Administration of polyomavirus neutralizing antibodies

Abstract

This invention provides a dosage regimen for the treatment of BK or JC polyomavirus infection in human subjects. [Solution] The administration regimen includes the steps of (a) parenterally administering a dose of an antibody or antigen-binding fragment that specifically binds to the VP1 protein of the polyomavirus to the subject; (b) measuring the serum or tissue concentration of the antibody or antigen-binding fragment in the subject; and (c) measuring the serum or tissue trough concentration (C trough The administration regimen includes the step of administering a further dose of the antibody or its antigen-binding fragment before the concentration drops to less than approximately 3-860 μg / mL, wherein the concentration of the antibody or its antigen-binding fragment in the serum or tissue during the treatment is reduced to the C trough Maintain a higher position.

Description

[Technical Field]

[0001] Cross-reference of related applications This application, dated June 12, 2020, is incorporated herein by reference in its entirety. We claim priority from U.S. Provisional Patent Application No. 63 / 038,433, filed in [location].

[0002] Statements regarding sequence listings The sequence listing associated with this application is provided in text format instead of paper copy, and is available for reference. This specification incorporates the following. The name of the text file containing the sequence listing is AMP The file is X_003_01WO_ST25.txt. The text file is approximately 24KB. It was created on June 4, 2021, and is scheduled to be submitted electronically via EFS-Web. be.

[0003] background Technical field This disclosure relates to polyomavirus neutralizing antibodies for treating polyomavirus infection. This relates to dosing regimens and related methods and pharmaceutical compositions.

[0004] Description of prior art Polyomaviruses such as BK virus and JC virus are widely distributed throughout the world. The prevalence is approximately 75-90%. After the initial infection, the virus is usually expelled from most people. Although it remains dormant, immunosuppressive therapy to prevent graft rejection leads to immunosuppression. Reactivation is observed in immunocompromised patients, including transplant patients. It's possible.

[0005] For example, in kidney transplant recipients, BK virus reactivation occurs in BK virus blood. BK can lead to this condition, and it is the main cause of irreversible kidney injury and graft failure. V-associated nephropathy may occur. The current standard treatment is to reduce immunosuppression, which is metastatic. There is a considerable risk of immune system rejection of the grafted organ. Also, hematopoietic stem cell transplantation (HCT) recipe BK virus reactivation in entrenches can lead to BKV-associated hemorrhagic cystitis (lasting several weeks to several months). Hemorrhagic cystitis (a condition accompanied by severe pain that can persist and lead to prolonged hospitalization). Current treatments for this condition are generally symptomatic, and include narcotic analgesics, hydration, Other treatments include diuresis. Many patients also require bladder irrigation, clot removal, blood transfusions, and stent placement. , and nephrostomy is required. Currently, there are no approved treatments for BK virus infection. stomach.

[0006] MAU868 is a human monoclonal antibody that potently neutralizes all four BKV serotypes. (Immunoglobulin G, IgG1 / λ isotype subclass) (e.g., P8D) (See International Publication No. 2017 / 046676, as item 11). The viral capsid protein (VP) is involved in binding to target cells and promoting infection. 1) Recognizes the three-dimensional structure epitope above. MAU868 also recognizes the closely related JC virus (advance It has neutralizing activity against the cause of multifocal leukoencephalopathy. This neutralizing antibody is used in kidney transplant patients. In this context, by avoiding the most frequent cause of viral allograft loss, complications To minimize the burden and to significantly simplify the immunosuppressive therapy regimens that clinicians currently face. This was possible. Similarly, BK virus-associated hemorrhagic cystitis in HCT recipients Treatment or prevention of complications with significant morbidity and mortality rates, as well as acute graft-versus-host complications, are important. The disease could be eliminated or reduced.

[0007] However, the need to optimize the dosing of polyomavirus neutralizing antibodies is It exists in the field of technology. [Overview of the project]

[0008] Brief summary Embodiments of this disclosure relate to BK or JC polyoma in human subjects requiring it. A regimen for the treatment of viral infections, (a) an antibody or antigen that specifically binds to the VP1 protein of the polyomavirus A step of administering the combined fragment dose to the target patient parenterally; (b) Measuring the serum or tissue concentration of the antibody or its antigen-binding fragment in the subject. Step; and (c) trough concentration in serum or tissue (C trough ) is approximately 3-860 μg Administer additional doses of the antibody or its antigen-binding fragment before the dosage drops below less than / mL. Step The administration regimen includes, and the concentration of the antibody or its antigen-binding fragment in serum or tissue, Throughout the procedure, C trough Includes administration regimens to maintain a higher level.

[0009] In some embodiments: (i)(c) C trough This is about plasma, and the administration regimen is blood Serum C trough Before the dose drops to less than approximately 150-860 μg / mL, further doses may be required. The step includes administering; (ii)(c) C trough This is about kidney tissue, and the administration regimen is C in renal tissue troughBefore it drops to less than about 23.5 to 120 μg / mL, including the step of administering a further dosage; or (c) in (iii) trough is for bladder tissue, and the dosing regimen includes the step of administering a further dosage before C in the bladder tissue trough drops to less than about 3 to 10 μg / mL.

[0010] In some embodiments, the antibody or antigen-binding fragment thereof comprises the complementarity-determining regions (CDRs) V H CDR1, V H CDR2, and V H CDR3 sequences of the heavy-chain variable (V H ) region; and the V L CDR1, V L CDR2, and V L CDR3 sequences of the light-chain variable (V ) region, L as well as variants thereof that specifically bind to the VP1 protein including.

[0011] In some embodiments: The V H region comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, 98 %, 99%, or 100% identical to SEQ ID NO: 12, and optionally, the V H sequence optionally has one or more of the alterations selected from one, two, three, four, five, or six of V 5Q, G9P, T10G, N30S, N30K, and N30Q in the framework region; and The V region comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, 98 L % identical to SEQ ID NO: 13, and Contains %, 99%, or 100% identical sequences, and optionally, V L The array consists of 1, 2, The framework has three, four, five, or six variations.

[0012] In some embodiments, V H The region contains or consists of sequence number 12. Or essentially become V L The region includes or consists of sequence number 13, Then it becomes essential.

[0013] In some embodiments, the administration regimen is determined by the BK virus genotype in the subject. The procedure includes the step of identifying the JC virus. In some embodiments, polyoma Viral infections are selected from one or more genotypes I, II, III, and IV. This includes BK virus, or polyomavirus infections include JC virus.

[0014] In some embodiments, the subjects are immunocompromised. In some embodiments, the subject is a transplantation procedure, optionally organ transplantation or cell-based I am about to undergo, and am currently undergoing, a (cell-based) transplant procedure, and I have received it. In some embodiments, the transplant procedure involves kidney transplantation and hematopoietic cell transplantation. Selected from cyst transplantation (HCT). In some embodiments, the target is BK virus Associated nephropathy, BK virus-associated hemorrhagic cystitis, and JC virus-associated progressive multifocal leukoencephalopathy You have, or are at risk of having, a condition selected from the list of symptoms.

[0015] In some embodiments, the dosage in (a) is approximately 1 to approximately 100 mg / kg. Alternatively, approximately 10-30 mg / kg, or approximately 1 mg / kg, 2 mg / kg, 3 mg / kg, 4mg / kg, 5mg / kg, 6mg / kg, 7mg / kg, 8mg / kg, 9mg / k g, 10mg / kg, 15mg / kg, 20mg / kg, 25mg / kg, 30mg / k g, 35mg / kg, 40mg / kg, 45mg / kg, 50mg / kg, 60mg / k g, 70 mg / kg, 80 mg / kg, 90 mg / kg, or 100 mg / kg The dosage is optionally administered intravenously or subcutaneously. In some embodiments, The additional dosage in (c) may be the same as or different from the dosage in (a), and is optional. The options are approximately 1 to 100 mg / kg or approximately 10 mg / kg to 30 mg / kg, or Approximately 1mg / kg, 2mg / kg, 3mg / kg, 4mg / kg, 5mg / kg, 6mg / kg, 7mg / kg, 8mg / kg, 9mg / kg, 10mg / kg, 15mg / kg, 20mg / kg, 25mg / kg, 30mg / kg, 35mg / kg, 40mg / kg, 45mg / kg, 50mg / kg, 60mg / kg, 70mg / kg, 80mg / kg, The dosage is 90 mg / kg or 100 mg / kg, and is optional. The dosage is administered intravenously or It is administered subcutaneously.

[0016] In some embodiments, polyomavirus infection is caused by BK virus genotype I This includes, and here, C trough Tissue concentrations in the region, and optionally, renal tissue concentrations, of the antibody or its antigen. EC of synthetic fragments 50 From approximately 2618 times to approximately 3775 times, up to approximately 13,061 times (fr The range is approximately 2618 to approximately 3775 to approximately 13,061 (fold); or C trough Tissue concentrations in the region, and optionally, bladder tissue concentrations, of the antibody or its antigen. EC of binding fragments 50 The range is approximately 500 times to 692 times, and up to 1000 times. , EC 50 The concentration is approximately 0.009 ± 0.010 μg / mL.

[0017] In some embodiments, polyomavirus infection is BK virus genotype II This includes, and here, C trough Tissue concentrations in the region, and optionally, renal tissue concentrations, of the antibody or its antigen. EC of synthetic fragments 50 From approximately 589 times to approximately 849 times, and up to approximately 2942 times (for example) , approximately 600x, 700x, 800x, 900x, 1000x, 1500x, 2000x, (2500 times, 3000 times) and; or C trough Tissue concentrations in the region, and optionally, bladder tissue concentrations, of the antibody or its antigen. EC of binding fragments 50 The range is approximately 100 times to 156 times, and up to 200 times. EC 50 The concentration is approximately 0.040 ± 0.025 μg / mL.

[0018] In some embodiments, polyomavirus infection is BK virus genotype II I include, and here, C trough Tissue concentrations in the region, and optionally, renal tissue concentrations, of the antibody or its antigen. EC of synthetic fragments 50 The range is approximately 253 times to approximately 365 times, and up to approximately 1265 times; or C troughTissue concentrations in the region, and optionally, bladder tissue concentrations, of the antibody or its antigen. EC of binding fragments 50 From approximately 50 times to approximately 67 times, up to approximately 100 times (for example, approximately 50x, 55x, 60x, 65x, 70x, 75x, 80x, 85x, 90x, 95x, (or 100 times), EC 50 The concentration is approximately 0.093 ± 0.057 μg / mL.

[0019] In some embodiments, polyomavirus infection is BK virus genotype IV This includes, and here, C trough Tissue concentrations in the region, and optionally, renal tissue concentrations, of the antibody or its antigen. EC of synthetic fragments 50 The range is approximately 1122 times to approximately 1618 times, and up to approximately 5604 times. ri; or C trough Tissue concentrations in the region, and optionally, bladder tissue concentrations, of the antibody or its antigen. EC of binding fragments 50 The range is approximately 100 times to 297 times, and up to 500 times. EC 50 The concentration is approximately 0.021 ± 0.020 μg / mL.

[0020] In some embodiments, polyomavirus infection includes the JC virus, C t rough The tissue concentration in the antibody or its antigen-binding fragment is the EC 50 at least From approximately 29 times to approximately 110 times, up to approximately 158 times, up to approximately 547 times (at least about 29 to about 110 to about 158 ​​to about 547 (fold), EC 50 It is approximately 0.215±0 The concentration is 0.130 μg / mL.

[0021] In some embodiments, the time between (a) and (b) is approximately or at least approximately 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 1 3rd, 14th, 14th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 2 3rd, 24th, 25th, 26th, 27th, 28th, 29th, or 30th, or 1 Weeks, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, or 1 month, 2 months, Or 3 months, or approximately 1 day or less, 2 days or less, 3 days or less, 4 days or less, 5 days or less, 6 days or less , 7 days or less, 8 days or less, 9 days or less, 10 days or less, 11 days or less, 12 days or less, 13 days or less, 14 days or less, 14 days or less, 16 days or less, 17 days or less, 18 days or less, 19 days or less, 20 days or less Below, 21 days or less, 22 days or less, 23 days or less, 24 days or less, 25 days or less, 26 days or less, 27 Less than a day, 28 days or less, 29 days or less, or 30 days or less, or 1 week or less, 2 weeks or less, 3 weeks or less, 4 weeks or less, 5 weeks or less, 6 weeks or less, 7 weeks or less, or 8 weeks or less, or 1 month It is less than one month, less than two months, or less than three months. A particular dosing regimen is (b) In elephants, serum or tissue levels were observed for approximately 3, 4, 5, 6, 7, 8, and 9 days. 10th, 11th, 12th, 13th, 14th, 14th, 16th, 17th, 18th, 19th , 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th , or once every 30 days, or approximately every 1, 2, 3, 4, 5, 6, 7 weeks, or The test is performed 1-3 times every 8 weeks, or 1-6 times every 1, 2, or 3 months. Includes a stop. In some embodiments, the time between (a) and (c) is about or slightly longer. At least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12th, 13th, 14th, 14th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, or 30th, Or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, or 1 month, Two months, or three months, or approximately one day or less, two days or less, three days or less, four days or less, five days or less , 6 days or less, 7 days or less, 8 days or less, 9 days or less, 10 days or less, 11 days or less, 12 days or less, 1 3 days or less, 14 days or less, 14 days or less, 16 days or less, 17 days or less, 18 days or less, 19 days or less , 20 days or less, 21 days or less, 22 days or less, 23 days or less, 24 days or less, 25 days or less, 26 days Below are examples of periods of 27 days or less, 28 days or less, 29 days or less, or 30 days or less, or 1 week or less. Less than 2 weeks, less than 3 weeks, less than 4 weeks, less than 5 weeks, less than 6 weeks, less than 7 weeks, or less than 8 weeks. Or it may be less than one month, less than two months, or less than three months. In some embodiments The circulating concentration of the antibody or its antigen-binding fragment in the serum is measured. at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% of (tion), 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 2 0% or more penetrate the infected organ. In some embodiments... The infected organs are the bladder, kidneys, or central nervous system (CNS).

[0022] In some embodiments, the mean clearance (me) of the antibody or its antigen-binding fragment The clearance is approximately 0.0760-0.0996 mL / day / kg. In the application method, the mean volume of distribution of the antibody or its antigen-binding fragment is called the mean volume of distribution. The saturation level is approximately 49.8–81.9 mL / kg.

[0023] To treat BK or JC polyomavirus infections in human subjects who require it. A method for which an antibody specifically binds to the VP1 protein of polyomavirus The procedure includes a step of administering the antigen-binding fragment parenterally, and the dosage is Approximately 10-30 mg / kg, or approximately 10 mg / kg, 11 mg / kg, 12 mg / kg , 13mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17mg / kg , 18mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22mg / kg , 23mg / kg, 24mg / kg, 25mg / kg, 26mg / kg, 27mg / kg Methods that involve 28 mg / kg, 29 mg / kg, or 30 mg / kg are also included. ru.

[0024] In some embodiments, the antibody or its antigen-binding fragment is Complementarity Determination Region (CDR) V corresponding to sequence numbers 6-8. H CDR1, V H CDR2, oh bi V H Heavy chain variable (V) containing the CDR3 sequence H ); and V corresponding to sequence numbers 9-11 L CDR1, V L CDR2, and V L Includes CDR3 sequence Variable light chain (V L )region, as well as those variants that specifically bind to the VP1 protein Includes.

[0025] In some embodiments: V H The region is at least 80%, 85%, 90%, 95%, 97%, and 98% of sequence number 12. Contains %, 99%, or 100% identical sequences, and optionally, V H The array can be selected as V. Choose one or more from 5Q, G9P, T10G, N30S, N30K, and N30Q. The framework has one, two, three, four, five, or six possible changes to choose from within its domain. shi; and V L The region is sequence number 13 and at least 80%, 85%, 90%, 95%, 97%, 98%. Contains %, 99%, or 100% identical sequences, and optionally, V L The array consists of 1, 2, The framework has three, four, five, or six variations.

[0026] In some embodiments, V H The region contains or consists of sequence number 12. Or essentially become V L The region includes or consists of sequence number 13, Then it becomes essential.

[0027] In some embodiments, polyomavirus infection is classified as genotype I, II, III , and BK viruses selected from one or more of IV. Several embodiments In this context, polyomavirus infections include JC virus.

