Methods for recruiting hematopoietic progenitor cells

Abstract

A method is provided for recruiting hematopoietic progenitor cells from the bone marrow of a target to the peripheral blood. The method comprises (a) administering an effective dose of natalizumab to the target, and (b) administering an effective dose of a CXCR4 inhibitor to the target, at least 48 hours after step (a). The use of such recruitment procedures in cell collection and therapy is also provided.

Description

Technical Field

[0001] Related applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 454,684, filed Mar. 26, 2023, the content of which is hereby incorporated by reference in its entirety.

[0002] Sequence listing The XML titled 99455.xml, created on Mar. 25, 2024 and consisting of 215,618 bytes, filed simultaneously with the filing of this application, is hereby incorporated by reference.

[0003] Field of Invention In some embodiments, the present invention relates to a method for mobilizing hematopoietic progenitor cells.

Background Art

[0004] Hematopoietic cell transplantation is an important treatment for many hematologic malignancies and solid tumors, and in many cases a life-saving treatment, as a means to reconstitute blood cells after high-dose chemotherapy. The gold standard for mobilization typically includes the use of granulocyte colony-stimulating factor (G-CSF) to mobilize peripheral blood stem cells (PBSC), which has largely replaced bone marrow (BM) as a source of stem cells for both autologous and allogeneic cell transplantation. Despite the increased number of CD34+ stem cells obtained after G-CSF mobilization compared to BM harvest, a minimum number of CD34 / kg (about ≧2×10 6It is necessary to obtain (individual cells). Conversely, the SDF-1 / CXCR4 interaction plays an important role in the resting state and retention of HSCs in the bone marrow. Plerixafor (Mozobil, AMD3100) is a bicyclum molecule that promotes HSC recruitment by reversibly inhibiting the binding of SDF-1 to CXCR4. Plerixafor is approved for use in combination with G-CSF for stem cell recruitment in myeloma and lymphoma patients.

[0005] Motixafortide (4F-benzoyl-TN14003, Rehovot, Israel, Biokine Therapeutics) (T-140), also known as BKT 140, is a highly selective CXCR4 antagonist originally designed to inhibit the binding of human immunodeficiency virus (HIV) to CXCR4. In mice, this drug induced up to a 10-fold increase in PB progenitor cells, peaking 1-2 hours after dose administration. BKT 140, acting synergistically with G-CSF, increased PB progenitor cells 78-fold compared to controls, which was higher than that observed with the combination of plerixafor and G-CSF (Abraham et al. Stem Cells. 2007;25:2158-66). In a Phase I / IIA dose-escalation study in myeloma patients, BTK 140 was well-tolerated and dose-dependently increased the mean absolute number of PB CD34+ cells. At the highest dose tested (900 μg / kg), the mean count of PB CD34+ cells was 20.6 × 10⁶. 6 The number of cells / kg decreased, and the number of required apheresis procedures was reduced from 2.25 at low doses to 1 (Nagler et al. ASH Annual Meeting Abstracts; 2010. p. 2260).

[0006] Integrins are a family of structurally and functionally diverse transmembrane glycoproteins that mediate cell-cell and cell-matrix interactions in a wide range of biological contexts. Vertebrates possess 18 distinct α-subunits and 8 distinct β-subunits, resulting in 24 distinct non-covalent αβ heterodimers that can bind to a wide variety of ligands. One such heterodimer expressed in hematopoietic stem cells, α4β1, is called late-stage antigen 4 (VLA-4) and mediates the adhesion of HSCs to vascular cell adhesion molecule-1 (VCAM-1) in the bone marrow stroma. In preclinical studies, administration of anti-VLA-4 antibodies recruited HSC progenitor cells into the bloodstream (Vermeulen et al. Blood. 1998;92:894-900).

[0007] Natalizumab, a recombinant humanized monoclonal antibody against the α4 subunit of VLA-4, approved for the treatment of multiple sclerosis (MS) and Crohn's disease, has been shown to increase peripheral blood CD34+ cells in patients with relapsing-remitting MS. Zohren et al. showed a gradual increase in circulating CD34+ cells in MS patients, with a peak concentration of 10.4 cells / μL 72 hours after natalizumab administration (Zohren et al. Blood. 2008;111:3893-5). Jing et al. demonstrated a 7-fold increase in PB CD34+ cells and a dose-dependent 7-fold increase in BM CD34+ cells in MS patients treated with natalizumab, reaching a peak absolute count on day 4 post-treatment (Jing et al. 2010;45:1489-96). Furthermore, simultaneous blockade of VLA-4 and CXCR4 has been shown to have a greater additive effect on primate stem cell recruitment compared to either drug alone (Bonig et al. Stem Cells. 2009;27:836-7). Unfortunately, the increase in PB CD34+ cells induced by natalizumab persists for at least one month after drug administration, limiting its use in healthy donors (Zohren and Bonig, op. cit.).

[0008] Despite numerous advances, there is still a need for novel therapeutic modalities that lead to the most effective and efficient stem cell mobilization strategies, particularly in patients at risk of mobilization failure.

[0009] Further background technologies include: U.S. Patent No. 8,455,450 International Publication No. 2018 / 085574 PMID:24476957 [Overview of the project]

[0010] According to one aspect of several embodiments of the present invention, a method for recruiting hematopoietic progenitor cells from a target bone marrow to peripheral blood, (a) the step of administering an effective dose of natalizumab to the target, and A method is provided which includes (b) administering an effective amount of CXCR4 inhibitor to a subject, at least 48 hours after step (a).

[0011] According to one aspect of several embodiments of the present invention, a method for obtaining a therapeutically effective amount of hematopoietic progenitor cells from a subject, (a) A step of administering an effective dose of natalizumab to the target, (b) a step of administering an effective amount of CXCR4 inhibitor to the subject, which is performed at least 48 hours after step (a), and (c) A method is provided which includes the step of collecting hematopoietic progenitor cells by apheresis.

[0012] According to one aspect of several embodiments of the present invention, a combination of effective doses of natalizumab and a CXCR4 inhibitor for use in the recruitment of hematopoietic progenitor cells in a subject is provided, wherein the CXCR4 inhibitor is administered to the subject at least 48 hours after the administration of natalizumab.

[0013] According to one aspect of some embodiments of the present invention, a combination of an effective amount of natalizumab and a CXCR4 inhibitor for use in a method of collecting hematopoietic progenitor cells by apheresis, the method comprising mobilizing hematopoietic progenitor cells, the hematopoietic progenitor cells being subsequently collected by apheresis, and the CXCR4 inhibitor being administered to the subject at least 48 hours after the administration of natalizumab, is provided.

[0014] According to some embodiments of the present invention, the subject is a human subject.

[0015] According to some embodiments of the present invention, the subject is a donor of hematopoietic progenitor cells.

[0016] According to some embodiments of the present invention, the subject is diagnosed with cancer.

[0017] According to some embodiments of the present invention, the hematopoietic progenitor cells are for autologous transplantation.

[0018] According to some embodiments of the present invention, the hematopoietic progenitor cells are for allogeneic transplantation.

[0019] According to some embodiments of the present invention, the effective amount comprises repeated cycles of administration of natalizumab and the CXCR4 inhibitor.

[0020] According to some embodiments of the present invention, the at least 48 hours includes up to 96 hours. <00000�5> According to some embodiments of the present invention, the at least 48 hours includes up to 72 hours.

[0022] According to some embodiments of the present invention, the CXCR4 inhibitor is a peptide, small molecule, antibody, nucleic acid, or a combination thereof.

[0023] According to some embodiments of the present invention, the CXCR4 inhibitor is the peptide set forth in SEQ ID NO: 1.

[0024] According to some embodiments of the present invention, the CXCR4 inhibitor is a small molecule.

[0025] According to some embodiments of the present invention, the CXCR4 inhibitor is AMD3100.

[0026] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, exemplary methods and / or materials are described below. In case of conflict, the present patent specification, including definitions, will prevail. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Embodiments for Carrying Out the Invention

[0027] In some embodiments, the present invention relates to a method for mobilizing hematopoietic progenitor cells.

[0028] Before detailing at least one embodiment of the present invention, it is to be understood that the present invention is not necessarily limited in its application to the details described in the following description or illustrated by the examples. The present invention is capable of other embodiments or of being practiced or carried out in various ways.

