GPCR Inhibitors and Their Uses

Abstract

The present invention relates to a method and composition for mobilizing cells from a subject by blocking CXCR4, beta-adrenergic receptors, GPCRs, or any combination thereof. In some embodiments, the cells are hematopoietic stem cells. In some embodiments, the method further comprises the administration of G-CSF. The present invention also relates to a treatment method, a method for qualifying a subject for treatment, and a method for preparing a subject for treatment by blocking CXCR4, beta-adrenergic receptors, GPCRs, or any combination thereof. In some embodiments, the cells are hematopoietic stem cells. In some embodiments, the treatment is for cancer. In some embodiments, CXCR4 is GPC-100. In some embodiments, the method further comprises the administration of G-CSF.

Description

Technology Field

[0001] This technology generally relates to GPCR inhibitors and their use in the treatment of diseases. Background Technology

[0002] Multiple myeloma (MM) is a major hematologic malignancy, with approximately 34,920 cases annually in the United States and 588,161 cases worldwide [1]. Autologous Stem Cell Transplant (ASCT) is essential for the overall management of eligible MM patients and has improved anticancer response and survival rates compared to traditional chemotherapy [2-5]. The success of ASCT depends on harvesting a sufficient number of hematopoietic stem cells (HSCs), which is primarily achieved by mobilizing HSCs from the bone marrow (BM) into peripheral blood (PB) [6, 7]. In humans, HSCs are phenotypically characterized by the expression of CD34. For HSC harvesting, at least 2 × 10⁶ 6 CD34 + Cells / kg is essential, and the optimal number for improved engraftment and survival is >5-6×10 6 CD34 +It is cells / kg [8, 9]. Granulocyte-colony-stimulating factor (G-CSF) is the clinical standard treatment for HSC mobilization

[10] . However, G-CSF fails to mobilize an optimal number of HSCs in at least 40–50% of MM patients [10, 11]. Some patients are treated with a combination of G-CSF and the small molecule CXCR4 antagonist AMD3100 (Plerixafor or Mozobil)

[10] . Despite this combination therapy, 15–35% of MM patients fail to mobilize a sufficient number of cells [10, 12]. In a recent Phase 3 clinical study, the combination of G-CSF and the CXCR4 peptide inhibitor BL8040 (motixafortide) resulted in significantly more CD34 cells compared to the combination of G-CSF and placebo. + Cells were mobilized

[13] . While this is a promising result, this study did not include patients receiving the recently approved daratumumab

[14] , and the accumulated data suggest that daratumumab may have a much more negative impact on HSC mobilization than the traditional treatment lenalidomide

[15] . Failure of mobilization can lead to the potential loss of ASCT as a treatment option and significant toxicity resulting from repeated mobilization attempts. Additionally, G-CSF is contraindicated for stem cell collection in conditions such as sickle cell disease

[16] and exacerbates autoimmune diseases

[17] . The problem to be solved

[0003] Therefore, there is a need to identify new strategies that not only address the problem of poor mobilization in MM patients but also provide non-G-CSF options for appropriate mobilization in MM and other disease indications [18, 19]. Brief explanation of the drawing

[0004] Fig. 1a-1bThis shows a molecular-level comparison of the CXCR4 inhibitors GPC-100 and AMD3100. Fig. 1a Chemical structures of AMD3100 and GPC-100. Fig. 1b ) Schrödinger(Schr Overlay of the proposed binding modes of GPC-100 and AMD3100 obtained from molecular docking using Dinger's induced-fit algorithm and an inert structural template of CXCR4. The CXCR4 transmembrane helix is ​​indicated by a gray ribbon and annotated. The 2D ligand interaction diagram shows amino acid residues as colored triangular peaks. Outward-facing peaks represent the backbones of the residues facing each ligand. Peaks facing the ligands represent the side chains of the residues facing the ligands. Fig. 2 Figure 1 shows dose-response curves demonstrating the competitive inhibition of CXCL12 binding to CXCR4 by GPC-100 and AMD3100. Ki for each compound was determined by plotting the normalized HTRF ratio (% CXCL12 binding) versus compound concentration and using the nonlinear regression competitive binding "one site-fit Ki equation." Data are expressed as mean ± SEM. Figs. 3a-3d This demonstrates the pharmacological inhibition of the CXCR4 / CXCL12 axis by GPC-100 and AMD3100. Fig. 3a Inhibition of CXCL12(20 nM)-induced calcium flux by GPC-100 and AMD3100 in CXCR4-transduced MDA-MB-231 cells. Fig. 3b Inhibition of CXCL12-induced β-arrestin recruitment to CXCR4 by GPC-100 and AMD3100 using Presto-Tango assay in HTLA cells overexpressing CXCR4-tango and β2AR. Figs. 3c-3dInhibition of CXCL12-induced migration in human cells by GPC-100 and AMD3100. Fig. 3c Inhibition profile in U937 cells. Fig. 3d Inhibition profile in MM.1S cells. Data are expressed as mean ± SEM. Fig. 4 This shows the expression of CXCR4 in U937 cells. U937 cells were pretreated with a polyclonal antibody that inhibits the Fc receptor binding, and surface expression of CXCR4 was detected by flow cytometry using an Alexa Fluor 568-conjugated goat anti-human IgG antibody followed by an isotype control or an anti-CXCR4 antibody (ulocuplumab). Figs. 5a-5b ... demonstrates CXCL12-induced migration. 5 x 10 cells in serum-free medium containing 0.5% BSA 6 Cells were suspended at a concentration of 100 μl / ml, and 100 μl of the cell suspension was added to a transwell insert, wherein the transwell contains increasing concentrations of CXCL12. Fig. 5a Migration profile in MM.1S cells. Fig. 5b Migration profile in U937 cells. Figs. 6a-6b It shows the expression of CXCR4 and β2AR in Namalwa cells. Flow cytometry was used to detect surface expression of endogenous CXCR4 and β2AR in Namalwa cells after staining with anti-CXCR4 or anti-β2AR antibodies. Fig. 6a ) Endogenous CXCR4 profile visualized using PE-conjugated anti-human CXCR4 antibody (1D9). ( Fig. 6b Endogenous β2AR visualized using Alexa-Fluor 488-conjugated anti-human β2AR(R11E1) antibody. Figs. 7a-7b... shows the co-localization of CXCR4 and β2AR in cancer cells. PLA showing the proximity between CXCR4 and β2AR in Namalwa cells. ( Fig. 7a ) Co-localization profiles in Namalva progenitor cells and Namalva CXCR4 knockout cells. Fig. 7b Co-localization profiles in MDA-MB-231 blast cells and MDA-MB-231 ADRB2 knockout cells. Data are expressed as mean ± SEM with statistical significance. Figs. 8a-8d It shows the validation of Namalwa-CXCR4 knockout and MDA-MB-231-ADRB2 knockout cells. Cell surface expression of CXCR4 in control and knockout-treated Namalwa cells was detected by flow cytometry using isotype control or PE-conjugated anti-human CXCR4 antibody (1D9) or Alexa-Fluor 488-conjugated anti-human β2AR (R11E1) antibody. Fig. 8a ) Flow cytometry profiles according to CRISPR / Cas9-sgControl control treatment. Fig. 8b ) Flow cytometry profiles according to CRISPR / Cas9-sgCXCR4 control treatment. Fig. 8c Flow cytometry showing cell surface expression of β2AR in progenitor cells and control cells. Fig. 8d Flow cytometry showing cell surface expression of β2AR in MDA-MB-231 cells treated with CRISPR / Cas9-sgADRB2. Figs. 9a-9c This demonstrates that co-activation of CXCR4 and β2AR leads to a synergistic increase in β-arrestin recruitment to CXCR4. HTLA cells overexpressing CXCR4-tango and β2AR were treated in two separate experiments. Fig. 9a ) β-arrestin recruitment profiles in the presence of a constant 1 μM epinephrine and various 0-200 nM CXCL12. Fig. 9b) β-arrestin recruitment profiles in the presence of a constant 100 nM CXCL12 and various 0-10 μM epinephrines. Co-treatment with epinephrine and CXCL12 induced a synergistic increase in β-arrestin recruitment. Fig. 9c ) Change in fold of β-arrestin recruitment to CXCR4 by co-treatment with 100 nM CXCL12, 400 nM epinephrine, agonist, 10 μM GPC-100, 10 μM propranolol, or antagonist. Data are expressed as mean ± SEM. Pro: Propranolol. Figs. 10a-10b It shows the expression of CXCR4 and β2AR in MDA-MB-231 cells. Anti-human CXCR4 (ulocuflumab) and Alexa Fluor 568-conjugated goat anti-human IgG antibody ( Fig. 10a ) and Alexa-Fluor 488-conjugated anti-human β2AR antibody (R11E1)( Fig. 10b Flow cytometry was used to detect surface expression of endogenous CXCR4 and β2AR in MDA-MB-231 cells after staining with ). Figs. 11a-11b This shows the effects of co-treatment with GPC-100 and propranolol on CXCL12 and epinephrine-induced crosstalk. Fig. 11a Calcium flux in MDA-MB-231 cells endogenously expressing CXCR4 and β2AR incubated with vehicle, 200 nM CXCL12, 10 μM epinephrine, or CXCL12 and epinephrine. Data represent a fold increase compared to the vehicle. Fig. 11b Inhibition of synergistic calcium increase by 10 μM GPC-100, 10 μM AMD3100, 10 μM propranolol, or co-treatment thereof in MDA-MB-231 cells treated with CXCL12 and epinephrine. Calcium flux percentages were normalized to CXCL12. Data are expressed as mean ± SEM with statistical significance. Pro: Propranolol. Figs. 12a-12b Figure 1 shows in vivo mobilization by GPC-100 or AMD3100. White blood cell (WBC) mobilization into peripheral blood (PB) after a single injection of GPC-100 (30 mg / kg, IV) or AMD3100 (5 mg / kg, SC). PB was collected 1 hour after drug administration. Fig. 12a Time course of mobilization by GPC-100 or AMD3100. PB was collected at various time points after drug administration. Blood was drawn twice from each mouse. Blood draws at 0.5, 1, and 2 hours were non-terminal, while those at 3, 4, and 5 hours were terminal. Figures 12a and 12b represent two independent studies. Data are expressed as mean ± SEM. Fig. 13 Figure 7 shows the dose-response of propranolol to the mobilization of GPC-100. Propranolol (0 to 40 mg / kg, IP) was administered for 7 days, and GPC-100 was co-administered on the 7th day. Data are expressed as mean ± SEM. Statistical significance *p<0.05, **p<0.01. Pro: Propranolol. Figs. 14a-14d It shows in vivo mobilization by GPC-100 and propranolol. Fig. 14a A dosing regimen indicating the co-administration of GPC-100 (30 mg / kg, IV) on day 7 following 7 days of propranolol (20 mg / kg, IP) pretreatment. AMD3100 (5 mg / kg, SC) was similarly co-administered with propranolol on day 7. Fig. 14b WBC mobilization by the combination of GPC-100 and propranolol, and AMD3100 and propranolol. Fig. 14c Analysis of representative LSK frequency in PB. Fig. 14dNumber of LSK cells mobilized in mice treated with PBS, GPC-100 alone, or in combination with propranolol. Data from two separate experiments. Pro: Propranolol. Data are expressed as mean ± SEM. Figs. 15a-15d Figure 1 shows the mobilization of lymphocytes and neutrophils by GPC-100 and AMD3100 alone and in combination with propranolol. Propranolol (20 mg / kg, IP) was administered for 7 days, and GPC-100 or AMD3100 was co-administered on the 7th day. Cell mobilization after GPC-100 treatment was measured depending on the co-administration of propranolol. Fig. 15a ) Lymphocyte mobilization profile. Fig. 15b ) Neutrophil mobilization profile. Cell mobilization was measured after AMD3100 treatment depending on the co-administration of propranolol. Fig. 15c ) Lymphocyte mobilization profile. Fig. 15d ) Neutrophil mobilization profile. Data are expressed as mean ± SEM. Statistical significance *p<0.05, **p<0.01 Figs. 16a-16h It demonstrates in vivo mobilization by a triple combination of GPC-100, propranolol, and G-CSF. Fig. 16a ) Administration regimen. G-CSF (0.1 mg / kg, SC) was administered twice daily (BID) for 5 days from Day 2 to Day 6, and propranolol (20 mg / kg, IP) was administered for 7 days. GPC-100 (30 mg / kg, IV) was co-administered with propranolol on Day 7. Mice in the triple combination group received all three treatments. For comparison with standard treatment, G-CSF was administered alone or in combination with AMD3100 (5 mg / kg SC), which was administered on Day 7, 12 hours after G-CSF administration. Fig. 16b ) WBC Mobilization. Fig. 16c ) LSK cells (Lin - Sca1 + ckit + Mobilization of ) Fig. 16d ) Colony-forming unit (CFU). Fig. 16e ) LSK cells (CD34 - Mobilization of ) Fig. 16f LSK cells (CD150) evaluated by flow cytometry + Mobilization of ). G: G-CSF. Propranolol administration to Lin lacking CD34 expression = Sca-1 + cKit + It improves GPC-100-induced recruitment of primitive cells characterized by (LSK) cells. Propranolol was administered for 7, 14, or 21 days. The duration of propranolol pretreatment does not lead to the improvement of GPC-100 recruitment. 16g ) CD34-LSK cell mobilization profile. Fig. 16h ) LSK cell mobilization profile. Pro: Propranolol. Data are expressed as mean ± SEM. Fig. 17 ... demonstrates in vivo LT-LSK cell mobilization. Mice were treated with propranolol (20 mg / kg, IP) or PBS (IP) for 7 days, G-CSF (0.1 mg / kg, SC) was administered twice daily from days 2 to 6, and GPC-100 (30 mg / kg, IV) was co-administered with propranolol on day 7. Blood was collected after 2 hours. Sca-1 + and cKit + Cells were gated from lineage-negative cells (LSK cells). Subsequently, LSK CD34 - CD150 using as the upper gate + and CD48 + Cells were selected. The organ HSC population is CD34 - CD150 + CD48 + It was defined as an LSK cell. Figs. 18a-18c It demonstrates the functional capacity of cells mobilized by the triple combination. (Fig. 18a) Total number of CFUs in the mobilized PB of the mouse, (Fig. 18b) CD150 +LSK cell number and (Fig. 18c) CD34 - Number of LSK cells. The total number of mobilized cells was normalized based on the WBC count after treatment. Flow cytometry and CFU analysis were performed on different cohorts of age- and body-weight-matched mice. Each mouse was analyzed individually. Pro: Propranolol, G: G-CSF. Data are expressed as mean ± SEM. Figs. 19a-19c This shows the time course of WBC subgroup mobilization by GPC-100 (Burixafor, or TG-0054), AMD3100 (Plerixafor, or Mozobil), and BL8040 (Moticaportide) in mice. Figs. 20a-20c This shows that the superiority of GPC-100 against BL8040 is specifically enhanced in the balb / c mouse strain. Fig. 21 It shows that the mobilization of myeloid cells (non-lymphocytes, C11b+ or F4 / 80+ cells) by GPC-100 is further improved by the addition of 7 days of propranolol pretreatment. Figs. 22a-22b is naive(na Compared to ve) mice, tumor-bearing mice (CT26-colon cancer) show greater mobilization, particularly for Cd11b+ myeloid cells. Figs. 23a-23b Figure 23 shows the mobilization kinetics of CD34+ cells in PB of MM patients after GPC-100 administration. Figure 23a shows the average mobilization at each time point for all patients. Figure 23b shows the relative change in CD34+ cells for each patient at each time point, and the percentage of peak relative change for all 9 patients was within the range of 45 minutes ± 15 minutes (or about 30 minutes to about 60 minutes). Fig. 24 ±2×10 by all MM patients who received combination therapy with GPC-100, propranolol, and G-CSF 6 It shows that the primary endpoint of CD34+ cells / kg has been met. Specific details for implementing the invention

[0005] CXCR4 is a member of the chemokine G protein-coupled receptor (GPCR) family [20, 21] and is expressed on HSCs [20, 22]. CXCR4 signaling, mediated by the natural ligand CXCL12, plays a pivotal role in cellular chemotaxis as well as in the maintenance and survival of HSCs in the bone marrow

[22] . GPC-100, also known as burixafor or TG-0054, is a novel small molecule antagonist with high binding affinity for CXCR4. In a Phase 1 study in healthy volunteers, GPC-100 was well tolerated, and circulating CD34 compared to baseline after a single intravenous injection + It induced a 3 to 12-fold increase in HSCs

[23] . GPC-100 was clinically tested as an HSC mobilizer in MM patients in combination with G-CSF (NCT02104427)

[24] , and >5.0×10⁻¹⁰ in 1-2 leukapheresis sessions 6 CD34 + It was shown to lead to a significant increase in HSCs per kg cell

[25] . This result was comparable to previous results obtained from G-CSF and AMD3100 treatments.

[0006] Previous studies have demonstrated that CXCR4 physically interacts with the beta-2-adrenergic receptor or β2AR (gene ADRB2) in cell systems overexpressing both receptors [26-28]. The results of Nakai et al. suggest that β2AR selectively forms a heteromeric complex with CXCR4, and that stimulation by β2AR enhances CXCR4 signaling, potentially increasing lymphocyte retention in lymph nodes and decreasing recruitment to the PB

[26] . β2AR is also expressed on HSCs [29, 30], and adrenergic signaling plays a key role in regulating the HSC niche in the BM [31, 32]. Epinephrine and norepinephrine, natural ligands of β2AR, have been shown to affect the turnover and trafficking of HSCs

[33] , as well as reduce the proliferative and differentiation capabilities of HSCs

[34] . When human HSCs were co-stimulated with G-CSF and β2AR agonists, CXCR4 expression on the HSCs increased, suggesting that the interaction between β2AR agonists and G-CSF in the BM niche promotes HSC maintenance by CXCR4 and inhibits recruitment by G-CSF

[30] .

[0007] Studies have noted the association between the use of beta-adrenergic inhibitors (beta-blockers) and positive survival outcomes in various cancer types, including MM [35, 36]. The MM microenvironment is known to induce dysregulation of HSC function, leading to changes in gene expression and altered hematopoietic differentiation [37, 38]. The effect of beta-adrenergic blockade on HSC differentiation in MM was evaluated using propranolol, an FDA-approved non-selective beta-blocker with a safe side effect profile

[39] . A biomarker-based phase 2 randomized study found that in MM patients undergoing ASCT, propranolol [induced] cell differentiation to deviate from myeloid-lineage bias and CD34 + It was shown that it shifted the profile toward an HSC-like profile, which led to improved engraftment

[38] . In this study, activation of the sympathetic nervous system shifted the basal gene expression profile toward a more inflammatory pattern named Conserved Transcriptional Response to Adversity (CTRA), which is associated with poor outcomes in ASCT

[38] . Propranolol was shown to reduce the CTRA gene signature. In another study, BM samples from MM patients showed that propranolol could enhance the differentiation of HSCs into megakaryocyte-erythrocyte progenitor cells and reduce the number of granulocyte-monocyte progenitor cells, which are known to contribute to the pro-tumorigenic niche

[40] . In summary, these studies indicate that propranolol can block the negative effects of adrenergic signaling on HSC biology and induce HSC proliferation and differentiation, and can also synergize with CXCR4 inhibitors considering the possible crosstalk between β2AR and CXCR4 in BM. Therefore, co-inhibition of the CXCR4 and β2AR pathways can improve HSC recruitment.

[0008] This technology presents the in vitro characterization and in vivo mobilization efficacy of GPC-100 compared to AMD3100, and provides the first evidence that the two receptors co-localize and exhibit functional synergy in cancer cells endogenously expressing CXCR4 and β2AR. Furthermore, this technology demonstrates enhanced mobilization by GPC-100 combined with propranolol and presents a novel strategy for clinical application in stem cell mobilization.

[0009] Terms used in this specification " progenitor cell " refers to cells capable of forming differentiated hematopoietic or myeloid cells in response to specific stimuli. The presence of progenitor cells may be evaluated by the ability of cells in a sample to form various types of colony-forming units, including, for example, CFU-GM (colony-forming unit, granulocyte-macrophage); CFU-GEMM (colony-forming unit, pluripotent); BFU-E (burst-forming unit, erythrocyte lineage); HPP-CFC (highly proliferative colony-forming cell); or other types of differentiated colonies obtainable in culture using known protocols.

[0010] "as used in this specification" stem The cells are a less differentiated form than progenitor cells. Typically, these cells are often positive for CD34. However, some stem cells do not contain this marker. These CD34 + Cells can be analyzed using Fluorescence Activated Cell Sorting (FACS), and thus their presence in a sample can be evaluated using this technique. Generally, CD34 + Cells exist only at low levels in the blood but in large quantities in the bone marrow. While other types of cells, such as endothelial cells and mast cells, can also display this marker, CD34 is considered an indicator of the presence of stem cells.

[0011] Terms used in this specification " CXCR4" refers to CXC motif chemokine receptor 4, also identified by a unique database identifier (ID) and alternative names as shown in Table 1 (Chatterjee et al., 2014; Debnath et al., 2013; Domanska et al., 2013; Guo et al., 2016; Peled et al., 2012; Roccaro et al., 2014; Walenkamp et al., 2017).

[0012]

[0013]

[0014]

[0015] * GCID: Genecards Identification

[0016] HGNC: HUGO Gene Nomenclature Committee

[0017] Terms used in this specification " GPCRx" refers to the GPCRs used in this study to investigate whether these GPCRs interact with CXCR4 and whether they exhibit characteristics distinct from those of individual protomers. Examples of GPCRx, their alternative names, and unique database identifiers (IDs) are shown in Table 1, which include ADCYAP receptor type I (ADCYAP1R1), adenosine A2b receptor (ADORA2B), adenosine A3 receptor (ADORA3), adrenergic receptor beta 2 (ADRB2), apelin receptor (APLNR), complement C5a receptor 1 (C5AR1), calcitonin receptor (CALCR), chemokine (CC motif) receptor 5 (CCR5), receptor 5 CC motif), cholinergic receptor muscarinic 1 (CHRM1), galanin receptor 1 (GALR1), endothelin receptor type B (EDNRB), histamine receptor H1 (HRH1), motilin receptor (MLNR), and neurotensin It includes receptor 1 (NTSR1), prostaglandin E receptor 2 (PTGER2), prostaglandin E receptor 3 (PTGER3), somatostatin receptor 2 (SSTR2), and tachykinin receptor 3 (TACR3).

[0018] Table 1 also includes the nomenclature of CXCR4, as well as the synergistic Ca2 upon co-stimulation with the two agonists. + To enhance the reaction, a nomenclature for GPCRx that forms a heteromer with CXCR4 is provided.

[0019] Terms used in this specification " GPC-100 "(Also known as Burixapor, TG-0054) has the chemical formula C 27 H 51It refers to the chemical compound 2-[4-[6-amino-2-[[4-[[3-(cyclohexylamino)propylamino]methyl]cyclohexyl]methylamino]pyrimidine-4-yl]piperazine-1-yl]ethylphosphonic acid having N8O3P. A representation of the chemical structure is shown in FIG. 1a. An exemplary form of GPC-100 used herein is GPC-100 hydrobromide.

[0020] Terms used in this specification " AMD3100 "(Also known as plericsaphor, or mozobil) has the chemical formula C 28 H 54 It refers to the chemical compound 1-[[4-(1,4,8,11-tetraazacyclotetradec-1-ylmethyl)phenyl]methyl]-1,4,8,11-tetraazacyclotetradecane having N8. An exemplary form of AMD3100 as used herein is plericsaphor octahydrochloride.

[0021] Terms used in this specification " beta-adrenergic receptor inhibitor " refers to a compound or pharmaceutical composition that effectively blocks the activation of beta-adrenergic receptors. As used herein, beta-adrenergic receptors refer to one of three types of beta-adrenergic receptors: adrenergic beta-1 receptors, adrenergic beta-2 receptors, and adrenergic beta-3 receptors. An exemplary beta-adrenergic receptor inhibitor used herein is propranolol.

[0022] Terms used in this specification " ADRB2 inhibitors " refers to a compound or pharmaceutical composition that effectively blocks the activation of the adrenergic beta-2 receptor (ADRB2). An exemplary ADRB2 inhibitor used herein is propranolol.

[0023] Terms used in this specification " Propranolol"(Also known as Anaprilin, Anapriline, Avlocardyl, AY 20694, AY-20694, AY20694, Betadren, Dexpropranolol, Dociton, Hydrochloride, Propranolol, Inderal, Obsidan, Obzidan, Propanolol, Propranolol, Propranolol Hydrochloride, Rexigen, etc.) is chemical formula C 16 H 21 It refers to the chemical compound 1-naphthalene-1-yloxy-3-(propan-2-ylamino)propan-2-ol having NO2. An exemplary form of propranolol used herein is propranolol hydrochloride.

[0024] Terms used in this specification " G-CSF "(GCSF, also known as CSF 3) refers to granulocyte colony-stimulating factor, a glycoprotein known to function as a cytokine and hormone. As used herein, the term G-CSF includes all variants, forms, structural analogs, functional analogs, derivatives, biosimilars, etc. of the natural glycoprotein. The term G-CSF also includes any recombinant form of G-CSF. Analogs of G-CSF may include filgrastim and lenograstim.

