Compositions and methods for cell depletion

Abstract

The present invention provides compositions and methods for promoting the engraftment of exogenous hematopoietic stem cell grafts and for maintaining the pluripotency and hematopoietic function of these cells after transplantation. [Solution] The present invention provides compositions and methods useful for the depletion of cells such as CD45+, CD135+, CD34+, CD90+, and / or CD110+ cells, and, in particular, for the treatment of various hematopoietic disorders, metabolic disorders, cancer, and autoimmune diseases. For example, antibodies, their antigen-binding fragments, ligands, and conjugates that can be applied to achieve the treatment of these conditions by depleting populations of D45+, CD135+, CD34+, CD90+, or CD110+ cells in a patient such as a human are described herein.

Description

[Technical Field]

[0001] This invention is particularly relevant to a variety of conditions such as blood disorders, metabolic disorders, cancer, and autoimmune diseases. In patients suffering from the disease, antigens expressed by hematopoietic cells such as hematopoietic stem cells bind to these antigens. Treatment involves the administration of antibodies, their antigen-binding fragments, or ligands. do. [Background technology]

[0002] Despite advances in medical technology, hematopoietic disorders, particularly those involving specific blood cells and metabolic disorders, remain prevalent. There remains a demand for treating diseases, cancer, and autoimmune conditions. Stem cells have great therapeutic potential, but the constraints that have hindered their clinical use include hematopoiesis. The difficulty lay in ensuring the successful engraftment of stem cell grafts into the host. Currently, exogenous hematopoietic stem cell grafts... To promote engraftment and ensure that the pluripotency and hematopoietic function of these cells are maintained after transplantation. Compositions and methods for this purpose are needed. [Overview of the Initiative]

[0003] This invention is particularly relevant to various hematopoietic disorders, metabolic disorders, cancer, and autoimmune diseases. The present invention provides compositions and methods for the direct treatment of hematopoietic stem cell transplantation therapy. Before receiving the procedure, patients such as human patients are conditioned to promote the engraftment of hematopoietic stem cells. It is characterized by a method of conditioning. Patients have abnormal hemoglobinosis or other hematopoietic conditions. These patients may have one or more blood disorders, and therefore hematopoietic stem cell transplantation is necessary. The key point is that, as described herein, hematopoietic stem cells are derived from a number of cell types of the hematopoietic lineage. It can differentiate and increase or regrow the cell type that is deficient in the patient. The present invention relates to hematopoietic cells such as cell surface hematopoietic cell antigens. Antibodies, antibody fragments, ligands, and drugs that can bind to proteins expressed by [unspecified]. Methods of treating patients with substance-antibody conjugates and drug-ligand conjugates are being developed. (i) abnormal blood cells, cancer cells, or autoimmune cells, such as CD45, CD135, CD34 By selectively depleting the cell population expressing CD90 and / or CD110, Among those described herein, in particular blood disorders, metabolic disorders, cancer, or autoimmune diseases. (ii) directly treating diseases such as, and / or (ii) endogenous hematopoietic stem cell populations within patients. The objective is to deplete the CD45, CD135, CD34, CD90, and / or CD110, and others. Among these cell types, cancer cells such as leukemia cells and T cell receptors that cross-react with autoantigens are particularly noteworthy. Because it can be expressed by autoimmune lymphocytes such as T cells that express the body, the activity of the former is It enables direct treatment of a wide range of disorders related to hematopoietic cells. Selective degeneration of hematopoietic stem cells. The latter activity, which is thirst, then creates a vacancy, which is then replaced by an exogenous substance (e.g., self, homogeneous system). It can be later supplemented by transplantation of hematopoietic stem cell grafts (or of the same lineage). Therefore, the present invention particularly addresses sickle cell anemia, thalassemia, Fanconi anemia, and Wiscott anemia. - Aldrich syndrome, adenosine deaminase deficiency - severe combined immunodeficiency, metachromatic leukodystrophy Sterophy, Diamond-Blackfan anemia, and Schwarman-Diamond syndrome , human immunodeficiency virus infection, and various hematopoietic disorders such as acquired immunodeficiency syndrome This invention provides a treatment for the condition, as well as methods for treating cancer and autoimmune diseases.

[0004] In the first aspect, the present invention provides a material that can bind to CD135 and conjugate to a cytotoxin. By administering an effective amount of the antibody or its antigen-binding fragment to a human patient, This provides a method for depleting a population of CD135+ cells.

[0005] In another embodiment, the present invention binds to CD135 before the patient receives a graft containing hematopoietic stem cells. It is possible to use antibodies conjugated to cytotoxins or their antigen-binding fragments By administering an effective dose, CD135+ cells in human patients requiring hematopoietic stem cell transplantation can be effectively controlled. This provides a method for depleting a population of cells.

[0006] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient A person who has an antibody or antigen that can bind to CD135 and is conjugated to a cytotoxin. The binding fragment was pre-administered in an amount sufficient to deplete the CD135+ cell population in the patient. It is characterized by the given method.

[0007] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. The law states that in human patients, the cytotoxin that can bind to CD135 is conjugated. The antibody or its antigen-binding fragment depletes the population of CD135+ cells in the patient. The process involves administering a sufficient amount, followed by administering a graft containing hematopoietic stem cells to the patient. The method is characterized by including this.

[0008] In some embodiments of the four aspects described above, the antibody or its antigen-binding fragment is , including the following Complementarity Determination Regions (CDRs): CDR-H1 having the amino acid sequence SYYMH (SEQ ID NO: 1); CDR-H2 having the amino acid sequence IINPSGGSTSYAQKFQG (SEQ ID NO: 2); CDR-H having amino acid sequence GVGAHDAFDI (SEQ ID NO: 3) or VVAAAVADY (SEQ ID NO: 4) 3; Amino acid sequence RSSQSLLHSNGNNYLD (SEQ ID NO: 5) or RSSQSLLHSNGYNYLD (SEQ ID NO: 6) CDR-L1 having; CDR-L2 having the amino acid sequence LGSNRAS (SEQ ID NO: 7); and CDR-L3 having the amino acid sequence MQGTHPAIS (SEQ ID NO: 8) or MQSLQTPFT (SEQ ID NO: 9) .

[0009] In some embodiments of the four aspects described above, the antibody or its antigen-binding fragment is , including the following CDRs: CDR-H1 having amino acid sequence SYAIS (SEQ ID NO: 10); CDR-H2 having the amino acid sequence GIIPIFGTANYAQKFQG (SEQ ID NO: 11); CDR-H3 having the amino acid sequence FALFGFREQAFDI (SEQ ID NO: 12); CDR-L1 having the amino acid sequence RASQSISSYLN (SEQ ID NO: 13); CDR-L2 having the amino acid sequence AASSLQS (SEQ ID NO: 14); and CDR-L3 possessing the amino acid sequence QQSYSTPFT (SEQ ID NO: 15).

[0010] In another embodiment, the present invention provides a patient with a human Flt3 ligand capable of binding to CD135. Alternatively, by administering an effective amount of its fragment, CD135+ cells in human patients It is characterized by a method of depleting the population.

[0011] In another aspect, the present invention provides the patient with CD135 before receiving a transplant containing hematopoietic stem cells. Administer an effective amount of a human Flt3 ligand or a fragment thereof that can bind to it. This depletes the population of CD135+ cells in human patients requiring hematopoietic stem cell transplantation. It is characterized by a method of [doing something].

[0012] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient A human Flt3 ligand, or a fragment thereof, that can bind to CD135, is used in patients. This method is characterized by pre-administering a sufficient amount to deplete the population of CD135+ cells. ru.

[0013] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method for providing a patient with a human Flt3 ligand capable of binding to CD135, or the Administer the fragment in an amount sufficient to deplete the population of CD135+ cells in the patient; continue The method is characterized by including the step of administering a graft containing hematopoietic stem cells to a patient.

[0014] In some embodiments of any of the four embodiments described above, a human Flt3 ligand or The fragments are isolated from human antibodies (e.g., IgG1, IgG2, IgG3, or IgG4). Covalently bound to Fc domains such as dimeric Fc domains (isolated from sotype human antibodies) In some embodiments, the Fc domain is a monomer Fc containing a single-stranded polypeptide chain. It is a domain. In some embodiments, it is a human Flt3 ligand or a fragment thereof. The N-terminus is bound to the Fc domain. In some embodiments, a human Flt3 ligand is used. The C-terminus of the fragment is bound to the Fc domain. The Fc domain is one or more It may be conjugated to the Flt3 ligand or a fragment of P. For example, Conjugates that may be used in conjunction with the methods described herein include each polypeptide of the Fc domain. A dimer Fc-type molecule in which the cytoplasmic chain is conjugated to a human Flt3 ligand or a fragment thereof. The main component is included. The Fc domain then contains the cytotoxins described herein (e.g., Pseudomonas). Exotoxins such as A, deBouganin, diphtheria toxin, α-amanitin, and saporin Maytansine, Maytansinoid, Auristatin, Anthracycline, Calicema Icin, Irinotecan, SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolo Benzodiazepine dimers, indolinobenzodiazepines, and indolinobenzodiazepines It may be conjugated to cytotoxins such as pin dimers or their variants.

[0015] In some embodiments of any of the four embodiments described above, a human Flt3 ligand or The fragment is a cytotoxin as described herein (e.g., Pseudomonas exotoxin A, deBouga). Nin, diphtheria toxin, amatoxins such as α-amanitin, saporins, maytansine, me Itansinoids, auristatin, anthracyclines, calicheamicin, irinoteca N, SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolobenzodiazepine 2 Merges, indolinobenzodiazepines, and indolinobenzodiazepine dimers, or It is covalently bound to cytotoxins such as those of their variants. In some embodiments, human Fl The N-terminus of the t3 ligand or its fragment is bound to a cytotoxin. In the application form, the C-terminus of the human Flt3 ligand or its fragment binds to the cytotoxin. Yes. The cytotoxin may then be conjugated to the Fc domain.

[0016] In some embodiments of any of the four embodiments described above, a human Flt3 ligand or A fragment is a human Flt3 ligand or one site of that fragment (e.g., human F The lt3 ligand or its fragment (N or C terminus) is covalently bound to the cytotoxin. , another site of the human Flt3 ligand or its fragment (for example, the human Flt3 ligand or It is covalently bonded to the Fc domain at the opposite end of the fragment.

[0017] In some embodiments of any of the four embodiments described above, the Fc domain is human IgG1 eye It is a sotype Fc domain. In some embodiments, the Fc domain is a human IgG2 isotype. The Fc domain is a human IgG3 isotype. This is the Fc domain. In some embodiments, the Fc domain is the human IgG4 isotype Fc domain. This is the main point.

[0018] In another embodiment, the present invention relates to a CD34 that can bind to and conjugate to a cytotoxin. By administering an effective amount of the antibody or its antigen-binding fragment, in human patients This method is characterized by depleting the population of CD34+ cells.

[0019] In another embodiment, the present invention relates to the binding of CD34 to a patient before the patient receives a graft containing hematopoietic stem cells. It is possible to have antibodies or antigen-binding fragments conjugated to cytotoxins. By administering an effective dose, CD34+ cells in human patients requiring hematopoietic stem cell transplantation It is characterized by a method of depleting the population.

[0020] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody that can bind to CD34 and is conjugated to a cytotoxin or its antigen-binding agent Ragment is administered beforehand in an amount sufficient to deplete the CD34+ cell population in the patient. We provide a method for doing so.

[0021] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method that involves administering to the patient an anti-inflammatory drug that can bind to CD34 and is conjugated with a cytotoxin. The body or its antigen-binding fragments are used to deplete the population of CD34+ cells in the patient. The process involves administering a sufficient amount, followed by administering a graft containing hematopoietic stem cells to the patient. It is characterized by a method of inclusion.

[0022] In another aspect, the present invention relates to a cytotoxin that can bind to CD90 and be conjugated. By administering an effective amount of the antibody or its antigen-binding fragment, in human patients This method is characterized by depleting the population of CD90+ cells.

[0023] In another aspect, the present invention relates to the binding of CD90 to a graft containing hematopoietic stem cells before the patient receives the graft. It is possible to have antibodies or antigen-binding fragments conjugated to cytotoxins. By administering the effective dose, CD90+ cells in human patients requiring hematopoietic stem cell transplantation It is characterized by a method of depleting the population.

[0024] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody that can bind to CD90 and is conjugated to a cytotoxin or its antigen-binding agent Ragment is administered beforehand in an amount sufficient to deplete the CD90+ cell population in the patient. We provide a method for doing so.

[0025] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method that involves administering to the patient an anti-inflammatory drug that can bind to CD90 and is conjugated with a cytotoxin. The body or its antigen-binding fragments are used to deplete the population of CD90+ cells in the patient. The process involves administering a sufficient amount, followed by administering a graft containing hematopoietic stem cells to the patient. It is characterized by a method of inclusion.

[0026] In another aspect, the present invention relates to a method that can bind to CD110 and conjugate to cytotoxins. By administering an effective amount of the antibody or its antigen-binding fragment to a human patient, This method is characterized by its ability to deplete a population of CD110+ cells.

[0027] In another embodiment, the present invention binds to CD110 before the patient receives a graft containing hematopoietic stem cells. It is possible to use antibodies conjugated to cytotoxins or their antigen-binding fragments By administering an effective dose, CD110+ cells in human patients requiring hematopoietic stem cell transplantation can be effectively controlled. It is characterized by a method of depleting a population of cells.

[0028] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a patient, wherein the patient is CD Antibodies that can bind to 110 and are conjugated to cytotoxins or their antigen-binding films The drug is administered beforehand in an amount sufficient to deplete the population of CD110+ cells in the patient. We provide a way to do so.

[0029] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. This method involves providing the patient with a cytotoxin that can bind to CD110 and is conjugated with it. The antibody or its antigen-binding fragment depletes the population of CD110+ cells in the patient. The process involves administering a sufficient amount, followed by administering a graft containing hematopoietic stem cells to the patient. The method is characterized by including this.

[0030] In some embodiments of any of the aforementioned aspects, an antibody, its antigen-binding fragment, Alternatively, cytotoxins conjugated to ligands include Pseudomonas exotoxin A, deBouganin. Diphtheria toxin, amatoxins such as α-amanitin, saporins, maytansine, may Tansinoids, auristatin, anthracyclines, calicheamicin, irinotecan SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolobenzodiazepine (2 doses) Indolinobenzodiazepine, or indolinobenzodiazepine dimer, These are their variants.

[0031] In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for treating, for example, a human patient who requires a hematopoietic stem cell transplantation. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for treating, for example, a human patient who requires a hematopoietic stem cell transplantation. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof.

[0032] In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof. In another aspect, the present invention provides a method for depleting a population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplantation by administering an effective amount of an antibody or an antigen-binding fragment thereof that can bind to CD45 and is conjugated to a cytotoxin before the patient receives a graft comprising hematopoietic stem cells. The cytotoxin can be, for example, Pseudomonas exotoxin A, deBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, maytansinoids, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, duocarmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, or indolinobenzodiazepine dimer, or variants thereof.

[0033] In another aspect, the present invention provides a method for treating, for example, a human patient who requires a hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody that can bind to CD45 and is conjugated to a cytotoxin or its antigen-binding agent Ragment is administered beforehand in an amount sufficient to deplete the CD45+ cell population in the patient. The method is characterized by the following. Cytotoxins include, for example, Pseudomonas exotoxin A, deBouganin, Diphtheria toxin, amatoxins such as α-amanitin, saporins, meitansine, meita Insinoids, auristatin, anthracyclines, calicheamicin, irinotecan, SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolobenzodiazepine dimer indolinobenzodiazepine, or indolinobenzodiazepine dimer, or so These could be variants.

[0034] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method for administering to human patients a substance that can bind to CD45, and which contains Pseudomonas exotoxin A, d eBouganin, diphtheria toxin, amatoxins such as α-amanitin, saporins, meitans N, meitansinoids, auristatin, anthracyclines, calicheamicin, iri Notecan, SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolobenzodiazepine Indolinobenzodiazepine dimer, or indolinobenzodiazepine dimer antibodies conjugated to cytotoxins such as their variants or their antigen-binding agents The process of administering lagment in an amount sufficient to deplete the CD45+ cell population in the patient. The method is characterized by the step of subsequently administering a graft containing hematopoietic stem cells to the patient.

[0035] In some embodiments of the four embodiments described above, CD45 is CD45RO.

[0036] In another embodiment, the present invention relates to an antibody or antigen-binding filament that can bind to CD45RO. By administering an effective dose of the drug, the population of CD45RO+ cells in human patients is depleted. Provide a method to make it happen.

[0037] In another embodiment, the present invention binds to CD45RO before the patient receives a graft containing hematopoietic stem cells. By administering an effective amount of the antibody or its antigen-binding fragment, it is possible to achieve this. Methods to deplete the CD45RO+ cell population in human patients requiring hematopoietic stem cell transplantation. provide.

[0038] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody or antigen-binding fragment that can bind to CD45RO in a patient This method is characterized by pre-administering a sufficient amount of CD45RO+ cells to deplete the population.

[0039] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method for providing a patient with an antibody or antigen-binding flag that can bind to CD45RO. The process involves administering ment in an amount sufficient to deplete the population of CD45RO+ cells in the patient. The method is characterized by the step of subsequently administering a graft containing hematopoietic stem cells to the patient.

[0040] In some embodiments of any of the four aforementioned aspects of the present invention, an antibody or its antigen The conjugate fragment is conjugated to a cytotoxin.

[0041] In another aspect, the invention provides a method of depleting the population of CD45+ cells in a human patient by administering an effective amount of a ligand or fragment thereof that can bind to CD45.

[0042] In another aspect, the invention provides a method of depleting the population of CD45+ cells in a human patient who requires a hematopoietic stem cell transplant by administering an effective amount of a ligand or fragment thereof that can bind to CD45 before the patient receives a graft comprising hematopoietic stem cells.

[0043]

[0044] In another aspect, the invention provides a method of treating a human patient who requires a hematopoietic stem cell transplant, the method comprising administering to the human patient a graft comprising hematopoietic stem cells, wherein the patient has been previously administered a ligand or fragment thereof that can bind to CD45 in an amount sufficient to deplete the population of CD45+ cells in the patient.

[0045] In a further aspect, the invention provides a method of treating a human patient who requires a hematopoietic stem cell transplant, the method comprising administering to the human patient a ligand or fragment thereof that can bind to CD45 in an amount sufficient to deplete the population of CD45+ cells in the patient, and subsequently administering to the patient a graft comprising hematopoietic stem cells.

[0046]

[0047] In another aspect, the invention provides a method of depleting the population of CD34+ cells in a human patient by administering an effective amount of a ligand or fragment thereof that can bind to CD34. To provide the law.

[0046] In another embodiment, the present invention relates to the binding of CD34 to a patient before the patient receives a graft containing hematopoietic stem cells. By administering an effective amount of the ligand or fragment thereof, hematopoiesis can be achieved. This provides a method for depleting the CD34+ cell population in human patients requiring stem cell transplantation. .

[0047] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient A ligand or fragment that can bind to CD34 in a patient with CD34+ This method is characterized by pre-administering a sufficient amount to deplete a population of cells.

[0048] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method for providing a human patient with a ligand or fragment that can bind to CD34. The process involves administering the drug in an amount sufficient to deplete the population of CD34+ cells in the patient, and continuing... The method is characterized by the step of administering a graft containing hematopoietic stem cells to a patient.

[0049] In another aspect, the present invention relates to a ligand or fragment that can bind to CD90. By administering an effective dose, the method of depleting the population of CD90+ cells in human patients. To provide the law.

[0050] In another aspect, the present invention relates to the binding of CD90 to a graft containing hematopoietic stem cells before the patient receives the graft. By administering an effective amount of the ligand or fragment thereof, hematopoiesis can be achieved. This provides a method for depleting the CD90+ cell population in human patients requiring stem cell transplantation. .

[0051] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient A ligand or fragment that can bind to CD90, in patients with CD90+ This method is characterized by pre-administering a sufficient amount to deplete a population of cells.

[0052] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method for providing a human patient with a ligand or fragment that can bind to CD90. The process involves administering the drug in an amount sufficient to deplete the population of CD90+ cells in the patient, and continuing... The method is characterized by the step of administering a graft containing hematopoietic stem cells to a patient.

[0053] In another aspect, the present invention relates to a ligand or fragment that can bind to CD110. By administering an effective dose of the drug, the population of CD110+ cells in human patients is depleted. Provide a method.

[0054] In another embodiment, the present invention binds to CD110 before the patient receives a graft containing hematopoietic stem cells. By administering an effective amount of the ligand or fragment thereof, it is possible to create This invention provides a method to deplete the CD110+ cell population in human patients requiring hematopoietic stem cell transplantation. ru.

[0055] In another aspect, the present invention relates to, for example, a method for treating human patients who require hematopoietic stem cell transplantation. A law that includes the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient A ligand or fragment that can bind to CD110 in patients +A method characterized by pre-administering a sufficient amount to deplete a population of cells.

[0056] In a further embodiment, the present invention may, for example, treat human patients who require hematopoietic stem cell transplantation. A method for providing a human patient with a ligand or flag capable of binding to CD110. The process involves administering a dose of ment in a quantity sufficient to deplete the population of CD110+ cells in the patient, The method is characterized by the step of subsequently administering a graft containing hematopoietic stem cells to the patient.

[0057] In some embodiments of any of the above-described aspects, CD45 (e.g., CD45RO), CD34, CD A ligand or fragment that binds to one or more of 90 and CD110 is a human anti- A single human antibody isolated from the body (e.g., from IgG1, IgG2, IgG3, or IgG4 isotypes) It is covalently bonded to an Fc domain, such as a (separated) dimer Fc domain. Several embodiments So, the Fc domain is a monomeric Fc domain containing a single-stranded polypeptide chain. How many? In one embodiment, the N-terminus of the ligand or fragment is bound to the Fc domain. In some embodiments, the C-terminus of the ligand or its fragment is Fc-domain. It is bound to the ligand. The Fc domain is bound to one or more copies of the ligand or fragment. It may be conjugated to. For example, it may be used in conjunction with the methods described herein. The conjugate is formed when each polypeptide chain in the Fc domain is connected to the ligand or fragment. It contains a conjugated dimeric Fc domain. The Fc domain is described below. Cytotoxins (e.g., Pseudomonas exotoxin A, deBouganin, diphtheria toxin, α-flaxseed toxin) Amatoxins such as tin, saporins, maytansine, maytansinoids, auristatin Anthracyclines, calicheamicin, irinotecan, SN-38, duocalmycin Pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine Cytotoxins such as benzodiazepines (and indolinobenzodiazepine dimers, or their variants) It can also be conjugated.

[0058] In some embodiments of any of the aforementioned aspects, the ligand or its fragment is , cytotoxins as described herein (e.g., Pseudomonas exotoxin A, deBouganin, diphtheria Amatoxins such as α-amanitin, saporins, meitansine, meitansine D, auristatin, anthracycline, calicheamicin, irinotecan, SN-38, Duocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, India Rinobenzodiazepines, and indolinobenzodiazepine dimers, or their mutations It is covalently bound to cytotoxins such as those in the body. In some embodiments, the ligand or The N-terminus of the fragment is bound to a cytotoxin. In some embodiments, a ligand Alternatively, the C-terminus of the fragment is bound to a cytotoxin. The cytotoxin then binds to the Fc-dominant It can also be conjugated in.

[0059] In some embodiments of any of the aforementioned aspects, the ligand or its fragment is , one site of the ligand or its fragment (e.g., the ligand or its fragment) It is covalently bound to the cytotoxin at the N or C terminus of the ligand, and its fragment The Fc domain is located at another site (for example, the opposite end of the ligand or its fragment). It is covalently bonded to it.

[0060] In some embodiments of any of the above-described aspects, the Fc domain is a human IgG1 isotype. It is an Fc domain. In some embodiments, the Fc domain is a human IgG2 isotype Fc It is a domain. In some embodiments, the Fc domain is a human IgG3 isotype Fc domain. In some embodiments, the Fc domain is the human IgG4 isotype Fc domain. That is the case.

[0061] In some embodiments of the above-described models, the cytotoxin is α-amanitin, β-amanitin Manitin, γ-amanitin, ε-amanitin, amanin, amaninamide, amanulin, A Manulinic acid, and amatoxins such as proamanulin, or their derivatives. In some embodiments of the cytotoxin, the cytotoxin is amatoxin, and the cytotoxin is The dejugated antibody, its antigen-binding fragment, or ligand is represented by the formula Ab-Am. In the formula, Ab is an antibody, its antigen-binding fragment, or ligand, and Am is an amatoxyl. In some embodiments, Am is represented by formula (I): [ka] (I) [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, ORB or OR C and; R A and R B together with the oxygen atom to which they are attached form an optionally substituted 5-membered heterocycloalkyl group; formed; R3 is H, R C or R D and; R4 is H, OH, OR C OR D R C or R D and; R5 is H, OH, OR C OR D R C or R D and; R6 is H, OH, OR C OR D R C or R D and; R7 is H, OH, OR C OR D R C or R D and; R8 is OH, NH2, OR C OR D NHR C or NR C R D and; R9 is H, OH, OR C or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -L-Z; R D is optionally substituted alkyl (e.g., C1-C6 alkyl), optionally substituted hetero lower alkyl (e.g., C1-C6 heteroalkyl), optionally substituted alkenyl (e.g., C 2-C6 alkenyl), optionally substituted heteroalkenyl (e.g., C2-C6 heteroalkenyl (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; and to Z is a reactive substituent present on L, and CD45 (CD45RO, etc.), CD135, CD34, CD90, and Present in the antibody that binds to CD110, its antigen-binding fragment, or its ligand. It is a chemical site formed by a coupling reaction with a reactive substituent.

[0062] In some embodiments, Am is just one R C Includes substituents.

[0063] In some embodiments, Am is represented by formula (IA): [ka] (IA) [In the formula, R1 is H, OH, ORA , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4 is H, OH, OR C , OR D , R C , or R D and; R5 is H, OH, OR C , OR D , R C , or R D and; R6 is H, OH, OR C , OR D , R C , or R D and; R7 is H, OH, OR C , OR D , R C , or R D and; R8 is OH, NH2, OR C , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; Z is a reactive substituent present on L, and CD45 (CD45RO, etc.), CD135, CD34, CD90, and Present in the antibody that binds to CD110, its antigen-binding fragment, or its ligand. It is a chemical site formed by a coupling reaction with a reactive substituent; and In the formula, Am is just one R C [Contains substituents].

[0064] In some embodiments, Am is represented by formula (IB): [ka] (IB) [wherein, R1 is H, OH, OR A , or OR C ; R2 is H, OH, OR B , or OR C ; R A and R B together with the oxygen atom to which they are attached form an optionally substituted 5-membered heterocycloalkyl group; ; R3 is H, R C , or R D ; R4 is H, OH, OR C , OR D , R C , or R D ; R5 is H, OH, OR C , OR D , R C , or R D ; R6 is H, OH, OR C , OR D , R C , or R D ; R7 is H, OH, OR C , OR D , R C , or R D ; R8 is OH, NH2, OR C , OR D , NHR C , or NR C R D ; R9 is H, OH, OR C , or OR D ; X is -S-, -S(O)-, or -SO2-; R C is -L-Z; R D is optionally substituted alkyl (e.g., C1-C6 alkyl), optionally substituted hete C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; Z is a reactive substituent present on L, and CD45 (CD45RO, etc.), CD135, CD34, CD90, and Present in the antibody that binds to CD110, its antigen-binding fragment, or its ligand. It is a chemical site formed by a coupling reaction with a reactive substituent; and In the formula, Am is just one R C [Contains substituents].

[0065] In some embodiments, R A and R B They are together with the oxygen atoms to which they are bonded. They then combine to form the following: [ka] [In the formula, Y is O, S, NR E , and CR E R E Selected from, and R E and R E' Each of these is independently and arbitrarily substituted C1-C6 alkylene-R C , replace as desired C1-C6 heteroalkylene-R C , optionally substituted C2-C6 alkenylene-R C , replace as desired C2-C6 heteroalkenylene-R C , optionally substituted C2-C6 alkynylene-R C , place as you like Replaced C2-C6 heteroalkylene-R C , optionally substituted cycloalkylene-R C , at will Substituted heterocycloalkylene-R C , optionally substituted arylene-R C , or optionally Substituted heteroarylene-R C It is.

[0066] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] R3 is H or RC and; R4 is H, OH, OR C , OR D , R C , or R D and; R5 is H, OH, OR C , OR D , R C , or R D and; R6 is H, OH, OR C , OR D , R C , or R D and; R7 is H, OH, OR C , OR D , R C , or R D and; R8 is OH, NH2, OR C , or NHR C and; R9 is either H or OH; and In the formula, R C and R D These are defined as described above.

[0067] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] R3 is H or R C and; R4 and R5 are independently H, OH, and OR, respectively. C , R C , or OR Dand; R6 and R7 are H, respectively; R8 is OH, NH2, OR C , or NHR C and; R9 is either H or OH; and In the formula, R C This is as defined above.

[0068] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 is H, OH, or OR A and; R2 is H, OH, or OR B and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] R3, R4, R6, and R7 are each H; R5 is OR C and; R8 is either OH or NH2; R9 is either H or OH; and In the formula, R C This is as defined above.

[0069] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 and R2 are independently either H or OH; R3 is R C and; R4, R6, and R7 are H, respectively; R5 is H, OH, or OC1-C6 alkyl; R8 is either OH or NH2; R9 is either H or OH; and In the formula, R C This is as defined above.

[0070] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 and R2 are independently either H or OH; R3, R6, and R7 are each H; R4 and R5 are independently H, OH, and OR, respectively. C , or R C and; R8 is either OH or NH2; R9 is either H or OH; and In the formula, R C This is as defined above.

[0071] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 and R2 are independently either H or OH; R3, R6, and R7 are each H; R4 and R5 are independently either H or OH; R8 is OH, NH2, OR C , or NHR C and; R9 is either H or OH; and In the formula, R C This is as defined above.

[0072] In some embodiments, Am is represented by equation (II): [ka] [In the formula, X is S, SO, or SO2; R1 is H, or an antibody or its antigen-binding funnel] It is a linker covalently bonded to the ligment; and R2 is H, or antibody or so It is a linker covalently bonded to the antigen-binding fragment; where R1 is H R2 is a linker, and if R2 is H, then R1 is a linker.

[0073] In some embodiments of the above-described model, the cytotoxin is a group consisting of DM1 and DM4. A more selectable maytansinoid. In some embodiments, the cytotoxin is monomer A selection from the group consisting of chilu auristatin E and monomethyl auristatin F Statins are present. In some embodiments, the cytotoxin is daunorubicin, doxorubicin. Anthracyclines selected from the group consisting of epirubicin and idarubicin. ru.

[0074] In some embodiments of the above-described model, an antibody or its antigen-binding fragment This refers to monoclonal antibodies or their antigen-binding fragments, polyclonal antibodies or so Antigen-binding fragments, humanized antibodies or their antigen-binding fragments, bispecific antibodies or its antigen-binding fragment, bivariable immunoglobulin domain, single-stranded Fv molecule (sc Fv), diabody, triabody, nanobody, antibody-like protein scaffold, Fv fragment The following are selected from the group consisting of t, Fab fragment, F(ab')2 molecule, and tandem discFV. In some embodiments, the antibody is selected from the group consisting of IgG, IgA, IgM, IgD, and IgE. It has an isotype that is used.

[0075] In some embodiments of the above-described model, an antibody, its antigen-binding fragment, Alternatively, the ligand, after administration to the patient, can destroy hematopoietic cells such as hematopoietic stem cells, cancer cells, or autologous cells. They are taken up by immune cells. For example, antibodies, their antigen-binding fragments, or ligands via receptor-mediated endocytosis (e.g., cell surface CD45, CD135, CD34, CD90) (or when bound to CD110), it is taken up by hematopoietic stem cells, cancer cells, or autoimmune cells. It can be rare. In some embodiments, it is covalently bound to the antibody or its antigen-binding fragment. The cytotoxin is cleaved by chemical cleavage (for example, by the enzymatic or linker described herein). It can then be released into the cell (by nonspecific cleavage). The cytotoxin is then released, among other things, into the transplantation therapy. Endogenous hematopoietic cells such as endogenous hematopoietic stem cells, endogenous cancer cells, or endogenous autoimmune cells before treatment To promote cell death, its intracellular targets (in particular, the mitotic spindle apparatus, nuclear DNA, It can access ribosomal RNA (or topoisomerase, etc.).

[0076] In some embodiments of any of the above-described aspects, an antibody and its antigen-binding agent are used. Fragments, or ligands, can promote necrosis of hematopoietic stem cells. In this embodiment, the antibody or its antigen-binding fragment is administered to the patient in one or more cases. The above complement proteins, natural killer (NK) cells, macrophages, neutrophils, and / Alternatively, by supplementing hematopoietic stem cells with eosinophils, endogenous hematopoietic stem cells can be supplied before transplantation. It can accelerate death.

[0077] In some embodiments, the graft containing hematopoietic stem cells is an antibody or its antigen-binding flag. The ment is administered to the patient after its concentration has been substantially removed from the patient's blood.

[0078] In some embodiments, hematopoietic stem cells or their offspring are transplanted into a patient. From 2 days or more (for example, approximately 2-5 days, approximately 2-7 days, approximately 2-20 days, approximately 2-30 days, for example, 2 days) , 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 1 7th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th After days or more, the functional capacity of hematopoietic stem cells is maintained.

[0079] In some embodiments, hematopoietic stem cells or their offspring are localized in hematopoietic tissue such as bone marrow. This involves restoring hematopoiesis after transplanting hematopoietic stem cells into a patient. ru.

[0080] In some embodiments, during transplantation into a patient, hematopoietic stem cells are converted into megakaryocytes, platelets, and blood cells. Platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural killer cells, T phosphorus This induces the recovery of a cell population selected from a group consisting of pocytes and B lymphocytes.

[0081] In some embodiments of the above-described model, the method involves transplanted hematopoietic stem cells being To provide a niche where it may be possible to target the patient's hematopoietic stem cells before hematopoietic stem cell transplantation therapy. It is used to treat one or more disorders by depleting the population, etc. After transplantation, Hematopoietic stem cells are either deficient in the patient's cell type, or actively killed, or, for example, For example, productive hematopoiesis can be established to replenish cell types that have been killed by chemotherapy. For example, the patient may be a patient suffering from stem cell dysfunction. In some embodiments The patient had sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and The patient suffers from a hemoglobin disorder such as Iscott-Aldrich syndrome. , congenital immunodeficiency disorder or acquired immunodeficiency disorder (for example, human immunodeficiency virus or They may have an immunodeficiency disorder such as acquired immunodeficiency syndrome. In terms of treatment methods, patients have glycogen storage disorders, mucopolysaccharidosis, Gaucher disease, Haller disease, and sulfamethoxazole. They suffer from metabolic disorders such as ingolipidosis and metachromatic leukodystrophy. In some embodiments, patients have adenosine deaminase deficiency and severe combined immunodeficiency. Allergic hyperimmune globulin M syndrome, Chediak-Higashi syndrome, hereditary lymphohistiocytosis Osteopetrosis, osteogenesis imperfecta, saccharomyceta, thalassemia major, systemic sclerosis, systemic erythema The patient suffers from a disorder selected from the group consisting of thematosus and juvenile rheumatoid arthritis. In some embodiments, the patient had scleroderma, multiple sclerosis, ulcerative colitis, or Crohn's disease. , and suffering from autoimmune diseases such as type 1 diabetes. In some embodiments, patients They suffer from cancer such as blood cancer or myeloproliferative disorders. In some embodiments, The patients have acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, and chronic lymphoblastic leukemia. Hematological disease, multiple myeloma, diffuse large B-cell lymphoma, or non-Hodgkin lymphoma They are suffering from a myelodysplastic disorder such as myelodysplastic syndrome. They are suffering from the disease.