[0029] The inventors unexpectedly discovered that by determining the timing of administration of the CXCR4 inhibitor to be at least 48 hours after administration of natalizumab, it becomes possible to obtain a considerably large amount of hematopoietic progenitor cells.

[0030] Thus, according to one aspect of the present invention, a method for mobilizing hematopoietic progenitor cells from the bone marrow to the peripheral blood of a subject, comprising: (a) administering an effective amount of natalizumab to the subject, and A method is provided which includes (b) administering an effective amount of CXCR4 inhibitor to a subject, at least 48 hours after step (a).

[0031] In addition or alternative embodiments, a method for obtaining a therapeutically effective amount of hematopoietic progenitor cells from a subject, (a) A step of administering an effective dose of natalizumab to the target, (b) a step of administering an effective amount of CXCR4 inhibitor to the subject, which is performed at least 48 hours after step (a), and (c) A method is provided which includes the step of collecting hematopoietic progenitor cells by apheresis.

[0032] In an additional or alternative embodiment, a combination of effective doses of natalizumab and a CXCR4 inhibitor is provided for use in the recruitment of hematopoietic progenitor cells in a subject, wherein the CXCR4 inhibitor is administered to the subject at least 48 hours after the administration of natalizumab.

[0033] In an additional or alternative embodiment, a combination of effective doses of natalizumab and a CXCR4 inhibitor for use in a method of collecting hematopoietic progenitor cells by apheresis is provided, wherein the method comprises mobilizing hematopoietic progenitor cells, the hematopoietic progenitor cells being subsequently collected by apheresis, and the CXCR4 inhibitor being administered to the subject at least 48 hours after administration of natalizumab.

[0034] As used herein, the term “hematopoietic progenitor cells” refers to a subset of cells that can be found in the bone marrow (BM) niche. These cells are characterized by pluripotency, which allows them to self-replicate and produce mature blood cells such as erythrocytes, leukocytes, platelets, and lymphocytes. These cells typically include hematopoietic stem cells (HSCs) that express CD34 and are referred to as CD34+ cells. CD34 is a marker for human HSCs, and all colony-forming activity of human bone marrow (BM) cells is found in the CD34+ fraction.

[0035] Hematopoietic progenitor cells in their native form after apheresis, or modified (e.g., differentiated in culture, grown in culture) hematopoietic progenitor cells, are typically used to treat a variety of medical conditions.

[0036] As used herein, the term “mobilization” refers to the release of hematopoietic progenitor cells (e.g., hematopoietic stem cells) from the bone marrow into the peripheral blood circulation.

[0037] As used herein, “increasing recruitment” means inducing the recruitment of peripheral blood progenitor cells, increasing the circulating level of HSCs, or enhancing or promoting hematopoietic reconstitution or engraftment in subjects where it is needed.

[0038] According to one embodiment, hematopoietic stem cell recruitment is increased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, or 200% compared to recruitment in the absence of the agents of the present invention (i.e., CXCR4 inhibitors and natalizumab). According to one embodiment, hematopoietic progenitor cell recruitment is increased by at least about 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold compared to recruitment in the absence of the agents of the present invention. The measurement of enhanced hematopoietic progenitor cell recruitment is known to those skilled in the art.

[0039] As used herein, the term “subject” refers to a mammalian subject of any sex and age, such as a human subject. A subject may be a healthy subject (e.g., also referred to as a “donor subject”) who is to serve as a donor for hematopoietic progenitor cell transplantation. Alternatively, a subject may have a disease or condition (e.g., a malignant disease, or a non-malignant disease such as immunodeficiency) that requires stem cell mobilization or transplantation (i.e., autologous or from a donor, i.e. non-autologous, i.e. allogeneic). In the latter case, the subject is a recipient requiring stem cell transplantation.

[0040] As used herein, the term “healthy subject” refers to a subject who has not been diagnosed with a disease or disorder to which hematopoietic progenitor cell transplantation is applicable. According to specific embodiments, the healthy subject is not suffering from a hematopoietic disorder or malignant tumor.

[0041] As used herein, the term “CXCR4 inhibitor” refers to molecules and compositions that interfere with or inhibit the biological activity of the CXCR4 receptor. The biological activity of the CXCR4 receptor may include the recruitment of stem cells into the bloodstream and the entry of viruses into cells or the replication of viruses within cells.

[0042] CXCR4 inhibitors can encompass a wide range of chemical molecules, including small organic or inorganic molecules, polysaccharides, biomacromolecules such as peptides, proteins, peptide analogs and derivatives, peptide mimes, antibodies, antibody fragments, nucleic acids, aptamers and other nucleic acid analogs and derivatives, extracts made from biomolecules such as bacteria, plants, fungi, or animal cells or tissues, naturally occurring compositions, or synthetic compositions.

[0043] While we do not wish to be constrained by theory, CXCR4 inhibitors may act through several different pathways. For example, CXCR4 inhibitors may bind to ligand-binding sites on the CXCR4 receptor to prevent ligand binding to the CXCR4 receptor, bind to non-ligand-binding sites on the CXCR4 receptor to prevent ligand binding to the CXCR4 receptor, bind to the CXCR4 receptor ligand to prevent ligand binding to the CXCR4 receptor, or inhibit the expression of polynucleotides (e.g., mRNA) that express CXCR4.

[0044] In some embodiments, the CXCR4 inhibitor inhibits the biological activity of the CXCR4 receptor by at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% compared to a control. In some embodiments, the CXCR4 inhibitor completely abolishes the biological activity of the CXCR4 receptor compared to a control. The control may include a sample that has not been treated with the inhibitor.

[0045] In some embodiments, the CXCR4 inhibitor is a nucleic acid. Exemplary nucleic acid inhibitors of CXCR4 include, but are not limited to, antisense oligonucleotides, siRNA, shRNA, microRNA, aptamers, ribozymes, and decoy oligonucleotides. Nucleic acid inhibitors of CXCR4 can inhibit the expression of the CXCR4 gene.

[0046] Exemplary anti-CXCR4 siRNAs are described, for example, in U.S. Patent Application Publication No. 2007 / 0238868 and U.S. Patent Application Publication No. 2009 / 0253772, the contents of which are incorporated herein by reference. Several exemplary CXCR4 antisense oligonucleotides are described, for example, in U.S. Patent Application Publication No. 2004 / 0209837, the contents of which are incorporated herein by reference.

[0047] In some embodiments, the CXCR4 inhibitor binds to CXCR4 or CXCL12 (SDF-1 alpha). In other embodiments, the CXCR4 inhibitor is an antibody or antibody fragment. In some embodiments, the CXCR4 inhibitor is a small molecule, e.g., AMD-3100, ALX40-4C, T22, T140, Met-SDFl beta, T134, or AMD-3465.

[0048] Exemplary CXCR4 inhibitors include, but are not limited to, 2,2'-bisicram, 6,6'-bisicram, embodiments described in U.S. Patent Nos. 5,021,409 and 6,001,826, and 1,1'-[1,4-phenylene-bis(methylene)]-bis-1,4,8,11tetraazacyclotetradecane, particularly described in U.S. Patent No. 5,583,131 and designated herein as AMD3100. In some embodiments, the CXCR4 inhibitor may be N'-(1H-benzimidazole-2-ylmethyl)-N'-(5,6,7,8-tetrahydroquinoline-8-yl)-butan-1,4-diamine, CTCF-0214, CTCF-9908, CP-1221 (linear peptides, cyclic peptides, natural amino acids, unnatural amino acids, and peptide mimetic compounds), 4F-benzoyl TN24003, KRH-1120, KRH-1636, KRH-2731, polyfemusin analogs, ALX40-4C, or those described in International Publications 01 / 85196, 99 / 50461, 01 / 94420, and 03 / 090512 (each of which is incorporated herein by reference in whole).

[0049] In some embodiments, the CXCR4 inhibitor is a T-140 analog and antibody as described in U.S. Patent Application Publication No. 2010 / 0055088, and a cycle polyamine as described in U.S. Patent Application Publication No. 2009 / 0221683. This includes compounds disclosed in U.S. Patent Publication Nos. 2004 / 0209921, 2005 / 0059702, 2005 / 0043367, 2005 / 0277670, 2010 / 0178271, and 2003 / 0220341, U.S. Patent Nos. 5,021,409, 6,001,826, and 5,583,131, and International Publication No. 03 / 011277, each of which is incorporated herein by reference in whole.