[0025] Terms used in this specification " Lenalidomide "(Also known as Revlimid) has the chemical formula C 13 H 13 It refers to the chemical compound 3-(7-amino-3-oxo-1H-isoindole-2-yl)piperidin-2,6-dione containing N3O3. Lenalidomide can be administered to treat multiple myeloma.

[0026] Terms used in this specification " Daratumumab"(Also known as Darzalex) refers to a monoclonal antibody that binds to CD38. As used herein, the term daratumumab includes all variants, forms, structural analogs, functional analogs, derivatives, biosimilars, etc. of monoclonal antibodies approved by relevant regulatory agencies for the treatment of cancer."

[0027] Terms used in this specification " apheresis "It is a medical procedure that passes a subject's blood through a device that separates one specific component from the blood and returns the remainder to the subject. Apheresis and specific types of apheresis. In leukocyte apheresis, leukocytes are separated from a blood sample. Exemplary apheresis techniques include (1) plasma exchange, (2) LDL apheresis, (3) photopheresis, (4) immunosorbent using a Staphylococcal protein A-agarose column, (5) leukocytapheresis, (6) erythrocytapheresis, (7) thrombocytapheresis, and (8) leukocyte apheresis. Apheresis is a difficult, inconvenient, and expensive procedure.

[0028] Terms used in this specification " Leukocyte apheresis It is a medical procedure and a specific type of apheresis. In leukocyte apheresis, white blood cells are separated from a blood sample.

[0029] Terms used in this specification " inhibitor" refers to a molecule that inhibits or suppresses the enhanced function of a biological molecule. For example, the biological molecule may be CXCR4, a beta-adrenergic receptor, a GPCR, a heteromer of CXCR4 and a beta-adrenergic receptor, a CXCR4-GPCRx heteromer, a CXCR4 homodimer, and / or a beta-adrenergic receptor homodimer. Non-limiting examples of inhibitors of the present invention that may be used for cell recruitment include GPCRx antagonists, GPCRx inverse agonists, GPCRx positive and negative allosteric modulators, antigen-binding portions thereof comprising CXCR4-GPCRx heteromer-specific antibodies or single-domain antibody-like scaffolds, and divalent ligands having a pharmacophore selective for CXCR4 connected by a spacer arm to a pharmacophore selective for GPCRx. It includes bispecific antibodies against CXCR4 and GPCRx, a radiolabeled CXCR4 ligand linked to a GPCRx ligand, and a small molecule ligand that inhibits heteromer-selective signaling. Upon co-stimulation with the two agents, it forms a heteromer with CXCR4 and Ca2 + Specific examples of inhibitors for GPCRx that enhance the response are listed in Table 2. Non-limiting examples of inhibitors may be natural or artificial and may include small molecule inhibitors; peptide inhibitors; antibodies, fragments thereof, or similar substances; and aptamers. Inhibitors may be antagonists.

[0030] Terms used in this specification " antagonist"It refers to a type of receptor ligand or drug that blocks or weakens a biological response by binding to and blocking a receptor, and is also called a blocker. Antagonists have an affinity for cognate receptors but lack efficacy; their binding interferes with interactions and inhibits the function of agonists or counter-agonists at cognate receptors. Upon co-stimulation with two agonists, it forms a heteromer with CXCR4 and Ca2 + Specific examples of antagonists for GPCRx that enhance the response are listed in Table 2.

[0031]

[0032] Terms used in this specification " heteromer"Heteromer" refers to a macromolecular complex composed of different macromolecules or macromolecular variants that combine to have one or more functions as a single unit. Different macromolecules or macromolecular variants may have distinct structure(s) and function(s). In contrast, a "homomer" is a macromolecular complex composed of one or more identical macromolecules. Like heteromers, the macromolecules of a homomer combine to have one or more functions as a single unit. When referring to GPCRs, the two GPCR units [protomers] have biochemical properties that are distinctly different from the biochemical properties of their individual components. Heteromerization and homomerization can be evaluated by co-localization analysis based on immunochemistry, microscopy, etc.; DNA / RNA analysis; resonance energy transfer analysis based on fluorescence, bioluminescence, etc.; chromatography; crystallography; mass spectrometry; nuclear magnetic resonance spectroscopy, etc. An exemplary method for evaluating heteromerization and homomerization is in situ hybridization (in It may include situ hybridization), immunohistochemistry, RNAseq, reverse transcription-quantitative PCR (RT-qPCR, real-time PCR), microarray, proximity ligation analysis (PLA), time-resolved FRET (TR-FRET), whole-body single-photon emission computed tomography (SPECT) or positron emission tomography / computed tomography (PET / CT), cryo-electron microscopy (cryo-EM), brightness / number analysis, fluorescence correlation spectroscopy (FCS), fluorescence cross-correlation spectroscopy (FCCS), and / or similar techniques.

[0033] Phrases used in this specification " Effective amount "An effective dose refers to an amount sufficient to produce a beneficial or desired result. The effective dose may be administered as one or more doses, applications, or dosages. Such delivery depends on a number of variables, including the duration of use of individual dose units, the bioavailability of the formulation, and the route of administration.

[0034] Phrases used in this specification " Therapeutic effective dose " refers to an amount of therapeutic agent (e.g., inhibitors, antagonists, or any other therapeutic agent provided herein) sufficient to reduce, improve, and / or prevent the severity and / or duration of cancer and / or related symptoms. The therapeutically effective amount of therapeutic agent may be an amount necessary to reduce, improve, or prevent the progression or advancement of cancer, reduce, improve, or prevent the recurrence, occurrence, or onset of cancer, and / or improve or enhance the prophylactic or therapeutic effects of other therapies (e.g., therapies other than the administration of inhibitors, antagonists, or any other therapeutic agent provided herein)."

[0035] Phrase " remedy " refers to any agent that may be used for the treatment, improvement, prevention, or management of cancer and / or related symptoms. In some embodiments, the therapeutic agent refers to an inhibitor of the CXCR4-GPCRx heteromer of the present invention. The therapeutic agent may be an agent that is well known, has been used, or is currently used as being useful for the treatment, improvement, prevention, or management of cancer and / or related symptoms.

[0036] Phrases used in this specification " Intracellular Ca2 + analyze ", " Calcium Mobilization Analysis Or variations thereof refer to cell-based assays for measuring calcium flux associated with GPCR activation or inhibition. The method utilizes a calcium-sensitive fluorescent dye that is absorbed into the cytoplasm. The dye binds to calcium released from intracellular stores, increasing its fluorescence. Changes in fluorescence intensity are directly correlated with the amount of intracellular calcium released into the cytoplasm in response to ligand activation of the receptor of interest.

[0037] Phrases used in this specification " proximity-based assay"This refers to biophysical and biochemical techniques capable of monitoring the proximity and / or binding of two protein molecules in vitro (in cell lysates) and in living cells, including bioluminescent resonance energy transfer (BRET), fluorescence resonance energy transfer (FRET), bimolecular fluorescence complementation (BiFC), proximity ligation assay (PLA), cysteine ​​crosslinking, co-immunoprecipitation (Ferre et al., 2009; Gomes et al., 2016), fluorescence correlation spectroscopy (FCS), fluorescence cross-correlation spectroscopy (FCCS), proximity-dependent biotin identification, proximity extension assay, split-luciferase assay, proximity-dependent tethered biotinylation, etc.

[0038] The following terms used in this specification have the specified meanings.

[0039] Term " About "is as understood by those skilled in the art" approximately It has a plain and general meaning. When the term "approximately" is used with a numerical range, it modifies the range by extending the boundaries above and below the presented numerical value. Generally, the terms "approximately" or "roughly" are used to modify a numerical value by a deviation of 20% above or below the specified value.

[0040] Term " to include (comprise) ", " including (comprising) ", " contain ", " containing ", " include ", " including ", " Includes, but not limited to " or " Characterized by It is comprehensive or open and does not exclude additional unlisted elements.

[0041] Terms used in this specification " administer ", " administering ", " Administered ", " administration of"etc. refers to the administration of a substance to a subject in need through any pharmacologically appropriate route. Common routes of administration of a substance to a subject include oral, intravenous, intramuscular, subcutaneous, topical, transdermal, inhalation, sublingual, oral, rectal, nasal, ocular, intrathecal, epidural, vaginal, intra-articular, intradermal, intratumoral, and combinations thereof.

[0042] Terms used in this specification " treatment " or " treating "The rest means a specific method, process, composition (e.g., drug, device, or chemical, biochemical, or biological material) or combination thereof for managing, curing, alleviating, or preventing a specific disease, condition, or symptom of a subject. The term includes therapeutic, preventive, and palliative interventions aimed at improving the health or quality of life of an individual or group. Treatment may include combination therapy, multistep therapy, and multiphase therapy.

[0043] Phrases used in this specification " qualify for treatment " or " Qualifying for treatment " or " to qualify for treatment " or " Qualifying ... for treatment "etc. refers to a specific method, process, composition (e.g., drug, device, or chemical, biochemical, or biological material) or combination thereof for qualifying a subject for treatment or satisfying preconditions for treatment. The subject may ultimately undergo treatment, or the subject may ultimately not receive treatment.

[0044] Phrases used in this specification " prepare for treatment " or " Preparing for treatment " or " to prepare … for treatment or " Preparing … for treatment"etc." means a specific method, process, composition (e.g., drug, device, or chemical, biochemical, or biological substance) or combination thereof for qualifying a subject for the treatment the subject ultimately receives.

[0045] As used in this specification and the appended claims, the following general rules apply.

[0046] singular form " one(a) ", " one(an) " and " The " includes plural references unless the content clearly indicates otherwise.

[0047] As used herein, a list of numerical ranges for a variable is intended to convey that the present invention may be practiced using the variable as any value within the said range. Thus, in the case of an essentially discrete variable, the variable may be equal to any integer value in the numerical range, including the endpoint of the range. Similarly, in the case of an essentially continuous variable, the variable may be equal to any real value in the numerical range, including the endpoint of the range. For example, a variable described as having a value between 0 and 10 may be 0, or 1, or 2, or 3…, or 10 in the case of an essentially discrete variable, and 0.0, 0.1, 0.01, 0.001, or any other real value in the case of an essentially continuous variable.

[0048] Unless otherwise indicated, this description adopts conventional chemical, biochemical, molecular biology, immunology, cancer biology, and pharmacological methods and terms that have a general meaning to those skilled in the art. All publications, references, patents, and patent applications cited herein are incorporated herein by reference in their entirety.

[0049] The present specification discloses methods and compositions for mobilizing cells in a subject by blocking CXCR4, beta-adrenergic receptors, GPCRs, or any combination thereof. In some embodiments, the cells are stem cells. In some embodiments, the cells are immune cells. In some embodiments, mobilizing cells in a subject comprises blocking CXCR4. In some embodiments, mobilizing cells in a subject comprises blocking beta-adrenergic receptors. In some embodiments, mobilizing cells in a subject comprises blocking GPCRs. In some embodiments, mobilizing cells in a subject comprises blocking CXCR4 and beta-adrenergic receptors. In some embodiments, mobilizing cells in a subject comprises blocking CXCR4 and GPCRs. In some embodiments, mobilizing cells in a subject comprises blocking CXCR4-GPCR heteromers.

[0050] The present specification discloses a method for mobilizing cells in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. The present specification also discloses a method for inducing cell mobilization in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. In some embodiments, blocking of beta-adrenergic receptor signaling is performed before blocking CXCR4 signaling. In some embodiments, blocking of beta-adrenergic receptor signaling is performed at a first specific time interval before blocking CXCR4 signaling. In some embodiments, the first specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all. In some embodiments, the blocking of beta-adrenergic receptor signaling continues even after the blocking of CXCR4 signaling has ended. In some embodiments, the blocking of beta-adrenergic receptor signaling continues for a second specific time interval after the blocking of CXCR4 signaling has ended.In some embodiments, the second specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all.

[0051] In some embodiments, blocking of CXCR4 signaling involves administering a CXCR4 inhibitor to a subject.

[0052] In some embodiments, blocking of beta-adrenergic receptor signaling involves administering a beta-adrenergic receptor inhibitor to a subject. In some embodiments, blocking of CXCR4 signaling involves administering a CXCR4 inhibitor to a subject, and blocking of beta-adrenergic receptor signaling involves administering a beta-adrenergic receptor inhibitor to a subject. In some embodiments, the cell is a stem cell. In some embodiments, the cell is an immune cell.

[0053] The present specification discloses a method for mobilizing stem cells in a subject, comprising the step of administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject. The present specification also discloses a method for inducing stem cell mobilization in a subject, comprising the step of administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject. In some embodiments, the administration of the beta-adrenergic receptor inhibitor is performed before the administration of the CXCR4 inhibitor. In some embodiments, the administration of the beta-adrenergic receptor inhibitor is performed at a first specific time interval before the administration of the CXCR4 inhibitor. In some embodiments, the first specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all. In some embodiments, administration of the beta-adrenergic receptor inhibitor continues even after administration of the CXCR4 inhibitor has ended. In some embodiments, administration of the beta-adrenergic receptor inhibitor continues for a second specific time interval after administration of the CXCR4 inhibitor has ended.In some embodiments, the second specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all.

[0054] In some embodiments, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor. In some embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenolol, atenolol, betaxolol, bupranolol, butoxamine, carazolol, carvedilol, CGP 12177, cicloprolol, ICI 118551, ICYP, labetalol, levobetaxolol, levobunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, and timolol. In some embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551. In some embodiments, the beta-adrenergic receptor inhibitor is propranolol.

[0055] In some embodiments, the CXCR4 inhibitor is ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549, D-[Lys3]GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, Isothiourea-1a, Isothiourea-1t (IT1t), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[11C]methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, GPC-100 (burixafor, or TG-0054), USL311, viral macrophage inflammation protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]O2-AMD3100, [177Lu]pentixather, ulocuplumab (MDX1338 / BMS-936564), AD-214, LY2624587, PF-06747143, POL6326, It is selected from the group consisting of MB1707 and 508MC1 (compound 26). GPC-100 is also referred to as burixaphor or TG-0054. AMD3100 is also referred to as plerixaphor or mozobil.In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, GPC-100 (Bulixafor, or TG-0054), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is GPC-100 (Bulixafor, or TG-0054). In some embodiments, the CXCR4 inhibitor is AMD3100 (plerixafor, or mozobil). In some embodiments, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564).

[0056] In some embodiments, administration of a CXCR4 inhibitor to a subject includes administration of GPC-100 (burixafor, or TG-0054) and propranolol. In some embodiments, administration of a CXCR4 inhibitor to a subject includes administration of AMD3100 (plerixafor, or mozovil) and propranolol. In some embodiments, administration of a CXCR4 inhibitor to a subject includes administration of ulocuplumab (MDX1338 / BMS-936564) and propranolol.

[0057] In some embodiments, the method further comprises the step of administering G-CSF to a subject. In some embodiments, the administration of a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to a subject is performed in the absence of G-CSF. The present specification discloses a method for mobilizing stem cells in a subject, comprising the step of administering a CXCR4 inhibitor and G-CSF to a subject in the absence of a beta-adrenergic receptor inhibitor. The present specification also discloses a method for inducing stem cell mobilization in a subject, comprising the step of administering a CXCR4 inhibitor and G-CSF to a subject in the absence of a beta-adrenergic receptor inhibitor. In some embodiments, the administration of a CXCR4 inhibitor to a subject comprises the administration of GPC-100 (burixafor, or TG-0054) and G-CSF. In some embodiments, the administration of a CXCR4 inhibitor to a subject comprises the administration of AMD3100 (plerixafor, or mozobil) and G-CSF. In some embodiments, administration of a CXCR4 inhibitor to a subject includes administration of ulocuplumab (MDX1338 / BMS-936564) and G-CSF.

[0058] The present specification also discloses a method for inducing stem cell mobilization in a subject, comprising the step of administering a CXCR4 inhibitor, G-CSF, and a beta-adrenergic receptor inhibitor. In some embodiments, administration of the CXCR4 inhibitor to the subject comprises administration of GPC-100 (burixafor, or TG-0054), G-CSF, and propranolol. In some embodiments, administration of the CXCR4 inhibitor to the subject comprises administration of AMD3100 (plerixafor, or mozovil), G-CSF, and propranolol. In some embodiments, administration of the CXCR4 inhibitor to the subject comprises administration of ulocuplumab (MDX1338 / BMS-936564), G-CSF, and propranolol.

[0059] In some embodiments, administration of a combination of a CXCR4 inhibitor and G-CSF induces an enhanced amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone. In some embodiments, administration of a combination of a CXCR4 inhibitor and G-CSF mobilizes cells in an enhanced amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 1.1 to 1.2 times, 1.2 to 1.3 times, 1.3 to 1.4 times, 1.4 to 1.5 times, 1.5 to 1.6 times, 1.6 to 1.7 times, 1.7 to 1.8 times, 1.8 to 1.9 times, 1.9 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 4 times, 4 to 5 times, 5 to 10 times, or 10 times or more. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 5%-10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%-120% more, 120%-140% more, 140%-160% more, 160%-180% more, 180%-200% more, 200%-250% more, or 250%-300% more, 300%-400% more, 400%-500% more, 500%-750% more, 750%-1000% more, or 1000% or more.

[0060] In some embodiments, administration of a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone. In some embodiments, administration of a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 1.1 to 1.2 times, 1.2 to 1.3 times, 1.3 to 1.4 times, 1.4 to 1.5 times, 1.5 to 1.6 times, 1.6 to 1.7 times, 1.7 to 1.8 times, 1.8 to 1.9 times, 1.9 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 4 times, 4 to 5 times, 5 to 10 times, or 10 times or more. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 5%-10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%-120% more, 120%-140% more, 140%-160% more, 160%-180% more, 180%-200% more, 200%-250% more, or 250%-300% more, 300%-400% more, 400%-500% more, 500%-750% more, 750%-1000% more, or 1000% or more.

[0061] In some embodiments, administration of a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor alone. In some embodiments, administration of a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor alone. In some embodiments, administration of a combination of AMD3100 (Plerixafor, or Mozobil) and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by AMD3100 (Plerixafor, or Mozobil) and G-CSF. In some embodiments, administration of a combination of AMD3100 (Plerixafor, or Mozobil) and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by AMD3100 (Plerixafor, or Mozobil) and G-CSF. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 1.1 to 1.2 times, 1.2 to 1.3 times, 1.3 to 1.4 times, 1.4 to 1.5 times, 1.5 to 1.6 times, 1.6 to 1.7 times, 1.7 to 1.8 times, 1.8 to 1.9 times, 1.9 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 4 times, 4 to 5 times, 5 to 10 times, or 10 times or more.In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 5%-10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%-120% more, 120%-140% more, 140%-160% more, 160%-180% more, 180%-200% more, 200%-250% more, or 250%-300% more, 300%–400% more, 400%–500% more, 500%–750% more, 750%–1000% more, or 1000% or more. In some embodiments, the increased amount of cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) analysis. In some embodiments, the increased amount of cell mobilization or apheresis is measured by flow cytometry. In some embodiments, flow cytometry is (Lin. - Sca1 + c-Kit + ) is performed on LSK cells. In some embodiments, flow cytometry is performed on CD34 - It is performed on LSK cells. In some embodiments, flow cytometry is performed on CD150 + It is performed on LSK cells. In some embodiments, flow cytometry is performed on CD48 + It is performed on cells. In some embodiments, the increased amount of cell mobilization or component collection is measured by colony-forming unit (CFU) analysis.

[0062] In some embodiments, administration of a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor and G-CSF alone. In some embodiments, administration of a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor and G-CSF alone. In some embodiments, administration of a combination of GPC-100 (burixafor, or TG-0054), G-CSF, and a beta-adrenergic receptor inhibitor induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF. In some embodiments, administration of a combination of GPC-100 (burixafor, or TG-0054), G-CSF, and a beta-adrenergic receptor inhibitor mobilizes cells in an increased amount compared to the amount of cell mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 1.1 to 1.2 times, 1.2 to 1.3 times, 1.3 to 1.4 times, 1.4 to 1.5 times, 1.5 to 1.6 times, 1.6 to 1.7 times, 1.7 to 1.8 times, 1.8 to 1.9 times, 1.9 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 4 times, 4 to 5 times, 5 to 10 times, or 10 times or more.In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 5%-10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%-120% more, 120%-140% more, 140%-160% more, 160%-180% more, 180%-200% more, 200%-250% more, or 250%-300% more, 300%–400% more, 400%–500% more, 500%–750% more, 750%–1000% more, or 1000% or more. In some embodiments, the increased amount of cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) analysis. In some embodiments, the increased amount of cell mobilization or apheresis is measured by flow cytometry. In some embodiments, flow cytometry is (Lin. - Sca1 + c-Kit + ) is performed on LSK cells. In some embodiments, flow cytometry is performed on CD34 - It is performed on LSK cells. In some embodiments, flow cytometry is performed on CD150 + It is performed on LSK cells. In some embodiments, flow cytometry is performed on CD48 + It is performed on LSK cells. In some embodiments, the increased amount of cell mobilization or component aggregation is measured by colony-forming unit (CFU) analysis.

[0063] In some embodiments, administration of a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induces an increased amount of long-term repopulating hematopoietic stem cell (LT-HSC) recruitment compared to the amount of LT-HSC recruitment induced by the CXCR4 inhibitor and G-CSF alone. As used herein, LT-HSCs are CD150 + , CD34 - , or CD48 + Includes LSK cells. In some embodiments, administration of a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and a G-CSF mobilizes LT-HSCs in an increased amount compared to the amount of LT-HSC mobilization induced by the CXCR4 inhibitor and G-CSF alone. In some embodiments, administration of a combination of GPC-100 (burixafor, or TG-0054), G-CSF, and a beta-adrenergic receptor inhibitor induces an increased amount of LT-HSC mobilization compared to the amount of LT-HSC mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF. In some embodiments, administration of a combination of GPC-100 (burixafor, or TG-0054), G-CSF, and a beta-adrenergic receptor inhibitor mobilizes LT-HSCs in an increased amount compared to the amount of LT-HSC mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF. In some embodiments, the increased amount of LT-HSC mobilization compared to the amount of LT-HSC mobilization induced by the CXCR4 inhibitor alone is 1.1 to 1.2 times, 1.2 to 1.3 times, 1.3 to 1.4 times, 1.4 to 1.5 times, 1.5 to 1.6 times, 1.6 to 1.7 times, 1.7 to 1.8 times, 1.8 to 1.9 times, 1.9 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 4 times, 4 to 5 times, 5 to 10 times, or 10 times or more.

[0064] In some embodiments, the subject has a CXCR4 protomer within the cell. In some embodiments, the subject has an ADRB2 protomer within the cell. In some embodiments, the subject has a CXCR4 protomer and an ADRB2 protomer within the cell. In some embodiments, the subject has a CXCR4-ADRB2 heteromer within the cell. In some embodiments, i) the CXCR4-ADRB2 heteromer has an increased amount of downstream calcium mobilization compared to downstream calcium mobilization from the CXCR4 protomer or the ADRB2 protomer; and ii) the administered inhibitor combination inhibits the enhanced downstream calcium mobilization from the CXCR4-ADRB2 heteromer within the stem cell.

[0065] In some embodiments, the cell is a stem cell. In some embodiments, the stem cell is selected from the group consisting of hematopoietic stem cells (HSC), hematopoietic progenitor cells (HPC), mesenchymal stem cells, endothelial progenitor cells, neural stem cells, epithelial stem cells, skin stem cells, and cancer stem cells. In some embodiments, the stem cell is an HSC or HPC. In some embodiments, the HSC or HPC is mobilized from the bone marrow into peripheral blood (PB). In some embodiments, the mobilized HSC or HPC is collected for transplantation into a cancer patient. In some embodiments, the cancer is selected from the group consisting of lymphoma, leukemia, and multiple myeloma. In some embodiments, the cancer is non-Hodgkin's lymphoma (NHL), Hodgkin's Disease (HD) or Hodgkin's Lymphoma (HL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), multiple myeloma (MM) or Burkitt's Lymphoma. In some embodiments, the stem cells are mesenchymal stem cells. In some embodiments, the mesenchymal stem cells are mobilized from the bone marrow into the peripheral blood.In some embodiments, mesenchymal stem cells are mobilized for the treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the stem cells are cancer stem cells. In some embodiments, the cancer stem cells are mobilized into the blood. In some embodiments, the cancer stem cells are mobilized for the treatment of cancer.

[0066] In some embodiments, the cell is an immune cell. In some embodiments, the immune cell is a white blood cell (WBC). In some embodiments, the white blood cell is a lymphocyte. In some embodiments, the lymphocyte is selected from the group consisting of T cells, B cells, and natural killer (NK) cells. In some embodiments, the lymphocyte is a T cell. In some embodiments, the lymphocyte is a natural killer (NK) cell. In some embodiments, the white blood cell is a granulocyte. In some embodiments, the granulocyte is selected from the group consisting of neutrophils, eosinophils, and basophils. In some embodiments, the granulocyte is a neutrophil. In some embodiments, the white blood cell is a monocyte. In some embodiments, the immune cell is mobilized from the bone marrow to the peripheral blood. In some embodiments, the immune cell is mobilized from the lymph nodes to the peripheral blood. In some embodiments, the mobilized immune cell is used in adoptive cell therapy (ACT). In some embodiments, adoptive cell therapy (ACT) is chimeric antigen receptor (CAR) T cell therapy. In some embodiments, adoptive cell therapy (ACT) is natural killer (NK) cell therapy. In some embodiments, adoptive cell therapy (ACT) is engineered T-cell receptor (TCR) therapy. In some embodiments, adoptive cell therapy (ACT) is tumor-infiltrating lymphocyte (TIL) therapy.

