Known non-immunoglobulin frameworks and non-immunoglobulin scaffolds fibronectin (Compound Therapeutics, Inc., Wa (Molecular Partners AG, Zu) rich, Switzerland), domain antibody (Domantis, Ltd., C ambridge, MA; and Ablynx nv, Zwijnaarde, Belgium ium), lipocalin (Pieris Proteolab AG, Freising, Germany), small molecule modular immunotherapy (Trubion Pharmaceut icals Inc., Seattle, WA), Maxibody (Avidia, Inc.) ., Mountain View, CA), Protein A (Affibody AG, S weden), and affilin (gamma-crystallin or ubiquitin) (Scil This includes, but is not limited to, Proteins GmbH (Halle, Germany). I can't.

[0200] The fibronectin scaffold consists of type III fibronectin domains (for example, type III fibronectin domains). Based on the 10th module (10 Fn3 domain) of bronnectin. Type III fibr Ronectin domains pack themselves together to form the core of the protein. Furthermore, it contains loops that connect the beta chains to each other and are exposed to the solvent (CDR and (Similar), having seven or eight beta chains distributed between two beta sheets. Each edge of the beta sheet sandwich has at least three such loops. The edge is the protein boundary perpendicular to the direction of the beta chain (US6,818, (See 418). The overall fold is antigen recognition within camel and llama IgG. It is closely related to the fold of the heavy chain variable region, which is the smallest functional antibody fragment containing the entire unit. However, these fibronectin-based scaffolds are not immunoglobulins. Therefore, non-immunoglobulin antibodies have properties and affinity that are related to the properties and affinity of antibodies. They mimic antigen-binding properties similar to those of tee. These scaffolds are used in vivo to inhibit antigen binding. Randomness of loops in vitro, similar to the process of affinity maturation in the body. These fibronects can be used in transformation strategies and shuffling strategies. The base molecule is cloning the molecular loop region using standard cloning methods according to the present invention. It can be replaced with DR and used as scaffolding.

[0201] Ankyrin technology involves using proteins with repeat modules derived from ankyrin to create different proteins. This is based on the idea of ​​using it as a scaffold that has a variable region that can be used for binding to a target. The NKIRIN Repeat Module consists of two antiparallel α-helix and β-helix circuits. It is a polypeptide consisting of 33 amino acids. The binding of the variable region is via ribosomal decomposition. Using a spray bottle will optimize most of the process.

[0202] Abimers are derived from naturally occurring A-domain-containing proteins such as LRP-1. Domains are primarily used in protein-protein interactions, and in humans, there are over 250 protein domains. The molecule is structurally based on the A domain. The avimer is linked via an amino acid linker. It consists of numerous (2 to 10) different "A domain" monomers that can bind to target antigens. Abimar, for example, in U.S. Patent Application Publication No. 20040175756; and the same publication No. 200 Specification No. 50053973; Specification No. 20050048512; and Specification No. 2006 It can be created using the method described in Specification No. 0008844.

[0203] Affinity ligands, or affibodies, are IgG-binding domains of protein A. A simple, low-molecular-weight protein consisting of a 3-helix bundle based on one of the scaffolds. It is a quality. Protein A is produced by the bacterium Staphylococcus aureus. These are surface proteins of which are derived. Of these scaffold domains, 13 are composed of numerous Randomized 58A to create an affibody library with Gand variants. Composed of amino acids (see, for example, US5,831,012). Affibody molecules It mimics antibodies, and compared to the molecular weight of an antibody which is 150 kDa, it has a molecular weight of 6 kDa. Despite its small size, the binding site of the affibody molecule is the same as the binding site of the antibody. It appears that way.

[0204] Antikalin is a compound developed by Pieris ProteoLab AG. It is a substance. Antikarin is typically a chemically sensitive compound or an insoluble compound. Lipocalin is a broad group of robust, small-molecule proteins involved in biological transport or storage. It is derived from phosphorus. Some natural lipocalins occur in human tissues or human body fluids. Protein architecture involves a highly variable loop on top of a rigid framework. It is reminiscent of immunoglobulins. However, in contrast to antibodies or recombinant fragments thereof, Pokarin has 160-180 ami, which is just slightly larger than a single immunoglobulin domain. It consists of a single polypeptide chain with no acid residues. It has four loops that form a binding pocket. The set exhibits remarkable structural plasticity and allows for various side chains. Therefore, different shapes To recognize the specified target molecule with high affinity and specificity, the binding site It can be reformed through a unique process. By inducing mutations in a set of four loops. To develop antikalin, one of the proteins in the lipocalin family is The bilin-binding protein (BBP) of the large white butterfly (Pieris Brassicae) was used. One example of a patent application describing antikarin is a PCT international publication. It is in pamphlet number 199916873.

[0205] Affilin molecules are characterized by their specific affinity for proteins and small molecules. It is a small non-immunoglobulin protein designed for this purpose. The novel affilin molecule is Each of them is very different from two libraries based on human-derived scaffold proteins. It can be quickly selected. Affilin molecules are against immunoglobulin proteins. It shows no structural homology whatsoever. Currently, two affilin scaffolds are being used, and they One of them is gamma crystallin, a structural protein of the human lens, and the other is These are "ubiquitin" superfamily proteins. All of the human scaffolds are very small. Furthermore, it exhibits high thermal stability and is almost resistant to pH changes and denaturing agents. The stability is mainly due to the unfolding of the protein's beta-sheet structure. Examples of derived proteins are described in WO200104144, and are referred to as "ubiquitous An example of a "Chin-like" protein is described in WO2004106368.

[0206] Protein epitope mimes (PEMs) are major proteins involved in protein-protein interactions. A medium-sized secondary structure that mimics the beta-hairpin secondary structure of proteins. It is a cyclic peptide-like molecule (molecular weight: 1-2 kDa).

[0207] This invention provides a fully human antibody that specifically binds to the FXIa protein. Compared to human or humanized antibodies, the human FXIa-binding antibody of the present invention is administered to a human subject. The antigenicity is further reduced when this occurs.

[0208] camelid animal antibodies Llama species (alpaca (Lama paccos), llama (Lama glama), and vicuña (Lama vi) New World members such as cugna, camels and dromedary camels (F Octopus-humped camel (Camelus bactrianus) and dromedary camel (Calelus dromaderius) Antibody proteins obtained from members of the family differ in size, structural complexity, and hygroscopicity. It is characterized in terms of antigenicity against the target. This family found in nature Certain IgG antibodies derived from mammals lack a light chain and are therefore derived from other animals. In the case of antibodies, the typical quaternary structure is a four-chain structure with two heavy chains and two light chains. They are structurally different. PCT / EP93 / 02214 (published on March 3, 1994) Please refer to the internationally published pamphlet No. 94 / 04678.

[0209] The region of camelid antibodies, identified as a small, single variable domain known as VHH, is Because it yields small molecules that have high affinity for the target, " This results in low molecular weight antibody-derived proteins known as "Petulidae nanobodies." It can be obtained through genetic engineering. (U.S. Patent No. 5,7, obtained June 2, 1998) See issues 59,808. Also, Stijlemans, B. et al., 2004 J Biol Chem 279:1256. ~1261; Dumoulin, M. et al., 2003 Nature 424:783~788; Pleschberger, M. et al., 2003 Bioco njugate Chem 14:440~448; Cortez-Retamozo, V. et al., 2002 Int J Cancer 89:456~62; See also Lauwereys, M. et al., 1998 EMBO J 17:3512~3520. Camelid antibodies And the manipulation libraries for antibody fragments are, for example, Ablynx, Ghent, Belgi It is commercially available from UM. Like other non-human antibodies, it is a camelid antibody with amino acids. The sequence can be modified by recombination to obtain a sequence that more closely resembles the human sequence. In other words, nanobodies can be "humanized". Therefore, the relationship with humans The naturally low antigenicity of antibodies from pipefish can be further reduced.

[0210] The molecular weight of camelid nanobodies is about one-tenth that of human IgG molecules, The physical diameter of the protein is only a few nanometers. One consequence of its small size is Large antibody proteins bind to antigenic sites that are functionally invisible to camelids. The ability of the nanobody, namely the camelid nanobody, is to utilize classical immunological techniques. It is useful as a reagent for detecting antigens that remain hidden in other forms, and is possible. It is useful as a therapeutic agent. Therefore, another consequence of its small size is that it is easy to use. Murid nanobodies target specific parts within grooves or narrow crevices of target proteins. As a result of binding to the site, it is possible to inhibit the target protein, and therefore, In its ability to more closely resemble the function of classical low molecular weight drugs than the function of classical antibodies. It can be used.

[0211] Furthermore, its low molecular weight and compact size give it extremely high thermal stability, even at extreme pH levels. Furthermore, camelid nanobodies that are stable against proteolytic digestion and have low antigenicity This results in... Another consequence is that camelid nanobodies move from the circulatory system to the tissues. It can easily move, cross the blood-brain barrier, and treat damage affecting nerve tissue. Furthermore, nanobodies can facilitate drug delivery across the blood-brain barrier. (2004) Please refer to U.S. Patent Application No. 20040161738, published on August 19. By combining these characteristics with low antigenicity to humans, a large Therapeutic potential is indicated. Furthermore, these molecules are found in prokaryotes such as Escherichia coli (E. coli). It can be fully expressed within cells and is expressed as a fusion protein with bacteriophages. It is expressive and functional.

[0212] Therefore, a feature of the present invention is high affinity for FXI and / or FXIa. A camelid antibody or camelid nanobody having a certain species. In this embodiment, camelid animal antibodies or camelid animal nanobodies are naturally occurring in camelids. This involves producing antibodies in animals, i.e., using the techniques described herein for other antibodies. After immunization with FXI and / or FXIa or its peptide fragments, Produced by gullids. Alternatively, FXI and / or FXIa-binding camelids. The material nanobody is manipulated, i.e., in the examples of this specification. The FXI and / or FXIa described as targets, and / or Using a panning procedure involving domains and / or peptide fragments, appropriately mutate Selection from a phage library presenting induced camelid nanobody proteins It is created by selection. The manipulated nanobody is further genetically engineered to create a recipe. The half-life of the target substance can also be customized to range from 45 minutes to 2 weeks. In specific embodiments, camelid antibody or camelid nanobody is used, for example, As described in PCT / EP93 / 02214, the heavy chain of the human antibody of the present invention Alternatively, the CDR sequence of the light chain is converted into a nanobody or single-domain antibody framework sequence. Obtained by rafting.

[0213] Dispecific molecules and polyvalent antibodies In another embodiment, the present invention relates to the FXI-binding antibody and / or FXIa-binding antibody of the present invention. The present invention features a bispecific or polyspecific molecule containing the body or fragments thereof. Alternatively, its antigen-binding region can bind to at least two different binding sites or target molecules. It can also be derivatized to create two specific molecules, or another functional molecule, for example, another Linked to peptides or proteins (e.g., another antibody or ligand against the receptor). It can also bind. The antibody of the present invention actually has three or more different binding sites and / or It is also possible to derivatize them to create highly specific molecules that bind to target molecules, and the other two are different. It can also be linked to the functional molecules above, and such polyspecific molecules are also used herein. The term "two-specific molecules" used in this invention is intended to encompass them. To create isomers, the antibody of the present invention is used such that a bispecific molecule is obtained as a result. One or more other binding agents, such as another antibody, antibody fragment, peptide, or binding mimetic. Functionally linking to molecules (e.g., chemical coupling, gene fusion, non-covalent bonding) It can be done by combination or in other ways.

[0214] Therefore, the present invention relates to at least one of the following for FXI and / or FXIa A two-specificity component comprising a first binding specificity and a second binding specificity for a second target epitope. Including offspring. For example, the second target epitope is different from the first target epitope, FXI It is a calligraphy / or another epitope of FXIa.

[0215] In addition, in the present invention, where the bispecific molecule is polyspecific, the molecule targets the first target epitope. In addition to the second target epitope, a third binding specificity may also be included.

[0216] In one embodiment, the two specific molecules of the present invention have binding specificity as, for example, Fab, Fa At least one antibody or It contains antibody fragments. Antibodies also include light chain or heavy chain dimers, or Ladner et al., USA Fv or single-chain construct as described in Japanese Patent No. 4,946,778 It could be any smallest fragment.

[0217] A diabody is a rib that is too short to allow pairing between two domains on the same chain. The VH and VL domains, linked by a linker, are generated on a single polypeptide chain. It is a divalent, bispecific molecule. The VH domain and VL domain are on the other side of the chain. It pairs with a complementary domain, thereby creating two antigen-binding sites (for example, Hol Iger et al., 1993 Proc. Natl. Acad. Sci. USA 90:6444~6448; Poljak et al., 1994 Structure See 2:1121~1123). The diamond bodies are VHA-VLB and VHB-VLA Structure (VH-VL stereoconfiguration), or VLA-VHB and VLB-VHA structure (VL- This is created by expressing two polypeptide chains with a VH (Very High-Hens) configuration within the same cell. It is possible. Most of the diabolic bodies can be expressed in a soluble form within bacteria. A single-chain diabody (scDb) consists of two diabody-forming polypeptide chains, approximately 1 It is constructed by linking with a 5-amino acid linker (Holliger and Winter, 1997). Cancer Immunol. Immunother., 45(3~4):128~30; Wu et al., 1996 Immunotechnology, 2 (See (1):21-36). scDb is expressed in a soluble and active monomeric form within bacteria. (Holliger and Winter, 1997 Cancer Immunol. Immunother., 45( 34):128~30; Wu et al., 1996 Immunotechnology, 2(1):21~36; Pluckthun and Pack, 1 997 Immunotechnology, 3(2):83~105; Ridgway et al., 1996 Protein Eng., 9(7):617~ (See 21). By fusing the Diabody with Fc, a "Diabody" is created. This is possible (see Lu et al., 2004 J. Biol. Chem., 279(4):2856~65).