[0082] In some embodiments of the above-described models, this method is used for CD45+, CD135+, CD34+, and CD Cancer characterized by 90+ or ​​CD110+ cells (e.g., CD45+, CD135+, CD34+, CD90+, It is also used to directly treat cancers such as leukemia characterized by CD110+ cells. However, this refers to clusters of CD45+, CD135+, CD34+, CD90+, or CD110+ cancer cells in the patient. By administering antibodies that deplete the group, their antigen-binding fragments, or ligands, And / or to deplete the population of endogenous hematopoietic stem cells before hematopoietic stem cell transplantation, antibodies, This is done by administering an antigen-binding fragment or ligand. In the latter case, transplantation is performed, for example. Cancer can later rebuild the cell population that was depleted during the process of eradicating cancer cells. Myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, multiple bone Hematological cancers such as myeloma, diffuse large B-cell lymphoma, or non-Hodgkin lymphoma It is possible.

[0083] In some embodiments of the above-described model, the method is used to treat autoimmune diseases. It is used for purposes such as CD45+, CD135+, CD34+, CD90+, or CD110+ self-conducting. Antibodies, their antigen-binding fragments, or ligands are used to deplete a population of immune cells. By administering and / or before hematopoietic stem cell transplantation, the accumulation of endogenous hematopoietic stem cells By administering antibodies, their antigen-binding fragments, or ligands to deplete the group. In the latter case, transplantation is, for example, a population of cells that have been depleted during the process of eradicating autoimmune cells. It can be reconstructed later. Autoimmune diseases may include, for example: scleroderma, multiple sclerosis. (MS), Human Systemic Lupus (SLE), Rheumatoid Arthritis (RA), Inflammatory Bowel Disease (IBD), Treatment of Psoriasis Diabetes mellitus, type 1 diabetes mellitus, acute disseminated encephalomyelitis (ADEM), Addison's disease, generalized dysplasia Pilosis, ankylosing spondylitis, antiphospholipid syndrome (APS), aplastic anemia, autoimmune hemolysis sexual anemia, autoimmune hepatitis, autoimmune otitis interna (AIED), autoimmune lymphoproliferative syndrome ( ALPS), autoimmune oophoritis, Barlow's disease, Behçet's disease, bullous pemphigoid, cardiomyopathy, Sha Gas sickness, chronic fatigue immunodeficiency syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Crohn's disease , cicatricial pemphigoid, celiac disease-dermatitis herpetiformis, cold agglutinin disease, CREST syndrome, Dogaud's disease , discoid lupus, autonomic nervous system dysfunction, endometriosis, essential mixed cryoglobulinemia, fibrosis Myalgia - Fibromyalgia, Goodpasture syndrome, Graves' disease, Guillain-Barré syndrome (GBS) Hashimoto's thyroiditis, hidradenitis suppurativa, idiopathic and / or acute thrombocytopenic purpura, idiopathic Pulmonary fibrosis, IgA neuropathy, interstitial cystitis, juvenile arthritis, Kawasaki disease, lichen planus, Lyme disease, Meniere's disease, mixed connective tissue disease (MCTD), myasthenia gravis, neurogenic muscle tone, opsocorticoid Myoclonus syndrome (OMS), optic neuritis, oat thyroiditis, pemphigus vulgaris, malignant Anemia, polychondritis, polymyositis and dermatomyositis, primary biliary cirrhosis, polymyositis nodosa Pulsitis, polyglandular syndrome, polymyalgia rheumatica, primary agammaglobulinemia, Raynaud's phenomenon Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjögren's syndrome, steroids If-person syndrome, Takayasu's arteritis, temporal arteritis (also known as "giant cell arteritis") ), ulcerative colitis, uveitis, vasculitis, vitiligo, vulvodynia ("vulvovaginitis"), and Wegener's granulomatosis.

[0084] Therefore, in some embodiments of any of the above aspects, the present invention relates to sickle cell anemia. Anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiscott-Aldrich It is characterized by methods for treating abnormal hemoglobin disorders such as Tzi syndrome. Several implementation forms In this context, the present invention relates to congenital immunodeficiency disorders or acquired immunodeficiency disorders (for example, human immunodeficiency This method is characterized by its treatment of immunodeficiency disorders such as all viral infections or acquired immunodeficiency syndrome. In some embodiments, the present invention relates to glycogen storage disease, mucopolysaccharidosis, and gauche. Metabolic diseases such as Hurler's disease, sphingolipidosis, and metachromatic leukodystrophy. The present invention is characterized by a method for treating the disorder. In some embodiments, the present invention relates to adenosine dea Minase deficiency and severe combined immunodeficiency, hyperimmunoglobulin M syndrome, Chediac syndrome. Higashi syndrome, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, stolic disorders, thalassemia It consists of major rheumatoid arthritis, systemic sclerosis, systemic lupus erythematosus, and juvenile rheumatoid arthritis. The present invention is characterized by a method for treating a disorder selected from a group. In some embodiments, the present invention is characterized by a method for treating a disorder selected from a group. Autoimmune diseases such as scleroderma, multiple sclerosis, ulcerative colitis, Crohn's disease, and type 1 diabetes. The present invention is characterized by a method for treating diseases. In some embodiments, the present invention is used for blood cancers and other diseases. The present invention is characterized by a method for treating cancer or myeloproliferative disorders. In some embodiments, the present invention Akira has acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, and chronic lymphoblastic leukemia. , treatment of multiple myeloma, diffuse large B-cell lymphoma, or non-Hodgkin lymphoma The method is characterized by the following: In some embodiments, the patient has myelodysplastic syndrome or other myelodysplastic syndrome. The patient suffers from a plastic disease. In these embodiments, the method is used with CD45, CD135, CD34, and CD90. , or an antibody that binds to CD110, its antigen-binding fragment, or ligand, and / or hematopoietic stem cell grafts according to any of the above embodiments and examples of the present invention This may include the process of administering the medication.

[0085] Similarly, in some embodiments of any of the above aspects, the present invention relates to CD45+, CD135+, C Cancer characterized by D34+, CD90+, or CD110+ cells (e.g., CD45+, CD135+, CD34+ Methods to directly treat cancers such as leukemia characterized by CD90+ or ​​CD110+ cells. Provided. In these embodiments, the method is provided for CD45, CD135, CD34, CD90, or CD110. The process includes administering an antibody that binds to, its antigen-binding fragment, or a ligand. This includes acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, and chronic lymphoblastic leukemia. Hematologic conditions such as multiple myeloma, diffuse large B-cell lymphoma, or non-Hodgkin lymphoma. It could be fluid cancer.

[0086] Furthermore, in some embodiments of any of the above aspects, the present invention includes the following self Providing methods to treat immune diseases: multiple sclerosis (MS), human systemic lupus (SLE), Treatment of rheumatoid arthritis (RA), inflammatory bowel disease (IBD), psoriasis, type 1 diabetes mellitus, Acute disseminated encephalomyelitis (ADEM), Addison's disease, alopecia generalis, ankylosing spondylitis, antiphospholipid systemic syndrome (APS), aplastic anemia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmunity Otitis interna (AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmune oophoritis, Barlow Diseases, Behçet's disease, bullous pemphigoid, cardiomyopathy, Chagas disease, chronic fatigue immunodeficiency syndrome ( CFIDS), chronic inflammatory demyelinating polyneuropathy, Crohn's disease, pemphigoid scarring, celiac disease Rash dermatitis, cold agglutinin disease, CREST syndrome, Dogaud's disease, discoid lupus, autonomic imbalance, children Endometriosis, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, Goodpasture - syndrome, Graves' disease, Guillain-Barré syndrome (GBS), Hashimoto's thyroiditis, hidradenitis suppurativa, Idiopathic and / or acute thrombocytopenic purpura, idiopathic pulmonary fibrosis, IgA neuropathy, interstitial Cystitis, juvenile arthritis, Kawasaki disease, lichen planus, Lyme disease, Meniere's disease, mixed connective tissue disease (MCTD), Myasthenia Gravis, Neuromuscular Tonicity, Opsoclonus-Myoclonus Syndrome OMS, optic neuritis, oat thyroiditis, pemphigus vulgaris, pernicious anemia, polychondritis, polymyositis Inflammation and dermatomyositis, primary biliary cirrhosis, polyarteritis nodosa, polyglandular syndrome, rheumatic polymyositis Muscle pain, primary agammaglobulinemia, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, Sarcoidosis, scleroderma, Sjögren's syndrome, Stiffperson syndrome, Takayasu's arteritis , temporal arteritis (also known as "giant cell arteritis"), ulcerative colitis, uveitis , vasculitis, vitiligo, vulvodysia ("vulvar vestibular inflammation"), and Wegener's granulomatosis. These In this embodiment, the method involves an antibody that binds to CD45, CD135, CD34, CD90, or CD110, and The process includes administering an antigen-binding fragment or ligand.

[0087] In another embodiment, the present invention relates to a method for depleting a population of CD45+ cells, wherein the population is given the formula Ab- The method is characterized by contacting an effective amount of the conjugate represented by Am, where Ab This is an antibody or antigen-binding fragment that binds to CD45, and Am is an amatoxin. Am can be expressed by equation (IA): [ka] (IA) [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4, R5, R6, and R7 are independently H, OH, and OR, respectively. C , OR D , R C , or R D and ; R8 is OH, NH2, OR C , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; and to Z consists of a reactive substituent present on L and a substituent present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with a reactive substituent, In the formula, Am is just one R C [Contains substituents].

[0088] In some embodiments, Am is represented by formula (IB): [ka] (IB) [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4, R5, R6, and R7 are independently H, OH, and OR, respectively. C , OR D , R C , or R D and ; R8 is OH, NH2, OR C , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), an arbitrarily substituted hetero Roalkylenes (e.g., C1-C6 heteroalkylenes), optionally substituted alkenylenes (e.g.) For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 heteroalkenylenes) Roalkennyrenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene that has been arbitrarily substituted (e.g., C2-C6 heteroalkylene), arbitrarily substituted substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted Linkers such as arylene or optionally substituted heteroarylenes; and Z consists of a reactive substituent present on L and a substituent present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with a reactive substituent, In the formula, Am is just one R C [Contains substituents].

[0089] In another embodiment, the present invention is characterized by a conjugate represented by the formula Ab-Am, where Ab is It is an antibody or antigen-binding fragment that binds to CD45, and Am is an amatoxin. In some embodiments, Am is represented by the above formula (IA) or formula (IB).

[0090] In some embodiments of the two aforementioned aspects, the antibody or its antigen-binding fragment is via cysteine ​​residues in the Fc domain of the antibody or its antigen-binding fragment It is conjugated with xin. In some embodiments, the cysteine ​​residue is an antibody This is introduced by a mutation in the Fc domain of the antigen-binding fragment. The cysteine ​​residue can be selected from the group consisting of Cys118, Cys239, and Cys265.

[0091] In some embodiments of these models, the cysteine ​​residue binds to the antibody or its antigen. It is naturally present in the Fc domain of the fragment. For example, the Fc domain is found in human IgG1 Fc domain. It may be an IgG Fc domain such as cysteine, and the cysteine ​​residues are Cys261, Cys321, Cys367, and The group consisting of and can be selected from.

[0092] In some embodiments of these models, R1 is H, OH, or OR A and; R2 is H, OH, or OR B and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] ; R3, R4, R6, and R7 are each H; R5 is OR C and; R8 is either OH or NH2; and R9 is either H or OH.

[0093] In some embodiments, R1 and R2 are independently H or OH; R3 is R C and; R4, R6, and R7 are H, respectively; R5 is H, OH, or OC1-C6 alkyl; R8 is either OH or NH2; and R9 is either H or OH.

[0094] In some embodiments, R1 and R2 are independently H or OH; R3, R6, and R7 are each H; R4 is OR C , or R C and; R5 is H, OH, or OC1-C6 alkyl; R8 is either OH or NH2; and R9 is either H or OH.

[0095] In some embodiments, R1 and R2 are independently H or OH; R3, R6, and R7 are each H; R4 and R5 are independently either H or OH; R8 is OR C or NHR C and; as well R9 is either H or OH.

[0096] In some embodiments of these models, the antibody or its antigen-binding fragment is CD It is taken up by 45+ cells.

[0097] In some embodiments of these models, the antibody or its antigen-binding fragment is 1 Less than μM, less than 750 nM, less than 500 nM, less than 250 nM, less than 200 nM, less than 150 nM, less than 100 nM, 75 nM Less than 50 nM, less than 10 nM, less than 1 nM, less than 0.1 nM, less than 10 pM, less than 1 pM, or less than 0.1 pM K d It is coupled to CD45. In some embodiments, K d The concentration is approximately 0.1 pM to 1 μM. In one embodiment, K d It is approximately 357 pM, and k on It is approximately 2.56 × 10 5 M -1 s -1 and / Ta is K off It is approximately 9.14 x 10 -5 s -1 That is the case.

[0098] In some embodiments of these models, the antibody or its antigen-binding fragment is approximately 9×10 -2 M -1 s -1~Approx. 1×10 2 M -1 s -1 k on Then combine it with CD45.

[0099] In some embodiments of these models, the antibody or its antigen-binding fragment is CD The binding of 45 to the second antibody or its antigen-binding fragment is competitively inhibited, and here the second The antibody or its antigen-binding fragment has the following complementarity-determining region (CDR): a. CDR-H1 having the amino acid sequence SYAMS (SEQ ID NO: 16); b. CDR-H2 having the amino acid sequence AISGSGGSTFYADSVRG (SEQ ID NO: 17); c. CDR-H3 having the amino acid sequence EVMGPIFFDY (SEQ ID NO: 18); d. CDR-L1 having the amino acid sequence RASQSIISSALA (SEQ ID NO: 19); e. CDR-L2 having the amino acid sequence GASSRAT (SEQ ID NO: 20); and f. CDR-L3 having the amino acid sequence QQYGSTPLT (SEQ ID NO: 21).

[0100] In some embodiments of these models, the antibody or its antigen-binding fragment is It is YTH24.5 or its humanized variant.

[0101] In some embodiments of these models, the antibody or its antigen-binding fragment is CD The binding of 45 to the second antibody or its antigen-binding fragment is competitively inhibited, and here the second The antibody or its antigen-binding fragment is rat YTH24.5 or its humanized variant. .

[0102] In some embodiments of these models, the antibody or its antigen-binding fragment is mo Noclonal antibodies or their antigen-binding fragments, polyclonal antibodies or their antigens Binding fragment, humanized antibody or its antigen-binding fragment, bispecific antibody or The antigen-binding fragment, the bivariable immunoglobulin domain, the single-stranded Fv molecule (scFv), Diabody, triabody, nanobody, antibody-like protein scaffold, Fv fragment, Fa The following are selected from the group consisting of b fragments, F(ab')2 molecules, and tandemdi scFV.

[0103] In another embodiment, the present invention relates to a method for depleting a population of CD135+ cells, wherein the population is depleted by formula Ab The method is characterized by contacting an effective amount of the conjugate represented by -Am, where A b is an antibody or antigen-binding fragment that binds to CD135, and Am is an amatoxin. Yes, there is. Am can be expressed by equation (IA): [ka] (IA) [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4, R5, R6, and R7 are independently H, OH, and OR, respectively. C , OR D , R C , or R D and ; R8 is OH, NH2, ORC , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; and to Z consists of a reactive substituent present on L and a substituent present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with a reactive substituent, In the formula, Am is just one R C [Contains substituents].

[0104] In some embodiments, Am is represented by formula (IB): [ka] (IB) [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4, R5, R6, and R7 are independently H, OH, and OR, respectively. C , OR D , R C , or R D and ; R8 is OH, NH2, OR C , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), an arbitrarily substituted hetero Roalkylenes (e.g., C1-C6 heteroalkylenes), optionally substituted alkenylenes (e.g.) For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 heteroalkenylenes) Roalkennyrenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene that has been arbitrarily substituted (e.g., C2-C6 heteroalkylene), arbitrarily substituted substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted Linkers such as arylene or optionally substituted heteroarylenes; and Z consists of a reactive substituent present on L and a substituent present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with a reactive substituent, In the formula, Am is just one R C [Contains substituents].

[0105] In another embodiment, the present invention is characterized by a conjugate represented by the formula Ab-Am, where Ab is It is an antibody or antigen-binding fragment that binds to CD135, and Am is an amatoxin. In some embodiments, Am is represented by the above formula (IA) or formula (IB).

[0106] In some embodiments of the two aforementioned aspects, the antibody or its antigen-binding fragment is via cysteine ​​residues in the Fc domain of the antibody or its antigen-binding fragment It is conjugated with xin. In some embodiments, the cysteine ​​residue is an antibody This is introduced by a mutation in the Fc domain of the antigen-binding fragment. The cysteine ​​residue can be selected from the group consisting of Cys118, Cys239, and Cys265.

[0107] In some embodiments of these models, the cysteine ​​residue binds to the antibody or its antigen. It is naturally present in the Fc domain of the fragment. For example, the Fc domain is found in human IgG1 Fc domain. It may be an IgG Fc domain such as cysteine, and the cysteine ​​residues are Cys261, Cys321, Cys367, and The group consisting of and can be selected from.

[0108] In some embodiments of these models, R1 is H, OH, or OR A and; R2 is H, OH, or OR B and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] R3, R4, R6, and R7 are each H; R5 is OR C and; R8 is either OH or NH2; and R9 is either H or OH.

[0109] In some embodiments, R1 and R2 are independently H or OH; R3 is R C and; R4, R6, and R7 are H, respectively; R5 is H, OH, or OC1-C6 alkyl; R8 is either OH or NH2; and R9 is either H or OH.

[0110] In some embodiments, R1 and R2 are independently H or OH; R3, R6, and R7 are each H; R4 is OR C , or R C and; R5 is H, OH, or OC1-C6 alkyl; R8 is either OH or NH2; and R9 is either H or OH.

[0111] In some embodiments, R1 and R2 are independently H or OH; R3, R6, and R7 are each H; R4 and R5 are independently either H or OH; R8 is OR C or NHR C and; as well R9 is either H or OH.

[0112] In some embodiments of these models, the antibody or its antigen-binding fragment is CD It is taken up by 135+ cells.

[0113] In some embodiments of these models, the antibody or its antigen-binding fragment is 1 Less than μM, less than 750 nM, less than 500 nM, less than 250 nM, less than 200 nM, less than 150 nM, less than 100 nM, 75 nM Less than 50 nM, less than 10 nM, less than 1 nM, less than 0.1 nM, less than 10 pM, less than 1 pM, or less than 0.1 pM K d It is coupled to CD135. In some embodiments, K d The concentration is approximately 0.1 pM to 1 μM.

[0114] In some embodiments of these models, the antibody or its antigen-binding fragment is approximately 9×10 -2 M -1 s -1 ~Approx. 1×10 2 M -1 s -1 k on Then it is coupled to CD135.

[0115] In some embodiments of these models, the antibody or its antigen-binding fragment is The following CDRs are available: a. CDR-H1 having the amino acid sequence SYYMH (SEQ ID NO: 1); b. CDR-H2 having the amino acid sequence IINPSGGSTSYAQKFQG (SEQ ID NO: 2); c. CDR having amino acid sequence GVGAHDAFDI (SEQ ID NO: 3) or VVAAAVADY (SEQ ID NO: 4) -H3; d. Amino acid sequence RSSQSLLHSNGNNYLD (SEQ ID NO: 5) or RSSQSLLHSNGYNYLD (SEQ ID NO: 6) ) CDR-L1; e. CDR-L2 having the amino acid sequence LGSNRAS (SEQ ID NO: 7); and f. CDR- having amino acid sequence MQGTHPAIS (SEQ ID NO: 8) or MQSLQTPFT (SEQ ID NO: 9) L3.

[0116] In some embodiments of these models, the antibody or its antigen-binding fragment is The following CDRs are available: a. CDR-H1 having the amino acid sequence SYAIS (SEQ ID NO: 10); b. CDR-H2 having the amino acid sequence GIIPIFGTANYAQKFQG (SEQ ID NO: 11); c. CDR-H3 having the amino acid sequence FALFGFREQAFDI (SEQ ID NO: 12); d. CDR-L1 having the amino acid sequence RASQSISSYLN (SEQ ID NO: 13); e. CDR-L2 having the amino acid sequence AASSLQS (SEQ ID NO: 14); and f. CDR-L3 having the amino acid sequence QQSYSTPFT (SEQ ID NO: 15).

[0117] In some embodiments of these models, the antibody or its antigen-binding fragment is CD Competitively inhibits the binding of 135 to the second antibody or its antigen-binding fragment, where the second The antibody or its antigen-binding fragment has the following CDR: a. CDR-H1 having the amino acid sequence SYYMH (SEQ ID NO: 1); b. CDR-H2 having the amino acid sequence IINPSGGSTSYAQKFQG (SEQ ID NO: 2); c. CDR having amino acid sequence GVGAHDAFDI (SEQ ID NO: 3) or VVAAAVADY (SEQ ID NO: 4) -H3; d. Amino acid sequence RSSQSLLHSNGNNYLD (SEQ ID NO: 5) or RSSQSLLHSNGYNYLD (SEQ ID NO: 6) ) CDR-L1; e. CDR-L2 having the amino acid sequence LGSNRAS (SEQ ID NO: 7); and f. CDR- having amino acid sequence MQGTHPAIS (SEQ ID NO: 8) or MQSLQTPFT (SEQ ID NO: 9) L3.

[0118] In some embodiments of these models, the antibody or its antigen-binding fragment is CD Competitively inhibits the binding of 135 to the second antibody or its antigen-binding fragment, where the second The antibody or its antigen-binding fragment has the following CDR: a. CDR-H1 having the amino acid sequence SYAIS (SEQ ID NO: 10); b. CDR-H2 having the amino acid sequence GIIPIFGTANYAQKFQG (SEQ ID NO: 11); c. CDR-H3 having the amino acid sequence FALFGFREQAFDI (SEQ ID NO: 12); d. CDR-L1 having the amino acid sequence RASQSISSYLN (SEQ ID NO: 13); e. CDR-L2 having the amino acid sequence AASSLQS (SEQ ID NO: 14); and f. CDR-L3 having the amino acid sequence QQSYSTPFT (SEQ ID NO: 15).

[0119] In some embodiments of these models, the antibody or its antigen-binding fragment is mo Noclonal antibodies or their antigen-binding fragments, polyclonal antibodies or their antigens Binding fragment, humanized antibody or its antigen-binding fragment, bispecific antibody or The antigen-binding fragment, the bivariable immunoglobulin domain, the single-stranded Fv molecule (scFv), Diabody, triabody, nanobody, antibody-like protein scaffold, Fv fragment, Fa The following are selected from the group consisting of b fragments, F(ab')2 molecules, and tandemdi scFV.

[0120] In another embodiment, the present invention relates to a conjugate represented by the formula Ab-Cy, wherein Ab is C An antibody or antigen-binding fragment that binds to D45 (e.g., CD45RO), and in which Cy is finely divided. It is characterized by a conjugate that is a cytotoxin. In some embodiments of this model, it is a cytotoxin. The active ingredients are Pseudomonas exotoxin A, deBouganin, diphtheria toxin, saporin, and meitansine. Maytansinoids, auristatin, anthracyclines, calicheamicin, irino Tecan, SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolobenzodiazepine indolinobenzodiazepine dimer, or indolinobenzodiazepine dimer, Alternatively, it is a variant of one of the aforementioned cytotoxins.

[0121] In another embodiment, the present invention relates to a conjugate represented by the formula Ab-Cy, wherein Ab is C An antibody or antigen-binding fragment that binds to D135, and Cy is a cytotoxin. It is characterized by a cytotoxin. In some embodiments of this model, the cytotoxin is pseudomona Exotoxin A, deBouganin, diphtheria toxin, saporin, meitansine, meitansinoids Auristatin, anthracycline, calicheamicin, irinotecan, SN-38, Ocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indol Nobenzodiazepines, or indolinobenzodiazepine dimers, or the aforementioned cytotoxic It is a variant of one of the basic species.

[0122] In another embodiment, the present invention relates to a conjugate represented by the formula Ab-Cy, wherein Ab is C An antibody or antigen-binding fragment that binds to D34, and Cy is a cytotoxin. It is characterized by a cytotoxin. In some embodiments of this model, the cytotoxin is pseudomona Exotoxin A, deBouganin, diphtheria toxin, saporin, meitansine, meitansinoids Auristatin, anthracycline, calicheamicin, irinotecan, SN-38, Ocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indol Nobenzodiazepines, or indolinobenzodiazepine dimers, or the aforementioned cytotoxic It is a variant of one of the basic species.

[0123] In another embodiment, the present invention relates to a conjugate represented by the formula Ab-Cy, wherein Ab is C An antibody or antigen-binding fragment that binds to D90, and Cy is a cytotoxin. It is characterized by a cytotoxin. In some embodiments of this model, the cytotoxin is pseudomona Exotoxin A, deBouganin, diphtheria toxin, saporin, meitansine, meitansinoids Auristatin, anthracycline, calicheamicin, irinotecan, SN-38, Ocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indol Nobenzodiazepines, or indolinobenzodiazepine dimers, or the aforementioned cytotoxic It is a variant of one of the basic species.

[0124] In another embodiment, the present invention relates to a conjugate represented by the formula Ab-Cy, wherein Ab is C An antibody or antigen-binding fragment that binds to D110, and Cy is a cytotoxin. It is characterized by a cytotoxin. In some embodiments of this model, the cytotoxin is pseudomona Exotoxin A, deBouganin, diphtheria toxin, saporin, meitansine, meitansinoids Auristatin, anthracycline, calicheamicin, irinotecan, SN-38, Ocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indol Nobenzodiazepines, or indolinobenzodiazepine dimers, or the aforementioned cytotoxic It is a variant of one of the basic species.

[0125] In some embodiments of the five aspects described above, an antibody or its antigen-binding fragment hematopoietic stem cells (e.g., CD45+ (CD45RO+, etc.), CD135+, CD34+, CD90+, and / or It is taken up by hematopoietic cells such as CD110+ cells.

[0126] In some embodiments of these models, the antibody or its antigen-binding fragment is 1 Less than μM, less than 750 nM, less than 500 nM, less than 250 nM, less than 200 nM, less than 150 nM, less than 100 nM, 75 nM Less than 50 nM, less than 10 nM, less than 1 nM, less than 0.1 nM, less than 10 pM, less than 1 pM, or less than 0.1 pM. Full K d It combines with CD45 (e.g., CD45RO), CD135, CD34, CD90, or CD110. In that embodiment, K d The concentration is approximately 0.1 pM to 1 μM.

[0127] In some embodiments of this model, the antibody or its antigen-binding fragment is approximately 9 × 1 0 -2 M -1 s -1 ~Approx. 1×10 2 M -1 s -1 k on For example, CD45 (e.g., CD45RO), CD135, CD34, CD90, and It connects to CD110.

[0128] In some embodiments of these models, the antibody or its antigen-binding fragment is mo Noclonal antibodies or their antigen-binding fragments, polyclonal antibodies or their antigens Binding fragment, humanized antibody or its antigen-binding fragment, bispecific antibody or The antigen-binding fragment, the bivariable immunoglobulin domain, the single-stranded Fv molecule (scFv), Diabody, triabody, nanobody, antibody-like protein scaffold, Fv fragment, Fa Selected from the group consisting of b fragments, F(ab')2 molecules, and tandem di scFV. How many In that embodiment, the antibody is selected from the group consisting of IgG, IgA, IgM, IgD, and IgE. It has an isotype. [Brief explanation of the drawing]

[0129] [Figure 1] This graph shows the effect on the in vitro viability of Reh cells of various concentrations of anti-CD45 monoclonal antibodies Ab1, Ab2, and Ab3, each conjugated to a saporin via a saporin-conjugate Fab fragment, or a negative control with a matched isotype. Cell viability was evaluated using the CellTiter-Glo® assay kit, as described in Example 8 below. [Figure 2] This graph shows the effect on the in vitro viability of human CD34+ cells of various concentrations of anti-CD45 monoclonal antibodies or isotype-matched negative controls, each conjugated to a saporin via a saporin-conjugate Fab fragment. Cell viability was evaluated using the CellTiter-Glo® assay kit, as described in Example 8 below. [Modes for carrying out the invention]

[0130] This invention addresses a variety of disorders, particularly cell type diseases in the hematopoietic system, cancer, and autoimmunity. This specification provides methods for treating infectious diseases, metabolic disorders, and stem cell disorders, etc. The composition and method include (i) a population of cells that cause a disease, such as cancer cells (e.g., white By directly depleting populations of blood cells (e.g., autoimmune cells) and autoreactive T cells. (ii) obtain and / or (ii) engraftment of transplanted hematopoietic stem cells, and the transplanted cells are homing Depleting the population of endogenous hematopoietic stem cells by providing a niche where it can be promoted It is possible to cause this. The aforementioned activity is generated by cells or hematopoietic stem cells that cause endogenous diseases. An antibody, its antigen-binding fragment, or ligand capable of binding to an expressed antigen. This can be achieved by administering [the substance]. In the case of direct treatment of the disease, this administration is of interest. It can cause a decrease in the number of cells that cause certain disease conditions. If administered to a person, this administration may cause selective depletion of the endogenous hematopoietic stem cell population. This can occur, causing a vacancy in hematopoietic tissues such as bone marrow, which can later be transplanted. It can be supplemented with exogenous hematopoietic stem cells. The present invention relates to CD45, CD135, CD34, CD90, or C An antibody capable of binding to D110, its antigen-binding fragment, and ligand are provided to the patient. This is partly based on the discovery that both of the above activities can be achieved by administering it. Antibodies that bind to 45, CD135, CD34, CD90, or CD110, their antigen-binding fragments, and The ligand is administered to patients with cancer or autoimmune disease, and cancer cells or It can directly deplete a population of autoimmune cells, as well as the survival of transplanted hematopoietic stem cells. And to promote the chances of engraftment, it is administered to patients requiring hematopoietic stem cell transplantation. It is also possible to do so.

[0131] Anti-CD45 antibody, anti-CD135 antibody, anti-CD34 antibody, anti-CD90 antibody, or anti-CD110 antibody, and their anti- The engraftment of hematopoietic stem cell grafts by administration of primordial fragments or ligands varies. This can be revealed by empirical measurements. For example, the engraftment of transplanted hematopoietic stem cells is related to CD. Antibodies or antigen-binding agents capable of binding to 45, CD135, CD34, CD90, or CD110 After administration of lagment and subsequent administration of hematopoietic stem cell grafts, the presence of these cells in the patient's bone marrow This can be evaluated by measuring the amount of competitive reconstitution units (CRUs). Furthermore, fluorescence Reporter genes, such as enzymes that catalyze chemical reactions that produce color or light-emitting products, are used in Dona - Hematopoietic stem cells are incorporated into a transfected vector, followed by the corresponding sigma By monitoring the NARS in tissues where hematopoietic stem cells are homing, such as bone marrow, This allows us to observe the engraftment of hematopoietic stem cell grafts. For example, in the field of this technology As determined by known fluorescence-activated cell classification (FACS) analysis methods, hematopoietic stem cells Engraftment of hematopoietic stem cell grafts is observed by evaluating the quantity and survival of cells and progenitor cells. It can also be done by measuring the number of white blood cells in the peripheral blood during the post-transplant period. By measuring the recovery of bone marrow cells by donor cells in bone marrow aspirate samples, It can also be decided.

[0132] The following sections apply to patients with cancer or autoimmune diseases, or those undergoing hematopoietic stem cell transplantation. To administer to patients who require it, such as those who need it, in order to promote the engraftment of hematopoietic stem cells. Description of antibodies that can be produced, their antigen-binding fragments, and ligands, as well as such The present invention provides a method for administering a therapeutic drug to a patient (for example, before hematopoietic stem cell transplantation).

[0133] (definition) As used herein, the term "about" refers to a value within 10% above or below the stated value. For example, the term "approximately 5 nM" refers to a range of 4.5 nM to 5.5 nM.

[0134] As used herein, the term “amatoxin” refers to the production of Amanita phalloides by Amanita virosa. A member of the amatoxin family of peptides, or its variant or derivative. For example, a variant or derivative that can inhibit RNA polymerase II activity. Amatoxins useful in combination with the compositions and methods described herein include those with the following formula ( III) Compounds, for example, α-amanitin, β-amanitin, γ-amanitin, ε-amanitin Amanin, amaninamide, amanulin, amanuric acid, and proamanulin, etc. The following is an example. Equation (III) is as follows: [ka] [In the formula, R1 is H, OH, or OR A and; R2 is H, OH, or OR B and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H or R D and; R4 is H, OH, OR D , or R D and; R5 is H, OH, OR D , or R D and; R6 is H, OH, OR D , or R D and; R7 is H, OH, OR D , or R D and; R8 is OH, NH2, or ORD and; R9 is H, OH, or OR D and; X is -S-, -S(O)-, or -SO2-; and R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl compound.

[0135] For example, amatoxins useful in combination with the compositions and methods described herein include the following: Examples of compounds of formula (IIIA) include: [ka] [In the formula, R1 is H, OH, or OR A and; R2 is H, OH, or OR B and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H or R D and; R4 is H, OH, OR D , or R D and; R5 is H, OH, OR D , or R D and; R6 is H, OH, OR D , or R D and; R7 is H, OH, OR D , or R D and; R8 is OH, NH2, or OR D and; R9 is H, OH, or OR D and; X is -S-, -S(O)-, or -SO2-; and R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl compound.

[0136] Amatoxins useful in combination with the compositions and methods described herein include those of the following formula (II Compounds of type IB are also listed: [ka] [In the formula, R1 is H, OH, or OR A and; R2 is H, OH, or OR B and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H or R D and; R4 is H, OH, OR D , or R D and; R5 is H, OH, OR D , or R D and; R6 is H, OH, OR D , or R D and; R7 is H, OH, OR D , or R D and; R8 is OH, NH2, or OR D and; R9 is H, OH, or OR D and; X is -S-, -S(O)-, or -SO2-; and R D This includes optionally substituted alkyls (e.g., C1-C6 alkyls) and optionally substituted heteros. Alkyl (e.g., C1-C6 heteroalkyl), optionally substituted alkenyl (e.g., C2- C6 alkenyls, optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) ), arbitrarily substituted alkynyls (e.g., C2-C6 alkynyls), arbitrarily substituted hetyl Roalkynyl (e.g., C2-C6 heteroalkynyl), optionally substituted cycloalkyl, Optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted It is a heteroaryl compound.