[0028] In some embodiments, the subjects are immunocompromised. The target group is those who undergo transplant procedures, either organ transplantation or cell-based transplantation procedures of their choice. They are about to receive, are currently receiving, or have received. Several implementation forms In this context, the transplant procedure is selected from kidney transplantation and hematopoietic stem cell transplantation (HCT). In several embodiments, the target is BK virus-associated nephropathy, BK virus-associated hemorrhagic bladder Having or having a condition selected from inflammation and JC virus-associated progressive multifocal leukoencephalopathy. There is a risk of that happening.

[0029] In some embodiments, the circulating concentration of the antibody or its antigen-binding fragment in the serum is small. At least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 1 2%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%, also The virus penetrates further into the infected organ. In some embodiments, the infected organ is the bladder. or is the kidney. In some embodiments, the average of the antibody or its antigen-binding fragment The clearance is approximately 0.0760–0.0996 mL / day / kg. Several implementations In this state, the mean distribution volume of an antibody or its antigen-binding fragment is approximately 49.8–81.9 m³. The dosage is L / kg. In some embodiments, the dosage is 10-30 mg / kg. When measured in an in vitro or ex vivo viral assay, BK or JC positivity Optimal neutralizing activity (IC) of target serum against riomavirus 50 ) gives.

[0030] In some embodiments, the administration regimen or method involves BK or J in the subject. C viremia and / or viruria, optionally, with control or antibody or Compared to the pre-treatment state with antigen-binding fragments, approximately or at least approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 4 00%, 500%, 1000%, 2000%, 3000%, 4000%, or 5000 Reduce by %, or more.

[0031] In some embodiments, the administration regimen or method may be one or more in the subject. A number of BK or JC virus-related symptoms, optional, blurred vision or other Visual changes, brown or red urine, painful urination, decreased kidney function, difficulty urinating, cough, common cold, difficulty breathing. One of the following symptoms may be selected from difficulty breathing, fever, muscle pain, muscle weakness, and / or seizures. It reduces kidney inflammation such as ureteral stricture or interstitial nephritis, and is used selectively for multiple symptoms. This improves. In some embodiments, the administration regimen or method is histidine, sugar Saccharides (saccharides), optional sucrose, and polyols, optional In a pharmaceutical composition containing polysorbate, an antibody or its antigen-binding fragment is administered. Includes steps.

[0032] Formulated for parenteral administration at a dosage of approximately 10-30 mg / kg, and each is distributed Complementarity Determination Region (CDR) V in columns 6-8 H CDR1, V H CDR2, and V H CD Heavy chain variable (V) containing the R3 sequence H ), and V of sequence numbers 9-11, respectively. L CDR1, V L CDR2, and V L Light chain variable (V) containing CDR3 sequence L ) an antibody or including the region Its antigen-binding fragment; and Histidine, sugars, optionally sucrose, and polyols, optionally polysorbates. pharmaceutically acceptable carriers including Pharmaceutical compositions containing the same are also included.

[0033] In some embodiments, V H The region is at least 80% to 85% of sequence number 12. , containing 90%, 95%, 97%, 98%, 99%, or 100% identical sequences, and any selection In the selection, V H The sequences are V5Q, G9P, T10G, N30S, N30K, and One, two, three, four, five, or six N30Q models can be selected from one or more options. The framework has undergone changes; and V L The region is sequence number 13 and at least 80%, 85%, 90%, 95%, 97%, 98%. Contains %, 99%, or 100% identical sequences, and optionally, V L The array consists of 1, 2, The framework has three, four, five, or six variations.

[0034] In some embodiments, the antibody or its antigen-binding fragment is approximately 10 mg / kg. 11mg / kg, 12mg / kg, 13mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17mg / kg, 18mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22mg / kg, 23mg / kg, 24mg / kg, 25mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, or 30 mg / It is formulated for parenteral administration in doses of kg. Certain compositions are for intravenous administration. It is formulated for administration. Some compositions are formulated for subcutaneous administration. .

[0035] Some embodiments describe using BK or JC polyoma in human subjects where it is needed. The pharmaceutical compositions described herein for use in treating viral infections Includes. In some embodiments, polyomavirus infection is genotype I, II, I Includes BK viruses selected from one or more of II and IV. Several implementations Morphologically, polyomavirus infections include the JC virus. In some embodiments... The subjects have a weakened immune system. In some embodiments, the subjects undergo a transplant procedure, optional They are about to undergo an organ transplant or a cell-based transplant procedure of their choice. They are in the process of doing or have done. In some embodiments, the transplant procedure involves the kidney Selected from organ transplantation and hematopoietic stem cell transplantation (HCT). In some embodiments, Elephants are associated with BK virus-associated nephropathy, BK virus-associated hemorrhagic cystitis, and JC virus-associated diseases. You have or are at risk of having a condition selected from progressive multifocal leukoencephalopathy. [Brief explanation of the drawing]

[0036] [Figure 1] Figure 1 shows the time course of exposure to MAU868 after intravenous or subcutaneous administration. [Figure 2] Figure 2 shows the ex vivo serum neutralizing activity of MAU868 from patient serum relative to its serum concentration. BKVEC50 = the volume of serum required to neutralize 50% of an in vitro infection. [Modes for carrying out the invention]

[0037] Detailed explanation Unless otherwise specified, all technical and scientific terms used herein are the same as those used in this disclosure. This has the same meaning as is generally understood by those skilled in the art to which it belongs. Any method, material, etc. that is similar to, or equivalently similar to, or equivalent to, what is described. Compositions, reagents, and cells may be used in the conduct or testing of the subject matter of this disclosure, but preferably Methods and materials are described. All publications and references cited in this specification. The contribution (for example, patents and patent applications, but not limited to them) is as if Each individual publication or reference is specifically and individually cited, as if fully indicated. The whole is incorporated herein by reference as if it were incorporated herein by reference It is included. Any patent application claiming priority from this application is also included in the publications and references. The references are incorporated into this specification by reference in the format described above. .

[0038] Recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., Standard techniques can be used for electroporation (lipofection). Application and refining techniques are carried out in accordance with the manufacturer's specifications or are generally practiced in the art. These may be carried out as done or as described herein. The related techniques and procedures described above are generally well-known and common in the art. In accordance with the law, as well as various general and It may be implemented as described in more specific references. Specific definitions are provided. Unless otherwise specified, the molecular biology, analytical chemistry, synthetic organic chemistry, and Nomenclature used in relation to medicine and medicinal chemistry, as well as related experimental procedures and techniques, This is well known and commonly used in the field of recombinant technology. Medicine, molecular biology, microbiology, chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, and delivery. Furthermore, standard techniques may be used in the treatment of patients.

[0039] For the purposes of this disclosure, the following terms are defined below.

[0040] The articles "a" and "an" refer to one of the grammatical objects of the article or In this specification, it is used to refer to more than one (i.e., at least one). For example, And, "an element" is "one element", "one or Multiple elements (one or more elements), and / or "at least one element (at l Includes "east one element".

[0041] "Approximately" refers to the quantity, level, value, number, frequency, percentage, or dimension of a reference. 10%, 9%, 8%, 7%, 6%, 5% of size, amount, weight, or length A quantity, level, value, number, or frequency that changes by approximately %, 4%, 3%, 2%, or 1%. It can mean percentage, dimension, size, amount, weight, or length.

[0042] The term "antigen" refers to a substance that can be bound by selectively binding substances such as antibodies, and furthermore... In addition, animals are used to produce antibodies that have the ability to bind to the epitope of that antigen. An antigen refers to a molecule or part of a molecule that can be identified. It may have a peculiarity. As used herein, the term "antigen" means that, under appropriate conditions, it is the substance of the object. It can induce an immune response to quality, and can react with the products of that immune response. It contains substances that can trigger an antigen, such as an antibody (humoral immune response) or a sensitized T lymphocyte. The antigen can be recognized by (helper T cells or cell-mediated immune response) or both. Soluble substances, such as toxins and foreign proteins, or particulate matter, such as bacteria and tissue particles. It may be a cell; however, the protein known as the antigenic determinant (epitope) In this case, only a portion of the polysaccharide molecule binds to a specific receptor on the antibody or lymphocyte. In other words, the term "antigen" refers to a substance to which an antibody binds, regardless of whether the substance is immunogenic or not. or any substance for which antibodies are desired. For such antigens, antibodies are any It can be identified by recombinant methods, regardless of the immune response.

[0043] An "antagonist" is a substance or molecule that interferes with or alters the physiological effects of another active substance or molecule. It refers to a biological structure or chemical substance that reduces something. In some cases, the antagonist is It specifically binds to the other active substance or molecule. Complete and partial antagonists It is included.

[0044] An "agonist" is a substance that increases or enhances the physiological effects of another active substance or molecule. It refers to a physical structure or chemical substance. In some cases, an agonist is the other active substance or It specifically binds to molecules. This includes complete and partial agonists.

[0045] As used herein, the term "amino acid" refers to naturally occurring amino acids and naturally occurring amino acids This is intended to mean both amino acids that are not present, as well as amino acid analogs and mimetic compounds. Naturally occurring amino acids include 20 (L)- amino acids that are used in protein biosynthesis. Amino acids, plus others, for example, 4-hydroxyproline, hydroxylysine, desmo Examples include syn, isodesmosine, homocysteine, citrulline, and ornithine. Amino acids that do not exist in nature include, for example, (D)-amino acids, which are known to those skilled in the art. norleucine, norvaline, p-fluorophenylalanine, ethionine, and the Other similar substances can be mentioned. Amino acid analogs include naturally occurring amino acids and natural Examples of such modifications include altered amino acids that do not exist in the original amino acid. This can be achieved by substitution or replacement of chemical groups and parts, or by derivatization of amino acids. It is possible. Amino acid mimetic compounds may have, for example, the charge and charge space specific to the reference amino acid. Examples of organic structures that exhibit functionally similar properties include syng, for instance, arginine ( Organic structures that mimic Arg (or R) are located in a similar molecular space and are naturally occurring. It has a positively charged portion that has the same degree of mobility as the e-amino group of the side chain of the Arg amino acid. The mimetic also features optimal spacing and charge interaction of amino acids or amino acid functional groups. It includes a structure that is constrained in order to maintain its function. Those skilled in the art will know what structure is functionally equivalent. It is known or possible to determine whether they constitute an acid analog and an amino acid mimetic. ru.

[0046] As used herein, the term “antibody” means intact polyclonal or monoclonal Not only the whole antibody, but also its fragments (dAb, Fab, Fab', F(ab')2, Fv), Single-stranded (ScFv), its synthetic variant, naturally occurring variant, required features Fusion proteins containing antibody moieties with heterozygous antigen-binding fragments, humanized antibodies, and chimeric antibodies. an immunoglobulin containing an antigen-binding site or fragment (epitope recognition site) of the required specificity. This also includes any other modified stereochemistry of the robulin molecule. Antibodies (and their antigen-binding properties) Certain features and properties of the fragments are described in more detail herein.

[0047] Antibodies or antigen-binding fragments can be of essentially any type. As is well known, antibodies are immunoglobulins that target immune checkpoint molecules and other targets. Specific binding occurs through at least one epitope recognition site located in the variable region of the molecule. It is an immunoglobulin molecule with the ability to combine.

[0048] As used herein, the term “antigen-binding fragment” means an immunoglobulin that binds to the antigen of interest. Polypeptide fragment containing at least one CDR of robulin heavy chain and / or light chain This refers to the antigen-binding fragment of the antibody described herein, which binds to the target molecule. V derived from the binding antibody H and V L One, two, three, four, five, or all six elements in the array This may include a CD-R.

[0049] The binding properties of antibodies and their antigen-binding fragments are determined by methods well known in the art ( Quantification should be performed using the method described in Davies et al., Annual Rev. Biochem. 59:439-473, 1990. This is possible. In some embodiments, the antibody or its antigen-binding fragment can target a molecule. For example, the VP1 protein or its epitope or complex, and approximately ≤10 -7 M or about 10 -7 From M to about 10 -8 The equilibrium dissociation constant (an equilibri) is in the range up to M. um dissociation constant that is about or ranges from about < 10 -7 M to about 10 - 8 M) specifically binds. In some embodiments, the equilibrium dissociation constant is approximately ≤ 10 -9 It is M, or about 10 -9 From M to approximately ≤10 -10 The range is up to M (is abo ut or ranges from about < 10 -9 M to about < 10 -10 M). In a particular exemplary embodiment In this case, the antibody or its antigen-binding fragment is approximately or at least approximately 0.01 nM, 0.0 5nM, 0.1nM, 0.2nM, 0.3nM, 0.4nM, 0.5nM, 0.6nM, 0.7nM, 0.8nM, 0.9nM, 1nM, 2nM, 3nM, 4nM, 5nM, 6n M, 7nM, 8nM, 9nM, 10nM, 11nM, 12nM, 13nM, 14nM, 1 5nM, 16nM, 17nM, 18nM, 19nM, 20nM, 21nM, 22nM, 2 3nM, 24nM, 25nM, 26nM, 27nM, 28nM, 29nM, 30nM, 4 0 nM, or 50 nM, or less than approximately 0.01 nM, less than 0.05 nM, 0.1 nM Less than 0.2nM, less than 0.3nM, less than 0.4nM, less than 0.5nM, 0.6nM Less than 0.7nM, less than 0.8nM, less than 0.9nM, less than 1nM, less than 2nM, 3 Less than nM, less than 4nM, less than 5nM, less than 6nM, less than 7nM, less than 8nM, less than 9nM , less than 10 nM, less than 11 nM, less than 12 nM, less than 13 nM, less than 14 nM, 15 nM Less than 16nM, less than 17nM, less than 18nM, less than 19nM, less than 20nM, 21 Less than nM, less than 22nM, less than 23nM, less than 24nM, less than 25nM, less than 26nM, Less than 27 nM, less than 28 nM, less than 29 nM, less than 30 nM, less than 40 nM, or 5 Affinity (Kd or EC) to the target molecule (to which it specifically binds) less than 0 nM 50 ) has.

[0050] Molecules such as antibodies are able to identify specific cells, substances, or epitopes, and they are not able to identify other cells, substances, or epitopes. More frequently, more quickly, than reacting with or associating with cells, substances, or epitopes. If they react or associate with a longer duration and / or with higher affinity, It is said that antibodies exhibit "specific binding" or "preferential binding." Antibodies are those that bind to other substances or Rather than binding to an epitope, for example, by a statistically significant amount, with higher affinity and binding activity... If it binds more rapidly and / or for a longer duration, the target molecule or To "specifically bind" or "preferentially bind" to a pitope. Typically, specific binding. One member of a pair of molecules exhibiting a specific spatial relationship with the other member of that pair. and / or specifically bind to polar organization, and therefore are complementary to it. It has an area or cavity on its surface. Therefore, members of a pair are unique to each other. It possesses the property of binding. For example, it binds specifically or preferentially to specific epitopes. Antibodies bind to their specific epitopes more effectively than other epitopes. With higher affinity and binding activity, antibodies bind more rapidly and / or for a longer duration. This definition also applies to, for example, binding specifically or preferentially to a first target. The antibody (or partial or epitope) specifically or preferentially binds to a second target. It is understood by reading it as "it is fine, or it is not." The term is also, for example, In situations where antibodies are specific to certain epitopes held by certain antigens, It can also be applied to combinations, in which case a specific binding men possessing an antigen-binding fragment or domain. The bar can bind to various antigens that possess its epitope; for example, it Cross-reactivity with several different types of target antigens from multiple species that share a common epitope. It could be sexual.