[0218] Other antibodies that can be used within the two-specific molecules of the present invention include mouse monoclonal antibodies and chimeric antibodies. These are monoclonal antibodies and humanized monoclonal antibodies.

[0219] Two specific molecules are obtained by using methods known in the art to determine the binding specificity of their constituent components. It can be prepared by denjutting. For example, each binding characteristic of two specific molecules The opposite sex can be created individually and then conjugated with each other. If the substance is a protein or peptide, various coupling agents or crosslinking agents can be used together. It can be used in bound conjugations. Examples of crosslinking agents include protein A, ca Rubodiimide, N-succinimidyl-S-acetyl-thioacetate (SATA), 5 ,5'-Dithiobis(2-nitrobenzoic acid)(DTNB), o-Phenylenedimaleimide (oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (S PDP), and sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane Contains -1-carboxylate (sulfo-SMCC) (e.g., Karpovsky et al., 1984 J. E.) See xp. Med. 160:1686; Liu, MA et al., 1985 Proc. Natl. Acad. Sci. USA 82:8648. (I want to). Other methods include Paulus, 1985 Behring Ins. Mitt. No. 78, 118-132; Brennan et al., 1 985 Science 229:81~83; and Glennie et al., 1987 J. Immunol. 139:2367~2375 The method described is included. The conjugate is SATA and sulfo-SMCC. Both are from Pierce Chemical Co. (Rockford, IL) It is available.

[0220] If the binding specificity is that of an antibody, they bind the C-terminal hinge region of the two heavy chains. They can be conjugated by drilling them together. In a particular embodiment, Prior to conjugation, an odd number of sulfhydryl residues, for example, one sulfhydryl Modify the hinge region to include residues.

[0221] Alternatively, both binding specificities can be encoded within the same vector and expressed within the same host cell. It can be prepared and assembled. This method allows two specific molecules to be mAb × mAb Fusion protein, mAb×Fab fusion protein, Fab×F(ab')2 fusion protein This is particularly useful when it is a quality or ligand × Fab fusion protein. The specificity molecule may be a single-chain molecule containing one single-chain antibody and a binding determinant, or two binding molecules. In some cases, the molecule may be a single-chain bispecific molecule containing a synthesis determinant. A bispecific molecule is a single-chain molecule containing at least two single chains. It may contain molecules. For a method of preparing two specific molecules, see, for example, U.S. Patent No. 5,26 U.S. Patent No. 0,203; U.S. Patent No. 5,455,030; U.S. Patent No. 4,881, U.S. Patent No. 175; U.S. Patent No. 5,132,405; U.S. Patent No. 5,091,51 U.S. Patent No. 3; U.S. Patent No. 5,476,786; U.S. Patent No. 5,013,653 Specification; U.S. Patent No. 5,258,498; and U.S. Patent No. 5,482,858 It is described in the detailed specifications.

[0222] The binding of bispecific molecules to these specific targets is, for example, used in enzyme immunoassays. (ELISA), radioimmunoassay (REA), FACS analysis, bioassay (examples) For example, this can be confirmed by a growth inhibition assay or a Western blot assay. In each of these assays, a labeled reagent specific to the target complex is generally used. For example, by using an antibody, the presence of a specific protein-antibody complex can be identified. It is detected.

[0223] In another embodiment, the present invention relates to at least two identical antibodies of the present invention that bind to FXIa. The present invention provides a polyvalent compound comprising either one or a different antigen-binding moiety. via protein fusion, covalent linkage, or non-covalent linkage, together They can be linked. Alternatively, linking methods for dispecific molecules are also described. Tetravalentization The compound is, for example, an antibody of the present invention, and an antibody that binds to the constant region of the antibody of the present invention, for example, F This can be obtained by bridging with the c region or hinge region.

[0224] For example, regarding trimerization domains, see the patent EP10122 by Borean. It is described in 80B1. For pentamerization modules, for example, PCT / E This is described on page 97 / 05897.

[0225] Antibodies with extended half-life This invention specifically provides an extension of the half-life in vivo of the FXIa protein. Provides an antibody to bind to.

[0226] Many factors can affect the half-life of a protein in vivo. For example... For example, filtration in the kidneys, metabolism in the liver, and protein-degrading enzymes (proteases) Degradation and immunogenic response (e.g., neutralization of proteins by antibodies and macrophages) (Uptake by dendritic and dendritic cells). Various strategies are used to extend the half-life of the antibody of the present invention. It is possible. For example, polyethylene glycol (PEG), reCODE PEG Antibody scaffold, polysialic acid (PSA), hydroxyethyl starch (HES), albumin Through chemical linkage with a ligand that binds to IgG, and a carbohydrate shield, albumin, IgG , FcRn, and proteins that bind to serum proteins such as transferrin Through gene fusion, nanobodies, Fab, DARPin, avimers, aphibodies, and It couples with other binding sites that bind to serum proteins, such as antikalin (genes). By means of (either physically or chemically), rPEG, albumin, albumin domain, album By gene fusion with mine-binding proteins and Fc, or by nanocarriers, sustained-release formulations, Alternatively, the half-life of the antibody of the present invention can be extended by incorporating it into a medical device. .

[0227] To prolong the circulation of antibodies in serum in vivo, high molecular weight PEG, etc. Site-specific conjugation of the active polymer molecule to the N-terminus or C-terminus of PEG or the antibody. Via gating, or via the epsilon-amino group present on lysine residues, To antibodies or fragments thereof, with or without functional linkers. It can be attached to. To PEGylate an antibody, the antibody or a fragment of it is attached to polyethylene. PEG, such as a reactive ester or aldehyde derivative of glycol (PEG), and one Alternatively, the reaction is typically carried out under conditions in which multiple PEG groups are attached to the antibody or antibody fragment. PEGylation involves the interaction of reactive PEG molecules (or similar reactive water-soluble polymers) with each other. This can be carried out by acylation or alkylation reactions. The term "polyethylene glycol" refers to mono(C1-C10) alkoxy-polyethylene Polyethylene glycol or aryloxy polyethylene glycol or polyethylene glycol PEG, which is used to derivatize other proteins such as recall-maleimide It is intended to encompass any of the forms. In certain embodiments, P Antibodies that undergo EG conversion are deglycosylating antibodies. Linear polymers or branched polymers. Derivatization is used which derivatization results in minimal loss of biological activity. It is likely that the degree of conjugation will be closely determined by SDS-PAGE and mass spectrometry. By monitoring this, we can confirm the proper conjugation of the PEG molecule to the antibody. Yes, it is possible. Unreacted PEG can be removed by size exclusion chromatography or ion exchange chromatography. PEG can be separated from antibody-PEG conjugates by matrixing. Derivatized antibodies are prepared using methods well known to those skilled in the art, for example, as described herein. The assay can be used to investigate binding activity and in vivo efficacy. Yes, it is possible. In this field, methods for PEGylating proteins are known, and the antibody of the present invention is one such method. It can be applied to EP0154316 by Nishimura et al. See also EP0401384 by Ishikawa et al.

[0228] Other modified PEGylation techniques include tRNA synthetase and tRNA-containing reconstitution systems. Through this process, chemically identified side chains are incorporated into biosynthetic proteins, a reconstitution chemical direct method. reconstituting chemically orthogonal directed engineering ( Includes ReCODE PEG). This technology involves over 30 new amino acids in E. coli. Incorporation into biosynthetic proteins within E. coli cells, yeast cells, and mammalian cells. It is possible. tRNA can move non-natural amino acids to any position where an amber codon is located. It incorporates the stop amber codon and the incorporation signal of a chemically identified amino acid. It is converted into a codon for transmission.

[0229] Recombinant PEGylation technology (rPEG) can also be used to extend the serum half-life. This technology uses an unstructured protein tail of 300-600 amino acids to create an existing pharmaceutical-grade protein tail. This involves gene fusion with protein. Since the molecular weight is about 15 times its actual molecular weight, the serum half-life of a protein is significantly different. It increases. In contrast to conventional PEGylation, which requires chemical conjugation and repurification. As a result, the manufacturing process is greatly simplified, and the product is homogeneous.

[0230] Polysialylation uses polysialic acid (PSA), a natural polymer, to activate longevity Another technology that extends life and improves the stability of therapeutic peptides and therapeutic proteins. PSA is a polymer of sialic acid (sugar). It is used in proteins and therapeutic peptides. When used for drug delivery, polysialic acid provides a protective microenvironment during conjugation. This results in extending the active lifespan of therapeutic proteins in circulation, which in turn affects the immune system. To prevent recognition. PSA polymers are found naturally in the human body. SA has evolved over millions of years to coat those walls with PSA. It was adopted by the species of bacteria. Subsequently, these naturally polysiallylated bacteria were molecular Through imitation, it became possible to disable the body's internal defense systems. Nature's ultimate stealth technology. PSA can be easily obtained in large quantities from such bacteria, and with specific physical characteristics. It can be produced in this way. Bacterial PSA is chemically identical to PSA in the human body. Furthermore, even when coupled to a protein, it remains completely non-immunogenic.

[0231] Another technique involves the use of hydroxyethyl starch ("HES") derivatives linked to antibodies. It contains HES, a modified natural polymer derived from waxy corn starch. It is present and can be metabolized by enzymes in the body. HES solution is typically used to replenish blood volume and improve blood circulation. It is administered to improve the ology properties. HES conversion of the antibody increases the stability of the molecule. In addition to this, reducing renal clearance also makes it possible to extend the circulating half-life. This results in increased biological activity. Different parameters such as the molecular weight of HES are used. By modifying them, a wide range of HES antibody conjugates can be customized. can.

[0232] Antibodies with an extended half-life in vivo also exhibit one or more amino acid modifications. (i.e., substitution, insertion, or deletion) of the IgG constant domain or its FcRn binding Even when introduced into a sexual fragment (preferably an Fc domain fragment or a hinged Fc domain fragment), It can be extracted. For example, in the International Patent Publication No. 98 / 23289 pamphlet, International Patent Pamphlet No. 97 / 34631; and U.S. Patent No. 6,277,375 Please refer to the book.

[0233] Furthermore, to make antibodies or antibody fragments more stable in vivo, To extend the half-life in vivo, antibodies are used in albumin (e.g., human blood It can also be conjugated to clear albumin (HSA). In this technology, the technique This is well known, for example, in the pamphlet of International Patent Publication No. 93 / 15199, and the same No. 93 / 1 Brochure No. 5200, and Brochure No. 01 / 77137; and European Patent Please refer to Specification No. 413,622. In addition, the context of the bispecific antibodies described above. So, the specificity of an antibody is that one binding domain of the antibody binds to FXIa, The second binding domain of the antibody is configured to bind to serum albumin, preferably HSA. You can also design it.

[0234] Strategies for extending the half-life are desired when the half-life is extended in vivo. Nanobodies, fibronectin-based binders, and other antibodies or protein odors It is particularly useful.

[0235] Antibody conjugate The present invention relates to creating fusion proteins using heterologous proteins or heterologous polypeptides. to (or fragments thereof, preferably at least 10, at least 20, at least 30) , at least 40, at least 50, at least 60, at least 70, at least 80 (to polypeptides of at least 90, or at least 100 amino acids), by recombinant To fuse or chemically conjugate (covalent conjugation) (Including both covalent and non-covalent conjugations), specifically binds to the FXIa protein. The present invention provides antibodies or fragments thereof that can be combined. In particular, the present invention provides antibodies described herein. Antigen-binding fragments (e.g., Fab fragment, Fd fragment, Fv fragment, F(ab)2 fragment, VH) Domain (VH CDR, VL domain, or VL CDR), and heterogeneous protein, heterogeneous The present invention provides a fusion protein comprising a species polypeptide or a heterologous peptide. This involves fusing a protein, polypeptide, or peptide with an antibody or antibody fragment. Methods for causing denjutation are known. For example, U.S. Patent No. 5,336,603 5,622,929, 5,359,046, 5,34 Specification No. 9,053, Specification No. 5,447,851, and Specification No. 5,112,946 Specification No.; European Patent No. 307,434 and EP367,166; International Patent Publication No. 96 / 04388 and No. 91 / 06570 pamphlets Ashkenazi et al., 1991, Proc. Natl. Acad. Sci. USA 88:10535~10539; Zheng et al., 19 95, J. Immunol. 154:5590~5600; and Vil et al., 1992, Proc. Natl. Acad. Sci. USA 89 Please refer to 11337~11341.

[0236] Further fusion proteins are involved in gene shuffling, motif shuffling, and exonescence. Shuffling, and / or codon shuffling (collectively, "DNA shuffling") It can be produced through the technique (named). By using DNA shuffling, the present invention Modify the activity of antibodies or fragments thereof (e.g., high affinity, rapid dissociation) This can be an antibody with a low degree of efficacy or a fragment thereof. Generally, U.S. 5,605,7 Specification No. 93, Specification No. 5,811,238, Specification No. 5,830,721, Specification No. 5,830,721, Patent No. 5,834,252 and Patent No. 5,837,458; Patent et al., 19 97, 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 Bla SCO, 1998, Biotechniques 24(2):308~313 (Each of these patents and publications is referenced by...) See (which are incorporated into this specification in their entirety) for reference. Antibodies or Those fragments, or the antibodies that encode them, or those fragments, were subjected to errors before recombination. Random mutagenesis by prone PCR, random nucleotide insertion, or other methods It can be modified by applying the law. Anti-binding agent that specifically binds to the FXIa protein. A polynucleotide that codes for a body or a fragment thereof is one or more heterologous molecules. Alternatively, it can be rearranged using multiple components, motifs, segments, parts, domains, or fragments. .