[0137] As described herein, amatoxin can, for example, transmit antibodies via a linker site. amatoxin can be conjugated to its antigen-binding fragment or ligand. Exemplary methods of conjugation and linkers useful in such processes are described as "conjugation The section titled "Linker for Scientific Conjugation" and the following Table 2 are described below. The compositions and methods described herein are used to produce antibodies, antigen-binding fragments, or other substances. Exemplary linker-containing amatoxins useful for conjugation to Gand are described herein. This is shown in the structural formulas (I), (IA), (IB), and (II) listed below.

[0138] As used herein, the term “antibody” means an antibody that specifically binds to a particular antigen, or This refers to immunoglobulin molecules that are immunologically reactive with it, such as polyclonal and monoclonal immunoglobulin molecules. Examples include, but are not limited to, natural, genetically modified, and other modified forms of antibodies. , chimeric antibodies, humanized antibodies, heteroconjugate antibodies (e.g., double, triple, and quadruple conjugate antibodies) Specific antibodies, diabodies, triabodies, and tetrabodies), as well as, for example, Antigen binding of antibodies containing Fab', F(ab')2, Fab, Fv, rIgG, and scFV fragments. Fragments are mentioned. Unless otherwise specified, the term "monoclonal antibody" (mAb) is used. This involves intact molecules that can specifically bind to target proteins, and antibody fragments. This means including both the 'F' and 'F(ab')2' fragments. When used herein, Fab and F(ab')2 fragments are the Fc fragments of the intact antibody. This refers to antibody fragments lacking fragmentation. Examples of these antibody fragments are given herein. It is described there.

[0139] When used herein, the term “antigen-binding fragment” refers to a fragment that is specific to a target antigen. This refers to one or more fragments of an antibody that retain the ability to bind. The function can be performed by a fragment of a full-length antibody. The antibody fragment is, for example, Fa b, F(ab')2, scFV, Diabody, Triabody, Affibody, Nanobody, Aptama —or it may be a domain antibody. It is included in the term "antigen-binding fragment" of an antibody. Examples of combined fragments include, but are not limited to: (i)Fab Fragment, V L , V H , C L , and C H (ii) F(ab' )2 fragments, two Fab fragments linked by disulfide bridges in the hinge region (iii)V H and C H Fd fragment consisting of one domain; (iv ) Single-arm V of the antibody L and V H Fv fragment consisting of domains, (v)V H and V L Do dAb;(vi)V including the main H dAb fragments consisting of domains (e.g., Ward et al., N See ature 341:544-546, 1989; (vii)V H or V L dAb consisting of a domain; (viii) Isolated complementarity-determining regions (CDRs); (ix) optionally linked by a composite linker A good combination of two or more (e.g., 2, 3, 4, 5, or 6) isolated CDRs. The two domains of the Fv fragment, V L and V H These are encoded by separate genes. However, they use recombination methods, V L and V HThe regions pair up to form a monovalent molecule. Linkers enable them to be synthesized as a single protein chain. It can be bonded (known as single-stranded Fv (scFv); e.g., Bird et al., Science 242:423) See -426, 1988 and Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883, 1988. (See image). These antibody fragments can be obtained using prior art known to those skilled in the art. The fragments can be screened for usefulness in the same manner as intact antibodies. Antigen-binding fragments can be created using recombinant DNA technology, intact immunoglobulins. By enzymatic or chemical cleavage of brin, or in some cases known in the art It can be produced by established chemical peptide synthesis procedures.

[0140] As used herein, the term “anti-CD45 antibody” is specific to CD45 (e.g., CD45RO). At least a portion of an immunoglobulin molecule that can bind to, for example, not limited to However, at least one complementarity determination region of the heavy chain or light chain or their ligand binding sites Region (CDR), heavy or light chain variable region, heavy or light chain steady region, framework region This refers to a protein or peptide-containing molecule that includes a region, or any part thereof. CD45 antibodies have BC, DE, and FG structures that are similar in structure and solvent accessibility to antibody CDR. The 10th fibronectin type III domain containing the -p 10 Antibody-like proteins such as Fn3 The venue can also be mentioned. 10 The tertiary structure of the Fn3 domain is similar to that of the variable region of the IgG heavy chain. Those skilled in the art, for example,10 The residues of the BC, DE, and FG loops of Fn3 are anti-CD45 monoclonal By substituting residues from the CDRH-1, CDRH-2, or CDRH-3 region of the antibody, anti-CD45 mo CDRs of noclonal antibodies can be transplanted onto a fibronectin scaffold.

[0141] As used herein, the term "anti-CD135 antibody" means an antibody that specifically binds to CD135. It is possible to modify at least a portion of an immunoglobulin molecule, for example, but not limited to heavy chains or At least one complementarity-determining region (CDR) of the light chain or its ligand-binding site, and also of the heavy chain. or light chain variable region, heavy chain or light chain steady region, framework region, or a combination of these This refers to a protein or peptide-containing molecule that includes any site. Anti-CD135 antibodies include anti The tenth structure includes BC, DE, and FG structural loops that are similar in structure and solvent accessibility to the CDR. Fibronectin type III domain ( 10 Antibody-like protein scaffolds such as Fn3 are also examples. 10 The tertiary structure of the Fn3 domain is similar to that of the variable region of the IgG heavy chain, and those skilled in the art will see, for example, Ba, 10 The residues of the BC, DE, and FG loops of Fn3 were removed by the anti-CD135 monoclonal antibody CDRH-1, CD By substituting residues from the RH-2 or CDRH-3 region, anti-CD135 monoclonal anti Body CDRs can be transplanted onto a fibronectin scaffold.

[0142] As used herein, the term "anti-CD34 antibody" means an antibody that can specifically bind to CD34. This includes at least a portion of immunoglobulin molecules, for example, heavy chains or light chains, but are not limited to these. At least one complementarity-determining region (CDR) of the chain or its ligand-binding site, heavy chain also or light chain variable region, heavy chain or light chain steady region, framework region, or any of the above. This refers to a protein or peptide-containing molecule that includes the desired site, etc. Anti-CD34 antibodies include antibody CD The tenth f includes BC, DE, and FG structural loops that are similar in structure and solvent accessibility to R. Ibronectin type III domain ( 10 Antibody-like protein scaffolds such as Fn3 are also examples. 10 Fn The tertiary structure of the three domains is similar to the tertiary structure of the variable region of the IgG heavy chain, and those skilled in the art will know, for example, 10 The residues of the BC, DE, and FG loops of Fn3 are replaced by the anti-CD34 monoclonal antibodies CDRH-1 and CDRH-2. or by substituting residues from the CDRH-3 region, the CD34 monoclonal antibody R can be transplanted onto a fibronectin scaffold.

[0143] As used herein, the term “anti-CD90 antibody” means an antibody that can specifically bind to CD90. This includes at least a portion of immunoglobulin molecules, for example, heavy chains or light chains, but are not limited to these. At least one complementarity-determining region (CDR) of the chain or its ligand-binding site, heavy chain also or light chain variable region, heavy chain or light chain steady region, framework region, or any of the above. This refers to a protein or peptide-containing molecule that includes the desired site, etc. Anti-CD90 antibodies include antibody CD The tenth f includes BC, DE, and FG structural loops that are similar in structure and solvent accessibility to R. Ibronectin type III domain ( 10 Antibody-like protein scaffolds such as Fn3 are also examples. 10 Fn The tertiary structure of the three domains is similar to the tertiary structure of the variable region of the IgG heavy chain, and those skilled in the art will know, for example, 10 The residues of the BC, DE, and FG loops of Fn3 are replaced by the anti-CD90 monoclonal antibodies CDRH-1 and CDRH-2. or by substituting residues from the CDRH-3 region, the CD90 monoclonal antibody R can be transplanted onto a fibronectin scaffold.

[0144] As used herein, the term “anti-CD110 antibody” means an antibody that specifically binds to CD110. It is possible to modify at least a portion of an immunoglobulin molecule, for example, but not limited to heavy chains or At least one complementarity-determining region (CDR) of the light chain or its ligand-binding site, and also of the heavy chain. or light chain variable region, heavy chain or light chain steady region, framework region, or a combination of these This refers to a protein or peptide-containing molecule that includes any site. Anti-CD110 antibodies include anti- The tenth structure includes BC, DE, and FG structural loops that are similar in structure and solvent accessibility to the CDR. Fibronectin type III domain ( 10 Antibody-like protein scaffolds such as Fn3 are also examples. 10 The tertiary structure of the Fn3 domain is similar to that of the variable region of the IgG heavy chain, and those skilled in the art will see, for example, Ba, 10 The residues of the BC, DE, and FG loops of Fn3 were removed by the anti-CD110 monoclonal antibody CDRH-1, CD By substituting residues from the RH-2 or CDRH-3 region, anti-CD110 monoclonal anti Body CDRs can be transplanted onto a fibronectin scaffold.

[0145] As used herein, the term “bispecific antibody” means, for example, an antibody that has at least two different properties. Monoclonal antibodies capable of binding to a specific antigen, often human antibodies or humanized antibodies. This refers to antibodies. For example, one of the binding specificities could be CD45 (e.g., CD45RO), CD135, CD34, CD9 It can be directed to hematopoietic stem cell surface antigens such as 0 or CD110, and the other can be directed to different hematopoietic stem cells. Cell surface antigens or other cell surface proteins, among other things, signaling pathways that enhance cell proliferation. It can be directed towards receptors or receptor subunits involved in the pathway.

[0146] As used herein, the term “complementarity-determining region” (CDR) refers to the light chain variable domain of an antibody. This refers to the hypervariable region found in both the yin and heavy chain variable domains. The portion that is preserved each time is called the framework region (FR). The hypervariable region of the antibody is demarcated. The amino acid positions that are used depend on the context and the various definitions known in the relevant field. It can change. Some positions within a variable domain are transcendent under certain criteria. It can be considered to be within the variable region, but under different criteria it can be considered outside the hypervariable region. In that it can do so, it can be considered a hybrid ultra-variable position. One or more of these can also be seen in the extended hypervariable region. The antibodies described herein are this These may include modifications to the hybrid hypervariable position. The variable domains of the natural heavy and light chains are Each of these primarily consists of four framework areas with a β-sheet structure, and is comprised of three CDRs. They are connected, and they form loops that connect the β-sheet structure, and in some cases the β-sheet structure It forms part of the chain. The CDRs in each chain are in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. They are held together in close proximity by the muwer region, and together with CDR from other antibody chains, anti It contributes to the formation of target binding sites in the body (Kabat et al., Sequences of Proteins of Immunol (See *Optical Interest*, National Institute of Health, Bethesda, MD., 1987). When used in detailed documents, immunoglobulin amino acid residue numbering is as follows, unless otherwise specified. This is done according to the immunoglobulin amino acid residue numbering system developed by Kabat et al.

[0147] As used herein, the terms “conditioning” and “conditioning” "Preparing" refers to the process of preparing a patient to receive a graft containing hematopoietic stem cells. Procedures such as those described above promote the engraftment of hematopoietic stem cells (for example, conditioning procedures and And viable hematopoietic stem cells in blood samples isolated from patients after subsequent hematopoietic stem cell transplantation. (As can be inferred from the sustained increase in the number of cells). The method described herein involves hematopoietic stem cells Therefore, antigens expressed include CD45 (e.g., CD45RO), CD135, CD34, CD90, or CD110. By administering an antibody or its antigen-binding fragment that can bind to the patient, Patients can be prepared for hematopoietic stem cell transplantation therapy, as described herein. Antibodies covalently bind to cytotoxins, forming drug-antibody conjugates. It can bind to one or more of the aforementioned antigens. Antibodies that can bind to one or more of the aforementioned antigens, and their anti- Hematopoietic stem cell transplantation therapy with primordial-bound fragments or drug-antibody conjugates is required. Administration to patients who undergo treatment selectively depletes endogenous hematopoietic stem cells, thereby reducing exogenous hematopoietic stem cells. By creating a vacancy that is filled by hematopoietic stem cell transplantation, the life of the hematopoietic stem cell graft It can promote wearing.

[0148] As used herein, the term “conjugate” refers to an antibody or its antigen-binding flux. A reactive functional group of one molecule, such as a ligment, and another molecule, such as a cytotoxin as described herein. A conjugate refers to a compound formed by chemical bonding with an appropriate reactive functional group. It may contain a linker between two molecules that are bonded to each other. It can be used to form conjugates. Examples of linkers include naturally occurring amino acids or non-naturally occurring amino acids such as D-amino acids. Examples include peptide-containing linkers, such as those containing amino acids. The linker is described in this specification. It can be prepared using various strategies described in the book and known in the art. Depending on the reactive components within it, the linker can undergo, for example, enzymatic hydrolysis, photolysis, or acidic conditions. Hydrolysis under basic conditions, hydrolysis under basic conditions, oxidation, disulfide reduction, nucleophilic cleavage, and It can be cleaved by organometallic cleavage (e.g., Leriche et al., Bioorg. Med. Chem., See 20:571-582, 2012.

[0149] As used herein, the term "coupling reaction" refers to a reaction between two people that are suitable for each other to react. Two or more substituents react to form molecular fragments bonded to each substituent (for example, covalent bonds). This refers to a chemical reaction that forms chemical sites that (collectively) link together. Coupling reactions include cytotoxic reactions. Flavours, such as cytotoxins known in the art or described herein. The reactive substituent attached to the ligand is an antibody, its antigen-binding fragment, or ligand. For example, CD45 (such as CD45RO), which is known in the art or described herein. Antibodies specific to CD135, CD34, CD90, and / or CD110, their antigen-binding fragments, Alternatively, a ligand or other appropriate substituent that is bound to a fragment and reacts with it. Examples include appropriately reactive substituents, such as nucleophile / electrophile pairs (for example, bird Thiol / haloalkyl pair, amine / carbonyl pair, or thiol / α,β-unsaturated Carbonyl pairs, diene / dienofil pairs (for example, azid / alkyne pairs in particular), etc. Examples include coupling reactions, although not particularly limited, such as thiol alkylation and hydroxylation. Roxyl alkylation, amine alkylation, amine condensation, amidation, esterification, disulfide Addition formation, cycloaddition (for example, in particular, [4+2]Diels-Alder cycloaddition, [3+2 (Hüsgen cycloaddition), nucleophilic aromatic substitution, electrophilic aromatic substitution, and known in the art This includes the reaction described herein or other reaction methods described herein.

[0150] As used herein, "CRU (Competitive Reconstitution Unit)" refers to a unit that can be detected after in vivo transplantation. This refers to a unit of measurement for long-term engrafted stem cells.

[0151] As used herein, the term “donor” means one or more cells that are either those cells or other cells. Refers to a human or animal isolated before administration of its offspring to a recipient. One or more cells. This could be, for example, a population of hematopoietic stem cells.

[0152] As used herein, the term “diabody” refers to a structure containing two polypeptide chains. This refers to a bivalent antibody, where each polypeptide chain is on the same peptide chain. H and V L Domain molecule Linkers that are too short to allow internal association (for example, linkers consisting of 5 amino acids) V connected by H and V L It includes a domain. This configuration forms a homodimer structure. To that end, each domain is paired with a complementary domain on another polypeptide chain. The term "tribody" refers to a trivalent antibody containing three peptide chains, each of which is... V within the same peptide chain H Domain and V L To enable intramolecular association of domains, very short ri A single V linked by a linker (for example, a linker consisting of 1-2 amino acids) H Dome In and one V L It includes domains. It is constructed in this way in order to fold them into their natural structure. The peptides are typically V of the adjacent peptide chain. H and V L Domains in relation to each other Trimerization is performed to position the molecule proximal to the target (e.g., Holliger et al., Proc. Natl. Acad.). See Sci. USA 90:6444-48, 1993.

[0153] As used herein, “Dual Variable Domain Immunoglobulin” (“DVD-Ig”) is, The target-binding variable domains of two monoclonal antibodies are combined via a linker to form a tetravalent duplex antibody. This refers to antibodies used to produce single targeted agents (e.g., Gu et al., Meth. Enzymol., 502:25-41, 20 See 12).

[0154] As used herein, the term “endogenous” means that a particular organism, such as a human patient, is naturally occurring. Substances such as molecules, cells, tissues, or organs that can be affected (e.g., hematopoietic stem cells or hematopoietic lineage) Cells, such as megakaryocytes, platelets, red blood cells, mast cells, myeloblasts, basophils, and neutrophils. Eosinophils, microglia, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells This refers to cells (such as natural killer cells, T lymphocytes, or B lymphocytes).

[0155] As used herein, the term “engraftment potential” refers to the tissue of hematopoietic stem cells and progenitor cells. It is used to refer to the ability to reconstruct cells, and whether such cells circulate naturally or Regardless of whether they were supplied by transplantation. This term refers to the tissue homing and eyes of cells. All processes surrounding or leading up to engraftment, such as the formation of cell colonies within the target tissue. This includes the event. Engraftment efficiency or engraftment rate is any clinically acceptable rate known to those skilled in the art. The parameters used can be evaluated or quantified, for example, competitive restructuring. Evaluation of CRU (Cell Unit) status; tissue in which stem cells have homed, colonized, or engrafted (single unit) This may include the incorporation or expression of markers (or multiple markers); or the progression of the disease. Progression of the subject due to the survival of hematopoietic stem cells and progenitor cells, or the survival of the recipient. Engraftment is determined by evaluation. Engraftment is determined by measuring the white blood cell count in the peripheral blood during the post-transplant period. It is also possible to measure the recovery of bone marrow cells by donor cells in a bone marrow aspirate sample. It can also be evaluated by doing so.

[0156] As used herein, the term “exogenous” means that a particular organism, such as a human patient, does not naturally exist in it. Substances such as molecules, cells, tissues, or organs that are not found (e.g., hematopoietic stem cells or hematopoietic system) Cells in rows, such as megakaryocytes, platelets, erythrocytes, mast cells, myeloblasts, basophils, tetraphosphorus Mesoglobulins, eosinophils, microglial cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, trees This refers to exogenous substances (such as cytoplasmic cells, natural killer cells, T lymphocytes, or B lymphocytes). This includes things supplied from external sources to organisms, or to cultures extracted from them. It can be done.

[0157] As used herein, the term “framework region,” or “FW region,” refers to an antibody or containing amino acid residues adjacent to the CDR of the antigen-binding fragment. FW region residues Examples include, for instance, human antibodies, humanized antibodies, monoclonal antibodies, and antibody fragments. Fab fragment, single-chain antibody fragment, scFV fragment, antibody domain, and It can be present in bispecific antibodies.

[0158] As used herein, the term “hematopoietic stem cell” (“HSC”) means a self-replicating and the following: Immature blood cells that have the ability to differentiate into mature blood cells, including, but not limited to, a diverse range of lineages. Refers to blood cells: granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), red blood cells (e.g., Reticulocytes, red blood cells), platelets (e.g., megakaryocytes, platelet-producing megakaryocytes, platelets), monocytes ( For example, monocytes, macrophages, dendritic cells, microglia, osteoclasts, and lymphocytes (e.g.) For example, NK cells, B cells, and T cells. Such cells could include CD34+ cells. D34+ cells are immature cells that express the CD34 cell surface marker. In humans, CD34+ cells are It is thought to include a subpopulation of cells having the stem cell characteristics defined above, but in mice, HS C is CD34-. Furthermore, HSC is divided into Long-Term Reconstruction HSC (LT-HSC) and Short-Term Reconstruction HSC (ST-HSC). C) also refers to LT-HSCs and ST-HSCs based on their functional capacity and cell surface marker expression. They are distinguished. For example, human HSCs are CD34+, CD38-, CD45RA-, CD90+, CD49F+, and lin- (including mature lines CD2, CD3, CD4, CD7, CD8, CD10, CD11B, CD19, CD20, CD56, CD235A) (negative for ker). In mice, bone marrow LT-HSCs are CD34-, SCA-1+, C-kit+, CD1 35-, Slamf1 / CD150+, CD48-, and lin-(Ter119, CD11b, Gr1, CD3, CD4, CD8, B220, (Negative for mature lineage markers including IL7ra), while ST-HSC was CD34+, SCA-1+, C-kit+, CD135-, Slamf1 / CD150+, and lin-(Ter119, CD11b, Gr1, CD3, CD4, CD8, B2 20. Negative for mature strain markers including IL7ra. Furthermore, ST-HSC exhibits homeostatic symptoms. In this case, it is less inactive and more proliferative than LT-HSC. However, LT-HSC is larger They possess great self-replicating ability (i.e., they survive throughout the maturation stage and a series of recipes) ST-HSCs, on the other hand, have limited self-replication (they can be continuously transplanted through the network), while ST-HSCs have limited self-replication (they can be transplanted through the network). However, they only survive for a limited time and do not have the ability to be transplanted repeatedly. The discrepancies can also be used in the method described herein. ST-HSC is highly proliferative. Therefore, it is particularly useful because it can produce differentiated offspring more quickly.

[0159] As used herein, the term “hematopoietic stem cell functional capacity” includes hematopoietic stem cells including the following: This refers to the functional characteristics of granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils) ), red blood cells (e.g., reticulocytes, red blood cells), platelets (e.g., megakaryocytes, platelet-producing meganuclei) Spheres (platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglial cells, osteoclasts , and lymphocytes (including, but not limited to, NK cells, B cells, and T cells, for example). 1) The ability to differentiate into multiple different bloodlines), 2) Self-renewal (hematopoietic stem cells are identical to the parent cell) This refers to the ability to produce daughter cells with such abilities, and furthermore, this ability does not run out. (which can appear repeatedly throughout the life of an individual), as well as 3) hematopoietic stem cells or The offspring are reintroduced into the transplant recipient, where they home to the hematopoietic stem cell niche. The ability to restore productive and sustainable hematopoiesis.

[0160] As used herein, the term “human antibody” means substantially all parts of a protein. For example, all CDRs, framework areas, C L , C H Domain (for example, C H 1. C H 2, C H 3) Hinge, and V L and V H The domain is substantially non-immunogenic in humans, and This refers to antibodies that have only slight sequence changes or mutations. Human antibodies are antibodies that, in human cells, For example, by recombinant expression, or by functionally rearranged human immunoglobulins (heavy Non-human animals or prokaryotes capable of expressing genes (such as chains and / or light chains) It can be produced by living cells or eukaryotic cells. In combination, it may contain linker peptides not found in natural human antibodies. For example, Fv is Two to approximately eight glycine or other amino acids connect the variable region of the heavy chain to the variable region of the light chain. It may contain linker peptides such as residues. Such linker peptides are of human origin. It is considered that human antibodies are obtained using an antibody library derived from human immunoglobulin sequences. By various methods known in the art, including phage display methods Human antibodies can be produced. Human antibodies can express functional endogenous immunoglobulins. However, there are transgenic mice that can express human immunoglobulin genes. It can also be used to produce (e.g., PCT publication numbers WO 1998 / 24893; WO 1992 / 01047; W O 1996 / 34096; WO 1996 / 33735; US 5,413,923; 5,625,126; 5,633,425; 5,569,82 Details 5;5,661,016;5,545,806;5,814,318;5,885,793;5,916,771; and 5,939,598 (See book).

[0161] As used herein, the term “humanized” antibody is derived from non-human immunoglobulins. This refers to non-human antibodies containing the smallest sequence. Generally, humanized antibodies contain at least one, typically two, sequences. It includes substantially all of one variable domain, where all or substantially all of the CDR area, Corresponds to non-human immunoglobulins. The entire or substantially entire FW region is also human. It may be an immunoglobulin sequence. Humanized antibodies are immunoglobulin constant region (Fc) At least some of these may include, typically, those of human immunoglobulin consensus sequences. Methods for humanizing the body are known in this field, for example, Riechmann et al., Na ture 332:323-7, 1988; US 5,530,101; 5,585,089; 5,693,761; 5,693,762; o This is explained in specifications 6,180,370, etc.

[0162] As used herein, a patient who “needs” a hematopoietic stem cell transplant will have one or more blood cells. Patients exhibiting defects or deficiencies in the fluid cell type, as well as stem cell disorders, autoimmune diseases, cancer, and This includes patients with other medical conditions described herein. Hematopoietic stem cells are generally 1) pluripotent. It exhibits characteristics, and therefore, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), red blood Spheres (e.g., reticulocytes, red blood cells), platelets (e.g., megakaryocytes, platelet-producing megakaryocytes, blood cells) Plaques), monocytes (e.g., monocytes, macrophages), dendritic cells, microglial cells, osteoclasts, and Multiple lymphocytes, including but not limited to lymphocytes (e.g., NK cells, B cells, and T cells). 1) It can differentiate into different bloodlines, and 2) it exhibits self-renewal, and therefore is equivalent to the parent cell. It is possible to generate daughter cells with the ability, and 3) reintroduce them into the transplant recipient. They demonstrate the ability to home in the hematopoietic stem cell niche, and produce productive and sustainable results. To restore hematopoiesis. Therefore, to reconstruct defective or deficient cell populations in vivo. Therefore, hematopoietic stem cells are administered to patients who have defects or deficiencies in one or more hematopoietic cell types. It is possible. For example, a patient may have cancer, and the deficiency may be selective. Alternatively, by administering chemotherapy agents or other drugs that nonspecifically deplete the cancer cell population. It may occur. In addition or alternatively, the patient may have sickle cell anemia, thalassemia, or Fanconiosis. Anemia, aplastic anemia, and abnormal hemoglobin such as Wiscott-Aldrich syndrome The subject may have a disease (for example, non-malignant abnormal hemoglobinopathy). Nosine deaminase severe combined immunodeficiency (ADA SCID), HIV / AIDS, metachromatic leukodystrophy He suffered from Diamond-Blackfan anemia and Schwarman-Diamond syndrome. The subject may have a hereditary blood disorder (e.g., sickle cell anemia). They may have an autoimmune disorder or be affected by one. Alternatively, the subject may have a malignant tumor such as neuroblastoma or hematological cancer. or may be affected by them. For example, the subject may have leukemia, lymphoma, Or they may have myeloma. In some embodiments, the subject has acute myeloid leukemia. Blood diseases, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, They have diffuse large B-cell lymphoma or non-Hodgkin lymphoma. In the embodiment, the subject suffers from myelodysplastic syndrome. In some embodiments, the subject The body may have scleroderma, multiple sclerosis, ulcerative colitis, Crohn's disease, type 1 diabetes, or as specified herein. They suffer from an autoimmune disease, such as another autoimmune condition described in [reference]. In some embodiments, The subject requires chimeric antigen receptor T cell (CART) therapy. Several embodiments Therefore, the subject either suffers from a metabolic storage disorder or is affected by one. The subject has the following conditions, or may otherwise be affected by them: Lycogen storage disorder, mucopolysaccharidosis, Gaucher disease, Haller's disease, sphingolipidosis, dysphagia Metabolic disorders selected from the group consisting of chromatic leukodystrophy, or, not limited to, Other treatments and therapies that may benefit from those disclosed herein include, listed below. Disease or disorder: Severe combined immunodeficiency, Wiscott-Aldrich syndrome, hyperimmunoglobulinia Brin M (IgM) syndrome, Chediak-Higashi syndrome, hereditary lymphohistiocytosis, marble Bone disease, osteogenesis imperfecta, stolic disease, thalassemia major, sickle cell disease, systemic sclerosis, systemic Juvenile lupus erythematosus, multiple sclerosis, juvenile rheumatoid arthritis, and "Bone Marrow Transpl "Annotation for Non-Malignant Disease," ASH Education Book, 1:319-338 (2000) A disease or disorder which is treated by hematopoietic stem cell transplantation therapy, and the disclosure thereof is that it can be cured by the implementation of hematopoietic stem cell transplantation therapy. The entire text relating to treatable medical conditions is incorporated herein by reference. In addition to or alternatively, patients who “need” hematopoietic stem cell transplantation have one of the aforementioned medical conditions. Whether or not you have the disease, you still have megakaryocytes, platelets, red Blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglial cells, granulocytes, monocytes, osteoclasts Cells, antigen-presenting cells, macrophages, dendritic cells, natural killer cells, T lymphocytes, and reduced levels of one or more endogenous cell types within the hematopoietic lineage, such as B lymphocytes (for example) A patient may exhibit (otherwise, compared to the levels of a healthy subject). Those skilled in the art will see that In healthy subjects, the levels of one or more of the aforementioned cell types or other blood cell types decreased. To determine whether this is the case, for example, among other procedures known in the relevant technical field, flow size This can be easily determined using tometry and fluorescence-activated cell sorting (FACS) methods. It is possible.

[0163] As used herein, the term “monoclonal antibody” means the way in which it is produced. It does not originate from any eukaryote, prokaryote, or single clone including any phage clone. It refers to antibodies that produce [the substance].

[0164] As used herein, the term “recipient” means a graft containing a population of hematopoietic stem cells. This refers to patients who receive grafts such as autologous cells. The transplanted cells administered to the recipient are, for example, autologous cells. It may be a cell, a syngeneic cell, or an allogeneic cell.

[0165] As used herein, the term "sample" means a specimen taken from a subject (e.g., blood). , blood components (e.g., serum or plasma), urine, saliva, amniotic fluid, cerebrospinal fluid, tissues (e.g., fetus This refers to the disc or dermis, pancreatic juice, chorionic villi sample, and cells.

[0166] As used herein, the term "scFV" refers to the variable domain of heavy and light chains derived from an antibody. This refers to a single-stranded Fv antibody in which phosphorus molecules are linked together to form a single chain. scFV fragments are phosphorus molecules. Variable region of antibody light chain (V) separated by Kerr L )(For example, CDR-L1, CDR-L2, and / or CDR-L3) and the variable region of the antibody heavy chain (V H )(For example, CDR-H1, CDR-H2, and / or It contains a single polypeptide chain including CDR-H3. V of the scFV fragment L and V H region The linker connecting them may be a peptide linker composed of protein-generated amino acids. The alternative linker is designed to increase the resistance of scFV fragments to proteolysis. For example, a linker containing D-amino acids, to increase the solubility of scFV fragments ( For example, a polyethylene glycol-containing linker or repeated glycine and serine residues Hydrophilic linkers such as polypeptides that are contained in the molecule improve the biophysical stability of the molecule. For example, containing cysteine ​​residues that form intramolecular or intermolecular disulfide bonds. Linkers that do this, or to weaken the immunogenicity of scFV fragments (e.g., glycosylation A linker containing a linking site can be used. The variable region of the scFV molecule described herein The region can be modified so that its amino acid sequence differs from that of the antibody molecule from which it originates. This will also be understood by those skilled in the art. For example, conservative substitutions or changes in amino acid residues Substitute nucleotide or amino acid substitutions (e.g., CDR and / or frameworks) (At the residue) it retains the ability of scFV to bind to the antigen recognized by the corresponding antibody. It can be implemented to enhance it.

[0167] As used herein, the terms “subject” and “patient” are defined as follows: This refers to a living organism, such as a human, receiving treatment for a specific disease or condition. For example, a human patient. Patients such as those with conditions requiring treatment before hematopoietic stem cell transplantation are treated to promote the engraftment of exogenous hematopoietic stem cells. You may receive medical treatment.

[0168] As used herein, the phrase “substantially removed from the blood” means therapeutic agents (anti-CD45, Anti-CD135, anti-CD34, anti-CD90, or anti-CD110 antibody, its antigen-binding fragment, or rigor At some point after administration to patients (such as endothelial pneumonia), the treatment is found in a blood sample isolated from the patient. When the concentration of the drug is such that the therapeutic agent cannot be detected by conventional means. (For example, if the drug exceeds the noise threshold of the device or assay used to detect the drug) This refers to cases where detection is not performed. Various techniques known in the art, such as ELISA-based detection assays, have been found to be effective against Antibodies can be used to detect bodies, antibody fragments, and protein ligands. It can be used to detect antibody fragments and protein ligands. The assay, among those known in the art, is an immunoprecipitation technique and an immunoblock assay. Includes assay.

[0169] As used herein, the term "stem cell disorder" refers to a condition in which the target tissue of the subject is conditioned. By performing and / or excising the endogenous stem cell population in the target tissue (For example, by removing endogenous hematopoietic stem cells or progenitor cell populations from the subject's bone marrow tissue) (by doing so) and / or to engraft or transfer stem cells into the target tissue of the subject. This broad term refers to any disease, disorder, or condition that can be treated or cured by planting. For example, it has been shown that type 1 diabetes can be cured by hematopoietic stem cell transplantation, and It is possible to benefit from conditioning according to the compositions and methods described in the details. It has properties. Further disorders that can be treated using the compositions and methods described herein. The harmful effects include sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiscot's anemia. Aldrich syndrome, ADA SCID, HIV / AIDS, metachromatic leukodystrophy, diamond- Blackfan anemia and Schwarman-Diamond syndrome are among the conditions that can be treated with these. Not limited to the patient conditioning and / or hematopoietic stem cell transfer described herein. Further diseases that can be treated using the implantation method include hereditary blood disorders (e.g., sickle (Asthmatic anemia), as well as scleroderma, multiple sclerosis, ulcerative colitis, and Crohn's disease, etc. Autoimmune disorders are one example. Conditioning and / or transplantation methods described herein. Further diseases that can be treated using this method include malignant tumors such as neuroblastoma, or white blood cells. This includes blood cancers such as hematological cancers, lymphomas, and myeloma. For example, cancer is acute myeloid cancer. Leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, It may be diffuse large B-cell lymphoma or non-Hodgkin lymphoma. Further diseases treatable using the conditioning and / or transplantation methods described above include: Myelodysplastic syndromes are included. In some embodiments, the subjects suffer from metabolic storage disorders. Either they have it, or they are affected by it. For example, the subject has the following: Alternatively, you may be affected by these conditions: glycogen storage, mucopolysaccharide From disease, Gaucher disease, Haller disease, sphingolipidosis, and metachromatic leukodystrophy Metabolic disorders selected from the following groups, or, but not limited to the following, are disclosed. Another disease or disorder that may benefit from treatment and therapy: Severe combined immunodeficiency, Wiscott-Aldrich syndrome, hyperimmune globulin M (IgM) syndrome, Chediac Higashi syndrome, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, saccharomycetes, thalassemia Amygdala, sickle cell disease, systemic sclerosis, systemic lupus erythematosus, multiple sclerosis, young Rheumatoid arthritis, and "Bone Marrow Transplantation for Non-Malignant Disease," A disease or disorder described in ASH Education Book, 1:319-338 (2000), and The disclosure relates to medical conditions that can be treated by hematopoietic stem cell transplantation therapy, and The entire text is incorporated herein by reference.

[0170] As used herein, the terms “to treat” or “to treat” mean that the purpose is desired It is to prevent or slow down (reduce) any physiological changes or disorders, or This refers to therapeutic interventions that promote beneficial phenotypes in patients receiving treatment. Or, desirable clinical outcomes include the antibody conditioning therapies described herein and their One example is the promotion of engraftment of exogenous hematopoietic cells in patients following hematopoietic stem cell transplantation therapy. However, these are not the only benefits. Further beneficial results may include patients requiring hematopoietic stem cell transplantation. Conditioning therapy and subsequent placement of exogenous hematopoietic stem cell grafts in patients Examples include an increase in the number or relative concentration of hematopoietic stem cells after administration. The treatments described herein Beneficial results include the effects of conditioning therapy and subsequent hematopoietic stem cell transplantation. Megakaryocytes, platelets, platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, Microglia, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natur The number of cells of one or more hematopoietic cells, such as RAL killer cells, T lymphocytes, or B lymphocytes. Or it may include an increase in relative concentration. Further beneficial results include cancer cells (e.g., CD45 +, CD135+, CD34+, CD90+, or CD110+ leukemia cells) or autoimmune cells (for example, CD45+, CD135+, CD34+, CD90+, or expressing T cell receptors that cross-react with autoantigens. CD110+ T cells and other CD45+, CD135+, CD34+, CD90+, or CD110 autoimmune lymphocytes. This may also include a decrease in the amount of disease-causing cell populations, such as certain groups of cells.