[0067] In some embodiments of the present invention, mobilizing cells in a subject involves blocking CXCR4. Many antiviral agents that inhibit HIV replication by inhibiting CXCR4, a co-receptor required for the fusion and entry of T-tropic HIV strains, also inhibit binding and signaling induced by the natural ligand chemokine CXCL12 (also known as SDF-1). Without being bound by theory, agents that inhibit the binding of CXCL12 to CXCR4 may result in increased mobilization of stem and / or progenitor cells to the periphery due to this inhibition. Enhancing the mobilization of stem and / or progenitor cells to peripheral blood is beneficial for therapies to mitigate the effects of protocols that adversely affect the bone marrow, such as causing leukopenia, a known side effect of chemotherapy and radiotherapy. Agents that inhibit the binding of CXCL12 to CXCR4 also improve the success of bone marrow transplants, enhance wound healing and burn treatment, and aid in the restoration of damaged organ tissue. They fight against bacterial infections prevalent in leukemia. They use CD34 via apheresis, regardless of whether it is combined with other mobilization factors. + It is used to mobilize and harvest cells. The harvested cells are used for treatments requiring stem cell transplantation.

[0068] In some embodiments of the present invention, mobilizing stem cells from a subject involves blocking CXCR4-GPCR heteromers. Although various CXCR4-GPCR heteromers with distinct physiological and pharmacological properties have been reported, their role in stem cell mobilization or the potential for developing stem cell mobilization therapies targeting CXCR4-GPCR heteromers has not been clearly understood or recognized.

[0069] In some embodiments, the CXCR4-GPCR heteromer is blocked on cells expressing CXCR4 and β2AR receptors in close proximity. As used herein, "close proximity" includes proximity sufficient to be detectable using PLA analysis. In some embodiments, the cancer is blood cancer. In some embodiments, the cancer cells express endogenous CXCR4 and β2AR receptors in close proximity. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is Burkitt lymphoma.

[0070] In the industry, GPCRs were considered to function as monomers that interact with heterotrimeric G proteins upon ligand binding, and drugs were developed based on monomeric or homomeric GPCRs

[41] . Recently, this view has changed dramatically based on the discovery that GPCRs can form heteromers and that heteromerization is essential for some GPCRs. GPCR heteromerization is known to alter GPCR maturation and cell surface delivery, ligand binding affinity, signaling strength and pathways, as well as receptor desensitization and recycling [42-46]. Different GPCR heteromers exhibit distinct functional and pharmacological properties, and GPCR heteromerization can vary depending on cell type, tissue, disease, or pathological state [42-46]. GPCR heteromerization is now considered a common phenomenon, and deciphering GPCR heteromerization opens new avenues for understanding receptor function, physiology, and roles in disease and pathological states. Accordingly, identifying GPCR heteromers and their functional characteristics provides new opportunities to develop new drugs with fewer side effects, higher efficacy, and increased tissue selectivity, or to find new uses for existing drugs [42, 43, 45].

[0071] Apheresis is a standard practice for obtaining a larger number of immune cells as a starting material for adoptive cell therapy (ACT), a treatment based on delivering cells into a patient. Apheresis may involve passing a patient's blood through a device that separates a specific component and returns the remainder to the patient's blood circulation. Therefore, apheresis is an ex vivo therapy. Different processes are employed in apheresis depending on the substance being removed. Centrifugation is the most common method when separation by density is required. Other methods include adsorption onto beads coated with an adsorbent material and filtration. Centrifugation methods can be divided into two basic categories: continuous flow centrifugation (CFC) and intermittent flow centrifugation.

[0072] Various apheresis techniques may be used whenever the removed component causes severe symptoms of the disease in a patient. Generally, apheresis must be performed quite frequently and is an invasive procedure. Therefore, it is typically adopted when other means of controlling a specific disease have failed, or when the nature of the symptoms is such that waiting for medication to take effect entails suffering or a risk of complications. Apheresis techniques include the following: (1) Plasma exchange – the removal of the liquid portion of blood to eliminate harmful substances, in which plasma is replaced with an exchange solution; (2) LDL apheresis – the removal of low-density lipoprotein in patients with familial hypercholesterolemia; and (3) Photoapheresis – used to treat graft-versus-host disease, cutaneous T-cell lymphoma, and heart transplant rejection. (4) Immunoassay using a Staphylococcus protein A-agarose column - Removal of alloantibodies and autoantibodies (in autoimmune diseases, transplant rejection, and hemophilia) by inducing plasma through a protein A-agarose column (protein A is a cell wall component produced by various strains of Staphylococcus aureus that binds to the Fc region of IgG); (5) Leukocyte apheresis - Removal of malignant leukocytes in individuals with symptomatic very high leukocyte counts and leukemia; (6) Erythrocyte apheresis - Removal of erythrocytes in individuals with iron overload resulting from hereditary hemochromatosis or transfusion iron overload; (7) Platelet apheresis - Removal of platelets in individuals with symptoms caused by an extreme increase in platelet count, such as those with essential thrombocytosis or polycythemia vera; and (8) leukocyte apheresis - separating excess leukocytes from leukemia patients while recirculating the rest of their blood.

[0073] Apheresis is a difficult, inconvenient, and costly procedure. With the rapid growth of ACTs, including CAR-T, CAR-NK, tumor-infiltrating lymphocytes (TILs), and engineered T-cell receptors (TCRs), there is an increasing demand for apheresis techniques for the routine production of pure immune cells. The industry supplying GMP-grade starter materials for ACTs is also growing rapidly. Consequently, stem cell mobilization techniques capable of controlling immune cell types and improving apheresis yields have become important.

[0074] The enhanced stem cell mobilization (SCM) or cell mobilization methods disclosed herein can further enhance or facilitate conventional apheresis procedures. In certain embodiments, enhanced stem cell mobilization (SCM) or cell mobilization is particularly beneficial to the apheresis technique of leukocyte apheresis. In some embodiments, administration of a CXCR4 antagonist to a subject further enhances apheresis by enhancing SCM or cell mobilization. In some embodiments, administration of a beta-adrenergic receptor antagonist to a subject in combination with a CXCR4 antagonist further enhances apheresis by enhancing SCM or cell mobilization, and / or replaces the G-CSF component of the therapeutic regimen with a non-selective beta-blocker such as propranolol. In some embodiments, enhancement of SCM consequently provides benefits for HSC transplantation or CAR-T cell production for cancer immunotherapy. Currently, CXCR4 inhibitors such as AMD3100 (Plerixafor, or Mozovil), which are approved as stem cell mobilizers, are used as standard treatment in combination with G-CSF to provide concentrated HSCs and HPCs from healthy donors and are marketed as "mobilized leukopaks."

[0075] The present specification discloses a method for enhancing apheresis in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. The present specification also discloses a method for enhancing apheresis by inducing cell mobilization in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. Furthermore, the present specification discloses a method for enhancing apheresis by mobilizing cells in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. In some embodiments, blocking of beta-adrenergic receptor signaling is performed before blocking CXCR4 signaling. In some embodiments, blocking of beta-adrenergic receptor signaling is performed at a first specific time interval before blocking CXCR4 signaling. In some embodiments, the first specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all. In some embodiments, the blocking of beta-adrenergic receptor signaling continues even after the blocking of CXCR4 signaling has ended. In some embodiments, the blocking of beta-adrenergic receptor signaling continues for a second specific time interval after the blocking of CXCR4 signaling has ended.In some embodiments, the second specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all.

[0076] In some embodiments, blocking of CXCR4 signaling involves administering a CXCR4 inhibitor to a subject.

[0077] The present specification discloses a method for enhancing apheresis in a subject, comprising the step of administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to a subject. The present specification also discloses a method for enhancing apheresis by inducing cell mobilization in a subject, comprising the step of administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to a subject. Furthermore, the present specification discloses a method for enhancing apheresis by mobilizing cells in a subject, comprising the step of administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to a subject. In some embodiments, the administration of the beta-adrenergic receptor inhibitor is performed at a first specific time interval prior to the administration of the CXCR4 inhibitor. In some embodiments, the first specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all. In some embodiments, administration of the beta-adrenergic receptor inhibitor continues even after administration of the CXCR4 inhibitor has ended. In some embodiments, administration of the beta-adrenergic receptor inhibitor continues for a second specific time interval after administration of the CXCR4 inhibitor has ended.In some embodiments, the second specific time interval is 5 to 10 minutes, 10 to 20 minutes, 20 to 30 minutes, 30 to 40 minutes, 40 to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 to 2 days, 2 to 3 days, 3 to 4 days, 4 to 5 days, 5 to 6 days, 6 to 7 days, 7 to 8 days, 8 to 9 days, 9 to 10 days, 10 to 11 days, 11 to 12 days, 12 to 13 days, 13 to 14 days, or 14 days That is all.

[0078] In some embodiments, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor. In some embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenorolol, atenolol, betaxolol, bufranolol, butoxamine, carazolol, carvedilol, CGP 12177, cycloprorolol, ICI 118551, ICYP, labetalol, levobetaxolol, levovunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, and timolol. In some embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551. In some embodiments, the beta-adrenergic receptor inhibitor is propranolol.

[0079] In some embodiments, the CXCR4 inhibitor is ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549, D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, Isothiourea-1a, Isothiourea-1t (IT1t), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[11C]methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, GPC-100 (burixafor, or TG-0054), USL311, viral macrophage inflammation protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixater, [99mTc]O2-AMD3100, [177Lu]pentixater, ulocuplumab (MDX1338 / BMS-936564), AD-214, LY2624587, PF-06747143, POL6326, MB1707, and It is selected from the group consisting of 508MC1 (compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, or X4P-001), AMD3100 (Plerixapor, or Mozobil), BKT140 (BL-8040, TF14016, or 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, GPC-100 (Bulixapor, or TG-0054), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is GPC-100 (burixapor, or TG-0054).In some embodiments, the CXCR4 inhibitor is AMD3100 (plerixafor, or mozobil). In some embodiments, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564).

[0080] In some embodiments, the CXCR4 inhibitor forms an ionic interaction with Arg188 on CXCR4. In some embodiments, the CXCR4 inhibitor forms a hydrogen bond with Gln200 on CXCR4. In some embodiments, the CXCR4 inhibitor forms an ionic interaction with Arg188 on CXCR4 and forms a hydrogen bond with Gln200 on CXCR4.

[0081] In some embodiments, the CXCR4 inhibitor has a higher binding affinity for CXCL12 than AMD3100. In some embodiments, the CXCR4 inhibitor has a lower inhibitory constant (Ki) than AMD3100. In some embodiments, the CXCR4 inhibitor has an inhibitory constant that is at least 2, 5, 10, or 20 times lower than AMD3100. In some embodiments, the CXCR4 inhibitor inhibits CXCL12-induced calcium flux to a level equivalent to or higher than that of AMD3100. In some embodiments, the CXCR4 inhibitor inhibits CXCL12-induced β-arrestin recruitment to a level equivalent to or higher than that of AMD3100. In some embodiments, the CXCR4 inhibitor inhibits CXCL12-induced migration of cancer cells to a level equivalent to or higher than that of AMD3100. In some embodiments, the CXCR4 inhibitor inhibits CXCL12-induced migration of U937 and MM.1S cells to a level equivalent to or higher than that of AMD3100. In some embodiments, the CXCR4 inhibitor inhibits calcium flux induced by CXCL12 and epinephrine.

[0082] In some embodiments, administration of a CXCR4 inhibitor to a subject includes administration of GPC-100 (burixafor, or TG-0054) and propranolol. In some embodiments, administration of a CXCR4 inhibitor to a subject includes administration of AMD3100 (plerixafor, or mozovil) and propranolol. In some embodiments, administration of a CXCR4 inhibitor to a subject includes administration of ulocuplumab (MDX1338 / BMS-936564) and propranolol.

[0083] In some embodiments, the method further comprises the step of administering G-CSF to a subject. In some embodiments, the administration of a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to a subject is performed in the absence of G-CSF. The present specification discloses a method for enhancing apheresis in a subject, comprising the step of administering a CXCR4 inhibitor and G-CSF to a subject in the absence of a beta-adrenergic receptor inhibitor. Furthermore, the present specification discloses a method for enhancing apheresis by inducing cell mobilization in a subject, comprising the step of administering a CXCR4 inhibitor and G-CSF to a subject in the absence of a beta-adrenergic receptor inhibitor. Additionally, the present specification discloses a method for enhancing apheresis by mobilizing cells in a subject, comprising the step of administering a CXCR4 inhibitor and G-CSF to a subject in the absence of a beta-adrenergic receptor inhibitor. In some embodiments, administration of a combination of a CXCR4 inhibitor and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor alone. In some embodiments, administration of a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor alone. In some embodiments, administration of a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor alone. In some embodiments, administration of a combination of GPC-100 (burixafor, or TG-0054) and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by AMD3100 (plerixafor, or mozobil) and G-CSF. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by a CXCR4 inhibitor alone is 1.1 to 1.2 times, 1.2 to 1.3 times, 1.3 to 1.4 times, 1.4 to 1.5 times, 1.5 to 1.6 times, 1.6 to 1.7 times, 1.7 to 1.8 times, 1.8 to 1.9 times, 1.9 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 4 times, 4 to 5 times, 5 to 10 times, or 10 times or more. In some embodiments, the increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone is 5%-10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%-120% more, 120%-140% more, 140%-160% more, 160%-180% more, 180%-200% more, 200%-250% more, or 250%-300% more, 300%–400% more, 400%–500% more, 500%–750% more, 750%–1000% more, or 1000% or more. In some embodiments, the increased amount of cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) analysis. In some embodiments, the increased amount of cell mobilization or apheresis is measured by flow cytometry. In some embodiments, flow cytometry is (Lin. - Sca1 + c-Kit + ) is performed on LSK cells. In some embodiments, the increased amount of cell mobilization or component collection is measured by colony-forming unit (CFU) analysis.

[0084] Additional information regarding ADRB2 evaluated to form a heteromer with CXCR4 in this specification is described in detail below:

[0085] ADRB2 – The beta-2 adrenergic receptor (β2 adrenergic receptor), also known as ADRB2, is a transmembrane beta-adrenergic receptor that interacts with epinephrine (ligand synonym: adrenaline), a hormone and neurotransmitter that mediates physiological responses such as smooth muscle relaxation and bronchodilation through downstream L-type calcium channel interactions (Gregorio et al., 2017). ADRB2 functions in the muscular system, such as smooth muscle relaxation, motor nerve endings, and glycogenolysis, as well as in the circulatory system, such as myocardial contraction and increased cardiac output. In a normal eye, beta-2 stimulation by salbutamol increases intraocular pressure through the network. In the digestive system, ADRB2 induces glycogenolysis and gluconeogenesis in the liver and insulin secretion from the pancreas (Fitzpatrick, 2004).

[0086] ADRB2 signaling in cardiomyocytes is regulated by interaction with CXCR4 (LaRocca et al., 2010). Norepinephrine attenuates CXCR4 expression and the corresponding invasion of MDA-MB-231 breast cancer cells through ADRB2 (Wang et al., 2015a). ADRB2 is expressed in several cancers, such as pancreatic cancer, prostate cancer (Braadland et al., 2014; Xu et al., 2017), renal cancer, and breast cancer (Choy et al., 2016).

[0087] Alternative methods for detecting heteromer formation include, but are not limited to: immunostaining (Bushlin et al., 2012; Decaillot et al., 2008); immunoelectron microscopy; BRET (Pfleger and Eidne, 2006); time-resolved FRET analysis; in situ hybridization; FRET (Lohse et al., 2012); BRET, FRET, BiFC, bimolecular luminescence complementation, enzyme fragmentation analysis, and beta-arrestin recruitment analysis using GPCR heteromer identification technology (GPCR-HIT, Dimerix Bioscience) with the Tango Tango GPCR analysis system (Thermo Fisher Scientific); PRESTO-Tango system (Kroeze et al., 2015); controlled secretion / aggregation technology (ARIAD Pharmaceuticals) (Hansen et al., 2009); Receptor selection and amplification technology (ACADIA Pharmaceuticals) (Hansen et al., 2009); DimerScreen (Cara Therapeutics); dimer / interacting protein translocation assay (Patobios); co-immunoprecipitation; GPCR internalization assay using surface enzyme-linked immunosorbent assay (ELISA) (Decaillot et al., 2008) or flow cytometry (Law et al., 2005); whole-cell phosphorylation assay (Pfeiffer et al., 2002); and proximity ligation assay (PLA) (Frederick et al., 2015).

[0088] Alternative methods for detecting changes in pharmacological properties, signaling properties, and / or trafficking properties in cells expressing both CXCR4 and GPCRx include, but are not limited to: radioligand binding assays (Bushlin et al., 2012; Pfeiffer et al., 2002); cell surface biotinylation and immunoblotting; immunostaining (Bushlin et al., 2012; Decaillot et al., 2008); immunoelectron microscopy; [35S]GTPγS binding assays (Bushlin et al., 2012); calcium imaging or assays using dyes such as Fura 2-acetomethoxy ester (Molecular Probes), Fluo-4 NW calcium dye (Thermo Fisher Scientific), or FLIPR5 dye (Molecular Devices); cAMP assays using a radioimmunoassay kit (Amersham Biosciences); AlphaScreen (PerkinElmer Life Sciences); Parameter Cyclic AMP assay (R&D Systems); femto cAMP kit (Cisbio); cAMP direct immunoassay kit (Calbiochem) or GloSensor cAMP assay (Promega); GTPase assay (Pello et al., 2008); PKA activation (Stefan et al., 2007); ERK1 / 2 and / or Akt / PKB phosphorylation assay (Callen et al., 2012); Src and STAT3 phosphorylation assay (Rios et al., 2006); reporter assays such as cAMP response factor (CRE); nuclear factor response factor (NFAT-RE) of activated T-cells; serum response factor (SRE); serum response factor response factor (SRF-RE); and NF-κB-response factor luciferase reporter assay; secreted alkaline phosphatase assay (Decaillot et al., 2011); measurement of inositol 1-phosphate production using TR-FRET or [3H]myo-inositol; RT-qPCR for measuring downstream target gene expression; and adenylyl cyclase activity (George et al., 2000); next-generation sequencing (NGS); and any other assay capable of detecting changes in receptor function as a result of receptor heterodimerization.

[0089] Phrases used in this specification " Protein-protein interaction inhibitors ", " PPI inhibitors "Or variations thereof refer to any molecule capable of interfering with protein-protein interactions. Unlike enzyme-substrate interactions involving well-defined binding pockets, protein-protein interactions are transient interactions or bindings between proteins over a relatively wide area and are often induced by electrostatic interactions, hydrophobic interactions, hydrogen bonding, and / or van der Waals forces. PPI inhibitors may include, but are not limited to, membrane-permeable peptides or lipids fused to peptide sequences that interfere with GPCR heteromer interfaces, e.g., the transmembrane helix, intracellular loop, or C-terminal tail of GPCRx. For example, a PPI inhibitor of the CXCR4-GPCRx heteromer may be a membrane-permeable peptide or cell-penetrating peptide (CPP) conjugated with a peptide targeting the CXCR4-GPCRx heteromer interface(s), or a cell-penetrating lipidation targeting the CXCR4-GPCRx heteromer interface(s) It can be a peptide.

[0090] For example, membrane-permeable peptides or cell-penetrating peptides include the following: TAT 48-60 and TAT 49-57HIV-1 TAT peptides such as; Penetratin such as pAntp (43-58); polyarginine (Rn such as R5 to R12); Diatos peptide vector 1047 (DPV1047, Vectocell®); MPG (HIV gp41 fused to the nuclear localization signal (NLS) of SV40 large T antigen); Pep-1 (tryptophan-rich cluster fused to the NLS of SV40 large T antigen); pVEC peptide (vascular endothelial cadherin); p14 alternate read frame (ARF) protein-based ARF (1-22); N-terminus of raw bovine prion protein BPrPr (1-28); model amphiphilic peptide (MAP); Transportan; Azurin-derived p28 peptide; Amphiphilic β-sheet peptides such as VT5; proline-rich CPPs such as Bac 7 (Bac1-24); hydrophobic CPPs such as C105Y derived from α1-antitrypsin; PFVYLI derived from synthetic C105Y; Pep-7 peptide (CHL8 peptide phage clone); and modified hydrophobic CPPs such as stapled peptides and prenylated peptides (Guidotti et al., 2017; Kristensen et al., 2016). Membrane-permeable peptides or cell-penetrating peptides may further include, for example, TAT-derived cell-penetrating peptides, signal sequence-based (e.g., NLS) cell-penetrating peptides, hydrophobic membrane potential sequence (MTS) peptides, and arginine-rich molecular transporters. Cell-penetrating lipid-forming peptides include, for example, pepducin, such as ICL1 / 2 / 3 and C-tailed short palmitoylated peptides [47, 48].

[0091] The peptide(s) targeting the CXCR4-GPCRx heteromer interface may be, for example, the transmembrane domain of CXCR4, the transmembrane domain of GPCRx, the intracellular loop of CXCR4, the intracellular loop of GPCRx, the C-terminal domain of CXCR4, or the C-terminal domain of GPCRx, the extracellular loop of CXCR4, the extracellular loop of GPCRx, the N-terminal region of CXCR4, or the N-terminal region of GPCRx.

[0092] The present specification describes a method for treating a subject in need. In some embodiments, the method includes the step of blocking CXCR4 in the subject. In some embodiments, a drug is administered while blocking beta-adrenergic receptors in the subject. In some embodiments, a drug is administered while blocking a CXCR4 inhibitor and beta-adrenergic receptors in the subject.

[0093] In some embodiments, blocking of CXCR4 is achieved through ionic interaction with Arg188 on CXCR4. In some embodiments, blocking of CXCR4 is achieved through hydrogen bonding with Gln200 on CXCR4. In some embodiments, blocking of CXCR4 is achieved through a combination of ionic interaction with Arg188 on CXCR4 and hydrogen bonding with Gln200 on CXCR4.

[0094] The present specification describes a method for preparing a subject for treatment. In some embodiments, the method includes the step of administering an effective amount of a CXCR4 inhibitor. In some embodiments, the drug is administered as a beta-adrenergic receptor inhibitor. In some embodiments, the drug is administered as a combination thereof.

[0095] The present specification describes a method for qualifying a subject for treatment. In some embodiments, the method includes the step of administering an effective amount of a CXCR4 inhibitor. In some embodiments, the drug is administered as a beta-adrenergic receptor inhibitor. In some embodiments, the drug is administered as a combination thereof.

[0096] The present specification describes a method for treating a subject in need. In some embodiments, the method comprises the step of administering an effective amount of a CXCR4 inhibitor. In some embodiments, the drug is administered as a beta-adrenergic receptor inhibitor. In some embodiments, the drug is administered as a combination thereof.

[0097] In some embodiments, treatment further comprises administering an effective amount of a beta-adrenergic receptor inhibitor, or G-CSF, or lenalidomide, or daratumumab, or Velcade, or dexamethasone, or cyclophosphamide, or bortezombil, or a combination thereof.

[0098] In some embodiments, the subject is pretreated by administration of an effective amount of a beta-adrenergic receptor inhibitor, or G-CSF, or lenalidomide, or daratumumab, or Velacade, or dexamethasone, or cyclophosphamide, or bortezomib, or a combination thereof.

[0099] In some embodiments, the treatment comprises an apheresis step; or a leukocyte apheresis step; or a peripheral blood collection step; or a cell transplantation step, a CAR-T therapy step; or one or more combinations thereof. In some embodiments, the cells are optionally stem cells. In some embodiments, the cells are optionally hematopoietic progenitor cells (HSCs). In some embodiments, the cells are optionally T cells, or a combination thereof.

[0100] In some embodiments, the method further includes the step of administering an effective amount of G-CSF.

[0101] In some embodiments, the subject exhibits or is diagnosed with symptoms of cancer, or a neurological disorder, or any combination thereof. In some embodiments, the cancer is selected from the group consisting of lymphoma, leukemia, and myeloma. In some embodiments, the subject exhibits or is diagnosed with symptoms of blood cancer. In some embodiments, the cancer is selected from the group consisting of non-Hodgkin lymphoma (NHL), Hodgkin's disease (HD) or Hodgkin lymphoma (HL), acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), multiple myeloma (MM), and Burkitt lymphoma. In some embodiments, the cancer is HD. In some embodiments, the cancer is NHL. In some embodiments, the cancer is AML. In some embodiments, the cancer is ALL. In some embodiments, the cancer is MM. In some embodiments, the nervous system disorder is selected from the group consisting of nervous system disorders, cardiac ischemia, myocardial infarction, diabetes mellitus, tissue repair, bone and cartilage diseases, autoimmune diseases, graft-versus-host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the subject is a human.