[0051] Immunological binding generally refers to the binding of immunoglobulin molecules to specific immunoglobulin molecules. Non-covalent interactions that occur between an antigen and an electrostatic agent, for example, and not limited to, electrostatic agents. Attractive or repulsive forces, steric forces, hydrogen bonds, ionic, hydrophilic, and / or hydrophobic forces This refers to interactions resulting from van der Waals forces, as well as other interactions. The strength or affinity of epidemiological binding interactions can be expressed in terms of the interaction dissociation constant (Kd). This is possible, and in that case, a smaller Kd represents a higher affinity. The immunological binding properties of butylate are quantified using methods well known in this field. One such method involves the formation and dissociation of antigen-binding sites / antigen complexes. It is necessary to measure the rate, in which case those rates are the concentration of the complex partner. , the affinity of the interaction, and geometric parameters that equally affect the velocity in both directions It depends on the "on-velocity constant" (Kon) and the "off-velocity constant" (Koff). Both can be determined by calculating the concentration as well as the actual rates of association and dissociation. It follows. The Koff / Kon ratio allows for the release of all parameters not related to affinity and is thus equal to the dissociation constant Kd. As used herein, the term "affinity" includes the equilibrium constant for the reversible binding of two agents and is expressed as Kd or EC 50 as expressed for the ligand of a binding protein such as the affinity of an antibody for an epitope. The affinity can be, for example, from about 100 nanomolar concentration (nM) to about 0.1 nM, from about 100 nM to about 1 picomolar concentration (pM), or from about 100 nM to about 1 femtomolar concentration (fM). As used herein, the term "binding activity" refers to the resistance to dissociation after dilution of a complex of two or more agents. In some embodiments , the affinity is expressed with respect to the maximum half - effect concentration (EC 50 ), which refers to the concentration of an agent such as an antibody disclosed herein that induces a response at the midpoint between the baseline and the maximum value after a specific exposure time. EC is generally used as a measure of the potency of an antibody. 50 is generally used as a measure of the potency of an antibody. The antibodies can be prepared by any of a variety of techniques known to those of skill in the art. For example, see Harlow

[0052] and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. Monoclonal antibodies specific for the polypeptide of interest can be prepared, for example, using the techniques of Kohler and Milst ein, Eur. J. Immunol. 6:511 - 519, 1976 and modifications thereof. Methods of utilizing transgenic animals such as mice to express human antibodies are also available. It is also included. For example, Neuberger et al., Nature Biotechnology 14:826, 1996;Lonbe rg et al., Handbook of Experimental Pharmacology 113:49-101, 1994; and Lonberg See et al., Internal Review of Immunology 13:65-93, 1995. For specific cases, see REG. VELOCIMMUNE® platform by ENEREX® Examples include (see, for example, U.S. Patent No. 6,596,541).

[0053] Antibodies can also be used with phage display or yeast display libraries. It can be fabricated or identified (for example, U.S. Patent No. 7,244,592; Ch (See ao et al., Nature Protocols. 1:755-768, 2006). Unlimited availability of libraries. Qualitative examples include the structure of cloned or synthetic libraries, such as a human antibody repertoire. The diversity is represented by seven heavy chain variable region genes and seven light chain variable region genes. One example is the Human Combinatorial Antibody Library (HuCAL). These genes The combination of these children gives rise to 49 frameworks in the master library. Overlaying a highly variable gene cassette (CDR = Complementarity Determination Region) onto this framework. This allows for the reproduction of a vast repertoire of human antibodies. Human antibodies encoding the light chain variable region... Fragments derived from human donors, including heavy chain CDR-3, and heavy chain CDR-1. Synthetic DNA encoding diversity in, and heavy chain CDR-2 encoding diversity This also includes human libraries designed using synthetic DNA. Ibrary's role will be obvious to those skilled in the art.

[0054] In a particular embodiment, an antibody and its antigenicity as described herein Compatibility fragments provide support to the CDR and limit the spatial relationships of the CDRs relative to each other. Between the Chain Framework Region (FR) set and the Light Chain Framework Region (FR) set Includes heavy chain CDR sets and light chain CDR sets inserted into each. In this context, the term "CDR set" refers to the three hypervariable regions of the V region of the heavy or light chain. Starting from the N-terminus of the heavy or light chain, these regions are called "CDR1," respectively. These are labeled as "CDR2" and "CDR3". Therefore, the antigen-binding site is the heavy chain. Includes six CDRs, each containing a CDR set derived from the V region and the light chain V region, respectively. Polypeptides containing CDRs (e.g., CDR1, CDR2, or CDR3) are, In detail, it is called a "molecular recognition unit." Crystallographic analysis of several antigen-antibody complexes is C The amino acid residues of DR form extensive contact with the bound antigen, and the most extensive antigenic contact is It has been demonstrated that it is heavy chain CDR3. Therefore, its molecular recognition unit is mainly anti It is involved in the specificity of the proto-binding site.

[0055] As used herein, the term "FR set" refers to the C region of the heavy chain V region or the light chain V region. This refers to the four flanking amino acid sequences that form the framework of the CDR in the DR set. The residues can come into contact with the bound antigen; however, FR directs the V region to the antigen-binding site. FR is primarily involved in folding, and in particular, the FR residue is directly adjacent to the CDR. Within this molecule, certain amino residues and certain structural features are highly conserved. In this respect, all V region sequences are internal disulfide sequences of approximately 90 amino acid residues. It contains a fidoloop. When the V region is folded to the binding site, the CDR becomes an antigen. It is displayed as a protruding loop motif that forms a bonding surface. (Accurate CD) (Regardless of the R amino acid sequence) the CDR loop falls into a specific "canonical" structure. It is generally recognized that there are conserved structural regions of FR that influence the resulting shape. Furthermore, certain FR residues are non-covalent bonds that stabilize the interaction between antibody heavy and light chains. It is known to be involved in contact between sex domains.

[0056] The structure and location of the immunoglobulin variable domain are described by Kabat, EA et al., Sequences o f Proteins of Immunological Interest. 4th Edition. US Department of Health and H This can be determined by referring to uman Services. 1987 and its updates.

[0057] Monoclonal antibodies are involved in the selective binding of epitopes (naturally occurring, and "Monoclonal" refers to a homogeneous group of antibodies composed of amino acids (that do not exist in nature). "Antibodies are also included. Monoclonal antibodies are highly specific and have a single epitope." It is directed toward. The term "monoclonal antibody" refers to an intact monoclonal antibody. Not only whole and full-length monoclonal antibodies, but also their fragments (e.g., Fab, Fab', F(ab')2, Fv), single-stranded (ScFv), its variants, containing the antigen-binding moiety. Fusion proteins, humanized monoclonal antibodies, chimeric monoclonal antibodies, and necessary Antigen-binding fragments that possess specificity and the ability to bind to epitopes (epitope recognition) It also includes any other modified stereochemistry of immunoglobulin molecules, including the site of the antibody. Source or the method by which it is produced (e.g., hybridoma, phage selection, recombinant generation) The term is not intended to be limited to (currently, to transgenic animals). This includes immunoglobulins as a whole, plus the fragments described above under the definition of "antibodies." .

[0058] The proteolytic enzyme papain preferentially cleaves IgG molecules into several fragments. It is generated, and two of its fragments (F(ab) fragments) each contain intact antigen-binding sites. It forms a covalent heterodimer. The enzyme pepsin cleaves the IgG molecule, and A few fragments can be provided, and these fragments contain both antigen-binding sites F(ab ')2 fragments are given. The Fv fragment for use in a particular embodiment is IgM, Very rarely, due to preferential proteolytic cleavage of IgG or IgA immunoglobulin molecules Fv fragments can be generated. However, Fv fragments are more generally known in the art. It is derived using recombinant technology. The Fv fragment is involved in the antigen recognition and binding of the natural antibody molecule. A non-covalent VH::VL heterodiode containing antigen-binding sites that retain most of the binding ability. Includes Mar. Inbar et al., PNAS USA. 69:2659-2662, 1972; Hochman et al., Biochem. See 15:2706-2710, 1976; and Ehrlich et al., Biochem. 19:4091-4096, 1980.

[0059] In a particular embodiment, a single-chain Fv(scFv) antibody is intended. For example, Papa body (Ill et al., Prot. Eng. 10:949-57, 1997); Mini body (Martin et al., EMBO J 13:5305-9, 1994; Diabody (Holliger et al., PNAS 90: 6444-8, 1993) ); or Janusins ​​(Traunecker et al., EMBO J 10: 3655-59, 1991; and T Raunecker et al., Int. J. Cancer Suppl. 7:51-52, 1992) possess the desired specificity. Regarding the selection of antibodies, they were prepared using standard molecular biology techniques in accordance with the teachings of this application. It is possible.

[0060] Single-chain Fv(scFv) polypeptides are linked by a peptide-coding linker. Covalent genes expressed from a gene fusion containing the VH coding gene and VL coding gene It is a combined VH::VL heterodimer. Huston et al. (PNAS USA. 85(16):5879-5883) 1988). It folds into a three-dimensional structure that is substantially similar to the structure of the antigen-binding site, anti Naturally aggregated (but chemically separated) light chain polypeptides derived from the V region Several methods for identifying the chemical structure to convert heavy chain polypeptides into scFv molecules This is described in, for example, in Huston et al.'s U.S. Patent No. 5,091,513. and Patent No. 5,132,405; and Ladner et al.'s U.S. Patent No. 4,946, See specification No. 778.

[0061] In a particular embodiment, the antibody or its antigen-binding fragment is humanized. These embodiments involve antigen-binding sites derived from immunoglobulins from non-human species, as well as The remaining immunoglobulin structure of the molecule based on the structure and / or sequence of immunoglobulins This refers to a chimeric molecule that has the antigen-binding portion, generally prepared using recombinant technology. The position is either a complete variable domain fused onto a steady domain, or a suitable position within a variable domain. It may include only CDRs grafted onto a specific framework domain. The link-binding site may be in the wild type or modified by one or more amino acid substitutions. It can be altered. This removes the constant region as an immunogen in the human organism, but can be altered by foreign substances. The possibility of an immune response to the mutational region remains (LoBuglio et al., PNAS USA 86:4220-4224). , 1989;Queen et al., PNAS USA. 86:10029-10033, 1988;Riechmann et al., Nature. 332:323-327, 1988). Exemplary methods for the humanization of antibodies are listed in the U.S. Patent Publication No. 7,462. Examples include the methods described in Specification No. 697.

[0062] Another approach is not only to provide a human-derived constant region, but also to make it as close to human-like as possible. In order to reshape them accordingly, we will also focus on modifying the variable regions. . Relatively conserved in a given species, and presumably, in relation to the complementarity determination region (CDR) The heavy chain is flanked by four framework regions (FRs) that provide a scaffold. The variable regions of both the nucleotide and the light chain change in response to the epitope in question and determine the binding ability. It is known to contain the complementarity determining regions (CDRs). When a non-human antibody is prepared with respect to a particular epitope, its variable region can be "refashioned" or "humanized" and modified by grafting the CDRs derived from the non-human antibody onto the FRs present in human antibodies. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. In some embodiments, the humanized antibody preserves all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs derived from a mouse antibody). In other embodiments, the humanized antibody has one or more CDRs (one, two, three, four, five, six) that are changed with respect to the original antibody, and those CDRs are also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody. When prepared with respect to a particular epitope, its variable region can be "refashioned" or "humanized" and modified by grafting the CDRs derived from the non-human antibody onto the FRs present in human antibodies. When prepared with respect to a particular epitope, its variable region can be "refashioned" or "humanized" and modified by grafting the CDRs derived from the non-human antibody onto the FRs present in human antibodies. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. The application of this approach to various antibodies has been reported by Sato et al., Cancer Res. 53: 851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. In some embodiments, the humanized antibody preserves all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs derived from a mouse antibody). In some embodiments, the humanized antibody preserves all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs derived from a mouse antibody). In other embodiments, the humanized antibody has one or more CDRs (one, two, three, four, five, six) that are changed with respect to the original antibody, and those CDRs are also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody. In other embodiments, the humanized antibody has one or more CDRs (one, two, three, four, five, six) that are changed with respect to the original antibody, and those CDRs are also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody. In other embodiments, the humanized antibody has one or more CDRs (one, two, three, four, five, six) that are changed with respect to the original antibody, and those CDRs are also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody.

[0063] In certain embodiments, the antibody is a "chimeric" antibody. In this regard, chimeric Antibodies are operably linked to or otherwise fused with heterogeneous Fc moieties of different antibodies. It is composed of an antigen-binding fragment of an antibody. In a particular embodiment, the Fc domain and The heterogeneous Fc domain is of human origin. In certain embodiments, the Fc domain Alternatively, the heterologous Fc domain is of mouse origin. In other embodiments, the heterologous Fc domain The antibody may originate from a different Ig class than the parent antibody, and that class includes IgA (subclass IgA1 and IgA2, IgD, IgE, IgG (subclass IgG1, Ig Examples include G2, IgG3, and IgG4, and IgM. Further implementations In this state, the heterogeneous Fc domain is derived from one or more different Ig classes of CH2 And it may consist of a CH3 domain. As mentioned above regarding humanized antibodies. The antigen-binding fragment of the chimeric antibody is one or more of the CDRs of the antibodies described herein. Number only (for example, one, two, three, four, five, or six of the antibodies described herein) It may include one CDR, or the entire variable domain (VL, VH, or both). .

[0064] As used herein, subjects at risk of developing disease or adverse reactions are: If you have a detectable disease or symptoms of a disease, before taking any of the treatment methods described herein, Sometimes they are present, and sometimes they are not, and they show a detectable disease or symptoms of a disease. Sometimes it is indicated, and sometimes it is not. "There is a risk" is not stated in this specification. Measurable parameters that correlate with the onset of disease, as known in the art, This means that the subject has one or more risk factors. A subject having one or more of these risk factors is not considered to have one or more of these risk factors. It has a higher probability of developing disease or adverse reactions than the target population.

[0065] "Biocompatible" generally means that it does not harm the biological function of cells or objects, and is The material will not produce any unacceptable toxicity, including allergenicity and disease conditions. This refers to a compound.

[0066] The term "bonding" includes interactions between two molecules, such as salt bridges and water bridges. , covalent, electrostatic, hydrophobic, and ionic properties, and / or the interaction of hydrogen bonds This refers to a direct meeting conducted for practical purposes.

[0067] The term "bioavailability" refers to the amount of an active substance administered to a patient (for example, an anti- This refers to the whole-body availability of a substance (e.g., blood / plasma levels). Rity measures both the time (rate) and total amount (percentage) that reaches systemic circulation from the administered dosage form. This is an absolute term that indicates a constant value.

[0068] Through this disclosure, unless the context requires it to be interpreted in a different way, the word "comprise" is used. "Comprises" and "comprising" are the steps described. This means the inclusion of an element, a step, or a group of elements, but any other step or This is understood not to mean the exclusion of an element, a step, or a group of elements.

[0069] "Consisting of" means that what follows this phrase "consisting of" includes anything and is limited to those. Therefore, the phrase "consisting of" indicates that the listed elements are required or essential, and that no other elements can exist. "Consisting essentially of" means that it includes any element listed after this phrase, and is limited to other elements that do not interfere with or contribute to the activity or function specified in the disclosure for the listed elements. Therefore, the phrase "consisting essentially of" indicates that the listed elements are required or essential, but other elements are optional, and whether they exist depends on whether they significantly affect the activity or function of the listed elements, and they may or may not exist. Following this, it means including anything and being limited to those. Therefore, the phrase "consisting of" indicates that the listed elements are required or essential , and that no other elements can exist. "Consisting essentially of" means including any element listed after this phrase, and being limited to other elements that do not interfere with or contribute to the activity or function specified in the disclosure for the listed elements. Therefore, the phrase "consisting essentially of" indicates that the listed elements are required or essential, but other elements are optional , and whether they exist depends on whether they significantly affect the activity or function of the listed elements, and they may or may not exist. In the context of antibodies, the term "effector function" refers to the ability of the antibody to associate with other arms of the immune system, such as through activation of the classical complement pathway or through association with Fc receptors. The complement-dependent pathway is mainly driven by the interaction of C1q in the C1 complex with the clustered antibody Fc domain. Antibody-dependent cell-mediated cytotoxicity (ADCC) is mainly driven by the interaction of Fc receptors (FcRs) on the surface of effector cells (natural killer cells, macrophages, monocytes, and eosinophils) that bind to the Fc region of IgG that is already bound to the target cell. Fc receptors (FcRs) are involved in antibody-mediated

[0070] In the context of antibodies, the term "effector function" refers to the ability of the antibody to associate with other arms of the immune system, such as through activation of the classical complement pathway, or through association with Fc receptors. The complement-dependent pathway is mainly driven by the interaction of C1q in the C1 complex with the clustered antibody Fc domain. Antibody-dependent cell-mediated cytotoxicity (ADCC) is mainly driven by the interaction of Fc receptors (FcRs) on the surface of effector cells (natural killer cells, macrophages, monocytes, and eosinophils) that bind to the Fc region of IgG that is already bound to the target cell. Fc receptors (FcRs) are involved in antibody-mediated refers to the ability of the antibody to associate with other arms of the immune system, such as through activation of the classical complement pathway, or through association with Fc receptors. The complement-dependent pathway is mainly driven by the interaction of C1q in the C1 complex with the clustered antibody Fc domain. Antibody-dependent cell-mediated cytotoxicity (ADCC) is mainly driven by the interaction of Fc receptors (FcRs) on the surface of effector cells (natural killer cells, macrophages, monocytes, and eosinophils) that bind to the Fc region of IgG that is already bound to the target cell. Fc receptors (FcRs) are involved in antibody-mediated cytotoxicity (ADCC) is mainly driven by the interaction of Fc receptors (FcRs) on the surface of effector cells (natural killer cells, macrophages, monocytes, and eosinophils) that bind to the Fc region of IgG that is already bound to the target cell. Fc receptors (FcRs) are involved in antibody-mediated refers to the ability of the antibody to associate with other arms of the immune system, such as through activation of the classical complement pathway, or through association with Fc receptors. The complement-dependent pathway is mainly driven by the interaction of C1q in the C1 complex with the clustered antibody Fc domain. Antibody-dependent cell-mediated cytotoxicity (ADCC) is mainly driven by the interaction of Fc receptors (FcRs) on the surface of effector cells (natural killer cells, macrophages, monocytes, and eosinophils) that bind to the Fc region of IgG that is already bound to the target cell. Fc receptors (FcRs) are involved in antibody-mediated It is an important immune regulatory receptor that connects the (humoral) immune response to cellular effector function. Receptors for all classes of immunoglobulins have been identified, including FcγR(IgG) and F cεRI(IgE), FcαRI(IgA), FcμR(IgM), and FcδR(I Includes gD). At least three receptors for human IgG found on leukocytes. LAS: CD64 (FcγRI), CD32 (FcγRIIa, FcγRIIb, and F These include cγRIIc and CD16 (FcγRIIIa and FcγRIIIb). FcγRI is classified as a high-affinity receptor (nanomolar concentration range KD), while Fc γRII and FcγRIII have low to medium affinity (micromolar concentration range KD). Fc binding triggers a signaling pathway, which in turn facilitates the destruction of target cells. Various substances such as soluble enzymes, perforin, granzymes, and tumor necrosis factor are involved. It produces the secretion of quality. The level of ADCC effector function depends on the human IgG subtype. It differs. This depends on the allotype and specific FcvR, but simply put, A The DCC effector function is "high" for human IgG1 and IgG3, and IgG2 The levels of IgG4 and other factors are "low".