[0171] As used herein, the terms “mutant” and “derivative” are interchangeable. The compounds, peptides, proteins, or other substances described herein, whether natural, synthetic, and This refers to semi-synthetic analogs. Compounds, peptides, proteins, or other compounds described herein. Variants or derivatives of a substance may retain or improve the biological activity of the original material. be.

[0172] As used herein, the term “vector” includes plasmids, DNA vectors, and positive This includes nucleic acid vectors such as mids, RNA vectors, viruses, or other suitable replicons. The expression vectors described herein include polynucleotide sequences, as well as, for example, proteins. The expression of nucleotides and / or the integration of these polynucleotide sequences into the genome of mammalian cells. The antibody and antibody fragment of the present invention may include additional sequence elements used for incorporation. Certain vectors that can be used for expression include promoters that direct gene transcription and Plasmids containing regulatory sequences such as enhancer regions. Antibodies and antibody fragments. Other useful vectors for gene expression enhance the translation rate of these genes, or Alternatively, polynucleotide combinations that improve the stability or nuclear export of mRNA resulting from gene transcription. It contains sequences. These sequence elements are, for example, efficient genes carried by expression vectors. It may include 5' and 3' untranslated regions and a polyadenylation signaling site for directing transcription. The expression vectors described herein are for selecting cells containing such vectors. It may contain polynucleotides that encode markers. Examples of suitable markers include amphi Antibiotics such as syrin, chloramphenicol, kanamycin, and knowulseotricin Examples include genes that code for tolerance to quality.

[0173] As used herein, the term "alkyl" means, for example, a chain containing 1 to 20 carbon atoms. This refers to linear or branched alkyl groups having n- Propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl Examples include isopentyl, tert-pentyl, hexyl, and isohexyl.

[0174] As used herein, the term "alkylene" refers to a linear or branched divalent alkyl group. Refers to a group. The divalent position can be on the same or different atoms within the alkyl chain. Examples of ethylene include methylene, ethylene, propylene, and isopropylene.

[0175] As used herein, the term "heteroalkyl" means, for example, a chain containing 1 to 20 carbon atoms. It has elementary atoms, and further contains one or more heteroatoms in the chain (for example, oxygen, nitrogen, and This refers to a linear or branched alkyl group containing sulfur.

[0176] As used herein, the term “heteroalkylene” means a divalent heteroalkylene, either linear or branched. This refers to a teloalkyl group. The divalent position is on the same or different atoms within the heteroalkyl chain. Yes, it's possible. The divalent position can be occupied by one or more heteroatoms.

[0177] As used herein, the term "alkenyl" means, for example, a chain containing 2 to 20 carbon atoms. This refers to a linear or branched alkenyl group that has offsprings. Examples of alkenyl groups include vinyl and pro. Examples include penyl, isopropenyl, butenyl, tert-butyrenyl, and hexenyl.

[0178] As used herein, the term "alkenylene" refers to a straight-chain or branched-chain divalent alkenylene. This refers to the nyl group. The divalent position can be on the same or different atoms within the alkenyl chain. Examples of luchenylenes include etenylene, propenylene, isopropenylene, and butenylene. It can be listed.

[0179] As used herein, the term “heteroalkenyl” means, for example, 2 to 20 in a chain It has carbon atoms, and further contains one or more heteroatoms in the chain (for example, oxygen, nitrogen, and more This refers to a linear or branched alkenyl group containing sulfur.

[0180] As used herein, the term "heteroalkenylene" refers to a linear or branched divalent alkenylene. This refers to a heteroalkenyl group. The divalent position indicates the same or different origin within the heteroalkenyl chain. It can be on a child. The divalent position can be one or more heteroatoms.

[0181] As used herein, the term "alkynyl" means, for example, a chain containing 2 to 20 carbon atoms. This refers to a linear or branched alkynyl group that has children. An example of an alkynyl group is propargyl. Examples include butynyl, pentynyl, and hexynyl.

[0182] As used herein, the term "alkynylene" refers to a straight-chain or branched-chain divalent alkyl This refers to the nyl group. The divalent position can be on the same or different atoms within the alkynyl chain.

[0183] As used herein, the term “heteroalkynyl” means, for example, 2 to 20 in a chain It has carbon atoms, and further contains one or more heteroatoms in the chain (for example, oxygen, nitrogen, and more This refers to a linear or branched alkynyl group containing sulfur.

[0184] As used herein, the term "heteroalkylylene" refers to a divalent linear or branched chain. This refers to a heteroalkynyl group. The divalent position indicates the same or different origin within the heteroalkynyl chain. It can be on a child. The divalent position can be one or more heteroatoms.

[0185] As used herein, the term "cycloalkyl" means saturated, e.g., 3-12 This refers to a monocyclic, condensed, bridged, or spiropolycyclic ring structure having multiple carbon ring atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclopropyl. Examples include lohexyl, cycloheptyl, cyclooctyl, and bicyclo[3.1.0]hexane. It can be done.

[0186] As used herein, the term "cycloalkylene" means a divalent cycloalkyl group To point to. The divalent position can be on the same or different atoms within the ring structure. Cycloalkylene Examples include cyclopropylene, cyclobutylene, cyclopentylene, and cyclohexylene. Some examples include:

[0187] As used herein, the term "heterocycloalkyl" means saturated, for example, From carbon atoms and heteroatoms selected from, for example, nitrogen, oxygen, and sulfur in particular. Monocyclic, condensed, or bridging rings, each having 3 to 12 selected ring atoms per ring structure. This refers to a spiropolycyclic ring structure. The ring structure is, for example, formed on the ring members of carbon, nitrogen, or sulfur. It may contain one or more oxo groups.

[0188] As used herein, the term "heterocycloalkylene" means a divalent heterocyclo Refers to an alkyl group. The divalent position can be on the same or different atoms within the ring structure. The position can be occupied by one or more heteroatoms.

[0189] As used herein, the term “aryl” means, for example, containing 6 to 19 carbon atoms. This refers to monocyclic or polycyclic aromatic ring systems. Aryl groups include phenyl, fluorenyl, and na. Examples include phthyl, but the list is not limited to these.

[0190] As used herein, the term "arylene" refers to a divalent aryl group. The positions can be on the same or different atoms.

[0191] As used herein, the term "heteroaryl" means monocyclic heteroaromatic, or This refers to bicyclic or tricyclic fused ring heteroaromatic groups. Examples of heteroaryl groups include the following: Possible compounds: pyridyl, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, iso Oxazolyl, thiazolyl, isothiazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4- Triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadia Zolyl, 1,3,4-oxadiazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl, benzof Lyl, [2,3-dihydro]benzofuryl, isobenzofuryl, benzothienyl, benzotri Zolyl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzyl Midazolyl, imidazo[1,2-a]pyridyl, benzothiazolyl, benzoxazolyl, quinol Ridinyl, quinazolinyl, phthalazinyl, quinoxalinyl, sinnolinyl, naphthilidinyl Lu, pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4,3-b]pyridyl, quinori Lu, isoquinolyl, tetrazolyl, 5,6,7,8-tetrahydroquinolyl, 5,6,7,8-tetrahydro Roisoquinolyl, Purinyl, Pteridinyl, Carbazolyl, Xanthenyl, Benzoquinolyl Ru, etc.

[0192] As used herein, the term "heteroarylene" refers to a divalent heteroaryl group. The divalent position can be on the same or different atoms. The divalent position can be on one or more hetagonal atoms. It could be a 'ro' atom.

[0193] Unless otherwise specifically limited by the definition of individual substituents, the aforementioned chemical sites, for example, "al "Kil", "alkylene", "heteroalkyl", "heteroalkylene", "alkenyl" "Alkenylene", "Heteroalkenyl", "Heteroalkenylene", "Alkinyl" "Alkynylene", "Heteroalkynyl", "Heteroalkynylene", "Cycloalkyl "Cycloalkylene", "Heterocycloalkyl", "Heterocycloalkylene" "aryl", "arirene", "heteroaryl", and "heteroarirene" groups Examples include alkyl, alkenyl, alkynyl, cycloalkyl, and heterocycloalkyl. Alkylaryl, alkylheteroaryl, alkylcycloalkyl, alkylhe Terocycloalkyl, amino, ammonium, acyl, acyloxy, acylamino, a Minocarbonyl, alkoxycarbonyl, ureido, carbamate, aryl, hetero Lille, sulfinyl, sulfonyl, alkoxy, sulfanyl, halogen, carboxy Selected from the group consisting of trihalomethyl, cyano, hydroxy, mercapto, nitro, etc. It may be optionally substituted with 1 to 5 substituents. Substitutions include ring closure of adjacent functional substituents. , adjacent substituents undergo ring closure, for example, forming lactams, lactones, and cyclic compounds through ring closure. Anhydrous, acetal, hemiacetal, thioacetal, aminal, and heminamine This may include situations where a ring is formed and, for example, a protecting group is supplied.

[0194] (Antibodies that recognize hematopoietic cell antigens) This invention is partly based on the following discovery: CD45, CD135, CD34, CD90, or CD110 Antibodies that can bind, their antigen-binding fragments, and ligands as therapeutic agents (i) cancer and autologous cancer characterized by cells expressing one or more of these antigens. (ii) In patients who require transplantation therapy, the immune disease can be directly treated. It can promote the engraftment of transplanted hematopoietic stem cells. These therapeutic activities are, for example, anti- The body, its antigen-binding fragments, and / or ligands, cancer cells, autoimmune cells, Alternatively, binding to one or more of the aforementioned antigens expressed on the surface of cells such as hematopoietic stem cells. This can be caused by the subsequent induction of cell death. Depletion of endogenous hematopoietic stem cells is It provides a niche where transplanted hematopoietic stem cells can home, and subsequently, productive Hematopoiesis can be established in this way. In this manner, transplanted hematopoietic stem cells are used in this specification. The stem cells can successfully engraft in human patients and other patients suffering from the stem cell disorders described below. In this section, each of these antigens, as well as antibodies that can bind to these targets, This section describes the antigen-binding fragment of biology.

[0195] (anti-CD45 antibody) Includes those that can bind to isoform CD45RO, and those that can bind to human CD45. Possible antibodies and antigen-binding fragments (mRNA NCBI reference sequence: NM_080921.3, protein) The NCBI reference sequence (NP_563578.2) is used in conjunction with the compositions and methods disclosed herein. For example, promoting the engraftment of hematopoietic stem cell grafts in patients requiring hematopoietic stem cell transplantation therapy. Multiple isoforms of CD45 allow for the selection of 34 exons in the primary transcript. It results from splicing. Splicing of exons 4, 5, 6, and potentially 7 is It causes multiple CD45 mutations. Selective exon expression is the CD45 isoform listed in Table 1 below. Observed in the form. [Table 1]

[0196] Alternative splicing involves different isoforms of the CD45 protein (e.g., CD45 Individual exons or combinations of exons expressed in RA, CD45RAB, CD45RABC This can lead to... In contrast, CD45RO lacks expression of exons 4-6, and... It is generated from combinations of exons 1-3 and 7-34. Exon 7 is also excluded from the protein. As a result, there is evidence that exons 1-3 and 8-34 are spliced ​​together. This protein, named E3-8, has been detected at the mRNA level, but in the flow charts... It has not yet been identified by tometry.

[0197] CD45RO is currently the only known CD45 isoform expressed on hematopoietic stem cells. 45RA and CD45RABC were either not detected or excluded from the hematopoietic stem cell phenotype. Yes. Studies conducted in mice have shown that CD45RB is expressed in fetal hematopoietic stem cells, but not in adult bone marrow. There is evidence that it does not exist in hematopoietic stem cells. In particular, CD45RC is found in the Asian population. Exon 6 polymorphisms are common (CD45RC exon 6 polymorphisms are found in approximately 25% of the Japanese population). This polymorphism results in high expression of CD45RO and decreased levels of CD45RA, CD45RB, and CD45RC. Furthermore, CD45RA variants (such as CD45RAB and CD45RAC) are associated with autoimmune diseases. It exhibits polymorphism in exon 4.

[0198] The presence of CD45RO on hematopoietic stem cells and other immune cells (T and B lymphocyte subsets and Its relatively limited expression on various cells (such as bone marrow cells) indicates that CD45RO is being expressed on hematopoietic stem cell transfer. CD45RO is a particularly suitable target for conditioning therapy in patients requiring plant implantation. Because it lacks expression of only Son 4, 5, and 6, its use as an immunogen is limited to the total CD45 Ab This also enables screening of CD45RO-specific antibodies.

[0199] Anti-CD45 anti- For example, commercially available products from BIOLEGEND (registered trademark) (San Diego, California) Examples include the anti-CD45 antibody clone HI30 and its humanized variant. The ionization is performed according to procedures known in the art, using framework residues of non-human antibodies. This can be done by substituting the constant region residues with those of germline human antibodies. (For example, as described in Example 7 below). It can be used in conjunction with the methods described herein. Additional anti-CD45 antibodies include those from ABCAM (registered trademark) (Cambridge, Massachusetts). The following anti-CD45 antibodies are commercially available: ab10558, EP322Y, MEM-28, ab10559, 0.N.125, F10-89- 4, HIe-1, 2B11, YTH24.5, PD7 / 26 / 16, F10-89-4, 1B7, ab154885, B-A11, Phosphor S1007, ab170444, EP350, Y321, GA90, D3 / 9, X1 6 / 99, and LT45, as well as their humanized variants. Examples include: Used in conjunction with the patient conditioning procedures described herein. Further anti-CD45 antibodies that can do this include SIGMA-ALDRICH (registered trademark) (St. Louis, Ms. Examples include the anti-CD45 antibody HPA000440, which is commercially available from the state of London, and its humanized variant. This can be used in conjunction with the patient conditioning methods described herein. For example, the anti-CD45 antibody described in Matthews et al., Blood 78:1864-1874, 1991 The mouse monoclonal antibody BC8 is mentioned, and its disclosure is an anti-CD45 antibody, as well as its The humanized variants described herein are incorporated herein by reference. Further anti-CD45 antibodies that can be used in conjunction with this method include, for example, Glatting et al. The monoclonal antibody YAML568, described in J. Nucl. Med. 8:1335-1341, 2006, is one example. The disclosure relates to an anti-CD45 antibody and its humanized variants, as can be seen by reference. This is incorporated herein. It is used in conjunction with the patient conditioning procedures described herein. Additional anti-CD45 antibodies that can be used include, for example, Brenner et al., Ann. NY Academia. The monoclonal antibodies YTH54.12 and YTH25.4 described in . Sci. 996:80-88, 2003 The disclosures cited are referenced as relating to anti-CD45 antibodies and their humanized variants. This is incorporated herein by means of. Used in conjunction with the patient conditioning methods described herein. For additional anti-CD45 antibodies to use, see, for example, Brown et al., Immunology 64:331-33. UCHL1, 2H4, SN130, MD4.3, MBI, and MT2 are listed in 6, 1998, and their The disclosure relating to anti-CD45 antibodies and their humanized variants is hereby referred to herein. It is incorporated into. Additional anti-CD45 antibodies that can be used in conjunction with the method described herein. This includes American Type Culture Collection (ATCC) accession numbers RA3-6132, RA3-2C2, and those produced and released from TIB122, as well as, for example, Johnson et al., J. E. The monoclonal antibody C363.16A and 13 / 2 described in xp. Med. 169:1179-1184, 1989 The disclosures include, and the disclosures relate to anti-CD45 antibodies and their humanized variants. This is incorporated herein by reference. In conjunction with the patient conditioning methods described herein. Further anti-CD45 antibodies that can be used include, for example, Harvath et al., J. Immunol The monoclonal antibodies AHN-12.1, AHN-12, and AHN-12.2 are described in . 146:949-957, 1991. Examples include AHN-12.3, AHN-12.4, HLe-1, and KC56(T200), and their disclosure is anti-CD45 anti The body, and its humanized variants, are incorporated herein by reference.

[0200] Additional antimicrobial agents that can be used in conjunction with the patient conditioning methods described herein. CD45 antibodies include, for example, U.S. Patent No. 7,265,212 (for example, anti-CD45 among other clones). Antibodies 39E11, 16C9, and 1G10 are listed); 7,160,987 (e.g., monoclonal antibody 6G (Describe anti-CD45 antibodies produced and released by ATCC accession number HB-11873, such as 3.) ;and 6,099,838 (anti-CD45 antibody MT3), and ATCC accession number HB220 (also designated as MB23G2) (and describes antibodies produced and released by HB223), and U.S. Patent U.S. Patent No. 2004 / 0096901 and U.S. Patent No. 2008 / 0003224 (for example, ATCC accession number PTA-7339) Therefore, the anti-CD45 antibodies produced and released (for example, monoclonal antibodies 17.1) The following are listed, and each of their disclosures is by reference as relating to anti-CD45 antibodies. This specification is incorporated herein.

[0201] Further methods that can be used in conjunction with the patient conditioning methods described herein Anti-CD45 antibodies include ATCC accession numbers MB4B4, MB23G2, 14.8, GAP 8.3, 74-9-3, I / 24.D6, 9. Antibodies produced and released from 4, 4B2, M1 / 9.3.4.HL.2, as well as their humanization and / or affinity-maturing mutants. Affinity maturation is described, for example, in Example 6 below. As such, phage displays and other applications described herein or in the relevant technical field This can be accomplished using known in vitro display techniques.

[0202] Additional antimicrobial agents that can be used in conjunction with the patient conditioning methods described herein. CD45 antibodies are described, for example, in Morikawa et al., Int. J. Hematol. 54:495-504, 1991. The anti-CD45 antibody T29 / 33 is mentioned, and its disclosure is referenced as relating to anti-CD45 antibodies. This is incorporated herein by means of.

[0203] Each of the disclosures in the aforementioned publications is made known by reference as relating to anti-CD45 antibodies. To be incorporated into the specifications. Antibodies that may be used in conjunction with the compositions and methods described herein. The antigen-binding fragments include the antibodies and their antigen-binding fragments, as well as the above Humanized variants of the non-human antibodies and antigen-binding fragments described below, and, for example, competitive CD Antibodies that bind to the same epitopes as those described above, as evaluated by the 45-binding assay. Alternatively, antigen-binding fragments may be used.

[0204] (anti-CD135 antibody) This invention presents an antibody capable of binding to CD135 and its antigen-binding fragment (Flt2 (Also known as Flt3) is used as a therapeutic agent to directly treat cancer and autoimmune diseases. This can promote the engraftment of transplanted hematopoietic stem cells in patients requiring transplantation therapy. This is partly based on the discovery that it is possible. Furthermore, CD13 such as human Flt3 ligand By using ligands that bind to 5, cancer and autoimmune diseases can be directly treated. Furthermore, it can promote the engraftment of transplanted hematopoietic stem cells in patients requiring transplantation therapy. It was discovered that these ligands, such as human Flt3 ligands, are, for example, antibody-dependent. Effector domains such as the Fc domain promote cell-mediated cytotoxicity (ADCC). It can be covalently bonded to it.

[0205] Hematopoietic stem cells have been shown to express CD135 because this antigen is an a insulator of Mcl-1. It has been reported that this promotes supregulation, which in turn regulates cell survival during hematopoiesis. This is because it is a receptor tyrosine kinase (for example, its disclosure is due to hematopoietic stem cells) The expression of CD135 is incorporated herein by reference in J. Immunol. 180:7 (See 358-7367, 2008). Antibodies capable of binding to this cell surface antigen, and their antigen binding. Fragments and ligands are known in the art and described herein. Using techniques such as immunization, computational modeling, and phagedis described below, In vitro selection methods such as play and cell-based display platforms It can be identified as such.

[0206] Anti-CD135 which can be used in conjunction with the patient conditioning methods disclosed herein. The antibodies may include those possessing one or more, or all, of the following CDRs: a. CDR-H1 having the amino acid sequence SYYMH (SEQ ID NO: 1); b. CDR-H2 having the amino acid sequence IINPSGGSTSYAQKFQG (SEQ ID NO: 2); c. CDR having amino acid sequence GVGAHDAFDI (SEQ ID NO: 3) or VVAAAVADY (SEQ ID NO: 4) -H3; d. Amino acid sequence RSSQSLLHSNGNNYLD (SEQ ID NO: 5) or RSSQSLLHSNGYNYLD (SEQ ID NO: 6) ) CDR-L1; e. CDR-L2 having the amino acid sequence LGSNRAS (SEQ ID NO: 7); and f. CDR- having amino acid sequence MQGTHPAIS (SEQ ID NO: 8) or MQSLQTPFT (SEQ ID NO: 9) L3.

[0207] Additional antimicrobial agents that can be used in conjunction with the patient conditioning methods disclosed herein. CD135 antibodies include those containing one or more, or all, of the following CDRs: a. CDR-H1 having the amino acid sequence SYAIS (SEQ ID NO: 10); b. CDR-H2 having the amino acid sequence GIIPIFGTANYAQKFQG (SEQ ID NO: 11); c. CDR-H3 having the amino acid sequence FALFGFREQAFDI (SEQ ID NO: 12); d. CDR-L1 having the amino acid sequence RASQSISSYLN (SEQ ID NO: 13); e. CDR-L2 having the amino acid sequence AASSLQS (SEQ ID NO: 14); and f. CDR-L3 having the amino acid sequence QQSYSTPFT (SEQ ID NO: 15).

[0208] The aforementioned antibody is described, for example, in U.S. Patent No. 8,071,099, and its disclosure is anti-CD1 35 antibodies and their antigen-binding fragments are incorporated herein by reference. Antibodies such as IMC-EB10 and IMC-NC7, disclosed in U.S. Patent No. 8,071,099, are used. The fragments and their components can be used in conjunction with the methods disclosed herein. .

[0209] In addition to the above, it may be used in conjunction with the patient conditioning methods described herein. The anti-CD135 antibodies that can be produced include those that have one or more, or all, of the following CDRs. ru: a. CDR-H1 having the amino acid sequence SYWMH (SEQ ID NO: 22); b. CDR-H2 having the amino acid sequence EIDPSSDSYKDYNQKFK (SEQ ID NO: 23); c. CDR-H3 having the amino acid sequence AITTTPFDF (SEQ ID NO: 24); d. CDR-L1 having the amino acid sequence RASQSISNNLH (SEQ ID NO: 25); e. CDR-L2 having the amino acid sequence YASQSIS (SEQ ID NO: 26); and f. CDR-L3 having the amino acid sequence QQSNTWPYT (SEQ ID NO: 27).

[0210] Additional The anti-CD135 antibodies include those containing one or more, or all, of the following CDRs: a. CDR-H1 having the amino acid sequence NYGLH (SEQ ID NO: 28); b. CDR-H2 having the amino acid sequence VIWSGGSTDYNAAFIS (SEQ ID NO: 29); c. CDR-H3 having the amino acid sequence KGGIYYANHYYAMDY (SEQ ID NO: 30); d. CDR-L1 having the amino acid sequence KSSQSLLNSGNQKNYM (SEQ ID NO: 31); e. CDR-L2 having the amino acid sequence GASTRES (SEQ ID NO: 32); and f. CDR-L3 having the amino acid sequence QNDHSYPLT (SEQ ID NO: 33).

[0211] The aforementioned antibody is described, for example, in U.S. Patent No. 9,023,996, which discloses anti-CD1 35 antibodies and their antigen-binding fragments are incorporated herein by reference. It is produced. Antibodies disclosed in U.S. Patent No. 9,023,996, such as clones 4G8 and BV10. The fragments can be used in conjunction with the methods disclosed herein.

[0212] Other anti-CD agents that can be used in conjunction with the patient conditioning methods described herein. 135 antibodies include, for example, American Type Culture Collection (ATCC) accession number ATCC It contains antibodies produced and released by HB 11,557, which are, for example, U.S. 5,6 This is described in publication number 35,388, and its disclosure relating to anti-CD135 antibodies is by reference to this publication. It will be incorporated into the specification. It will be used in conjunction with the patient conditioning methods described herein. Additional anti-CD135 antibodies that can be used include, for example, the Budapest Convention as of December 19, 1995. Based on the number DSM ACC2249, the international depositary organization DSMZ-Deutsche Sammung von Mikroorga Nismen und Zellkulturen GmbH Mascheroder Weg 1b (D-38124, Braunschweig, D It was deposited in ITS and produced and released by hybridoma cells named 4G8B4B 12. The antibodies that are released are listed, for example, in U.S. Patent No. 6,156,882. That disclosure is incorporated herein by reference as relating to an anti-CD135 antibody. Additional anti-CD135 The antibody, for example, was designated as DSM-ACC2248 under the Budapest Convention on December 19, 1995. It was deposited with the German Collection of Microorganisms and Cell Cultures Ltd., and is designated BV10A4. Examples include antibodies produced and released by hybridoma cells named H2, and For example, this is described in U.S. Patent No. 5,777,084, the disclosure relating to anti-CD135 antibodies. The patient conditions described herein are incorporated herein by reference. Additional anti-CD135 antibodies that can be used in conjunction with the aging method include, for example, U.S. 7,1 The antibodies produced and released by ATCC accession number FTA-4089, as described in No. 83,385, are included. In rare cases, such disclosures are incorporated herein by reference as relating to anti-CD135 antibodies. Additional anti-CD135 antibodies are available under U.S. Patent No. 5,548,065 (e.g., ATCC accession number CRL 10907, C). Anti-CD135 antibodies produced and released by RL 10935, CRL 10936, and CRL 11005, (including antigen-binding fragments and ligands) and Patent No. 9,109,227, and U.S. Patent This is described in Patent Publication No. 2009 / 0054358, and each of these disclosures relates to anti-CD135 antibodies. It is incorporated herein by reference as a means of doing so. Used in conjunction with the methods described herein. Other anti-CD135 antibodies for this purpose are, for example, those described in U.S. Patent Application Publication No. 2010 / 0093008. As shown, antibodies produced by the rabbit hybridoma cell line C24D9, and Humanized variants are cited, and their disclosure is by reference to this book as relating to anti-CD135 antibodies. It will be included in the specifications.

[0213] In addition to the above, antibodies that can bind to CD135 and their antigen-binding fragments Examples include those described in International Publication No. 1995 / 007348, which discloses anti-CD135 anti- This is incorporated herein by reference as relating to the body. For example, ATCC accession number HB 1 Humanized variant of rat antibody 19A produced and released by 1442, ATCC accession number HB 1144 Humanized variant of antibody 23H produced and released by 3, produced by ATCC accession number 11444 and the humanized variant of antibody 2A13, produced and released by ATCC accession number HB 11445. Humanized variants of the mouse antibody 6J11 produced and rats described in International Publication No. 1995 / 007348 Humanized variants of antibody 7IE are used in conjunction with the methods described herein for hematopoietic stem cell transplantation therapy. Patients can be conditioned beforehand.

[0214] In addition to antibodies and antigen-binding fragments, CD135 ligands such as human Flt3 ligand are also included. To treat a patient, for example, cancer or an autoimmune disorder, in accordance with the method described herein. It is administered to condition the patient, either for this purpose or before hematopoietic stem cell transplantation. This is possible. For example, CD135 ligands such as human Flt3 ligand can be used (see below for example). Cytotoxins or Fc-domei (according to the method or methods known in the art) It can be conjugated to other effector molecules such as n. CD135 ligands for use in conjunction with the method include, for example, human Flt3 ligand-IgG1 Fc conjugate. Sugate, human Flt3 ligand-IgG2 Fc conjugate, human Flt3 ligand-IgG3 Fc conjugate Conjugates, and human Flt3 ligand-IgG4 Fc conjugates, e.g., ADIPOGEN (Registered (Trademark) (San Diego, California), manufactured under product number AG-40B-0119 These are some examples.

[0215] Each of the disclosures in the aforementioned publications relating to anti-CD135 antibodies is referred to herein by reference. Antibodies and antigens that may be used in conjunction with the compositions and methods described herein. The binding fragments include the antibody and its antigen-binding fragment, as well as the non- Humanized variants of human antibodies and antigen-binding fragments, and, for example, competitive CD135 binding As evaluated by the assay, antibodies that bind to the same epitopes as those mentioned above, Examples include antigen-binding fragments.

[0216] (anti-CD34 antibody) Human CD34 (mRNA NCBI reference sequence: NM_001025109.1, protein NCBI reference sequence: NP_001020) Antibodies and antigen-binding fragments that can bind to 280.1) are described herein. It can be used in conjunction with the composition and method. In a significant proportion of the population, the CD34 coding The understanding of polymorphisms affecting the cellular region or extracellular domain is not yet established. There are two isoforms of CD34 with different cytoplasmic tail lengths (long and short). In recent years, long isoforms have been used to express CD34, which can be used as an immunogen. A stable cell line was generated (see, for example, Adv. Pharm. Bull. 5:69-75, 2015). CD3 4. For example, long isoforms of CD34 can bind to CD34, for example, cancer and for identifying antibodies and their antigen-binding fragments suitable for the treatment of autoimmune diseases. It can be used as an immunogen, as well as a pre-treatment conditioner for hematopoietic stem cell transplantation. It can also be used as a lubricant.

[0217] CD34 antibodies that can be used in conjunction with the methods described herein include, for example, those used in the United States Production from ATCC accession numbers AC133.1 and HB 12346, as described in No. 5,843,633. This includes, but is not limited to, antibodies that are released.

[0218] The disclosures in the aforementioned publications relating to anti-CD34 antibodies are incorporated herein by reference. Antibodies and antigen-binding fluids that may be used in conjunction with the compositions and methods described herein. The compound contains the above-mentioned antibody and its antigen-binding fragment, as well as the above-mentioned non-human antibody. and humanized variants of antigen-binding fragments, and, for example, competitive CD34 binding assays. As evaluated by, antibodies or antigen binding that bind to the same epitopes as those mentioned above. Fragments are one example.

[0219] (anti-CD90 antibody) An antibody capable of binding to human CD90 and its antigen-binding fragment are described herein. It can be used in conjunction with the described compositions and methods. Hematopoietic stem cells are derived from human umbilical cord blood Li Since the n-CD34+CD38-CD90+CD45RA- fraction is enriched and reported to contain hematopoietic stem cells, CD It has been shown to express 90 (e.g., Majeti et al., Cell Stem Cell 1:635-645, See 2007, the disclosure relating to CD90 expression by hematopoietic stem cells, by reference. (Incorporated herein) Antibodies and antigens capable of binding to this cell surface antigen. The composite fragment is used for immunization and phage display and cell-based development. Known in the art, such as in vitro selection technologies including spray platforms. and can be produced using the techniques described herein.

[0220] Anti-CD90 antibodies that can be used in conjunction with the methods described herein include, but are not limited to, , EPR3132, EPR3133, AF-9, 5E10, F15-42-1, 7E1B11, ab189367, aTHy-1A1, ab106934, and ab110477 are examples, and these are registered trademarks of ABCAM (Cambridge, Massachusetts) These are commercially available products from the state of [unclear], as well as their humanized variants.

[0221] Antibodies and antigen-binding fragments that may be used in conjunction with the compositions and methods described herein. The reagent includes the above-mentioned antibody and its antigen-binding fragment, as well as the above-mentioned non-human antibody and Humanized variants of antigen-binding fragments, and, for example, competitive CD90 binding assays As evaluated, antibodies or antigen-binding fungibles that bind to the same epitope as those mentioned above. Gument is one example.

[0222] (anti-CD110 antibody) Antibodies capable of binding to human CD110 and their antigen-binding fragments (c-mpl and Platelet receptors (also known as TPOs) can be used in conjunction with the compositions and methods described herein. It can be used. Hematopoietic stem cells have been shown to express CD110, and the expression of CD110 is It correlates with the functional potential of hematopoietic stem cells, and the differentiation of hematopoietic stem cells into the hematopoietic lineage. (See, for example, Ninos et al., J. Transl. Med. 4:9, 2006, and its disclosure) This is incorporated herein by reference as relating to the expression of CD110 by hematopoietic stem cells. (This means that antibodies and antigen-binding fragments that can bind to this cell surface antigen are immunosuppressive.) Epidemic and the following phage display and cell-based display platforms Techniques known in the art, including in vitro selection techniques, and those described herein. It can be raised using this technology.

[0223] The cancer treatment method, autoimmune disease treatment method, and patient conditioning described herein Examples of anti-CD110 antibodies that can be used in conjunction with the methods described herein, such as the quenching method, include: For example, De Gobbi et al., Epigenetics Chromatin 4:9, 2011;Erickson-Miller et al., BMC Cancer 12:405, 2012; and Petit Cocault et al., Exp. Hematol. 44:297-302, 2 Examples include clones BAH-1, 1.78.1, and 1.6.1 described in 016, each of which disclosures are as follows: This is incorporated herein by reference as relating to an anti-CD110 antibody.

[0224] Each of the disclosures in the aforementioned publications relating to anti-CD110 antibodies is referred to herein by reference. Antibodies and antigens that may be used in conjunction with the compositions and methods described herein. The binding fragments include the antibody and its antigen-binding fragment, as well as the non- Humanized variants of human antibodies and antigen-binding fragments, and, for example, competitive CD110 binding As evaluated by the assay, antibodies that bind to the same epitopes as those mentioned above, Examples include antigen-binding fragments.

[0225] (Additional antibodies and their antigen-binding fragments) Antibodies and ligands for use in conjunction with the methods described herein shall have an Fc domain. Variants of the above antibodies, such as antibody fragments containing or lacking the antibodies described herein, and the information provided herein. One or more or all of the CDRs or their equivalents of antibodies, antibody fragments, or ligands. Humanized variants of non-human antibodies and antibody-like protein scaffolds described herein, including the region ( for example, 10 Examples include the Fn3 domain. The aforementioned example antigen-binding fragment of the antibody is In particular, the bivariable immunoglobulin domain, single-stranded Fv molecule (scFv), and diabody Tria bodies, nano bodies, antibody-like protein scaffolds, Fv fragments, Fab fragments Examples include F(ab')2 molecules and tandem di-scFv.

[0226] (Methods of combination therapy) Using the methods described herein, a physician in the field may diagnose cancer or autoimmune disease. It can treat the patient, or before hematopoietic stem cell transplantation therapy, one of the methods described herein By administering the above antibodies, their antigen-binding fragments, or ligands to the patient... Conditioning can be performed. For example, an internist can recognize CD45 (e.g., CD45RO). Antibodies that recognize and bind, their antigen-binding fragments, or ligands, either alone or An antibody that recognizes and binds to CD135, its antigen-binding fragment, or ligand, or CD34. Antibodies that recognize and bind to CD90, their antigen-binding fragments, or ligands, which recognize CD90 The binding antibody, its antigen-binding fragment, or ligand, and / or CD110 Administered in combination with an antibody that recognizes and binds to the antigen, its antigen-binding fragment, or a ligand. By doing so, cancer or autoimmune diseases can be treated, or hematopoietic stem cell transplantation can be performed. The patient may be conditioned for this purpose. There may be two or more medication plans for combination therapy. This may include sequential administration of the above-mentioned antibody, its antigen-binding fragment, or ligand, for example, The patient is administered the first antibody, its antigen-binding fragment, or ligand at the specified time. Then, the second antibody, its antigen-binding fragment, or ligand is administered to the patient at a later time. For example, administer the drug in a way that optimizes the pharmacokinetic profile of the combined therapeutic dose. Two or more antibodies that bind to one or more hematopoietic stem cell antigens described in the specification, and their antigenic binding Synthetic fragments or ligands can be administered simultaneously to patients (e.g., human patients). They may be mixed together in a single pharmaceutical composition or administered at different times. (For example, within 1 to 24 hours from each other, for example, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours from each other, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours Between 17, 18, 19, 20, 21, 22, 23, 24 hours, or longer. (Within the above time frame).