[0237] Furthermore, antibodies or fragments thereof can be linked to marker sequences such as peptides to facilitate purification. They can also be fused with them. In a preferred embodiment, the amino acid sequence of the marker is one of them. Among the many commercially available options, pQE vectors (QIAGEN, Inc.) are particularly noteworthy. (9259 Eton Avenue, Chatsworth, CA 91311) The tag provided is a hexahistidine peptide (SEQ ID NO: 48). As described in z et al., 1989, Proc. Natl. Acad. Sci. USA 86:821~824, for example, For example, hexahistidine (SEQ ID NO: 48) allows for the simple purification of fusion proteins. Other peptide tags useful for purification are derived from influenza hemagglutinin protein. Hemagglutinin ("HA") corresponding to the epitope (Wilson et al., 1984, Cell 37:767) This includes, but is not limited to, the tags "g" and "flag".

[0238] In other embodiments, the antibodies or fragments thereof of the present invention are used in diagnostic agents or detectable agents. To conjugate with such antibodies, the onset, development, progression, and The severity and / or condition may be part of a clinical testing procedure, such as determining the effectiveness of a particular treatment. This can be useful for monitoring or prognosis diagnosis. Such diagnosis and detection are Antibodies were used to test for horseradish peroxidase, alkaline phosphatase, and beta-galactosidase. These include, but are not limited to, a variety of enzymes such as acetylcholinesterase. ;Streptavidin / biotin and avidin / biotin, etc., but not limited to these. Non-existent prosthetic group; umbelliferone, fluorescein, fluorescein isothiocyanate Rhodamine, dichlorotriazinylamine fluorescein, dansilchloride, and Examples of fluorescent materials include phycoerythrin, but are not limited to these; luminol, for example. Luminescent materials, but not limited to these; luciferase, luciferin, and aequorin. Bioluminescent materials, but not limited to these; iodine (131I, 125I, 123I) , and 121I), carbon (14C), sulfur (35S), tritium (3H), indiu M (115In, 113In, 112In, and 111In), Technetium (99T c) Thallium (20¹Tl), Gallium (68Ga, 67Ga), Palladium (10³ Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), 153 Sm, 177Lu, 159Gd, 149Pm, 140La, 175Yb, 166Ho, 9 0Y, 47Sc, 186Re, 188Re, 142Pr, 105Rh, 97Ru, 68G e, 57Co, 65Zn, 85Sr, 32P, 153Gd, 169Yb, 51Cr, 54 Examples of emission molecules include Mn, 75Se, 113Sn, and 117Tin, but are not limited to these. Electromagnetic materials; as well as positron-emitting metals and non-radioactive materials using various positron emission tomography techniques. It couples to detectable substances containing, but not limited to, paramagnetic metal ions. This can be achieved by doing so.

[0239] The present invention also includes the use of antibodies or fragments thereof conjugated into therapeutic portions. It includes antibodies or fragments thereof, cytotoxins, for example, cell proliferation inhibitors or cytotoxic agents. , the therapeutic agent or radioactive metal ions, for example, alpha emitters, to the therapeutic part of the condyloma. It can be used to negate any drug that is harmful to cells. Cytotoxins or cytotoxic agents can negate any drug that is harmful to cells. include.

[0240] Furthermore, antibodies or fragments of antibodies are therapeutic parts or drugs that modify a given biological response. It can also be conjugated into parts. The therapeutic or drug part is classically chemical It shall not be considered limited to therapeutic agents. For example, the drug portion may be a desired biological agent. It may be a protein, peptide, or polypeptide that possesses specific activity. Proteins include, for example, abrin, lysine A, Pseudomonas exotoxins, cholera toxin, and These include toxins such as diphtheria toxin; tumor necrosis factor, α-interferon, β-interferon Ron, nerve growth factor, platelet-derived growth factor, tissue plasminogen activator, apotosis Protein-modifying agents such as lymphokines, anti-angiogenic agents; or, for example, other bio-response modifiers. It may include quality.

[0241] Furthermore, antibodies contain radioactive metal ions such as alpha emitters like 213Bi, and 131I Radiation including, but not limited to, n, 131LU, 131Y, 131Ho, and 131Sm. Macrocyclic chelating agents useful for conjugating metal ions into polypeptides. It can be conjugated to any therapeutic part. In certain embodiments, a large ring The chelating agent can be attached to the antibody via a linker molecule, specifically 1,4,7,10-teto It is laazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA). In this technical field, such linker molecules are generally known, and each of them is referred to as such. It is incorporated throughout the whole, 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 This is described in Med. Biol. 26(8):943~50.

[0242] Techniques for conjugating the therapeutic portion into an antibody are well known, for example, Arno n et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, M Onoclonal Antibodies and Cancer Therapy, Reisfeld et al. (eds.), pp. 243-256 (Alan R. Li ss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery,” Controlled Drug Delivery (2nd edition), Robinson et al. (eds.), pp. 623-623 (Marcel Dekker, Inc. 1987); Thorpe , “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, Monoclo nal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), 475~ 506 pages (1985); “Analysis, Results, And Future Prospective Of The Therapeutic Us” e Of Radiolabeled Antibodies In Cancer Therapy”, Monoclonal Antibodies For Cancer Detection and Therapy, Baldwin et al. (eds.), pp. 303-316 (Academic Press 1985); See Thorpe et al., 1982, Immunol. Rev. 62:119-58.

[0243] Antibodies can also adhere to solid supports, which is particularly useful for immunoassays or the purification of target antigens. It can also be done that way. Such solid supports include glass, cellulose, and polyacrylamine. This includes nylon, polystyrene, polyvinyl chloride, or polypropylene, but these are not included. Not limited.

[0244] Methods for producing antibodies nucleic acids that encode antibodies The present invention comprises a segment or domain of the FXIa-binding antibody chain described above. The present invention provides substantially purified nucleic acid molecules that encode lipeptides. The nucleotide sequence encoding the heavy chain variable region shown in SEQ ID NO: 10 or 30 , and / or nucleos encoding the light chain variable region shown in Sequence ID No. 20 or 40 It contains a cytoplasmic sequence. In specific embodiments, the nucleic acid molecule is the nucleic acid molecule identified in Table 1. The present invention includes several other nucleic acid molecules, which are identified in Table 1. Substantially identical (e.g., at least 65%, 80%, 95%, or 99%) to the Otid sequence. It contains a nucleotide sequence. When expressed from an appropriate expression vector, these polynuclei The polypeptide encoded by leotide binds to FXI and / or FXIa antigens. It is possible to demonstrate one's abilities.

[0245] In this invention, also, less than the heavy chain or light chain of the FXIa-binding antibody described above Polynucleotides that encode one CDR region, and typically all three CDR regions. Also provided. Several other polynucleotides are FXIa-binding antibodies as shown above. It encodes all or substantially all of the variable region sequence of the heavy chain and / or light chain. Due to the degenerate nature of the genome, various nucleic acid sequences encode each of the immunoglobulin amino acid sequences. It will probably happen.

[0246] The nucleic acid molecule of the present invention can encode both the variable region and the constant region of an antibody. Some of the nucleic acid sequences are substantially the same as the heavy chain sequence shown in SEQ ID NO: 11 or 31. For example, nuclei that encode identical heavy chain sequences (at least 80%, 90%, or 99%) It contains ocide. Several other nucleic acid sequences are light chain sequences shown in SEQ ID NO: 21 or 41. A light chain sequence that is substantially (for example, at least 80%, 90%, or 99%) identical to that of a 2D model. It contains nucleotides.

[0247] Polynucleotide sequences are used in de novo solid-phase DNA synthesis, or by FXIa-binding antibodies or For existing sequences encoding the binding fragment (for example, the sequences described in the examples below) It can be produced by mutation induction by PCR. Direct chemical synthesis of nucleic acids is Narang et al., 1979, Phosphotryester method by Meth. Enzymol. 68:90; Brown et al., Meth. Phosphodiester method according to Enzymol. 68:109, 1979; Beaucage et al., Tetra. Lett., 22:185 9. Diethyl phosphoramidite method according to 1981; and U.S. Patent No. 4,458,066 This can be achieved by methods known in the art, such as the solid support method described in the specification. The introduction of mutations into polynucleotide sequences by PCR is, for example, PCR Technology. In: Principles and Applications for DNA Amplification, HA Erlich (ed.), Freeman. Press, NY, NY, 1992; PCR Protocols: A Guide to Methods and Applications, Innis et al. (eds.), Academic Press, San Diego, CA, 1990; Mattila et al., Nucleic Acids Res. 19:96 7, 1991; and described in Eckert et al., PCR Methods and Applications 1:17, 1991. It can be implemented as described.

[0248] The present invention also includes the FXI-binding antibody and / or FXIa-binding antibody described above. Expression vectors and host cells for constructing the body are also provided. A variety of expression vectors are available. Using this, polynucleotides encoding FXIa-binding antibody chains or binding fragments are expressed. This can be done. Virus-based expression vectors and non-viral expression vectors Even when using a differential, antibodies can be produced within mammalian host cells. Turbo and nonviral systems express plasmids, typically proteins or RNA. Episome vectors with expression cassettes for this purpose, and human artificial chromosomes (e.g.) (See Harrington et al., Nat Genet 15:345, 1997). For example, mammals (for example) FXIa-binding polynucleotides and LOX-1-binding polypeptides in human cells Nonviral vectors useful for expression include pThioHis A, pThioHis B, and pThioHis C, pcDNA3.1 / His, pEBVHis A, pEB VHis B and pEBVHis C (Invitrogen, San Diego) CA), MPSV vectors, and for expressing other proteins, in the art. This includes many other known vectors. Useful viral vectors include retroviruses, adenoviruses. Vectors based on noviruses, adeno-associated viruses, herpesviruses, and SV40. Vectors, papillomavirus, HBP Epstein-Barr virus, vaccinia virus Includes Svector and Semlik Forest Fever Virus (SFV). Brent et al., previously cited; Smith, See Annu. Rev. Microbiol. 49:807, 1995; and Rosenfeld et al., Cell 68:143, 1992. I want to.

[0249] The selection of an expression vector depends on the intended host cell in which the vector will be expressed. The expression vector is a polynucleotide encoding an FXIa-binding antibody chain or fragment. It contains an operablely coupled promoter and other regulatory elements (e.g., enhancers). This is typical. In some embodiments, except under induction conditions, the inserted sequence An inductive promoter is used to prevent the expression of [the substance]. An inductive promoter is, for example, Arabinose, lacZ, metallothionein promoter, or heat shock promoter Includes transformers. Cultures of transformed organisms show that their expression products are better expressed in host cells. It is possible to propagate the population of acceptable code sequences under uninducible conditions without bias. In addition to the promoter, other regulatory elements also affect the FXIa-binding antibody chain. These may be required or desired for the efficient expression of fragments. These elements are A It typically includes the TG start codon and adjacent ribosome-binding sites or other sequences. In addition The efficiency of expression can be enhanced by incorporating enhancers suitable for the cell line being used. It is also possible (for example, Scharf et al., Results Probl. Cell Differ. 20:125, 1994; See also Bittner et al., Meth. Enzymol., 153:516, 1987. For example, SV40 Increase expression in mammalian host cells using an enhancer or CMV enhancer. It is possible.

[0250] The expression vector also encodes the polypeptide by the inserted FXIa-binding antibody sequence. The location of the secretory signaling sequence can also be determined to form a fusion protein with cytoplasm. The imported FXI-binding antibody sequence and / or FXIa-binding antibody sequence are contained within the vector. Often, the signal sequence is ligated before incorporation. FXI-binding antibodies and / or or sequences encoding the light chain variable domain and heavy chain variable domain of the FXIa-binding antibody The vectors used to receive may also be the constant region or part of them. It also encodes the variable region as a fusion protein with the constant region. This enables expression, which in turn leads to the creation of intact antibodies or fragments thereof. Such steady-state regions are typically human steady-state regions.

[0251] Possessing and / or expressing FXI-binding antibody chains and / or FXIa-binding antibody chains. The host cell for this can be a prokaryotic cell or a eukaryotic cell. (E. coli) ) is a prokaryotic molecule useful for cloning and expressing the polynucleotides of the present invention. It is a physical host. Other suitable microbial hosts include rod-shaped bacteria such as Bacillus subtilis. Bacteria, and species of the genera Salmonella, Serratia, and various species of bacteria This includes other Enterobacteriaceae species, such as the genus Pseudomonas. These are the prokaryotic hosts. Within it, it also typically contains expression regulatory sequences (e.g., origins of replication) that are compatible with the host cell. Expression vectors can also be created using lactose promoter systems and tryptose. Fan (trp) promoter system, beta-lactamase promoter system, or far There are also various well-known promoters, including those derived from dilambda. The promoter typically controls expression by an operator sequence through optional selection. However, this includes ribosome-binding site sequences for initiating and terminating transcription and translation. It also has. In order to express the FXIa-binding polypeptide of the present invention, other microorganisms such as yeast are used. Materials can also be used. In addition, insect cells can be combined with baculovirus vectors. Cells can also be used.