[0102] In some embodiments, the subject requires autologous stem cell transplantation (ASCT). In some embodiments, the treatment is autologous stem cell transplantation (ASCT).

[0103] In some embodiments, the CXCR4 inhibitor forms an ionic interaction with Arg188 on CXCR4. In some embodiments, the CXCR4 inhibitor forms a hydrogen bond with Gln200 on CXCR4. In some embodiments, the CXCR4 inhibitor forms an ionic interaction with Arg188 on CXCR4 and forms a hydrogen bond with Gln200 on CXCR4.

[0104] In some embodiments, the CXCR4 inhibitor has a higher binding affinity for CXCL12 than AMD3100. In some embodiments, the CXCR4 inhibitor has a lower inhibition constant (Ki) than AMD3100. In some embodiments, the CXCR4 inhibitor has an inhibition constant at least twice as low as AMD3100.

[0105] In some embodiments, the CXCR4 inhibitor is ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549, D-[Lys3]GHRP-6, FC122, FC131, GMI-1359, GSK812397, GSTNT21MP, Isothiourea-1a, Isothiourea-1t (IT1t), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[11C]methyl-AMD3465, POL6326, SDF-1 l-9[P2G] dimer, SDFl P2G, T134, T140, T22, TC 14012, GPC-100 (burixafor, or TG-0054), USL311, viral macrophage inflammation protein-II (vMIP-11), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixater, [99mTc]O2-AMD3100, [177Lu]pentixater, ulocuplumab (MDX1338 / BMS-936564), AD-214, LY2624587, PF-06747143, POL6326. The inhibitor is selected from the group consisting of MB1707, 508MC1 (compound 26), and derivatives and analogs thereof. In some embodiments, the inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixapor, or Mozobil), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, GPC-100 (Bulixapor, or TG-0054), PF-06747143, POL6326, and ulokuflumab (MDX1338 / BMS-936564), and derivatives and analogs thereof. In some embodiments, the inhibitor is ulocuplumab (MDX1338 / BMS-936564).In some embodiments, the inhibitor is AMD3100 (Plerixaphor, or Mozobil). In some embodiments, the inhibitor is BL8040. In some embodiments, the inhibitor is GPC-100 (Bulixaphor, or TG-0054).

[0106] In some embodiments, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor. In some embodiments, the inhibitor is selected from the group consisting of alprenorolol, atenolol, betaxolol, bufranolol, butoxamine, carazolol, carvedilol, CGP 12177, cycloprorolol, ICI 118551, ICYP, labetalol, levobetaxolol, levovunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, timolol, and derivatives and analogs thereof. In some embodiments, the inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551. In some embodiments, the inhibitor is propranolol.

[0107] In some embodiments, the CXCR4 inhibitor is GPC-100, or AMD3100, or BL8040, preferably GPC-100. In some embodiments, the beta-adrenergic receptor inhibitor is propranolol.

[0108] In some embodiments, the effective dose of the CXCR inhibitor is a pharmacologically effective dose with clinically acceptable serious side effects.

[0109] In some embodiments, the effective amount of the CXCR inhibitor is:

[0110] It is about 30 mg / kg to about 0.01 mg / kg, or

[0111] It is about 30 mg / kg to about 0.07 mg / kg, or

[0112] It is about 25 mg / kg to about 0.07 mg / kg, or

[0113] It is about 20 mg / kg to about 0.07 mg / kg, or

[0114] It is about 15 mg / kg to about 0.07 mg / kg, or

[0115] It is about 10 mg / kg to about 0.07 mg / kg, or

[0116] About 9 mg / kg to about 0.07 mg / kg, or

[0117] It is about 8 mg / kg to about 0.07 mg / kg, or

[0118] It is about 7 mg / kg to about 0.07 mg / kg, or

[0119] It is about 6 mg / kg to about 0.07 mg / kg, or

[0120] It is about 5 mg / kg to about 0.07 mg / kg, or

[0121] About 4.40 mg / kg to about 0.07 mg / kg, or

[0122] About 3.14 mg / kg to about 0.07 mg / kg, or

[0123] About 2.24 mg / kg to about 0.07 mg / kg, or

[0124] About 1.12 mg / kg to about 0.07 mg / kg, or

[0125] About 0.56 mg / kg to about 0.07 mg / kg, or

[0126] About 0.28 mg / kg to about 0.07 mg / kg, or

[0127] About 0.14 mg / kg to about 0.07 mg / kg, or

[0128] About 0.10 mg / kg to about 0.07 mg / kg, or

[0129] It is about 30 mg / kg to about 0.10 mg / kg, or

[0130] It is about 25 mg / kg to about 0.10 mg / kg, or

[0131] It is about 20 mg / kg to about 0.10 mg / kg, or

[0132] It is about 15 mg / kg to about 0.10 mg / kg, or

[0133] It is about 10 mg / kg to about 0.10 mg / kg, or

[0134] It is about 9 mg / kg to about 0.10 mg / kg, or

[0135] It is about 8 mg / kg to about 0.10 mg / kg, or

[0136] It is about 7 mg / kg to about 0.10 mg / kg, or

[0137] It is about 6 mg / kg to about 0.10 mg / kg, or

[0138] It is about 5 mg / kg to about 0.10 mg / kg, or

[0139] It is about 4.40 mg / kg to about 0.10 mg / kg, or

[0140] About 3.14 mg / kg to about 0.10 mg / kg, or

[0141] About 2.24 mg / kg to about 0.10 mg / kg, or

[0142] About 1.12 mg / kg to about 0.10 mg / kg, or

[0143] About 0.56 mg / kg to about 0.10 mg / kg, or

[0144] About 0.28 mg / kg to about 0.10 mg / kg, or

[0145] About 0.14 mg / kg to about 0.10 mg / kg or

[0146] It is about 4.40 mg / kg.

[0147] In some embodiments, the effective dose of the CXCR inhibitor is about 3.14 mg / kg. In some embodiments, the effective dose of the CXCR inhibitor is about 2.24 mg / kg. In some embodiments, the effective dose of the CXCR inhibitor is about 1.12 mg / kg. In some embodiments, the effective dose of the CXCR inhibitor is about 0.56 mg / kg. In some embodiments, the effective dose of the CXCR inhibitor is about 0.28 mg / kg. In some embodiments, the effective dose of the CXCR inhibitor is about 0.14 mg / kg. In some embodiments, the effective dose of the CXCR inhibitor is about 0.10 mg / kg. In some embodiments, the effective dose of the CXCR inhibitor is about 0.07 mg / kg.

[0148] In some embodiments, the CXCR4 inhibitor is administered via a pharmacologically acceptable route. In some embodiments, the CXCR4 inhibitor is administered orally, subcutaneously, or intravenously.

[0149] In some embodiments, the duration of administration of the CXCR4 inhibitor is about 1 minute to about 30 minutes, or about 5 minutes to about 20 minutes, or about 10 minutes to about 18 minutes. In some embodiments, the CXCR4 inhibitor is administered at about 15 minutes.

[0150] In some embodiments, the time of administration of the CXCR4 inhibitor is:

[0151] It is morning, or

[0152] It is about 7:00 AM to about 7 PM, or

[0153] It is around 7:30 AM to around 12:30 PM, or

[0154] It is about 7:30 AM to about 9:30 PM.

[0155] In some embodiments, the time of administration of the CXCR4 inhibitor is about 8:30 AM.

[0156] In some embodiments, the last dose of the CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is:

[0157] When apheresis and / or leukocyte apheresis treatment begins

[0158] Administered about 120 hours to about 30 minutes prior, or

[0159] Administered about 72 hours to about 30 minutes prior, or

[0160] Administered about 24 hours to about 30 minutes prior, or

[0161] Administered about 14 hours to about 30 minutes prior, or

[0162] Administered about 13 hours to about 30 minutes prior, or

[0163] Administered about 13 hours to about 30 minutes prior, or

[0164] Administered about 12 hours to about 30 minutes prior, or

[0165] Administered about 11 hours to about 30 minutes prior, or

[0166] Administered about 10 hours to about 30 minutes prior, or

[0167] Administered about 9 hours to about 30 minutes prior, or

[0168] Administered about 8 hours to about 30 minutes prior, or

[0169] Administered about 7 hours to about 30 minutes prior, or

[0170] Administered about 6 hours to about 30 minutes prior, or

[0171] Administered about 5 hours to about 30 minutes prior, or

[0172] Administered about 4 hours to about 30 minutes prior, or

[0173] Administered about 3 hours to about 30 minutes prior, or

[0174] Administered about 2 hours to about 30 minutes prior, or

[0175] Administered about 90 minutes to about 30 minutes prior, or

[0176] Administered about 60 minutes to about 30 minutes prior, or

[0177] It is administered about 45 minutes to about 30 minutes before.

[0178] In some embodiments, the last dose of the CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is:

[0179] When apheresis and / or leukocyte apheresis treatment begins

[0180] Administered about 11 hours prior, or

[0181] Administered about 10 hours prior, or

[0182] Administered about 9 hours prior, or

[0183] Administered about 8 hours prior, or

[0184] Administered about 7 hours prior, or

[0185] Administered about 6 hours prior, or

[0186] Administered about 5 hours prior, or

[0187] Administered about 4 hours prior, or

[0188] It is administered about 3 hours in advance.

[0189] In some embodiments, the last dose of a CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 2 hours before the apheresis and / or leukocyte apheresis treatment begins.

[0190] In some embodiments, the last dose of a CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 90 minutes before the apheresis and / or leukocyte apheresis treatment begins.

[0191] In some embodiments, the last dose of a CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 60 minutes before the apheresis and / or leukocyte apheresis treatment begins.

[0192] In some embodiments, the last dose of a CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 45 minutes before the apheresis and / or leukocyte apheresis treatment begins.

[0193] In some embodiments, the last dose of a CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 30 minutes before the apheresis and / or leukocyte apheresis treatment begins.

[0194] In some embodiments, the CXCR4 inhibitor is administered once, twice, or three times, four times, or more than four times. In some embodiments, the CXCR4 inhibitor is administered one or fewer times, two or fewer times, three or fewer times, four or fewer times, or five or fewer times. In some embodiments, the CXCR4 inhibitor is administered for one day, two days, three days, or four days, or more than four days. In some embodiments, the CXCR4 inhibitor is administered for one day or fewer, two or fewer days, three or fewer days, four or fewer days, or five or fewer days.

[0195] In some embodiments, the apheresis and / or leukocyte apheresis treatment comprises about 1 to about 5 sessions, preferably about 1 to 2 sessions, more preferably 1 session.

[0196] In some embodiments, the CXCR4 inhibitor is administered once daily. In some embodiments, the CXCR4 inhibitor is administered about 7 to 11 days after propranolol administration. In some embodiments, the CXCR4 inhibitor is administered about 9 to 11 days after propranolol administration. In some embodiments, the CXCR4 inhibitor is administered about 7 to 8 days after propranolol administration.

[0197] In some embodiments, the CXCR4 inhibitor is administered on the 7th day after administration of propranolol. In some embodiments, the CXCR4 inhibitor is administered on the 8th day after administration of propranolol. In some embodiments, a first dose of the CXCR4 inhibitor is administered. In some embodiments, the CXCR4 inhibitor is administered about 7 to about 11 days after administration of propranolol. In some embodiments, the CXCR4 inhibitor is administered about 7 to about 8 days after administration of propranolol.

[0198] In some embodiments, the CXCR4 inhibitor is administered on the 7th day after administration of propranolol. In some embodiments, the CXCR4 inhibitor is administered on the 8th day after administration of propranolol. In some embodiments, the CXCR4 inhibitor is administered about 7 to about 11 days after administration of propranolol. In some embodiments, the CXCR4 inhibitor is administered about 9 to about 11 days after administration of propranolol.

[0199] In some embodiments, a CXCR4 inhibitor is administered on the 9th day after administration of propranolol.

[0200] In some embodiments, a CXCR4 inhibitor is administered on the 10th day after administration of propranolol.

[0201] In some embodiments, a CXCR4 inhibitor is administered on the 11th day after administration of propranolol.

[0202] In some embodiments, about 0.1 mg / kg to about 4.4 mg / kg of a CXCR4 inhibitor is administered intravenously once daily for 1 day or up to 5 days, and preferably, the last dose of the CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 30 to about 60 minutes before the apheresis and / or leukocyte apheresis treatment begins.

[0203] In some embodiments, the effective amount of GPC-100 is:

[0204] It is about 30 mg / kg to about 0.07 mg / kg, or

[0205] It is about 25 mg / kg to about 0.07 mg / kg, or

[0206] It is about 20 mg / kg to about 0.07 mg / kg, or

[0207] It is about 15 mg / kg to about 0.07 mg / kg, or

[0208] It is about 10 mg / kg to about 0.07 mg / kg, or

[0209] About 9 mg / kg to about 0.07 mg / kg, or

[0210] It is about 8 mg / kg to about 0.07 mg / kg, or

[0211] It is about 7 mg / kg to about 0.07 mg / kg, or

[0212] It is about 6 mg / kg to about 0.07 mg / kg, or

[0213] It is about 5 mg / kg to about 0.07 mg / kg, or

[0214] About 4.40 mg / kg to about 0.07 mg / kg, or

[0215] About 3.14 mg / kg to about 0.07 mg / kg, or

[0216] About 2.24 mg / kg to about 0.07 mg / kg, or

[0217] About 1.12 mg / kg to about 0.07 mg / kg, or

[0218] About 0.56 mg / kg to about 0.07 mg / kg, or

[0219] About 0.28 mg / kg to about 0.07 mg / kg, or

[0220] About 0.14 mg / kg to about 0.07 mg / kg, or

[0221] About 0.10 mg / kg to about 0.07 mg / kg, or

[0222] It is about 30 mg / kg to about 0.10 mg / kg, or

[0223] It is about 25 mg / kg to about 0.10 mg / kg, or

[0224] It is about 20 mg / kg to about 0.10 mg / kg, or

[0225] It is about 15 mg / kg to about 0.10 mg / kg, or

[0226] It is about 10 mg / kg to about 0.10 mg / kg, or

[0227] It is about 9 mg / kg to about 0.10 mg / kg, or

[0228] It is about 8 mg / kg to about 0.10 mg / kg, or

[0229] It is about 7 mg / kg to about 0.10 mg / kg, or

[0230] It is about 6 mg / kg to about 0.10 mg / kg, or

[0231] It is about 5 mg / kg to about 0.10 mg / kg, or

[0232] It is about 4.40 mg / kg to about 0.10 mg / kg, or

[0233] About 3.14 mg / kg to about 0.10 mg / kg, or

[0234] About 2.24 mg / kg to about 0.10 mg / kg, or

[0235] About 1.12 mg / kg to about 0.10 mg / kg, or

[0236] About 0.56 mg / kg to about 0.10 mg / kg, or

[0237] About 0.28 mg / kg to about 0.10 mg / kg, or

[0238] It is about 0.14 mg / kg to about 0.10 mg / kg.

[0239] In some embodiments, the effective dose of GPC-100 is about 4.40 mg / kg. In some embodiments, the effective dose of GPC-100 is about 3.14 mg / kg. In some embodiments, the effective dose of GPC-100 is about 2.24 mg / kg. In some embodiments, the effective dose of GPC-100 is about 1.12 mg / kg. In some embodiments, the effective dose of GPC-100 is about 0.56 mg / kg. In some embodiments, the effective dose of GPC-100 is about 0.28 mg / kg. In some embodiments, the effective dose of GPC-100 is about 0.14 mg / kg. In some embodiments, the effective dose of GPC-100 is about 0.10 mg / kg. In some embodiments, the effective dose of GPC-100 is about 0.07 mg / kg.

[0240] In a specific embodiment, GPC-100 is administered via a pharmacologically acceptable route. In a specific embodiment, GPC-100 is administered orally. In a specific embodiment, GPC-100 is administered subcutaneously. In a specific embodiment, GPC-100 is administered intravenously.

[0241] In a specific embodiment, the administration period of GPC-100 is:

[0242] It is about 1 minute to about 30 minutes, or

[0243] It is about 5 minutes to about 20 minutes, or

[0244] It is about 10 minutes to about 18 minutes, or

[0245] It is about 15 minutes.

[0246] In a specific embodiment, the administration time of GPC-100 is:

[0247] It is morning, or

[0248] It is about 7:00 AM to about 7 PM, or

[0249] It is around 7:30 AM to around 12:30 PM, or

[0250] It is around 7:30 AM to around 9:30 PM, or

[0251] It is about 8:30 AM.

[0252] In a specific embodiment, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is:

[0253] Administered about 120 hours to about 30 minutes prior, or

[0254] Administered about 72 hours to about 30 minutes prior, or

[0255] Administered about 24 hours to about 30 minutes prior, or

[0256] Administered about 14 hours to about 30 minutes prior, or

[0257] Administered about 13 hours to about 30 minutes prior, or

[0258] Administered about 13 hours to about 30 minutes prior, or

[0259] Administered about 12 hours to about 30 minutes prior, or

[0260] Administered about 11 hours to about 30 minutes prior, or

[0261] Administered about 10 hours to about 30 minutes prior, or

[0262] Administered about 9 hours to about 30 minutes prior, or

[0263] Administered about 8 hours to about 30 minutes prior, or

[0264] Administered about 7 hours to about 30 minutes prior, or

[0265] Administered about 6 hours to about 30 minutes prior, or

[0266] Administered about 5 hours to about 30 minutes prior, or

[0267] Administered about 4 hours to about 30 minutes prior, or

[0268] Administered about 3 hours to about 30 minutes prior, or

[0269] Administered about 2 hours to about 30 minutes prior, or

[0270] Administered about 90 minutes to about 30 minutes prior, or

[0271] Administered about 60 minutes to about 30 minutes prior, or

[0272] It is administered about 45 minutes to about 30 minutes before.

[0273] In a specific embodiment, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is:

[0274] When apheresis and / or leukocyte apheresis treatment begins

[0275] Administered about 11 hours prior, or

[0276] Administered about 10 hours prior, or

[0277] Administered about 9 hours prior, or

[0278] Administered about 8 hours prior, or

[0279] Administered about 7 hours prior, or

[0280] Administered about 6 hours prior, or

[0281] Administered about 5 hours prior, or

[0282] Administered about 4 hours prior, or

[0283] It is administered about 3 hours in advance.

[0284] In a specific embodiment, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered approximately 2 hours before the apheresis and / or leukocyte apheresis treatment begins. In a specific embodiment, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered approximately 90 minutes before the apheresis and / or leukocyte apheresis treatment begins. In a specific embodiment, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered approximately 60 minutes before the apheresis and / or leukocyte apheresis treatment begins. In a specific embodiment, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered approximately 45 minutes before the apheresis and / or leukocyte apheresis treatment begins. In a specific embodiment, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered about 30 minutes before the apheresis and / or leukocyte apheresis treatment begins.

[0285] In a specific embodiment, GPC-100 is administered once, twice, or three times, four times, or more than four times. In a specific embodiment, GPC-100 is administered one or fewer times, two or fewer times, three or fewer times, four or fewer times, or five or fewer times. In a specific embodiment, GPC-100 is administered for one day, two days, three days, or four days, or more than four days. In a specific embodiment, GPC-100 is administered for one or fewer days, two or fewer days, three or fewer days, four or fewer days, or five or fewer days.

[0286] In a specific embodiment, the apheresis and / or leukocyte apheresis treatment comprises about 1 to about 5 sessions, preferably about 1 to 2 sessions, more preferably 1 session.

[0287] In a specific embodiment, GPC-100 is administered once a day.

[0288] In a specific implementation example, the GPC-100 is:

[0289] It is administered about 7 to about 11 days after the administration of propranolol, or

[0290] It is administered about 9 to 11 days after the administration of propranolol, or

[0291] Administered about 7 to 8 days after administration of propranolol; or

[0292] Administered on the 7th day after administration of propranolol, or

[0293] It is administered on the 8th day after the administration of propranolol.

[0294] In a specific embodiment, the first capacity of the GPC-100 is:

[0295] It is administered about 7 to about 11 days after the administration of propranolol, or

[0296] It is administered about 7 to 8 days after the administration of propranolol, or

[0297] Administered on the 7th day after administration of propranolol, or

[0298] It is administered on the 8th day after the administration of propranolol.

[0299] In a specific embodiment, the second capacity of the GPC-100 is:

[0300] It is administered about 7 to about 11 days after the administration of propranolol, or

[0301] It is administered about 9 to 11 days after the administration of propranolol, or

[0302] Administered on the 9th day after administration of propranolol, or

[0303] Administered on the 10th day after administration of propranolol, or

[0304] It is administered on the 11th day after the administration of propranolol.

[0305] In some embodiments, about 0.1 mg / kg to about 4.4 mg / kg of GPC-100 is administered intravenously once daily for 1 day or up to 5 days, and preferably, the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered about 30 minutes to about 60 minutes before the apheresis and / or leukocyte apheresis treatment begins.

[0306] In some implementations, the effective amount of AMD3100 is:

[0307] It is about 10 μg / kg to about 480 μg / kg, or

[0308] About 40 μg / kg to about 480 μg / kg, or

[0309] It is about 80 μg / kg to about 480 μg / kg.

[0310] In some embodiments, the effective amount of AMD3100 is about 40 μg / kg to about 480 μg / kg and is administered subcutaneously. In some embodiments, the effective amount of AMD3100 is about 80 μg / kg to about 480 μg / kg and is administered subcutaneously.

[0311] In some embodiments, the effective amount of AMD3100 is about 40 μg / kg and is administered subcutaneously. In some embodiments, the effective amount of AMD3100 is about 80 μg / kg and is administered subcutaneously. In some embodiments, the effective amount of AMD3100 is about 480 μg / kg and is administered subcutaneously.

[0312] In some embodiments, the effective amount of AMD3100 is about 10 μg / kg to about 80 μg / kg and is administered intravenously. In some embodiments, the effective amount of AMD3100 is about 20 μg / kg to about 80 μg / kg and is administered intravenously. In some embodiments, the effective amount of AMD3100 is about 40 μg / kg to about 80 μg / kg and is administered intravenously.

[0313] In some embodiments, the effective amount of AMD3100 is about 10 μg / kg and is administered intravenously. In some embodiments, the effective amount of AMD3100 is about 20 μg / kg and is administered intravenously. In some embodiments, the effective amount of AMD3100 is about 40 μg / kg and is administered intravenously. In some embodiments, the effective amount of AMD3100 is about 80 μg / kg and is administered intravenously.

[0314] In a specific embodiment, AMD3100 is administered via a pharmacologically acceptable route. In a specific embodiment, AMD3100 is administered subcutaneously. In a specific embodiment, AMD3100 is administered intravenously.

[0315] In a specific embodiment, the last dose of the AMD3100 inhibitor prior to the start of apheresis and / or the start of leukocyte apheresis is:

[0316] When apheresis and / or leukocyte apheresis treatment begins

[0317] Administered about 120 hours to about 10 hours prior, or

[0318] Administered about 72 hours to about 10 hours prior, or

[0319] Administered about 24 hours to about 10 hours prior, or

[0320] Administered about 14 to 10 hours prior, or

[0321] Administered about 13 to 10 hours prior, or

[0322] Administered about 13 to 10 hours prior, or

[0323] Administered about 12 to 10 hours prior, or

[0324] Administered about 11 to 10 hours prior, or

[0325] Administered about 12 hours prior, or

[0326] Administered about 11 hours prior, or

[0327] It is administered about 10 hours in advance.

[0328] In a specific embodiment, AMD3100 is administered once a day. In a specific embodiment, AMD3100 is administered once, twice, three times, four times, or more than four times. In a specific embodiment, AMD3100 is administered five times or fewer, four times or fewer, three times or fewer, two times or fewer, or one time or fewer.

[0329] In some embodiments, AMD3100 is administered intravenously or subcutaneously, preferably subcutaneously, at a dose of about 10 μg / kg to about 480 μg / kg mg / kg once daily for 1 day or up to 5 days, and preferably the last dose of AMD3100 prior to apheresis and / or leukocyte apheresis is administered at least about 10 hours before the apheresis and / or leukocyte apheresis treatment begins, or preferably about 12 hours to about 10 hours before the apheresis and / or leukocyte apheresis treatment begins.

[0330] In some embodiments, the effective amount of BL8040 is:

[0331] About 0.5 mg / kg to about 2.0 mg / kg, or

[0332] About 0.75 mg / kg to about 2.0 mg / kg, or

[0333] It is about 1.0 mg / kg to about 2.0 mg / kg, or

[0334] About 1.25 mg / kg to about 2.0 mg / kg, or

[0335] It is about 1.5 mg / kg to about 2.0 mg / kg.

[0336] In some embodiments, the effective amount of BL8040 is about 0.5 mg / kg. In some embodiments, the effective amount of BL8040 is about 0.75 mg / kg. In some embodiments, the effective amount of BL8040 is about 1 mg / kg. In some embodiments, the effective amount of BL8040 is about 1.5 mg / kg.