[0050] CXCR4 inhibitors may include, but are not limited to, polypeptides that specifically bind to CXCR4. Such inhibitors include T140 and derivatives of T140. Exemplary derivatives of T140 include, but are not limited to, TN14003, TC14012, and TE14011, as well as derivatives found in Tamamura, H. et al. Org. Biomol. Chem. 1:3656-3662, 2003, which is incorporated herein by reference in whole.

[0051] In specific embodiments, the CXCR4 antagonist peptide of the present invention is, for example, a 4F-benzoyl-TN14003 (SEQ ID NO: 1) analog and derivative, and is structurally and functionally related to the peptide disclosed in International Publication Nos. 2002 / 020561 and International Publication Nos. 2004 / 020462, which are also known as the "T-140 analog," as detailed below.

[0052] In various specific embodiments, the T-140 analog or T-140 derivative has the amino acid sequence or a salt thereof described in the following formula (I). [ka] During the ceremony, A1 is either an arginine residue, a lysine residue, an ornithine residue, a citrulline residue, an alanine residue, or a glutamate residue, or an N-α substituted derivative of any of these amino acids, or A1 is absent. A2 represents either an arginine residue or a glutamate residue if A1 is present, or if A1 is not present, A2 represents either an arginine residue or a glutamate residue, or an N-α substituted derivative of these amino acids. A3 represents an aromatic amino acid residue. A4, A5, and A9 each independently represent an arginine residue, a lysine residue, an ornithine residue, a citrulline residue, an alanine residue, or a glutamate residue. A6 represents a proline residue, glycine residue, ornithine residue, lysine residue, alanine residue, citrulline residue, arginine residue, or glutamate residue. A7 represents a proline residue, glycine residue, ornithine residue, lysine residue, alanine residue, citrulline residue, or arginine residue. A8 represents a tyrosine residue, a phenylalanine residue, an alanine residue, a naphthylalanine residue, a citrulline residue, or a glutamic acid residue. A 10 This represents a citrulline residue, a glutamate residue, an arginine residue, or a lysine residue. A 11 represents an arginine residue, glutamic acid residue, lysine residue, or citrulline residue, and the C-terminal carboxyl may be derivatized. Furthermore, the cysteine ​​residue at position 4 or 13 can form a disulfide bond, and the amino acid can be either L-type or D-type.

[0053] An exemplary peptide following formula (I) is a peptide having the amino acid sequence described in any one of the sequence numbers 1 to 72 shown in Table 1 below.

[0054] [Table 1] TIFF2026513170000003.tif183129TIFF2026513170000004.tif192133TIFF2026513170000005.tif88131

[0055] In a specific embodiment, in each of Sequence IDs 1 to 72, two cysteine ​​residues are linked by a disulfide bond.

[0056] In another embodiment, the analog or derivative has the amino acid sequence described in Sequence ID No. 65 (H-Arg-Arg-Nal-Cys-Tyr-Cit-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH, TC14003).

[0057] In another embodiment, the peptide used in the composition and method of the present invention essentially consists of the amino acid sequence described in SEQ ID NO: 1. In another embodiment, the peptide used in the composition and method of the present invention contains the amino acid sequence described in SEQ ID NO: 1. In another embodiment, the peptide is at least 60%, at least 70%, or at least 80% homologous to SEQ ID NO: 1. In another embodiment, the peptide is at least 90% homologous to SEQ ID NO: 1. In another embodiment, the peptide is at least about 95% homologous to SEQ ID NO: 1. Each possibility represents a separate embodiment of the present invention.

[0058] In various other embodiments, the peptide is selected from SEQ ID NOs: 1 to 72, and each possibility represents a separate embodiment of the present invention.

[0059] In another embodiment, the peptide has the amino acid sequence described in any one of SEQ ID NOs: 1-4, 10, 46, 47, 51-56, 65, 66, 68, 70, and 71. In another embodiment, the peptide has the amino acid sequence described in any one of SEQ ID NOs: 4, 10, 46, 47, 68, and 70. In another embodiment, the peptide has the amino acid sequence described in any one of SEQ ID NOs: 1, 2, 51, 65, and 66. In another embodiment, the peptide has the amino acid sequence described in any one of SEQ ID NOs: 53-56.

[0060] In one embodiment, the peptide has the amino acid sequence described in SEQ ID NO: 1. In another embodiment, the peptide has the amino acid sequence described in SEQ ID NO: 2. In yet another embodiment, the peptide has the amino acid sequence described in SEQ ID NO: 51. In yet another embodiment, the peptide has the amino acid sequence described in SEQ ID NO: 66.

[0061] Other CXCR4 peptide inhibitors (antagonists) include, but are not limited to, LY2510924 (Lilly Oncology), CTCE-9908 (Huang et al. 2009 Journal of Surgical Research 155:231-236), and the Fc131 analogs and nanobodies described in the following references (each of which is incorporated herein by reference in its entirety). Tan NC, Yu P, Kwon YU, Kodadek T. High-throughput evaluation of relative cell permeability between peptoids and peptides. Bioorg Med Chem. 2008;16:5853-61. Kwon YU, Kodadek T. Quantitative evaluation of the relative cell permeability of peptoids and peptides. J Am Chem Soc. 2007;129:1508. Miller S, Simon R, Ng S, Zuckermann R, Kerr J, Moos W. Comparison of the proteolytic susceptibilities of homologous L-amino acid, D-amino acid, and N-substituted glycine peptide and peptoid oligomers. Drug Dev Res. 1995;35:20-32. Yoshikawa Y, Kobayashi K, Oishi S, Fujii N, Furuya T. Molecular modeling study of cyclic pentapeptide CXCR4 antagonists: new insight into CXCR4-FC131 interactions. Bioorg Med Chem Lett. 2012;22:2146-50. Jaahnichen S, Blanchetot C, Maussang D, Gonzalez-Pajuelo M, Chow KY, Bosch L, De Vrieze S, Serruys B, Ulrichts H, Vandevelde W. CXCR4 nanobodies (VHH-based single variable domains) potently inhibit chemotaxis and HIV-1 replication and mobilized stem cells. Proc Natl Acad Sci USA. 2010;107:20565-70.

[0062] In a specific embodiment, the peptide, under the name mothixafortide, has the amino acid sequence described in Sequence ID No. 1.

[0063] As mentioned above, the VLA-4 inhibitor (VLA4i) drug is natalizumab [abbreviated as (N) herein].

[0064] According to some embodiments, natalizumab is marketed under the trade name Tysabri®.

[0065] According to some embodiments, natalizumab is marketed under the trade name Antegren®.

[0066] According to specific embodiments, the peptide of the present invention (e.g., BL-8040) or a pharmaceutical composition containing the same is administered in doses ranging from 0.1 to 10 mg / kg body weight, 0.1 to 2 mg / kg body weight, 0.1 to 1 mg / kg body weight, 0.3 to 10 mg / kg body weight, and 0.3 to 2 mg / kg body weight.

[0067] In a specific embodiment, BL-8040 is administered at a dose of 1-2 mg / kg body weight.

[0068] In a specific embodiment, BL-8040 is administered at a dose of 1.25 to 1.5 mg / kg body weight.

[0069] In a specific embodiment, BL-8040 is administered at a dose of 1.25 mg / kg body weight.

[0070] In a specific embodiment, BL-8040 is administered subcutaneously (SC).

[0071] As mentioned above, CXCR4 inhibitors (e.g., BL-8040) are administered at least 48 hours after natalizumab administration.

[0072] In specific embodiments, at least 48 hours include a maximum of 96 hours (e.g., 48 to 96 hours).

[0073] In specific embodiments, at least 48 hours include a maximum of 80 hours (e.g., 48 to 80 hours).

[0074] In specific embodiments, at least 48 hours include a maximum of 72 hours (e.g., 48 to 72 hours).

[0075] In specific embodiments, at least 48 hours includes approximately 48 hours (for example, 48 hours).

[0076] In a specific embodiment, the method includes performing apheresis to recover hematopoietic progenitor cells from the target peripheral blood after drug administration.