[0252] In some preferred embodiments, the FXI-binding polypeptide and / or the present invention are used. Mammalian host cells are used to express and produce FXIa-binding polypeptides. Mammalian host cells are any normal non-immortalized animal cells or normal immortalized animal cells or This includes non-normal immortalized animal cells or human cells. For example, CHO cell lines, diverse Cos cells. intact immunoglobulins, including HeLa cells, myeloma cell lines, and transformed B cells. Numerous suitable host cell lines capable of secreting brin have been developed. For the use of mammalian tissue cell cultures to express [the specified gene], see, for example, Winnacker, F. This is generally discussed in ROM GENES TO CLONES, VCH Publishers, NY, NY, 1987. Expression vectors for mammalian host cells include the origin of replication, promoter, and enhancer. See, for example, Queen et al., Immunol. Rev. 89:49~68, 1986, for expression regulatory sequences such as those of sensors. (and also ribosome-binding sites, RNA splice sites, and polyadenylation sites) , and may include necessary processing information sites, such as transcription terminator sequences.

[0253] These expression vectors are typically promoters derived from mammalian genes or mammalian genes. Contains a virus-derived promoter. A suitable promoter is a constitutive promoter. - Cell type-specific promoters, stage-specific promoters, and / or modulate It can be a controllable promoter or a modifiable promoter. Useful promoters are Tarothionein promoter, constitutive adenovirus major late promoter, dexamethasone. Zon-inducible MMTV promoter, SV40 promoter, MRP polIII promoter promoter, constitutive MPSV promoter, tetracycline-inducible CMV promoter (H (e.g., initial CMV promoters), constitutive CMV promoters, and publicly available technologies in this field. This includes, but is not limited to, combinations of knowledge promoters and enhancers.

[0254] The method for introducing an expression vector containing the target polynucleotide sequence depends on the cell host species. It varies depending on the type. For example, calcium chloride transfection is generally performed on prokaryotic cells. While it is utilized, calcium phosphate treatment or electroporation is used for other cell hosts. It is possible (see Sambrook et al., previously mentioned). Other methods include, for example, electricity. Perforation, calcium phosphate treatment, liposome-mediated transformation, injection and microinjection Cation, gene gun law, wirosom, immunoliposome, polycation: nucleic acid conjugate Gate, naked DNA, artificial virion, herpesvirus structural protein (VP) Fusion with 22 (Elliot and O'Hare, Cell 88:223, 1997), drug-enhancing uptake of DNA This includes transduction in vivo and the long-term high performance of recombinant proteins. For yield production, stable expression is often desired. For example, FXIa-binding antibodies. Cell lines that stably express chains or binding fragments are considered to be the origins of viral replication or endogenous expression. Using the expression vector of the present invention, which contains elements and a selectable marker gene, It can be prepared. After introducing the vector, the cells are kept in a fortified medium for 1-2 days. The cells can be grown and then switched to a selective medium. The purpose of the selection marker is to facilitate selection. It confers resistance to the introduced agent, and its presence enables cell growth, thereby allowing the introduced agent to... The goal is to successfully express the row in a selective medium. Stable transfection with resistance is achieved. The selected cells can be propagated using a tissue culture method appropriate for the cell type.

[0255] Framework or Fc operation The manipulated antibodies of the present invention are, for example, modified to improve the properties of the antibody, VH and / or VL. This includes a modified antibody in which the framework residues within the antibody have been modified. Typically, the treatment involves applying a substance to reduce the immunogenicity of the antibody. For example, one technique... This "abruptly returns" one or more framework residues to the corresponding germline sequence. It is about "causing mutation." More specifically, antibodies that have undergone somatic mutation are derived from antibodies It may contain framework residues that differ from the germline sequence. Such residues may be antibodies The framework sequence is identified by comparing it with the germline sequence from which the antibody originates. This is possible. To return the framework region sequence to the three-dimensional arrangement of those germline cells. For example, site-directed mutagenesis can induce somatic mutations in germline sequences. It is possible to "induce reverse mutations." Such "reverse-mutated" antibodies are also This invention is intended to encompass the following.

[0256] Another type of framework modification involves residues within one or more framework regions. Alternatively, by mutating one or more residues within the CDR region, T cell epithelial cells can be mutated. This involves removing the p-component, thereby reducing the potential immunogenicity of the antibody. It is also referred to as "deimmunization," as described in U.S. Patent Publication No. 20030153043 by Carr et al. Further details are provided in the detailed specifications.

[0257] In addition to, or alternatively to, any modifications made within the framework or CDR area The antibodies of the present invention include modifications within the Fc region, typically resulting in a longer serum half-life, improved complement binding, and increased F One or more antibodies, such as binding to the C receptor and / or antigen-dependent cell-mediated cytotoxicity. It is also possible to manipulate the functional properties of the antibody to be modified. Furthermore, the antibody of the present invention is chemical It can also be modified (for example, one or more chemical parts can be attached to the antibody). (to be able to), and by modifying its glycosylation, here again, one or more functional characteristics of the antibody It is also possible to modify the gender. Each of these embodiments is described below. Further details are provided below. The numbering of residues within the Fc region is done using the EU index by Kabat. It's a kus.

[0258] In one embodiment, the number of cysteine ​​residues in the hinge region is modified, for example, by increasing or Modify the hinge region of CH1 to reduce it. This technique was developed by Bodmer et al. Further details are provided in U.S. Patent No. 5,677,425. CH1 hinge The number of cysteine ​​residues within a region, for example, facilitates the assembly of the light and heavy chains. or modify the antibody to increase or decrease its stability.

[0259] In another embodiment, the Fc hinge region of the antibody is mutated to alter the biological half-life of the antibody. To shorten. More specifically, the binding of the antibody to Staphylococcus protein A (SpA) is In contrast to the binding of the Fc-hinge domain to SpA, one or more Amino acid mutations are introduced into the interface region between the CH2 domain and CH3 domain of the Fc-hinge fragment. This method is introduced in U.S. Patent No. 6,165,745 by Ward et al. Further details are provided there.

[0260] In another embodiment, the antibody is modified to extend its biological half-life. This is possible. For example, in U.S. Patent No. 6,277,375 by Ward, One or more of the mutations described may be used. Alternatively, To extend the biological half-life, antibodies were used in a study by Presta et al. in the U.S., which found 5,869 Specification No. 046; and Specification No. 6,121,022, as described in I Salvage receptor binding derived from two loops of the CH2 domain in the Fc region of γ The CH1 region or CL region can also be modified to contain an epitope.

[0261] In yet another embodiment, at least one amino acid residue is used to enhance the effector function of the antibody. The Fc region is modified by replacing it with a different amino acid residue. For example, The antibody has modified its affinity for the effector ligand, but the parent antibody's antigen... To maintain binding ability, one or more amino acids are replaced with different amino acid residues. It is possible. An effector ligand that modifies the affinity for it is, for example It could be an Fc receptor or the C1 component of complement. Both methods were developed by Winter et al. U.S. Patent No. 5,624,821 and U.S. Patent No. 5,648,260 Further details are provided in [the document].

[0262] In another embodiment, the antibody modifies binding to C1q and / or causes complement-dependent cell damage. To reduce or eliminate the harmful effects (CDC), one amino acid residue is selected. Alternatively, multiple amino acids can be replaced with different amino acid residues. This method is I Further details are provided in U.S. Patent No. 6,194,551 by Dusogie et al. It is listed.

[0263] In another embodiment, one or more amino acid residues are modified to thereby bind to complement. This method modifies the ability of the antibody to bind. This technique was developed by Bodmer et al. in the PCT Public Published International Publication No. Further details are provided in pamphlet number 94 / 29351.

[0264] In specific embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing V1401 VL CDR and VH CDR are used in any surface-associated FXI To reduce the likelihood of ADCC or CDC being caused, two amino acids Human IgG (e.g., IgG1) Fc region containing substitutions D265A and P329A. This includes the region. These alanine substitutions have been shown to reduce ADCC and CDC. (For example, Idosugie et al., J. Immunol. 164:4178-4184, 2000; Shields et al., J. See Biol. Chem. 276:6591-6604, 2001.

[0265] In yet another embodiment, by modifying one or more amino acids, antibody-dependent To increase the ability of antibodies that mediate cytotoxicity (ADCC) and / or Fcγ receptor The Fc region is modified to increase the antibody's affinity for the substance. This is stated in Presta's PCT Public International Publication No. 00 / 42072. Furthermore, it is stated that FcγRI, FcγRII, and Fcγ on human IgG1 are also mentioned. Binding sites for RIII and FcRn are also mapped, and improved binding has been observed. Other forms have also been described (see Shields, RL et al., 2001 J. Biol. Chen. 276:6591~6604). I want to be treated that way.

[0266] In yet another embodiment, the glycosylation of the antibody is modified. For example, deglycosylated antibody It is possible to create a body (i.e., an antibody lacking glycosylation). Glycosylation is For example, it is possible to modify the antibody to increase its affinity for the "antigen". Such carbohydrate modifications include, for example, one or more glycosylations within an antibody sequence. This can be achieved by modifying the parts. For example, one or more variable regions One or more amino acids that result in the loss of the framework glycosylation site. By creating a substitution, glycosylation at that site can be eliminated. This deglycosylation process increases the affinity of antibodies against the antigen. This can be done. Such a method is described in U.S. Patent No. 5,714,350 by Co et al.; This is further described in Patent No. 6,350,861.

[0267] In addition, or alternatively, a low-fucosylated antibody or bifidobacterial antibody with a reduced amount of fucosyl residues. We will produce antibodies with modified glycosylation types, such as antibodies with increased GlcNac structure. This modification of the glycosylation pattern can increase the ADCC activity of the antibody. This has been supported. Such carbohydrate modifications, for example, affect the glycosylation mechanism. This can be achieved by expressing antibodies in modified host cells. The text describes cells with a modified glycosylation mechanism, and describes how to express the recombinant antibody of the present invention. This allows us to use them as host cells for producing antibodies with modified glycosylation. This is possible. For example, EP1,176,195 by Hang et al. describes how to do this within a cell line. The antibody expressed in this manner exhibits low fucosylation, and fucosyltransferase is used to This document describes cell lines in which the FUT8 gene, which controls FUT8, is functionally disrupted. The PCT Public International Publication No. 03 / 035835 by Asn( 297) Reduces the ability to attach to linked carbohydrates, and also reduces the expression within the host cell. Lecl, a mutant CHO cell line, also results in the low fucosylation of the antibodies. The text describes three cell types (see also Shields, RL et al., 2002 J. Biol. Chem. 277:26733). See also ~26740). PCT Public Publication No. 99 / 54342 by Umana et al. The pamphlet states that antibodies expressed in a manipulated cell system resulted in increased ADCC activity of the antibodies. Modify the glycoprotein to exhibit an increase in the bifurcated GlcNac structure. Glycosyltransferases (e.g., beta(1,4)-N-acetylglucosamine) Cell lines engineered to express niltransferase III (GnTIII) This is described (see also Umana et al., 1999 Nat. Biotech. 17:176~180). stomach).

[0268] Methods for manipulating modified antibodies As discussed above, the VH sequence and VL sequence or full-length heavy chain and FXIa-binding antibodies that have a full-length light chain sequence contain a full-length heavy chain sequence and / or a full-length light chain sequence. Modify the column, VH sequence and / or VL sequence, or the constant region attached thereto. This can be used to create new FXIa-binding antibodies. In another aspect of the present invention, the structural features of the FXIa-binding antibody of the present invention are used to... To bind to FXIa and also inhibit one or more functional properties of FXIa (for example) This inhibits the binding of FXIa to the FXIa receptor and inhibits FXIa-dependent cell proliferation. ) and other structurally related FX that retain at least one functional property of the antibody of the present invention. To create an Ia-binding antibody.

[0269] For example, one or more CDR regions of the antibody of the present invention or mutations thereof are recombinant Therefore, in combination with known framework areas and / or other CDRs, recombinant The present invention, as described above, manipulates the process to create further FXIa-binding antibodies. This is possible. Other types of modifications include those described in the previous section. Starting materials for the operation. The material is one or more of the VH and / or VL sequences presented herein. This refers to a number of CDR regions, or one or more of them. One or more of the VH sequences and / or VL sequences presented in the specification, This involves actually preparing antibodies that have one or more of those CDR regions (i.e., It is not necessary to express it as an protein. Rather, it is contained within the sequence. The information is used as starting material to create a "second generation" sequence derived from the original sequence. Next, the "second generation" sequence is prepared as a protein and expressed.

[0270] Therefore, in another embodiment, the present invention is selected from the group consisting of Sequence IDs 3 and 23. The CDR1 sequence, the CDR2 sequence selected from the group consisting of sequence numbers 4 and 24, Rabini / or a heavy CDR3 sequence selected from the group consisting of sequence numbers 5 and 25 A CDR1 sequence selected from the group consisting of the antibody sequence of the chain variable region and SEQ ID NOs: 13 and 33. A CDR2 sequence selected from the group consisting of columns, sequence numbers 14 and 34, and / or Light chain variable region having a CDR3 sequence selected from the group consisting of sequence numbers 15 and 35 Prepare an FXIa-binding antibody consisting of an antibody sequence and / or the antibody sequence of the heavy chain variable region and / or This involves modifying at least one amino acid residue in the antibody sequence of the light chain variable region, at least A method for creating a modified antibody sequence and expressing the modified antibody sequence as a protein. To provide.

[0271] Therefore, in another embodiment, the present invention selects from the group consisting of Sequence IDs 6 and 26. The CDR1 sequence, the CDR2 sequence selected from the group consisting of sequence numbers 7 and 27, Rabini / or a heavy CDR3 sequence selected from the group consisting of sequence numbers 8 and 28 A CDR1 sequence selected from the group consisting of the antibody sequence of the chain variable region and SEQ ID NOs: 16 and 36. A CDR2 sequence selected from the group consisting of columns, sequence numbers 17 and 37, and / or Light chain variable region having a CDR3 sequence selected from the group consisting of sequence numbers 18 and 38 Prepare an FXIa-binding antibody consisting of an antibody sequence and / or the antibody sequence of the heavy chain variable region and / or This involves modifying at least one amino acid residue in the antibody sequence of the light chain variable region, at least A method for creating a modified antibody sequence and expressing the modified antibody sequence as a protein. To provide.