[0227] Antibodies, their antigen-binding fragments, and ligands are used to perform multiple hematopoiesis using a single antibody. It can be administered to patients to recognize and bind to stem cell antigens. This is, for example, This can be achieved using bispecific and multispecific antibodies. For example, CD45 ( For example, an antigen-binding fragment that selectively binds to CD45RO and another that selectively binds to CD135. A bispecific antibody containing an antigen-binding fragment is used in patients requiring hematopoietic stem cell transplantation therapy. It can be administered to patients, but the purpose is to deplete CD45+CD135+ cells, and thereby This creates a vacancy in hematopoietic tissue (e.g., bone marrow) that can be replaced by exogenous hematopoietic stem cell transplantation. This is to release antigenic antibodies that selectively bind to CD45 (e.g., CD45RO) and CD34. A bispecific antibody containing a combined fragment is administered to the patient before hematopoietic stem cell transplantation, along with CD34+CD4 It can be administered to deplete 5+ cells. Specifically, CD45 (e.g., CD45RO) and CD90. A bispecific antibody containing an antigen-binding fragment that selectively binds to cancer or autoimmune diseases. For patients suffering from infectious diseases, CD45+CD90+ cells are administered before hematopoietic stem cell transplantation. It can be administered similarly to deplete them. Furthermore, CD45 (e.g., CD45RO) and CD110 A bispecific antibody containing an antigen-binding fragment that selectively binds is administered to a patient, for example, It can be administered before hematopoietic stem cell transplantation to deplete CD45+CD110+ cells.

[0228] In some embodiments, an antigen-binding fragment selectively binds to CD135 and a selective antigen-binding fragment binds to CD34. A bispecific antibody containing a selectively binding antigen-binding fragment is used, for example, in cancer, autoimmune diseases. Patients suffering from autoimmune diseases, or patients requiring hematopoietic stem cell transplantation, etc. It can be administered to patients, but the purpose is to deplete CD34+CD135+ cells, thereby This creates a vacancy in hematopoietic tissue (e.g., bone marrow) that can be filled with exogenous hematopoietic stem cell transplantation. This is to release them. Similarly, antigen-binding fragments that selectively bind to CD135 and CD90 The bispecific antibody contained in the antibody was administered to the patient to deplete CD90+CD135+ cells before hematopoietic stem cell transplantation. It can be administered to induce [a specific condition]. Antigen-binding flag that selectively binds to CD135 and CD110. A bispecific antibody containing ment is administered to the patient, for example, before hematopoietic stem cell transplantation therapy, such as CD110+CD It can be administered similarly to deplete 135+ cells.

[0229] In some embodiments, an antigen-binding fragment selectively binds to CD34 and selectively binds to CD90. A bispecific antibody containing another antigen-binding fragment that binds to it is used for cancer, autoimmune diseases, and other conditions. It can be administered to patients suffering from a disease or patients requiring hematopoietic stem cell transplantation. However, the purpose is to deplete CD34+CD90+ cells, thereby reducing hematopoietic tissue (e.g., bone marrow). This is to create a vacancy that can be filled by exogenous hematopoietic stem cell transplantation. Similarly, CD A bispecific antibody containing antigen-binding fragments that selectively bind to 34 and CD110 is administered to the patient. For example, administering it to deplete CD34+CD110+ cells before hematopoietic stem cell transplantation. It is possible.

[0230] In some embodiments, an antigen-binding fragment selectively binds to CD90 and CD110. A bispecific antibody containing a selectively binding antigen-binding fragment is used for cancer and autoimmune diseases. It can be administered to patients suffering from infectious diseases or patients requiring hematopoietic stem cell transplantation. However, the purpose is to deplete CD90+CD110+ cells, thereby reducing hematopoietic tissue (e.g., bone marrow). This is to create vacancies that can be filled with exogenous hematopoietic stem cell transplantation.

[0231] (Methods for identifying antibodies and ligands) The parts that can be combined with CD45 (e.g., CD45RO), CD135, CD34, CD90, and CD110 High-throughput antibody, antibody fragment, and ligand libraries for this product. Using methods for screening, and for treating cancer, autoimmune diseases, and as specified herein As described, patients requiring hematopoietic stem cell therapy (e.g., human patients) are conditioned This method allows for the identification of affinity-mature antibodies that are useful for [specific purpose]. Phage displays, bacterial displays, yeast displays, mammalian cell displays This includes displays such as ribosome displays, mRNA displays, and cDNA displays. Examples include in vitro display technologies known in the art. Biologically related The use of phage display to isolate ligands that bind to molecules is, for example, Fe lici et al., Biotechnol. Annual Rev. 1:149-183, 1995;Katz, Annual Rev. Biophys. Biomol. Struct. 26:27-45, 1997; and Hoogenboom et al., Immunotechnology 4:1-20 Reviewed in 1998, each of these disclosures relates to in vitro display technology. This is incorporated herein by reference as a drug discovery method. Kay, Perspect. Drug Discovery De This is described in s. 2:251-268, 1995 and Kay et al., Mol. Divers. 1:139-140, 1996. To select polypeptides that bind to cell surface antigens, randomized combinatorial combinations are used. A real peptide library has been constructed, and each disclosure of these is an antigen-binding molecule. This is incorporated herein by reference as relating to the discovery of multimeric proteins and other Proteins are successfully phage-displayed as functional molecules (e.g., EP 0) 349578;EP 4527839;and EP 0589877, as well as Chiswell and McCafferty, Trends Bi See otechnol. 10:80-84 1992, each of which disclosures is for the discovery of antigen-binding molecules. This is incorporated herein by reference as relating to the use of in vitro display technology. Furthermore, functional antibody fragments such as Fab fragments and scFV fragments. It has been expressed in an in vitro display format (e.g., McCafferty et al., Nature 348:552- 554, 1990; Barbas et al., Proc. Natl. Acad. Sci. USA 88:7978-798 See 2, 1991; and Clackson et al., Nature 352:624-628, 1991, and each of these The disclosure relates to the use of in vitro display technology for the discovery of antigen-binding molecules. (and incorporated herein by reference). These technologies, among others, include hematopoietic stem cell antigens. Antibodies that bind to CD45 (e.g., CD45RO), CD135, CD34, CD90, and CD110. It can be used to identify and improve affinity, which then affects hematopoietic stem cell transfer. To deplete endogenous hematopoietic stem cells in patients requiring graft therapy (e.g., human patients) It can be used for that purpose.

[0232] In addition to in vitro display technology, computer modeling technology is used to create CD45 Antibodies that bind to (for example, CD45RO), CD135, CD34, CD90, or CD110, antibody fragments The bristles and ligands can be designed and identified in silico. For example, a computer... Using antibody modeling techniques, those skilled in the art can model antibodies, antibody fragments, and ligands. The library, in silico, CD45 (e.g., CD45RO), CD135, CD34, CD90, and Molecules that can bind to specific epitopes on CD110, for example, extracellular molecules of these antigens. Epitopes and other similar phenomena can be screened using these computational techniques. The identified antibody, its antigen-binding fragment, and ligand are used in the cancer treatment described herein. The treatment methods described herein, including methods for treating autoimmune diseases, and the treatment methods described herein. It can be used in conjunction with patient conditioning procedures.

[0233] CD45 on the surface of cells (e.g., cancer cells, autoimmune cells, or hematopoietic stem cells) , CD45RO), CD135, CD34, CD90, and / or CD110 bind, for example, receptor-mediated Antibodies taken up by cells through endocytosis, their antigen-binding fragments, and Further techniques can be used to identify ligands. For example, cancer cells, CD45 (e.g., CD45RO), CD135, CD34, CD9 on the surface of autoimmune cells or hematopoietic stem cells The antibody that binds to and / or CD110, and subsequently incorporates its antigen-binding fragment. Furthermore, to screen for ligands, the above in vitro display technique The technique can be applied. Phage display is a part of this screening paradigm. Represents one such technology that can be used in conjunction with the above targets. Next, antibodies are taken up by cancer cells, autoimmune cells, or hematopoietic stem cells, and their f To identify the ligand and ligand, those skilled in the art refer to Williams et al., Leukemia. The phage display technology described in 19:1432-1438, 2005 can be applied, and its development The whole of the references is incorporated herein by reference. For example, known in the art Using mutagenesis techniques, in particular antibodies, antibody fragments, such as scFv fragments, are used. T, Fab fragment, diabody, triabody, and 10 Fn3 domain, or Randomized amino acid cassette (e.g., one or more of the CDR or its equivalent regions, if A combination of ligands that contain all of the antibody or antibody fragments. A replacement phage library can be created. Framework region, hinge, Fc The main part, and other regions of the antibody or antibody fragment, for example, human germline antibodies. Having sequences that show only slight mutations relative to the body sequence or human germline antibodies Therefore, it can be designed to be non-immunogenic in humans.

[0234] Using phage display techniques described herein or known in the art Then, randomized antibodies, antibody fragments, or ligands covalently bound to phage particles are used. Phage libraries including CD45 (e.g., CD45RO), CD135, CD34, CD90, and / Alternatively, it can be incubated with CD110, for example, first, phage The library is blocked by a blocking agent (e.g., milk protein, bovine serum albumin, and / or An antibody that shows nonspecific protein binding when incubated with (or IgG, etc.), and its f A phage encoding a lagment or ligand, and an antibody or Remove the phage encoding that fragment, and then the phage library is hematopoietic. This is done by incubating the phage library with a population of stem cells. Incubate them for a sufficient amount of time with target cells such as autoimmune cells or hematopoietic stem cells. CD45-specific, CD135-specific, CD34-specific, CD90-specific, and / or CD110-specific antibodies, The antigen-binding fragment or ligand then binds to the cell surface antigen, and subsequently to cancer cells. , can be taken up by autoimmune cells or hematopoietic stem cells (e.g.) For example, 30 minutes to 6 hours at 4°C, or 1 hour at 4°C). Phages containing antibodies, their fragments, or ligands that do not exhibit sufficient affinity are Enables binding to and uptake by cancer cells, autoimmune cells, or hematopoietic stem cells. To achieve this, the cells are then washed with, for example, a cold (4°C) 0.1 M glycine buffer with a pH of 2.8. It can be removed by cancer cells, autoimmune cells, or hematopoietic stem cells. The injected antibody, its fragment, or phage bound to the ligand can, for example, enter a cell. Identifying the phages by dissolving them and recovering the incorporated phages from the cell culture medium. This can be done. Next, the phage can, for example, remove bacterial cells from the phage recovered in 2×YT medium. By incubating the bacteria using methods known in the art, bacterial cells can be released. It can be amplified in cells. The phage then recovered from this medium is, for example, f Genes encoding antibodies, their fragments, or ligands inserted into the phag genome. It can be characterized by determining the nucleic acid sequence of its offspring (one or more). The antibody, its fragment, or ligand is then (for example, scFV fragment) Chemical synthesis of antibody fragments such as ligands, or ligands (for example, full-length antibodies) It can be prepared de novo by recombinant expression.

[0235] The ability of the prepared antibody, its fragment, or ligand to be taken up is, for example, by the technique. This can be evaluated using radionuclide uptake assays known in the field. For example, If, then, an in vitro display described herein or known in the art Antibodies, their fragments, or ligands identified using the technology are 18 F, 75 Br, 7 7 Br, 122 I, 123 I, 124 I, 125 I, 129 I, 131 I, 211 At, 67 Ga, 111 In, 99 Tc, 169 Yb,186 Re , 64 Cu, 67 Cu, 177 Lu, 77 As, 72 As, 86 Y, 90 Y, 89 Zr, 212 Bi, 213 Bi, or 225 Business etc. Functionalization can be achieved by incorporating radioactive isotopes. For example, 18 F, 75 Br, 77 Br , 122 I, 123 I, 124 I, 125 I, 129 I, 131 I, 211 Radioactive halogens such as At are electrophilic halogens Gen reagents (e.g., iodized beads, Thermo Fisher Scientific, Ken) Using beads such as polystyrene beads containing bridge (Massachusetts), Antibodies, their fragments, or ligands can be incorporated. Radiolabeled antibodies, their fragments Ligands, or ligands, are taken up by cancer cells, autoimmune cells, or hematopoietic stem cells. Allow sufficient time for the ink to dry (for example, 30 minutes to 6 hours at 4°C, or 1 hour at 4°C). It can be converted. Next, the cells are washed to remove the antibodies that were not taken up, and their fu It can remove lagging or ligands (for example, 0.1M granules at pH 2.8, cold (4°C)). (Using lysine buffer) The incorporated antibody, its fragment, or ligand Radiation emitted from resulting cancer cells, autoimmune cells, or hematopoietic stem cells (e.g.) For example, comparing gamma rays with the radiation emitted from the recovered washing buffer (e.g., gamma rays). It can be identified by detection.

[0236] (Drug-antibody conjugates and drug-ligand conjugates) (Cytotoxin) Antibodies, antigen-binding fragments thereof, and ligands (e.g., CD45( Antibodies and antigens that recognize and bind to CD135, CD34, CD90, and / or CD110 (e.g., CD45RO), The binding fragment and ligand are, for example, Pseudomonas exotoxin A, deBouganin. Diphtheria toxin, amatoxins such as α-amanitin, saporins, maytansine, may Tansinoids, auristatin, anthracyclines, calicheamicin, irinotecan SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolobenzodiazepine (2 doses) The body, indolinobenzodiazepine, and indolinobenzodiazepine dimer, or Cytotoxins such as those variants, or known in this specification or in the art It can be conjugated to other cytotoxic compounds, for example, the purpose is For the treatment of cancer or autoimmune diseases as described in the specification, or for hematopoietic stem cell transplantation therapy. When administered to patients who require it (e.g., human patients), it promotes the depletion of endogenous hematopoietic stem cells. This is for the purpose of... In some embodiments, the cytotoxic molecule is an antibody, an antigen-binding fragment After the uptake of a ligand by the cell, the cytotoxin accesses its intracellular target and becomes endogenous Antibodies, their antigen-binding fragments, or that can be taken up in a way that can mediate hematopoietic cell death. It is conjugated to a ligand. It is used in conjunction with the compositions and methods described herein. Among the cytotoxins suitable for this field, DNA insertion agents are particularly well known. (e.g., anthracyclines), drugs that can disrupt the spindle apparatus (e.g., vinca) Alkaloids, maytansin, maytansinoids, and their derivatives), RNA polymers Ase inhibitors (e.g., amatoxins such as α-amanitin and their derivatives), Drugs that can disrupt protein biosynthesis (e.g., saporins and lysine A chains) Examples include drugs that exhibit RNA N-glycosidase activity.

[0237] In some embodiments, the cytotoxin is α-amanitin, β-amanitin, γ-amanitin , ε-amanitin, amanin, amaninamide, amanulin, amanulinic acid, and p Amatoxins or their derivatives, such as loamanulin. For example, as described herein. Antibodies, antigen-binding fragments, and ligands form a conjugate represented by the formula Ab-Am. It may bind to amatoxin to form a fragment, where Ab is the antibody and its antigen-binding fragment. Am is an amatoxin, or ligand. In some embodiments, Am is , represented by equation (I): [ka] [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4 is H, OH, OR C , ORD , R C , or R D and; R5 is H, OH, OR C , OR D , R C , or R D and; R6 is H, OH, OR C , OR D , R C , or R D and; R7 is H, OH, OR C , OR D , R C , or R D and; R8 is OH, NH2, OR C , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; and to Z is a reactive substituent present on L, and CD45 (CD45RO, etc.), CD135, CD34, CD90, and Present in the antibody that binds to CD110, its antigen-binding fragment, or its ligand. It is a chemical site formed by a coupling reaction with a reactive substituent.

[0238] In some embodiments, Am is just one R C Includes substituents.

[0239] In some embodiments, Am is represented by formula (IA): [ka] [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4 is H, OH, OR C , OR D , R C , or R D and; R5 is H, OH, OR C , OR D , R C , or R D and; R6 is H, OH, OR C , OR D , R C , or R D and; R7 is H, OH, OR C , OR D , R C , or R D and; R8 is OH, NH2, OR C , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; Z is a reactive substituent present on L, and CD45 (CD45RO, etc.), CD135, CD34, CD90, and Present in the antibody that binds to CD110, its antigen-binding fragment, or its ligand. It is a chemical site formed by a coupling reaction with a reactive substituent; and In the formula, Am is just one R C [Contains substituents].

[0240] In some embodiments, Am is represented by formula (IB): [ka] [In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R3 is H, R C , or R D and; R4 is H, OH, OR C , OR D , R C , or R D and; R5 is H, OH, OR C , OR D , R C , or R D and; R6 is H, OH, OR C , OR D , R C , or R D and; R7 is H, OH, OR C , OR D , R C , or R D and; R8 is OH, NH2, OR C , OR D NHR C , or NR C R D and; R9 is H, OH, OR C , or OR D and; X is -S-, -S(O)-, or -SO2-; R C is -LZ; R D This includes optionally substituted alkyl groups (e.g., C1-C6 alkyl groups) and optionally substituted heterozymes. C1-C6 heteroalkyls (e.g., C1-C6 heteroalkyls), optionally substituted alkenyls (e.g., C1-C6 heteroalkyls), 2-C6 alkenyls), optionally substituted heteroalkenyls (e.g., C2-C6 heteroalkenyls) (L), arbitrarily substituted alkynyl (e.g., C2-C6 alkynyl), arbitrarily substituted he Teloalkynyls (e.g., C2-C6 heteroalkynyls), optionally substituted cycloalkyls , optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally placed It is a converted heteroaryl; L is an arbitrarily substituted alkylene (e.g., C1-C6 alkylene), and an arbitrarily substituted he Teloalkylenes (e.g., C1-C6 heteroalkylenes), arbitrarily substituted alkenylenes ( For example, C2-C6 alkenylenes, or arbitrarily substituted heteroalkenylenes (for example, C2-C6 he Telalkenylenes), optionally substituted alkynylenes (e.g., C2-C6 alkynylenes), A heteroalkylene with arbitrary substitution (e.g., C2-C6 heteroalkylene), arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Linkers such as arylenes or optionally substituted heteroarylenes; Z is a reactive substituent present on L, and CD45 (CD45RO, etc.), CD135, CD34, CD90, and Present in the antibody that binds to CD110, its antigen-binding fragment, or its ligand. It is a chemical site formed by a coupling reaction with a reactive substituent; and In the formula, Am is just one R C [Contains substituents].

[0241] In some embodiments, R A and R B They are together with the oxygen atoms to which they are bonded. They then combine to form the following: [ka] [In the formula, Y is O, S, NR E , and CR E R E Selected from, and R E and R E' Each of these is independently and arbitrarily substituted C1-C6 alkylene-R C , replace as desired C1-C6 heteroalkylene-R C, optionally substituted C2-C6 alkenylene-R C , replace as desired C2-C6 heteroalkenylene-R C , optionally substituted C2-C6 alkynylene-R C , place as you like Replaced C2-C6 heteroalkylene-R C , optionally substituted cycloalkylene-R C , at will Substituted heterocycloalkylene-R C , optionally substituted arylene-R C , or optionally Substituted heteroarylene-R C It is.

[0242] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] R3 is H or R C and; R4 is H, OH, OR C , OR D , R C , or R D and; R5 is H, OH, OR C , OR D , R C , or R D and; R6 is H, OH, OR C , OR D , R C , or R D and; R7 is H, OH, OR C , OR D , R C , or R D and; R8 is OH, NH2, OR C , or NHR C and; R9 is either H or OH; and In the formula, R C and R D These are defined as described above.

[0243] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 is H, OH, OR A , or OR C and; R2 is H, OH, OR B , or OR C and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] R3 is H or R C and; R4 and R5 are independently H, OH, and OR, respectively. C , R C , or OR D and; R6 and R7 are H, respectively; R8 is OH, NH2, OR C , or NHR C and; R9 is either H or OH; and In the formula, R C This is as defined above.

[0244] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 is H, OH, or OR A and; R2 is H, OH, or OR B and; R A and R B These, together with the oxygen atoms to which they are bonded, combine to form the following: : [ka] R3, R4, R6, and R7 are each H; R5 is OR C and; R8 is either OH or NH2; R9 is either H or OH; and In the formula, R C This is as defined above. Such amatoxin conjugate is For example, it is described in U.S. Patent Application Publication No. 2016 / 0002298, and the disclosure is, in its entirety. This is incorporated herein by reference.

[0245] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 and R2 are independently either H or OH; R3 is R C and; R4, R6, and R7 are H, respectively; R5 is H, OH, or OC1-C6 alkyl; R8 is either OH or NH2; R9 is either H or OH; and In the formula, R C This is as defined above. Such amatoxin conjugate is For example, it is described in U.S. Patent Application Publication No. 2014 / 0294865, and the disclosure is, in its entirety. This is incorporated herein by reference.

[0246] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 and R2 are independently either H or OH; R3, R6, and R7 are each H; R4 and R5 are independently H, OH, and OR, respectively. C , or R C and; R8 is either OH or NH2; R9 is either H or OH; and In the formula, R C This is as defined above. Such amatoxin conjugate is For example, it is described in U.S. Patent Application Publication No. 2015 / 0218220, and the disclosure is, in its entirety. This is incorporated herein by reference.

[0247] In some embodiments, Am is represented by formula (IA) or formula (IB), In the formula, R1 and R2 are independently either H or OH; R3, R6, and R7 are each H; R4 and R5 are independently either H or OH; R8 is OH, NH2, OR C , or NHR C and; R9 is either H or OH; and In the formula, R C This is as defined above. Such amatoxin conjugate is For example, U.S. Patent Nos. 9,233,173 and 9,399,681, and U.S. Patent Application Publication No. 2016 This is described in publication / 0089450, and each of its disclosures is incorporated herein by reference in whole. It gets included.

[0248] According to the compositions and methods described herein, an antibody, its antigen-binding fragment, or Further amatoxins that can be used for ligand conjugation include, for example, WO2 As described in 016 / 142049;WO 2016 / 071856;and WO 2017 / 046658, each of these The disclosure is incorporated herein in its entirety by reference.

[0249] In some embodiments, Am is represented by equation (II): [ka] [wherein X is S, SO, or SO2; R1 is a linker covalently bound to H, or an antibody or its antigen-binding fragment. and; R2 is a linker covalently bound to H, or an antibody or its antigen-binding fragment. and; as well Here, if R1 is H, then R2 is a linker, and if R2 is H, then R1 is a linker. ru].

[0250] Antibodies and antigen-binding fragments for use in conjunction with the compositions and methods described herein. The ligands are conjugates known in the art or described herein. Using advanced technology, conjugate amatoxins such as α-amanitin or their variants. It can be used. For example, CD45 (CD45RO etc.), CD135, CD34, CD90, and / or Alternatively, an antibody that recognizes and binds to CD110, its antigen-binding fragment, and ligand are α- It can be conjugated to amatoxins such as amanitin or their variants, in the United States As described in Patent Application Publication No. 2015 / 0218220, the disclosure includes, for example, α-flaxseed. Amatoxins such as tin and their variants, as well as covalent conjugations, are used. As relating to a covalent linker that can perform the following, incorporated herein by reference It can be done.

[0251] Exemplary antibody-drug and ligand-drug conjugates useful in conjunction with the methods described herein. The gate connects an antibody, its antigen-binding fragment, or ligand to an antibody, its antigen-binding fragment Linkers containing substituents suitable for reaction with reactive residues on lgments or ligands. It can be formed by reaction with amatoxin conjugated to it. The anti substituents suitable for reaction with the body, its antigen-binding fragment, or reactive residues on the ligand. The following are examples of amatoxins that conjugate to linkers containing these Not limited to: 7'C-(4-(6-(maleimide)hexanoyl)piperazine-1-yl)-amatoxy n;7'C-(4-(6-(maleimido)hexaneamide)piperidine-1-yl)-amatoxin;7'C-(4 -(6-(6-(maleimide)hexanoyl)piperazine-1-yl)-amatoxin; 7'C-(4-(4-((maleimide)methyl)cyclohexanecarbonyl)piperazine-1-yl)-amato Xyn; 7'C-(4-(6-(4-((maleimide)methyl)cyclohexanecarboxamide)hexano (Lu)piperazine-1-yl)-amatoxin; 7'C-(4-(2-(6-(maleimide)hexanamide)eth (Lu)piperidine-1-yl)-amatoxin; 7'C-(4-(2-(6-(6-(maleimide)hexanamide) Hexaneamide)ethyl)piperidine-1-yl)-amatoxin; 7'C-(4-(2-(4-((maleimide Methyl(cyclohexanecarboxamide)ethyl(piperidine-1-yl)-amatoxin; 7'C -(4-(2-(6-(4-((maleimidomethyl)cyclohexanecarboxamide)hexaneamide) (Tyl)piperidine-1-yl)-amatoxin; 7'C-(4-(2-(3-carboxypropanamide)eth (Lu)piperidine-1-yl)-amatoxin; 7'C-(4-(2-(2-bromoacetamido)ethyl)piperidine Lysine-1-yl)-amatoxin; 7'C-(4-(2-(3-(pyridine-2-yldisulfanil)prop (Immido)ethyl)piperidine-1-yl)-amatoxin; 7'C-(4-(2-(4-(maleimide)butane (Amido)ethyl)piperidine-1-yl)-amatoxin; 7'C-(4-(2-(maleimide)acetyl)pi Perazin-1-yl)-amatoxin; 7'C-(4-(3-(maleimide)propanoyl)piperazine-1- 1-yl)-amatoxin; 7'C-(4-(4-(maleimide)butanoyl)piperazine-1-yl)-amatoxin Syn; 7'C-(4-(2-(6-(4-((maleimidomethyl)cyclohexanecarboxamide)hexane Amid-ethyl piperidine-1-yl amatoxin; 7'C-(3-((6-(maleimide)hexane Mido(methyl)pyrrolidine-1-yl)-amatoxin; 7'C-(3-((6-(6-(maleimide)hexane Amide)Hexaneamide)Methyl)Pyrrolidine-1-yl)-Amatoxin;7'C-(3-((4-((Mare Imido(methyl)cyclohexanecarboxamide(methyl)pyrrolidine-1-yl)-amatoxy 7'C-(3-((6-((4-(maleimidomethyl)cyclohexanecarboxamide)hexaneami (d)methyl)pyrrolidine-1-yl)-amatoxin; 7'C-(4-(2-(6-(2-(aminooxy)aceto Amide)Hexaneamide)Ethyl)Piperidin-1-yl)-Amatoxin;7'C-(4-(2-(4-(2-( Aminooxyacetamide butanamide ethyl piperidine-1-yl amatoxin; 7 'C-(4-(4-(2-(aminooxy)acetamide)butanoyl)piperazine-1-yl)-amatoxy N;7'C-(4-(6-(2-(aminooxy)acetamide)hexanoyl)piperazine-1-yl)-ama Toxin; 7'C-((4-(6-(maleimido)hexaneamide)piperidine-1-yl)methyl)-amato Xynxine; 7'C-((4-(2-(6-(maleimido)hexaneamide)ethyl)piperidine-1-yl)methyl )-Amatoxin; 7'C-((4-(6-(maleimide)hexanoyl)piperazine-1-yl)methyl)-A Matoxin; (R)-7'C-((3-((6-(maleimido)hexaneamide)methyl)pyrrolidine-1-yl) Methyl)-amatoxin; (S)-7'C-((3-((6-(maleimido)hexaneamide)methyl)pyrrolidine (1-yl)methyl)-amatoxin; 7'C-((4-(2-(6-(6-(maleimide)hexamide)hex Sanamido)ethyl)piperidine-1)yl)methyl)amatoxin; 7'C-((4-(2-(4-((Murray (Mido)methyl)cyclohexanecarboxamide)ethyl)piperidine-1-yl)methyl)-amato Xyn; 7'C-((4-(2-(6-(4-((maleimidomethyl)cyclohexanecarboxamide)hexa (Immido)ethyl)piperidine-1-yl)methyl)-amatoxin; 7'C-((4-(2-(6-(Malaymi (Hexaneamide)ethyl)piperazine-1-yl)methyl)-amatoxin; 7'C-((4-(2-(6-( 6-(maleimide)hexaneamide)hexaneamide)ethyl)piperazine-1-yl)methyl)- Matoxin; 7'C-((4-(2-(4-((maleimide)methyl)cyclohexanecarboxamide)eth (L)piperazine-1-yl)methyl)-amatoxin; 7'C-((4-(2-(6-(4-((maleimide)methyl) Cyclohexanecarboxamide (hexaneamide) ethyl (piperazine-1-yl) methyl (a) Matoxin; 7'C-((3-((6-(6-(maleimido)hexaneamide)hexaneamide)-S-methyl) Pyrrolidine-1-yl)methyl)-amatoxin; 7'C-((3-((6-(6-(maleimide)hexaneamide (d)hexanamide)-R-methyl)pyrrolidine-1-yl)methyl)-amatoxin; 7'C-((3-((4- ((maleimide)methyl)cyclohexanecarboxamide)-S-methyl)pyrrolidine-1-yl) Chil)-Amatoxin; 7'C-((3-((4-((maleimide)methyl)cyclohexanecarboxamide )-R-methyl)pyrrolidine-1-yl)methyl)amatoxin; 7'C-((3-((6-(4-((maleimide) Methyl)cyclohexanecarboxamide)hexanamide)methyl)pyrrolidine-1-yl)meth (Lu)-Amatoxin; 7'C-((4-(2-(3-Carboxypropanamide)ethyl)piperazine-1-I (L)methyl)-amatoxin; 7'C-((4-(6-(6-(maleimide)hexanoyl) Piperazine-1-yl)methyl)-amatoxin; 7'C-((4-(6-(4-((maleimide)methyl)cyclo Hexanecarboxamide (hexanoyl)piperazine-1-yl)methyl)-amatoxin; 7'C- ((4-(2-(maleimide)acetyl)piperazine-1-yl)methyl)-amatoxin; 7'C-((4-(3-( Maleimide propanoyl piperazine-1-yl methyl amatoxin; 7'C-((4-(4-(Maleimide) Imido)butanoyl)piperazin-1-yl)methyl)amatoxin; 7'C-((4-(2-(2-(Murray (Mido)acetamido)ethyl)piperidine-1-yl)methyl)-amatoxin; 7'C-((4-(2-(4-( Maleimide (butanamide) ethyl (piperidine-1-yl) methyl (amatoxin) 7'C-((4-( 2-(6-(4-((maleimidomethyl)cyclohexanecarboxamide)hexaneamide)ethyl) Piperidine-1-yl)methyl)-amatoxin; 7'C-((3-((6-(maleimide)hexanamide) Methyl)azetidine-1-yl)methyl)amatoxin; 7'C-((3-(2-(6-(maleimide)hexa (Ammido)ethyl)azetidine-1-yl)methyl)-amatoxin; 7'C-((3-((4-((maleimide Methyl)cyclohexanecarboxamide)methyl)azetidine-1-yl)methyl)-amatoxy N; 7'C-((3-(2-(4-((maleimide)methyl)cyclohexanecarboxamide)ethyl)azeti Zin-1-yl)methyl)-amatoxin; 7'C-((3-(2-(6-(4-((maleimide)methyl)cyclohex Sancarboxamide (Hexaneamide) Ethyl (Azetidine-1-yl) Methyl (Amatoxin) ;7'C-(((2-(6-(maleimido)-N-methylhexaneamide)ethyl)(methyl)amino)methyl)- Amatoxin; 7'C-(((4-(6-(maleimide)N-methylhexaneamide)butyl(methyl)amide (no)methyl)-amatoxin; 7'C-((2-(2-(6-(maleimido)hexanamide)ethyl)azili Zin-1-yl)methyl)-amatoxin; 7'C-((2-(2-(6-(4-((maleimide)methyl)cyclohex Xancarboxamide (hexaneamide) ethyl (aziridin-1-yl) methyl (amatoxy) n;7'C-((4-(6-(6-(2-(aminooxy)acetamide)hexanoyl)pipette Radin-1-yl)methyl)-amatoxin; 7'C-((4-(1-(aminooxy)-2-oxo-6,9,12,15 -Tetraoxa-3-azaheptadecan-17-oil)piperazine-1-yl)methyl)amatoxyl n;7'C-((4-(2-(2-(aminooxy)acetamide)acetyl)piperazine-1-yl)methyl)- Amatoxin; 7'C-((4-(3-(2-(aminooxy)acetamide)propanoyl)piperazine-1 -yl)methyl)-amatoxin; 7'C-((4-(4-(2-(aminooxy)acetamide)butanoyl) Piperazine-1-yl(methyl)-amatoxin; 7'C-((4-(2-(6-(2-(aminooxy)aceta Mido)hexanamide)ethyl)piperidine-1-yl)methyl)-amatoxin; 7'C-((4-(2-(2 -(2-(aminooxy)acetamide)acetamide)ethyl)piperidine-1-yl)methyl)- Matoxin; 7'C-((4-(2-(4-(2-((aminooxy)acetamide)butanamide)ethyl)pipette Lysine-1-yl)methyl)-amatoxin; 7'C-((4-(20-(aminooxy)-4,19-dioxo-6,9 ,12,15-tetraoxa-3,18-diazycosyl)piperidine-1-yl)methyl)-amatoxin; 7'C-(((2-(6-(2-(aminooxy)acetamide)-N-methylhexaneamide)ethyl)(methyl )amino)methyl)-amatoxin; 7'C-(((4-(6-(2-(aminooxy)acetamide)-N-meth (Xenamide)butyl)(methyl)amino)methyl)-amatoxin; 7'C-((3-((6-(4-((Ma Reimide)methyl)cyclohexanecarboxamide)hexaneamide)methyl)pyrrolidine-1- Iyl)-S-methyl)-amatoxin; 7'C-((3-((6-(4-((maleimide)methyl)cyclohexane Carboxamide (hexaneamide)-R-methyl)pyrrolidine-1-yl)methyl)-amatoxin; 7'C-((4-(2-(2-bromoacetamidoethyl)piperazine-1-yl)methyl)amatoxin; 7'C-((4-(2-(2-bromoacetamidoethyl)piperidine-1-yl)methyl)amatoxin; 7'C-((4-(2-(3-(pyridine-2-yldisulfanyl)propanamide)ethyl)piperidine-1- (Iyl)methyl)-amatoxin; 6'O-(6-(6-(maleimide)hexaneamide)hexyl)-amato Xyn; 6'O-(5-(4-((maleimide)methyl)cyclohexanecarboxamide)pentyl)-A Matoxin; 6'O-(2-((6-(maleimide)hexyl)oxy)-2-oxoethyl)-amatoxin ;6'O-((6-(maleimide)hexyl)carbamoyl)-amatoxin;6'O-((6-(4-((maleimide) Methyl(cyclohexanecarboxamide)hexyl(carbamoyl)-amatoxin; 6'O- (6-(2-bromoacetamide)hexyl)-amatoxin; 7'C-(4-(6-(azido)hexaneamide (d)piperidine-1-yl)-amatoxin; 7'C-(4-(hexa-5-inoylamino)piperidine-1 -yl)-amatoxin; 7'C-(4-(2-(6-(maleimido)hexanamide)ethyl)piperazine-1 -yl)-amatoxin; 7'C-(4-(2-(6-(6-(maleimido)hexaneamide)hexaneamide) Ethyl)piperazin-1-yl)-amatoxin; 6'O-(6-(6-(11,12-didehydro-5,6-dihydro -Dibenzo[b,f]azosin-5-yl)-6-oxohexaneamide)hexyl)-amatoxin;6' O-(6-(hexa-5-inoylamino)hexyl)-amatoxin; 6'O-(6-(2-(aminooxy)a Cetylamide)hexyl)-amatoxin; 6'O-((6-aminooxy)hexyl)-amatoxin ; and 6'O-(6-(2-iodoacetamide)hexyl)-amatoxin. The aforementioned linker, In particular, it is useful in conjunction with the compositions and methods described herein, for example, in a U.S. patent application. This information is contained in Publication No. 2015 / 0218220, and its entirety is referred to herein by reference. It will be incorporated.