[0071] The terms "endotoxin-free" or "virtually endotoxin-free" refer to endotoxins. Even if the amount of xin is trace (for example, an amount that does not produce clinically harmful physiological effects in the subject), This refers to compositions, solvents, and / or containers containing undetectable amounts of endotoxin. This generally refers to endotoxins, which are produced by certain microorganisms, such as bacteria, typically. Although it is a toxin associated with Gram-negative bacteria, endotoxin is found in Listeria monocytoxin. It can be found in Gram-positive bacteria such as Listeria monocytogenes. It is the most commonly observed. Endotoxins are lipopolysaccharides (LPS) found in the outer membranes of various Gram-negative bacteria. It is lipooligosaccharide (LOS), which is the ability of these bacteria to cause disease. This exhibits the central pathogenic characteristics. Small amounts of endotoxin in humans are harmful in several ways. Among the physiological effects, fever, a decrease in blood pressure, and activation of inflammation and coagulation may occur. .

[0072] Therefore, in pharmaceutical manufacturing, endoto is released from the pharmaceutical product and / or drug container. In many cases, it is desirable to remove most or all traces of the toxin, even in small amounts, to humans. This is because it can cause harmful effects. For this purpose, depyrologie An oven can be used, and temperatures exceeding 300°C are necessary to destroy most endotoxins. This is because a certain degree is typically required. For example, the main syringe or vial. Based on the packaging material, the combination of a glass temperature of 250°C and a holding time of 30 minutes is This is often sufficient to achieve a 3-log reduction in endotoxin levels. Other methods for removing the substance have been devised, including, as described herein, and Examples of known methods in this field include chromatography and filtration. .

[0073] Endotoxins can be detected using routine techniques known in this field. It can. For example, Limulus Amoebocyte Lysate (Using blood derived from horseshoe crabs) is extremely useful for detecting the presence of endotoxins. This is a highly sensitive assay. In this test, very low levels of LPS are detected. The detectable coagulation of the lysate is amplified by a potent enzyme cascade that amplifies this reaction. Endotoxins can also be detected through enzyme-linked immunosorbent assay (ELISA). It can be quantified by ). Because it is virtually endotoxin-free, Toxin levels were less than approximately 0.001 EU per 1 mg of the active compound, and less than 0.005 EU. Full, less than 0.01 EU, less than 0.02 EU, less than 0.03 EU, less than 0.04 EU, 0. Less than 0.05 EU, less than 0.06 EU, less than 0.08 EU, less than 0.09 EU, less than 0.1 EU Full, less than 0.5 EU, less than 1.0 EU, less than 1.5 EU, less than 2 EU, less than 2.5 EU, Less than 3 EU, less than 4 EU, less than 5 EU, less than 6 EU, less than 7 EU, less than 8 EU, less than 9 EU It may be full or less than 10 EU. Typically, 1 ng of lipopolysaccharide (LPS) is about 1 Supports up to 10 EU.

[0074] An "epitope" is a binding pocket or paratope in the variable region of an antibody, and its binding interaction with that pocket. A binding phase that has the ability to form a binding phase, containing that portion of an antigen or other polymer. The interaction may manifest as intermolecular contact with one or more amino acid residues of the CDR. Epitopes can be linear peptide sequences or composed of discontinuous amino acid sequences. (i.e., "three-dimensional structure" or "discontinuous"). Therefore, epitopes are anti The primary structure, for example, of the VP1 polypeptide, can be continuous or discontinuous. Binding proteins can recognize one or more amino acid sequences; therefore Therefore, an epitope can define more than one different amino acid sequence. In a particular embodiment, The epitope is a reference sequence or a target molecule described herein (see, for example, Table V1). (Ref) Approximately or at least approximately 3, 4, 5, 6, 7, 8, 9, 10, 11 1, 12, 13, 14, 15, 16, 17, 18, 19, or 20 pieces, or approximately 3 or fewer, 4 or fewer, 5 or fewer, 6 or fewer, 7 or fewer, 8 or fewer, 9 or fewer , 10 or less, 11 or less, 12 or less, 13 or less, 14 or less, 15 or less, 16 The following are consecutive amino acids of 17 or fewer, 18 or fewer, 19 or fewer, or 20 or fewer ( That is, linear epitopes) or discontinuous amino acids (that is, structural epitopes) ) contains, consists of, or is essentially composed of. By binding proteins The epitopes that are recognized are peptide mapping and sequence analysis techniques well known to those skilled in the art. It can be determined by the procedure.

[0075] The term "maximum half-dose effective concentration" or "EC 50 " is the baseline and a specific exposure time A working substance, such as those disclosed herein, that induces an intermediate response with respect to the later maximum value. Refers to the concentration of (for example, an antibody); therefore, the EC of a stepwise dose-response curve. 50 is the most effective 50% of the values ​​represent the concentration of the compound observed. EC 50 Also, the maximum effect in vivo This represents the plasma concentration required to obtain 50%. For example, in some cases, EC 50 teeth This is the antibody concentration at which 50% of the viral infection is neutralized. Similarly, "EC 90 " is the most This refers to the concentration of the active substance or composition at which 90% of the major effect is observed. For example, in some cases , EC 90is the concentration of an antibody at which 90% of the viral infection is neutralized. "EC 90 " is , "EC 50 " and can be calculated from the Hill slope, or it can be determined directly from the data using ordinary knowledge in the art . In some embodiments, the EC of an agent (e.g., an antibody) is less than about 0.01 nM, 0.05 nM 50 less than, 0.1 nM less than, 0.2 nM less than, 0.3 nM less than, 0.4 nM less than, 0.5 nM less than, 0.6 nM less than, 0.7 nM less than, 0.8 nM less than, 0.9 nM less than, 1 nM less than, 2 nM less than, 3 nM less than, 4 nM less than, 5 nM less than, 6 nM less than, 7 nM less than, 8 nM less than, 9 nM less than, 10 nM less than, 11 nM less than, 12 nM less than, 13 nM less than, 14 nM less than, 15 nM less than, 16 nM less than, 17 nM less than, 18 nM less than, 19 nM less than, 20 nM less than, 25 nM less than, 30 nM less than, 40 nM less than, 50 nM less than, 60 nM less than, 70 nM less than, 80 nM less than, 90 nM less than, 100 nM less than, 200 nM less than, or 500 nM less than. In some embodiments, the agent has an EC value of about 1 nM or less. EC 50 value.

[0076] The term "maximum half-inhibitory concentration" or "IC 50 " refers to the concentration of an agent ( e.g., an antibody) described herein that induces a signal at the midpoint (50%) between the baseline control and the maximum possible signal . For example, in one case, IC 50 is the concentration of an antibody at which 50% of the available binding sites on the VP1 antigen are occupied.

[0077] The "half-life" of an active substance, such as an antibody, refers to the pharmacological, physiological, or other active effects of the active substance. Sex, in relation to such activity at the time of administration to the serum or tissue of an organism or It can refer to the time it takes to lose half of something relative to any other defined point in time. "Half-life" is Furthermore, the amount or concentration of the active substance at the time of administration to the serum or tissue of the organism For quantities or concentrations such as, or for any other specified time point, the blood of a living organism Half-life can refer to the time it takes for half of the initial dose administered to the tissue or body to decrease. The period can be measured in serum and / or any one or more selected tissues. can.

[0078] The terms "adjusting" and "changing" typically refer to statistically adjusting something relative to a control. To "increase" or "enhance" by a significant or physiologically significant amount or degree. or “to stimulate,” plus “to reduce” or “to decrease.” The "increased," "stimulated," or "enhanced" amounts are typically "statistically" A "significant" amount produced by the absence of the composition (e.g., the absence of the active substance) or by the control composition. 1.1 times, 1.2 times, 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times the amount produced x, 9x, 10x, 15x, 20x, 30x, 40x, 50x, 60x, 70x, 80x 90 times, 100 times, or more (for example, 500 times, 1000 times) (all in between) Includes integers and ranges. For example, increasing integers such as 1.5, 1.6, 1.7, 1.8, etc. It may include. The amount that "decreased" or "reduced" is typically a "statistically significant" amount. The amount produced by the composition, the absence of the composition (e.g., the absence of the active substance), or the control composition. 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 1 3%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 3 5%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 8 A decrease of 5%, 90%, 95%, or 100% (including all integers and ranges in between) This may include. Examples of comparative and "statistically significant" quantities are provided herein.

[0079] "Neutralization" is indicated by the absence of viral gene expression in the host cell, such as the absence of viral gene expression. It refers to the inhibition of infection. I don't intend to be bound by any one theory, but by specific antibodies The neutralization mechanism blocks the interaction of the viral capsid protein with its cell surface receptor. This involves the process of entry and transport of the viral genome before its delivery to the nucleus of the host cell. This could involve the destruction of the stage of intent.

[0080] The terms "polypeptide," "protein," and "peptide" are used interchangeably. This refers to a polymer of amino acids, not limited to any specific length. (Term: "enzyme") This includes catalysts for polypeptides or proteins. The terms include myristoylation and sulfation. This includes modifications such as glycosylation, phosphorylation, and addition or deletion of signal sequences. The term "polypeptide" or "protein" means one or more chains of amino acids. Each chain contains amino acids covalently linked by peptide bonds, and its polyp Peptides or proteins linked non-covalently and / or by peptide bonds It contains multiple chains that are covalently linked to each other, and is a natural protein, that is, naturally They exist and are specifically produced by non-recombinant cells or genetically modified or recombinant cells. It may have the sequence of the protein being produced, as well as the amino acid sequence of the natural protein. Deletion from or from the native sequence, addition thereto, and / or one or more amino acids It may include molecules having acid substitution. In a particular embodiment, the polypeptide is one or multiple recombinant DNA molecules (typically not found in cells otherwise) It is likely composed of heterogeneous polynucleotide sequences or combinations of polynucleotide sequences. It is a "recombinant" polypeptide produced by recombinant cells containing ( ).

[0081] As used herein, the term “isolated” polypeptide or protein refers to the subject matter. Proteins are (1) typically found together with at least some other proteins. (2) Not containing the protein, and not containing other proteins from the same source, for example, from the same species. (3) not essentially contained, (4) expressed by cells of different species, (5) that it is natural Naturally, at least about 50% of the accompanying polynucleotides, lipids, carbohydrates, or other materials (5) The isolated protein is naturally associated with The portion of the protein that is attached (by covalent or non-covalent interactions) (6) It is not associated with a polypeptide that is not naturally present, (covalent or non-covalent) (7) Not naturally present, being operably attached by a bound interaction This means that such isolated proteins are genomic DNA, cDNA, and mRNA. A, or other RNAs may be encoded by, or may be of synthetic origin (of may be of s It may be of synthetic origin, or any combination thereof. The isolated protein is then used for its purpose (treatment, diagnosis, prevention, research, or other uses). ) would interfere with the proteins or polypeptides found in its natural environment. It contains virtually no other impurities. The term "isolated antibody" refers to an antibody with different antigen specificity. It contains antibodies that are substantially free of other antibodies. However, it specifically binds to one antigen. The isolated antibodies may exhibit cross-reactivity with other antigens. Furthermore, the isolated antibodies Antibodies may not substantially contain other cellular materials and / or chemical substances.

[0082] The terms "polynucleotide" and "nucleic acid" refer to mRNA, RNA, cRNA, and cDNA. , and DNA. The term typically refers to a ribonucleotide of at least 10 base pairs in length. Otide or deoxyribonucleotide, or a modification of either type of nucleotide It refers to the polymer form of either a variant or a variant nucleotide. The term is used for the single-stranded form of DNA. This includes double-stranded DNA. The terms "isolated DNA" and "isolated polynucleotide" are also used. And "isolated nucleic acids" do not contain the whole genome DNA of a particular species, but rather the isolated portion. It refers to a child. Therefore, there is one isolated DNA segment that encodes a polypeptide. Or a DNA segment containing multiple coding sequences is further substantially that DNA segment The ment is isolated from or purified from the whole genomic DNA of the species from which the ment is obtained. This refers to the state of being.

[0083] In a particular embodiment, any predetermined active substance in the composition (e.g., an antibody) The "purity" of a certain composition can be defined. For example, a particular composition may be defined as an example and not limited to Without using biochemical and analytical chemistry to separate, identify, and quantify compounds, High-performance liquid chromatography (HFL) is a well-known type of column chromatography that is frequently used. When measured by HPLC, the protein content is based on protein or weight-to-weight. And, including all decimals and ranges in between, at least 80%, 85%, 90%, 9 1%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 10 It may contain active substances such as polypeptide active substances that are 0% pure.

[0084] The term "reference sequence" generally refers to a nucleic acid coding sequence that is used for comparison with another sequence. Alternatively, it refers to an amino acid sequence. This includes those listed by name, as well as those listed in tables and sequence lists. All polypeptides and polynucleotide compounds described herein, including those that have been modified. The columns are included as reference arrays.

[0085] Certain embodiments involve polypeptides (e.g., antibodies) described herein and The biologically active "variants" and "fragments" of polynucleotides that code for this Includes. A "variant" is a single variant of a reference polypeptide or polynucleotide. It contains multiple substitutions, additions, deletions, and / or insertions (for example, tables and sequence listings). (See reference). Variant polypeptides or polynucleotides are described herein. Reference arrays like these and at least approximately 50%, 55%, 60%, 65%, 70%, 75%, 80% %, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98 Having %, 99%, or more sequence identity, similarity, or homology, and having part Contains an amino acid or nucleotide sequence that substantially preserves the activity of the reference sequence. A sequence of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 pieces, 12 pieces, 13 pieces, 14 pieces, 15 pieces, 16 pieces, 17 pieces, 18 pieces, 19 pieces, 20 pieces, 30 pieces, 40 pieces, 50 pieces, 60 pieces, 70 pieces, 80 pieces, 90 pieces, 100 pieces, 110 pieces, 12 0, 130, 140, 150, or more amino acids or nucleotides The addition, deletion, insertion, or substitution of a sequence differs from the reference sequence and alters the activity of the reference sequence. This also includes substantially retained sequences. In certain embodiments, additions or deletions are , including addition and / or deletion of the C-terminus and / or N-terminus.