[0272] Therefore, in another embodiment, the present invention optimizes expression in mammalian cells. An FXIa-binding antibody, wherein the sequence is selected from the group of sequence numbers 11 or 31. The full-length heavy chain antibody sequence and the sequence selected from the group of SEQ ID NOs. 21 or 41 A long light chain antibody sequence and an FXIa-binding antibody are prepared, and the full-length heavy chain antibody sequence and / or This involves modifying at least one amino acid residue in the full-length light chain antibody sequence, thereby modifying at least one modified This invention provides a method for creating modified antibody sequences and expressing these modified antibody sequences as proteins. In one embodiment, the modification of the heavy or light chain is performed within the framework region of the heavy or light chain. That is the case.

[0273] The modified antibody sequence is also described in the CDR3 sequence, or US2005 / 0255552. The CDR1 and CDR2 sequences are listed, with the minimum essential binding determinants fixed. This can also be prepared by screening a diverse antibody library. This can be done. Screening involves using phage display technology, etc., to test antibodies in an antibody library. It is possible to carry out screening according to any appropriate screening technique for screening from Lee. can.

[0274] Modified antibody sequences can be prepared and expressed using standard molecular biology techniques. The antibodies encoded by the modified antibody sequence are human, cynomolgus monkey, rat, and / or The antibody specifically binds to mouse FXIa; and the antibody is used in F36E cell proliferation assays. and / or in the Ba / F3-FXIaR cell proliferation assay, FXIa-dependent cell proliferation This specification includes, but is not limited to, inhibiting cytoplasmic proliferation of FXIa cells. This antibody retains one, some, or all of the functional characteristics of a composite antibody.

[0275] In certain embodiments of the method for manipulating the antibody of the present invention, the FXIa-binding antibody coding sequence It is possible to introduce mutations randomly or selectively along all or part of the gene. The resulting modified FXIa-binding antibody is subjected to the binding activity described herein. It can be screened for both and / or other functional characteristics. In this technical field, Mutation methods are described. For example, Short's PCT Public Publication No. 02 / 0 Pamphlet No. 92780 covers saturated mutagenesis, synthetic ligation assembly, and more. Alternatively, these combinations can be used to create antibody mutations and screen for them. This document describes the law. Alternatively, see the PCT International Publication No. 03 / 0 by Lazar et al. Pamphlet No. 74679 describes how to use computer-based screening methods to detect antibodies. This document describes methods for optimizing physiological and chemical properties.

[0276] In certain embodiments of the present invention, the antibody is manipulated to remove the deamidation site. Deamidation causes structural and functional changes within peptides or proteins. It is known that this can occur. Deamide deamide can lead to a decrease in biological activity, as well as in protein drugs. This can also result in alterations to the pharmacokinetics and antigenicity of the drug (Anal Chem. March 1, 2005; 77). (5):1432~9).

[0277] In certain embodiments of the present invention, antibodies increase pI and improve their drug-like properties. It is manipulated to do so. The pI of a protein is key to the overall biophysical properties of the molecule. This is a determining factor. Antibodies with a small pI have low solubility, low stability, and are prone to aggregation. It is well known that... Furthermore, the purification of antibodies with low pI is particularly useful in clinical applications. Scaling up for use can be difficult and problematic. By increasing the pI of anti-FXI antibody and / or anti-FXIa antibody or Fab This improves their solubility and allows for the formulation of antibodies at high concentrations (>100 mg / ml). This became possible by formulating antibodies at high concentrations (e.g., >100 mg / ml). This brings the advantage of being able to administer high doses of antibodies, which in turn allows for the treatment of blood clots. It offers significant advantages for treating chronic diseases, including sexual disorders and / or thromboembolic disorders. This can enable a reduction in the frequency of administration. Increasing pI also affects the IgG version of the antibody. This also increases the recycling of FcRn, which allows the drug to be used for longer periods of time. It can remain in the body and potentially reduce the number of injections required. Finally, pI Due to the increase in [specific factor], the overall stability of the antibody is significantly improved, and as a result, the storage life is extended. This also leads to an extension of biological activity in vivo. pI is 8.2 or higher. It is preferable that this be the case.

[0278] The functional properties of the modified antibody are determined by assays (e.g., ELISA) shown in the examples. And, standard A available in the art and / or described herein It can be evaluated using the ssey.

[0279] Preventive and therapeutic use Antibodies that bind to FXI and / or FXIa as described herein (e.g., NO Table 1 shows examples of anti-FXI / FXIa antibodies, including V1401 VL CDR and VH CDR. The antibody described in [the document] contains an effective amount of the antibody or antigen-binding fragment of the present invention, which is necessary to By administering to the target population, thromboembolic diseases or thromboembolic disorders (for example, Thrombotic stroke, atrial fibrillation, prevention of stroke in atrial fibrillation (SPAF), deep vein thrombosis , venous thromboembolism, pulmonary embolism, acute coronary syndrome (ACS), ischemic stroke, acute lower limb collapse To treat (hemorrhagic pulmonary hypertension, chronic thromboembolic pulmonary hypertension, or systemic embolism), therapeutically It can be used at various concentrations. The present invention provides an effective amount of the antibody of the present invention, which can be used as needed. By administering it to the target, it is used to treat thromboembolic disorders (e.g., thrombotic disorders). The present invention provides a method for administering an effective amount of the antibody of the present invention to a subject that requires it. This can lead to thromboembolic disorders (for example, thrombotic stroke, atrial fibrillation, and brain damage in atrial fibrillation). Prevention of stroke (SPAF), deep vein thrombosis, venous thromboembolism, pulmonary embolism, acute coronary artery syndrome Acute thromboembolic stroke (ACS), ischemic stroke, acute lower limb ischemia, chronic thromboembolic pulmonary hypertension, or systemic thromboembolic stroke. This provides a method for treating uterine embolism.

[0280] The antibodies described herein (e.g., NOV1401, or VL of NOV1401) Anti-FXI / FXIa antibodies, including CDR and VH CDR, as listed in Table 1. The body includes, in particular, but is not limited to, thrombotic disorders, as described in more detail herein. Used to treat, prevent, and improve undefined thromboembolic conditions or thromboembolic disorders. It can be used.

[0281] The antibodies presented herein (e.g., VL CDR and VH CD of NOV1401) Antibodies (such as anti-FXI / FXIa antibodies containing R, as listed in Table 1) are also thromboembolic. It can also be used in combination with other drugs to prevent, treat, or improve disorders. For example, statin therapy is used to treat patients with thrombotic and / or thromboembolic disorders. To use in combination with the FXIa antibody and antigen-binding fragment of the present invention, It is possible.

[0282] In specific embodiments of this specification, the treatment or prevention of stroke in patients with atrial fibrillation A method for stopping the process, comprising an effective amount of the anti-FXI / FXIa antibody described herein, for example. Anti-F containing NOV1401, or VL CDR and VH CDR of NOV1401 Anti-FXI / FXIa antibodies, such as XI / FXIa antibodies, as listed in Table 1, as needed. A method is presented that includes the step of administering the drug to a patient.

[0283] In specific embodiments of this specification, embolic stroke and total stroke in patients with atrial fibrillation Individuals who manage or prevent risks or conditions associated with atrial fibrillation (AF), such as physical embolism. A method comprising an effective amount of the anti-FXI / FXIa antibody described herein, for example, NOV Anti-FXI / F, including VL CDRs and VH CDRs of 1401, or NOV1401. Anti-FXI / FXIa antibodies, such as XIa antibodies, as listed in Table 1, are available to patients who require them. A method is presented that includes the step of administering the substance to the person.

[0284] In specific embodiments of this specification, embolic stroke and total stroke in patients with atrial fibrillation Methods for treating, managing, or preventing conditions associated with atrial fibrillation (AF), such as physical embolism. There is an effective amount of the anti-FXI / FXIa antibody described herein, for example, NOV14 Anti-FXI / FXI including VL CDRs and VH CDRs of 01, or NOV1401 Anti-FXI / FXIa antibodies, such as antibody α, as listed in Table 1, should be administered to patients who require them. A method is presented which includes the step of administering the following. In a particular embodiment, an AF patient has bleeding It is highly dangerous.

[0285] In specific embodiments of this specification, the subject (for example, those with deep vein thrombosis or...) In individuals at risk of developing this condition, treatment for deep vein thrombosis or related conditions is administered. Methods for placing, controlling, or preventing an effective amount of anti-FXI / F as described herein. XIa antibodies, e.g., NOV1401, or VL CDR and VH of NOV1401 Anti-FXI / FXIa antibodies, including CDRs, as listed in Table 1. A method is presented that includes the step of administering an antibody to a target that needs it.

[0286] In specific embodiments of this specification, the subject (for example, with or with venous thromboembolism) is referred to as Venous thromboembolism (VTE) or related conditions in individuals at risk of developing this condition. A method for treating, controlling or preventing a condition, comprising an effective amount of the antimicrobial agent described herein. FXI / FXIa antibody, e.g., NOV1401, or VL CDR of NOV1401 And anti-FXI / FXIa antibodies including VH CDR, as listed in Table 1. A method is presented that includes the step of administering the / FXIa antibody to a target that requires it. In certain embodiments, subjects treated with the anti-FXI / FXIa antibodies presented herein This includes cancer patients, and includes 1) first-time idiopathic VTE with a low risk of bleeding, and 2) idiopathic VTE. The patient has experienced a recurrence of the disease, or 3) VTE associated with a thrombotic diathesis.

[0287] In specific embodiments of this specification, the subject (for example, a patient with or developing pulmonary embolism) is considered. In individuals at risk of developing the disease, treatment, management, and treatment of pulmonary embolism or related conditions. or a method of prevention, comprising an effective amount of the anti-FXI / FXIa antibody described herein. For example, including NOV1401, or VL and VH CDRs of NOV1401. Anti-FXI / FXIa antibodies, such as those listed in Table 1, A method is presented that includes the step of administering the substance to a target that requires it.

[0288] In specific embodiments of this specification, acute coronary syndrome (ACS) or the subject is described. A method for treating, managing, or preventing a condition related thereto, in an effective amount, as specified herein. The anti-FXI / FXIa antibody listed, for example, NOV1401, or NOV1401's V Table 1 lists anti-FXI / FXIa antibodies, including L CDR and VH CDR. A method comprising the step of administering an anti-FXI / FXIa antibody to a subject in need thereof. It will be presented.

[0289] In specific embodiments of this specification, the subject (for example, those with or suffering from ischemic stroke) is referred to. Methods for treating, managing, or preventing ischemic stroke in individuals at risk of developing ischemic stroke. and an effective amount of the anti-FXI / FXIa antibody described herein, for example, NOV1 Anti-FXI / FX, including VL and VH CDRs of 401 or NOV1401. Anti-FXI / FXIa antibodies, such as Ia antibodies, as listed in Table 1, should be used for the target population that requires them. A method is presented that includes the step of administering to [the substance].

[0290] In specific embodiments of this specification, acute lower limb ischemia in a subject is treated, managed, or A method of prevention, comprising an effective amount of the anti-FXI / FXIa antibody described herein, for example If, then, anti- FXI / FXIa antibodies, etc., as listed in Table 1, must be used. A method is presented that includes the step of administering the drug to the target subject.

[0291] In specific embodiments of this specification, chronic thromboembolic pulmonary hypertension in a subject is treated and managed A method for treating or preventing the use of an effective amount of anti-FXI / FXIa as described herein. Antibodies, e.g., NOV1401, or VL CDR and VH CD of NOV1401 Anti-FXI / FXIa antibodies, including those containing R, as listed in Table 1. A method is presented that includes the step of administering it to a person who needs it.

[0292] In specific embodiments of this specification, the subject (for example, with or with systemic embolism) is a subject (e.g., with systemic embolism or similar). Methods for treating, managing, or preventing systemic embolism in individuals at risk of developing the condition. and an effective amount of the anti-FXI / FXIa antibody described herein, for example, NOV1 Anti-FXI / FX, including VL and VH CDRs of 401 or NOV1401. Anti-FXI / FXIa antibodies, such as Ia antibodies, as listed in Table 1, should be used for the target population that requires them. A method is presented that includes the step of administering to [the substance].

[0293] In certain embodiments of this specification, a catheter becomes thrombosed (for example, In cancer patients, this is a Hickman catheter-related condition, or the tubing is clotting. Treating, managing, or preventing thromboembolic states caused by extracorporeal membrane oxygenation (ECMO). A method for stopping the process, comprising an effective amount of the anti-FXI / FXIa antibody described herein, for example. Anti-F containing NOV1401, or VL CDR and VH CDR of NOV1401 Anti-FXI / FXIa antibodies, such as XI / FXIa antibodies, as listed in Table 1, as needed. A method is presented that includes the step of administering the drug to the target subject.

[0294] In certain embodiments, an anti-FXI / FXIa antibody as described herein, for example, NOV Anti-FXI / F, including VL CDRs and VH CDRs of 1401, or NOV1401. Treatment with anti-FXI / FXIa antibodies, such as XIa antibodies, as listed in Table 1 is required. The target is, • Indications for chronic anticoagulation therapy (e.g., AF, left ventricular thrombosis, history of cardioembolic stroke) Target; • Patients at moderate to high risk of massive bleeding; • Dual antiplatelet therapy (aspirin and P2Y1) to prevent stent thrombosis. Elective percutaneous coronary intervention may require stent placement (administering a receptor antagonist). Patients undergoing percutaneous transluminal intervention (PCI) or direct PCI It may include.