[0252] For use in the direct treatment of cancer and autoimmune conditions, or in preparation for hematopoietic stem cell transplantation. CD45 (CD45RO, etc.), CD for conditioning patients (e.g., human patients) Antibodies that recognize and bind to CD135, CD34, CD90, and / or CD110, and their antigen-binding fragments Further cytotoxins that can be conjugated to ligands include, among others, the following: These are some, but are not limited to: 5-ethinyluracil, abiraterone, acylflube N, adesipenolic acid, adzeresin, aldresleukin, altretamine, ambamust N, Amidox, Amiphostin, Aminolevulinic Acid, Amrubicin, Amsacrin, A Nagrelide, anastrozole, andrografolide, angiogenesis inhibitors, Antarelic S, anti-dorsal morphogenesis protein-1, anti-androgen, prostate cancer, anti-estrogen, anti New biopharmaceuticals, antisense oligonucleotides, aphydicolinglycinate, apotosis gene modulator, apoptosis regulator, aprinic acid, asracrin, Tamestane, atrimustine, axinastatin 1, axinastatin 2, axinastatin N3, Azasetron, Azatoxin, Azatyrosine, Baccatin III derivative, Valanol, Batimastat, BCR / ABL antagonist, benzochlorine, benzoyl staurospoly N, beta-lactam derivatives, beta-aretin, betacramycin B, betulinic acid, bFGF inhibitor Harmful agents, bicalutamide, bisanthren, bisaziridinylspermine, bisnafid, bis Traten A, Bizeresin, Blefrate, Bleomycin A2, Bleomycin B2, Bropi Limin, Budotitan, Butionine Sulfoximine, Calcipotriol, Carphostin C Camptothecin derivatives (e.g., 10-hydroxy-camptothecin), capecitabine, Ruboxamide-amino-triazole, carboxamide triazole, carzeresin, Zein kinase inhibitors, castanospermine, cecropin B, cetrorelix, chloride Chloroquinoxaline sulfonamide, cicaprost, cis-porphyrin, cladyl Bin, clomiphene and its analogues, clotrimazole, colismycin A, colisma Icin B, Combretastatin A4, Combretastatin analog, Conagenin, Crambeth Zin 816, Crisnator, Cryptophycin 8, Cryptophycin A derivative, Curacin A Cyclopentaanthraquinone, cycloplatam, cypemycin, cytarabine ocphosph Glutamate, cytolytic factors, cytostatin, dacliximab, decitabine, dehydrosidem Nin B, 2'-deoxycoformycin (DCF), deslorerin, dexyphosphamide, dex Slazoxane, dexverapamil, diazicon, didemnin B, zidox, diethylno Luspermine, dihydro-5-azacitidine, dihydrotaxol, dioxamycin, di Phenylspiromustine, Discodermolide, Docosanor, Dracetron, Doxyf Lurizine, droloxifen, dronabinol, duocalmycin SA, ebselen, e Comstin, Edelfosine, Edrecolomab, Eflornithine, Elemen, Emiteflu Epothiron, Epitiron, Epristeride, Estramustine and its analogs, Eto Poside, etoposide 4'-phosphate (also called etopophos), exemestane, fad Rozole, Fazarabine, Fenretinide, Filgrastim, Finasteride, Flavo Pyridol, Frezelastine, Fluasterone, Fludarabine, Fluorodaurine hydrochloride Syn, Holphenimex, Formestan, Fostoriesin, Fotemstin, Gad Linium texaphylline, gallium nitrate, gallocitabine, ganirelix, gelatinase Inhibitors, gemcitabine, glutathione inhibitors, hepsulfame, homohalinguin (HH T) Hypericin, ibandronate, idoxifen, idramanthon, irmofosin, Ilostat, imidazoacridone, imiquimod, immunostimulatory peptide, yobengguan Iododoxorubicin, Ipomeanol, Irinotecan, Iroprocto, Ilsoglaz N, isobengazole, jasplaquinolide, kahalalide F, lamelin-N triacetate T, Lanreotide, Reinamycin, Renograstim, Lentinan sulfate, Leptol state Letrozole, lipophilic platinum compound lysocrineamide 7, lobaplatin, lometrexo Lul, ronidamin, loxoxantrone, loxoribine, lulutotecan, lutetium tequil Saffirine, Lysophylline, Masopropyl, Maspin, Matrix Metalloprotein Methionase inhibitors, menogalil, lunervalon, metheterine, methioninase, metoclopramide MIF inhibitors, ifepristone, miltefosine, mirimostim, mitracin, mito Guazone, Mitractol, Mitomycin and its analogs, Mitonafide, Mitoxa Ntron, mophalotene, moglamostin, micaperoxide B, miriapolon, N- Acetyl dinarin, N-substituted benzamide, nafarelin, nagressip, napabine, na Phterpine, naltograstim, nedaplatin, nemorubicin, neridronic acid, niltami D, Nisamycin, Nitrulline, Octreotide, Oxenon, Onapristone, Onda Nsetron, Oracin, Ormaplatin, Oxaliplatin, Oxaunomycin, Pa Ritaxel and its analogues, paraamines, palmitoyl lyzoxin, pamidronic acid, Panaxytriol, Panomiphene, Parabactin, Pazeliptin, Pegaspargase, Perdesine, Pentosan polysulfate sodium, Pentostatin, Pentrozole, Perf Lubron, perphosphamide, phenazinomycin, picibanil, pirarubicin, pyri Torexim, podophyllotoxin, porphyromycin, purine nucleoside phosphorylates -ase inhibitors, larcitrexed, lyzoxin, logretimide, rohypnol, rubiginone B1 Ruboxil, Safinol, Saintpin, Sarcophytol A, Sarglamostim Sobuzoxane, Sonelmin, Sparfosinate, Spicamycin D, Spiromustine, S Tipiamid, sulfinosine, talimustine, tegafur, temozolomide, teniposide, Talibrastin, Thiocoralin, Tirapazamin, Topotecan, Topsentin, Trisilib Trimethrexate, Bellamine, Vinorelbine, Binxaltin, Borozol, Zeni Platin, and also Girascorb.

[0253] (Linker for chemical conjugation) Using various linkers, the antibodies, antigen-binding fragments, and and ligands (e.g., CD45 (CD45RO, etc.), CD135, CD34, CD90, and / or CD110) Antibodies that recognize and bind to antigens, their antigen-binding fragments, and ligands (which are cytotoxic components) It can be conjugated with its offspring. The linker can be subjected to, for example, enzymatic hydrolysis and photodegradation. Hydrolysis under acidic conditions, hydrolysis under basic conditions, oxidation, disulfide reduction, nucleophilicity Examples include those that can be cleaved by cleavage or organometallic cleavage (e.g., Leriche et al.) See al., Bioorg. Med. Chem., 20:571-582, 2012, the disclosure of which is a covalent conjugate. (This is incorporated herein by reference as relating to linkers suitable for gates.) Linkers useful for the synthesis of drug-antibody conjugates and drug-ligand conjugates Examples include, in particular, Michael receptors (e.g., maleimide), activated esters, and electron deficiencies. Carbonyl compounds and aldehydes, among others, are present within antibody or antigen-binding fragments. It contains an electrophile suitable for reaction with nucleophilic substituents such as amine or thiol moieties. This includes things like drug-antibody conjugates and drug-ligand conjugates. Suitable linkers for the synthesis of are not limited to, but in particular, 4-(N-maleimidomethyl) -Cyclohexane-L-carboxylic acid succinimidyl (SMCC), N-succinimidyliodoure Setate (SIA), sulfo-SMCC, m-maleimidobenzoyl-N-hydroxysuccinimid Examples include luesters (MBS), sulfo-MBS, and succinimidyliodoacetate. For example, it is described in Liu et al., 18:690-697, 1979, and its disclosure is chemical condyloma. The linker for linkage is incorporated herein by reference. The linker is particularly useful for the conjugation of microtubule disruptors such as auristatin. One example is a non-cleaving maleimidocaproyl linker, as described by Doronina et al., Bioconjugate. It is described in e Chem. 17:14-24, 2006, and the disclosure is for chemical conjugation. The following are incorporated herein by reference as relating to linkers. - Additional linker suitable for the synthesis of antibody conjugates and drug-ligand conjugates Some can release cytotoxins through the 1,6-elimination process, such as p-amines. Nobenzyl alcohol (PABC), 6-maleimidohexanoic acid, pH-sensitive carbonate, and Jain Other reagents are listed in et al., Pharm. Res. 32:3526-3540, 2015, and their disclosure is as follows: The entirety of that is incorporated herein by reference.

[0254] Conjugate antibodies, their antigen-binding fragments, or ligands to cytotoxic agents. Linkers that can be used for this purpose include one that is covalently bound to a cytotoxic agent at one end of the linker. And, at the other end of the linker, the reactive substituent present on the linker and CD45 (CD45RO, etc.), Antibodies that bind to CD135, CD34, CD90, and / or CD110, their antigen-binding fragments, Alternatively, the chemicals formed from coupling reactions between reactive substituents present in the ligand and the ligand. Examples include those containing the target part. CD45 (CD45RO, etc.), CD135, CD34, CD90, and / or present within the antibody that binds to CD110, its antigen-binding fragment, or the ligand. The reactive substituents that can be obtained include, but are not limited to, serine and threonine. , and the hydroxyl portion of the tyrosine residue; the amino portion of the lysine residue; aspartic acid and the carboxyl portion of glutamic acid residues; and the thiol portion of cysteine ​​residues, In addition, propargyl, azide, haloaryl (e.g., fluoroaryl), haloheterol Reels (e.g., fluoroheteroaryls), haloalkyls, and naturally occurring compounds Haloheteroalkyl moiety of amino acids. Antibody-drug and ligand-drug compounds as described herein. Linkers useful in combination with jugates include coupling reactions, as shown in Table 2 below. This includes, but is not limited to, linkers containing chemical sites formed by the reaction. The curves represent the binding points to antibodies, antigen-binding fragments, or ligands and cytotoxic molecules. These are shown below.

[0255] Coupling reactions in the formation of antibody-drug and ligand-drug conjugates Exemplary chemical sites formed [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5]

[0256] (Treatment method) As described herein, hematopoietic stem cell transplantation therapy is administered to subjects in need of treatment. It can be applied to proliferate or regrow one or more blood cell types. Hematopoietic stem cells are They usually exhibit pluripotency, and therefore granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils) ), red blood cells (e.g., reticulocytes, red blood cells), platelets (e.g., megakaryocytes, platelet-producing meganuclei) Spheres (platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglial cells, osteoclasts , and lymphocytes (e.g., NK cells, B cells, and T cells), but not limited to these. Hematopoietic stem cells can differentiate into multiple different bloodlines. Therefore, it is possible to produce daughter cells that have the same potential as the parent cells, and transplantation They are reintroduced into the cypient, where they home to the hematopoietic stem cell niche, and productively It is also characterized by its ability to restore sustained hematopoiesis.

[0257] Therefore, in order to reconstruct a defective or deficient cell population in vivo, hematopoietic stem cells The cells can be administered to patients who have defects or deficiencies in one or more cell types of the hematopoietic lineage. This treats conditions associated with defects or deficiencies in the endogenous blood cell population. The compositions and methods described herein are for non-malignant abnormal hemoglobin disorders (e.g., sickle cell anemia). Celluloid anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiscott-Aldori anemia. It can be used to treat abnormal hemoglobin disorders (selected from a group including tsch syndrome). Additionally or alternatively, the compositions and methods described herein may be used for congenital immunodeficiency, etc. It can be used to treat immunodeficiency disorders. Additionally or alternatively, as specified herein. The compositions and methods described herein are for the treatment of acquired immunodeficiency (e.g., the group consisting of HIV and AIDS). It can be used to treat selected acquired immunodeficiency disorders. The compositions and methods described herein are for treating metabolic disorders (e.g., glycogen storage disease, mucopolysaccharidosis, gout disease). It consists of E. leukodystrophy, Hurler's disease, sphingolipidosis, and metachromatic leukodystrophy. It can be used to treat metabolic disorders (selected from the group).

[0258] Additionally or alternatively, the compositions and methods described herein are for the treatment of hematological cancers, myeloproliferative cancers. It can be used to treat malignant tumors or proliferative disorders such as cancer. In this case, the endogenous hematopoietic stem cell population is depleted before hematopoietic stem cell transplantation, as specified in this document. The compositions and methods described in this book can be applied to patients, in which case the transplanted cells will be endogenous. The sex cell depletion process allows for homing into the niche created, establishing productive hematopoiesis. This can then be done to deplete cells during cancer cell eradication, such as during systemic chemotherapy. The population can be reconstructed. Treatment using the compositions and methods described herein. Exemplary blood cancers that can be diagnosed include acute myeloid leukemia, acute lymphoblastic leukemia, and chronic myeloid leukemia. Chronic lymphocytic leukemia, multiple myeloma, diffuse large B-cell lymphoma, and This includes non-Hodgkin lymphoma and other cancerous conditions, including neuroblastoma. These are not the only options.

[0259] Further diseases that can be treated with the compositions and methods described herein include adeno Synthetic deficiency and severe combined immunodeficiency, hyperimmune globulin M syndrome, Chedi Ack-Dong syndrome, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, saccharomyces, Racemy major, systemic sclerosis, systemic lupus erythematosus, multiple sclerosis, and juvenile Rheumatoid arthritis is one example, but it is not limited to these.

[0260] Antibodies, antigen-binding fragments, ligands, and conjugates described herein. These can be used to induce solid organ transplant tolerance. For example, the compositions described herein. and methods for depleting or removing cell populations from target tissue (e.g., bone marrow stem cells) It can be used to deplete hematopoietic stem cells from a niche. Following the depletion of cells, a population of stem cells or progenitor cells from an organ donor (e.g., organ donor) Hematopoietic stem cells (from [source]) are administered to the transplant recipient, and the life of such stem cells or progenitor cells is restored. Following this, a temporary or stable mixed chimera can be achieved, thereby further This enables long-term transplant organ resistance without the need for immunosuppressants. For example, as described herein. The composition and method are for solid organ transplant recipients (e.g., kidney transplants, lung transplants, etc.). It can be used to induce transplant tolerance in liver transplants and heart transplants. The compositions and methods described in this book are, for example, for low proportions of temporary or stable donor grafts. Because it is sufficient to induce long-term resistance in transplanted organs, it is related to the induction of solid organ transplant resistance. It is well suitable for use.

[0261] Furthermore, the compositions and methods described herein include CD45+, CD135+, CD34+, CD90+, and It can be used to directly treat cancers such as those characterized by CD110+ cells. For example, the compositions and methods described herein are particularly effective for CD45+, CD135+, CD34+, and CD90 To be used to treat leukemia in patients showing CD110+ or ​​CD110+ leukemia cells. This can be done. It can convert CD45+, CD135+, CD34+, CD90+, or CD110+ cancer cells such as leukemia cells. By depleting, the compositions and methods described herein directly cause various cancers It can be used for treatment. Examples of cancers that can be treated in this way include acute myeloid leukemia. Acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, diffuse Examples of blood cancers include large B-cell lymphoma and non-Hodgkin lymphoma.

[0262] Furthermore, the compositions and methods described herein may be used to treat autoimmune diseases. It is possible to kill, for example, CD45+, CD135+, CD34+, CD90+, or CD110+ immune cells. In this way, antibodies, their antigen-binding fragments, or ligands are used in patients suffering from autoimmune diseases. It can be administered to human patients and other subjects. CD45+, CD135+, CD34+, CD90+, CD110+ immune cells specifically bind to autoantigens and trigger an immune response against them. These may be autoreactive lymphocytes such as T cells that express cytoreceptors. Autoreactive, CD45+, By depleting CD135+, CD34+, CD90+, or CD110+ cells, as described herein The compositions and methods are used to treat autoimmune conditions such as those described below. It may be. Additionally or alternatively, the compositions and methods described herein may be used for hematopoietic stem cell transplantation. To treat autoimmune diseases by depleting the population of endogenous hematopoietic stem cells before treatment. It can be used for this purpose, in which case the transplanted cells are placed in the niche created by the endogenous cell depletion process. It can be suppressed and productive hematopoiesis can be established. This, in turn, can be autoimmune It is possible to reconstruct the depleted cell population during cell eradication.

[0263] Autoimmune diseases that can be treated using the compositions and methods described herein include These include, but are not limited to, psoriasis, psoriatic arthritis, and type 1 diabetes mellitus. (Type 1 diabetes), rheumatoid arthritis (RA), systemic lupus leukemia (SLE), multiple sclerosis (MS), inflammation Intestinal disease (IBD), lymphocytic colitis, acute disseminated encephalomyelitis (ADEM), Addison's disease, all Alopecia, ankylosing spondylitis, antiphospholipid syndrome (APS), aplastic anemia, autoimmune disease hemolytic anemia, autoimmune hepatitis, autoimmune otitis interna (AIED), autoimmune lymphoproliferative disease ALPS syndrome, autoimmune oophoritis, Barlow's disease, Behçet's disease, bullous pemphigoid, cardiomyopathy Chagas disease, chronic fatigue immunodeficiency syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, chloro Celiac disease, pemphigoid scarring, celiac disease - herpetiform dermatitis, cold agglutinin disease, CREST syndrome, Gor's disease, lupus discoid, autonomic nervous system dysfunction, endometriosis, essential mixed cryoglobulinemia Fibromyalgia - Fibromyitis, Goodpasture syndrome, Graves' disease, Guillain-Barré syndrome (GBS), Hashimoto's thyroiditis, hidradenitis suppurativa, idiopathic and / or acute thrombocytopenic purpura , idiopathic pulmonary fibrosis, IgA neuropathy, interstitial cystitis, juvenile arthritis, Kawasaki disease, lichen planus, rheumatoid arthritis Imm's disease, Meniere's disease, mixed connective tissue disease (MCTD), myasthenia gravis, neuromuscular dystrophy, O Psoclonus-myoclonus syndrome (OMS), optic neuritis, oat thyroiditis, thyroiditis vulgaris Smallpox, pernicious anemia, polychondritis, polymyositis and dermatomyositis, primary biliary cirrhosis, nodules Polyarteritis genitourin, polyglandular syndrome, polymyalgia rheumatica, primary agammaglobulinemia, Ray No phenomenon, Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjögren's syndrome Group, Stiff Person syndrome, Takayasu's arteritis, Temporal arteritis (also known as "giant cell arteritis") (The following conditions are listed): ulcerative colitis, collagenous colitis, uveitis, vasculitis, vitiligo, vulvodynia. ("Vulvovaginitis"), and Wegener's granulomatosis.

[0264] (Route of administration and dosage) The antibodies, antigen-binding fragments, and ligands described herein are used in patients (e.g., Humans suffering from cancer, autoimmune diseases, or those requiring hematopoietic stem cell transplantation. It can be administered to patients in various dosage forms. For example, the antibody described herein, Antigen-binding fragments and ligands are used in patients with cancer, autoimmune diseases, and Alternatively, for patients requiring hematopoietic stem cell transplantation, one or more pharmaceutically acceptable excipients are included. It can be administered in the form of an aqueous solution, such as an aqueous solution containing the following. Pharmaceutically acceptable excipients used in conjunction with the method include viscosity modifiers. The solution can be sterilized using techniques known in the art.

[0265] The antibodies, antigen-binding fragments, and ligands described herein may be administered orally, transdermally, or subcutaneously. It can be administered via various routes, including intranasal, intravenous, intramuscular, intraocular, or parenteral administration. Yes, it is possible. The most suitable route of administration in any given case is the route of administration to the specific antibody, anti Original binding fragment or ligand, patient, pharmaceutical formulation method, method of administration (e.g., at the time of administration) (Interval and route of administration), patient's age, weight, sex, severity of the disease being treated, patient's diet It depends on the circumstances and the patient's excretion rate.

[0266] The effective amount of the antibody, its antigen-binding fragment, or ligand described herein is, for example, For single (e.g., bolus) administration, multiple administrations, or consecutive administrations, the amount is approximately 0.001 to approximately 1 It may be in the range of 00 mg / kg body weight, or the antibody, its antigen-binding fragment, or Within the range that achieves the optimal serum concentration of the ligand (for example, a serum concentration of 0.0001 to 5000 μg / mL) It is possible. The dosage is for subjects with cancer, autoimmune diseases (e.g., humans), or artificial. In subjects undergoing conditioning therapy in preparation for hematopoietic stem cell transplantation, 1 It may be administered once or more times a day, week, or month (for example, 2 to 10 times). In the case of conditioning procedures before cyst transplantation, antibodies, their antigen-binding fragments, or Gando is used to optimally promote the engraftment of exogenous hematopoietic stem cells in patients, for example, exogenous hematopoietic stem cells 1 hour to 1 week after administration of hematopoietic stem cell graft (e.g., 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours) Hours: 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours Intervals: 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 It can be administered on the 5th, 6th, or 7th day or earlier. [Examples]

[0267] The following examples illustrate how the compositions and methods described herein are used and manufactured. To provide those skilled in the art with an explanation of how it can be evaluated, purely illustrative and The intention is to limit the scope of what inventors consider to be their inventions. Not illustrated.

[0268] (Example 1: Administration of anti-CD45 antibody to human patients in preparation for hematopoietic stem cell transplantation) According to the method disclosed herein, exogenous hematopoietic stem cell grafts are used prior to hematopoietic stem cell transplantation therapy. In order to promote engraftment, internists in this profession condition patients such as human patients. This can be done. For this purpose, internists in this profession use antigens expressed by hematopoietic stem cells. An antibody or its antigen-binding fragment that can bind, for example, an antibody that binds to CD45. or its antigen-binding fragment (e.g., an antibody that binds to CD45RO or its antigen-binding fragment) Antibodies such as lagment can be administered to human patients. Antibodies are as described in this specification. Covalently conjugates to toxins such as cytotoxic molecules known in the art, or toxins. It can be gated. For example, an anti-CD45 antibody or its antigen-binding fragment (anti-CD45 antibody). 45RO antibody or its antigen-binding fragment, etc., for Pseudomonas exotoxin A, deBougani n, diphtheria toxin, amatoxins such as α-amanitin, saporin, maytansine, may Tansinoids, auristatin, anthracyclines, calicheamicin, irinotecan SN-38, Duocalmycin, Pyrrolobenzodiazepine, Pyrrolobenzodiazepine (2 doses) indolinobenzodiazepines, indolinobenzodiazepine dimers, or their bodies It can be covalently conjugated to cytotoxins such as mutants. The combination is a covalent bond described herein or known in the art. This can be carried out using manufacturing techniques. Antibodies, their antigen-binding fragments, or drugs -The antibody conjugate is then administered to the patient, for example, intravenously, to stimulate the extrinsic hematopoietic stem cells. It can be administered before transplantation to patients receiving autologous, allogeneic, or allogeneic hematopoietic stem cells. ru.

[0269] The amount of endogenous hematopoietic stem cells is reduced before hematopoietic stem cell transplantation, for example, to 10%, 20%, 30%, 40%, 50%. A sufficient amount of anti-CD45 to reduce by %, 60%, 70%, 80%, 90%, 95%, or more (for example) , administer an anti-CD45RO antibody, its antigen-binding fragment, or a drug-antibody conjugate. It is possible to reduce the number of hematopoietic stem cells using conventional techniques known in the field. For example, blood samples taken from the patient at various intervals during conditioning therapy. This can be monitored by FACS analysis of cells expressing characteristic hematopoietic stem cell surface antigens. This is possible. For example, an internist in this profession can assess the patient at various points during conditioning therapy. Blood samples can be collected from individuals, and FACS analysis can be performed to determine the relative concentration of hematopoietic stem cells in the samples. By elucidating the degree using antibodies that bind to hematopoietic stem cell marker antigens, we can determine the extent of endogenous hematopoiesis. The degree of stem cell reduction can be determined. According to some embodiments, hematopoietic stem cells The concentration of anti-CD45 (e.g., anti-CD45RO) antibody, its antigen-binding fragment, or drug-anti When the minimum value is reached in response to conditioning therapy with a body conjugate, the physician Even after completing conditioning therapy and beginning preparations for hematopoietic stem cell transplantation, good.

[0270] Anti-CD45 (e.g., anti-CD45RO) antibody, its antigen-binding fragment, or drug-antibody compound A solution of jugate containing one or more pharmaceutically acceptable excipients, such as viscosity modifiers. It can be administered to patients. The aqueous solution is as described herein or in the relevant technology. Sterilization can be performed using techniques known in the field. Hematopoietic stem cell grafts are administered to patients. Before administering, the antibody, its antigen-binding fragment, or drug-antibody conjugate, for example, For example, it can be administered to patients at doses ranging from 0.001 mg / kg to 100 mg / kg. Antibodies, their antigenic antibodies The combined fragment, or drug-antibody conjugate, is used to engraft exogenous hematopoietic stem cells in patients. At the point in time that best promotes this, for example, 1 hour to 1 week after administration of exogenous hematopoietic stem cell graft (for example) ba, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 o'clock Intervals: 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours (22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days) or longer It can be administered beforehand.

[0271] After the completion of the conditioning therapy, the patient then underwent further conditioning therapy. Injection of exogenous hematopoietic stem cells (e.g., intravenously) from the same internist or from a different internist. Intravenous injection is available. The physician can administer autologous, allogeneic, or allogeneic hematopoietic stem cells. For example, the injection is 1x10 3 ~1x10 9 Hematopoietic stem cells can be administered to patients at a dose of one kilogram. For example, an internist might take a blood sample from a patient and examine hematopoietic stem cells or cells of the hematopoietic lineage (meganuclei). Spheres, platelets, platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglia Cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural cells Measuring the increase in the concentration of grafts (such as Lar cells, T lymphocytes, and B lymphocytes) after administration. This allows us to monitor the engraftment of hematopoietic stem cell grafts. This analysis, for example, For example, it can be performed for 1 hour to 6 months or more after hematopoietic stem cell transplantation treatment. ba, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 o'clock Intervals: 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours Interval, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks Between 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 Weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 (weeks or longer). The concentration of hematopoietic stem cells or hematopoietic lineage cells is appropriate for the period prior to transplantation. Compared to the concentration of the cell type after transplantation (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%) 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, or any of the above. The above findings indicate an increase in anti-CD45 (e.g., anti-CD45RO) antibodies, and their antigen-binding fragments. Treatment with a drug-antibody conjugate improves the engraftment of transplanted hematopoietic stem cell grafts. This serves as one indicator that it was successfully promoted.

[0272] (Example 2: Administration of anti-CD135 antibody to human patients in preparation for hematopoietic stem cell transplantation) Using the methods disclosed herein, a physician in the field may provide a hematopoietic stem cell transplant treatment for a patient requiring such treatment. In human patients, antibodies that can bind to antigens expressed by hematopoietic stem cells or Antigen-binding fragments, for example, antibodies that bind to CD135 or their antigen-binding fragments. These can be administered. In this way, the engraftment of hematopoietic stem cell grafts is promoted. This allows the endogenous hematopoietic stem cell population to be depleted before the administration of exogenous hematopoietic stem cell grafts. The antibody is used to control cytotoxicity as described herein or known in the art. It can be covalently conjugated to toxins such as molecules. For example, anti-CD135 antibody or its antigen-binding fragment, Pseudomonas exotoxin A, deBouganin, diphtheria Toxins, amatoxins such as α-amanitin, saporins, maytansine, maytansinoids Auristatin, anthracycline, calicheamicin, irinotecan, SN-38, Ocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indol Nobenzodiazepines, indolinobenzodiazepine dimers, or their variants, etc. It can be conjugated covalently to cytotoxins. This conjugation is Using covalent bond formation techniques described herein or known in the art This can be carried out using an antibody, its antigen-binding fragment, or a drug-antibody condyl. The cetera is then administered to the patient, for example, intravenously, to exogenous hematopoietic stem cells (autologous, homogeneous, Alternatively, it can be administered before transplantation to patients with allogeneic hematopoietic stem cells (or similar).

[0273] The amount of endogenous hematopoietic stem cells is reduced, for example, to 10%, 20%, 30%, 40% before hematopoietic stem cell transplantation. A sufficient amount of anti-CD135 antibody to reduce the population by 50%, 60%, 70%, 80%, 90%, 95%, or more. The antigen-binding fragment or drug-antibody conjugate can be administered. The decrease in hematopoietic stem cell count can be corrected using conventional techniques known in the field, for example, Characteristic hematopoiesis in blood samples taken from patients at various intervals during conditioning therapy. This can be monitored by FACS analysis of cells expressing stem cell surface antigens. For example... In this profession, internists take blood samples from patients at various points during conditioning therapy. The sample can be collected, and FACS analysis is performed to determine the relative concentration of hematopoietic stem cells in the sample. By using antibodies that bind to the ker antigen, we can elucidate the degree of endogenous hematopoietic stem cell reduction. This can be determined. According to some embodiments, the concentration of hematopoietic stem cells is determined by the anti-CD135 Antibodies, their antigen-binding fragments, or drug-antibody conjugates When the minimum value is reached in response to conditioning therapy, the physician will terminate the conditioning therapy. You may begin preparing the patient for hematopoietic stem cell transplantation.

[0274] Anti-CD135 antibody, its antigen-binding fragment, or drug-antibody conjugate, viscosity adjustment Administer to the patient an aqueous solution containing one or more pharmaceutically acceptable excipients, such as a restorative agent. This can be done. The aqueous solution is a technique described herein or known in the art. It can be sterilized using a technique. Before administering hematopoietic stem cell grafts to the patient, antibodies and other substances are used. Antigen-binding fragments, or drug-antibody conjugates, for example, 0.001 mg / kg to 100 It can be administered to patients in doses of mg / kg. Antibodies, their antigen-binding fragments, and This drug-antibody conjugate optimally promotes the engraftment of exogenous hematopoietic stem cells in patients at the optimal time. For example, 1 hour to 1 week after administration of exogenous hematopoietic stem cell grafts (for example, 1 hour, 2 hours, 3 hours) Hours: 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, It can be administered 24 hours, 2, 3, 4, 5, 6, or 7 days prior to the expected date. Cut.

[0275] After the completion of the conditioning therapy, the patient then underwent further conditioning therapy. Injection of exogenous hematopoietic stem cells (e.g., intravenously) from the same internist or from a different internist. Intravenous injection is available. The physician can administer autologous, allogeneic, or allogeneic hematopoietic stem cells. For example, the injection is 1x10 3 ~1x10 9 Hematopoietic stem cells can be administered to patients at a dose of one kilogram. For example, an internist might take a blood sample from a patient and examine hematopoietic stem cells or cells of the hematopoietic lineage (meganuclei). Spheres, platelets, platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglia Cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural cells Measuring the increase in the concentration of grafts (such as Lar cells, T lymphocytes, and B lymphocytes) after administration. This allows us to monitor the engraftment of hematopoietic stem cell grafts. This analysis, for example, For example, it can be performed for 1 hour to 6 months or more after hematopoietic stem cell transplantation treatment. ba, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 o'clock Intervals: 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours Interval, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks Between 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 Weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 (weeks or longer). The concentration of hematopoietic stem cells or hematopoietic lineage cells is appropriate for the period prior to transplantation. Compared to the concentration of the cell type after transplantation (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%) 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, or any of the above. The above findings indicate an increase in anti-CD135 antibodies, their antigen-binding fragments, or drug-antibodies. Treatment with in vivo conjugates successfully promoted the engraftment of transplanted hematopoietic stem cell grafts. This serves as one indicator of that.

[0276] (Example 3: Administration of anti-CD34 antibody to human patients in preparation for hematopoietic stem cell transplantation) Using the methods disclosed herein, a physician in the field may provide a hematopoietic stem cell transplant treatment for a patient requiring such treatment. In human patients, antibodies that can bind to antigens expressed by hematopoietic stem cells or Antigen-binding fragments, such as antibodies that bind to CD34 or their antigen-binding fragments. In this way, the engraftment of hematopoietic stem cells can be promoted. The endogenous hematopoietic stem cell population can be depleted before the administration of exogenous hematopoietic stem cell grafts. The antibody is subjected to cytotoxic components described herein or known in the art. It can be covalently conjugated to toxins such as children. For example, anti-CD34 antibodies or The antigen-binding fragments include Pseudomonas exotoxin A, deBouganin, and diphtheria toxin. amatoxins such as α-amanitin, saporins, maytansine, maytansinoids, Ulistatin, anthracycline, calicheamicin, irinotecan, SN-38, Duo Calmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolino Cells such as benzodiazepines, indolinobenzodiazepine dimers, or their variants. It can be conjugated to toxins via covalent bonds. This conjugation is the original method. Using covalent bond formation techniques described in the document or known in the art This can be done using antibodies, their antigen-binding fragments, or drug-antibody conjugates. The drug is then administered to the patient, for example, intravenously, to exogenous hematopoietic stem cells (autologous, syngeneic, and It can be administered before transplantation to patients with allogeneic hematopoietic stem cells, etc.

[0277] The amount of endogenous hematopoietic stem cells is reduced, for example, to 10%, 20%, 30%, 40% before hematopoietic stem cell transplantation. A sufficient amount of anti-CD34 antibody to reduce by 50%, 60%, 70%, 80%, 90%, 95%, or more. The antigen-binding fragment or drug-antibody conjugate can be administered. The decrease in hematopoietic stem cell count can be corrected using conventional techniques known in the field, for example, Cond Characteristic hematopoietic stem cells in blood samples taken from patients at various intervals during conditioning therapy. This can be monitored by FACS analysis of cells expressing cell surface antigens, for example. Our internists take blood samples from patients at various points during conditioning therapy. It is possible to take samples and perform FACS analysis to determine the relative concentration of hematopoietic stem cells in the sample. By elucidating the extent of endogenous hematopoietic stem cell reduction using antibodies that bind to the Kerr antigen, we can determine the degree of decrease in endogenous hematopoietic stem cells. It can be determined. According to some embodiments, the concentration of hematopoietic stem cells is determined by the anti-CD34 antibody Conditioning with its antigen-binding fragment, or drug-antibody conjugate. When the value reaches its minimum in response to the therapy, the internist will discontinue the conditioning therapy and hematopoiesis. You can begin preparing the patient for stem cell transplantation therapy.