[0077] According to some embodiments of the present invention, the subject undergoes apheresis approximately 4 hours to a maximum of approximately 12 hours after administration of the CXCR4 inhibitor.

[0078] Methods for collecting peripheral blood are well known in the art and include, but are not limited to, collecting whole blood (up to 500 ml) from a subject and collecting it in a container with an anticoagulant (e.g., heparin or citrate), and apheresis.

[0079] As used herein, the term “apheresis” refers to the procedure of passing an individual’s peripheral blood through a device to obtain its major components (e.g., HSCs) and returning other components to the subject’s circulation. Apheresis is generally a three-step process involving (1) drawing blood from the subject, (2) separating the blood components (e.g., based on density), and (3) returning specific components of the blood to the subject by transfusion. Blood is typically separated into three fractions: red blood cells (about 45% of total blood), “buffy coat” (less than 1% of total blood), and plasma (about 55% of total blood). Depending on the components of the blood to be extracted, various types of apheresis procedures can be used.

[0080] To confirm that hematopoietic stem cells have been recruited into the peripheral blood, any method, including quantitative and qualitative methods, may be used. The method typically involves taking a certain amount of the patient's blood and analyzing the number of cells in the blood. The number of cells can be analyzed using any method, which includes, but is not limited to, ELISA, FACS analysis, Coulter counters and other hematological counting devices, morphological identification, and PCR for identifying a given cell type. Cells can be identified by any method known to those skilled in the art, including, but not limited to, identification of one or more proteins specifically expressed by progenitor cells, morphological identification, mRNA expression identification, and PCR identification. Cell identification can be performed at any point in time after administration of the drug (a CXCR4 inhibitor administered after natalizumab), including, but not limited to, 5 minutes, 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 12 hours, 1 day, and 2 days.

[0081] According to one embodiment, after collection (and optionally isolation), hematopoietic stem cells can be preserved (for example, for later use), grown in culture, or administered to subjects requiring them (as will be discussed in detail below herein). Thus, the method of recruiting stem cells can be used to recruit progenitor hematopoietic cells in individuals that will serve as allogeneic or autologous donors of hematopoietic stem cells.

[0082] To obtain a sufficient number of hematopoietic progenitor cells for transplantation, the collected hematopoietic stem cells may be further cultured in vitro to increase their proliferation.

[0083] During in vitro culture, the cellular phenotype of hematopoietic progenitor cells may change. The presence or absence of antigens on the cell surface can be analyzed using specific antibodies (e.g., CD34 antibodies) by methods well known to those skilled in the art, such as FACS ELISA. The presence or absence of antigens can further be analyzed by other methods well known in the art, such as RT-PCR or similar methods.

[0084] In addition, hematopoietic progenitor cells can also be used in gene therapy. Because pluripotent hematopoietic stem cells possess self-renewal capabilities and produce both mature blood cells and blood cell progenitor cells, hematopoietic progenitor cells are suitable targets for gene therapy. After collection (and selective isolation), hematopoietic progenitor cells can be modified to deliver gene products upon reintroduction into an organism. After modification, the cells are reinjected into subjects requiring such treatment.

[0085] According to another aspect of the present invention, an isolated population of hematopoietic progenitor cells is provided, which can be obtained by the methods of some embodiments of the present invention.

[0086] As used herein, the term “isolated cell population” refers to cells isolated from their natural environment (e.g., the human body).

[0087] According to one embodiment, the population may include CXCr4 inhibitors and / or natalizumab.

[0088] According to one embodiment, cells are cultured ex vivo or in vitro.

[0089] Each drug can be administered to a subject either on its own or as part of a pharmaceutical composition that includes a physiologically acceptable carrier. The purpose of the pharmaceutical composition is to facilitate the administration of the active ingredient to the organism.

[0090] It will be understood that the pharmaceutical composition may further contain other compounds, such as those detailed above.

[0091] As used herein, “pharmaceutical composition” refers to a preparation of one or more of the active ingredients described herein with other chemical components such as physiologically suitable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of a compound to a living organism.

[0092] In this specification, the term "active ingredient" refers to a drug that affects mobilization.

[0093] In the following, the terms "physiologically acceptable carrier" and "pharmaceutically acceptable carrier," which may be used interchangeably, refer to carriers or diluents that do not cause significant irritation to the organism and do not impair the biological activity and properties of the administered compound. These terms include adjuvants.

[0094] In this specification, the term “excipient” refers to an inert substance added to a pharmaceutical composition to further facilitate the administration of the active ingredient. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol.

[0095] Techniques for the formulation and administration of drugs can be found in the latest edition of “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, PA, which is incorporated herein by reference.

[0096] Dosage and administration intervals can be individually adjusted to obtain a sufficient level of the active ingredient (minimum effective concentration, MEC) to induce or inhibit the biological effect. While MEC varies from preparation to preparation, it can be estimated from in vitro data. The dose required to achieve MEC depends on individual characteristics and administration route. Plasma concentrations can be determined using detection assays.

[0097] The agents of the present invention may be supplied in packs or dispenser devices, such as FDA-approved kits, which may optionally contain one or more unit dosage forms containing the active ingredient. Packs may include, for example, metal foil or plastic foil, such as blister packs. Packs or dispenser devices may be accompanied by instructions for administration. Packs or dispensers may also be fitted with a notice attached to the container in the format prescribed by the government agency that regulates the manufacture, use, or sale of pharmaceuticals, which reflects that the form of the composition or its administration to humans or veterinary use has been approved by the government agency. Such notices may be, for example, a label approved by the U.S. Food and Drug Administration for prescription drugs or an approved product insert. Compositions containing the preparations of the present invention formulated in a suitable pharmaceutical carrier may also be prepared, placed in appropriate containers, and labeled with indications for the treatment of the condition to which they are indicated, as further detailed above.

[0098] Typical conditions that can be alleviated or otherwise benefit from enhanced survival and / or proliferation and / or recruitment of hematopoietic stem cells, and / or hematopoietic progenitor cell transplantation include, but are not limited to, hematopoietic disorders and malignancies, e.g., aplastic anemia, lymphoma, leukemia, immunodeficiency, severe combined immunodeficiency (SCID), cytopenia (e.g., anemia, leukopenia, neutropenia, thrombocytopenia, granulocytopenia, pancytopenia), drug-induced cytopenia, toxin-induced cytopenia, radiation-induced cytopenia, cytopenia associated with conventional bone marrow transplantation, hematopoietic dysfunction associated with oncological treatment, chemotherapy, or radiotherapy, anemia or cytopenia associated with chronic disease (also referred to as anemia or cytopenia associated with chronic inflammation), osteopetrosis, Gaucher disease, thalassemia, and other congenital or genetically determined hematopoietic disorders.

[0099] According to one embodiment, the method of the present invention is useful in increasing the success of transplantation during and after immunosuppressive therapy, as well as in providing more efficient wound healing and treatment of bacterial inflammation.

[0100] According to one embodiment, the method of the present invention is useful for treating subjects in an immunocompromised state or subjects whose immune system is otherwise impaired. Typical conditions that are alleviated or otherwise benefit from the method of the present invention include, but are not limited to, subjects infected with retroviruses, more specifically, subjects infected with human immunodeficiency virus (HIV).

[0101] Therefore, the method of the present invention can be used to treat a wide range of conditions in which it is beneficial to increase the level of hematopoietic progenitor cells in a subject, or to harvest progenitor cells for subsequent stem cell transplantation. The compound is also administered to regenerate myocardium by mobilizing bone marrow progenitor cells.

[0102] The methods described herein are particularly suitable for subjects requiring repeated or high-dose chemotherapy. In some cancer patients, the opportunity to increase the dose of chemotherapy or to complete prescribed chemotherapy is often limited by hematopoietic toxicity. Repeated or high-dose cycles of chemotherapy can cause severe progenitor cell depletion, which can lead to long-term, serious hematopoietic sequelae and bone marrow exhaustion. When used in combination with chemotherapy, the methods of the present invention result in improved cell viability, blood cell reconstitution, and hematological counts.

[0103] In one embodiment, the disease or condition is a malignant disease.

[0104] As used herein, the terms “malignant disease” or “cancer” refer to any cancerous disease. Cancer cells may be associated with phenotypes such as uncontrolled proliferation, loss of specialized function, immortality, marked metastatic ability, markedly increased anti-apoptotic activity, rapid growth rate and rapid proliferation rate, as well as certain characteristic morphologies and cellular markers.