[0295] In a particular embodiment, the following state: Paroxysmal atrial fibrillation or paroxysmal atrial flutter, persistent atrial fibrillation or persistent atrial flutter, Alternatively, if cardiac arrhythmias such as persistent atrial fibrillation or persistent atrial flutter are suspected or confirmed. Thromboembolism in the subject being treated; • Subjects with atrial fibrillation requiring stroke prevention (SPAF), and whose subgroups are The target group consists of AF patients undergoing percutaneous coronary intervention (PCI); • Management of acute venous thromboembolic events (VTE) in patients at high risk of bleeding. and prevention of long-term secondary VTE; • In secondary prevention following transient ischemic attack (TIA) or non-functional stroke In the prevention of cerebral and cardiovascular events, as well as thromboembolic events in heart failure with sinus rhythm. Cerebral and cardiovascular events; • Clot formation and thromboembolism in the left atrium in patients undergoing cardiac defibrillation for cardiac arrhythmias. Embolism; Thrombosis before, during, and after ablation procedures for cardiac arrhythmias; • Venous thrombosis, which is deep vein thrombosis or superficial vein thrombosis in the lower or upper extremities. Vessel thrombosis, thrombosis in the abdominal and thoracic veins, sinus thrombosis and jugular vein thrombosis This includes, but is not limited to, treatment and secondary prevention; Thrombosis on any artificial surface within a vein, such as a catheter or pacemaker lead; • Pulmonary embolism in patients with or without venous thrombosis; Chronic thromboembolic pulmonary hypertension (CTEPH); • Arterial thrombosis on ruptured atherosclerotic plaque, on arterial prostheses or catheters These are thrombosis in the arteries and thrombosis in seemingly normal arteries, and these are acute coronary syndrome. Group, ST-elevation myocardial infarction, non-ST-elevation myocardial infarction, unstable angina, stent thrombosis, arterial Thrombosis of any artificial surface within the system, and in subjects with or without pulmonary hypertension. This includes, but is not limited to, pulmonary thrombosis; • Thrombosis and thrombosis in patients undergoing percutaneous coronary intervention (PCI) Embolism; • Cardioembolic stroke and cryptogenic stroke; Thrombosis in patients with invasive and non-invasive cancerous malignancies; • Thrombosis involving indwelling catheters; • Thrombosis and thromboembolism in critically ill patients; • Cardiac thrombosis and thromboembolism, including cardiac thrombosis after myocardial infarction and cardiovascular thrombosis. Conditions such as aneurysms, cardiomyopathy, cardiac hypertrophy and cardiac dysfunction, myocarditis, and artificial surfaces within the heart This includes, but is not limited to, cardiothrombosis associated with the condition; Thromboembolism in patients with valvular heart disease, with or without atrial fibrillation; • Thromboembolism involving mechanical or biological prostheses for heart valves; • After cardiac repair for simple or complex cardiac malformations, natural or artificial cardiac pads Injuries or trauma in patients with arterial or venous conduits; • Total knee replacement, total hip replacement, and orthopedic surgery, thoracic surgery, or abdominal surgery Venous thrombosis and thromboembolism following; • Following neurosurgical procedures including intracranial and spinal interventions Arterial thrombosis or venous thrombosis; • Factor V Leiden, prothrombin mutation, antithrombin III, prote Protein C deficiency and protein S deficiency, factor XIII mutations, familial fibrosis Linogenemia, congenital plasminogen deficiency, elevated factor XI levels, sickle cell disease, Antiphospholipid syndrome, autoimmune diseases, chronic bowel disease, nephrotic syndrome, hemolytic uremic disease, bone marrow Proliferative disorders, disseminated intravascular coagulation, paroxysmal nocturnal hemoglobinuria, and heparin-induced hematologic dysplasia Congenital or acquired thrombotic predisposition, including but not limited to thrombotic plaque; • Thrombosis and thromboembolism in chronic kidney disease; and • Thrombosis and Thromboembolism One of these is an anti-FXI / FXIa antibody as described herein, for example, NOV140 1, or anti-FXI / FXIa, including VL CDR and VH CDR of NOV1401. Treatment or management can be performed with anti-FXI / FXIa antibodies, such as those listed in Table 1. Cut.

[0296] In specific embodiments of this specification, anti-FXI / FXIa antibodies presented herein (e.g., , anti-FXI / FXIa antibodies including VL CDR and VH CDR of NOV1401, etc. Bleeding in patients treated with or administered with antibodies (listed in Table 1) For example, a method for managing bleeding associated with trauma, surgery, menstruation, or postpartum, and anticoagulant Methods including reversal of solidification effects are presented. FXI deficiency is associated with spontaneous bleeding symptoms. It is rare, and in specific cases, bleeding is associated with trauma, surgery, menstruation, or postpartum. This is the most typical example. Prolonged bleeding occurs after major trauma, or in the oral mucosa, nasal mucosa, It can occur after surgery involving organs with areas of high fibrinolysis, such as the genital mucosa or urinary tract mucosa. Tooth extraction, tonsillectomy, and uterine or prostate ablation carry a high risk of bleeding. This is an example of surgery involving sexuality. Also, people with disabilities tend to develop nosebleeds and ecchymosis. The symptoms are severe, and although rare, they can also cause bleeding in the urine or intestines. FXI deficiency In patients with this condition, the frequency of spontaneous muscle or joint bleeding and intracranial hemorrhage is not increased. Pulse puncture is not typically associated with prolonged bleeding. Other gene mutations associated with FXI deficiency. This may contribute to heterogeneous and unexpected bleeding tendencies in patients with severe FXI deficiency. The combined use of platelet agents, other anticoagulants, and fibrinolytic agents may increase the risk of bleeding.

[0297] In certain embodiments of this specification, the anti-FXI / FXIa antibodies presented herein (for example) For example, an anti-FXI / FXIa antibody containing VL CDR and VH CDR of NOV1401 A method for managing bleeding in patients treated with antibodies (listed in Table 1), A method has been proposed that includes reversing the primary anticoagulant effect for a sufficient amount of time to manage the bleeding. In a specific embodiment, the step of reversing the anticoagulant effect is (i) colloid, crystal Replacement of body fluids using human plasma or plasma proteins such as albumin; (ii) a transfusion of packed red blood cells or whole blood. In certain embodiments, for example, In severe emergency cases, the therapeutic agent for reversing the effect of anticoagulants is fresh frozen plasma (F FP), prothrombin complex concentrate (PCC), and activated PCC (APCC); For example, factor VIII inhibitor bypass activity (FEIBA) and recombinant activity It includes, but is not limited to, blood components that promote hemostasis, such as factor VII (rFVIIa). It is not possible. In one embodiment, following the administration of rFVIIa at a dose of 30 μg / kg, 6 In addition to 1g of tranexamic acid every hour for 5-7 days, take 15g every 2-4 hours. A regimen including administration of rFVIIa at a dose of ~30 μg / kg over 24-48 hours. This refers to the anti-FXI / FXIa antibodies presented herein (e.g., NOV) used in major surgery. Treated with antibodies containing VL CDR and VH CDR of 1401 or NOV1401 To restore hemostasis in patients with bleeding inaccessible areas and inaccessible sites of ongoing bleeding, It may have the potential to stop bleeding. For example, Riddelle et al. showed that inhibitors are not present. We reported on our experience with four patients undergoing surgery who had severe FXI deficiency. Riddell et al., 2011, Thromb. Haemost.; 106: 521-527), the patient was given 30 μg / kg rFVIIa and 1 g of tranexamic acid were administered intravenously during induction of anesthesia. A bolus administration of 15-30 μg / kg of rFVIIa is considered rotational therapy. As indicated by the guidelines shown by the romboelastometry (ROTEM) results, 2~ It was administered at 4-hour intervals. Patients received the above-mentioned dose of rFVIIa over a period of 24-48 hours. The treatment was administered. 1g of tranexamic acid every 6 hours was continued for 5 days. The low-dose series uses a low dose of 15-30 μg / kg in combination with tranexamic acid. rFVIIa was found to be effective in correcting hemostatic abnormalities in severe FXI deficiency in this study. It was completely effective. Inhibitors (usually transfused to patients with severe FXI deficiency) This is a severe FXI deficiency accompanied by autologous FXI neutralizing antibodies acquired after administration of blood products. In another study, which included four patients who underwent five surgeries, the authors (Livnat et al., 2009, T) hromb. Haemost.; 102: 487-492) applied the following protocol: 2 hours before surgery In this case, 1 g of oral tranexamic acid, then immediately before the intervention, the patient is given a separate dose. IV tranexamic acid 1g was administered. Recombination therapy was performed with a dose in the range of 15-30 μg / kg. FVIIa was injected at the completion of the surgery. Subsequently, 1 g of oral tranexamic acid was administered over 6 hours. Each application was performed for at least 7 days. Fibrin adhesive was applied to the excisions in one patient. It was sprayed onto the extracted gallbladder bed. This protocol is for severe FXI deficiency with inhibitors. We ensured normal hemostasis in patients with the condition.

[0298] In one embodiment, fibrin adhesive is used in dental treatment for patients with FXI deficiency. It can restore local hemostasis during surgery (Bolton-Maggs (2000) Haemophilia; 6 (S1): 100-9). With the anti-FXI / FXIa antibody presented herein (e.g., NOV1401) In certain embodiments of methods for managing bleeding in a patient being treated, fibrillation In association with the use of adhesives, tranexamic acid 1g every 6 hours for 5-7 days is used. The regimen includes patients undergoing minor surgery and those experiencing bleeding events in the oral and nasal cavities. It is used to establish local hemostasis in subjects where the bleeding site is accessible. ru.

[0299] Pharmaceutical composition This invention relates to an FXIa-conjugated antibody formulated in combination with a pharmaceutically acceptable carrier (inter The present invention provides a pharmaceutical composition containing (a cerebrospinal or binding fragment). The composition also contains, for example, a thrombus. One or more other treatments suitable for the treatment or prevention of embolic disorders (e.g., thrombotic disorders) It may also contain agents. Pharmaceutically acceptable carriers enhance or stabilize the composition. Alternatively, it can be used to facilitate the preparation of compositions. pharmaceutically acceptable. The carrier is a physiologically compatible solvent, dispersion medium, coating, antimicrobial and antifungal agent, isotonic. This includes agents and absorption retarders.

[0300] The pharmaceutical composition of the present invention can be administered by various methods known in the art. The route and / or method of administration will vary depending on the desired outcome. Administration is by intravenous injection. Administration (IV), intramuscular administration (IM), intraperitoneal administration (IP), or subcutaneous administration It is preferable to administer it (sc) or in close proximity to the target site. Pharmacologically permissible The carriers can be administered intravenously, intramuscularly, subcutaneously, parenterally, spinally, or epidermally. It shall be suitable for administration (e.g., by injection or infusion). Depending on the route of administration, activation Antibodies, which are compounds, i.e., dispecific and polyspecific molecules, can either eliminate a compound or eliminate a compound. It can be coated with a material that protects against the action of potentially activating acids and other natural conditions. Cut.

[0301] In certain embodiments, the anti-FXI / FXIa antibody described herein (e.g., NOV14) Antibodies containing LCDR and HCDR of 01, or NOV1401, as listed in Table 1. The antibody (containing) is placed in a fluid vial for subcutaneous injection, with approximately 75 mg per 1 mL to 1 mL equivalent. Formulated at a concentration of approximately 200 mg. In certain embodiments, the pharmaceutical composition is carried by a pharmaceutical carrier. or pharmaceutical excipients, for example, sucrose and polysorbate 20. Specific embodiments The pharmaceutical composition contains L-histidine and / or histidine HCl monohydrate. In certain embodiments, the pharmaceutical composition has a pH of about 4-7 or 5-6.

[0302] In certain embodiments, the anti-FXI / FXIa antibody described herein (e.g., NOV14) Antibodies containing LCDR and HCDR of 01, or NOV1401, as listed in Table 1. The antibody (which is present in the body) is prepared in a fluid vial for subcutaneous injection at a concentration of 150 mg per 1 mL. Formulation is performed. In one embodiment, a 150 mg / mL fluid formulation is prepared with a pH of 5.5 ± 0.5. The medication contains 150 mg of anti-FXI / FXIa antibody, L-histidine, and histidine HCl monohydrate. It contains sucrose and polysorbate 20. The composition is sterile and fluid. Assume it is a body. Appropriate fluidity can be achieved by using a coating such as lecithin. This allows for maintenance, and in the case of a dispersion, it maintains the required particle size and the boundary This can be maintained by using surfactants. In many cases, isotonic agents are included in the composition. For example, sugars, polyhydric alcohols such as mannitol or sorbitol, and sodium chloride It is preferable to include um. Long-term absorption of the injectable composition is possible if the composition contains substances that delay absorption. By incorporating a drug such as aluminum monostearate or gelatin It can bring about.

[0303] The pharmaceutical composition of the present invention is well known in the art and is based on methods commonly used in practice. It can be prepared according to the following. For example, Remington: The Science and Practice of Pharm acy, Mack Publishing Co., 20th edition, 2000; and Sustained and Controlled Release Dru See g Delivery Systems, edited by JR Robinson, Marcel Dekker, Inc., New York, 1978. It is preferable that the pharmaceutical composition be manufactured under GMP conditions. Therefore, we will use a therapeutically effective dose or therapeutically effective dose of the FXIa-binding antibody. This is typical. FXIa-binding antibodies are obtained by conventional methods known to those skilled in the art, and are pharmaceutically effective. Formulate into an acceptable dosage form. The administration regimen is designed to achieve the desired optimal response (e.g., treatment). Adjust to produce a suitable response. For example, a single bolus may be administered, or multiple boluses may be used. The dosage can also be divided into several doses and administered over a period of time, depending on the requirements of the treatment situation. Depending on the indication, the dose may also be proportionally reduced or increased. For ease of use and uniformity of dosage, parenteral compositions are formulated in dose-unit form. This is particularly advantageous. The dosage unit forms used herein refer to the target being treated. This refers to a physically individual unit suitable as a unit dose for a particular drug. Each unit is a required drug. It contains a predetermined amount of active compound, calculated to produce the desired therapeutic effect in relation to the carrier. To possess.