[0278] Anti-CD34 antibodies, their antigen-binding fragments, or drug-antibody conjugates, viscosity adjustment Administer to the patient an aqueous solution containing one or more pharmaceutically acceptable excipients, such as a restorative agent. This can be done. The aqueous solution is a technique described herein or known in the art. It can be sterilized using a technique. Before administering hematopoietic stem cell grafts to the patient, antibodies and other substances are used. Antigen-binding fragments, or drug-antibody conjugates, for example, 0.001 mg / kg to 100 It can be administered to patients in doses of mg / kg. Antibodies, their antigen-binding fragments, and This drug-antibody conjugate optimally promotes the engraftment of exogenous hematopoietic stem cells in patients at the optimal time. For example, 1 hour to 1 week after administration of exogenous hematopoietic stem cell grafts (for example, 1 hour, 2 hours, 3 hours) Hours: 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, It can be administered 24 hours, 2, 3, 4, 5, 6, or 7 days prior to the expected date. Cut.

[0279] After the completion of the conditioning therapy, the patient then underwent further conditioning therapy. Injection of exogenous hematopoietic stem cells (e.g., intravenously) from the same internist or from a different internist. Intravenous injection is available. The physician can administer autologous, allogeneic, or allogeneic hematopoietic stem cells. For example, the injection is 1x10 3 ~1x10 9 Hematopoietic stem cells can be administered to patients at a dose of one kilogram. For example, an internist might take a blood sample from a patient and examine hematopoietic stem cells or cells of the hematopoietic lineage (meganuclei). Spheres, platelets, platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglia Cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural cells Measuring the increase in the concentration of grafts (such as Lar cells, T lymphocytes, and B lymphocytes) after administration. This allows us to monitor the engraftment of hematopoietic stem cell grafts. This analysis, for example, For example, it can be performed for 1 hour to 6 months or more after hematopoietic stem cell transplantation treatment. ba, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 o'clock Intervals: 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours Interval, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks Between 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 Weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 (weeks or longer). The concentration of hematopoietic stem cells or hematopoietic lineage cells is appropriate for the period prior to transplantation. Compared to the concentration of the cell type after transplantation (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%) 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, or any of the above. The above findings indicate an increase in anti-CD34 antibodies, their antigen-binding fragments, or drugs-anti- Treatment with in vivo conjugates successfully promoted the engraftment of transplanted hematopoietic stem cell grafts. This serves as one indicator of that.

[0280] (Example 4: Administration of anti-CD90 antibody to human patients in preparation for hematopoietic stem cell transplantation) Using the methods disclosed herein, a physician in the field may provide a hematopoietic stem cell transplant treatment for a patient requiring such treatment. In human patients, antibodies that can bind to antigens expressed by hematopoietic stem cells or Antigen-binding fragments, such as antibodies that bind to CD90 or their antigen-binding fragments. In this way, the engraftment of hematopoietic stem cells can be promoted. The endogenous hematopoietic stem cell population can be depleted before the administration of exogenous hematopoietic stem cell grafts. The antibody is subjected to cytotoxic components described herein or known in the art. It can be covalently conjugated to toxins such as children. For example, anti-CD90 antibodies or The antigen-binding fragment (such as an anti-GNNK+CD90 antibody or its antigen-binding fragment) Pseudomonas exotoxin A, deBouganin, diphtheria toxin, α-amanitin, etc. Xyn, saporin, meitansine, meitansinoid, auristatin, anthracycline Phosphorus, calicheamycin, irinotecan, SN-38, duocalmycin, pyrrolobenzodiol Azepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, indolino Conjugates cytotoxins such as zodiazepine dimers or their variants by covalent linkage. This conjugation can be made to work as described herein or This can be carried out using covalent bond formation techniques known in the art. Antibodies, The antigen-binding fragment, or drug-antibody conjugate, is then administered to the patient, for example, statically. Intravenous administration allows for the development of exogenous hematopoietic stem cells (such as autologous, syngeneic, or allogeneic hematopoietic stem cells). It can be administered before transplantation to the patient.

[0281] The amount of endogenous hematopoietic stem cells is reduced, for example, to 10%, 20%, 30%, 40% before hematopoietic stem cell transplantation. A sufficient amount of anti-CD90 antibody to reduce by 50%, 60%, 70%, 80%, 90%, 95%, or more. The antigen-binding fragment or drug-antibody conjugate can be administered. The decrease in hematopoietic stem cell count can be corrected using conventional techniques known in the field, for example, Cond Characteristic hematopoietic stem cells in blood samples taken from patients at various intervals during conditioning therapy. This can be monitored by FACS analysis of cells expressing cell surface antigens, for example. Our internists take blood samples from patients at various points during conditioning therapy. It is possible to take samples and perform FACS analysis to determine the relative concentration of hematopoietic stem cells in the sample. By elucidating the extent of endogenous hematopoietic stem cell reduction using antibodies that bind to the Kerr antigen, we can determine the degree of decrease in endogenous hematopoietic stem cells. It can be determined. According to some embodiments, the concentration of hematopoietic stem cells is determined by the anti-CD90 antibody Conditioning with its antigen-binding fragment, or drug-antibody conjugate. When the value reaches its minimum in response to the therapy, the internist will discontinue the conditioning therapy and hematopoiesis. You can begin preparing the patient for stem cell transplantation therapy.

[0282] Anti-CD90 antibodies, their antigen-binding fragments, or drug-antibody conjugates, viscosity adjustment Administer to the patient an aqueous solution containing one or more pharmaceutically acceptable excipients, such as a restorative agent. This can be done. The aqueous solution is a technique described herein or known in the art. It can be sterilized using a technique. Before administering hematopoietic stem cell grafts to the patient, antibodies and other substances are used. Antigen-binding fragments, or drug-antibody conjugates, for example, 0.001 mg / kg to 100 It can be administered to patients in doses of mg / kg. Antibodies, their antigen-binding fragments, and This drug-antibody conjugate optimally promotes the engraftment of exogenous hematopoietic stem cells in patients at the optimal time. For example, 1 hour to 1 week after administration of exogenous hematopoietic stem cell grafts (for example, 1 hour, 2 hours, 3 hours) Hours: 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, It can be administered 24 hours, 2, 3, 4, 5, 6, or 7 days prior to the expected date. Cut.

[0283] After the completion of the conditioning therapy, the patient then underwent further conditioning therapy. Injection of exogenous hematopoietic stem cells (e.g., intravenously) from the same internist or from a different internist. Intravenous injection is available. The physician can administer autologous, allogeneic, or allogeneic hematopoietic stem cells. For example, the injection is 1x10 3 ~1x10 9 Hematopoietic stem cells can be administered to patients at a dose of one kilogram. For example, an internist might take a blood sample from a patient and examine hematopoietic stem cells or cells of the hematopoietic lineage (meganuclei). Spheres, platelets, platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglia Cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural cells Measuring the increase in the concentration of grafts (such as Lar cells, T lymphocytes, and B lymphocytes) after administration. This allows us to monitor the engraftment of hematopoietic stem cell grafts. This analysis, for example, For example, it can be performed for 1 hour to 6 months or more after hematopoietic stem cell transplantation treatment. ba, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 o'clock Intervals: 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours Interval, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks Between 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 Weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 (weeks or longer). The concentration of hematopoietic stem cells or hematopoietic lineage cells is appropriate for the period prior to transplantation. Compared to the concentration of the cell type after transplantation (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%) 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, or any of the above. The above findings indicate an increase in anti-CD90 antibodies, their antigen-binding fragments, or drugs-anti- Treatment with in vivo conjugates successfully promoted the engraftment of transplanted hematopoietic stem cell grafts. This serves as one indicator of that.

[0284] (Example 5: Administration of anti-CD110 antibody to human patients in preparation for hematopoietic stem cell transplantation) Using the methods disclosed herein, a physician in the field may provide a hematopoietic stem cell transplant treatment for a patient requiring such treatment. In human patients, antibodies that can bind to antigens expressed by hematopoietic stem cells or Antigen-binding fragments, for example, antibodies that bind to CD110 or their antigen-binding fragments. These can be administered. In this way, the engraftment of hematopoietic stem cell grafts is promoted. This allows the endogenous hematopoietic stem cell population to be depleted before the administration of exogenous hematopoietic stem cell grafts. The antibody is used to control cytotoxicity as described herein or known in the art. It can be covalently conjugated to toxins such as molecules. For example, anti-CD110 antibody or its antigen-binding fragment, Pseudomonas exotoxin A, deBouganin, diphtheria Toxins, amatoxins such as α-amanitin, saporins, maytansine, maytansinoids Auristatin, anthracycline, calicheamicin, irinotecan, SN-38, Ocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indol Nobenzodiazepines, indolinobenzodiazepine dimers, or their variants, etc. It can be conjugated covalently to cytotoxins. This conjugation is Using covalent bond formation techniques described herein or known in the art This can be carried out using an antibody, its antigen-binding fragment, or a drug-antibody condyl. The cetera is then administered to the patient, for example, intravenously, to exogenous hematopoietic stem cells (autologous, homogeneous, Alternatively, it can be administered before transplantation to patients with allogeneic hematopoietic stem cells (or similar).

[0285] The amount of endogenous hematopoietic stem cells is reduced, for example, to 10%, 20%, 30%, 40% before hematopoietic stem cell transplantation. A sufficient amount of anti-CD110 antibody to reduce the population by 50%, 60%, 70%, 80%, 90%, 95%, or more. The antigen-binding fragment or drug-antibody conjugate can be administered. The decrease in hematopoietic stem cell count can be corrected using conventional techniques known in the field, for example, Characteristic hematopoiesis in blood samples taken from patients at various intervals during conditioning therapy. This can be monitored by FACS analysis of cells expressing stem cell surface antigens. For example... In this profession, internists take blood samples from patients at various points during conditioning therapy. The sample can be collected, and FACS analysis is performed to determine the relative concentration of hematopoietic stem cells in the sample. By using antibodies that bind to the ker antigen, we can elucidate the degree of endogenous hematopoietic stem cell reduction. This can be determined. According to some embodiments, the concentration of hematopoietic stem cells is determined to be anti-CD110 Antibodies, their antigen-binding fragments, or drug-antibody conjugates When the minimum value is reached in response to conditioning therapy, the physician will terminate the conditioning therapy. You may begin preparing the patient for hematopoietic stem cell transplantation.

[0286] Anti-CD110 antibody, its antigen-binding fragment, or drug-antibody conjugate, viscosity adjustment Administer to the patient an aqueous solution containing one or more pharmaceutically acceptable excipients, such as a restorative agent. This can be done. The aqueous solution is a technique described herein or known in the art. It can be sterilized using a technique. Before administering hematopoietic stem cell grafts to the patient, antibodies and other substances are used. Antigen-binding fragments, or drug-antibody conjugates, for example, 0.001 mg / kg to 100 It can be administered to patients in doses of mg / kg. Antibodies, their antigen-binding fragments, and This drug-antibody conjugate optimally promotes the engraftment of exogenous hematopoietic stem cells in patients at the optimal time. For example, 1 hour to 1 week after administration of exogenous hematopoietic stem cell grafts (for example, 1 hour, 2 hours, 3 hours) Hours: 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, It can be administered 24 hours, 2, 3, 4, 5, 6, or 7 days prior to the expected date. Cut.

[0287] After the completion of the conditioning therapy, the patient then underwent further conditioning therapy. Injection of exogenous hematopoietic stem cells (e.g., intravenously) from the same internist or from a different internist. Intravenous injection is available. The physician can administer autologous, allogeneic, or allogeneic hematopoietic stem cells. For example, the injection is 1x10 3 ~1x10 9 Hematopoietic stem cells can be administered to patients at a dose of one kilogram. For example, an internist might take a blood sample from a patient and examine hematopoietic stem cells or cells of the hematopoietic lineage (meganuclei). Spheres, platelets, platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglia Cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural cells Measuring the increase in the concentration of grafts (such as Lar cells, T lymphocytes, and B lymphocytes) after administration. This allows us to monitor the engraftment of hematopoietic stem cell grafts. This analysis, for example, For example, it can be performed for 1 hour to 6 months or more after hematopoietic stem cell transplantation treatment. ba, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 o'clock Intervals: 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours Interval, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks Between 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 Weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 (weeks or longer). The concentration of hematopoietic stem cells or hematopoietic lineage cells is appropriate for the period prior to transplantation. Compared to the concentration of the cell type after transplantation (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%) 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, or any of the above. The above findings indicate an increase in anti-CD110 antibodies, their antigen-binding fragments, or drug-antibodies. Treatment with in vivo conjugates successfully promoted the engraftment of transplanted hematopoietic stem cell grafts. This serves as one indicator of that.

[0288] (Example 6: Generation of antibodies capable of binding to hematopoietic stem cells by phage display) ) Anti-CD45 (e.g., anti-CD45RO), anti-CD45, to be used in conjunction with the compositions and methods described herein. Exemplary methods for the in vitro evolution of CD135, anti-CD34, anti-CD90, or anti-CD110 antibodies are described below. This is a phage display. A phage display library is a CDR of an antibody or an antibody. Similar regions of body-shaped scaffolding (for example, 10 Code array of the BC, CD, and DE loops in the Fn3 domain It can be created by creating a series of mutations or mutations designed within it. The sequences encoding the template antibodies into which these mutations are introduced are, for example, naive human germ cells. These mutations may be systemic sequences. These mutations are standard mutagenesis known in the art. This can be done using technology. Therefore, each mutant sequence removes one or more amino acid mutations. It encodes antibodies corresponding to the template. Retrovirus and phage display vectors The vector can be manipulated using standard vector construction techniques known in the art. Using a compatible protein expression vector along with a P3 phage display vector, antibody multiplication This allows for the creation of phage display vectors for differentiation.

[0289] Mutant DNA provides sequence diversity, and each transformed phage is encoded by its DNA. It shows one mutation in the initial template amino acid sequence, and a vast number of different but structurally related ones. It yields a phage population (library) that exhibits amino acid sequences. It clearly defines the antibody hypervariable region. Due to its structure, the amino acid mutations introduced into the phage display screen are, overall By changing the binding properties of a binding peptide or domain without significantly altering its molecular structure, It is expected.

[0290] In a typical screening, a phage library is used to test for one of the aforementioned antigens or its etiology. It can be bonded by contacting it with a pitope. This facilitates the separation of the binder and non-binder. Therefore, it is convenient to immobilize the target on a solid support. CD45, CD135, CD34, CD90 Phages having a CD110 binding site form a complex with the target on a solid support. However, unbound phages remain in the solution and can be washed away with excess buffer. Next, change the pH of the buffer to an extreme pH (pH 2 or pH 10), and the ionic strength of the buffer. By altering the binding fur, adding a denaturing agent, or by other known means, the binding fur is modified. It can detach the target.

[0291] The recovered phages can then be amplified through infection of bacterial cells, and currently, non-binding The combined antibody is depleted and binds to CD45 (e.g., CD45RO), CD135, CD34, CD90, or CD110. The screening process can be repeated using a new pool of antibodies enriched with antibodies. It is possible. Even if only a few bound phages are recovered, the phages can be used for subsequent screening repetitions. This is sufficient to amplify the selected phage clones in the binding pool after several selections. The gene sequence encoding an antibody or antigen-binding fragment derived from [the substance] is a conventional method The peptide sequence that confers the phage binding affinity to the target is determined by the following: It becomes clear. During the panning process, the sequence diversity of the population is desirable for peptide-binding antibodies. The number of selections decreases with each round until one remains. The sequence contains a small number of associated antibodies or their antigen-binding agents. It can converge to the ragment. The increase in the number of phages recovered in each selection round is due to the library The convergence of the results is an indicator of whether it occurred within the screening process.

[0292] (Example 7: Production of humanized antibodies that bind to hematopoietic stem cell antigens) Non-human antibodies that bind to CD45 (e.g., CD45RO), CD135, CD34, CD90, or CD110 are examples. For example, humanization can be performed by following the procedure below: Consensus human antibody heavy chain and light The chain sequence is known in the relevant art (e.g., "VBASE" human germline sequence data). Base; Kabat et al. Sequences of Proteins of Immunological Interest, Fifth Edit ion, US Department of Health and Human Services, NIH Publication No. 91 -3242, 1991; Tomlinson et al., J. Mol. Biol. 227:776-798, 1992; and Cox et al.. Eur. See J. Immunol. 24:827-836, 1994, each of which disclosures is based on consensus human antimicrobial activity. (Concerning weight chain and light chain sequences, as incorporated herein by reference). Established Using the procedure described, a person skilled in the art can determine the variable domain framework of a consensus antibody sequence. Residues and CDRs can be identified (e.g., by sequence alignment). Humanization To produce antibodies, the heavy and / or light chain variable domains of the consensus human antibody One or more CD-Rs, as described herein, CD45 (e.g., CD45RO), CD135, CD135, CD34, It can be replaced with one or more corresponding CDRs of non-human antibodies that bind to CD90 or CD110. This CDR exchange is a legacy described herein or known in the art. This can be done using gene editing technology.

[0293] An example of a variable domain in a consensus human antibody is the heavy chain variable domain. EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYAMSWVRQAPGKGLEWVAVISENGSDTYYADSVKGRFTISRDDSKNTLY LQMNSLRAEDTAVYYCARDRGGAVSYFDVWGQGTLVTVSS (SEQ ID NO: 34) and Light Chain Variable Domain DI QMTQSPSSLSASVGDRVTITCRASQDVSSYLAWYQQKPGKAPKLLIYAASSLESGVPSRFSGSGSGTDFTLTISSLQPED Includes FATYYCQQYNSLPYTFGQGTKVEIKRT (SEQ ID NO: 35), identified in U.S. Patent No. 6,054,297 The disclosure is made in reference to the Human Antibody Consensus Sequence, and is made in reference to the present invention. It will be incorporated into the detailed specifications. CDR in the above sequence is shown in bold.

[0294] To produce humanized antibodies, one or more variable regions CDRs are CD45 (e.g., CD45RO), CD13 5. A non-human antibody that binds to CD34, CD90, or CD110 is replaced with one or more variable region CDR sequences. By recombinantly expressing the polynucleotide that encodes the above consensus sequence... This is possible. The affinity of antibodies to hematopoietic stem cell antigens is mainly determined by the CDR sequence. Therefore, the obtained humanized antibody has almost the same affinity as the non-human antibody from which the humanized antibody originates. It is expected to show affinity for cellular antigens. This determines the affinity of the antibody against the target antigen. For example, the method described herein and known in the art is an ELISA-based method. The techniques, as well as surface plasmon resonance, fluorescence anisotropy, and isothermal titration calorimetry, are particularly noteworthy. It can be listed.

[0295] (Example 8: Ability of anti-CD45 antibody-drug conjugate to deplete CD45+ cell populations) To investigate the ability of anti-CD45 antibody-drug conjugates to kill CD45+ Reh cells, various studies were conducted. A series of isotype anti-CD45 monoclonal antibodies Ab1, Ab2, and Ab3 are saporincon The jugate Fab fragment was then attached and incubated with CD45+ Reh cells for 3 days. As a negative control, an isotype-matched antibody-saporin conjugate was used as Ab1, Ab Prepare cells 2 and Ab3 respectively, and incubate them with CD45+ Reh cells for 3 days. The cells were then incubated. Cell viability was subsequently evaluated using the CellTiter-Glo® assay. As shown in Figure 1, various antibody-saporin conjugates exhibit different potencies in CD4 It can kill 5+ Reh cells, each dose-dependently depleting CD45+ Reh cell lines. It was done.

[0296] Anti-CD45 monoclonal antibody-saporin conjugate in vitro in human CD34+ hematopoietic stem cells To investigate its ability to deplete the human IgG1 anti-CD45 monoclonal antibody-saporin condyloma, Matched morphogenetic or isotype controls were inked with CD34+ cells at various concentrations. Incubation was performed. After a 6-day incubation period, flow cytometry was used. Cell survival was continuously evaluated. The results of this experiment are reported in Figure 2. As shown there, The anti-CD45 antibody-saporin conjugate dose-dependently depletes human CD34+ hematopoietic stem cells. It was discovered that this is possible.

[0297] In summary, these results were obtained using anti-CD45 antibodies conjugated to saporin. This depletes the population of hematopoietic stem cells in the subject, for example, in preparation for hematopoietic stem cell transplantation therapy. This demonstrates that it is possible to provide a niche where hematopoietic stem cells can home.

[0298] (Other embodiments) All publications, patents, and patent applications referred to herein are as if they were solely the property of their respective owners. It is indicated that the established publication or patent application is incorporated by specific and individual reference. To the same extent, it is incorporated herein by reference.

[0299] Although the present invention has been described in relation to its specific embodiments, it is subject to further modification. It is understood that, generally speaking, this application is based on the principles of the present invention and is known in the art. Any deviation from the present invention that falls within the scope of the invention or customary practice Any modification, use, or adaptation to which the present invention applies in relation to the essential features described above. It is possible to do so, and is intended to be comprehensive to the extent that it conforms to the claims.

[0300] Other embodiments are described in the claims.

Claims

1. A method for depleting a population of CD45RO+ cells in a human patient, wherein the patient is subjected to the treatment of CD45RO The step of administering an effective amount of an antibody or antigen-binding fragment that can bind to it. Includes, method.

2. A method to deplete the CD45RO+ cell population in human patients requiring hematopoietic stem cell transplantation. The patient has an antibody or antigen-binding fragment that can bind to CD45RO. A method comprising the step of administering an effective amount of to before the patient receives a graft containing hematopoietic stem cells. Law.

3. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody or antigen-binding fragment capable of binding to CD45RO is administered to the patient. A method in which a population of CD45RO+ cells is pre-administered in an amount sufficient to deplete them.

4. a. In human patients, an antibody or antigen-binding fragment capable of binding to CD45RO is administered. The steps include administering an amount sufficient to deplete the population of CD45RO+ cells in the patient; b. The next step is to administer a graft containing hematopoietic stem cells to the patient. A method that includes [this].

5. The antibody or its antigen-binding fragment is conjugated to a cytotoxin. The method described in any one of the requests 1 to 4.

6. A method for depleting a population of CD135+ cells in a human patient, wherein the patient is given CD135 The process includes administering an effective amount of an antibody or antigen-binding fragment that can bind to the antibody. Here, the antibody or its antigen-binding fragment is conjugated to a cytotoxin. The method.

7. A method to deplete the CD135+ cell population in human patients requiring hematopoietic stem cell transplantation. Therefore, the patient was given an antibody or antigen-binding fragment capable of binding to CD135. The process includes administering an effective amount of the above to the patient before hematopoietic stem cell grafts are received, And the antibody or its antigen-binding fragment is conjugated to a cytotoxin. method.

8. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody or antigen-binding fragment capable of binding to CD135 is administered to the patient. The population of CD135+ cells is administered in an amount sufficient to deplete it, and here, the anti A method in which the body or its antigen-binding fragments are conjugated to a cytotoxin.

9. a. In human patients, an antibody or antigen-binding fragment capable of binding to CD135 is administered. The steps include administering an amount sufficient to deplete the population of CD135+ cells in the patient; b. The next step is to administer a graft containing hematopoietic stem cells to the patient. This includes, where the antibody or its antigen-binding fragment conjugates the cytotoxin. The method of doing it.

10. The antibody or its antigen-binding fragment includes the following complementarity-determining region (CDR): The method according to any one of claims 6 to 9: a. CDR-H1 having the amino acid sequence SYYMH (SEQ ID NO: 1); b. CDR-H2 having the amino acid sequence IINPSGGSTSYAQKFQG (SEQ ID NO: 2); c. CDR having amino acid sequence GVGAHDAFDI (SEQ ID NO: 3) or VVAAAVADY (SEQ ID NO: 4) -H3； d. Amino acid sequence RSSQSLLHSNGNNYLD (SEQ ID NO: 5) or RSSQSLLHSNGYNYLD (SEQ ID NO: 6) ) CDR-L1; e. CDR-L2 having the amino acid sequence LGSNRAS (SEQ ID NO: 7); and f. CDR- having amino acid sequence MQGTHPAIS (SEQ ID NO: 8) or MQSLQTPFT (SEQ ID NO: 9) L3。

11. The antibody or its antigen-binding fragment comprises one of the following CDRs, according to claims 6 to 9. Methods described in any one of the following items: a. CDR-H1 having the amino acid sequence SYAIS (SEQ ID NO: 10); b. CDR-H2 having the amino acid sequence GIIPIFGTANYAQKFQG (SEQ ID NO: 11); c. CDR-H3 having the amino acid sequence FALFGFREQAFDI (SEQ ID NO: 12); d. CDR-L1 having the amino acid sequence RASQSISSYLN (SEQ ID NO: 13); e. CDR-L2 having the amino acid sequence AASSLQS (SEQ ID NO: 14); and f. CDR-L3 having the amino acid sequence QQSYSTPFT (SEQ ID NO: 15).

12. A method for depleting a population of CD135+ cells in a human patient, wherein the patient is given CD135 The process involves administering an effective amount of a human Flt3 ligand or its fragment that can bind to it. Methods that include the degree.

13. A method to deplete the CD135+ cell population in human patients requiring hematopoietic stem cell transplantation. Therefore, to the aforementioned patient, a human Flt3 ligand capable of binding to CD135, or its flag The process includes administering an effective amount of ment to the patient before hematopoietic stem cell grafts are received. ,method.

14. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient A human Flt3 ligand or fragment thereof that can bind to CD135, the patient A method in which a sufficient amount is pre-administered to deplete the population of CD135+ cells in the subject.

15. a. Human patients with a human Flt3 ligand capable of binding to CD135, or its fragmentation The process involves administering the drug in an amount sufficient to deplete the population of CD135+ cells in the patient. ； b. The next step is to administer a graft containing hematopoietic stem cells to the patient. A method that includes [this].

16. The human Flt3 ligand or its fragment is covalently bound to the Fc domain. The method described in any one of the requests 12 to 15.

17. The N-terminus of the human Flt3 ligand or its fragment is covalently bound to the Fc domain. The method according to claim 16.

18. The C-terminus of the human Flt3 ligand or its fragment is covalently bound to the Fc domain. The method according to claim 16.

19. The Fc domain is covalently bound to the cytotoxin, according to any one of claims 16 to 18. Method of description.

20. The claim is that the human Flt3 ligand or a fragment thereof is covalently bound to the cytotoxin. The method described in any one of items 12 to 15.

21. The N-terminus of the human Flt3 ligand or its fragment is covalently bound to the cytotoxin. The method according to claim 20.

22. The C-terminus of the human Flt3 ligand or its fragment is covalently bound to the cytotoxin. The method according to claim 20.

23. The cytotoxin is covalently bound to the Fc domain, according to any one of claims 20 to 22. Method of description.

24. The human Flt3 ligand or its fragment is covalently bonded to the cytotoxin at one site. The method according to claim 20, wherein the Fc domain is covalently bonded at different sites.

25. The Fc domain is a human IgG1, IgG2, IgG3, or IgG4 isotype Fc domain. The method according to any one of claims 16 to 19, 23, and 24.

26. A method for depleting a population of CD34+ cells in a human patient, wherein the patient is linked to CD34 The step includes administering an effective amount of an antibody or antigen-binding fragment that can be combined. Here, the antibody or its antigen-binding fragment is conjugated to a cytotoxin. There is a way.

27. A method to deplete the CD34+ cell population in human patients requiring hematopoietic stem cell transplantation. Therefore, the patient was given an antibody or antigen-binding fragment that can bind to CD34. The step includes administering an effective amount to the patient before hematopoietic stem cell grafts are received, wherein The antibody or its antigen-binding fragment is conjugated to a cytotoxin. Law.

28. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody or antigen-binding fragment capable of binding to CD34 is used in the patient. The antibody is administered in an amount sufficient to deplete the population of CD34+ cells, and here, Alternatively, a method in which the antigen-binding fragment is conjugated to a cytotoxin.

29. a. In human patients, an antibody or antigen-binding fragment capable of binding to CD34 is administered. The steps include administering an amount sufficient to deplete the population of CD34+ cells in the patient; b. The next step is to administer a graft containing hematopoietic stem cells to the patient. This includes, where the antibody or its antigen-binding fragment conjugates the cytotoxin. The method of doing it.

30. A method for depleting a population of CD90+ cells in a human patient, wherein the patient is linked to CD90 The step includes administering an effective amount of an antibody or antigen-binding fragment that can be combined. Here, the antibody or its antigen-binding fragment is conjugated to a cytotoxin. There is a way.

31. A method to deplete the CD90+ cell population in human patients requiring hematopoietic stem cell transplantation. Therefore, the patient was given an antibody or antigen-binding fragment that can bind to CD90. The step includes administering an effective amount to the patient before hematopoietic stem cell grafts are received, wherein The antibody or its antigen-binding fragment is conjugated to a cytotoxin. Law.

32. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody or antigen-binding fragment capable of binding to CD90 is used in the patient. The antibody is administered in an amount sufficient to deplete the population of CD90+ cells, and here, Alternatively, a method in which the antigen-binding fragment is conjugated to a cytotoxin.

33. a. In human patients, an antibody or antigen-binding fragment capable of binding to CD90 is administered. The steps include administering an amount sufficient to deplete the population of CD90+ cells in the patient; b. The next step is to administer a graft containing hematopoietic stem cells to the patient. This includes, where the antibody or its antigen-binding fragment conjugates the cytotoxin. The method of doing it.

34. A method for depleting a population of CD110+ cells in a human patient, wherein the patient has CD110 The process includes administering an effective amount of an antibody or antigen-binding fragment that can bind to the antibody. Here, the antibody or its antigen-binding fragment is conjugated to a cytotoxin. The method.

35. A method to deplete the population of CD110+ cells in human patients requiring hematopoietic stem cell transplantation. Therefore, the patient was given an antibody or antigen-binding fragment capable of binding to CD110. The process includes administering an effective amount of the above to the patient before hematopoietic stem cell grafts are received, And the antibody or its antigen-binding fragment is conjugated to a cytotoxin. method.

36. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody or antigen-binding fragment capable of binding to CD110 is administered to the patient. The drug is administered in an amount sufficient to deplete the population of CD110+ cells, and here, the anti A method in which the body or its antigen-binding fragments are conjugated to a cytotoxin.

37. a. In human patients, an antibody or antigen-binding fragment capable of binding to CD110 is administered. The steps include administering an amount sufficient to deplete the population of CD110+ cells in the patient; b. The next step is to administer a graft containing hematopoietic stem cells to the patient. This includes, where the antibody or its antigen-binding fragment conjugates the cytotoxin. The method of doing it.

38. The aforementioned cytotoxins include amatoxin, Pseudomonas exotoxin A, deBouganin, and diphtheria toxin. Substances, saporins, meitansine, meitansinoids, auristatin, anthracyclines , calicheamycin, irinotecan, SN-38, duocalmycin, pyrrolobenzodiaze Pyrrolobenzodiazepine dimer, indolinobenzodiazepine, and indolino A selected from the group consisting of benzodiazepine dimers or their variants, claims 5 to The method described in any one of items 11 and 19-37.

39. A method for depleting a population of CD45+ cells in a human patient, wherein the patient is linked to CD45 The step includes administering an effective amount of an antibody or antigen-binding fragment that can be combined. Here, the antibody or its antigen-binding fragment is amatoxin, Pseudomonas Exotoxin A, deBouganin, diphtheria toxin, saporin, meitansine, meitansinoid, Auristatin, anthracycline, calicheamicin, irinotecan, SN-38, du Ocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolin Benzodiazepines, and indolinobenzodiazepine dimers, or their variants A method that conjugates cytotoxins selected from a group of substances.

40. A method to deplete the CD45+ cell population in human patients requiring hematopoietic stem cell transplantation. Therefore, the patient was given an antibody or antigen-binding fragment that can bind to CD45. The step includes administering an effective amount to the patient before hematopoietic stem cell grafts are received, wherein The antibody or its antigen-binding fragment is amatoxin, Pseudomonas exotoxin A deBouganin, diphtheria toxin, saporin, maytansine, maytansinoid, auris Tatin, anthracycline, calicheamicin, irinotecan, SN-38, Duokarma Icin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine A group consisting of azepines and indolinobenzodiazepine dimers, or their variants. A method that conjugates to more selective cytotoxins.

41. A method comprising the step of administering a graft containing hematopoietic stem cells to a human patient, wherein the patient an antibody or antigen-binding fragment capable of binding to CD45 is used in the patient. The antibody is administered in an amount sufficient to deplete the population of CD45+ cells, and here, or its antigen-binding fragment is amatoxin, Pseudomonas exotoxin A, deBougani n, diphtheria toxin, saporin, maytansine, maytansinoid, auristatin, a Intracycline, calicheamycin, irinotecan, SN-38, duocalmycin, pi Lolobenzodiazepine, pyrrolobenzodiazepine dimer, indolinobenzodiazepine, and selected from the group consisting of indolinobenzodiazepine dimers or their variants. A method that involves conjugating the cytotoxins being produced.

42. a. In human patients, an antibody or antigen-binding fragment capable of binding to CD45 is administered. The steps include administering an amount sufficient to deplete the population of CD45+ cells in the patient; b. The next step is to administer a graft containing hematopoietic stem cells to the patient. This includes, where the antibody or its antigen-binding fragment is amatoxin, shu Domonas exotoxin A, deBouganin, diphtheria toxin, saporin, meitansine, meitansine Noid, auristatin, anthracycline, calicheamicin, irinotecan, SN-3 8. Duocalmycin, pyrrolobenzodiazepine, pyrrolobenzodiazepine dimer, i Indolinobenzodiazepines, and indolinobenzodiazepine dimers, or their A method that involves conjugating a cytotoxin selected from a group of mutants.

43. The antibody conjugated to a cytotoxin, its antigen-binding fragment, or ligand However, it is represented by the formula Ab-Am, where Ab is the antibody, its antigen-binding fragment, or liggan. Claims 5-11 and 19-4, wherein Am is an amatoxin represented by formula (I). The method described in any one of paragraphs 2: 【Chemistry 1】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 is H, R C or R D and; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 Heteroalkylene, C replaced with intention 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, arbitrarily Substituted C 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, optionally substituted substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted Arrinenes, or optionally substituted heteroarrinenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

44. The method according to claim 43, wherein Am is an amatoxin represented by formula (IA). 【Chemistry 2】

45. The method according to claim 43, wherein Am is an amatoxin represented by formula (IB). 【Transformation 3】

46. R A and R B However, together with the oxygen atoms to which they are bonded, they combine to form the following: The method according to claim 44 or 45: 【Chemistry 4】 [In the formula, Y is O, S, NR E , and CR E R E Selected from, and R E and R E' Each of these is independently and arbitrarily substituted C 1 -C 6 Alkilen-R C , replace as desired C 1 -C 6 Heteroalkylene-R C , arbitrarily substituted C 2 -C 6 Alkenylene-R C , replace as desired C 2 -C 6 Heteroalkenylene-R C , arbitrarily substituted C 2 -C 6 Alkinylene-R C , place as you like Replaced C 2 -C 6 Heteroalkynylene-R C , optionally substituted cycloalkylene-R C , at will Substituted heterocycloalkylene-R C , optionally substituted arylene-R C , or optionally Substituted heteroarylene-R C It is.