[0337] In a specific embodiment, BL8040 is administered via a pharmacologically acceptable route. In a specific embodiment, BL8040 is administered subcutaneously. In a specific embodiment, BL8040 is administered intravenously.

[0338] In a specific embodiment, the last dose of BL8040 prior to the start of apheresis and / or the start of leukocyte apheresis is:

[0339] When apheresis and / or leukocyte apheresis treatment begins

[0340] Administered about 120 hours to about 10 hours prior, or

[0341] Administered about 72 hours to about 10 hours prior, or

[0342] Administered about 24 hours to about 10 hours prior, or

[0343] Administered about 14 to 10 hours prior, or

[0344] Administered about 13 to 10 hours prior, or

[0345] Administered about 13 to 10 hours prior, or

[0346] Administered about 12 to 10 hours prior, or

[0347] Administered about 11 to 10 hours prior, or

[0348] Administered about 12 hours prior, or

[0349] Administered about 11 hours prior, or

[0350] It is administered about 10 hours in advance.

[0351] In a specific embodiment, BL8040 is administered once a day. In a specific embodiment, BL8040 is administered 1, 2, 3, 4, or more than 4 times. In a specific embodiment, BL8040 is administered 5 or fewer times, 4 or fewer times, 3 or fewer times, 2 or fewer times, or 1 or fewer times.

[0352] In a specific embodiment, BL8040 is administered subcutaneously at about 1.5 mg / kg once daily for 1 day or up to 5 days, and preferably the last dose of BL8040 prior to apheresis and / or leukocyte apheresis is administered at least about 10 hours before the apheresis and / or leukocyte apheresis treatment begins, preferably about 12 hours to about 10 hours before the apheresis and / or leukocyte apheresis treatment begins.

[0353] In a specific embodiment, the effective dose of the beta-adrenergic receptor inhibitor is a pharmacologically effective dose having clinically acceptable serious side effects.

[0354] In a specific embodiment, the effective amount of the beta-adrenergic receptor inhibitor is:

[0355] About 10 mg to about 480 mg per day, or

[0356] About 20 mg to about 480 mg per day, or

[0357] About 30 mg to about 480 mg per day, or

[0358] About 40 mg to about 480 mg per day, or

[0359] About 50 mg to about 480 mg per day, or

[0360] About 60 mg to about 480 mg per day, or

[0361] About 70 mg to about 480 mg per day, or

[0362] About 80 mg to about 480 mg per day, or

[0363] About 120 mg to about 480 mg per day, or

[0364] About 160 mg to about 480 mg per day, or

[0365] About 200 mg to about 480 mg per day, or

[0366] About 240 mg to about 480 mg per day, or

[0367] About 280 mg to about 480 mg per day, or

[0368] About 320 mg to about 480 mg per day, or

[0369] About 360 mg to about 480 mg per day, or

[0370] About 400 mg to about 480 mg per day, or

[0371] About 440 mg to about 480 mg per day, or

[0372] About 10 mg to about 480 mg per day, or

[0373] About 10 mg to about 450 mg per day, or

[0374] It is about 10 mg to about 420 mg per day, or

[0375] About 10 mg to about 390 mg per day, or

[0376] About 10 mg to about 360 mg per day, or

[0377] About 10 mg to about 330 mg per day, or

[0378] About 10 mg to about 300 mg per day, or

[0379] About 10 mg to about 270 mg per day, or

[0380] About 10 mg to about 240 mg per day, or

[0381] About 10 mg to about 210 mg per day, or

[0382] It is about 10 mg to about 180 mg per day, or

[0383] About 10 mg to about 150 mg per day, or

[0384] It is about 10 mg to about 120 mg per day, or

[0385] It is about 10 mg to about 90 mg per day, or

[0386] It is about 10 mg to about 60 mg per day, or

[0387] It is about 10 mg to about 30 mg per day, or

[0388] It is about 10 mg to about 90 mg per day, or

[0389] It is about 10 mg to about 60 mg per day, or

[0390] It is about 10 mg to about 30 mg per day, or

[0391] About 20 mg to about 480 mg per day, or

[0392] About 20 mg to about 450 mg per day, or

[0393] About 20 mg to about 440 mg per day, or

[0394] About 20 mg to about 410 mg per day, or

[0395] About 20 mg to about 400 mg per day, or

[0396] About 20 mg to about 370 mg per day, or

[0397] About 20 mg to about 360 mg per day, or

[0398] About 20 mg to about 330 mg per day, or

[0399] About 20 mg to about 320 mg per day, or

[0400] About 20 mg to about 290 mg per day, or

[0401] About 20 mg to about 280 mg per day, or

[0402] About 20 mg to about 250 mg per day, or

[0403] About 20 mg to about 240 mg per day, or

[0404] About 20 mg to about 210 mg per day, or

[0405] About 20 mg to about 200 mg per day, or

[0406] About 20 mg to about 170 mg per day, or

[0407] About 20 mg to about 160 mg per day, or

[0408] About 20 mg to about 130 mg per day, or

[0409] About 20 mg to about 120 mg per day, or

[0410] It is about 20 mg to about 90 mg per day, or

[0411] It is about 20 mg to about 80 mg per day, or

[0412] It is about 20 mg to about 50 mg per day, or

[0413] It is about 20 mg to about 40 mg per day, or

[0414] About 20 mg to about 160 mg per day, or

[0415] About 20 mg to about 130 mg per day, or

[0416] About 20 mg to about 120 mg per day, or

[0417] About 20 mg to about 100 mg per day, or

[0418] It is about 20 mg to about 80 mg per day, or

[0419] It is about 20 mg to about 70 mg per day, or

[0420] It is about 20 mg to about 40 mg per day.

[0421] In some embodiments, the effective dose of the beta-adrenergic receptor inhibitor is about 20 mg per day. In some embodiments, the effective dose of the beta-adrenergic receptor inhibitor is about 40 mg per day. In some embodiments, the effective dose of the beta-adrenergic receptor inhibitor is about 60 mg per day. In some embodiments, the effective dose of the beta-adrenergic receptor inhibitor is about 80 mg per day, and / or

[0422] In a specific embodiment, the beta-adrenergic receptor inhibitor is administered via a pharmacologically acceptable route. In a specific embodiment, the beta-adrenergic receptor inhibitor is administered orally.

[0423] In a specific embodiment, a beta-adrenergic receptor inhibitor is administered once a day, or twice a day, or three times a day, or four times a day.

[0424] In a specific embodiment, the beta-adrenergic receptor inhibitor is:

[0425] Administered at intervals of approximately 7 to 12 hours, or

[0426] Administered at intervals of approximately 12 hours, or

[0427] Administered at intervals of approximately 11 hours, or

[0428] Administered at intervals of approximately 10 hours, or

[0429] Administered at intervals of approximately 9 hours, or

[0430] Administered at intervals of approximately 8 hours, or

[0431] Administered at intervals of approximately 7 hours, or

[0432] Administered at intervals of approximately 6 hours, or

[0433] Administered at intervals of approximately 5 hours, or

[0434] Administered at intervals of approximately 4 hours, or

[0435] It is administered at intervals of about 3 hours.

[0436] In a specific embodiment, a beta-adrenergic receptor inhibitor is administered orally at a dose of about 30 mg twice a day at intervals of about 7 to 12 hours.

[0437] In a specific embodiment, the effective dose of the beta-adrenergic receptor inhibitor is a pharmacologically effective dose having clinically acceptable serious side effects.

[0438] In a specific embodiment, the effective amount of the ADRB2 inhibitor is:

[0439] About 10 mg to about 480 mg per day, or

[0440] About 20 mg to about 480 mg per day, or

[0441] About 30 mg to about 480 mg per day, or

[0442] About 40 mg to about 480 mg per day, or

[0443] About 50 mg to about 480 mg per day, or

[0444] About 60 mg to about 480 mg per day, or

[0445] About 70 mg to about 480 mg per day, or

[0446] About 80 mg to about 480 mg per day, or

[0447] About 120 mg to about 480 mg per day, or

[0448] About 160 mg to about 480 mg per day, or

[0449] About 200 mg to about 480 mg per day, or

[0450] About 240 mg to about 480 mg per day, or

[0451] About 280 mg to about 480 mg per day, or

[0452] About 320 mg to about 480 mg per day, or

[0453] About 360 mg to about 480 mg per day, or

[0454] About 400 mg to about 480 mg per day, or

[0455] About 440 mg to about 480 mg per day, or

[0456] About 10 mg to about 480 mg per day, or

[0457] About 10 mg to about 450 mg per day, or

[0458] It is about 10 mg to about 420 mg per day, or

[0459] About 10 mg to about 390 mg per day, or

[0460] About 10 mg to about 360 mg per day, or

[0461] About 10 mg to about 330 mg per day, or

[0462] About 10 mg to about 300 mg per day, or

[0463] About 10 mg to about 270 mg per day, or

[0464] About 10 mg to about 240 mg per day, or

[0465] About 10 mg to about 210 mg per day, or

[0466] It is about 10 mg to about 180 mg per day, or

[0467] About 10 mg to about 150 mg per day, or

[0468] It is about 10 mg to about 120 mg per day, or

[0469] It is about 10 mg to about 90 mg per day, or

[0470] It is about 10 mg to about 60 mg per day, or

[0471] It is about 10 mg to about 30 mg per day, or

[0472] It is about 10 mg to about 90 mg per day, or

[0473] It is about 10 mg to about 60 mg per day, or

[0474] It is about 10 mg to about 30 mg per day, or

[0475] About 20 mg to about 480 mg per day, or

[0476] About 20 mg to about 450 mg per day, or

[0477] About 20 mg to about 440 mg per day, or

[0478] About 20 mg to about 410 mg per day, or

[0479] About 20 mg to about 400 mg per day, or

[0480] About 20 mg to about 370 mg per day, or

[0481] About 20 mg to about 360 mg per day, or

[0482] About 20 mg to about 330 mg per day, or

[0483] About 20 mg to about 320 mg per day, or

[0484] About 20 mg to about 290 mg per day, or

[0485] About 20 mg to about 280 mg per day, or

[0486] About 20 mg to about 250 mg per day, or

[0487] About 20 mg to about 240 mg per day, or

[0488] About 20 mg to about 210 mg per day, or

[0489] About 20 mg to about 200 mg per day, or

[0490] About 20 mg to about 170 mg per day, or

[0491] About 20 mg to about 160 mg per day, or

[0492] About 20 mg to about 130 mg per day, or

[0493] About 20 mg to about 120 mg per day, or

[0494] It is about 20 mg to about 90 mg per day, or

[0495] It is about 20 mg to about 80 mg per day, or

[0496] It is about 20 mg to about 50 mg per day, or

[0497] It is about 20 mg to about 40 mg per day, or

[0498] About 20 mg to about 160 mg per day, or

[0499] About 20 mg to about 130 mg per day, or

[0500] About 20 mg to about 120 mg per day, or

[0501] About 20 mg to about 100 mg per day, or

[0502] It is about 20 mg to about 80 mg per day, or

[0503] It is about 20 mg to about 70 mg per day, or

[0504] It is about 20 mg to about 40 mg per day, or

[0505] About 20 mg per day, or

[0506] About 40 mg per day, or

[0507] Approximately 60 mg per day, or

[0508] It is about 80 mg per day.

[0509] In a specific embodiment, the ADRB2 inhibitor is administered via a pharmacologically acceptable route. In a specific embodiment, the ADRB2 inhibitor is administered orally.

[0510] In a specific embodiment, the ADRB2 inhibitor is administered once a day, or twice a day, or three times a day, or four times a day.

[0511] In a specific embodiment, the ADRB2 inhibitor is:

[0512] It is administered at intervals of approximately 7 to 12 hours, or

[0513] Administered at intervals of approximately 12 hours, or

[0514] Administered at intervals of approximately 11 hours, or

[0515] Administered at intervals of approximately 10 hours, or

[0516] Administered at intervals of approximately 9 hours, or

[0517] Administered at intervals of approximately 8 hours, or

[0518] Administered at intervals of approximately 7 hours, or

[0519] Administered at intervals of approximately 6 hours, or

[0520] Administered at intervals of approximately 5 hours, or

[0521] Administered at intervals of approximately 4 hours, or

[0522] It is administered at intervals of about 3 hours.

[0523] In a specific embodiment, an ADRB2 inhibitor is administered orally at a dose of about 30 mg twice a day at intervals of about 7 to 12 hours.

[0524] In a specific embodiment, the effective amount of propranolol is:

[0525] About 10 mg to about 480 mg per day, or

[0526] About 20 mg to about 480 mg per day, or

[0527] About 30 mg to about 480 mg per day, or

[0528] About 40 mg to about 480 mg per day, or

[0529] About 50 mg to about 480 mg per day, or

[0530] About 60 mg to about 480 mg per day, or

[0531] About 70 mg to about 480 mg per day, or

[0532] About 80 mg to about 480 mg per day, or

[0533] About 120 mg to about 480 mg per day, or

[0534] About 160 mg to about 480 mg per day, or

[0535] About 200 mg to about 480 mg per day, or

[0536] About 240 mg to about 480 mg per day, or

[0537] About 280 mg to about 480 mg per day, or

[0538] About 320 mg to about 480 mg per day, or

[0539] About 360 mg to about 480 mg per day, or

[0540] About 400 mg to about 480 mg per day, or

[0541] About 440 mg to about 480 mg per day, or

[0542] About 10 mg to about 480 mg per day, or

[0543] About 10 mg to about 450 mg per day, or

[0544] It is about 10 mg to about 420 mg per day, or

[0545] About 10 mg to about 390 mg per day, or

[0546] About 10 mg to about 360 mg per day, or

[0547] About 10 mg to about 330 mg per day, or

[0548] About 10 mg to about 300 mg per day, or

[0549] About 10 mg to about 270 mg per day, or

[0550] About 10 mg to about 240 mg per day, or

[0551] About 10 mg to about 210 mg per day, or

[0552] It is about 10 mg to about 180 mg per day, or

[0553] About 10 mg to about 150 mg per day, or

[0554] It is about 10 mg to about 120 mg per day, or

[0555] It is about 10 mg to about 90 mg per day, or

[0556] It is about 10 mg to about 60 mg per day, or

[0557] It is about 10 mg to about 30 mg per day, or

[0558] It is about 10 mg to about 90 mg per day, or

[0559] It is about 10 mg to about 60 mg per day, or

[0560] It is about 10 mg to about 30 mg per day, or

[0561] About 20 mg to about 480 mg per day, or

[0562] About 20 mg to about 450 mg per day, or

[0563] About 20 mg to about 440 mg per day, or

[0564] About 20 mg to about 410 mg per day, or

[0565] About 20 mg to about 400 mg per day, or

[0566] About 20 mg to about 370 mg per day, or

[0567] About 20 mg to about 360 mg per day, or

[0568] About 20 mg to about 330 mg per day, or

[0569] About 20 mg to about 320 mg per day, or

[0570] About 20 mg to about 290 mg per day, or

[0571] About 20 mg to about 280 mg per day, or

[0572] About 20 mg to about 250 mg per day, or

[0573] About 20 mg to about 240 mg per day, or

[0574] About 20 mg to about 210 mg per day, or

[0575] About 20 mg to about 200 mg per day, or

[0576] About 20 mg to about 170 mg per day, or

[0577] About 20 mg to about 160 mg per day, or

[0578] About 20 mg to about 130 mg per day, or

[0579] About 20 mg to about 120 mg per day, or

[0580] It is about 20 mg to about 90 mg per day, or

[0581] It is about 20 mg to about 80 mg per day, or

[0582] It is about 20 mg to about 50 mg per day, or

[0583] It is about 20 mg to about 40 mg per day, or

[0584] About 20 mg to about 160 mg per day, or

[0585] About 20 mg to about 130 mg per day, or

[0586] About 20 mg to about 120 mg per day, or

[0587] About 20 mg to about 100 mg per day, or

[0588] It is about 20 mg to about 80 mg per day, or

[0589] It is about 20 mg to about 70 mg per day, or

[0590] It is about 20 mg to about 40 mg per day.

[0591] In a specific embodiment, the effective amount of propranolol is about 20 mg per day. In a specific embodiment, the effective amount of propranolol is about 40 mg per day. In a specific embodiment, the effective amount of propranolol is about 60 mg per day. In a specific embodiment, the effective amount of propranolol is about 80 mg per day.

[0592] In a specific embodiment, propranolol is administered via a pharmacologically acceptable route.

[0593] In a specific embodiment, propranolol is administered orally.

[0594] In a specific embodiment, propranolol is administered once a day, or twice a day, or three times a day, or four times a day.

[0595] In a specific embodiment, propranolol is:

[0596] It is administered at intervals of approximately 7 to 12 hours, or

[0597] Administered at intervals of approximately 12 hours, or

[0598] Administered at intervals of approximately 11 hours, or

[0599] Administered at intervals of approximately 10 hours, or

[0600] Administered at intervals of approximately 9 hours, or

[0601] Administered at intervals of approximately 8 hours, or

[0602] Administered at intervals of approximately 7 hours, or

[0603] Administered at intervals of approximately 6 hours, or

[0604] Administered at intervals of approximately 5 hours, or

[0605] Administered at intervals of approximately 4 hours, or

[0606] It is administered at intervals of about 3 hours.

[0607] In a specific embodiment, about 30 mg of propranolol is administered orally twice a day at intervals of about 7 to 12 hours.

[0608] In a specific embodiment, the effective amount of G-CSF is:

[0609] Administered intravenously at about 1 µg / kg to about 70 µg / kg twice daily, or

[0610] Administered by subcutaneous injection at a dose of about 1 µg / kg to about 40 µg / kg per day, or

[0611] It is about 3 ug / kg to about 40 ug / kg per day by continuous subcutaneous injection.

[0612] In a specific embodiment, the effective amount of G-CSF is about 10 ug / kg per day to about 40 ug / kg per day. In a specific embodiment, the effective amount of G-CSF is about 10 ug / kg per day.

[0613] In a specific embodiment, G-CSF is administered via a pharmacologically acceptable route. In a specific embodiment, G-CSF is administered intravenously. In a specific embodiment, G-CSF is administered subcutaneously. In a specific embodiment, G-CSF is administered continuously subcutaneously.

[0614] In a specific embodiment, the last dose of the G-CSF inhibitor prior to the start of apheresis and / or the start of leukocyte apheresis is:

[0615] When apheresis and / or leukocyte apheresis treatment begins

[0616] Administered about 120 hours to about 10 hours prior, or

[0617] Administered about 72 hours to about 10 hours prior, or

[0618] Administered about 24 hours to about 10 hours prior, or

[0619] Administered about 14 to 10 hours prior, or

[0620] Administered about 13 to 10 hours prior, or

[0621] Administered about 13 to 10 hours prior, or

[0622] Administered about 12 to 10 hours prior, or

[0623] Administered about 11 to 10 hours prior, or

[0624] Administered about 12 hours prior, or

[0625] Administered about 11 hours prior, or

[0626] It is administered about 10 hours in advance.

[0627] In a specific embodiment, G-CSF is administered once a day. In a specific embodiment, G-CSF is administered 1, 2, 3, 4, 5, 6, 7, 8, or more than 8 times. In a specific embodiment, G-CSF is administered 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, or 3 times or less, 2 times or less, or 1 time or less.

[0628] In a specific embodiment, G-CSF is administered by intravenous injection, subcutaneous injection, or serial subcutaneous injection at a rate of about 10 µg / kg per day for about 4 days, preferably at least about 10 hours prior to apheresis and / or leukocyte apheresis.

[0629] In some embodiments, CD34 to the object + A drug that interferes with cell mobilization is administered. In some embodiments, the drug is lenalidomide. In some embodiments, the drug is administered according to the drug label. In some embodiments, the drug is administered at a dose of about 8 mg / kg to about 16 mg / kg, preferably about 16 mg / kg. In some embodiments, the drug is administered via a pharmacologically acceptable route. In some embodiments, the pharmacologically acceptable route is intravenous. In some embodiments, the drug is administered for about 3 to about 4 cycles.

[0630] In some embodiments, the drug is daratumumab. In some embodiments, the drug daratumumab is administered according to the drug label. In some embodiments, the drug is administered at a dose of about 8 mg / kg to about 16 mg / kg, preferably about 16 mg / kg. In some embodiments, the drug is administered via a pharmacologically acceptable route. In some embodiments, the drug is administered intravenously. In some embodiments, the drug is administered for about 3 to about 4 cycles.

[0631] In some embodiments, the method does not cause adverse events in the subject during or after treatment. In some embodiments, the method does not cause severe adverse events in the subject during or after treatment. In some embodiments, the method does not cause adverse events of greater than grade 1 or 2. In some embodiments, adverse events are limited to a list consisting of bone pain, hypocalcemia, diarrhea, dry mouth, nausea, hypokalemia, peripheral edema, paranesthesia, etc., if caused.

[0632] The present specification describes the use of GPC-100 for the manufacture of a drug for the treatment of cancer, or neurological disorders, or any combination thereof.

[0633] The novel CXCR4 antagonist GPC-100 is effective for cell mobilization, either alone or in combination with other agents.

[0634] As demonstrated in Examples 1 to 6, the novel CXCR4 antagonist GPC-100 is an effective hematopoietic cell mobilization agent and has a functional interaction between CXCR4 and β2AR, which supports the combination of GPC-100 and propranolol as a new strategy for HSPC mobilization.

[0635] GPC-100 demonstrated efficacy comparable to AMD3100 in interfering with CXCL12-mediated calcium signaling, cell migration, and β-arrestin recruitment to CXCR4. However, in vivo, GPC-100 was a superior mobilizer compared to AMD3100. Competitive binding analysis also revealed that GPC-100 possesses superior binding to CXCR4 compared to AMD3100 and has additional binding modes for enhanced interaction with CXCR4 compared to AMD3100.

[0636] Regardless of whether it is activating or inhibiting, the functional outcome of ligand binding depends on the structural characteristics of the ligand-receptor complex. This affects signaling molecules such as G-proteins or arrestin-recruiting GPCR kinases involved in the intracellular cavity of GPCRs (Gurevich Frontiers in Pharmacology 2019). For example, Jorgensen et al. showed that AMD11070, another small molecule CXCR4 antagonist, had a higher binding affinity for CXCR4 and a more potent efficacy in blocking CXCL12-induced migration than AMD3100, but AMD3100 was a superior mobilizer. This was attributed to biased signaling of β-arrestin recruitment by AMD3100 and differences in binding modes

[73] .

[0637] CXCR4 and β2AR are known to form heteromer complexes with other GPCRs [63, 74-77]. Receptor heteromerization can lead to changes in ligand binding affinity, G protein-coupling, efficacy, receptor trafficking, or β-arrestin interactions

[78] . Successfully investigating GPCR heteromerization in endogenous, non-transformed native cells has been a difficult task due to factors such as low protein abundance and a lack of detection methods

[62] .

[0638] Using highly sensitive and specific assays such as PLA, this technique is the first to demonstrate that the two receptors are close enough to form heteromers or at least allow receptor crosstalk using cancer cells endogenously expressing CXCR4 and β2AR

[79] . The detection of CXCR4 and β2AR in cancer cells is further supported by the use of highly sensitive and accurate pulsed interleave excitation fluorescence cross-correlation spectroscopy to detect co-localization suggesting close physical interaction between CXCR4 and ADRB2 (Sukhtankar, 2023b). Co-activation of CXCR4 and β2AR led to enhanced calcium flux and increased recruitment of β-arrestin to CXCR4. This indicates that the proximity of the two receptors has downstream functional consequences mediated by both G-protein-dependent and independent pathways, emphasizing the importance of GPCR heteromers as drug targets.

[0639] It is known that CXCR4-mediated calcium signaling regulates cell migration and survival

[22] . Recent studies suggest that β2AR may utilize a non-canonical, G-protein-independent mechanism to induce calcium flux

[80] . Therefore, as observed in this technique, the increased recruitment of β-arrestin to CXCR4 induced by epinephrine may lead to a mega-complex

[81] linking CXCR4-β2AR-β-arrestin to the calcium activation mechanism.

[0640] Although the role of calcium flux in HSC biology is still being elucidated

[82] , calcium flux has been shown to activate the calcineurin / NFAT (nuclear factor of activated T cells) pathway and downstream gene reprogramming, leading to changes in HSC maintenance and differentiation

[83] . Therefore, an increase in calcium flux due to CXCR4-β2AR co-activation may be involved in HSC recruitment. Collectively, this technique suggests the potential for signaling crosstalk between CXCR4 and β2AR, opening up opportunities for new therapeutic approaches.

[0641] HSPC mobilization is associated with a concomitant increase in circulating WBCs [84-87]. In mice, propranolol significantly enhanced GPC-100-induced WBC mobilization. Previously, β2AR and CXCR4 were shown to interact physically and functionally to enhance lymphocyte retention within lymph nodes

[26] . Given that both receptors are expressed on HSCs, similar interactions may also play a role in HSC retention by CXCR4 within the BM. However, due to technical limitations, these physical interactions in HSCs have not yet been demonstrated. Knight et al. discussed that propranolol can not only promote HSC growth and differentiation but also improve BM cellularity [38, 88-90]. In particular, propranolol treatment [indicated] CD34 in MM patients + It induced upregulation of HSC-like gene signatures and diverted cell differentiation away from myeloid bias, which can lead to superior engraftment in ASCT.