[0304] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention is such that it is not toxic to the patient. A certain amount of [unclear] effective to achieve the desired therapeutic response in a given patient, composition, and method of administration. It can be modified to obtain the active ingredient. The selected dosage level is based on the referenced text. The specific compositions of the invention, their activity, route of administration, timing of administration, and the elimination of specific compounds used in the invention. Discharge rate, duration of treatment, other drugs used in combination with the specific composition being assisted, Compounds and / or materials, age, sex, weight, condition, and overall health of the patient being treated. It also depends on various pharmacokinetic factors, including factors such as medical history.

[0305] A physician administers the antibody of the present invention, which is incorporated into a pharmaceutical composition, to achieve the desired therapeutic effect. Start at a level below the required level and gradually increase the dosage until the desired effect is achieved. It can be increased. Generally, thrombotic and / or thromboembolic as described herein The dosage of the composition of the present invention, which is effective in treating the disorder, depends on the means of administration, the target site, and the patient's physiological response. This includes the medical condition, other medications being administered, and whether the treatment is preventive or therapeutic. The treatment dosage varies depending on many different factors. The treatment dosage is optimized for safety and efficacy. It is necessary to titrate in this manner. For systemic administration of antibodies, the dose is per kg of host body weight. The dosage ranges from approximately 0.01 to 15 mg. In antibody administration (e.g., subcutaneous administration), the dosage... This can range from 0.1 mg to 5 mg or from 1 mg to 600 mg. For example, in this specification The anti-FXI / FXIa antibodies described are 0.1 mg / kg, 0.2 mg / kg, and 0.3 mg / kg. mg / kg, 0.4mg / kg, 0.5mg / kg, 0.6mg / kg, 0.7mg / k g, 0.8mg / kg, 0.9mg / kg, 1.0mg / kg, 1.1mg / kg, 1. 2mg / kg, 1.3mg / kg, 1.4mg / kg, 1.5mg / kg, 1.6mg / kg, 1.7mg / kg, 1.8mg / kg, 1.9mg / kg, 2.0mg / kg, 2 .1mg / kg, 2.2mg / kg, 2.3mg / kg, 2.4mg / kg, 2.5mg / kg, 2.6mg / kg, 2.7mg / kg, 2.8mg / kg, 2.9mg / kg, 3.0mg / kg, 3.1mg / kg, 3.2mg / kg, 3.3mg / kg, 3.4m g / kg, 3.5mg / kg, 3.6mg / kg, 3.7mg / kg, 3.8mg / kg , 3.9mg / kg, 4.0mg / kg, 4.1mg / kg, 4.2mg / kg, 4.3 mg / kg, 4.4mg / kg, 4.5mg / kg, 4.6mg / kg, 4.7mg / k Administer in doses of g, 4.8 mg / kg, 4.9 mg / kg, or 5.0 mg / kg. This can be done. An example treatment regimen is once every two weeks, or once a month, or 3-6 times. This involves systemic administration once every month. An exemplary treatment regimen is once a week, then once every two weeks. , once every three weeks, once a month, or once every three to six months, or as needed ( This involves systemic administration of PRN.

[0306] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via IV or SC, at a dose of 3 mg / kg. Administer in the specified dose.

[0307] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via IV or SC, at a dose of 10 mg / kJ. Administer in a dose of g.

[0308] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via IV or SC, at a dose of 30 mg / kJ. Administer in a dose of g.

[0309] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via IV or SC, at a dose of 50 mg / kJ. Administer in a dose of g.

[0310] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via IV or SC, at a dose of 100 mg / Administer in a dose of kg.

[0311] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered via, for example, the IV or SC pathway, for 5 minutes. Administer in doses ranging from g to 600 mg.

[0312] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 can be administered, for example, via the IV or SC pathway, for approximately 5 mg, 10mg, 15mg, 20mg, 30mg, 40mg, 50mg, 60mg, 90 mg, 100mg, 120mg, 150mg, 180mg, 200mg, 210mg, 2 40mg, 250mg, 270mg, 300mg, 330mg, 350mg, 360mg , 390mg, 400mg, 420mg, 450mg, 480mg, 500mg, 510 Administer in doses of mg, 540 mg, 550 mg, 570 mg, or 600 mg.

[0313] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via the SC pathway, at a dose of 5 mg. .

[0314] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via the SC pathway, at a dose of 15 mg. ru.

[0315] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via the SC pathway, at a dose of 50 mg. ru.

[0316] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via the SC pathway, at a dose of 150 mg. do.

[0317] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via the SC pathway, at a dose of 300 mg. do.

[0318] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 are administered, for example, via the SC pathway, at a dose of 600 mg. do.

[0319] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 can be administered via, for example, the IV pathway or the SC pathway to aP The average duration of TT was extended by more than double, to 30 days, 35 days, 36 days, and 37 days. 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, Alternatively, administer a dose sufficient to achieve the desired result over a period not exceeding 50 days.

[0320] In certain embodiments, anti-FXI / FXIa antibodies described herein (e.g., NO Antibodies containing VL CDR and VH CDR of V1401, or NOV1401, The antibodies listed in Table 1 can be administered via, for example, the IV pathway or the SC pathway to aP Achieved an extension of more than double the average duration of TT, over a period not exceeding 42 days. Administer a dose sufficient to achieve this.

[0321] Antibodies are typically administered multiple times. The interval between single doses can be weekly, bi-weekly, monthly, or daily. It can be years. The interval also depends on the patient's FXI-binding antibodies and / or FXIa binding. As indicated by measuring the blood levels of compatible antibodies, this can also be irregular. In addition, a doctor may determine an alternative dosing interval, such as monthly or as needed for effective administration. Administer at intervals of 1 to 1. In some methods of systemic administration, the dose is equal to the plasma concentration of the antibody. Adjust to achieve 000 μg / mL or 1-1200 μg / mL, and for several people The method involves adjusting the concentration to achieve 25-500 μg / mL. Alternatively, the antibody is sustained-release. It can also be administered as a compound, in which case low-frequency administration is required. Dosage and The frequency varies depending on the half-life of the antibody and its target in the patient. Generally, in humans Humanized antibodies have a half-life in humans that is longer than that of chimeric antibodies and non-human antibodies. The dosage and frequency of administration vary depending on whether the treatment is preventive or therapeutic. Yes, it is possible. For prophylactic use, relatively low doses are administered over a long period at relatively infrequent intervals. In some cases, the treatment will continue for the rest of the lives of some patients. This means that relatively high doses are required at relatively short intervals until the progression of the disease is reduced or terminated. It is requested, preferably, until the patient shows partial or complete improvement in the symptoms of the disease. Subsequently, the patient can be administered a prophylactic regimen. [Examples]

[0322] The following embodiments are provided to further illustrate the present invention and do not limit its scope. This is not presented for the purpose of determination. Other modifications of the present invention will be readily apparent to those skilled in the art. This would likely be the case, and is also covered by the attached claims.

[0323] Example 1 Panning using a human Fab phage library Multiple panning strategies were employed to select antibodies that recognize human factor XI. Different mutants of the catalytic domain proteins of rabbit factor XI and rabbit factor XIa The therapeutic antibodies used are supplied from commercially available phase display libraries, such as Mo. Using the rphosys HuCAL PLATINUM® library, It was created by selecting clones that bound to factor XI. Lee proposed the HuCAL (registered trademark) concept (Knap) to present Fab on the phage surface. Based on pik et al., 2000, J Mol Biol 296: 57-86), CysDisplay® trademark Using technology (International Publication No. 01 / 05950 pamphlet). Flow of anti-factor XI antibodies. To isolate the body phase, a panning strategy was employed.

[0324] Cross-reactivity analysis Purified Fab is used in ELISA to test for human factor XI (factor XI, factor XIa, etc.). (and the catalytic domain of factor XIa) and the biotiny of the catalytic domain of rabbit factor XIa We investigated the binding of the modified protein to different variants. For this purpose, we used Maxisor p(trademark)(Nunc) 384-well plates are stored at 4°C with 10ug / ml Ne in PBS. The antigen was coated with NeutrAvidin overnight. Above, it was captured via biotin over 30 minutes at room temperature (RT). Different concentrations of Fab were used. The binding is performed using an Attophos fluorescent substrate (Roche: model number 11681982001). It was then conjugated to alkaline phosphatase (diluted to 1:5000). , detected by F(ab)2-specific goat anti-human IgG. Fluorescence emission at 535 nm was The data was recorded by excitation at 430 nm.

[0325] Conversion to IgG and IgG expression To express full-length IgG in CAP-T cells, the variable domain (VH) of the heavy chain is cleaved. The variable domain (VL) fragments of the hilt and light chain were extracted from the Fab expression vector into human IgG1 Subcloning was performed to the appropriate pMorph(registered trademark)_hIg vector at two locations. The amino acid substitutions (D265A and P329A) are introduced into the Fc portion, and any surface relationship Reduced the likelihood of ADCC or CDC caused by FXI. Ranine substitution has been shown to reduce ADCC and CDC (e.g., Idosugie). et al., J. Immunol. 164:4178-4184, 2000; Shields et al., J. Biol. Chem. 276:659 See 1-6604, 2001. Collect the cell culture supernatant 7 days after transfection. After taking the sample and sterilizing and filtering it, the solution was processed using a liquid handling station. The sample was subjected to Tein A affinity chromatography. The sample was eluted in 40 nM NaOH, the pH was neutralized with 1 M Tris buffer, and the sample was filtered sterile. (Pore size of 0.2 μm). Protein concentration was determined by UV spectrophotometry at 280 nm. The purity of IgG was determined and analyzed under denaturation and reduction conditions during SDS-PAGE.

[0326] Example 2 Combined data Surface plasmon resonance (SPR) analysis of FXI catalytic domains SPR measurement is performed using BIACORE, a surface plasmon resonance-based optical biosensor. (Trademark) T200 (BIACORE (Trademark), GE Healthcare, Uppsa The procedure was carried out on la). Series S sensor chip (CM5), immobilization kit, and The regeneration buffer was purchased from GE Healthcare (Uppsala). IgG Alternatively, depending on the ligand format of Fab, two different assay setups can be implemented. The procedure was performed. First, N-hydroxysuccinimide (NHS) and N-(3-dimethylami The surface is activated by (nopropyl)-N-ethylcarbodiimide hydrochloride (EDC). It was converted using the standard amine coupling method (GE Healthcare, Uppsal). a) By using the activated dextran matrix on the CM5 chip, NOV1401-Fab It was covalently attached to the custard. A capture assay was performed on NOV1401-IgG. Then, the goat anti-human IgG-Fc antibody (JIR) was immobilized on the chip at 14,000 RU. The remaining active surface groups were inactivated with ethanolamine (EA). The ligand was immobilized. Prepare reference cells that have not been treated, and set the system in 1x concentration HBS-EP+ buffer (10mM). HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% P2O, pH 7 .4; Equilibrium was achieved using Teknova H8022.

[0327] All binding experiments were performed using HBS-EP+ buffer at 25°C and a flow rate of 50 μL / min. The assay was performed. For the capture assay, NOV1401-IgG was subjected to a RU level of 80. Captured until the end. For reaction rate studies, in the range of 0-200 nM in HBS-EP+ buffer. A dilution series of the FXI catalytic domain was used at the following concentrations. The association time was 120 seconds. The dissociation time was 180 seconds. The surface was treated with a single injection of 10 mM glycine, pH 1.5. The data was regenerated by (a contact time of 60 seconds and a stabilization time of 120 seconds). Processing and k on、 k off , and K D The decision is T200 BiaEvaluat Achieved with ion software version 1.0. Double standard (criteria injection and branding) Apply the deduction for bulk injection to correct for bulk effects and other systematic artifacts. . 1:1 coupling model (R max By applying the settings (which are set globally), I applied the sargram.

[0328] Equilibrium titration (SET) in solution for FXI and FXIa 22 1.6x antigen dilution series (serial 1.6 n dilution) were prepared using sample buffer (0.5% Prepared in PBS (pH 7.4) containing BSA and 0.02% Tween 20. Furthermore, for NOV1401-Fab(huFXI) at a certain concentration, it is set to 200 pM, and hu For FXIa, it was set to 500 pM) or NOV1401 antibody (huFXI and For huFXIa, 10 pM was added to each antigen concentr...

Claims

1. Isolated anti-FXI antibodies and / or antibodies that bind to the catalytic domain of FXI and / or FXIa. Alternatively, isolated anti-FXIa antibodies or fragments thereof.

2. Isolated antibodies that bind to one or more epitopes of anti-FXI and / or FXIa or a fragment thereof, wherein the epitope is Pro410, Arg413, Leu415 , Cys416, His431, Cys432, Tyr434, Gly435, Glu4 37, Tyr472, Lys473, Met474, Ala475, Glu476, Ty r521, Arg522, Lys523, Leu524, Arg525, Asp526, Lys527, Arg548, His552, Ser575, Ser594, Trp59 5. Gly596, Glu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment thereof containing two or more amino acid residues.