[0086] As used herein, the term “sequence identity” means, for example, “50% identical sequence identity.” The term "sequence" refers to a sequence based on one nucleotide at a time across a comparison window. It refers to the degree of identicality based on each amino acid. Therefore, "sequence identity" The "percentage" is two optimally aligned values ​​across the comparison window. The sequences are compared to identify identical nucleic acid bases (e.g., A, T, C, G, I) or identical amino acid residues. Base (for example, Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Ph) e, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cy Determine the number of positions where s and Met exist in both arrays, and generate the number of matching positions. The number of matching positions is the total number of positions in the comparison window (i.e., the number of matching positions in the comparison window). Divide by the dow size, and multiply the result by 100 to obtain the percentage of sequence identity. This can be calculated by determining the optimal alignment of the array for aligning the comparison window. Inment is an algorithm (Wisconsin Genetics Software). GAP, BESTFIT, FAS in e Package Release 7.0 TA, and TFASTA, Genetics Computer Group, 575 On computers at Science Drive Madison, Wis., USA This may be done by execution or scrutiny, and the best alignment (i.e., the result) The one that produces the highest percentage homology across the comparison window is selected. It can be prepared by any of the following methods. For example, Altschul et al., Nucl. Acids Res. 25 As disclosed in :3389, 1997, refer to the BLAST family of programs as well. It is possible.

[0087] The term "solubility" refers to the degree of solubility of an active substance (e.g., an antibody) in a liquid solvent. This refers to the property of dissolving and forming a homogeneous solution. Solubility is typically measured per unit volume of solvent. Mass of solute per unit (g of solute per 1 kg of solvent, g of solute per 1 dL (100 mL), mg) (e.g., / ml), volume molar concentration, weight molar concentration, mole fraction, or other similar descriptions of concentration. It is expressed as a concentration by one of the following: the amount of solute that can dissolve per unit volume of solvent. The maximum equilibrium amount is determined under specific conditions, including temperature, pressure, pH, and solvent properties. This is the solubility of a solute in a solvent. In certain embodiments, solubility is physiological p H or other pH levels, for example, pH 5.0, pH 6.0, pH 7.0, pH 7.4, pH 7 It is measured at 0.6, pH 7.8, or pH 8.0 (for example, around pH 5-8). In the embodiment, the solubility is determined by water, or PBS or NaCl (containing NaPO4). It is measured in a physiological buffer such as (which does not contain). In a specific embodiment, The degree of solution is at a relatively low pH (e.g., pH 6.0) and a relatively high salt content (e.g., 5 Measurements are taken using 00 mM NaCl and 10 mM NaPO4. In certain embodiments In this context, solubility is measured in a biological fluid (solvent) such as blood or serum. In certain embodiments, the temperature is approximately room temperature (for example, approximately 20°C, 21°C, 22°C, 23°C). It may be 24°C, 25°C, or about body temperature (37°C). In a particular embodiment, The active substance is present at room temperature or 37°C at a concentration of at least approximately 0.1 mg / ml, 0.2 mg / ml, 0.3mg / ml, 0.4mg / ml, 0.5mg / ml, 0.6mg / ml, 0 .7mg / ml, 0.8mg / ml, 0.9mg / ml, 1mg / ml, 2mg / ml, 3mg / ml, 4mg / ml, 5mg / ml, 6mg / ml, 7mg / ml, 8mg / m l, 9mg / ml, 10mg / ml, 11mg / ml, 12mg / ml, 13mg / ml , 14mg / ml, 15mg / ml, 16mg / ml, 17mg / ml, 18mg / ml , 19mg / ml, 20mg / ml, 25mg / ml, 30mg / ml, 40mg / ml , 50mg / ml, 60mg / ml, 70mg / ml, 80mg / ml, 90mg / ml It has a solubility of 100 mg / ml.

[0088] "Target" or "Target that needs it" or "Patient" or "Patient that needs it" "Subject" includes mammals such as humans.

[0089] "Substantially" or "essentially" means almost entirely or completely, for example and This means 95%, 96%, 97%, 98%, 99%, or more of a given amount. do.

[0090] "Statistically significant" means that the result is unlikely to have occurred by chance. It tastes good. Statistical significance can be determined by any method known in the art. Yes, it is possible. A commonly used measure of significance is the p-value, which indicates whether the null hypothesis is true. In fact, it is the frequency or probability that the observed event will occur. The obtained p-value is significant. If the value is less than the threshold, the null hypothesis is rejected. In the simple case, the significance level is 0.05. It is defined by the following p-value.

[0091] "Therapeutic response" refers to the improvement of symptoms based on the administration of one or more therapeutic agents (whether the improvement is sustained or not). (Regardless of whether it is a fly or not)

[0092] As used herein, the terms “therapeutic effective dose,” “therapeutic dose,” and “prophylactic effective dose” are used. The "diagnostic effective dose," or "diagnostic effective dose," is the dose required to induce the desired biological response after administration. This refers to the amount of the active substance (for example, anti-polyomavirus antibodies).

[0093] When used herein, "treatment" refers to an object (e.g., a mammal such as a human) or a cell. "An intervention is any type of intervention used to alter the natural course of an individual or cell." Treatment may include, but is not limited to, the administration of a pharmaceutical composition, and the treatment may be preventive or ill. It may be implemented either at the onset of a specific event or after contact with a pathogen. “Prophylactic” This also includes treatment, which slows the rate of progression of the disease or condition that is being treated. To cause a bowel movement, to delay the onset of the disease or condition, or to reduce the severity of its onset. The aim may be to reduce the risk. "Treatment" or "prevention" does not necessarily mean the disease or This does not necessarily indicate complete eradication, cure, or prevention of the condition or its associated symptoms.

[0094] The term "wild type" refers to the gene or gene product that is most frequently observed in a population. Therefore, it is not arbitrarily designed to be the "normal" or "wild-type" form of that gene. ru.

[0095] Each embodiment in this specification is described in accordance with all other descriptions unless otherwise explicitly stated. This can be applied to the embodiments.

[0096] As described above, embodiments of the present disclosure include one or more polyomawi This relates to the administration regimen and optimal dosage of Russ neutralizing antibody, as well as its antigen-binding fragment.

[0097] For example, a particular embodiment involves BK or JC porting to a human subject that requires it. A regimen for the treatment of riomavirus infection, (a) Polyomaviruses that specifically bind to the VP1 protein A step of administering a target dose of the antibody or its antigen-binding fragment parenterally; (b) Measure the serum or tissue levels of the antibody or its antigen-binding fragment in the subject. The steps; and (c) Trough concentration in serum or tissue (C trough ) is approximately 3-860 μg / mL or less The step of administering an additional dose of the antibody or its antigen-binding fragment before the descent. The administration regimen includes, during the treatment, the antibody or its antigen-binding fragment in the serum or group The concentration in the fabric is C trough Regarding the administration regimen for maintaining a higher level.

[0098] "Trough concentration" or "C trough "This refers to the most active substance before administering the next dose. This refers to low serum or tissue concentrations. For regular administration, C trough Typically, For example, in order to maintain optimal virus neutralizing activity and avoid overdose, the following dosage The dose is measured immediately before administration.

[0099] In a particular embodiment, step (c) C trough Regarding plasma The administration regimen was determined, and the plasma C trough Approximately 150-860 μg / mL , or approximately 150 μg / mL, 160 μg / mL, 170 μg / mL, 180 μg / mL , 190μg / mL, 200μg / mL, 210μg / mL, 220μg / mL, 230 μg / mL, 240μg / mL, 250μg / mL, 260μg / mL, 270μg / m L, 280μg / mL, 290μg / mL, 300μg / mL, 310μg / mL, 32 0μg / mL, 330μg / mL, 340μg / mL, 350μg / mL, 360μg / mL, 370μg / mL, 380μg / mL, 390μg / mL, 400μg / mL, 4 10μg / mL, 420μg / mL, 430μg / mL, 440μg / mL, 450μg / mL, 460μg / mL, 470μg / mL, 480μg / mL, 490μg / mL, 500μg / mL, 510μg / mL, 520μg / mL, 530μg / mL, 540μ g / mL, 550μg / mL, 560μg / mL, 570μg / mL, 580μg / mL , 590μg / mL, 600μg / mL, 610μg / mL, 20μg / mL, 630μ g / mL, 640μg / mL, 650μg / mL, 660μg / mL, 670μg / mL , 680μg / mL, 690μg / mL, 600μg / mL, 710μg / mL, 720 μg / mL, 730μg / mL, 740μg / mL, 750μg / mL, 760μg / m L, 770μg / mL, 780μg / mL, 790μg / mL, 800μg / mL, 81 0μg / mL, 820μg / mL, 830μg / mL, 840μg / mL, 850μg / Before reducing the dose to less than mL or 860 μg / mL, administer further doses. include.

[0100] In some embodiments, step (c) C trough is a specific organization For example, this concerns kidney tissue or bladder tissue. For illustrative purposes only, antibodies Alternatively, the plasma concentration of the antigen-binding fragment is the same as the concentration in renal tissue (e.g., kidney C trough is plasma C trough (This is approximately 13.7% of the total) or concentration in bladder tissue (e.g., bladder C t rough is plasma C trough It can be used to determine (which is about 1%) ru.

[0101] In some embodiments, step (c) C trough It is in the kidney tissue The administration regimen is determined based on the amount of C in the renal tissue. trough Approximately 23.5-120μ g / mL, or approximately 23.5 μg / mL, 25 μg / mL, 30 μg / mL, 35 μg / mL, 40μg / mL, 45μg / mL, 50μg / mL, 55μg / mL, 60μg / mL, 65μg / mL, 70μg / mL, 75μg / mL, 80μg / mL, 85μg / mL, 90μg / mL, 95μg / mL, 100μg / mL, 105μg / mL, 110 Further administration before dropping to μg / mL, 115 μg / mL, or less than 120 μg / mL This includes administering a drug dose. In some of these and other related embodiments, The target population includes those who have or are at risk of developing BK virus-associated nephropathy.

[0102] In some embodiments, step (c) C trough It is in the bladder tissue The administration regimen was determined to be C in bladder tissue. trough Approximately 3-10 μg / mL, or approximately 3 μg / mL, 3.5 μg / mL, 4 μg / mL, 4.5 μg / mL, 5 μg / mL, 5.5 μg / mL, 6 μg / mL, 6.5 μg / mL, 7 μg / mL, 7. 5μg / mL, 8μg / mL, 8.5μg / mL, 9μg / mL, 9.5μg / mL, This includes administering additional doses before the concentration drops to less than 10 μg / mL. In some of the related embodiments, including the one mentioned above, the subject is BK virus-associated hemorrhagic bladder It has or is at risk of having flames.

[0103] This may be done either separately or as part of the dosing regimen described herein. Some embodiments include one or more polyomavirus neutralizing antibodies, in addition to This relates to the optimal dosage of the antigen-binding fragment. For example, in certain embodiments, it is necessary Methods for treating BK or JC polyomavirus infection in human subjects Therefore, polyomaviruses that specifically bind to the VP1 protein of polyomaviruses The step includes administering a neutralizing antibody or its antigen-binding fragment parenterally in a controlled dose. The dosage is approximately 10-30 mg / kg, for example, approximately 10 mg / kg, 11 mg / kg, 12mg / kg, 13mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17mg / kg, 18mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22mg / kg, 23mg / kg, 24mg / kg, 25mg / kg, 26mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, or 30 mg / kg, or approximately 10~15mg / kg, 10~20mg / kg, 10~25mg / kg, 15~20mg / kg, 15-25mg / kg, 15-30mg / kg, 20-30mg / kg, or The dosage is 25-30 mg / kg, according to the method.

[0104] The term "BKV" or "BK virus" refers to the Polyomaviridae family. This refers to members of the genus Orthopolyomavirus. Illus has a genome of approximately 5,000 base pairs, and is an icosahedron without an envelope. These are true-stranded DNA viruses. They have a diameter of approximately 40-45 nM (Bennett et al.). al., Microbes and Infection. 14:672-683, 2012). "JCV" or "JC virus" " is a member of the Polyomaviridae family, genus Orthopolyomavirus. This refers to a member of the olyomavirus. JCV is closely related to BKV, and similarly, it is approximately 5,0 It is an icosahedral, non-enveloped, double-stranded DNA virus with a genome of 00 base pairs. They have a diameter of approximately 40-45 nM (Johne et al., Arch. Virol. 156:1). 627-1634, 2011).

[0105] As mentioned above, antibodies or their antigen-binding fragments are polyomaviruses It specifically binds to the VP1 protein. The term "VP1" refers to a major polyomavirus. This refers to the capsid subunit protein. The "VP1 pentamer" is composed of five monomers of VP1. It is composed of the following. An example of the VP1 protein is provided in Table V1 below.

[0106] [Table 1]

[0107] In a particular embodiment, an antibody or its antigen-binding fragment is selected from Table V1. It specifically binds to one or more VP1 proteins. In certain embodiments, it inhibits The body or its antigen-binding fragments are each of the VP1 proteins shown in SEQ ID NOs: 1-5 It specifically binds to or cross-reacts with BKV serotypes I-IV. It binds to VP1 derived from all of the JC virus. In certain embodiments, the antibody also Its antigen-binding fragment neutralizes all BKV serotypes I-IV and JC virus, Alternatively, it reduces or inhibits its replication. In some embodiments, The antibody or its antigen-binding fragment is limited to one, two, or three fragments in VP1. Contact residues, for example, any one of Y169, R170, and K172 of VP1 or It specifically binds to VP1 structural epitopes that include multiple components.

[0108] In some embodiments, the antibody or its antigen-binding fragment is a 1 or Complementary determination region (CDR) sequence, for example, V H CDR1, V H CDR2, and V H CDR Heavy chain variable (V) containing 3 sequences H ) region, and V L CDR1, V L CDR2, and V L Light chain variable (V) containing one or more CDR sequences such as the CDR3 sequence L ) Depending on the region Characterized or including them. Exemplary V H , V H CDR1, V H CDR2, V H C DR3, V L , V L CDR1, V L CDR2, and V L The CDR3 sequence is shown in Table A1 below. And provided in Table A2.

[0109] [Table 2]

[0110] [Table 3]

[0111] Therefore, in a particular embodiment, the antibody or its antigen-binding fragment is, Table A1 V selected from H CDR1, V H CDR2, and V H V containing the CDR3 sequence H region, Furthermore, the polyomavirus VP1 protein (e.g., selected from Table V1) is specific to... The variants that join together; and the V selected from Table A1 L CDR1, V L CD R2 and V L V containing the CDR3 sequence L The region, and polyomavirus VP1 tan It includes variants that specifically bind to a protein (e.g., selected from Table V1). In a particular embodiment, the CDR sequence is as follows: V H CDR1, V H C DR2, and V H The CDR3 sequence contains sequence numbers 6-8, respectively, V L CDR1, V L CDR2, and V L The CDR3 sequences each contain sequence numbers 9-11, and polyoma Another example is its variant that specifically binds to the viral VP1 protein. (CDR region) One or more of the following, for example, V as described herein H CDR1, V H CDR2, V H C DR3, V L CDR1, V L CDR2, and / or V L One or multiple CDR3 sequences Variants that have variations of one, two, three, four, five, or all six in number. This includes. Exemplary “changes” include amino acid substitutions, additions, and deletions.

[0112] In a particular embodiment, V H The arrays are selected from Table A2 and at least 8 They are 0%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical. For example, V HThe array contains one, two, or three elements in one or more framework regions. Examples include having four, five, or six variations. In some embodiments, , V L The sequences are selected from Table A2 and are at least 80%, 85%, 90%, and 95%. , 97%, 98%, 99%, or 100% identical, for example, V L The array is one This involves one, two, three, four, five, or six changes across multiple framework areas. Examples include cases where it has [this feature]. Specific examples include V5Q, G9P, T10G, N30S, N3 One or more substitutions selected from 0K and N30Q are listed. In the application form, the V of the antibody or antigen-binding fragment H and V L The array is as follows: V H The sequence contains, consists of, or is essentially derived from sequence number 12, V L The sequence contains, consists of, or is essentially derived from sequence number 13. In a typical embodiment, the antibody is MAU868 (P8D11 is similar).

[0113] Some administration regimens or methods are used for subjects with polyomavirus infection, for example The condition is to identify BK or JC viruria or BK or JC viremia. This includes making a diagnosis. In some embodiments, the administration regimen is tailored to the subject. This includes the step of identifying or diagnosing the BK virus genotype or JC virus. In some embodiments, polyomavirus infection is genotype I, II, III, The BK virus comprises one or more selected from and IV. In some embodiments, Polyomavirus infection is caused by BK virus genotype I, II, III, or IV. It includes only one of the following. In some embodiments, polyomavirus infection is BK virus Includes two, three, or all four of Rus genotypes I, II, III, and / or IV. In some embodiments, polyomavirus infection is caused by the JC virus alone. or any one of BK virus genotypes I, II, III, and / or IV, 2 Includes, in combination with three or four. Viremia, viruria, and B Methods for identifying, diagnosing, or measuring K virus genotypes and JC viruses are provided by this technology. It is known in the field of technology, and includes real-time PCR and high-resolution thaw analysis (HR MA), and other techniques (e.g., Luo et al., J Virol. 83:2285-2297, 2009;Gambarino et al., Mol Biotechnol. 49:151-8, 2011;Matsuda et al., J Med Vi rol. 83:2128-34, 2011; and Toan et al., Transplantation Proceedings. 51:2683-2 (See 688, 2019).