[0642] This technique demonstrates that propranolol improves GPC-100-induced HSPC recruitment by a factor of two. Considering that GPC-100 can clinically recruit up to 12-fold HSPCs with PB

[23] , the increase resulting from the addition of propranolol to GPC-100 can provide sufficient HSPC harvesting. This suggests the possibility of recruiting HSCs without the use of G-CSF. This is an important finding because excluding G-CSF from treatment can reduce the risk of moderate to severe side effects of G-CSF, such as severe bone pain and, rarely, splenic rupture

[16] .

[0643] Patients with MM not only have a high symptom burden and poor health-related quality of life

[91] , but also have a history of treatment that negatively affects G-CSF mobilization. An alternative approach using oral beta-blockers such as propranolol eliminates the toxicity caused by repeated mobilization attempts, as well as the risks and burdens associated with daily subcutaneous G-CSF injections. Collectively, these studies suggest the benefits of adding propranolol to HSC mobilization therapy.

[0644] The addition of propranolol to G-CSF and GPC-100 (triple combination) resulted in a significant increase in viable functional HSPCs compared to G-CSF and AMD3100. These cells were functionally able to differentiate into myeloid and erythroid multipotent progenitor cells and also expressed primitive markers known for the organ reorganization ability of HSCs. CXCR4 and β2AR are expressed in most immune cells

[28] . Propranolol significantly increased GPC-100-induced lymphocyte mobilization.

[0645] In summary, the findings of this technology suggest therapeutic co-targeting of the CXCR4 and β2AR pathways and support the addition of propranolol to GPC-100 for HSC mobilization for ASCT in MM patients. The triple combination of GPC-100, propranolol, and G-CSF can potentially target patient populations for whom other mobilization therapies have failed. Propranolol has been proven to be a safe, accessible, and affordable option to supplement mobilization therapies to obtain higher stem cell yields with fewer apheresis sessions, thereby reducing the economic burden on patients and the healthcare system. This is currently being tested in a two-arm Phase 2 clinical trial (NCT05561751)

[92] including the GPC-100 and propranolol group and the GPC-100, propranolol, and G-CSF group.

[0646] Clinical studies show that combination therapy with GPC-100 and G-CSF in patients with multiple myeloma (MM), non-Hodgkin lymphoma (NHL), and / or Hodgkin's disease (HD) is generally well tolerated and effective for cell mobilization.

[0647] The need to mobilize cells from bone marrow into the PB for collection, particularly CD34+ HSCs for stem cell transplantation and T cells for CAR-T therapy, has increased over the past few years. HSC transplantation can extend progression-free survival in various hematological cancers, the most common of which are MM, NHL, and HD (Flomenberg 2005). Additionally, gene editing therapies for diseases such as sickle cell disease and β-thalassemia also utilize HSC transplantation. CAR-T therapy has also been observed to induce effective and sustained responses in hematological cancers. The clinical efficacy of these therapies depends on whether a sufficient number of cells are injected for functional reconstitution. 2. Mobilized PB is the preferred source of HSCs compared to bone marrow aspiration due to improved engraftment success rates, increased safety and convenience, shortened hospital stay, and lower overall cost (Beyer 1995, Devine 2004, Hartmann 1997, Smith 1997, To 1997, Vose 2002). To achieve this, a mobilization agent must be administered to the patient before cell collection in a procedure called leukocyte apheresis (Liles 2005).

[0648] Two mobilization agents approved by the FDA are AMD3100 (plerixafor, or mozovil) and BL8040 (moticaportide), which are antagonists of human CXCR4. For some disease indications, these mobilization agents are used in combination with G-CSF, which has an orthogonal mechanism of action and is contraindicated in some autoimmune diseases such as sickle cell disease. Both AMD3100 and BL8040 have similar efficacy and must be administered 10 to 14 hours prior to leukocyte apheresis for HSC transplantation in hematological malignancies. Typically, patients receive these agents on the evening before leukocyte apheresis and must begin the procedure the following morning. This adds complexity and burden to patients already undergoing a complex procedure. It has been argued that the administration of BL8040 may be more flexible because CD34+ cells remain in the periphery for a longer period. However, the impact of this on the stemness of these stem cells and their redistribution to other organs has not been sufficiently studied. One indication that such a state of stem cells may not be entirely harmless is the dyspnea experienced by some patients upon G-CSF administration due to lung tissue infiltration by mobilized stem cells. Although combination therapy with AMD3100 and G-CSF or BL8040 and G-CSF is an improvement over G-CSF monotherapy, mobilization is still inhibited in approximately 15–35% of MM patients (DiPersio 2009b). Considering these factors, alternative mobilization agents with differentiated characteristics are needed.

[0649] GPC-100 is a potent and selective antagonist of CXCR4. In nonclinical studies, GPC-100 was shown to possess a distinct binding mode and higher affinity for CXCR4 compared to AMD3100. In mice, GPC-100 was also shown to mobilize more WBCs to the PB compared to AMD3100 (Sukhtankar, 2023a). Nonclinical data suggest that GPC-100 exhibits synergistic effects when co-administered with G-CSF. In a previous Phase 1 study, administration of GPC-100 at a single dose of 0.10 to 4.40 mg / mL resulted in a safe and well-tolerated formulation in 64 healthy volunteers (Sukhtankar 2023b).

[0650] As shown in Examples 10 to 12 describing a randomized, open-label, double-blind, placebo-controlled, multicenter, phase 2 clinical trial (NCT02104427), combination therapy of GPC-100 and G-CSF in patients with multiple myeloma (MM), non-Hodgkin lymphoma (NHL) and / or Hodgkin's disease (HD) is generally well tolerated and effective for cell mobilization.

[0651] G-CSF therapy can be used for patients receiving HSC transplants (Neupogen label), but when used alone, low CD34 after each component apheresis + Generates cell numbers, and optimal CD34 after a median of 4 days of apheresis + It was shown to achieve cell numbers (DiPersio 2009a). In a Phase 2 study, combination therapy with GPC-100 and G-CSF was well tolerated, and optimal CD34 was achieved within 2 days (average 1.4 days) via apheresis. + It resulted in achieving a cell count of ≥5.0×10 6 CD34 +The primary endpoint of collecting cells / kg was met in 11 out of 12 (92%) MM (n=8) or NHL or HD (n=3) patients undergoing apheresis within 2 days. Among the 9 MM patients, 8 patients (89%) had ≥6.0×10 within an average of one leukocyte apheresis session. 6 CD34 + Cells / kg were mobilized. The only MM patient who did not meet the primary endpoint had been treated with lenalidomide, which is known to reduce stem cell yield in patients receiving HSC transplants (Cashen 2007). Total CD34 collected after GPC-100 and G-CSF treatment + The cell / kg count was higher in MM patients compared to NHL and HD patients, and the average cumulative cell count was 9.00×10⁶, respectively. 6 and 5.68×10 6 CD34 + It was cells / kg. The ability of GPC-100 and G-CSF to induce greater mobilization in MM patients compared to NHL and HD patients is consistent with existing data after administration of AMD3100 combined with G-CSF (H (bel 2012). When AMD3100 and G-CSF were used, 59% of NHL patients ≥5.0×10 within 4 or fewer leukocyte apheresis sessions (median 3 sessions). 6 CD34 cells / kg + ...was mobilized, and 72% of MM patients had ≥6.0×10 within 2 or fewer leukocyte apheresis sessions (median 1 session). 6 CD34 cells / kg + mobilized (AMD3100 or Pleric Safor product label 2007).

[0652] It is important to emphasize that, in contrast to AMD3100 or motic saportide (10-11 hours) and G-CSF, the effects of GPC-100 and G-CSF on stem cell mobilization were achieved within 1-2 hours after treatment (Crees et al). This difference may be due to the higher binding affinity of GPC-100 to CXCR4 (Sukhtankar, 2023). The rapid effects of GPC-100 will enable a simplified treatment regimen that reduces hospitalization resources and minimizes patient burden.

[0653] Although cell mobilization via a combination of G-CSF therapy and the CXCR4 antagonists GPC-100 or AMD3100 is improved compared to G-CSF monotherapy, HSC mobilization may be negatively affected in MM patients treated with daratumumab (Hulin 2021) or lenalidomide (Popat 2009). Furthermore, due to the negative side effects of G-CSF, such as bone pain, and contraindications in sickle cell disease and autoimmune diseases, it is important to find other options for stem cell mobilization. The literature provides evidence regarding the involvement of β-adrenergic receptor signaling and the use of beta-blockers in hematological malignancies (Hwa 2017). Beta-blockers have been shown to improve survival outcomes in MM patients (Hwa 2021).

[0654] To further optimize ASCT using GPC-100, recent preclinical studies have demonstrated enhanced stem cell mobilization with the use of GPC-100 combined with the beta-blocker propranolol (Sukhtankar, 2023). In mice, the combination of GPC-100 and propranolol was a potent mobilizer for WBCs, which was further enhanced by the addition of G-CSF. This effect was also replicated in mouse stem cell mobilization as measured by analysis of lineage-negative Sca1-positive and c-kit-positive stem cell populations and by colony-forming unit analysis. The use of the triple combination of GPC-100, propranolol, and G-CSF is predicted to be best in class and can target patient populations for whom other therapies have failed. However, mobilization via the dual combination of GPC-100 and propranolol may serve as a therapeutic option when G-CSF is intolerant.

[0655] In conclusion, the results of this Phase 2 study show that the combined administration of GPC-100 and G-CSF produces an optimal number of CD34s within 1–2 hours and after one or two apheresis sessions in patients with MM, NHL, or HD. + It was proven that cells can be mobilized.

[0656] Clinical studies demonstrate that combination therapy using GPC-100, propranolol, and G-CSF is safe and well-tolerated in multiple myeloma (MM) patients undergoing ASCT.

[0657] ASCT plays a central role in the treatment of MM. However, poor HPC mobilization occurs in 15-25% of patients. Newer therapies such as daratumumab can also have a negative impact on mobilization, which supports the need for alternative mobilization therapies.

[0658] GPC-100 is a potent and selective small molecule antagonist of CXCR4. Previous clinical trials of GPC-100 alone or in combination with G-CSF have demonstrated the safe and effective mobilization of stem cells. GPC-100 features faster mobilization kinetics compared to currently FDA-approved CXCR4 inhibitors and enables same-day administration and leukocyte apheresis.

[0659] Propranolol is a non-selective beta-blocker and has been clinically demonstrated to inhibit molecular risk markers in MM patients undergoing ASCT. Other studies have also shown that propranolol can improve cellularity within the bone marrow and reduce pro-tumorigenicity. This U.S.-only, open-label, multicenter Phase 2 clinical study (NCT05561751) evaluates the safety and migration efficacy of MM patients undergoing ASCT when treated with a combination of GPC-100, propranolol, and G-CSF.

[0660] As shown in Examples 13 to 14 describing the U.S.-only, open-label, multicenter Phase 2 clinical study (NCT05561751), combination therapy with GPC-100, propranolol, and G-CSF in MM patients undergoing ASCT is generally safe and tolerable, and CD34 + It efficiently mobilizes cells. Furthermore, this study demonstrates that a mobilization effect exists despite the negative mobilization effects of daratumumab.

[0661] Examples

[0662] The reagents for the embodiments of the present technology are provided in Table 3.

[0663]

[0664] Example 1: GPC-100 exhibits a distinct binding mode and higher CXCR4 binding affinity compared to AMD3100.

[0665] The chemical composition of GPC-100 shows a distinct difference from that of AMD3100 (Fig. 1a). Previous studies using various CXCR4 antagonists have identified major and minor pockets of interaction within the CXCR4 orthosteric binding site

[49] . To understand the inactive structure of CXCR4 at the molecular level, the CXCR4 antagonists GPC-100 and AMD3100 were studied using in silico docking experiments.

[0666] Simply put, Schr using the OPLS3e force field The Dinger Small-Molecular Drug Discovery Suite was used for the molecular modeling procedure. A human CXCR4 model (PDB: 3ODU) complexed with the small molecule antagonist IT1t was imported and prepared using the Protein Preparation Wizard workflow. AMD3100 and GPC-100 were prepared using LigPrep in default settings. A total of 25 and 18 different conformations were generated for AMD3100 and GPC-100, respectively. The standard workflow for ligand docking in XP (ultra-precision) settings for AMD3100 and GPC-100 to the CXCR4 structure was followed

[49] . For GPC-100, induced fit docking was also performed as previously described

[50] . To ensure proper docking of the binding pocket, residues Y116 / D171 / Y255 / E288 were selected as the center point. The trim option was selected to avoid steric collision with the large side chain Arg188. For further analysis, the top 20 poses were saved for GPC-100 with an energy window of 30 kcal / mol.

[0667] As previously described regarding the GPC-100, Schr Dinger's inductive fitting algorithm was used [49, 50]. Residues with previously verified secondary amines, namely Asp97 and Asp262

[51] , contributed to the interaction of GPC-100 and AMD3100 with CXCR4. However, the phosphate group of GPC-100, which is absent in AMD3100, forms ionic interactions with Arg188 and hydrogen bonds with Gln200 (Fig. 1b). These energetically favorable interactions suggest an enhanced anchor point for GPC-100, which is absent in AMD3100.

[0668] Next, to test whether structural differences could lead to differences in ligand binding, TagLite ® The CisBio system was used for competitive binding analysis. After plating CXCR4 terbium-labeled cells, 5 μL of GPC-100 or AMD3100 was added, followed by the addition of 5 μL of CXCL12, a 20 nM red fluorescently labeled CXCR4 ligand that competes with the CXCR4 inhibitor for CXCR4 binding. After incubation at room temperature for 3 hours, the plates were read using a PerkinElmer EnVision plate reader. The inhibition constant (Ki) for each compound was determined by plotting the compound concentration against the normalized HTRF ratio using the nonlinear regression competitive binding "single-site-fit Ki equation." The HTRF ratio was calculated as the 665 nm / 620 nm emission ratio proportional to the amount of CXCL12 bound to CXCR4.

[0669] The dose-response curves of GPC-100 or AMD3100 demonstrated that the antagonists competitively inhibit the binding of CXCL12 to CXCR4. As shown in Figure 2, GPC-100 has a higher affinity for CXCR4 than AMD3100. The inhibition constant (Ki) of GPC-100 is approximately 1.6 nM, whereas the Ki of AMD3100 is approximately 40 nM.

[0670] Example 2: Analysis of the pharmacological properties of GPC-100 demonstrates efficacy in inhibiting the CXCR4 / CXCL12 axis at a level comparable to or superior to that of AMD3100.

[0671] cell culture MDA-MB-231, Namalwa and multiple myeloma cells MM.1S from ATCC (American Type Culture Collection), and acute myeloid leukemia cells U937 from the Korean Cell Line Bank (Seoul, Korea) were cultured in ATCC modified RPMI 1640 (Gibco, A1049101) supplemented with 10% fetal bovine serum (FBS) and 100 U / ml penicillin-streptomycin.

[0672] Calcium flux analysis : MDA-MB-231 cells transfected with an adenovirus encoding CXCR4 for endogenous expression of CXCR4 and β2AR or for overexpression were used in calcium flux. Cells were cultured as previously described 2 days prior to the analysis run

[60] and placed in 8x10 96-well black clear-bottom microplates. 4 Cells were seeded into wells. The next day, cells were stained with Cal-520 AM (AAT Bioquest) at 37°C for 2 hours, treated with an antagonist for 30 minutes, and then stimulated with an agonist. Intracellular calcium flux was measured using a Flexstation 3 microplate reader (Molecular Devices).

[0673] Movement AnalysisU937 or MM.1S cells were treated with GPC-100 or AMD3100 for 30 minutes, and 100 μl of cell suspension was added to a Transwell insert with 8-μm pores. The Transwell insert containing the 100 μl cell suspension was placed in the bottom chamber, taking care to avoid air bubbles. 800 μl of serum-free medium containing 0.5% BSA was added to the bottom chamber of the Transwell at the specified concentration (Figs. 5a-5b). Cells were allowed to migrate toward CXCL12 for 4 hours (MM.1S) or 24 hours (U937) at 37°C in the presence or absence of an antagonist. Migration of U937 cells was measured using PrestoBlue cell viability reagent (Invitrogen). Fluorescence intensity (560 nm / 590 nm excitation / emission) was detected using a Varioskan LUX multimode microplate reader.

[0674] PRESTO-Tango Analysis : The CXCR4-dV2-Tango plasmid was constructed by removing the V2 tail of the V2 vasopressin receptor through Age I digestion using the CXCR4-Tango plasmid (Addgene plasmid #66262)

[61] . HTLA cells were cultured as previously described.

[61] 8 x 10⁶ HTLA cells were placed in a 6-well plate. 5Canine cells were seeded per well. The following day, cells were transfected with CXCR4-dV2 Tango and pcDNA3.1-ADRB2 using Lipofectamine 3000 (Thermo Fisher Scientific). After 48 hours of incubation, the transfected cells were transferred to 96-well white-bottom plates and incubated for 4 hours under serum-starvation conditions. Cells were pretreated with an antagonist for 30 minutes, followed by stimulation with an agonist for 18 hours. On the 5th day, cells were lysed with Steady-Glo solution (Promega) at room temperature for 15 minutes, and luminescence was measured using a Varioskan LUX multimode microplate reader.

[0675] In summary, the above analysis demonstrates the potent effect of GPC-100 on CXCR4 signaling. First, calcium flux analysis in CXCR4-overexpressing MDA-MB-231 cells (Table 4) showed a potent calcium flux response upon stimulation with CXCL12. Dose-response titration results of GPC-100 and AMD3100 revealed dose-dependent inhibition of CXCL12-induced calcium flux, and IC50 50 The values ​​were 30 nM and 35 nM, respectively (Fig. 3a). β-arrestin recruitment to CXCR4, known to occur upon CXCL12 binding, was also measured [52-54]. In the PRESTO-TANGO β-arrestin assay

[55] , both GPC-100 and AMD3100 inhibited CXCL12-mediated β-arrestin recruitment to CXCR4, with IC50 values ​​of 207 nM and 172 nM, respectively (Fig. 3b). Cell migration, directly associated with stem cell recruitment, is a key phenotypic response upon CXCR4 activation [56-58]. For this assay, U937 and MM.1S cells with high endogenous CXCR4 expression were used (Fig. 4)

[59] , which are sub-nanomolar EC 50They showed a potent migration response to CXCL12 with values ​​(Fig. 5). In U937 cells, both GPC-100 and AMD3100 efficiently blocked CXCL12-mediated migration, and IC 50 The values ​​were 48 nM and 42 nM, respectively (Fig. 3c). In MM.1S cells, CXCL12-induced migration was blocked by GPC-100 at significantly lower concentrations than AMD3100 (compare the GPC-100 IC50 value at 28 nM with the AMD3100 IC50 value at 80 nM (Fig. 3d)).

[0676] Example 3: Endogenously expressed CXCR4 and β 2 AR exists in close proximity in the model cell system.

[0677] Previous studies on cardiomyocytes and lymphocytes suggest that CXCR4 and β2AR form heteromers. Here, we investigated the heteromerization of CXCR4 and β2AR in the native state where the receptors are endogenously expressed. To test this, a proximity ligation assay (PLA) capable of detecting protein-protein interactions was applied using endogenous and unengineered receptors

[62] . Since PLA detects amplified DNA as dots, it can generate strong signals from only a small number of interacting molecules, enabling sensitive and specific analysis. Namalwa and MDA-MB-231 cells were used as model cancer cell lines selected based on receptor mRNA transcription levels, surface expression, and cell type diversity (Table 4, Fig. 5, and Figs. 6a-6b).

[0678] We observed positive PLA signals (more than 5 PLA events per cell) in Namalwa cells using primary antibodies specific to the intracellular domains of CXCR4 and β2AR, indicating stable proximity of endogenous CXCR4 and β2AR receptors (Fig. 7a). To confirm the specificity of receptor proximity, CXCR4 knockout was performed on Namalwa cells, and a significant decrease in surface CXCR4 expression was observed (Figs. 8a-8b). PLA signals were significantly reduced in CXCR4 knockout Namalwa cells compared to parent cells (Fig. 7a). While these results were encouraging, detection by PLA was difficult due to the relatively low abundance of β2AR in Namalwa cells.

[0679] Next, we investigated MDA-MB-231 cells expressing higher levels of β2AR mRNA (Table 4). In this cell system, PLA generated about 6 events per cell (Fig. 7b). The specificity of PLA was confirmed in the β2AR knockout of MDA-MB-231 cells (Fig. 7b) (Figs. 8c and 8d), suggesting that parental CXCR4 and β2AR exist in close proximity in the native cell environment.

[0680] RT-qPCR for CXCR4 and ADRB2 Total RNA was isolated using the rNeasy Mini Kit (Qiagen, Valencia, CA). One-strand cDNA synthesis and RT-qPCR were performed using the High-Volume cDNA Reverse Transcription Kit and Power SYBR Green PCR Master Mix (Applied Biosystems, Foster City, CA), respectively. The primer sequences are as follows:

[0681] CXCR4 : 5'-CCACCATCTACTCCATCATCTTC-3' and

[0682] 5'-ACTTGTCCGTCATGCTTCTC-3';

[0683] ADRB2 : 5'-CTCTTCCATCGTGTCCTTCTAC-3' and

[0684] 5'-AATCTTCTGGAGCTGCCTTT-3'E; and

[0685] ACTB (β-actin) : 5'-GGACTTCGAGCAAGAGATGG-3' and

[0686] 5'-AGCACTGTGTTGGCGTACAG-3'.

[0687] PCR conditions : 50°C for 2 minutes, followed by 95°C for 10 minutes, then 95°C for 15 seconds and 60°C for 1 minute, after 40 cycles, 95°C for 15 seconds, 60°C for 1 minute, and 95°C for 1 second for melting curve analysis using QuantStudio 3 instrument (Applied Biosystems). The expression of CXCR4 and ADRB2 mRNA was normalized to the expression of ACTB mRNA.

[0688] Creation of knockout cells :

[0689] Non-targeted single guide RNA (sgControl: 5′-ACGGAGGCTAAGCGTCGCAA-3′),

[0690] CXCR4-targeting sgRNA (sgCXCR4: 5′-ACTTACACTGATCCCCTCCA-3′), and ADRB2-targeting sgRNA (sgADRB2: 5'-CGTCTGCAGACGCTCGAACT-3') were cloned into pLentiCRISPRv2, respectively. Lentiviruses were produced and gene targeting was performed as previously described

[63] . Namalwa-CXCR4 KO cells were used in a pool form after puromycin selection. MDA-MB-231-ADRB2 KO clones were selected using the limiting dilution method after puromycin selection. Surface expression of CXCR4 and ADRB2 in parent cells and KO cells was detected using flow cytometry with anti-CXCR4 antibodies, anti-β2AR antibodies, or isotype control antibodies.

[0691]

[0692] Proximal Ligation Analysis (PLA) MDA-MB-231 (mother and ADRB2 knockout) and Namalwa (mother and CXCR4 knockout) cells were fixed to microscope slides and treated with lambda protein phosphatase (New England Biolabs) at 37°C for 1 hour. The NaveniFlex MR kit (Navinci) was used for blocking, primary antibody incubation, and PLA. Recombinant rabbit anti-CXCR4 antibody (clone UMB2, Abcam) and mouse anti-human β2AR antibody (clone E-3, Santa Cruz) were used as primary antibodies, and rabbit IgG and mouse IgG2b isotypes (Abcam) were used as negative controls. Slides were mounted using VECTASHIELD Antifade Mounting Medium containing DAPI and imaged using an IN Cell Analyzer 2500HS system (Molecular Devices).

[0693] Statistical analysis Data analysis was performed using GraphPad Prism, and all data are presented as mean ± SEM. Data comparisons between two administration conditions were conducted using the Mann-Whitney test, or, in the case of multiple dose conditions, one-way ANOVA combined with Turkey's test of multiple comparisons. For all tests, P < 0.05 was considered statistically significant. * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001, **** ≤ 0.0001.

[0694] Example 4: CXCR4 and β inhibited by GPC-100 and propranolol 2 Synergistic pharmacology upon co-activation of AR

[0695] The functional effects of CXCR4 and β2AR proximity were evaluated in downstream signaling assays using receptor co-activation and co-inhibition. β-arrestin acts as an intracellular scaffold mediating receptor desensitization, internalization, and G-protein-independent signaling [64-66]. HTLA cells overexpressing CXCR4-tango and β2AR were treated in two separate experiments with 0–200 nM CXCL12 and 1 μM epinephrine or 100 nM CXCL12 and 0–10 μM epinephrine. Under these conditions, CXCL12 alone induced β-arrestin recruitment to CXCR4 (Fig. 9a). Epinephrine alone also induced minimal but significant β-arrestin recruitment to CXCR4 (Fig. 9b). However, when these cells were co-treated with CXCL12 and epinephrine, a significant synergistic increase in β-arrestin recruitment was observed compared to treatment with CXCL12 alone (Fig. 9c, E upon co-treatment max 286 ± 12.20%), which was blocked by the combination of GPC-100 and propranolol (Fig. 9c).