3. The aforementioned epitopes are Pro410, Arg413, Leu415, Cys416, Hi s431, Cys432, Tyr434, Gly435, Glu437, Tyr472, Lys473, Met474, Ala475, Glu476, Tyr521, Arg52 2, Lys523, Leu524, Arg525, Asp526, Lys527, Arg 548, His552, Ser575, Ser594, Trp595, Gly596, G four or more of the following meshes: lu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to claim 2, comprising a no acid residue.

4. The aforementioned epitopes are Pro410, Arg413, Leu415, Cys416, Hi s431, Cys432, Tyr434, Gly435, Glu437, Tyr472, Lys473, Met474, Ala475, Glu476, Tyr521, Arg52 2, Lys523, Leu524, Arg525, Asp526, Lys527, Arg 548, His552, Ser575, Ser594, Trp595, Gly596, G Six or more of the following meshes: lu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to claim 2, comprising a no acid residue.

5. The aforementioned epitopes are Pro410, Arg413, Leu415, Cys416, Hi s431, Cys432, Tyr434, Gly435, Glu437, Tyr472, Lys473, Met474, Ala475, Glu476, Tyr521, Arg52 2, Lys523, Leu524, Arg525, Asp526, Lys527, Arg 548, His552, Ser575, Ser594, Trp595, Gly596, G Eight or more of the following meshes: lu597, Arg602, Glu603, and Arg604 An isolated antibody or fragment according to claim 2, comprising a no acid residue.

6. The aforementioned epitopes are Pro410, Arg413, Leu415, Cys416, Hi s431, Cys432, Tyr434, Gly435, Glu437, Tyr472, Lys473, Met474, Ala475, Glu476, Tyr521, Arg52 2, Lys523, Leu524, Arg525, Asp526, Lys527, Arg 548, His552, Ser575, Ser594, Trp595, Gly596, G Claim 2, comprising the residues lu597, Arg602, Glu603, and Arg604. The isolated antibody or fragment described above.

7. The aforementioned epitope consists of the amino acid residues Pro410, Arg413, and Lys527, and L eu415, Cys416, His431, Cys432, Tyr434, Gly435 , Glu437, Tyr472, Lys473, Met474, Ala475, Glu4 76, Tyr521, Arg522, Lys523, Leu524, Arg525, As p526, Arg548, His552, Ser575, Ser594, Trp595, Of the following: Gly596, Glu597, Arg602, Glu603, and Arg604 The isolated antibody or fragment according to claim 2, comprising one or more amino acid residues.

8. The aforementioned epitope consists of the amino acid residues Pro410, Arg413, and Lys527, and L eu415, Cys416, His431, Cys432, Tyr434, Gly435 , Glu437, Tyr472, Lys473, Met474, Ala475, Glu4 76, Tyr521, Arg522, Lys523, Leu524, Arg525, As p526, Arg548, His552, Ser575, Ser594, Trp595, Of the following: Gly596, Glu597, Arg602, Glu603, and Arg604 The isolated antibody or fragment according to claim 2, comprising four or more amino acid residues.

9. The aforementioned epitope consists of the amino acid residues Pro410, Arg413, and Lys527, and L eu415, Cys416, His431, Cys432, Tyr434, Gly435 , Glu437, Tyr472, Lys473, Met474, Ala475, Glu4 76, Tyr521, Arg522, Lys523, Leu524, Arg525, As p526, Arg548, His552, Ser575, Ser594, Trp595, Of the following: Gly596, Glu597, Arg602, Glu603, and Arg604 The isolated antibody or fragment according to claim 2, comprising six or more amino acid residues.

10. Isolated anti-FXI antibodies and / or antibodies that bind to the catalytic domain of FXI and / or FXIa. Alternatively, isolated anti-FXIa antibodies or fragments thereof, and FXI and / or FXI The binding of a to one or more of factor X, factor XIIa, and thrombin An isolated antibody or fragment to block the blockade.

11. Factor IX, Factor XIIa, or thrombin of FXI and / or FXIa, and blocking binding to one or more of the other components of the coagulation pathway, as described in claim 10. The isolated antibody or fragment shown.

12. The binding of one or more of FIX, FXI, and FXIa to platelet receptors An isolated antibody or fragment according to claim 1, which blocks.

13. The isolated antibody according to claim 1 prevents activation of the intrinsic coagulation pathway or the common coagulation pathway. These are fragments.

14. Human FXI protein and / or human FXIa protein, BIACORE (commercial Standard assay (SET) 34 nM or less, measured by an equilibrium titration assay in solution (SET) K below 4 pM as measured by ) Ｄ An isolated antibody or fragment thereof that binds to it.

15. At least one of the CD-Rs listed in Table 1 has at least 90% identity. The isolated antibody or decomposition according to claim 1, comprising at least one complementarity-determining region having Piece.

16. The isolated antibody according to claim 1, comprising CDR1, CDR2, and CDR3 of Table 1 or piece.

17. An isolated variant of the antibody or fragment according to claim 1, wherein the antibody or fragment is as shown in Table 1 The variant includes CDR1, CDR2, and CDR3, and the variant includes CDR1, CDR2, Or, in one of the CDR3 molecules, having at least 1 to 4 amino acid changes, Decentralized mutant.

18. Claim 1, comprising a heavy chain CDR3 selected from the group consisting of Sequence IDs 5 and 25. Isolated antibody or fragment.

20. VH selected from the group consisting of SEQ ID NOs. 9 and 29, or 90% of the same A monogenetic amino acid sequence; selected from the group consisting of SEQ ID NOs: 19 and 39. The claim 1 includes an amino acid sequence having 90% identity with VL, or a sequence having 90% identity thereto. Isolated antibody or fragment.

21. VH selected from the group consisting of SEQ ID NOs. 9 and 29, or 95% of the same A monogenetic amino acid sequence; selected from the group consisting of SEQ ID NOs: 19 and 39. The claim 1 includes an amino acid sequence having 95% identity with VL, or a sequence having 95% identity thereto. Isolated antibody or fragment.

22. VH selected from the group consisting of SEQ ID NOs: 9 and 29, or 97% of the same A monogenetic amino acid sequence; selected from the group consisting of SEQ ID NOs: 19 and 39. The claim 1 includes an amino acid sequence having 97% identity with VL, or the same thereto. Isolated antibody or fragment.

23. The claim 1 includes a variable heavy chain sequence selected from the group consisting of SEQ ID NOs: 9 and 29. Isolated antibody or fragment.

24. The variable light chain sequence selected from the group consisting of sequence numbers 19 and 39, as described in claim 1. The isolated antibody or fragment shown.

25. Variable heavy chains selected from the group consisting of SEQ ID NOs: 9 and 29; and SEQ ID NOs: 19 and The isolated antibody according to claim 1, comprising a variable light chain sequence selected from the group consisting of 39 or piece.

26. If an antibody or fragment containing the variable heavy chain sequence of SEQ ID NO: 9 and the variable light chain sequence of SEQ ID NO: 19 is used, Furthermore, an antibody or fragment containing the variable heavy chain sequence of SEQ ID NO: 29 and the variable light chain sequence of SEQ ID NO: 39 An isolated antibody or fragment according to claim 1, selected from the group.

27. CDR1 of the heavy chain variable region selected from the group consisting of SEQ ID NO: 46; consisting of SEQ ID NO: 4 CDR2 selected from the group; CDR3 selected from the group consisting of 5; sequence number 33 CDR1 of the light chain variable region selected from the group; C selected from the group consisting of sequence number 14. Claim 1 includes CDR3 selected from the group consisting of DR2 and Sequence ID No.

15. Isolated antibody or fragment.

28. CDR1 of the heavy chain variable region selected from the group consisting of SEQ ID NOs: 3 and 23; SEQ ID NO: 4 and selected from the group consisting of 24; selected from the group consisting of 5 and 25 CDR3; CDR1 of the light chain variable region selected from the group consisting of sequence numbers 13 and 33; CDR2 selected from the group consisting of SEQ ID NOs: 14 and 34, and SEQ ID NOs: 15 The isolated antibody or fragment according to claim 1, comprising CDR3 selected from the group consisting of 35. 。

29. CDR1 of the heavy chain variable region selected from the group consisting of SEQ ID NOs: 6 and 26; SEQ ID NO: 7 and selected from the group consisting of 27; CDR2 selected from the group consisting of 8 and 28 CDR3; CDR1 of the light chain variable region selected from the group consisting of sequence numbers 16 and 36; CDR2 selected from the group consisting of SEQ ID NOs: 17 and 37, and SEQ ID NOs: 18 and The isolated antibody or fragment according to claim 1, comprising CDR3 selected from the group consisting of 38. 。

30. CDR1 of the heavy chain variable region of sequence number 3; CDR2 of the heavy chain variable region of sequence number 4; sequence number CDR3 of the heavy chain variable region of sequence 5; CDR1 of the light chain variable region of sequence number 13; sequence number 14 The claim includes CDR2 of the light chain variable region of sequence number 15, and CDR3 of the light chain variable region of sequence number 15. The isolated antibody or fragment described in item 1.

31. CDR1 of the heavy chain variable region of SEQ ID NO: 23; CDR2 of the heavy chain variable region of SEQ ID NO: 24; CDR3 of the heavy chain variable region of sequence number 25; CDR1 of the light chain variable region of sequence number 33; sequence number Includes CDR2 of the light chain variable region of sequence 34; and CDR3 of the light chain variable region of sequence number 35. The isolated antibody or fragment according to claim 1.

32. CDR1 of the heavy chain variable region of sequence number 6; CDR2 of the heavy chain variable region of sequence number 7; sequence number CDR3 of the heavy chain variable region of sequence number 8; CDR1 of the light chain variable region of sequence number 16; sequence number 17 The claim includes CDR2 of the light chain variable region of sequence number 18, and CDR3 of the light chain variable region of sequence number 18. The isolated antibody or fragment described in item 1.

33. CDR1 of the heavy chain variable region of SEQ ID NO: 26; CDR2 of the heavy chain variable region of SEQ ID NO: 27; CDR3 of the heavy chain variable region of sequence number 28; CDR1 of the light chain variable region of sequence number 36; sequence number Includes CDR2 of the light chain variable region of sequence 37; and CDR3 of the light chain variable region of sequence number 38. The isolated antibody or fragment according to claim 1.

34. A medical device comprising an antibody or fragment thereof as described in one of the above claims and a pharmaceutically acceptable carrier. A pharmaceutical composition.

35. A claim that binds to the same epitope as the isolated antibody or fragment described in any of the above claims. The isolated antibody or fragment described in item 1.

36. Regarding binding to human FXI protein and / or human FXIa protein, The isolated antibody or fragment described in any of the claims, which competes with the isolated antibody or fragment described in claim 1. These are fragments.

37. A single unit according to claim 1, selected from the group consisting of NOV1090 and NOV1401. Antibody release or fragment.

38. A method for treating thromboembolic disorders, comprising an effective amount of the antimicrobial agent according to any of the claims. A pharmaceutical composition containing a body or fragment is administered to a subject suffering from a thromboembolic disorder. A method that includes a top.

39. The aforementioned subject has one of the following conditions associated with atrial fibrillation: ischemic stroke or deep vein thrombosis. The method according to claim 38, wherein the person is suffering from multiple conditions.

40. The person according to claim 38, wherein the subject is suffering from an ischemic stroke associated with atrial fibrillation. Law.

41. A method for treating thromboembolic disorders, comprising an effective amount of the antimicrobial agent according to any of the claims. A pharmaceutical composition containing a body or fragment, in combination with statin therapy, is used to treat thromboembolic disorders. A method comprising the step of administering to a target.

42. A pharmaceutical product comprising the antibody described in any of the above claims.

43. A nucleic acid encoding one or more of the antibodies described in any of the above claims.

44. A vector comprising the nucleic acid described in claim 43.

45. A host cell comprising the vector according to claim 44.

46. When it binds to the active FXI (FXIa) catalytic domain, it conforms to FXIa. The four N-terminal residues, loops 145, 188, and 220, are activated comfort In contrast to conformations, inert conformations are shifted and / or irregular. The isolated antibody or fragment according to claim 1, which is transformed into n.

47. When bound to FXI, the FXI catalytic domains, loops 145, 188, and 220 Adopting an active conformation that is as regular as the structure within the FXIa catalytic domain. An isolated antibody or fragment according to claim 1, which prevents the following.

48. When bound to FXI, the FXI catalytic domain consists of four residues at the N-terminus, loop 145, and 1. 88 and 220 are as regular as within the structure of the FXIa catalytic domain, active conformation The isolated antibody or fragment according to claim 1, which prevents the formation of a malformation.

49. When it binds to FXI, it induces conformational changes within the zymogen structure, and FXIa Inhibitory FXI conformation observed when binding to is closely related to inhibitory F By further introducing the XI conformation, the FXI catalytic domain becomes active. The isolated antibody or fragment according to claim 1, which prevents formation.

50. The catalyst domaine is bound to FXI and / or FXIa. When the antibody forms an antigen complex, the activity of loops 145, 188, and 220 The catalytic domain of factor XI (FXIa) compared to the uncomplexed structure An isolated antibody or fragment according to claim 1, which causes loss of pharmacokinetics and / or orientation.

51. The catalyst domaine is bound to FXI and / or FXIa. When an antibody forms an antigen complex, the four residues at the N-terminus, loop 145, 188, The uncomplexed structure of the catalytic domain of the active factor XI (FXIa) of 220 The method described in claim 1 causes a shift and / or loss of orientation compared to the construction. Isolated antibody or fragment.

52. It binds to the active FXI(FXIa) and to the FXI(FXIa) catalytic domain, and its conjugate Loops 145, 188, and 220 compare the conformation to the active conformation. It shifts and / or changes into an inert conformation in which orientation is lost. The isolated antibody or fragment according to claim 1.

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