[0114] In some embodiments, subjects have significantly reduced immune system capacity to fight infectious diseases. "Immunocompromised" or "immunodeficiency" refers to a state of diarrhea or complete absence of diarrhea. It has. In some embodiments, immunodeficiency is secondary or acquired, and surgery, Damage, or various active substances, such as immunosuppressants related to organ or cell transplantation, Treatment with lucocorticoids, chemotherapeutic agents, and disease-modifying antirheumatic drugs, or heavy metals This is the result of exposure to environmental toxins such as pesticides or petrochemicals. Treatment-related aspects Regarding the term "immunosuppression," more generally, it refers to the beneficial effects of reducing the function of the immune system. This refers to beneficial and potentially harmful effects, while the term "immunodeficiency" primarily refers to the risk of infection. It refers to the adverse effects of increased immunodeficiency. In some cases, secondary immunodeficiency is a condition associated with a specific disease or state. It is more likely to be caused by: many types of cancer, especially cancers of the bone marrow and blood cells (for example, Examples include leukemia, lymphoma, multiple myeloma, and certain chronic infections. All is also acquired immunodeficiency syndrome, caused by the human immunodeficiency virus (HIV). This is also a characteristic of the group (AIDS). Various hormonal and metabolic disorders also contribute to immunodeficiency. These can cause anemia, hypothyroidism, and hyperglycemia. In some cases, immunodeficiency is associated with aging, for example, when the subject is around or at least around 60 or 65 years old. This applies if you are 70, 75, 80, 85, 90, 95, or 100 years old. In some cases, immunodeficiency is primary, resulting from a genetic disorder in the subject. It is congenital.

[0115] Examples of immunodeficiency include humoral immunodeficiency (including B-cell deficiency or dysfunction), Generally, this is due to hypogammaglobulinemia (a decrease in one or more types of antibodies) and and / or agammaglobulinemia (absence of all or most antibody production); for example T cell deficiency, characterized by decreased T cell count or delayed-type hypersensitivity skin test; A decrease in the number of granulocytes, such as a decrease in the number of neutral granulocytes (called neutropenia) (granulocytopenia) , or if absent, agranulocytosis) and granulocytes including decreased function of individual granulocytes Deficiency; asplenia characterized by a lack of splenic function; and impaired complement system function Characterized by complement deficiency.

[0116] In some embodiments, the subject is a transplantation procedure, e.g., organ transplantation or cell-based You are about to undergo, are undergoing, or have undergone the transplant procedure. The question is whether or not there are organ transplants. Examples of organ transplants include kidneys (or renal organs), and hearts. Examples include liver, lung, pancreas, intestine, thymus, and uterine transplants. In particular, hematopoietic stem cell transplantation (H) includes syngeneic, autologous, and allogeneic hematopoietic stem cell transplantation (HCT). CT) is one example. In certain embodiments, the transplantation is the transplantation of an allogeneic graft, i.e., The transplantation of organs, tissues, or cells between two genetically dissimilar members of the same species. In some embodiments, the subject is related to immunosuppressive therapy associated with the transplantation procedure. Or they have immunodeficiency caused by it.

[0117] In some embodiments, the target is one or more BK or JC virus infections. The patient exhibits the following symptoms. For example, in some cases, the subject may experience blurred vision or other visual changes, as well as brown discoloration. Other symptoms include red urine, painful urination, decreased kidney function, difficulty urinating, cough, common cold, shortness of breath, fever, muscle pain, Having one or more of the following: muscle weakness and / or seizures. Such symptoms include: For example, it can result from ureteral stricture, interstitial nephritis, or more commonly, kidney inflammation.

[0118] In certain embodiments, the subjects are BK virus-associated nephropathy and BK virus-associated hemorrhagic bladder. The patient has a condition selected from cystitis and JC virus-associated progressive multifocal leukoencephalopathy (PML). There is a risk of having or having the disease. The term "BKV nephropathy" or "BK virus-associated nephropathy" "BKVAN" is a viral cytopathic change and virus primarily occurring in the renal tubular epithelium. Inflammatory interstitial infection caused by lytic infection of BK virus, characterized by gene expression It refers to nephropathy. "Hemorrhagic cystitis" or "BK virus-associated hemorrhagic cystitis" is typically This refers to inflammation of the bladder, limited to lower urinary tract symptoms including dysuria, hematuria, and bleeding. "JC virus-associated PML" often causes progressive damage to the white matter of the brain in multiple locations (multifocal). This refers to a rare, often fatal, viral disease characterized by sores or inflammation.

[0119] As stated above, this specification includes methods for treating polyomavirus infections. The administration regimen described in the book is, for example, administered in serum or tissue C trough The most In order to achieve a dosing regimen that maintains a minimum level or minimum range, in the subject, The serum or tissue levels of polyomavirus neutralizing antibodies or their antigen-binding fragments are measured. The steps include monitoring or measuring. In some embodiments, (a) the dosage The time between the given step and the step to monitor or measure in (b) is approximately or less Each lasts approximately 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 1 0 days, 11 days, 12 days, 13 days, 14 days, 14 days, 16 days, 17 days, 1 8 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 2 6 days, 27 days, 28 days, 29 days, or 30 days, or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, or 1 month, 2 months Between, or 3 months, or about 1 day or less, 2 days or less, 3 days or less, 4 days or less, 5 Less than 10 days, 6 days or less, 7 days or less, 8 days or less, 9 days or less, 10 days or less, 11 days Below, 12 days or less, 13 days or less, 14 days or less, 14 days or less, 16 days or less, 17 Less than 18 days, 19 days or less, 20 days or less, 21 days or less, 22 days or less, 23 days or less, 24 days or less, 25 days or less, 26 days or less, 27 days or less, 28 days or less Below, 29 days or less, or 30 days or less, or 1 week or less, 2 weeks or less, or 3 weeks or more Below, 4 weeks or less, 5 weeks or less, 6 weeks or less, 7 weeks or less, or 8 weeks or less, It is one month or less, two months or less, or three months or less. Some embodiments are ( b) Serum or tissue levels in the subjects were monitored for approximately 3, 4, 5, 6, 7, and 8 days. , 9th, 10th, 11th, 12th, 13th, 14th, 14th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, Once every 29 or 30 days, or approximately every 1, 2, 3, 4, 5, or 6 weeks. Once a week, once every 7 or 8 weeks, three times a week, or approximately every 1, 2 or 3 months This includes a step of taking measurements 1 to 6 times.

[0120] In certain embodiments, with respect to polyomaviruses in a subject, an antibody or its anti- EC in serum or tissue of primordial-bound fragments 50 (relative to the than the serum) orthuk EC 50 ), serum or tissue C below the specified level trough It descends The procedure includes the step of administering a further dose of the antibody or its antigen-binding fragment. Furthermore, some of the administration regimens and methods provided herein are for the treatment of polyomavirus EC 50 C at the minimum level or within the range trough Maintain.

[0121] For example, in some embodiments, polyomavirus infection is caused by the BK virus gene Includes type I, C trough The tissue concentration in which, for example, the concentration in renal tissue, is the antibody or The antigen-binding fragment EC 50 From approximately 2618 times to approximately 3775 times, or approximately 13,0 A range up to 61 times (for example, approximately 2600 times, 3000 times, 3500 times, 4000 times, 45 00x, 5000x, 4500x, 5000x, 5500x, 6000x, 6500x, 7000x, 7500x, 8000x, 8500x, 9000x, 9500x, 10,0 00x, 10,500x, 11,000x, 11,500x, 12,000x, 12,5 (00 times, or 13,000 times), EC 50 Approximately 0.009 ± 0.010 μg / m² L. In some embodiments, polyomavirus infection is caused by the BK virus gene. Includes type I, C trough The tissue concentration in the region, for example, the concentration in bladder tissue, is related to the antibody and The EC of its antigen-binding fragment 50 Approximately 500 to 692 times, or approximately 1000 times Up to (for example, approximately 500 times, 600 times, 700 times, 800 times, 900 times, or 1 (000 times), EC 50 The concentration is approximately 0.009 ± 0.010 μg / mL.

[0122] In a particular embodiment, polyomavirus infection is associated with BK virus genotype II. Including, C troughThe tissue concentration in which, for example, the concentration in renal tissue, is the antibody or its anti- EC of primordial binding fragments 50 The range is approximately 589 times to approximately 849 times, or up to approximately 2942 times. (For example, approximately 600 times, 700 times, 800 times, 900 times, 1000 times, 1500 times, 2 (000 times, 2500 times, 3000 times), EC 50 Approximately 0.040 ± 0.025 μg It is / mL. In some embodiments, polyomavirus infection is BK virus gene Includes gene type II, C trough The tissue concentration in the region, for example, the bladder tissue concentration, is anti EC of the body or its antigen-binding fragments 50 From about 100 times to about 156 times, or about 20 Range up to 0x (for example, approximately 110x, 120x, 130x, 140x, 150x, 16 (0x, 170x, 180x, 190x, or 200x), EC 50 approximately 0.04 The concentration is 0 ± 0.025 μg / mL.

[0123] In some embodiments, polyomavirus infection is BK virus genotype III Includes C trough The tissue concentration in which, for example, the concentration in renal tissue, is the antibody or EC of antigen-binding fragments 50 From approximately 253 times to approximately 365 times, or up to approximately 1265 times Range (for example, approximately 250x, 300x, 350x, 400x, 450x, 500x, 55 0x, 600x, 650x, 700x, 750x, 800x, 850x, 900x, 95 (0x, 1000x, 1150x, 1200x, 1250x, 1300x) and EC5 0 is approximately 0.093 ± 0.057 μg / mL. In some embodiments, polio BK virus infection includes BK virus genotype III, C troughIn the organization The concentration, for example, the concentration in bladder tissue, is the EC of the antibody or its antigen-binding fragment. 50 Approximately 50 times From approximately 67 times, or up to approximately 100 times (for example, approximately 50 times, 55 times, 60 times, (65 times, 70 times, 75 times, 80 times, 85 times, 90 times, 95 times, or 100 times) EC 50 The concentration is approximately 0.093 ± 0.057 μg / mL.

[0124] In some cases, polyomavirus infection includes BK virus genotype IV, C tro ugh The tissue concentration in which, for example, the concentration in renal tissue, is the concentration of the antibody or its antigen-binding fragment. EC 50 A range of approximately 1122 times to approximately 1618 times, or up to approximately 5604 times (for example) , approximately 1100x, 1200x, 1500x, 2000x, 2500x, 3000x, 35 (00x, 4000x, 4500x, 5000x, 5500x, 6000x), EC 50 The concentration is approximately 0.021 ± 0.020 μg / mL. In some cases, polyomavirus The infection includes BK virus genotype IV, C trough Concentration in tissues, for example, Bladder tissue concentration is determined by the EC of the antibody or its antigen-binding fragment. 50 Approximately 100 times to approximately 297 Up to double, or in the range of approximately 500 times (for example, approximately 100 times, 150 times, 200 times, 25 (0x, 300x, 350x, 400x, or 500x), EC 50 approximately 0.02 The concentration is 1 ± 0.020 μg / mL.

[0125] In some cases, polyomavirus infections include JC virus, C trough ni oke The tissue concentration is the EC of the antibody or its antigen-binding fragment. 50 At least about 29 times to about Up to 110 times, up to approximately 158 times, up to approximately 547 times (for example, 20 times, 30 times, 40 times, 5 0x, 60x, 70x, 80x, 90x, 100x, 110x, 120x, 130x, 1 40x, 150x, 200x, 250x, 300x, 350x, 400x, 450x, 5 (00x, 550x, 600x) and EC 50 This is approximately 0.215 ± 0.130 μg / mL That is the case.

[0126] EC of polyomavirus neutralizing antibodies against BK virus genotype or JC virus 50 and C trough The level will be determined by the everyday techniques in this field. This is possible. For example, C trough Among several techniques, the level is particularly colorimetric. Measurement should be performed using a verified sandwich ELISA-based assay that employs detection. E-commerce is possible. 50 This can be measured in cell-based assays in ex vivo environments. (See Example 2 for an example).

[0127] Regarding in vivo use, as stated above, for the treatment of human diseases, The antibodies, their antigen-binding fragments, and other active ingredients described in the document are generally used before administration. It is incorporated into one or more pharmaceutical or therapeutic compositions.

[0128] Therefore, certain embodiments are as described herein, polyomau A pharmaceutically or This relates to therapeutic compositions. In some cases, the pharmaceutically or therapeutic compositions described herein. The substance contains polyomavirus neutralizing antibodies or their antigen-binding fragments, either pharmaceutically or physiologically. In combination with an acceptable carrier or excipient, it includes. In a particular embodiment, a carrier This includes histidine and / or sugars such as glycine and sucrose, as well as / or poly It contains polyols such as sorbates. In a specific embodiment, the support is histidine, Contains sucrose and polysorbate.

[0129] In some embodiments, the pharmaceutical compositions described herein are in the art. As is known, it does not significantly form aggregates, has the desired solubility, and It has an immunogenicity profile suitable for use in humans and / or in humans. Therefore, In some embodiments, a polyomavirus neutralizing antibody or its antigen-binding fragment is used. The composition, when measured by, for example, dynamic light scattering, is approximately 50%, 45%, 40%, and 3%. 5%, 30%, 25%, 20%, 15%, 14%, 13%, 12%, 11%, 10%, 9 %, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%, or less than approximately 50%, Less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15% Full, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8% Full, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% It has aggregates. Some compositions have non-aggregated antibodies or their antigen-binding fragments. Regarding the apparent molecular weight, at least about 50%, about 60%, about 70%, about 80%, and about 90% Polyomavirus neutralizing antibodies or their antigen-binding cleavage are monodispersible, or approximately 95% monodisperse. Includes pieces.

[0130] The composition is administered by various different routes, including parenteral and enteral administration. It can be obtained. Examples of parenteral administration include intravenous, subcutaneous, subarachnoid, epidural, intracerebral, ventricular, and nasal cavity. Administration methods include intramuscular, intra-arterial, and inhalation. Examples of enteral administration include oral or direct injection. Intestinal administration is one example. In certain embodiments, for example, an infusion pump or portable infusion device is used. or by IV injection, for example by intravenous bolus injection, by intravenous injection (for example, Approximately 10 to 90 minutes, or about 10, 20, 30, 40, or 50 minutes. (for 60 minutes, 70 minutes, 80 minutes, or 90 minutes), or increasing or continuous This includes administration by continuous intravenous administration.

[0131] The precise dosage and duration of treatment should be correlated with the disease being treated. Furthermore, empirically using known test protocols, or using methods known in the art This can be determined by testing the composition in the Dell system and then extrapolating from there. A bed test may also be conducted. The dosage will also vary depending on the severity of the condition to be alleviated. To obtain. Pharmaceutical compositions are generally therapeutically useful while minimizing undesirable side effects. It is formulated and administered to produce the desired effect. The composition is administered once, or multiple times. It may be divided into several smaller doses and administered at time intervals. Regarding elephants, the specific dosage regimen will depend on the individual's needs. It can be adjusted over time.

[0132] A therapeutic or pharmaceutical composition according to certain embodiments of the present disclosure, containing the active The component is bioavailable through administration to the subject or patient of the composition. It is formulated to enable this. The composition administered to the target or patient may be one or more. It can take the form of several dosage units. Practical methods for preparing such dosage forms are known to those skilled in the art. It is, or will be clear; for example, Remington: The Science and Practice of Pha See rmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The compositions that may be administered are typically for the treatment of the disease or condition described herein. It contains therapeutically effective amounts of the listed active ingredients.