[0696] When MDA-MB-231 cells endogenously expressing CXCR4 and β2AR (Fig. 10) were treated with CXCL12 or epinephrine alone, a minimal calcium flux response was induced. However, co-treatment with CXCL12 and epinephrine resulted in a synergistic increase in calcium flux (Figs. 11a and 11b). This was partially inhibited by single antagonist treatment using GPC-100, AMD3100, or propranolol (Fig. 11b). Complete blockade of calcium flux was achieved only by co-treatment with GPC-100 and propranolol. In contrast, co-treatment with AMD3100 and propranolol failed to significantly reduce the synergistically increased calcium flux. These results demonstrate that GPC-100 in the presence of propranolol is a more potent CXCR4 antagonist than AMD3100 in the presence of propranolol. In addition, these results suggest that the closely located CXCR4 and β2AR can be co-activated to functionally regulate downstream calcium signaling pathways, as well as influence receptor internalization mechanisms and potentially, scaffolds mediating G-protein-independent non-porous signaling pathways.

[0697] Example 5: GPC-100 was a significantly more effective mobilization agent than AMD3100, and combination therapy with GPC-100 and propranolol in vivo improves mobilization in mice.

[0698] The most effective dose for induced mobilization was evaluated for the CXCR4 antagonists GPC-100 and AMD3100. Numerous studies in mice suggest that peak mobilization by AMD3100 (5 mg / kg, SC) occurs at 1 hour [67-69]. Mouse PB WBC counts were determined 1 hour after administration of GPC-100 and AMD3100. The results showed that both GPC-100 and AMD3100 increased circulating WBCs compared to the vehicle (Fig. 12a). Importantly, among the antagonists, GPC-100 was significantly more effective than AMD3100 in increasing WBCs relative to the vehicle baseline; 1 hour after drug administration, GPC-100 showed a 4.6-fold increase, while AMD3100 showed a 3-fold increase (Fig. 12a).

[0699] The recruitment time course of GPC-100 and AMD3100 was also compared from 30 minutes to 6 hours after drug administration. Both antagonists showed peak recruitment at 2 hours in mice, but GPC-100-induced recruitment was significantly greater than that of AMD3100, and the recruitment effect of GPC-100 lasted longer than that of AMD3100 (Fig. 12b). Based on these time courses, PB was collected 2 hours after GPC-100 administration in subsequent studies.

[0700] animalThe study was conducted in a facility accredited by the Association for Accreditation of Laboratory Animal Care (AAALAC) and strictly adhered to the recommendations of the National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals. The protocol was approved by the Institutional Animal Care and Use Committee (IACUC) of Explora BioLabs (Protocol # EB17-010-141). C57BL / 6J mice (female, 6–9 weeks of age) obtained from Jackson Laboratories were acclimatized for one week prior to treatment. Mice were housed five per cage and had free access to food and water. Mice were randomized for each study to ensure that all treatment groups included similar age and body weight distributions. The test substances were administered as shown in Table 5.

[0701]

[0702] Sample CollectionMice were monitored daily, and body weight was measured prior to drug administration. Using body weight data, mice were randomized into groups using the randomization tool from Benchling Studies. Peripheral blood (PB) was collected via submental bleeding (non-terminal) or cardiac puncture (terminal). For terminal blood collection, an initial dose of 2.5–4% isoflurane was administered to mice in an induction chamber, followed by a maintenance dose of 2–3% via a nose cone. The surgical anesthesia phase was confirmed by applying a toe pinch to all four zones. Once blood collection from cardiac puncture was complete, mice were euthanized via cervical dislocation while still under the influence of isoflurane. Approximately 25 μL of blood was used for CBC analysis, and the remaining blood was processed for flow cytometry or colony-forming unit (CFU) analysis. For non-terminal submandibular blood sampling, the linguofacialis vein under the jaw was punctured using a 4 mm lancet. Blood was collected 1 or 2 hours after the administration of GPC-100 and AMD3100, and 14 hours after the administration of G-CSF. To track the time course of mobilization, blood was collected twice from each mouse. Submandibular blood sampling was performed at 0.5, 1, or 2 hours after administration, and cardiac puncture was performed at 3, 4, and 6 hours after administration. All doses of the therapeutic agents used in the study were well tolerated, and no clinical observations were noted during the procedure.

[0703] CBC analysis Blood samples were processed for complete blood count (CBC) analysis using an Abaxis VetScan HM5 blood analyzer that reports 18 parameters including WBC, lymphocytes, neutrophils, monocytes, platelets, hemoglobin, RBC, and morphology.

[0704] Flow cytometry analysis: LSK Cell (Lineage- Sca-1 + c-Kit + The recruitment of mouse HSCs characterized by )

[70] was determined in two separate studies. In both studies, mononuclear cells were isolated from PB after CBC analysis, and for the first study, they were stained with anti-lineage cocktail, c-Kit, and Sca-1 antibodies (Table 6). In the second study, HSCs with organ reconstitution ability (LT-HSC) (CD150 + or CD34 - CD150 or CD34 antibodies were added to determine LSK

[71] . Samples were acquired using a Cytek Aurora Spectrum Flow Cytometry System (Fremont, CA), and data were analyzed using CellEngine software. Gating was determined using an FMO control group. - c-Kit as a subgroup of cells + Sca-1 + The percentage of cells was used to determine the total number of LSK cells per μL of blood. In the second study, CD150 + or CD34 - The frequency of cells was measured as a subpopulation of parent LSK cells. The number of LSK cells in PB was normalized based on the WBC count obtained from the specific study.

[0705]

[0706] Colony Forming Unit (CFU) Analysis : 8x10 separated from PB after CBC analysis 5Dogs of monocytes were added to a tube containing semi-solid methylcellulose medium (StemCell Technologies) known to support erythrocytes and myeloid progenitor cells

[72] . After 7 days, colonies showing the appearance of granulocyte-monocyte progenitor cells (CFU-GM) and burst forming units erythroid (BFU-E) were formed and counted by a blind experimenter. Total CFU was calculated as the total number of CFU-GM and BFU-U colonies.

[0707] The CXCR4 antagonists GPC-100 and AMD3100, respectively, exhibited more pronounced mobilization in mice in the presence of the β2AR antagonist propranolol. To evaluate the effects of β2AR blockade in vivo, mice were administered propranolol. Propranolol doses were selected based on dose titrations (5–40 mg / kg, IP) when co-administered with GPC-100 (Fig. 13). Pretreatment with propranolol (20 mg / kg, IP) for 7 days (Fig. 14a) enhanced GPC-100-induced mobilization by 27% (Fig. 14b). Propranolol pretreatment also enhanced AMD3100-induced mobilization (Fig. 14b). Notably, propranolol improved lymphocyte mobilization in both antagonists but did not improve neutrophil migration (Figs. 15a and 15b). In additional experiments to determine HSPC mobilization, it was confirmed that 7 days of propranolol pretreatment enhanced LSK cell mobilization by GPC-100, resulting in a fourfold increase in circulating HSPC (Figs. 14c-14d).

[0708] Example 6: Triple combination therapy of G-CSF, GPC-100, and propranolol in vivo improves mobilization in mice

[0709] Mobilization studies from triple combinations of G-CSF, GPC-100 or AMD3100, and propranolol were compared to dual or monotherapy combinations under current ASCT standard treatment. The triple combination (Fig. 16a), as well as the combination of G-CSF and GPC-100, induced WBC mobilization 8.2-fold and 8.4-fold, respectively; this was significantly greater compared to the increased WBC counts caused by G-CSF alone (4.5-fold) or G-CSF and AMD3100 (6.6-fold) (Fig. 16b). Taken together, these results indicated that GPC-100 mobilized more cells than AMD3100 when administered alone or in combination with propranolol or G-CSF. Treatment with G-CSF and AMD3100 increased LSK cells in the PB 13-fold relative to the vehicle. In contrast, the combination of G-CSF and GPC-100 containing or not containing propranolol increased LSK cells by 20-fold and 24-fold, respectively (Fig. 16c, Fig. 17).

[0710] Next, the functional capacity of cells mobilized by the triple combination was compared in CFU analysis. CFU analysis was performed to measure the mobilized HSPCs based on their colony-forming ability in CFU-GM and BFU-E. The triple combination increased CFU 47-fold compared to the vehicle control, which compares to 35-fold and 27-fold increases for G-CSF and GPC-100 treatment and G-CSF and AMD3100 treatment, respectively, compared to the vehicle (Fig. 18a). LSK cells expressing CD150 or lacking CD34 are known to possess long-term remodeling ability (LT-HSC) (Purton Experimental Hematology 2022). In this technique, the triple combination compared CD34 - and CD150 +It significantly mobilized more LSK cells (Figs. 18b and 18c). This indicates that the addition of propranolol can improve the functional capacity of the mobilized cells. The patterns of LSK and CFU numbers across various drug combinations were consistent with the WBC numbers of the matched samples, which supports the use of WBC numbers as a surrogate marker for stem cell mobilization.

[0711] Example 7: Novel GPC-100 alone induced significantly more in vivo recruitment than AMD3100 and BL8040.

[0712] Time course of WBC subpopulation mobilization by GPC-100 (burixafor, or TG-0054), AMD3100 (plerixafor, or mozovil), and BL8040 (moticaportide), respectively, in mice. As shown in Figures 19a-19c, GPC-100-induced mobilization (not BL8040) was significantly greater than that induced by AMD3100. Peak neutrophil mobilization for GPC-100 occurred 2 hours after drug administration, whereas for BL8040 it occurred 4 hours later. This indicates greater mobilization of hematopoietic lineage cells. Subpopulation analysis for these cells will be performed for their potential utility in adoptive cell therapy.

[0713] The superiority of GPC-100 against BL8040 was enhanced particularly in the balb / c mouse strain. The balb / c mouse strain is primarily used for in vivo oncology studies. For mobilization studies, the C57 / BL6 strain (mentioned in other studies) is more commonly used due to the preferential G-CSF mobilization in this strain and the availability of co-genic mice. As shown in Figures 20a-20c, BL8040 is not efficiently mobilized in balb / c mice. balb / c produces more Th2-related cytokines with high IL4 and low IFNy, whereas C57 / BL6 produces more Th1-related cytokines with low IL4 and high IFNy. The Th2 profile is predominant in females. As used herein, " Th " refers to T helper cells, which additionally " Th1 " and " Th2 It is subdivided into "

[0714] Th1-dominant responses are involved in the pathogenesis of organ-specific autoimmune disorders, Crohn's disease, sarcoidosis, acute renal allograft rejection, and some unexplained recurrent miscarriages. In contrast, allergen-specific Th2 responses cause atopic disorders in genetically susceptible individuals. How Th1 and Th2 immune responses or Th1 / Th2 balance affect mobilization by CXCR4 inhibitors has not been investigated. It is unclear what these findings mean clinically.

[0715] Example 8: The combination of GPC-100 and propranolol improves the mobilization of myeloid cells (non-lymphocytes) in vivo.

[0716] As shown in Fig. 21, the mobilization of myeloid cells (non-lymphocytes) by GPC-100 is further enhanced by the addition of 7 days of propranolol pretreatment. CD11b+ myeloid cells and F4 / 80 macrophages are mobilized by GPC-100 and propranolol. Cell therapies targeting anti-tumor responses from myeloid cells are currently being investigated for oncology and other therapeutic fields. For example, see Fig. 22, which illustrates the mobilization of M1 and M2 macrophages, MDSCs, etc., in naive and tumor-bearing mice.

[0717] Example 9: GPC-100 improves cell migration in cancer-bearing mice.

[0718] As shown in Table 7, GPC-100 mobilization was enhanced in tumor-bearing mice. For example, GPC-100 increased circulating CXCR4-positive macrophages (F4 / 80) 2.7-fold in naive mice, but increased them 4.2-fold in tumor-bearing mice.

[0719] GPC-100 mobilization was also enhanced in tumor-bearing mice, which showed 7 times more macrophages in the circulation compared to naive mice.

[0720] As shown in Figures 22a-22b, GPC-100 also generated greater mobilization for CD11b myeloid cells, particularly in tumor-bearing mice (CT26-colon cancer).

[0721]

[0722] Example 10: A randomized, open-label, double-blind, placebo-controlled, multicenter, phase 2 clinical trial to evaluate combination therapy of GPC-100 and G-CSF in patients with multiple myeloma (MM), non-Hodgkin lymphoma (NHL), and / or Hodgkin's disease (HD).

[0723] This randomized, open-label, double-blind, placebo-controlled, multicenter, Phase 2 clinical trial (NCT02104427) evaluated the HSC mobilization and safety of GPC-100 and G-CSF combination therapy in patients with MM, NHL, or HD. Administration of GPC-100 resulted in the potent mobilization of CD34+ HSCs and CD133+ epithelial progenitor cells from the bone marrow to the PB. At peak levels, GPC-100 increased CD34+ cell numbers 3 to 14-fold compared to baseline levels (Sukhtankar 2023b). Phase 1 study data, as well as nonclinical safety, pharmacology, and toxicology results available to date, demonstrated sufficient safety and efficacy to support the further clinical development of GPC-100.

[0724] The primary objective of this Phase 2 study was to investigate HSC mobilization by GPC-100 (3.14 mg / kg) in combination with G-CSF in patients with MM, NHL, or HD. The total number of HSCs collected from leukocyte apheresis products was evaluated for each patient. Interestingly, faster mobilization kinetics were observed when GPC-100 was combined with G-CSF compared to that observed with GPC-100 alone. Additionally, increased lymphocyte mobilization by GPC-100 and G-CSF was observed.

[0725] Research Design MethodsThis clinical study was conducted in accordance with the ethical principles of the "Good Clinical Practice (GCP)" and the Declaration of Helsinki after obtaining signed informed consent from each patient. The protocol and its revisions, written subject information, informed consent forms, and other appropriate study-related materials were appropriately reviewed and approved.

[0726] The primary eligibility criteria included patients aged 18 to 75 years with confirmed MM, NHL, or HL, Eastern Medical Collaboration Group (ECOG) performance status 0-1, and potential candidates for HSC transplantation.

[0727] On the first day, eligible patients ≥5.0x10 6 To reach the target collection level of cells / kg, a subcutaneous (SC) injection of 10 μg / kg / day G-CSF was administered daily at 4:00 PM (±1 hour) as needed. Patients may have received G-CSF for up to 8 consecutive days. Starting on Day 5, patients received 3.14 mg / kg GPC-100 intravenously over 15 minutes, beginning at 8:30 AM (±1 hour), and ≥5.0x10 6 To reach the target collection level of cells / kg, leukocyte apheresis was performed daily as needed (typically 2 ± 1 hours after GPC-100 administration). Patients may have received GPC-100 for up to 5 consecutive days. Leukocyte apheresis may have continued for up to 5 sessions if necessary.

[0728] A continuous flow centrifuge device was used to collect peripheral stem cells during leukocyte apheresis. The volume of whole blood processed during each leukocyte apheresis procedure was 18 to 24 L. The leukocyte apheresis products were processed at the study site for CD34+ cell counting.

[0729] PB samples of circulating CD34+ cells were collected before each dose of GPC-100 (within 30 minutes of the start of administration), before leukocyte apheresis (within 30 minutes of the start of leukocyte apheresis), after leukocyte apheresis (within 30 minutes of the completion of leukocyte apheresis), and 4 hours (±20 minutes) and 6 hours (±20 minutes) after the completion of GPC-100 infusion.

[0730] The demographic and baseline characteristics of this study are summarized in Table 8. A total of 12 patients were enrolled, received G-CSF and GPC-100 3.14 mg / kg as planned, and completed the study. The majority of patients (75%) had MM. All enrolled patients had an ECOG status of grade 0 or 1. None of the patients had bone marrow involvement and had previously undergone stem cell collection.

[0731]

[0732] Example 11: Combination therapy with GPC-100 and G-CSF is safe and generally well tolerated in patients with multiple myeloma, non-Hodgkin lymphoma and / or Hodgkin's disease.

[0733] The safety evaluation of the combination therapy of GPC-100 and G-CSF was performed prior to administration and throughout the clinical study as described in Example 10. Specifically, the safety evaluation included the following: adverse events (AEs), including endogenous adverse events (TEAEs) and severe adverse events (SAEs); physical examination findings; clinical laboratory evaluations (chemistry, hematology, urinalysis); vital signs (blood pressure, heart rate, respiratory rate, body temperature); 12-lead electrocardiogram (ECG); pulse oximetry; and ECOG status. The sample size of this study was based on feasibility without formal power calculations. All safety and efficacy data were analyzed technically.

[0734] The primary efficacy endpoint is a total CD34+ cell count ≥5.0 x 10 within the first 4 leukocyte apheresis sessions. 6It was the frequency and proportion of patients in whom cells / kg were collected. For this endpoint, the CD34+ cell count for each patient was calculated as the sum of CD34+ cell counts collected from the (maximum) first four leukocyte apheresis sessions. A 95% accurate confidence interval (Ci) for the proportion of patients who met the target collection threshold was provided. The exact Ci, based on the binomial distribution, was constructed using the SAS PROC FREQ procedure.

[0735] ≥2.5 x10 6 CD34+ cells / kg and ≥5.0x10 6 The average number of leukocyte apheresis sessions required to collect CD34+ cells / kg was also evaluated.

[0736] As shown in Table 9, the administration of GPC-100 in combination with G-CSF was safe and generally well tolerated in patients with MM, NHL, or HD. A total of 7 patients reported 10 treatment-related adverse events (TEAEs). No severe adverse events (SAEs) were reported during this study. No patients discontinued treatment due to TEAEs. The number of patients with TEAEs was higher after the co-administration of GPC-100 and G-CSF (n=6) compared to G-CSF monotherapy (n=2) during the initial 4 days. The majority of TEAEs were grade 1 or 2, and there was a single grade 4 TEAE of thrombocytopenia, but this was assessed by the investigator as unrelated to the study drug. Two drug-related TEAEs were reported, including one patient with grade 2 bone pain after G-CSF monotherapy and one patient with grade 1 diarrhea after the co-administration of GPC-100 and G-CSF. There were no clinically significant changes in vital signs, ECG parameters, or physical examination during the study. Aside from the expected drug effect of increasing WBC counts, changes in other clinical laboratory parameters were generally unremarkable.

[0737]

[0738] Example 12: Combination therapy with GPC-100 and G-CSF in patients exhibits faster mobilization kinetics and increased lymphocyte mobilization compared to patients treated with GPC-100 alone.

[0739] The efficacy of combination therapy with GPC-100 and G-CSF was evaluated as described in Example 10. CD34 collected across all leukocyte apheresis sessions after co-administration of GPC-100 and G-CSF + The average total number of cells is 8.17 x 10⁶ 6 It was cells / kg (Table 10). Of the 12 enrolled patients, 5 underwent one leukocyte apheresis session to achieve the target cell count, and 7 underwent two sessions to achieve the target cell count. The average CD34 of MM patients + Number of cells (9.00 x 10⁻⁶ 6 cells / kg is the sum of NHL and HD patients (5.68 x 10⁻¹⁰ 6 It was higher than (cells / kg). Within one or two leukocyte apheresis sessions (average 1.4 sessions), 11 out of 12 patients (92%: 8 MM patients and 3 NHL or HD patients) had ≥ 5.0 x 10 6 CD34 cells / kg + The cell mobilization goal was achieved. All 8 MM patients ≥ 6.0x10 6 CD34 cells / kg + The target number of mobilized cells was achieved; among them, 6 patients achieved the target number after 1 session and 2 patients achieved the target after 2 sessions. Within a single leukocyte apheresis session, all patients except 1 (92%) had ≥2.5 x 10 6 CD34 cells / kg + The cell mobilization goal was achieved. One MM patient who did not meet the target cell count within one or two leukocyte apheresis sessions had CD34 up to one week prior to study entry. + I had been treated with lenalidomide (Cashen 2007), a drug known to lower cell yield.

[0740]

[0741] Circulating CD34 in the PB between Day 4 of G-CSF administration and GPC-100 administration + Changes in cell number were used to measure the pharmacodynamics of GPC-100. PB CD34 + The median number of cells increased 1.3-fold and 1.75-fold, 4 hours after GPC-100 infusion on Day 5 (Day 1 of leukocyte apheresis) and Day 6 (Day 2 of leukocyte apheresis), respectively, compared to before administration (the morning after G-CSF Day 4 and before GPC-100 administration) (Table 11). PB CD34 + Cell counts were relatively highest before leukocyte apheresis (1.9-fold and 2.1-fold on days 5 and 6, respectively), and returned to levels slightly higher than before administration after leukocyte apheresis. On day 6, PB CD34 at each sampling time point + The median number of cells was approximately 50% of that on Day 5. The number of CD34+ cells was not measured prior to the first administration of G-CSF (Figs. 23a-23b). CD34 in PB + Cellular changes can be graphed at each time point as the average for all patients (Fig. 23a) or the relative change for each individual patient (Fig. 23b). In both graphs, peak mobilization appears to occur approximately 1 hour after GPC-100 administration. As shown in Fig. 23b, CD34 for all 9 participating patients + The percentage of relative change in the peak is in the range of 45 minutes ± 15 minutes (or about 30 minutes to about 60 minutes).

[0742]

[0743] Example 13: Combination therapy with GPC-100, propranolol, and G-CSF is safe and well-tolerated in multiple myeloma (MM) patients receiving asct.

[0744] In this ongoing clinical trial, approximately 20 participants will be enrolled at 10 sites to evaluate the safety and clinical activity of GPC-100 according to a Bayesian Optimum Phase 2 (BOP2) design.

[0745] From Day 1 to Day 8, the patient will self-administer 30 mg of propranolol orally twice daily. From Day 3 to Day 7, the patient will also receive a subcutaneous injection of 10 mg / kg / day G-CSF daily in the afternoon. On Days 7 and 8, the patient will also receive a dose of 3.14 mg / kg of GPC-100 via IV and undergo leukocyte apheresis 45 minutes after drug administration. Participants may undergo optional additional days of treatment regimen and stem cell collection at the investigator's discretion to meet the institution's collection target standards. Participants will be monitored for adverse events (AEs) for up to 28 days after the last administration of GPC-100.

[0746] Preliminary results from four enrolled patients (age range: 60–71 years, median age: 67 years) demonstrate that combination therapy with GPC-100, propranolol, and G-CSF is safe and well-tolerated. Two of these patients had previously received four cycles of daratumumab-RVd. Of the other two patients, one received three cycles of Dara-R and the other received four cycles of RVd. A total of 45 all-grade AEs were observed, 71% of which occurred post-transplant. GPC-100-related AEs were grade 1 or lower and included flushing, chest tightness, abdominal pain, and nausea, consistent with previous observations. No diarrhea was reported during mobilization or leukocyte apheresis. No propranolol-related AEs were observed. 18% of all AEs were grade 3 or higher, all of which occurred post-transplant.

[0747] Example 14: Combination therapy with GPC-100, propranolol, and G-CSF in multiple myeloma patients undergoing ASCT despite the negative mobilization effects of daratumumab CD34 + Rapidly mobilizes cells.

[0748] This U.S.-only, open-label, multicenter Phase 2 clinical study (NCT05561751) evaluates the safety and transfer efficacy of MM patients undergoing ASCT treated with a combination of GPC-100, propranolol, and G-CSF. Methods and patient selection are described in Example 13. Transfer efficacy is defined as ≥2 × 10⁶ within two leukocyte apheresis sessions when the combination of GPC-100, propranolol, and G-CSF is administered. 6 CD34 + It is evaluated by measuring the proportion of patients who achieved cells / kg.

[0749] The preliminary results of combination therapy with GPC-100, propranolol, and G-CSF are promising. Table 12 and Figure 24 show that the primary objective of achieving minimum harvesting requirements was achieved for all MM patients. In particular, all of the four enrolled patients had ≥2×10 within two apheresis sessions. 6 CD34 + The primary endpoint of cells / kg (minimum harvest requirement) was achieved. 75% of these patients had ≥6×10 within two apheresis sessions. 6 CD34 + The primary endpoint of cells / kg (optimal harvest requirement) was achieved. Here, all four patients underwent ASCT. The timing of their ASCT performance is merely a logistical detail and is not affected by the efficacy of GPC-100. The median time to neutrophil engraftment was 12 days (range 11–15 days). Preliminary results showed that the combination therapy regimen CD34 for ASCT + It demonstrates that stem cells can be sufficiently mobilized.

[0750] Table 12 and Figure 24 also show CD34 in patients despite receiving a treatment (i.e., daratumumab treatment) where combination therapy may have a negative effect on mobilization. + It demonstrates that it is effective in mobilizing stem cells.

[0751] Table 12 and Figure 24 also show substantially improved mobilization dynamics compared to the FDA-approved AMD3100 or BL8040. While both AMD3100 and BL8040 require overnight pretreatment prior to leukocyte apheresis, the GPC-100 regimen described herein is significantly superior to existing methods because it allows for the administration of the mobilizing agent and leukocyte apheresis to be performed on the same day, and includes a step of administering the mobilizing agent approximately 30 to 60 minutes later than leukocyte apheresis (Figure 23b). Thus, combination therapy with GPC-100, propranolol, and G-CSF provides substantial improvements in the health of the bone marrow stem cell niche and offers a faster mobilization profile in patients such as MM patients seeking HSC transplantation.