47. R A and R B However, together with the oxygen atoms to which they are bonded, they combine to form the following: The method according to claim 46: 【Transformation 5】 。

48. R 1 is H, OH, or OR A ; R 2 However, H, OH, or OR B And; R A and R B However, together with the oxygen atoms to which they are bonded, they combine to form the following: ： 【Transformation 6】 ； R 3 、R 4 、R 6 、and R 7 are each H; R 5 is OR C And; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 44 or 45, wherein is H or OH.

49. R 1 and R 2 However, each is independently either H or OH; R 3 R C And; R 4 , R 6 , and R 7 However, each of them is H; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 44 or 45, wherein is H or OH.

50. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 and R 5 However, each is independent of H, OH, and OR C , or R C And; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 44 or 45, wherein is H or OH.

51. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 and R 5 However, each is independently either H or OH; R 8 However, OR C or NHR C and; as well R 9 The method according to claim 44 or 45, wherein is H or OH.

52. The antibody conjugated to a cytotoxin, its antigen-binding fragment, or ligand However, it is represented by the formula Ab-Am, where Ab is the antibody, its antigen-binding fragment, or liggan. Claims 5-11 and 19-4, wherein Am is an amatoxin represented by formula (II). The method described in any one of paragraphs 2: 【Transformation 7】 [In the formula, X is S, SO, or SO 2 And; R 1 However, phosphorus covalently bound to H, or the antibody or its antigen-binding fragment. Car; and R 2 However, phosphorus covalently bound to H, or the antibody or its antigen-binding fragment. It is a car; Here, R 1 If H, then R 2 The linker is R 2 If H, then R 1 is the phosphorus It is a car.

53. The cytotoxin is a maytansinoid selected from the group consisting of DM1 and DM4. The method according to any one of the requests 5 to 11 and 19 to 42.

54. The cytotoxin consists of monomethyl auristatin E and monomethyl auristatin F. Auristatin selected from the group, any one of claims 5 to 11 and 19 to 42. The method described in item 1.

55. The cytotoxins mentioned above are daunorubicin, doxorubicin, epirubicin, and idarubicin. An anthracycline selected from the group consisting of n, claims 5-11 and 19-4 The method described in any one of item 2.

56. The antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding Fragments, polyclonal antibodies or their antigen-binding fragments, humanized antibodies or The antigen-binding fragment, the bispecific antibody or its antigen-binding fragment, bivariable Immunoglobulin domain, single-stranded Fv molecule (scFv), diabody, triabody, nanobody D, antibody-like protein scaffold, Fv fragment, Fab fragment, F(ab') 2 molecules, and Any one of claims 1 to 11 and 26 to 55, selected from the group consisting of tandem scFV. The method described in item 1.

57. The antibody is an isotype selected from the group consisting of IgG, IgA, IgM, IgD, and IgE. The method according to any one of claims 1 to 11 and 26 to 56, comprising:

58. The antibody, its antigen-binding fragment, or ligand, after administration to the patient, Any one of claims 1 to 57, which is taken up by rhinocytes, autoimmune cells, or hematopoietic stem cells. The method described in section [section number].

59. The aforementioned antibody, its antigen-binding fragment, or ligand is used by cancer cells, autoimmune cells, Or, according to any one of claims 1 to 58, which can promote necrosis of hematopoietic stem cells. The method.

60. When the antibody, its antigen-binding fragment, or ligand is administered to the patient, Claims 2 to 5, 7, which allow one or more complement proteins to be supplied to the hematopoietic stem cells. Any of the following: ~11, 13~25, 27~29, 31~33, 35~38, and 40~59 The method described in item 1.

61. The graft containing hematopoietic stem cells contains the antibody, its antigen-binding fragment, or ligan Claims 2-5, the drug is administered to the patient after its concentration has been substantially removed from the patient's blood. , 7-11, 13-25, 27-29, 31-33, 35-38, and 40-60 The method described in either of the above terms.

62. The hematopoietic stem cells or their offspring have been in the patient for at least two days since the transplantation of the hematopoietic stem cells into the patient. Later, claims 2-5, 7-11, 13-25, which maintain the functional capacity of hematopoietic stem cells, The method according to any one of items 27-29, 31-33, 35-38, and 40-61.

63. The hematopoietic stem cells or their offspring, after transplantation of the hematopoietic stem cells into the patient, Claims 2-5, 7- 11, 13-25, 27-29, 31-33, 35-38, and 40-62 The method described in item 1.

64. During transplantation to the aforementioned patient, the hematopoietic stem cells are megakaryocytes, platelets, platelets, red blood cells, and obesity cells. Cells, myeloblasts, basophils, neutrophils, eosinophils, microglial cells, granulocytes, monocytes, osteoclasts, antigens Presenting cells, macrophages, dendritic cells, natural killer cells, T lymphocytes, and B lymphocytes Claims 2-5, 7-11, which cause the recovery of a cell population selected from a group consisting of pocytes. In any one of the following items: 13-25, 27-29, 31-33, 35-38, and 40-63 Method of description.

65. The method according to any one of claims 1 to 64, wherein the patient is suffering from stem cell dysfunction.

66. The patient is suffering from an abnormal hemoglobin disorder, according to any one of claims 1 to 65. Method of description.

67. The aforementioned abnormal hemoglobin disorders include sickle cell anemia, thalassemia, Fanconi anemia, and regeneration. A claim is made from a group consisting of atypical anemia and Wiscott-Aldrich syndrome. The method described in paragraph 66.

68. The method according to claim 66, wherein the abnormal hemoglobin disorder is Fanconi anemia.

69. The method according to claim 66, wherein the abnormal hemoglobin disorder is aplastic anemia.

70. The method according to claim 66, wherein the abnormal hemoglobin disorder is sickle cell anemia.

71. The method according to claim 66, wherein the abnormal hemoglobin disorder is thalassemia.

72. The person according to any one of claims 1 to 71, wherein the patient is suffering from myelodysplastic disorder. Law.

73. The method according to any one of claims 1 to 72, wherein the patient suffers from an immunodeficiency disorder. 。

74. The method according to claim 73, wherein the immunodeficiency disorder is congenital immunodeficiency.

75. The method according to claim 73, wherein the immunodeficiency disorder is acquired immunodeficiency.

76. If the aforementioned acquired immunodeficiency is human immunodeficiency virus or acquired immunodeficiency syndrome, please The method described in item 75.

77. The method according to any one of claims 1 to 76, wherein the patient is suffering from a metabolic disorder.

78. The aforementioned metabolic disorders include glycogen storage disease, mucopolysaccharidosis, Gaucher disease, Haller's disease, and sphygmomanitis. A claim selected from the group consisting of ngolipidosis and metachromatic leukodystrophy. Method 77.

79. The method according to any one of claims 1 to 78, wherein the patient is suffering from cancer.

80. The aforementioned cancers are selected from the group consisting of leukemia, lymphoma, multiple myeloma, and neuroblastoma. The method according to claim 79, which is selected.

81. The method according to claim 79, wherein the cancer is a blood cancer.

82. The method according to claim 79, wherein the cancer is acute myeloid leukemia.

83. The method according to claim 79, wherein the cancer is acute lymphoblastic leukemia.

84. The method according to claim 79, wherein the cancer is chronic myeloid leukemia.

85. The method according to claim 79, wherein the cancer is chronic lymphocytic leukemia.

86. The method according to claim 79, wherein the cancer is diffuse large B-cell lymphoma.

87. The method according to claim 79, wherein the cancer is non-Hodgkin lymphoma.

88. The aforementioned patient has adenosine deaminase deficiency and severe combined immunodeficiency, hyperimmune globulinia. M syndrome, Chediak-Higashi syndrome, hereditary lymphohistiocytosis, osteopetrosis, bone formation Thalassemia dysplasia, thalassemia major, systemic sclerosis, systemic lupus erythematosus, multiple If you have a disorder selected from the group consisting of juvenile sclerosis and juvenile rheumatoid arthritis, please The method described in any one of the requests 1 to 87.

89. The method according to any one of claims 1 to 88, wherein the patient suffers from an autoimmune disease. 。

90. The method according to claim 89, wherein the autoimmune disorder is selected from the group consisting of the following: multiple Treatment for: sclerosing, human systemic lupus, rheumatoid arthritis, inflammatory bowel disease, psoriasis, type 1 diabetes mellitus. Acute disseminated encephalomyelitis, Addison's disease, alopecia generalis, ankylosing spondylitis, antiphospholipid antibody syndrome group, aplastic anemia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune otitis interna, autoimmune Immunoproliferative syndrome, autoimmune oophoritis, Barlow's disease, Behçet's disease, bullous celestial artery Smallpox, cardiomyopathy, Chagas disease, chronic fatigue immunodeficiency syndrome, chronic inflammatory demyelinating polyneuropathy, Rohn's disease, cicatricial pemphigoid, celiac disease-dermatitis herpetiformis, cold agglutinin disease, CREST syndrome, Dogo's disease, lupus discoid, autonomic nervous system dysfunction, endometriosis, essential mixed cryoglobulinemia Fibromyalgia-fibromyositis, Goodpasture syndrome, Graves' disease, Guillain-Barré syndrome Group, Hashimoto's thyroiditis, hidradenitis suppurativa, idiopathic and / or acute thrombocytopenic purpura, idiopathic Pulmonary fibrosis, IgA neuropathy, interstitial cystitis, juvenile arthritis, Kawasaki disease, lichen planus, Lyme disease Meniere's disease, mixed connective tissue disease, myasthenia gravis, neurogenic tonicity, opsoclonus Myoclonus syndrome, optic neuritis, oat thyroiditis, pemphigus vulgaris, pernicious anemia, multiple molluscum contagiosum Osteitis, polymyositis and dermatomyositis, primary biliary cirrhosis, polyarteritis nodosa, polyglandular syndrome Polymyalgia rheumatica, primary agammaglobulinemia, Raynaud's phenomenon, Reiter's syndrome Rheumatic fever, sarcoidosis, scleroderma, Sjögren's syndrome, Stiff Person syndrome Group, Takayasu's arteritis, temporal arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, vulvodynia, Wegener's granulomatosis.

91. The method according to claim 89, wherein the autoimmune disorder is scleroderma.

92. The method according to claim 89, wherein the autoimmune disorder is multiple sclerosis.

93. The method according to claim 89, wherein the autoimmune disorder is ulcerative colitis.

94. The method according to claim 89, wherein the autoimmune disorder is Crohn's disease.

95. The method according to claim 89, wherein the autoimmune disorder is type 1 diabetes.

96. The method described in any one of claims 65 to 95, wherein the method treats the disorder or cancer. method.

97. A method for depleting a population of CD45+ cells, wherein the population is a conjugate represented by the formula Ab-Am. The process includes a step of contacting an effective amount of the agent, wherein Ab is an antibody or antigen that binds to CD45. A binding fragment, and Am is an amatoxin, in this method.

98. The method according to claim 97, wherein Am is represented by formula (IA): 【Transformation 8】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 heteroalkylene, Arbitrarily substituted C 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, optional C replaced 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Arylenes, or optionally substituted heteroarylenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

99. The method according to claim 97, wherein Am is represented by formula (IB): 【Chemistry 9】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 heteroalkylene, Arbitrarily substituted C 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, optional C replaced 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Arylenes, or optionally substituted heteroarylenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

100. The antibody or its antigen-binding fragment It is conjugated to the amatoxin via cysteine ​​residues in the Fc domain of the . The method according to claim 97.

101. The cysteine ​​residue is in the Fc domain of the antibody or its antigen-binding fragment. The method according to claim 100, which is introduced by a mutation.

102. The cysteine ​​residue is selected from the group consisting of Cys118, Cys239, and Cys265. The method described in item 101.

103. The cysteine ​​residue is naturally present in the Fc domain of the antibody or its antigen-binding fragment. The method according to claim 100.

104. The Fc domain is an IgG Fc domain, and the cysteine ​​residues are Cys261, Cys321, and Cys3 The method according to claim 103, selected from the group consisting of 67 and Cys425.

105. R 1 However, H, OH, or OR A And; R 2 However, H, OH, or OR B And; R A and R B However, together with the oxygen atoms to which they are bonded, they combine to form the following: ： 【Chemistry 10】 ； R 3 , R 4 , R 6 , and R 7 However, each of them is H; R 5 is OR C And; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 98 or 99, wherein is H or OH.

106. R 1 and R 2 However, each is independently either H or OH; R 3 R C And; R 4 , R 6 , and R 7 However, each of them is H; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 98 or 99, wherein is H or OH.

107. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 However, OR C , or R C And; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 98 or 99, wherein is H or OH.

108. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 and R 5 However, each is independently either H or OH; R 8 However, OR C or NHR C and; as well R 9 The method according to claim 98 or 99, wherein is H or OH.

109. The antibody or its antigen-binding fragment is taken up by CD45+ cells, The method described in item 97.

110. The antibody or its antigen-binding fragment contains approximately 0.1 pM to approximately 1 μM of K d Combine with CD45 The method according to claim 97.

111. The antibody or its antigen-binding fragment is approximately 9 × 10 -2 M -1 s -1 ~Approx. 1×10 2 M -1 s -1 k on The method according to claim 97, wherein the CD45 is coupled to the CD45.

112. The antibody or its antigen-binding fragment is a second antibody or its antigen-binding fragment of CD45. Competitively inhibits binding to the fragment of the second antibody or its antigen-binding fragment. However, the method according to claim 97, comprising the following complementarity determination region (CDR): a. CDR-H1 having the amino acid sequence SYAMS (SEQ ID NO: 16); b. CDR-H2 having the amino acid sequence AISGSGGSTFYADSVRG (SEQ ID NO: 17); c. CDR-H3 having the amino acid sequence EVMGPIFFDY (SEQ ID NO: 18); d. CDR-L1 having the amino acid sequence RASQSIISSALA (SEQ ID NO: 19); e. CDR-L2 having the amino acid sequence GASSRAT (SEQ ID NO: 20); and f. CDR-L3 having the amino acid sequence QQYGSTPLT (SEQ ID NO: 21).

113. The antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding Fragments, polyclonal antibodies or their antigen-binding fragments, humanized antibodies or The antigen-binding fragment, the bispecific antibody or its antigen-binding fragment, bivariable Immunoglobulin domain, single-stranded Fv molecule (scFv), diabody, triabody, nanobody D, antibody-like protein scaffold, Fv fragment, Fab fragment, F(ab') 2 molecules, and The method according to claim 97, selected from the group consisting of tandem scFV.

114. A conjugate represented by the formula Ab-Am, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, and Am is the amatoxin, a conjugate.

115. Am is the conjugate according to claim 114, represented by formula (IA): 【Chemistry 11】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 heteroalkylene, Arbitrarily substituted C 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, optional C replaced 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Arylenes, or optionally substituted heteroarylenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

116. Am is the conjugate according to claim 114, represented by formula (IB): 【Chemistry 12】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 heteroalkylene, Arbitrarily substituted C 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, optional C replaced 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Arylenes, or optionally substituted heteroarylenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

117. The antibody or its antigen-binding fragment It is conjugated to the amatoxin via cysteine ​​residues in the Fc domain of the . , the conjugate according to claim 114.

118. The cysteine ​​residue is in the Fc domain of the antibody or its antigen-binding fragment. The conjugate according to claim 117, which is introduced by a mutation.

119. The cysteine ​​residue is selected from the group consisting of Cys118, Cys239, and Cys265. The conjugate described in item 118.

120. The cysteine ​​residue is naturally present in the Fc domain of the antibody or its antigen-binding fragment. The conjugate described in claim 117, which exists in the present.

121. The Fc domain is an IgG Fc domain, and the cysteine ​​residues are Cys261, Cys321, and Cys3 The conjugate according to claim 120, selected from the group consisting of 67 and Cys425.

122. R 1 However, H, OH, or OR A And; R 2 However, H, OH, or OR B And; R A and R B However, together with the oxygen atoms to which they are bonded, they combine to form the following: ： 【Chemistry 13】 ； R 3 , R 4 , R 6 , and R 7 However, each of them is H; R 5 is OR C And; R 8 However, OH or NH 2 and; as well R 9 The conjugate according to claim 115 or 116, wherein the conjugate is H or OH.

123. R 1 and R 2 However, each is independently either H or OH; R 3 R C And; R 4 , R 6 , and R 7 However, each of them is H; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The conjugate according to claim 115 or 116, wherein the conjugate is H or OH.

124. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 However, OR C , or R C And; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The conjugate according to claim 115 or 116, wherein the conjugate is H or OH.

125. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 and R 5 However, each is independently either H or OH; R 8 However, OR C or NHR C and; as well R 9 The conjugate according to claim 115 or 116, wherein the conjugate is H or OH.

126. The antibody or its antigen-binding fragment is taken up by CD45+ cells, The conjugate described in item 114.

127. The antibody or its antigen-binding fragment contains approximately 0.1 pM to approximately 1 μM of K d Combine with CD45 , the conjugate according to claim 114.

128. The antibody or its antigen-binding fragment is approximately 9 × 10 -2 M -1 s -1 ~Approx. 1×10 2 M -1 s -1 k on The conjugate according to claim 114, which is coupled to CD45.

129. The antibody or its antigen-binding fragment is a second antibody or its antigen-binding fragment of CD45. Competitively inhibits binding to the fragment of the second antibody or its antigen-binding fragment. However, the conjugate according to claim 114 includes the following complementarity determination region (CDR): a. CDR-H1 having the amino acid sequence SYAMS (SEQ ID NO: 16); b. CDR-H2 having the amino acid sequence AISGSGGSTFYADSVRG (SEQ ID NO: 17); c. CDR-H3 having the amino acid sequence EVMGPIFFDY (SEQ ID NO: 18); d. CDR-L1 having the amino acid sequence RASQSIISSALA (SEQ ID NO: 19); e. CDR-L2 having the amino acid sequence GASSRAT (SEQ ID NO: 20); and f. CDR-L3 having the amino acid sequence QQYGSTPLT (SEQ ID NO: 21).

130. The antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding Fragments, polyclonal antibodies or their antigen-binding fragments, humanized antibodies or The antigen-binding fragment, the bispecific antibody or its antigen-binding fragment, bivariable Immunoglobulin domain, single-stranded Fv molecule (scFv), diabody, triabody, nanobody D, antibody-like protein scaffold, Fv fragment, Fab fragment, F(ab') 2 molecules, and The conjugate according to claim 114, selected from the group consisting of tandem scFV.

131. A method for depleting a population of CD135+ cells, wherein the population is represented by the conjugate formula Ab-Am. The process includes contacting an effective amount of the gate, wherein Ab is an antibody that binds to CD135 or A method in which Am is an antigen-binding fragment and is an amatoxin.

132. The method according to claim 131, wherein Am is represented by formula (IA): 【Chemistry 14】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 heteroalkylene, Arbitrarily substituted C 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, optional C replaced 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Arylenes, or optionally substituted heteroarylenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

133. The method according to claim 131, wherein Am is represented by formula (IB): 【Chemistry 15】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 heteroalkylene, Arbitrarily substituted C 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, optional C replaced 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Arylenes, or optionally substituted heteroarylenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

134. The antibody or its antigen-binding fragment It is conjugated to the amatoxin via cysteine ​​residues in the Fc domain of the . The method according to claim 131.

135. The cysteine ​​residue is in the Fc domain of the antibody or its antigen-binding fragment. The method according to claim 134, which is introduced by a mutation.

136. The cysteine ​​residue is selected from the group consisting of Cys118, Cys239, and Cys265. The method described in item 135.

137. The cysteine ​​residue is naturally present in the Fc domain of the antibody or its antigen-binding fragment. The method according to claim 134.

138. The Fc domain is an IgG Fc domain, and the cysteine ​​residues are Cys261, Cys321, and Cys3 The method according to claim 137, selected from the group consisting of 67 and Cys425.

139. R 1 However, H, OH, or OR A And; R 2 However, H, OH, or OR B And; R A and R B However, together with the oxygen atoms to which they are bonded, they combine to form the following: ： 【Chemistry 16】 ； R 3 , R 4 , R 6 , and R 7 However, each of them is H; R 5 is OR C And; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 132 or 133, wherein the compound is H or OH.

140. R 1 and R 2 However, each is independently either H or OH; R 3 R C And; R 4 , R 6 , and R 7 However, each of them is H; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 132 or 133, wherein the compound is H or OH.

141. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 However, OR C , or R C And; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The method according to claim 132 or 133, wherein the compound is H or OH.

142. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 and R 5 However, each is independently either H or OH; R 8 However, OR C or NHR C and; as well R 9 The method according to claim 132 or 133, wherein the compound is H or OH.

143. The antibody or its antigen-binding fragment is taken up by CD135+ cells, The method described in item 131.

144. The antibody or its antigen-binding fragment contains approximately 0.1 pM to approximately 1 μM of K d Combine with CD135 The method according to claim 131.

145. The antibody or its antigen-binding fragment is approximately 9 × 10 -2 M -1 s -1 ~Approx. 1×10 2 M -1 s -1 k on The method according to claim 131, wherein the CD135 is coupled to the method according to claim 131.

146. The antibody or its antigen-binding fragment is a second antibody or its antigen-binding fragment of CD135. Competitively inhibits binding to the fragment of the second antibody or its antigen-binding fragment. However, the method according to claim 131, comprising the following complementarity determination region (CDR): a. CDR-H1 having the amino acid sequence SYYMH (SEQ ID NO: 1); b. CDR-H2 having the amino acid sequence IINPSGGSTSYAQKFQG (SEQ ID NO: 2); c. CDR having amino acid sequence GVGAHDAFDI (SEQ ID NO: 3) or VVAAAVADY (SEQ ID NO: 4) -H3； d. Amino acid sequence RSSQSLLHSNGNNYLD (SEQ ID NO: 5) or RSSQSLLHSNGYNYLD (SEQ ID NO: 6) ) CDR-L1; e. CDR-L2 having the amino acid sequence LGSNRAS (SEQ ID NO: 7); and f. CDR- having amino acid sequence MQGTHPAIS (SEQ ID NO: 8) or MQSLQTPFT (SEQ ID NO: 9) L3。

147. The antibody or its antigen-binding fragment is a second antibody or its antigen-binding fragment of CD135. Competitively inhibits binding to the fragment of the second antibody or its antigen-binding fragment. However, the method according to claim 131, comprising the following complementarity determination region (CDR): a. CDR-H1 having the amino acid sequence SYAIS (SEQ ID NO: 10); b. CDR-H2 having the amino acid sequence GIIPIFGTANYAQKFQG (SEQ ID NO: 11); c. CDR-H3 having the amino acid sequence FALFGFREQAFDI (SEQ ID NO: 12); d. CDR-L1 having the amino acid sequence RASQSISSYLN (SEQ ID NO: 13); e. CDR-L2 having the amino acid sequence AASSLQS (SEQ ID NO: 14); and f. CDR-L3 having the amino acid sequence QQSYSTPFT (SEQ ID NO: 15).

148. The antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding Fragments, polyclonal antibodies or their antigen-binding fragments, humanized antibodies or The antigen-binding fragment, the bispecific antibody or its antigen-binding fragment, bivariable Immunoglobulin domain, single-stranded Fv molecule (scFv), diabody, triabody, nanobody D, antibody-like protein scaffold, Fv fragment, Fab fragment, F(ab') 2 molecules, and The method according to claim 131, selected from the group consisting of tandem scFV.

149. A conjugate represented by the formula Ab-Am, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, and Am is the amatoxin, a conjugate.

150. Am is the conjugate according to claim 149, represented by formula (IA): 【Chemistry 17】 [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 Heteroalkylene, C replaced with intention 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, arbitrarily Substituted C 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, optionally substituted substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted Arrinenes, or optionally substituted heteroarrinenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

151. Am is the conjugate according to claim 149, represented by formula (IB): [Chemistry 18] [In the formula, R 1 H, OH, OR A , or OR C And; R 2 H, OH, OR B , or OR C And; R A and R B They, together with the oxygen atoms to which they are bonded, bond and optionally substitute. It forms a 5-membered heterocycloalkyl group; R 3 H, R C , or R D And; R 4 , R 5 , R 6 , and R 7 These are H, OH, and OR, respectively, independently. C , OR D , R C , or R D and ； R 8 OH, NH 2 , OR C , OR D NHR C , or NR C R D And; R 9 H, OH, OR C , or OR D And; X is -S-, -S(O)-, or -SO 2 -and; R C is -LZ; R D is an arbitrarily substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 heteroalkyl, arbitrary C replaced 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted C 2 -C 6 Alkinyl, optionally substituted C 2 -C 6 Heteroalkynyl, arbitrarily substituted sic Roalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and is an arbitrarily substituted heteroaryl; L is an arbitrarily substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 heteroalkylene, Arbitrarily substituted C 2 -C 6 Alkenylene, optionally substituted C 2 -C 6 Heteroalkenylene, optional C replaced 2 -C 6 Alkynylene, optionally substituted C 2 -C 6 Heteroalkylene, arbitrarily placed Substituted cycloalkylene, arbitrarily substituted heterocycloalkylene, arbitrarily substituted Arylenes, or optionally substituted heteroarylenes; and Z is a reactive substituent present on L and present within the antibody or its antigen-binding fragment. It is a chemical site formed by a coupling reaction with the reactive substituent present. In the formula, Am is just one R C [Contains substituents].

152. The antibody or its antigen-binding fragment It is conjugated to the amatoxin via cysteine ​​residues in the Fc domain of the . , the conjugate according to claim 149.

153. The cysteine ​​residue is in the Fc domain of the antibody or its antigen-binding fragment. The conjugate according to claim 152, which is introduced by a mutation.

154. The cysteine ​​residue is selected from the group consisting of Cys118, Cys239, and Cys265. The conjugate described in item 153.

155. The cysteine ​​residue is naturally present in the Fc domain of the antibody or its antigen-binding fragment. The conjugate according to claim 152, which exists in the present.

156. The Fc domain is an IgG Fc domain, and the cysteine ​​residues are Cys261, Cys321, and Cys3 The conjugate according to claim 155, selected from the group consisting of 67 and Cys425.

157. R 1 However, H, OH, or OR A And; R 2 However, H, OH, or OR B And; R A and R B However, together with the oxygen atoms to which they are bonded, they combine to form the following: ： 【Chemistry 19】 ； R 3 , R 4 , R 6 , and R 7 However, each of them is H; R 5 is OR C And; R 8 However, OH or NH 2 and; as well R 9 The conjugate according to claim 150 or 151, wherein the conjugate is H or OH.

158. R 1 and R 2 However, each is independently either H or OH; R 3 R C And; R 4 , R 6 , and R 7 However, each of them is H; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The conjugate according to claim 150 or 151, wherein the conjugate is H or OH.

159. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 However, OR C , or R C And; R 5 However, H, OH, or OC 1 -C 6 It is alkyl; R 8 However, OH or NH 2 and; as well R 9 The conjugate according to claim 150 or 151, wherein the conjugate is H or OH.

160. R 1 and R 2 However, each is independently either H or OH; R 3 , R 6 , and R 7 However, each of them is H; R 4 and R 5 However, each is independently either H or OH; R 8 However, OR C or NHR C and; as well R 9 The conjugate according to claim 150 or 151, wherein the conjugate is H or OH.

161. The antibody or its antigen-binding fragment is taken up by CD135+ cells, The conjugate described in item 149.

162. The antibody or its antigen-binding fragment contains approximately 0.1 pM to approximately 1 μM of K d Combine with CD135 , the conjugate according to claim 149.

163. The antibody or its antigen-binding fragment is approximately 9 × 10 -2 M -1 s -1 ~Approx. 1×10 2 M -1 s -1 k on The conjugate according to claim 149, which is coupled to CD135.

164. The antibody or its antigen-binding fragment is a second antibody or its antigen-binding fragment of CD135. Competitively inhibits binding to the fragment of the second antibody or its antigen-binding fragment. However, the conjugate according to claim 149 includes the following complementarity determination region (CDR): a. CDR-H1 having the amino acid sequence SYYMH (SEQ ID NO: 1); b. CDR-H2 having the amino acid sequence IINPSGGSTSYAQKFQG (SEQ ID NO: 2); c. CDR having amino acid sequence GVGAHDAFDI (SEQ ID NO: 3) or VVAAAVADY (SEQ ID NO: 4) -H3； d. Amino acid sequence RSSQSLLHSNGNNYLD (SEQ ID NO: 5) or RSSQSLLHSNGYNYLD (SEQ ID NO: 6) ) CDR-L1; e. CDR-L2 having the amino acid sequence LGSNRAS (SEQ ID NO: 7); and f. CDR- having amino acid sequence MQGTHPAIS (SEQ ID NO: 8) or MQSLQTPFT (SEQ ID NO: 9) L3。

165. The antibody or its antigen-binding fragment is a second antibody or its antigen-binding fragment of CD135. Competitively inhibits binding to the fragment of the second antibody or its antigen-binding fragment. However, the conjugate according to claim 149 includes the following complementarity determination region (CDR): a. CDR-H1 having the amino acid sequence SYAIS (SEQ ID NO: 10); b. CDR-H2 having the amino acid sequence GIIPIFGTANYAQKFQG (SEQ ID NO: 11); c. CDR-H3 having the amino acid sequence FALFGFREQAFDI (SEQ ID NO: 12); d. CDR-L1 having the amino acid sequence RASQSISSYLN (SEQ ID NO: 13); e. CDR-L2 having the amino acid sequence AASSLQS (SEQ ID NO: 14); and f. CDR-L3 having the amino acid sequence QQSYSTPFT (SEQ ID NO: 15).

166. The antibody or its antigen-binding fragment is a monoclonal antibody or its antigen-binding Fragments, polyclonal antibodies or their antigen-binding fragments, humanized antibodies or The antigen-binding fragment, the bispecific antibody or its antigen-binding fragment, bivariable Immunoglobulin domain, single-stranded Fv molecule (scFv), diabody, triabody, nanobody D, antibody-like protein scaffold, Fv fragment, Fab fragment, F(ab') 2 molecules, and The conjugate according to claim 149, selected from the group consisting of tandem scFV.

167. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or The antigen-binding fragment is a conjugate in which Cy is Pseudomonas exotoxin A. 。

168. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, a conjugate in which Cy is deBouganin.

169. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, and Cy is the diphtheria toxin, forming a conjugate.

170. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, and the Cy component is a saporin, forming a conjugate.

171. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or The antigen-binding fragment is such that Cy is maytansine or maytansinoid. Njugate.

172. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, a conjugate in which Cy is auristatin.

173. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, and the Cy is an anthracycline, making it a conjugate.

174. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, a conjugate in which Cy is calicheamycin.

175. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, a conjugate in which Cy is irinotecan.

176. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is the antigen-binding fragment, a conjugate in which Cy has SN-38.

177. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, a conjugate in which Cy is duocalmycin.

178. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, and Cy is pyrrolobenzodiazepine or pyrrolobenzodia A conjugate, which is a zepine dimer.

179. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD45 or It is an antigen-binding fragment, and Cy is indolinobenzodiazepine or indolino A conjugate, which is a dimer of zodiazepines.

180. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or The antigen-binding fragment is a conjugate in which Cy is Pseudomonas exotoxin A. 。

181. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, a conjugate in which Cy is deBouganin.

182. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, and Cy is the diphtheria toxin, forming a conjugate.

183. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, and the Cy component is a saporin, forming a conjugate.

184. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or The antigen-binding fragment is such that Cy is maytansine or maytansinoid. Njugate.

185. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, a conjugate in which Cy is auristatin.

186. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, and the Cy is an anthracycline, making it a conjugate.

187. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, a conjugate in which Cy is calicheamycin.

188. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, a conjugate in which Cy is irinotecan.

189. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is the antigen-binding fragment, a conjugate in which Cy has SN-38.

190. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, a conjugate in which Cy is duocalmycin.

191. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, and Cy is pyrrolobenzodiazepine or pyrrolobenzodia A conjugate, which is a zepine dimer.

192. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD135 or It is an antigen-binding fragment, and Cy is indolinobenzodiazepine or indolino A conjugate, which is a dimer of zodiazepines.

193. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or The antigen-binding fragment is a conjugate in which Cy is Pseudomonas exotoxin A. 。

194. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, a conjugate in which Cy is deBouganin.

195. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, and Cy is the diphtheria toxin, forming a conjugate.

196. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, and the Cy component is a saporin, forming a conjugate.

197. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or The antigen-binding fragment is such that Cy is maytansine or maytansinoid. Njugate.

198. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, a conjugate in which Cy is auristatin.

199. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, and the Cy is an anthracycline, making it a conjugate.

200. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, a conjugate in which Cy is calicheamycin.

201. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, a conjugate in which Cy is irinotecan.

202. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is the antigen-binding fragment, a conjugate in which Cy has SN-38.

203. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, a conjugate in which Cy is duocalmycin.

204. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, and Cy is pyrrolobenzodiazepine or pyrrolobenzodia A conjugate, which is a zepine dimer.

205. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD34 or It is an antigen-binding fragment, and Cy is indolinobenzodiazepine or indolino A conjugate, which is a dimer of zodiazepines.

206. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or The antigen-binding fragment is a conjugate in which Cy is Pseudomonas exotoxin A. 。

207. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, a conjugate in which Cy is deBouganin.

208. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, and Cy is the diphtheria toxin, forming a conjugate.

209. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, and the Cy component is a saporin, forming a conjugate.

210. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or The antigen-binding fragment is such that Cy is maytansine or maytansinoid. Njugate.

211. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, a conjugate in which Cy is auristatin.

212. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, and the Cy is an anthracycline, making it a conjugate.

213. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, a conjugate in which Cy is calicheamycin.

214. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, a conjugate in which Cy is irinotecan.

215. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is the antigen-binding fragment, a conjugate in which Cy has SN-38.

216. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, a conjugate in which Cy is duocalmycin.

217. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, and Cy is pyrrolobenzodiazepine or pyrrolobenzodia A conjugate, which is a zepine dimer.

218. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD90 or It is an antigen-binding fragment, and Cy is indolinobenzodiazepine or indolino A conjugate, which is a dimer of zodiazepines.

219. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or The antigen-binding fragment is a conjugate in which Cy is Pseudomonas exotoxin A. 。

220. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, a conjugate in which Cy is deBouganin.

221. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, and Cy is the diphtheria toxin, forming a conjugate.

222. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, and the Cy component is a saporin, forming a conjugate.

223. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or The antigen-binding fragment is such that Cy is maytansine or maytansinoid. Njugate.

224. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, a conjugate in which Cy is auristatin.

225. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, and the Cy is an anthracycline, making it a conjugate.

226. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, a conjugate in which Cy is calicheamycin.

227. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, a conjugate in which Cy is irinotecan.

228. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is the antigen-binding fragment, a conjugate in which Cy has SN-38.

229. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, a conjugate in which Cy is duocalmycin.

230. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, and Cy is pyrrolobenzodiazepine or pyrrolobenzodia A conjugate, which is a zepine dimer.

231. A conjugate represented by the formula Ab-Cy, wherein Ab is an antibody that binds to CD110 or It is an antigen-binding fragment, and Cy is indolinobenzodiazepine or indolino A conjugate, which is a dimer of zodiazepines.

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