[0133] In a particular embodiment, a polyomavirus neutralizing antibody or its antigen-binding fragment The dosage, for example, the dosage in step (a), is approximately 1 to approximately 100 mg / kg, or Approximately 10-30 mg / kg, or approximately 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5mg / kg, 6mg / kg, 7mg / kg, 8mg / kg, 9mg / kg, 1 0mg / kg, 15mg / kg, 20mg / kg, 25mg / kg, 30mg / kg, 3 5mg / kg, 40mg / kg, 45mg / kg, 50mg / kg, 60mg / kg, 7 The dosage is 0 mg / kg, 80 mg / kg, 90 mg / kg, or 100 mg / kg. In some embodiments, the further dose in step (c) is the same as in (a). The dosage may be the same as or different from the prescribed dosage, for example, approximately 1 to 100 mg / kg, or approximately 10 to 30 mg / kg. g / kg, or approximately 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6mg / kg, 7mg / kg, 8mg / kg, 9mg / kg, 10mg / kg, 15mg / kg, 20mg / kg, 25mg / kg, 30mg / kg, 35mg / kg, 40mg / kg, 45mg / kg, 50mg / kg, 60mg / kg, 70mg / kg, The dosage is 80 mg / kg, 90 mg / kg, or 100 mg / kg. Several implementations In the state, the time between (a) and (c) is approximately or at least approximately 1 day, 2 days, 3 days. Intervals: 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days 13 days, 14 days, 15 days, 15 days, 17 days, 18 days, 19 days, 20 days , 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days 29 days, or 30 days, or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 Weekly, 7 weeks, or 8 weeks, or 1 month, 2 months, or 3 months, Approximately 1 day or less, 2 days or less, 3 days or less, 4 days or less, 5 days or less, 6 days or less, 7 days Less than 8 days, 9 days or less, 10 days or less, 11 days or less, 12 days or less, 13 days Less than 14 days, 15 days or less, 15 days or less, 17 days or less, 18 days or less, 1 9 days or less, 20 days or less, 21 days or less, 22 days or less, 23 days or less, 24 days or less , 25 days or less, 26 days or less, 27 days or less, 28 days or less, 29 days or less, or 30 days or less, or 1 week or less, 2 weeks or less, 3 weeks or less, 4 weeks or less, 5 weeks or less 6 weeks or less, 7 weeks or less, or 8 weeks or less, or 1 month or less, or 2 months or less or less than 3 months.

[0134] In some embodiments, a polyomavirus neutralizing antibody or its antigen-binding fragment The dosage is approximately 10-30 mg / kg, or approximately 10-25 mg / kg, or 10-20 mg / kg, 10~15mg / kg, 15~30mg / kg, 15~25mg / kg, 15~ 20 mg / kg, 20-30 mg / kg, or 25-30 mg / kg, or approximately 10 mg / kg, 11mg / kg, 12mg / kg, 13mg / kg, 14mg / kg, 15 mg / kg, 16mg / kg, 17mg / kg, 18mg / kg, 19mg / kg, 20 mg / kg, 21mg / kg, 22mg / kg, 23mg / kg, 24mg / kg, 25 mg / kg, 26mg / kg, 27mg / kg, 28mg / kg, 29mg / kg, if The dosage is 30 mg / kg. In some cases, a dosage of 10-30 mg / kg is used in Invitational. When measured in a viral assay, BK or JC polio Optimal neutralizing activity (IC) of the target serum against the cytovirus 50 ) provides (for example) See Figure 2. In that case, the neutralizing activity increases up to a dose of 30 mg / kg, and the maximum neutralizing activity is, (This has been proven to be achieved at doses >10 mg / kg to <30 mg / kg.) Exemplary in vitro or ex vivo methods for measuring BK or JC virus replication For example, Abend et al. (J Virol. 81(1):272-279, 2007) and Low et al. ( As described in Virology. 323:182-8, 2004, the primary renal proximal tubular epithelium (RPTE) ) The use of primary renal proximal tubular epithelial (RPTE) cells grown and maintained in cell culture medium is one example. In specific embodiments, the drug is administered intravenously or subcutaneously.

[0135] In a particular embodiment, the administration regimens and methods described herein are one Alternatively, characterized by multiple pharmacokinetic profiles (including Table E2, see Examples). (See reference). For example, after administration of a polyomavirus neutralizing antibody or its antigen-binding fragment, At least 1%, 2%, 3%, 4% of the circulating serum concentration of the body or its antigen-binding fragments, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 1 6%, 17%, 18%, 19%, or 20%, or more, are infected organs, i.e. It penetrates the interstitial space of organs infected with BK virus or JC virus. Examples of infected organs include Examples include the bladder, kidneys, and brain. In some embodiments, antibodies or The average clearance of antigen-binding fragments is approximately 0.0760–0.0996 mL / day / kg. Yes. In some embodiments, the average distribution volume of an antibody or its antigen-binding fragment is The concentration is approximately 49.8–81.9 mL / kg.

[0136] Combination therapies are also included, for example, polyomavirus neutralizing antibodies or their antigen-binding determination. One of these is administered in combination with additional active ingredients such as immunosuppressants. Therefore, in some cases, A specified pharmaceutical or therapeutic composition further comprises additional active ingredients, such as immunosuppressants. Examples of immunosuppressants include monophosphate dehydrogenase inhibitors, purine synthesis inhibitors, and calcium phosphate inhibitors. Phosphate inhibitors, mTOR inhibitors, mycophenolate mofetil (MMF), mycophenol Sodium ruate, azathioprine, tacrolimus, sirolimus, and cyclosporine These are some examples.

[0137] The combination therapies described herein involve polyomavirus neutralizing antibodies or their antigen-binding properties. A single pharmaceutical formulation containing fragments and additional active ingredients. Administration of (ation), in addition, polyomavirus neutralizing antibody or its antigen-binding fragment and The administration of compositions containing additional active ingredients, each of which is itself in a separate pharmaceutical preparation. It is possible. For example, polyomavirus neutralizing antibodies or their antigen-binding fragments and additional The active ingredients may be administered together to the subject in a single-dose composition, or each active ingredient may be administered separately. It can be administered as a drug formulation. For example, a polyomavirus neutralizing antibody or its anti- The primordial fragment and additional therapeutic agents are placed in saline or other physiologically acceptable solutions, etc. In a single parenteral drug composition, these can be administered together to the subject, or each active ingredient can be administered separately. It can be administered as a parenteral medication. When separate medications are used, the composition is At the same time qualitatively (i.e., simultaneously), or at differently staggered times (i.e., sequentially) They may be administered secondarily and in any order. Combination therapy includes all of these regimens. This is how it is understood.

[0138] In a particular embodiment, the administration regimen of the treatment described herein The method involves BK or JC viremia and / or viral infection in the subject. For example, urine disease compared to a control (e.g., without composition or with polyomavirus neutralizing antibody) (or before treatment with the antigen-binding fragment), approximately or at least about 10%, 20 %, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 1000%, 2000%, 3000%, 4000%, Or reduce by 5000% or more. In some embodiments, as specified herein The administration regimen or method of the treatment described herein involves one or more BKs in the subject. These are symptoms related to the JC virus, such as blurred vision or other visual changes, brown or red urine, and urination. Painful urination, decreased kidney function, difficulty urinating, cough, common cold, shortness of breath, fever, muscle pain, muscle weakness, and / or reduces or improves symptoms such as seizures. [Examples]

[0139] [Example 1] Preclinical characterization of MAU868 (broad-spectrum neutralizing antibody against BK virus) To characterize the in vitro binding activity and virus neutralizing activity of MAU868, Procedure (i) was performed. Binding affinity was determined using a solution equilibrium titration assay. Primary renal proximal urine. Neutralization of BKV infection in tubular epithelial cells (RPTE) using immunofluorescence-based high-container TAg-expressing cells were evaluated by quantifying them using a CT imaging assay. The emergence of BKV-resistant variants (RAVs) with reduced susceptibility to AU868 indicates Two findings regarding BKV genotypes I and IV in RPTE and HEK-293 cells This was investigated in a long-term selection study. The MAU868 single-strand variable fragment (scFv) bound to the VP1 pentamer was examined. Crystallographic studies were conducted using ).

[0140] MAU868 has pM binding affinity and less than nM for four major BKV genotypes. It has neutralizing activity and EC 50 and EC 90 The values ​​range from 0.009 μg / ml each. Up to 0.093 μg / ml (0.062~0.645 nM) and 0.102~4.16 The range is 0 μg / ml (0.708 to 28.865 nM) (see, for example, Table E1 below). (see).

[0141] [Table 4]

[0142] No cytotoxicity was observed up to the highest concentration tested (500 μg / ml). MAU 868 is also a clinical isolate or a highly prevalent VP1 gene polymorphism, and JC Wi BKV virus constructed to contain the VP1 sequence derived from Rus (a closely related polyomavirus) It strongly neutralized riant. In the presence of MAU868, BKV continued for up to 182 days. After passage, RAV was not identified. Complex with VP1 pentamer at 2.66 Å resolution. The crystal structure of MAU868 in is strictly conserved across BKV isolates, and MA VP1 is involved in U868's broad-spectrum activity and its high barrier against resistance. The three-dimensional epitope containing the three contact residues (Y169, R170, K172) in Identified: Double or triple alanine at residues Y169, R170, or K172. BKV variants with substitutions were unable to grow.

[0143] A powerful broad-spectrum anti-whammy combined with a high barrier against resistance in vitro. Illus activity is the primary therapeutic agent for the treatment or prevention of BKV-related diseases (MAU). This demonstrates the potential of the 868.

[0144] [Example 2] Human research on MAU868 A randomized, blinded, placebo-controlled, single-dose escalation study was conducted at a single center in the United States. The study was blinded to participants, researchers, and sponsors. It was a randomized, placebo-controlled, blinded study. In a single-dose dose escalation design, MAU868 was administered iv(1 mg / mg) to healthy adults. (kg, 3 mg / kg, 10 mg / kg, 30 mg / kg, and 100 mg / kg) Each patient was administered sc (3 mg / kg). Each IV cohort consisted of 5 subjects (4 MA patients). U868: 1 person (placebo); the sc cohort consisted of 8 subjects (6 MAU) The ratio was 868:2 (placebo). The subjects underwent daily monitoring for 24 hours, followed by 106 days. Overall monitoring and pharmacokinetic (PK) evaluation were observed.

[0145] The serum concentration of MAU868 was measured using a verified sandwich ELISA with colorimetric detection. Determined by an assay based on the following criteria. MAU868 plasma concentration-time data were obtained from WinNo Typical exposure measurements were performed by analyzing using the standard non-compartment method with NLIN. Values ​​(AUC, Cmax, Tmax, elimination half-life) were obtained. Dose-proportionality was determined by log-converted dose. This was investigated by regression of log-transformed Cmax and AUC.

[0146] The ex vivo neutralizing activity of serum was measured in primary renal proximal urine before dose administration and 4 weeks after dose administration. Primary renal proximal tubular epithelium (RPT) grown and maintained in tubular epithelial (RPTE) cell culture medium E) Measurements were taken using cells.

[0147] 33 participants completed this study. Adverse events were mild and rare; more than one participant. The adverse events that occurred were nasal congestion (3 people, 9.1%) and oropharyngeal pain (3 people, 9.1%). This included bleeding at the injection site (post-injection hemorrhage; 2 people, 6.1%). There were no responses. No participants discontinued the study due to adverse events or the development of anti-drug antibodies.

[0148] Exemplary PK characteristics are shown in Figure 1 and Table E2 below.

[0149] [Table 5]

[0150] MAU868 PK had a half-life of 23-30 days. AUC and Cmax were: The dosage is proportional, ranging from 9880 μg / hour / mL to 1060000 μg / hour / mL. The levels were in the range of 24.7–2740 μg / mL, and there was no evidence of FcRn saturation. Plasma MAU8 on day 29, adjusted for extravascular distribution to estimate actual exposure. 68 concentration is the highest in vitro EC 50 (0.093 μg / mL) is approximately 7 to 751 times Yes. After IV administration, the mean clearance of MAU868 was 0.0760 mL / day / The range was between kg and 0.0996 mL / day / kg; the mean distribution volume was 49.8 m³. The range was between L / kg and 81.9 mL / kg. For both administration routes, AUC The coefficients of variation for and Cmax were ≤28% and ≤34%, respectively. c. Post-injection bioavailability was 57.6% during the maximum serum ex vivo period. The activity was achieved for doses >10 mg / kg (see Figure 2), and for doses between 10 and 30 mg / kg It appeared to be optimal within the administration range.

[0151] The MAU868 was safe and well-tolerated. PK data, for example, minimum C tro ughBy monitoring and defining the optimal blood flow rate for MAU898, the administration regimen can be determined. Evidence of maintaining concentrations in the clean / tissue, ex vivo neutralizing activity, is used in the treatment of BKV disease. Alternatively, it suggests the optimal scope of treatment for prevention.

Claims

1. For the treatment of BK or JC polyomavirus infections in human subjects that require it. This is a dosage regimen, (a) an antibody that specifically binds to the VP1 protein of the polyomavirus or A step of administering a dosage of antigen-binding fragments to the subject parenterally; (b) Serum or tissue concentration of the antibody or its antigen-binding fragment in the subject. Steps to measure; and (c) Trough concentration in serum or tissue (C ｔｒｏｕｇｈ ) is approximately 3 to 860 μm Before lowering the concentration to less than g / mL, administer further doses of the antibody or its antigen-binding fragment. Steps to cooperate Includes, The administration regimen concentrates the antibody or its antigen-binding fragment in the serum or tissue. The degree, throughout the treatment, C ｔｒｏｕｇｈ A dosage regimen to maintain a higher level.

2. (i)(c) C ｔｒｏｕｇｈ This is about plasma, and the administration registration Men, plasma C ｔｒｏｕｇｈ Before it drops to less than approximately 150-860 μg / mL, the above The step includes administering a certain dosage; (ii)(c) C ｔｒｏｕｇｈ This is about kidney tissue, and the administration The regimen is the C in renal tissue. ｔｒｏｕｇｈ The concentration is approximately 23.5 to less than 120 μg / mL. The step includes administering the additional dose before the descent; or (iii)(c) C ｔｒｏｕｇｈ This pertains to bladder tissue, and the above The administration regimen is the C in bladder tissue. ｔｒｏｕｇｈ The concentration drops to approximately 3-10 μg / mL or less. The step includes administering the additional dose before defecation, The administration regimen according to claim 1.

3. The antibody or its antigen-binding fragment Complementary determining regions (CDRs) V of SEQ ID NOs: 6-8, respectively Ｈ CDR1, V Ｈ CDR2, and Call V Ｈ Heavy chain variable (V) containing the CDR3 sequence Ｈ ) domain; and V corresponding to sequence numbers 9-11 Ｌ CDR1, V Ｌ CDR2 and V Ｌ CDR3 sequence Including light chain variable (V Ｌ )region, as well as those variants that specifically bind to the VP1 protein The administration regimen according to claim 1 or 2, comprising:

4. The aforementioned V Ｈ The region is at least 80%, 85%, 90%, 95%, and 97% of the sequence number 12. , containing 98%, 99%, or 100% identical sequences, optionally, V Ｈ The array is, Select one of the following: V5Q, G9P, T10G, N30S, N30K, and N30Q Multiple options can be selected, with 1, 2, 3, 4, 5, or 6 variations in the frame. It is located in the area of ​​the quad; and The aforementioned V Ｌ The region is at least 80%, 85%, 90%, 95%, and 97% of sequence number 13. , containing 98%, 99%, or 100% identical sequences, optionally, V Ｌ The array is 1 The framework has two, three, four, five, or six changes within its domain. The administration regimen according to claim 3.

5. The aforementioned V Ｈ The region contains, consists of, or is essentially derived from sequence number 12. and the aforementioned V Ｌ The region contains, consists of, or is essentially derived from sequence number 13. The administration regimen according to claim 2 or 3.

6. The aforementioned administration regimen controls the BK virus genotype or JC virus in the subject. A dosage regimen according to any one of claims 1 to 5, comprising the step of determining

7. The polyomavirus infection is one of genotypes I, II, III, and IV or It contains BK viruses selected from multiple sources, or the polyomavirus infection is JC virus A dosage regimen according to any one of claims 1 to 6, comprising Rus.

8. The administration regimen according to any one of claims 1 to 7, wherein the subject is immunocompromised.

9. The aforementioned subjects perform transplantation procedures, optionally organ transplantation or cell-based transplantation procedures, precisely Claims 1 to 8: The person who is about to receive, is in the process of receiving, or has received The administration regimen described in any one of the items.

10. The transplantation procedure is selected from kidney transplantation and hematopoietic stem cell transplantation (HCT), as described in claim 9. The listed administration regimen.