[0105] Under certain circumstances, cancer cells may take the form of a tumor, and these cells may exist locally within an animal (e.g., a solid tumor), or cancer cells may circulate in the bloodstream as independent cells, such as leukemia cells (non-solid tumors), or be dispersed throughout the body (e.g., metastasis). It will be understood that the term “cancer” as used herein encompasses any type of cancer, at any stage and in any form.

[0106] Types of malignant diseases to which the diagnostic or therapeutic methods of some embodiments of the present invention can be applied include benign tumors, warts, polyps, precancerous conditions, and malignant tumors / cancers.

[0107] Specific examples of cancerous diseases that can be treated using the methods of the present invention include, but are not limited to, carcinomas, lymphomas, blastomas, sarcomas, and leukemias. More specific examples of such cancers include squamous cell carcinoma, soft tissue sarcoma, Kaposi's sarcoma, melanoma, lung cancer (including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer or stomach cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, rectal cancer, endometrial cancer or uterine cancer, carcinoid carcinoma, salivary gland cancer, kidney cancer or renal cancer, liver cancer, prostate cancer, vulvar cancer, thyroid cancer, liver carcinoma, mesothelioma, multiple myeloma, post-transplant lymphoproliferative disorder (PTLD), and various types of head and neck cancers (e.g., brain tumors). The cancerous conditions to which the treatment of the present invention is applicable include metastatic cancer.

[0108] According to one embodiment, the malignant disease is a hematological malignancy. Exemplary hematological malignancies include leukemia [e.g., acute lymphoblastic leukemia, acute lymphoblastic leukemia, pre-B cell acute lymphoblastic leukemia, acute lymphoblastic T cell leukemia, acute megakaryoblastic leukemia, monocytic leukemia, acute myelogenous leukemia, acute myeloid leukemia, acute myeloid leukemia with eosinophilia, B cell leukemia, basophilic leukemia, chronic bone marrow leukemia]. Myelin leukemia, chronic leukemia, B-cell leukemia, eosinophilic leukemia, Friend leukemia, granulocytic or myeloid leukemia, hairy cell leukemia, lymphocytic leukemia, megakaryoblastic leukemia, monocytic leukemia, monocyte-macrophage leukemia, myeloblastic leukemia, myeloid leukemia, myelomonocytic leukemia, plasmacytic leukemia, pre-B-cell leukemia, promyelocytic leukemia, subacute leukemia, T-cell leukemia, lymphoid neoplasm [Patient predisposition to myeloid malignancies, acute non-lymphocytic leukemia, T-cell acute lymphoblastic leukemia (T-ALL), and B-cell chronic lymphocytic leukemia (B-CLL)], as well as lymphomas [e.g., Hodgkin's disease, non-Hodgkin lymphoma, Burkitt lymphoma, cutaneous T-cell lymphoma, histiocytic lymphoma, lymphoblastic lymphoma, T-cell lymphoma, thymic lymphoma, B-cell lymphoma (low-grade)]. This includes, but is not limited to, follicular lymphoma (including follicular lymphoma), small lymphocytic (SL) NHL, intermediate-grade follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-incisional nuclear cell NHL, bulky lesion NHL, mantle cell lymphoma, AIDS-associated lymphoma, and Waldenström macroglobulinemia.

[0109] In specific embodiments, malignant diseases include leukemia, lymphoma, myeloma, melanoma, sarcoma, neuroblastoma, colon cancer, colorectal cancer, breast cancer, ovarian cancer, esophageal cancer, synovial cell carcinoma, liver cancer, and pancreatic cancer.

[0110] According to one embodiment, the subject has a non-malignant disease.

[0111] According to one embodiment, non-malignant diseases include organ dysfunction or organ failure (e.g., kidney or liver disease associated with cytopenia), hematological non-malignant diseases (e.g., cytopenia, anemia, sickle cell anemia, hemophilia), transplant-related diseases (e.g., graft rejection), immunodeficiency, severe combined immunodeficiency syndrome (SCID), genetic disorders, metabolic disorders, infectious diseases, inflammatory diseases (e.g., systemic inflammatory diseases such as those associated with cytopenia), autoimmune diseases, allergic diseases, trauma, and injury.

[0112] Inflammatory diseases include, but are not limited to, chronic and acute inflammatory diseases.

[0113] Inflammatory diseases associated with hypersensitivity Examples of hypersensitivity include, but are not limited to, type I hypersensitivity, type II hypersensitivity, type III hypersensitivity, type IV hypersensitivity, immediate-type hypersensitivity, antibody-mediated hypersensitivity, immune complex-mediated hypersensitivity, T lymphocyte-mediated hypersensitivity, and DTH.

[0114] Type I or immediate-type hypersensitivity reactions such as asthma.

[0115] Type II allergy, RIMAC-sama disease, RIMAC-sama autoimmune disease, joint RIMAC (Krenn V. et al., Histol Histopathol 2000 Jul;15 (3):791), spondylitis, ankylosing spondylitis (Jan Voswinkel et al., Arthritis Res 2001; 3 (3): 189), systemic disease, systemic autoimmune disease, systemic ERITEMA TORDS (Erikson J. et al., Immunol Res 1998;17 (1-2):49), sclerosis, systemic sclerosis (Renaudineau Y. et al., Clin Diagn Lab Immunol. 1999 Mar;6 (2):156); Chan OT. et al., Immunol Rev 1999 Jun;169:107), glandular diseases, glandular autoimmune diseases, thyroid autoimmune diseases, diabetes, type I diabetes (Zimmet P. Diabetes Res Clin Pract 1996 Oct;34 Suppl:S125), thyroid diseases, autoimmune thyroid diseases, gravidarum (Orgiazzi J. Endocrinol Metab Clin North Am 2000 Jun;29 (2):339), thyroiditis, spontaneous autoimmune thyroiditis (Braley-Mullen H. and Yu S, J Immunol 2000 Dec 15;165 (12):7262), Hashimoto's thyroiditis (Toyoda N. et al., Nippon Rinsho 1999 Aug;57 (8):1810), myxedema, idiopathic myxedema (Mitsuma T. Nippon Rinsho). 1999 Aug;57 (8):1759); Autoimmune reproductive disorders, oviductal disorders, oviductal autoimmunity (Garza KM. et al., J Reprod Immunol 1998 Feb;37 (2):87), autoimmune anti-sperm infertility (Diekman AB. et al., Am J Reprod Immunol. 2000 Mar;43 (3):134), recurrent fetal loss (Tincani A. et al.)., Lupus 1998;7 Suppl 2:S107-9), neurodegenerative diseases, neurological diseases, neurological autoimmune diseases, multiple sclerosis (Cross AH. et al., J Neuroimmunol 2001 Jan 1;112 (1-2):1), Alzheimer's disease (Oron L. et al., J Neural Transm Suppl. 1997;49:77), myasthenia gravis (Infante AJ. And Kraig E, Int Rev Immunol 1999;18 (1-2):83), motor neuropathy (Kornberg AJ. J Clin Neurosci. 2000 May;7 (3):191), Guillain-Barre syndrome, neuropathy, and autoimmune neuropathy (Kusunoki S. Am J Med Sci. 2000 Apr;319 (4):234), myasthenic disorder, Lambert-Eaton myasthenic syndrome (Takamori M. Am J Med Sci. 2000 Apr;319 (4):204), paraneoplastic neurological disorders, cerebellar atrophy, paraneoplastic cerebellar atrophy, non-paraneoplastic Stiffman syndrome, cerebellar atrophy, progressive cerebellar atrophy, encephalitis, Rasmussen encephalitis, amyotrophic lateral sclerosis, Sydenham's chorea, Gilles de la Tourette syndrome, polyglandular endocrine disorder, autoimmune polyglandular endocrine disorder (Antoine JC. and Honnorat J. Rev Neurol (Paris) 2000 Jan;156 (1):23); neuropathy, immune-abnormal neuropathy (Nobile-Orazio E. et al., Electroencephalogr Clin Neurophysiol Suppl 1999;50:419); Neuromyotonia, acquired neuromyotonia, congenital multiple arthroplasty (Vincent A. et al., Ann NY Acad Sci. 1998 May 13;841:482), cardiovascular disease, cardiovascular autoimmune disease, atherosclerosis (Matsuura E. et al., Lupus. 1998;7 Suppl 2:S135), myocardial infarction (Vaarala O. Lupus. 1998;7 Suppl 2:S132), thrombosis (Tincani A. et al., Lupus 1998;7 Suppl 2:S107-9), granulomatous disease, vegenerous granulomatous disease, arteritis, high-an arteritis, Kawasaki syndrome (Praprotnik S. et al., Wien Klin Wochenschr 2000 Aug 25;112 (15-16):660); anti-factor VIII autoimmune disease (Lacroix-Desmazes S. et al., Semin Thromb Hemost.2000;26 (2):157); Vasculitis, catastrophic small vessel vasculitis, microscopic multivasculitis, Charg-Strauss syndrome, glomerulonephritis, microimmune nest-like catastrophic glomerulonephritis, hemispheric glomerulonephritis (Noel LH. Ann Med Interne (Paris). 2000 May;151 (3):178); Anti-Rin lipid antibody syndrome (Flamholz R. et al., J Clin Apheresis 1999;14 (4):171); Heart failure, heart failure β-adrenalin receptor antibody (Wallukat G. et al., Am J Cardiol. 1999 Jun 17;83 (12A):75H), thrombocytopenic purpura (Moccia F. Ann Ital Med Int. 1999 Apr-Jun;14 (2):114); hemolytic anemia, autoimmune hemolytic anemia (Efremov DG. et al., Leuk Lymphoma 1998 Jan;28 (3-4):285), gastrointestinal disorders, autoimmune disorders of the gastrointestinal tract, intestinal disorders, chronic inflammatory bowel disease (Garcia Herola A. et al., Gastroenterol Hepatol. 2000 Jan;23 (1):16), ceric disease (Landau YE. and Shoenfeld Y. Harefuah 2000 Jan 16;138) (2):122), muscle system autoimmune disorders, muscle inflammation, autoimmune muscle inflammation, schigren syndrome (Feist E. et al., Int Arch Allergy Immunol 2000 Sep;123 (1):92); smooth muscle autoimmune disorders (Zauli D. et al., Biomed Pharmacother 1999 Jun;53 (5-6):234), liver diseases, liver autoimmune disorders, autoimmune hepatitis (Manns MP).This includes, but is not limited to, primary biliary cirrhosis (J Hepatol 2000 Aug;33 (2):326) and primary biliary cirrhosis (Strassburg CP. et al., Eur J Gastroenterol Hepatol. 1999 Jun;11 (6):595).