11. The aforementioned subjects are BK virus-associated nephropathy, BK virus-associated hemorrhagic cystitis, and JC virus Having or being at risk of having a condition selected from Ruth-associated progressive multifocal leukoencephalopathy, The administration regimen according to any one of claims 1 to 10.

12. (a) The dosage of the drug in the above is approximately 1 to approximately 100 mg / kg or approximately 10 to 30 mg / kg kg, or approximately 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg g, 6mg / kg, 7mg / kg, 8mg / kg, 9mg / kg, 10mg / kg, 15 mg / kg, 20mg / kg, 25mg / kg, 30mg / kg, 35mg / kg, 40 mg / kg, 45mg / kg, 50mg / kg, 60mg / kg, 70mg / kg, 80 The dosage is mg / kg, 90 mg / kg, or 100 mg / kg, and the aforementioned medication is administered at the discretion of the user. The dosage regimen according to any one of claims 1 to 11, wherein the amount is administered intravenously or subcutaneously. hmm.

13. (c) The further dose in (c) is the same as or different from the dose in (a), Selectively, approximately 1 to 100 mg / kg, approximately 10 to 30 mg / kg, or approximately 1 mg / kg, 2mg / kg, 3mg / kg, 4mg / kg, 5mg / kg, 6mg / kg, 7 mg / kg, 8mg / kg, 9mg / kg, 10mg / kg, 15mg / kg, 20mg / kg, 25mg / kg, 30mg / kg, 35mg / kg, 40mg / kg, 45mg / kg, 50mg / kg, 60mg / kg, 70mg / kg, 80mg / kg, 90mg The dosage is either / kg or 100 mg / kg, and optionally, the dosage can be administered intravenously or subcutaneously. An administration regimen according to any one of claims 1 to 12, administered to [the patient].

14. The polyomavirus infection includes BK virus genotype I, and here, Said C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, and optionally the kidney tissue concentration, is the antibody concentration in the above-mentioned area. or the EC of its antigen-binding fragment ５０ From approximately 2618 times to approximately 3775 times, approximately 13,06 It is within the range of 1x; or Said C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, and optionally the bladder tissue concentration, is the antibody or the EC of its antigen-binding fragment ５０ From approximately 500 times to approximately 692 times, and up to approximately 1000 times. This is within the range, The EC ５０ The concentration is approximately 0.009 ± 0.010 μg / mL. The administration regimen according to any one of claims 1 to 13.

15. The polyomavirus infection includes BK virus genotype II, and here, C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, or optionally the renal tissue concentration, is the antibody or The EC of that antigen-binding fragment ５０ From approximately 589 times to approximately 849 times, and up to approximately 2942 times. (For example, approximately 600 times, 700 times, 800 times, 900 times, 1000 times, 1500 times, 2 (000 times, 2500 times, 3000 times) and; or C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, and optionally the bladder tissue concentration, the antibody or The EC of its antigen-binding fragment ５０ From approximately 100 times to approximately 156 times, and up to approximately 200 times. It is an enclosure, The EC ５０ The concentration is approximately 0.040 ± 0.025 μg / mL. The administration regimen according to any one of claims 1 to 13.

16. The polyomavirus infection includes BK virus genotype III, and here, Said C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, and optionally the kidney tissue concentration, is the antibody concentration in the above-mentioned area. or the EC of its antigen-binding fragment ５０ From approximately 253 times to approximately 365 times, and up to approximately 1265 times. It is within the range; or Said C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, and optionally the bladder tissue concentration, is the antibody or the EC of its antigen-binding fragment ５０ From approximately 50 times to approximately 67 times, up to approximately 100 times. (For example, approximately 50 times, 55 times, 60 times, 65 times, 70 times, 75 times, 80 times, 85 times, 9 (0x, 95x, or 100x) The EC ５０ The concentration is approximately 0.093 ± 0.057 μg / mL. The administration regimen according to any one of claims 1 to 13.

17. The polyomavirus infection includes BK virus genotype IV, and here, Said C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, and optionally the kidney tissue concentration, is the antibody concentration in the above-mentioned area. or the EC of its antigen-binding fragment ５０ From approximately 1122 times to approximately 1618 times, and approximately 5604 times It is within the range of; or Said C ｔｒｏｕｇｈ The tissue concentration in the above-mentioned area, and optionally the bladder tissue concentration, is the antibody or the EC of its antigen-binding fragment ５０ From approximately 100 times to approximately 297 times, up to approximately 500 times It is within the range, The EC ５０ The concentration is approximately 0.021 ± 0.020 μg / mL. The administration regimen according to any one of claims 1 to 13.

18. The polyomavirus infection includes the JC virus, and the C ｔｒｏｕｇｈ The above The tissue concentration is the EC of the antibody or its antigen-binding fragment. ５０ At least approximately 29 times Up to approximately 110 times, up to approximately 158 times, and up to approximately 547 times, and the aforementioned EC ５０ approximately 0.215 The dosage regimen according to any one of claims 1 to 13, wherein the concentration is ±0.130 μg / mL.

19. The time between (a) and (b) is approximately or at least approximately 1 day, 2 days, 3 days, 4 days, and 5 days. 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 14th, 16th , 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th , the 27th, 28th, 29th, or 30th, or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, or 1 month, 2 months, or 3 months, or about 1 day Below, 2 days or less, 3 days or less, 4 days or less, 5 days or less, 6 days or less, 7 days or less, 8 days or less, 9 days or less Below, 10 days or less, 11 days or less, 12 days or less, 13 days or less, 14 days or less, 14 days or less, 16 days or less, 17 days or less, 18 days or less, 19 days or less, 20 days or less, 21 days or less, 22 days or less, 23 days or less, 24 days or less, 25 days or less, 26 days or less, 27 days or less, 28 days or less, 29 days or less Below, or 30 days or less, or 1 week or less, 2 weeks or less, 3 weeks or less, 4 weeks or less, 5 weeks or less , less than 6 weeks, less than 7 weeks, or less than 8 weeks, or less than 1 month, less than 2 months, or A dosage regimen according to any one of claims 1 to 18, wherein the duration is three months or less.

20. (b) In the subjects mentioned above, serum or tissue levels were increased for approximately 3, 4, 5, 6, and 7 days. Sunday, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 14th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, Once every 28th, 29th, or 30th, or approximately every 1, 2, 3, 4, 5, or 6 weeks. , 1 to 3 times every 7 or 8 weeks, or 1 to 6 times every 1, 2, or 3 months. A dosage regimen according to any one of claims 1 to 19, comprising the step of measuring.

21. The time between (a) and (c) is approximately or at least approximately 1 day, 2 days, 3 days, 4 days, and 5 days. 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 14th, 16th , 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th , the 27th, 28th, 29th, or 30th, or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, or 1 month, 2 months, or 3 months, or about 1 day Below, 2 days or less, 3 days or less, 4 days or less, 5 days or less, 6 days or less, 7 days or less, 8 days or less, 9 days or less Below, 10 days or less, 11 days or less, 12 days or less, 13 days or less, 14 days or less, 14 days or less, 16 days or less, 17 days or less, 18 days or less, 19 days or less, 20 days or less, 21 days or less, 22 days or less, 23 days or less, 24 days or less, 25 days or less, 26 days or less, 27 days or less, 28 days or less, 29 days or less Below, or 30 days or less, or 1 week or less, 2 weeks or less, 3 weeks or less, 4 weeks or less, 5 weeks or less , less than 6 weeks, less than 7 weeks, or less than 8 weeks, or less than 1 month, less than 2 months, or A dosage regimen according to any one of claims 1 to 20, wherein the duration is three months or less.

22. At least 1%, 2%, or 3% of the circulating concentration of the antibody or its antigen-binding fragment in serum. 、４％、５％、６％、７％、８％、９％、１０％、１１％、１２％、１３％、１４％、１ 5%, 16%, 17%, 18%, 19%, or 20%, or more, immersed in the infected organ. The administration regimen according to any one of claims 1 to 21, wherein the agent penetrates the substance.

23. Claim 22, wherein the infected organ is the bladder, kidney, or central nervous system (CNS). The administration regimen.

24. The average clearance of the antibody or its antigen-binding fragment is approximately 0.0760 to 0.

099. The administration regimen according to any one of claims 1 to 23, wherein the dose is 6 mL / day / kg.

25. The average distribution volume of the antibody or its antigen-binding fragment is approximately 49.8 to 81.9 mL / kg. The administration regimen according to any one of claims 1 to 24.

26. To treat BK or JC polyomavirus infections in human subjects who require it. It is a method for that purpose, An antibody or antigen that specifically binds to the VP1 protein of the polyomavirus. The step includes administering a dosage of the combined fragment to the subject parenterally, wherein the dosage is approximately 1 0-30 mg / kg, or approximately 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17mg / kg, 18 mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22mg / kg, 23 mg / kg, 24mg / kg, 25mg / kg, 26mg / kg, 27mg / kg, 28 A method in which the dose is mg / kg, 29 mg / kg, or 30 mg / kg.

27. The antibody or its antigen-binding fragment Complementarity Determination Region (CDR) V for sequence numbers 6-8, respectively. Ｈ CDR1, V Ｈ CDR2, Call V Ｈ Heavy chain variable (V) containing the CDR3 sequence Ｈ ); and V corresponding to sequence numbers 9-11 Ｌ CDR1, V Ｌ CDR2 and V Ｌ CDR3 sequence Including light chain variable (V Ｌ )region, as well as those variants that specifically bind to the VP1 protein The method according to claim 26, including the method described in claim 26.

28. The aforementioned V Ｈ The region is at least 80%, 85%, 90%, 95%, and 97% of the sequence number 12. It contains 98%, 99%, or 100% identical sequences, and optionally, the V Ｈ The array is arbitrary. Select one or more of the following: V5Q, G9P, T10G, N30S, N30K, and N30Q. A framework of one, two, three, four, five, or six variations, selected from a number. Having in the domain; and The aforementioned V Ｌ The region is at least 80%, 85%, 90%, 95%, and 97% of the sequence number 13. It contains 98%, 99%, or 100% identical sequences, and optionally, the V Ｌ There is one array. , having two, three, four, five, or six changes in the framework domain, The method according to claim 27.

29. The aforementioned V Ｈ The region contains, consists of, or is essentially derived from sequence number 12. and the aforementioned V Ｌ The region contains, consists of, or is essentially derived from sequence number 13. The method according to claim 27 or 28.

30. The polyomavirus infection is one of genotypes I, II, III, and IV or The method according to any one of claims 26 to 29, comprising a BK virus selected from a plurality. 。

31. The polyomavirus infection includes the JC virus, any one of claims 26 to 30. The method is described below.

32. The method according to any one of claims 26 to 31, wherein the subject is immunocompromised.

33. The aforementioned subjects perform transplantation procedures, optionally organ transplantation or cell-based transplantation procedures, precisely Claims 26-3: In the process of receiving, in the process of receiving, or have received The method described in any one of item 2.

34. Claim 33, wherein the transplantation procedure is selected from kidney transplantation and hematopoietic stem cell transplantation (HCT). Method of description.

35. The aforementioned subjects are BK virus-associated nephropathy, BK virus-associated hemorrhagic cystitis, and JC virus Having or being at risk of having a condition selected from Ruth-associated progressive multifocal leukoencephalopathy, The method according to any one of claims 26 to 34.

36. At least 1%, 2%, or 3% of the circulating concentration of the antibody or its antigen-binding fragment in serum. 、４％、５％、６％、７％、８％、９％、１０％、１１％、１２％、１３％、１４％、１ 5%, 16%, 17%, 18%, 19%, or 20%, or more, immersed in the infected organ. The method according to any one of claims 26 to 35, which allows for penetration.

37. The method according to claim 36, wherein the infected organ is the bladder or the kidney.

38. The average clearance of the antibody or its antigen-binding fragment is approximately 0.0760 to 0.

099. The method according to any one of claims 26 to 37, wherein the amount is 6 mL / day / kg.

39. The average distribution volume of the antibody or its antigen-binding fragment is approximately 49.8 to 81.9 mL / kg. The method according to any one of claims 26 to 38.

40. Dosages of 10-30 mg / kg are used in in vitro or ex vivo viral assays. When measured in the aforementioned serum, the BK or JC polyomavirus The optimal neutralizing activity (IC) ５０ The administration method according to any one of claims 1 to 39, which gives ) Method or technique.

41. In the aforementioned subjects, BK or JC viremia and / or viruria are selected at will. In the selection, approximately or less than before treatment with the control or antibody or its antigen-binding fragment. Approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 1 ００％、２００％、３００％、４００％、５００％、１０００％、２０００％、３０００ To reduce by %, 4000%, or 5000%, or more, according to claims 1 to 40. Any of the administration regimens or methods described in item one.

42. In the aforementioned subjects, one or more BK or JC virus-related symptoms, optionally selected, fog Vision or other visual changes, brown or red urine, painful urination, decreased kidney function, difficulty urinating, cough, One of the following symptoms may be selected from: cold, shortness of breath, fever, muscle pain, muscle weakness, and / or seizures. It reduces kidney inflammation such as ureteral stricture or interstitial nephritis, and is used selectively for multiple symptoms. The administration regimen or method according to any one of claims 1 to 41 is improved.

43. Histidine, sugars, optionally sucrose, and polyols, optionally polysorbates. The step of administering the antibody or its antigen-binding fragment in a pharmaceutical composition containing the antibody Including the administration regimen or method according to any one of claims 1 to 42.

44. Formulated for parenteral administration at a dosage of approximately 10-30 mg / kg, and each is distributed Complementarity Determination Region (CDR) V in columns 6-8 Ｈ CDR1, V Ｈ CDR2 and V Ｈ CD Heavy chain variable (V) containing R3 sequence Ｈ ), and V of sequence numbers 9 to 11, respectively. Ｌ CDR1, V Ｌ CDR2 and V Ｌ Light chain variable (V) containing CDR3 sequence Ｌ ) an antibody or including the region Its antigen-binding fragment; and Histidine, sugars, optionally sucrose, and polyols, optionally polysorbates. Medicinal carriers including A pharmaceutical composition containing the following:

45. The aforementioned V Ｈ The region is at least 80%, 85%, 90%, 95%, and 97% of the sequence number 12. , containing 98%, 99%, or 100% identical sequences, optionally, V Ｈ The array is, Select one of the following: V5Q, G9P, T10G, N30S, N30K, and N30Q Multiple options can be selected, with 1, 2, 3, 4, 5, or 6 variations in the frame. It is located in the area of ​​the quad; and The aforementioned V Ｌ The region is at least 80%, 85%, 90%, 95%, and 97% of sequence number 13. , containing 98%, 99%, or 100% identical sequences, optionally, V Ｌ The array is 1 The framework has two, three, four, five, or six changes within its domain. The pharmaceutical composition according to claim 44.

46. The antibody or its antigen-binding fragment is approximately 10 mg / kg, 11 mg / kg, or 12 mg / kg, 13mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17mg / kg, 18mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22mg / kg, 23mg / kg, 24mg / kg, 25mg / kg, 26mg / kg, 27mg Parenteral administration at doses of 28 mg / kg, 29 mg / kg, or 30 mg / kg. A pharmaceutical composition according to claim 44 or 45, formulated for administration.

47. A pharmaceutical product formulated for intravenous administration, as described in any one of claims 44 to 46. composition.

48. A pharmaceutical compound formulated for subcutaneous administration, according to any one of claims 44 to 46 A finished product.

49. To treat BK or JC polyomavirus infections in human subjects who require it. A pharmaceutical composition according to any one of claims 44 to 48 for use in the foregoing.

50. The polyomavirus infection is one of genotypes I, II, III, and IV or A pharmaceutical composition for use according to claim 49, comprising a BK virus selected from a plurality of others.

51. The use according to claim 49 or 50, wherein the polyomavirus infection includes the JC virus. A pharmaceutical composition for use.

52. The subject is immunocompromised, for use according to any one of claims 49 to 51 Pharmaceutical composition.

53. The aforementioned subjects perform transplantation procedures, optionally organ transplantation or cell-based transplantation procedures, precisely Claims 49-5: In the process of receiving, receiving, or have received A pharmaceutical composition for use as described in any one of paragraphs 2.

54. Claim 53, wherein the transplantation procedure is selected from kidney transplantation and hematopoietic stem cell transplantation (HCT). A pharmaceutical composition for the use described.

55. The aforementioned subjects are BK virus-associated nephropathy, BK virus-associated hemorrhagic cystitis, and JC virus Having or being at risk of having a condition selected from Ruth-associated progressive multifocal leukoencephalopathy, A pharmaceutical composition for use according to any one of claims 49 to 54.

Images