[0116] Type IV hypersensitivity or T-cell-mediated hypersensitivity includes rheumatic-like diseases, rheumatoid arthritis (Tisch R, McDevitt HO. Proc Natl Acad Sci U S A 1994 Jan 18;91 (2):437), systemic diseases, systemic autoimmune diseases, systemic lupus erythematosus (Datta SK., Lupus 1998;7 (9):591), glandular diseases, glandular autoimmune diseases, pancreatic diseases, pancreatic autoimmune diseases, type 1 diabetes (Castano L. and Eisenbarth GS. Ann. Rev. Immunol. 8:647); thyroid diseases, autoimmune thyroid diseases, Graves' disease (Sakata S. et al., Mol Cell Endocrinol 1993 Mar;92 (1):77); ovarian diseases (Garza KM. et al., J Reprod Immunol 1998 Feb;37 (2):87), prostatitis, autoimmune prostatitis (Alexander RB. et al., Urology 1997 Dec;50 (6):893), polyglandular syndromes, autoimmune polyglandular syndromes, type I autoimmune polyglandular syndrome (Hara T. et al., Blood. 1991 Mar 1;77 (5):1127), neurological diseases, autoimmune neurological diseases, multiple sclerosis, neuritis, optic neuritis (Soderstrom M. et al., J Neurol Neurosurg Psychiatry 1994 May;57 (5):544), myasthenia gravis (Oshima M. et al., Eur J Immunol 1990 Dec;20 (12):2563), stiff-man syndrome (Hiemstra HS. et al., Proc Natl Acad Sci U S A 2001 Mar 27;98 (7):3988), cardiovascular diseases, cardiac autoimmunity in Chagas disease (Cunha-Neto E. et al., J Clin Invest 1996 Oct 15;98 (8):1709), autoimmune thrombocytopenic purpura (Semple JW. et al., Blood 1996 May 15;87 (10):4245), anti-helper T lymphocyte autoimmunity (Caporossi AP. et al., Viral Immunol 1998;11 (1):9), hemolytic anemia (Sallah S. et al., Ann Hematol 1997 Mar;74 (3):139), liver disease, liver autoimmune disease, hepatitis, chronic active hepatitis (Franco A. et al., Clin Immunol Immunopathol 1990 Mar;54) (3):382), biliary cirrhosis, primary biliary cirrhosis (Jones DE. Clin Sci (Colch) 1996 Nov;91 (5):551), renal disease, renal autoimmune disease, nephritis, interstitial nephritis (Kelly CJ. J Am Soc Nephrol 1990 Aug;1 (2):140), connective tissue disease, ear disease, autoimmune connective tissue disease, autoimmune ear disease (Yoo TJ. et al., Cell Immunol 1994 Aug;157 (1):249), inner ear disorders (Gloddek B. et al., Ann NY Acad Sci 1997 Dec 29;830:266), skin diseases, cutaneous diseases, dermal diseases, bullous skin diseases, pemphigus vulgaris, bullous pemphigoid, and pemphigus foliaceus, but are not limited to these.

[0117] Examples of delayed-type hypersensitivity include, but are not limited to, contact dermatitis and drug rash.

[0118] Examples of T lymphocyte types that mediate hypersensitivity include, but are not limited to, helper T lymphocytes and cytotoxic T lymphocytes.

[0119] Examples of helper T lymphocyte-mediated hypersensitivity include, but are not limited to, Th1 lymphocyte-mediated hypersensitivity and Th2 lymphocyte-mediated hypersensitivity.

[0120] autoimmune disease This includes, but is not limited to, cardiovascular diseases, rheumatic diseases, glandular diseases, gastrointestinal diseases, skin diseases, liver diseases, neurological diseases, muscle diseases, kidney diseases, reproductive disorders, connective tissue diseases, and systemic diseases.

[0121] Examples of autoimmune cardiovascular diseases include atherosclerosis (Matsuura E. et al., Lupus. 1998;7 Suppl 2:S135), myocardial infarction (Vaarala O. Lupus. 1998;7 Suppl 2:S132), and thrombosis (Tincani A. et al., Lupus 1998;7 Suppl 2:S107-9), Wegener's granulomatosis, Takayasu arteritis, Kawasaki syndrome (Praprotnik S. et al., Wien Klin Wochenschr 2000 Aug 25;112 (15-16):660), anti-factor VIII autoimmune disease (Lacroix-Desmazes S. et al., Semin Thromb Hemost.2000;26 (2):157); Necrotizing small vessel vasculitis, microscopic polyangiitis, Churg-Strauss syndrome, microimmune focal necrotizing glomerulonephritis and crescentic glomerulonephritis (Noel LH. Ann Med Interne (Paris). 2000 May;151 (3):178), antiphospholipid antibody syndrome (Flamholz R. et al., J Clin Apheresis) 1999;14 (4):171), antibody-induced heart failure (Wallukat G. et al., Am J Cardiol. 1999 Jun 17;83 (12A):75H), thrombocytopenic purpura (Moccia F. Ann Ital Med Int. 1999 Apr-Jun;14 (2):114;Semple JW. et al., Blood 1996 May 15;87 This includes, but is not limited to, autoimmune hemolytic anemia (Efremov DG. et al., Leuk Lymphoma 1998 Jan;28 (3-4):285;Sallah S. et al., Ann Hematol 1997 Mar;74 (3):139), cardiac autoimmunity in Chagas disease (Cunha-Neto E. et al., J Clin Invest 1996 Oct 15;98 (8):1709), and anti-helper T lymphocyte autoimmunity (Caporossi AP. et al., Viral Immunol 1998;11 (1):9).

[0122] Examples of autoimmune rheumatoid-like diseases include, but are not limited to, rheumatoid arthritis (Krenn V. et al., Histol Histopathol 2000 Jul;15 (3):791; Tisch R, McDevitt HO. Proc Natl Acad Sci units SA 1994 Jan 18;91 (2):437) and ankylosing spondylitis (Jan Voswinkel et al., Arthritis Res 2001; 3 (3): 189).

[0123] Examples of autoimmune gonadal disorders include, but are not limited to, pancreatic diseases, type 1 diabetes, thyroid diseases, Graves' disease, thyroiditis, spontaneous autoimmune thyroiditis, Hashimoto's thyroiditis, idiopathic myxedema, ovarian autoimmunity, autoimmune antispermia, autoimmune prostatitis, and type 1 autoimmune polyglandular syndrome. Diseases include autoimmune diseases of the pancreas, type 1 diabetes (Castano L. and Eisenbarth GS. Ann. Rev. Immunol. 8:647;Zimmet P. Diabetes Res Clin Pract 1996 Oct;34 Suppl:S125), autoimmune thyroid disease, and Graves' disease (Orgiazzi J. Endocrinol Metab Clin North Am 2000 Jun;29 (2):339;Sakata S. et al., Mol Cell Endocrinol 1993 Mar;92 (1):77), spontaneous autoimmune thyroiditis (Braley-Mullen H. and Yu S, J Immunol 2000 Dec 15;165 (12):7262), Hashimoto's thyroiditis (Toyoda N. et al., Nippon Rinsho 1999 Aug;57) (8):1810), idiopathic myxedema (Mitsuma This includes, but is not limited to, T. Nippon Rinsho. 1999 Aug;57 (8):1759, ovarian autoimmunity (Garza KM. et al., J Reprod Immunol 1998 Feb;37 (2):87), autoimmune antisperm infertility (Diekman AB. et al., Am J Reprod Immunol. 2000 Mar;43 (3):134), autoimmune prostatitis (Alexander RB. et al., Urology 1997 Dec;50 (6):893), and type I autoimmune polyglandular syndrome (Hara T. et al., Blood. 1991 Mar 1;77 (5):1127).

[0124] Examples of autoimmune gastrointestinal diseases include, but are not limited to, chronic inflammatory bowel disease (Garcia Herola A. et al., Gastroenterol Hepatol. 2000 Jan;23 (1):16), celiac disease (Landau YE. and Shoenfeld Y. Harefuah 2000 Jan 16;138 (2):122), colitis, ileitis, and Crohn's disease.

[0125] Examples of autoimmune skin diseases include, but are not limited to, autoimmune bullous skin diseases (such as pemphigus vulgaris, bullous pemphigoid, and pemphigus foliaceus).

[0126] Examples of autoimmune liver diseases include, but are not limited to, hepatitis, autoimmune chronic active hepatitis (Franco A. et al., Clin Immunol Immunopathol 1990 Mar;54 (3):382), primary biliary cirrhosis (Jones DE. Clin Sci (Colch) 1996 Nov;91 (5):551; Strassburg CP. et al., Eur J Gastroenterol Hepatol. 1999 Jun;11 (6):595), and autoimmune hepatitis (Manns MP. J Hepatol 2000 Aug;33 (2):326).

[0127] Examples of autoimmune neurological disorders include multiple sclerosis (Cross AH. et al., J Neuroimmunol 2001 Jan 1;112 (1-2):1), Alzheimer's disease (Oron L. et al., J Neural Transm Suppl. 1997;49:77), myasthenia gravis (Infante AJ. And Kraig E, Int Rev Immunol 1999;18 (1-2):83; Oshima M. et al., Eur J Immunol 1990 Dec;20 (12):2563), neuropathy, motor neuropathy (Kornberg AJ. J Clin Neurosci. 2000 May;7 (3):191), Guillain-Barré syndrome, and autoimmune neuropathy (Kusunoki S. Am J Med Sci. 2000 Apr;319) (4):234), myasthenia gravis, Lambert-Eaton myasthenic syndrome (Takamori M. Am J Med Sci. 2000 Apr;319 (4):204); paraneoplastic neurological disorders, cerebellar atrophy, paraneoplastic cerebellar atrophy, and Stiffman syndrome (Hiemstra HS. et al., Proc Natl Acad Sci units SA 2001 Mar 27;98 (7):3988); non-paraneoplastic Stiffman syndrome, progressive cerebellar atrophy, encephalitis, Rasmussen encephalitis, amyotrophic lateral sclerosis, Sydenham chorea, Gilles de la Tourette syndrome, and autoimmune polyglandular endocrine disorders (Antoine JC. and Honnorat J. Rev Neurol (Paris) 2000 Jan;156 (1):23); immunological neuropathy (Nobile-Orazio E. et al., Electroencephalogr Clin Neurophysiol Suppl 1999;50:419); Acquired neuromyotonia, congenital multiple arthrocontractures (Vincent A. et al., Ann NY Acad Sci. 1998 May 13;841:482), neuritis, optic neuritis (Soderstrom M. et al.)This includes, but is not limited to, neurodegenerative diseases (J Neurol Neurosurg Psychiatry 1994 May;57 (5):544) and neurodegenerative diseases.

[0128] Examples of autoimmune muscle diseases include, but are not limited to, myositis, autoimmune myositis, and primary Sjögren's syndrome (Feist E. et al., Int Arch Allergy Immunol 2000 Sep;123 (1):92), as well as smooth muscle autoimmune diseases (Zauli D. et al., Biomed Pharmacother 1999 Jun;53 (5-6):234).

[0129] Examples of autoimmune kidney diseases include, but are not limited to, nephritis and autoimmune interstitial nephritis (Kelly CJ. J Am Soc Nephrol 1990 Aug;1 (2):140).

[0130] Examples of reproductive autoimmune diseases include, but are not limited to, recurrent fetal loss (Tincani A. et al., Lupus 1998;7 Suppl 2:S107-9).

[0131] Examples of autoimmune connective tissue diseases include, but are not limited to, ear diseases, autoimmune ear diseases (Yoo TJ. et al., Cell Immunol 1994 Aug;157 (1):249), and autoimmune diseases of the inner ear (Gloddek B. et al., Ann NY Acad Sci 1997 Dec 29;830:266).

[0132] Examples of autoimmune systemic diseases include, but are not limited to, systemic lupus erythematosus (Erikson J. et al., Immunol Res 1998;17 (1-2):49) and systemic sclerosis (Renaudineau Y. et al., Clin Diagn Lab Immunol. 1999 Mar;6 (2):156); Chan OT. et al., Immunol Rev 1999 Jun;169:107).

[0133] infectious disease Examples of infectious diseases include, but are not limited to, chronic infectious diseases, subacute infectious diseases, acute infectious diseases, viral diseases, bacterial diseases, protozoal diseases, parasitic diseases, fungal diseases, mycoplasma diseases, and prion diseases.

[0134] Specific types of viral pathogens that cause infectious diseases treatable according to the teachings of the present invention include, but are not limited to, retroviruses, circoviruses, parvoviruses, papovaviruses, adenoviruses, herpesviruses, iridoviruses, poxviruses, hepadnaviruses, picornaviruses, caliciviruses, togaviruses, flaviviruses, reoviruses, orthomyxoviruses, paramyxoviruses, rhabdoviruses, bunyaviruses, coronaviruses, arenaviruses, and filoviruses.

[0135] Specific examples of viral infections that can be treated in accordance with the teachings of the present invention include, but are not limited to, viral infections caused by human immunodeficiency virus (HIV)-induced acquired immunodeficiency syndrome (AIDS), influenza, rhinovirus infection, viral meningitis, Epstein-Barr virus (EBV) infection, hepatitis A virus infection, hepatitis B virus infection, or hepatitis C virus infection, measles, papillomavirus infection / verruca, cytomegalovirus (CMV) infection, herpes simplex virus infection, yellow fever, Ebola virus infection, rabies, adenovirus (Adv), common cold virus, influenza virus, Japanese encephalitis, polio, respiratory syncytium, rubella, smallpox, varicella-zoster, rotavirus, West Nile virus, and Zika virus.

[0136] Specific examples of bacterial infections that can be treated according to the teachings of the present invention include, but are not limited to, bacterial infections caused by anthrax, Gram-negative bacilli, Chlamydia, diphtheria, Haemophilus influenzae, Helicobacter pylori, malaria, Mycobacterium tuberculosis, pertussis toxin, pneumococcus, rickettsia, staphylococcus, streptococcus, and tetanus.

[0137] Specific examples of superbug infections (e.g., multidrug-resistant bacteria) that can be treated according to the teachings of the present invention include, but are not limited to, superbug infections caused by Enterococcus faecium, Clostridium difficile, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacteriaceae (including Escherichia coli, Klebsiella pneumoniae, and species of the genus Enterobacter).

[0138] Specific examples of fungal infections that can be treated according to the teachings of the present invention include, but are not limited to, fungal infections caused by Candida, Coccidioides, Cryptococcus, Histoplasma, Leishmania, Plasmodium, Protozoa, Parasites, Schistosomiasis, Tinea, Toxoplasma, and Trypanosoma cruzi.

[0139] Graft rejection disorder In other embodiments, the disease is related to graft transplantation. Examples of diseases related to graft transplantation include, but are not limited to, graft rejection, chronic graft rejection, subacute graft rejection, hyperacute graft rejection, acute graft rejection, allograft rejection, xenograft rejection, and graft-versus-host disease (GVHD).

[0140] Allergic diseases Examples of allergic diseases include, but are not limited to, asthma, hives, urticaria, pollen allergies, dust mite allergies, poison venom allergies, cosmetic allergies, latex allergies, chemical allergies, drug allergies, insect bite allergies, animal scale allergies, stinging plant allergies, poison ivy allergies, and food allergies.

[0141] Non-malignant blood diseases Examples of non-malignant hematological disorders include, but are not limited to, cytopenia (e.g., anemia, leukopenia, neutropenia, thrombocytopenia, granulocytopenia, pancytopenia), drug-induced cytopenia, toxin-induced cytopenia, radiation-induced cytopenia, or cytopenia associated with conventional bone marrow transplantation, bone marrow disorders, deep vein thrombosis / pulmonary embolism, Diamond-Blackfan anemia, hemochromatosis, hemophilia, immunohematological disorders, iron metabolism disorders, sickle cell anemia, thalassemia, osteopetrosis, von Willebrand disease, and Gaucher disease.

[0142] In addition, the method of mobilizing hematopoietic stem cells can be used for patients who are "difficult to mobilize" (for example, due to lack of sensitivity to growth factors).

[0143] The hematopoietic stem cells of the present invention can be transplanted into a recipient using any method known in the art for cell transplantation, for example, but not limited to, cell injection (e.g., IV) or via an intraperitoneal route.

[0144] As used herein, the term “approximately” refers to a range of ±10%.

[0145] The terms "comprises," "comprising," "includes," "including," and "having," as well as their conjugations, all mean "to include, but not limited to."

[0146] The term "consists of" means "includes and is limited to."

[0147] The term “essentially derived from” means that the composition, method, or structure may include additional components, processes, and / or parts, but only if the additional components, processes, and / or parts do not substantially alter the basic and novel features of the claimed composition, method, or structure.

[0148] As used herein, the singular forms "a," "an," and "the" refer to multiple objects unless otherwise explicitly indicated by the context. For example, the terms "a compound" or "at least one compound" may refer to multiple compounds (including mixtures thereof).

[0149] Throughout this application, various embodiments of the invention may be presented in range form. It should be understood that the range form is merely for convenience and brevity and should not be interpreted as an irrevocable limitation on the scope of the invention. Therefore, a range description should be considered to specifically disclose all possible subranges, as well as the individual numerical values ​​within that range. For example, a range description such as 1-6 should be considered to specifically disclose not only subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, as well as the individual numerical values ​​within that range, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the width of the range.

[0150] Wherever a numerical range is indicated herein, it is understood to include any of the listed numbers (fractions or integers) within that range. The phrases “range between” the first number and the second number, and “range from” the first number to the second number, are used interchangeably herein and mean including the first and second numbers, as well as all fractions and integers between them.

[0151] As used herein, the term “method” means a set of forms, means, techniques, and procedures for achieving a given task, which include, but are not limited to, forms, means, techniques, and procedures known to practitioners in the fields of chemistry, pharmacology, biology, biochemistry, and medicine, or forms, means, techniques, and procedures readily developed by such practitioners from known forms, means, techniques, and procedures.

[0152] As used herein, the term “treat” includes inhibiting, substantially inhibiting, delaying, or reversing the progression of a condition, substantially alleviating the clinical or aesthetic symptoms of a condition, or substantially preventing the appearance of the clinical or aesthetic symptoms of a condition.

[0153] For clarity, certain features of the Invention described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, various features of the Invention described in the context of a single embodiment for brevity may also be provided separately, in any preferred partial combination, or as suitably provided in any other embodiment described of the Invention. Certain features described in the context of various embodiments should not be considered essential features of those embodiments unless the embodiments would be non-functional without those elements.

[0154] While the present invention has been described in conjunction with its specific embodiments, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Therefore, it is intended that all such alternatives, modifications, and variations that fall within the spirit and broad scope of the appended claims are encompassed.

[0155] All publications, patents, and patent applications referenced herein are incorporated herein by reference in their entirety, as if each individual publication, patent, or patent application were specifically and individually stated to be incorporated herein by reference. In addition, any citation or specification of references in this application should not be construed as an acceptance that such references are available as prior art of the present invention. Section headings, insofar as they are used, should not necessarily be construed as limiting. In addition, any priority documents of this application are incorporated herein by reference in their entirety.

Claims

1. A method for recruiting hematopoietic progenitor cells from the target bone marrow to the peripheral blood, (a) the step of administering an effective amount of natalizumab to the subject, and (b) A step of administering an effective amount of CXCR4 inhibitor to the subject, which is performed at least 48 hours after step (a). Methods that include...

2. A method for obtaining a therapeutically effective amount of hematopoietic progenitor cells from a subject, (a) A step of administering an effective amount of natalizumab to the subject, (b) a step of administering an effective amount of CXCR4 inhibitor to the subject, which is performed at least 48 hours after step (a), and (c) A step of collecting the hematopoietic progenitor cells by apheresis. Methods that include...

3. An effective combination of natalizumab and a CXCR4 inhibitor for use in recruiting hematopoietic progenitor cells in a subject, wherein the CXCR4 inhibitor is administered to the subject at least 48 hours after administration of natalizumab.

4. An effective combination of natalizumab and a CXCR4 inhibitor for use in a method for collecting hematopoietic progenitor cells by apheresis, wherein the method comprises mobilizing the hematopoietic progenitor cells, the hematopoietic progenitor cells being collected by subsequent apheresis, and the CXCR4 inhibitor being administered to the subject at least 48 hours after the administration of natalizumab.

5. The method or combination for use according to any one of claims 1 to 3, wherein the subject is a human subject.

6. The method or combination for use according to any one of claims 1 to 3, wherein the subject is a donor of hematopoietic progenitor cells.

7. The method or combination for use according to any one of claims 1 to 3, wherein the subject has been diagnosed with cancer.

8. The method according to any one of claims 1 to 3, or a combination for use, wherein the hematopoietic progenitor cells are for autologous transplantation.

9. The method according to any one of claims 1 to 3, or a combination for use, wherein the hematopoietic progenitor cells are for allogeneic transplantation.

10. The method or combination for use according to any one of claims 1 to 9, wherein the effective amount comprises repeated cycles of administration of the natalizumab and the CXCR4 inhibitor.

11. The method or combination for use according to any one of claims 1 to 10, wherein the aforementioned at least 48 hours include a maximum of 96 hours.

12. A method or combination for use according to any one of claims 1 to 10, wherein the aforementioned at least 48 hours include a maximum of 72 hours.

13. The method according to any one of claims 1 to 12, or a combination for use, wherein the CXCR4 inhibitor is a peptide, a small molecule, an antibody, a nucleic acid, or a combination thereof.

14. The method or combination for use according to any one of claims 1 to 12, wherein the CXCR4 inhibitor is the peptide described in SEQ ID NO:

1.

15. The method or combination for use according to any one of claims 1 to 12, wherein the CXCR4 inhibitor is a small molecule.

16. The method or combination for use according to any one of claims 1 to 12, wherein the CXCR4 inhibitor is AMD3100.

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