[0752]

[0753] From the foregoing, it will be understood that while specific embodiments of the present invention have been described herein for illustrative purposes, various modifications may be made without departing from the scope of the invention. Accordingly, the present invention is not limited except as defined by the appended claims.

[0754] abbreviation

[0755] Unless otherwise specified, the following include abbreviations for terms disclosed herein: ACKR3 (Atypical Chemokine Receptor 3); ACT (Adoptive Cell Therapy); ADCYAP1R1 (ADCYAP Receptor Type I); ADCYAP1R1 (Adenylate Cyclase Activated Polypeptide 1 (Pituitary) Receptor Type I); AdHTS (Adenovirus High Efficiency System); ADORA2B (Adenosine Receptor A2b or Adenosine A2b Receptor); ADORA3 (Adenosine α3 Receptor); ADRA1A (Adrenergic Receptor Alpha 1a); ADRB2 (Adrenergic Receptor Beta 2); ADRB2, β2AR, or Gene ADRB2 (Beta-2 Adrenergic Receptor); AEs (Adverse Effects); ALL (Acute Lymphocytic Leukemia); AML (Acute Myeloid Leukemia); APLNR (Apelin Receptor); ARF (Alternative Read Frame); ASCT (Autologous Stem Cell Transplantation); ATCC (American Type Culture Collection); Beta-blockers (Beta-adrenergic inhibitors); BFU-E (Erythrocyte Burst-forming Unit); BiFC (Bimolecule Fluorescence Complementary); BM (Bone Marrow); BRET (Bioluminescent Resonance Energy Transfer); BSA (Bovine Serum Albumin); C5AR1 (Complement c5a Receptor 1); CALCR (Calcitonin Receptor); CAR (Chimeric Antigen Receptor); CBC (Complete Blood Count); CBC (Complete Blood Count); CCR2 (CC Chemokine Receptor Type 2 or Chemokine (CC Motif) Receptor 2); CCR5 (CC Chemokine Receptor Type 5 or Chemokine (CC Motif) Receptor 5); CFC (Continuous Flow Centrifugation); CFU (Colony-forming Unit); CFU-GEMM (Multivational Colony-forming Unit); CFU-GM (Granulocyte-Macrophage Colony-forming Unit); CFU-GM (Granulocyte-monocyte progenitor cell); CHRM1 (Cholinergic receptor muscarinic 1); CLL (Chronic lymphocytic leukemia); CMKLR1 (Chemerin chemokine-like receptor 1); CML (Chronic myeloid leukemia); COPD (Chronic obstructive pulmonary disease); CPP (Cell penetration peptide); CRE (Camp response factor);CSC (Cancer Stem Cell); CT (Computed Tomography); Ct (Threshold Cycle); CTLA-4 (Cytotoxic T-Lymphocyte-Associated Antigen 4); CTRA (Conserved Transcription Response to Adversity); CXCL12 (CXC Motif Chemokine Ligand 12, also known as SDF-1); CXCR4 (CXC Receptor 4); EDNRB (Endothelin Receptor Type b); ELISA (Enzyme-Linked Immunoassay); FACS (Fluorescence Activated Cell Sorting); FBS (Fetal Bovine Serum); FFPE (Formalin-Fixed Paraffin Embedded); FRET (Fluorescence Resonance Energy Transfer); GALR1 (Galanin Receptor 1); GBM (Glioblastoma); GCGR (Glucagon Receptor); GCID (Genecards Identification); G-CSF (Granulocyte Colony-Stimulating Factor); GFP (Green Fluorescent Protein); GPCR (G Protein-Coupling Receptor); GPCR-HIT (GPCR Heteromer Identification Technology); HCC (Hepatocellular Carcinoma); HD (Hodgkin's Disease); HGNC (Hugo Gene Nomenclature Committee); HIV (Human Immunodeficiency Virus); HL (Hodgkin Lymphoma); HPP-CFC (Highly Proliferative Colony-forming Cell); HRH1 (Histamine Receptor H1); HSC (Hematopoietic Progenitor Cell); HSCs (Hematopoietic Stem Cells); HTRF (Homogeneous Time-Resolved Fluorescence); IACUC (Institutional Animal Research Ethics Committee); Ki (Inhibition Constant); LT-HSC (Organ Reconstitution Ability); MAP (Model Amphiphilic Peptide); MDS (Myelodysplastic Syndrome); MLNR (Motilin Receptor); MM (Multiple Myeloma); MOI (Infection Multiplicity); MOR (μ-Opioid Receptor); NC-IUPHAR (International Union for Basic and Clinical Pharmacology Receptor Nomenclature and Drug Classification Committee); NFAT-RE (Nuclear Factor of Activated T-Cell Response Factor); NGS (Next-Generation Sequencing); NHL (Non-Hodgkin Lymphoma); NK (Natural Killer); NLS (Nuclear Localization Signaling); NSCLC (Non-Small Cell Lung Cancer); NTSR1 (Neurotensin Receptor 1); OPRD (δ-Opioid Receptor); PB (Peripheral Blood); PD-1 (Programmed Cell Death Protein 1); PDC (Patient-Derived Cell);PD-L1 (Programmed Apoptosis Ligand 1); PDX (Patient-Derived Xenograft); PET (Positron Emission Tomography); PET / CT (Positron Emission Tomography / Computed Tomography); PLA (Proximal Ligation Assay); PLA (Proximal Ligation Assay); Pro (Profranolol); PTGER2 (Prostaglandin E Receptor 2); PTGER3 (Prostaglandin E Receptor 3); RT-qPCR (Reverse Transcription-Quantitative PCR); RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction); SAE (Severe Adverse Event); SCD (Sickle Cell Disease); SCLC (Small Cell Lung Cancer); SCM (Enhanced Stem Cell Mobilization); SDF-1 (Stromal Cell-Derived Factor 1, also known as CXCL12); SLE (Systemic Lupus Erythematosus); SLL (Small Lymphocytic Lymphoma); SPECT (Single Photon Emission Computed Tomography); SPECT (Single Photon Emission Computed Tomography); SRE (Seroreactive Factor Response Element); SRF-RE (Seroreactive Factor Response Element); SSTR2 (Somatostatin Receptor 2); TACR3 (Tachykinin Receptor 3); TCR (T-Cell Receptor); TEAE (Treatment-Related Adverse Events); Th (T Helper Cell), Th1 (T Helper 1 Cell), Th (T Helper 2 Cell), TIL (Tumor-Infiltrating Lymphocyte); TME (Tumor Microenvironment); TR-FRET (Time-Resolved FRET); VC (C-terminal Fragment of Venus); VEGF (Vascular Endothelial Growth Factor); VN (N-terminal Fragment of Venus); VSMC (Vascular Smooth Muscle Cell); WBC (White Blood Cell); WHIM Syndrome (Warts, Hypogammaglobulinemia, Infection, and Myelostasis); and YFP (Yellow Fluorescent Protein).;

[0756] References

[0757]

[0758]

[0759]

[0760]

[0761]

[0762]

[0763]

[0764]

[0765]

[0766]

[0767]

[0768]

[0769]

[0770]

[0771]

[0772]

[0773]

[0774]

[0775]

[0776] Listed implementation examples

[0777] Implementation Example 1. A method for recruiting cells in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject.

[0778] Implementation Example 2. A method for inducing cell recruitment in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject.

[0779] Implementation Example 3. A method for improving apheresis by inducing cell mobilization in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in a subject.

[0780] Implementation Example 4. A method for improving apheresis by mobilizing cells from a subject, the method comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject.

[0781] Implementation Example 5. A method in any one of embodiments 1 to 4, wherein the cell is a stem cell.

[0782] Implementation Example 6. A method in any one of embodiments 1 to 5, wherein the cell is an LSK(Lin-Sca1+c-Kit+) cell.

[0783] Implementation Example 7. In Embodiment 6, the LSK cells are long-term reconstituted hematopoietic stem cells (LT-HSC), method.

[0784] Implementation Example 8. In Embodiment 7, the LT-HSC is a CD150+, CD34-, or CD48+ LSK cell.

[0785] Implementation Example 9. A method in any one of embodiments 1 to 8, wherein the step of blocking CXCR4 signaling includes the step of administering a CXCR4 inhibitor to a subject.

[0786] Implementation Example 10. In Embodiment 9, the CXCR4 inhibitor forms an ion interaction with Arg188 on CXCR4.

[0787] Example 11. In Embodiment 9 or 10, the CXCR4 inhibitor forms a hydrogen bond with Gln200 on CXCR4.

[0788] Implementation Example 12. A method in any one of embodiments 9 to 11, wherein the CXCR4 inhibitor has a higher binding affinity for CXCL12 than AMD3100.

[0789] Example 13. A method in any one of embodiments 9 to 12, wherein the CXCR4 inhibitor has an inhibition constant (Ki) lower than that of AMD3100.

[0790] Example 14. A method in any one of embodiments 9 to 13, wherein the CXCR4 inhibitor has an inhibition constant at least twice lower than that of AMD3100.

[0791] Example 15. A method in any one of embodiments 9 to 14, wherein the CXCR4 inhibitor inhibits CXCL12-induced calcium flux to a level equivalent to or higher than that of AMD3100.

[0792] Example 16. A method in any one of embodiments 9 to 15, wherein the CXCR4 inhibitor inhibits CXCL12-induced β-arrestin recruitment to a level equivalent to or higher than that of AMD3100.

[0793] Example 17. A method in any one of embodiments 9 to 16, wherein the CXCR4 inhibitor inhibits CXCL12-induced migration of cancer cells to a level equivalent to or higher than that of AMD3100.

[0794] Example 18. A method in any one of embodiments 9 to 17, wherein the CXCR4 inhibitor inhibits CXCL12-induced migration of U937 and MM.1S cells to a level equivalent to or higher than that of AMD3100.

[0795] Example 19. A method in any one of embodiments 9 to 18, wherein the CXCR4 inhibitor inhibits calcium flux induced by CXCL12 and epinephrine.

[0796] Example 20. A method in any one of embodiments 9 to 19, wherein blocking beta-adrenergic receptor signaling inhibits calcium flux and / or cell migration induced by CXCL12 and epinephrine.

[0797] Example 21. A method in any one of embodiments 9 to 19, wherein the blocking of propranolol inhibits calcium flux and / or cell migration induced by CXCL12 and epinephrine.

[0798] Example 22. A method in any one of embodiments 1 to 21, wherein the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in a subject induces an increased amount of long-term reconstituted hematopoietic stem cell (LT-HSC) mobilization compared to the amount of LT-HSC mobilization induced by a CXCR4 inhibitor and G-CSF alone.

[0799] Example 23. In any one of embodiments 1 to 22, the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in a subject is a method of recruiting LT-HSCs in an increased amount compared to the amount of LT-HSC recruitment induced by a CXCR4 inhibitor and G-CSF alone.

[0800] Example 24. In Embodiment 22 or 23, the LT-HSC expresses CD150.

[0801] Example 25. A method in which, in Embodiment 22 or 23, LT-HSC does not express CD34.

[0802] Example 26. In Embodiment 22 or 23, the LT-HSC expresses CD48.

[0803] Example 27. A method comprising, in any one of embodiments 1 to 26, an additional step of administering G-CSF.

[0804] Example 28. In Embodiment 27, a method wherein a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induce an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor and G-CSF alone.

[0805] Example 29. In Embodiment 27, a method wherein a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF mobilize cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor and G-CSF alone.

[0806] Example 30. A method in any one of embodiments 1 to 29, wherein the step of blocking beta-adrenergic receptor signaling is performed before the step of blocking CXCR4 signaling.

[0807] Example 31. A method in any one of embodiments 1 to 30, wherein the step of blocking beta-adrenergic receptor signaling continues even after the step of blocking CXCR4 signaling is terminated.

[0808] Implementation Example 32. A method in any one of embodiments 1 to 31, wherein the step of blocking CXCR4 signaling includes the step of administering a CXCR4 inhibitor to a subject.

[0809] Implementation Example 33. A method in any one of embodiments 1 to 32, wherein the step of blocking beta-adrenergic receptor signaling comprises the step of administering a beta-adrenergic receptor inhibitor to a subject.

[0810] Implementation Example 34. In Embodiment 33, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor.

[0811] Example 35. In Embodiment 33 or 34, the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenorolol, atenolol, betaxolol, bufranolol, butoxamine, carazolol, carvedilol, CGP 12177, cycloprorolol, ICI 118551, ICYP, labetalol, levobetaxolol, levovunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, and timolol.

[0812] Example 36. In Embodiment 35, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551.

[0813] Example 37. In Embodiment 36, the beta-adrenergic receptor inhibitor is propranolol, method.

[0814] Example 38. In any one of Embodiments 9 to 37, the CXCR4 inhibitor is ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549, D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GSTNT21MP, Isothiourea-1a, Isothiourea-1t (IT1t), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[11C]methyl-AMD3465, POL6326, SDF-1 l-9[P2G] dimer, SDFl P2G, T134, T140, T22, TC 14012, GPC-100 (burixafor, or TG-0054), USL311, viral macrophage inflammation protein-II (vMIP-11), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixater, [99mTc]O2-AMD3100, [177Lu]pentixater, ulocuplumab (MDX1338 / BMS-936564), AD-214, LY2624587, PF-06747143, A method selected from the group consisting of POL6326, MB1707, and 508MC1 (compound 26).

[0815] Example 39. In Embodiment 38, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, GPC-100 (Bulixafor, or TG-0054), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564).

[0816] Implementation Example 40. In Embodiment 39, the CXCR4 inhibitor is GPC-100 (burixaphor, or TG-0054), method.

[0817] Implementation Example 41. In Embodiment 39, the CXCR4 inhibitor is AMD3100 (Plerixafor, or Mozobil), method.

[0818] Implementation Example 42. In Embodiment 39, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564), method.

[0819] Implementation Example 43. A method in any one of embodiments 33 to 42, wherein the step of administering a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor to a subject comprises the step of administering GPC-100 (burixafor, or TG-0054) and propranolol.

[0820] Example 44. A method in any one of embodiments 33 to 42, wherein the step of administering a CXCR4 inhibitor beta-adrenergic receptor inhibitor to a subject comprises the step of administering AMD3100 (plerixafor, or mozobil) and propranolol.

[0821] Example 45. A method in any one of embodiments 33 to 42, wherein the step of administering a CXCR4 inhibitor beta-adrenergic receptor inhibitor to a subject comprises the step of administering ulocuplumab (MDX1338 / BMS-936564) and propranolol.

[0822] Example 46. A method in any one of embodiments 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone.

[0823] Example 47. In any one of embodiments 27 to 42, the step of administering the combination of a CXCR4 inhibitor and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone.

[0824] Implementation Example 48. In any one of embodiments 27 to 42, the step of administering a combination of a CXCR4 inhibitor and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor alone.

[0825] Implementation Example 49. A method in any one of embodiments 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone.

[0826] Example 50. In any one of embodiments 27 to 42, the step of administering a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone.

[0827] Example 51. In any one of embodiments 27 to 42, the step of administering a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor alone.

[0828] Example 52. A method in any one of embodiments 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor alone.

[0829] Implementation Example 53. In any one of embodiments 27 to 42, the step of administering a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF is a method of mobilizing cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor alone.

[0830] Example 54. In any one of embodiments 27 to 42, the step of administering a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor alone.

[0831] Example 55. In any one of embodiments 27 to 42, the step of administering a combination of GPC-100 (burixafor, or TG-0054) and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF, a method.

[0832] Example 56. In any one of embodiments 27 to 42, the step of administering a combination of GPC-100 (burixafor, or TG-0054) and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF.

[0833] Example 57. In any one of embodiments 27 to 42, the step of administering a combination of GPC-100 (burixafor, or TG-0054) and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by AMD3100 (plerixafor, or mozobil) and G-CSF.

[0834] Example 58. A method in any one of embodiments 47 to 57, wherein the increased amount of cell mobilization or component aggregation is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry analysis, and colony forming unit (CFU) analysis.

[0835] Example 59. In Embodiment 58, the increased amount of cell mobilization or component collection is measured by flow cytometry.

[0836] Example 60. In Embodiment 59, the method is performed by flow cytometry analysis on (Lin-Scal+c-Kit+) LSK cells.

[0837] Example 61. In Embodiment 58, the increased amount of cell mobilization or component collection is measured by colony-forming unit (CFU) analysis.

[0838] Example 62. A method in any one of embodiments 1 to 61, wherein the subject has a CXCR4 protomer in a cell.

[0839] Example 63. A method in any one of embodiments 1 to 62, wherein the subject has an ADRB2 protomer in a cell.

[0840] Example 64. A method in any one of embodiments 1 to 63, wherein the subject has a CXCR4 protomer and an ADRB2 protomer in a cell.

[0841] Example 65. In Embodiment 64, the method wherein the subject has a CXCR4-ADRB2 heteromer within the cell.

[0842] Example 66. In Embodiment 65, a method comprising: i) the CXCR4-ADRB2 heteromer having an increased amount of downstream calcium mobilization compared to downstream calcium mobilization from the CXCR4 protomer or ADRB2 protomer; and ii) the administered inhibitor combination inhibiting the enhanced downstream calcium mobilization from the CXCR4-ADRB2 heteromer within the stem cell.

[0843] Example 67. A method in any one of embodiments 5 to 66, wherein the stem cell is selected from the group consisting of hematopoietic stem cells, hematopoietic progenitor cells, mesenchymal stem cells, endothelial progenitor cells, neural stem cells, epithelial stem cells, skin stem cells, and cancer stem cells.

[0844] Example 68. In Embodiment 67, the stem cell is a hematopoietic stem cell or a hematopoietic progenitor cell.

[0845] Example 69. In Embodiment 68, a method in which hematopoietic stem cells or hematopoietic progenitor cells are mobilized from bone marrow to peripheral blood (PB).

[0846] Example 70. In Embodiment 69, the mobilized hematopoietic stem cells or hematopoietic progenitor cells are collected for transplantation into a patient with cancer.

[0847] Example 71. In Embodiment 70, the cancer is selected from the group consisting of lymphoma, leukemia, and myeloma.

[0848] Example 72. In Embodiment 71, the cancer is non-Hodgkin lymphoma (NHL), Hodgkin's disease (HD) or Hodgkin lymphoma (HL), acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), multiple myeloma (MM) or Burkitt lymphoma.

[0849] Example 73. A method in any one of embodiments 5 to 66, wherein the stem cell is a mesenchymal stem cell.

[0850] Example 74. In Embodiment 73, a method in which mesenchymal stem cells are mobilized from bone marrow to peripheral blood.

[0851] Example 75. In Embodiment 74, a method in which mesenchymal stem cells are mobilized for the treatment of a condition selected from the group consisting of nervous system disorders, cardiac ischemia, myocardial infarction, diabetes mellitus, tissue repair, bone and cartilage diseases, autoimmune diseases, graft-versus-host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

[0852] Example 76. A method in any one of embodiments 5 to 66, wherein the stem cell is a cancer stem cell.

[0853] Example 77. In Embodiment 76, a method in which cancer stem cells are mobilized into the blood.

[0854] Example 78. In Embodiment 76 or 77, a method in which cancer stem cells are mobilized for the treatment of cancer.

[0855] Example 79. A method in any one of embodiments 5 to 66, wherein the cell is an immune cell.

[0856] Example 80. In Embodiment 79, the immune cell is a white blood cell (WBC).

[0857] Example 81. In the embodiment 80, the WBC is a lymphocyte, method.

[0858] Example 82. In Embodiment 81, the lymphocytes are selected from the group consisting of T cells, B cells, and natural killer (NK) cells.

[0859] Example 83. In Embodiment 82, the lymphocyte is a T cell.

[0860] Example 84. In Embodiment 82, the lymphocytes are natural killer (NK) cells.

[0861] Example 85. In the embodiment 80, the WBC is a granulocyte, method.

[0862] Example 86. In Embodiment 85, the granulocytes are selected from the group consisting of neutrophils, eosinophils, and basophils.

[0863] Example 87. In Embodiment 86, the granulocytes are neutrophils.

[0864] Example 88. In the embodiment 81, the WBC is a monocyte, method.

[0865] Example 89. A method in any one of embodiments 79 to 88, wherein immune cells are mobilized from bone marrow to peripheral blood.

[0866] Example 90. A method in which, in any one of paragraphs 79 to 88, immune cells are mobilized from the spleen to peripheral blood.

[0867] Example 91. A method in which, in any one of paragraphs 79 to 88, immune cells are mobilized from the thymus to peripheral blood.

[0868] Example 92. A method in any one of embodiments 79 to 88, wherein immune cells are mobilized from lymph nodes to peripheral blood.

[0869] Example 93. A method in any one of embodiments 79 to 88, wherein the mobilized immune cells are used in adoptive cell therapy (ACT).

[0870] Example 94. In Embodiment 93, the adoptive cell therapy (ACT) is a chimeric antigen receptor (CAR) T cell therapy.

[0871] Example 95. In Embodiment 93, the adoptive cell therapy (ACT) is a natural killer (NK) cell therapy.

[0872] Example 96. In Embodiment 93, the adoptive cell therapy (ACT) is a method in which the adoptive cell therapy (ACT) is an engineered T-cell receptor (TCR) therapy.

[0873] Example 97. In Embodiment 93, the adoptive cell therapy (ACT) is a tumor-infiltrating lymphocyte (TIL) therapy.

[0874] Example 98. As a method of preparing a subject for treatment, the method comprises the following:

[0875] Step of blocking CXCR4 in the target,

[0876] A step of blocking beta-adrenergic receptors in a subject, or

[0877] Step of blocking CXCR4 inhibitors and beta-adrenergic receptors in a subject.

[0878] Example 99. In Example 98,

[0879] Blocking of CXCR4 is achieved through ionic interaction with Arg188 on CXCR4; and / or

[0880] Blocking of CXCR4 is achieved through hydrogen bonding with Gln200 on CXCR4; and / or any combination thereof, method.

[0881] Example 100. As a treatment method for a subject in need, the method comprises the following:

[0882] Step of blocking CXCR4 in the target,

[0883] A step of blocking beta-adrenergic receptors in a subject, or

[0884] Step of blocking CXCR4 inhibitors and beta-adrenergic receptors in a subject.

[0885] Example 101. In Example 100,

[0886] Blocking of CXCR4 is achieved through ionic interaction with Arg188 on CXCR4; and / or

[0887] Blocking of CXCR4 is achieved through hydrogen bonding with Gln200 on CXCR4; and / or any combination thereof, method.

[0888] Example 102. A method for preparing a subject for treatment, comprising the step of administering the following effective amount:

[0889] CXCR4 inhibitor,

[0890] Beta-adrenergic receptor inhibitors, or

[0891] Their combination.

[0892] Example 103. A method for qualifying a subject for treatment, comprising the step of administering the following effective amount:

[0893] CXCR4 inhibitor,

[0894] Beta-adrenergic receptor inhibitors, or

[0895] Their combination.

[0896] Example 104. A method for treating a subject in need, comprising the step of administering the following effective amount:

[0897] CXCR4 inhibitor,

[0898] Beta-adrenergic receptor inhibitors, or

[0899] Their combination.

[0900] Example 105. Use of GPC-100 for the manufacture of a drug for the treatment of cancer, or neurological disorders, or any combination thereof.

[0901] Example 106. In any one of the embodiments 98 to the immediately above, the treatment further comprises the step of administering the following effective amount:

[0902] Beta-adrenergic receptor inhibitors, or

[0903] G-CSF, or

[0904] Lenalidomide, or

[0905] Daratumumab, or

[0906] Velacade, or

[0907] Dexamethasone, or

[0908] Cyclophosphamide, or

[0909] Bortezomib, or

[0910] Their combination.

[0911] Example 107. A method in any one of the embodiments 98 to the immediately above, wherein the subject is pretreated by administering the following effective amount:

[0912] Beta-adrenergic receptor inhibitors, or

[0913] G-CSF, or

[0914] Lenalidomide, or

[0915] Daratumumab, or

[0916] Velacade, or

[0917] Dexamethasone, or

[0918] Cyclophosphamide, or

[0919] Bortezomib, or

[0920] Their combination.

[0921] Example 108. In any one of the embodiments 98 to the immediately above, the treatment comprises one or more of the following:

[0922] apheresis step; or

[0923] Leukocyte apheresis step; or

[0924] Peripheral blood collection step; or

[0925] As a cell transplantation step,

[0926] The cells are selectively stem cells, and

[0927] The cells are selectively hematopoietic progenitor cells (HSCs), and

[0928] The cell is selectively a T cell, or a combination thereof, step; and / or

[0929] CAR-T therapy stage; or

[0930] Their combination.

[0931] Example 109. A method comprising, in any one of the embodiments 98 to the immediately above, the step of administering an effective amount of G-CSF.

[0932] Implementation Example 110. In any one of the embodiments 98 to the immediately above,

[0933] The subject exhibits or is diagnosed with symptoms of cancer, or neurological disorders, or any combination thereof; and / or

[0934] Cancer is selected from the group consisting of lymphoma, leukemia, and multiple myeloma; and / or

[0935] The subject exhibits symptoms of blood cancer or is diagnosed with it; and / or

[0936] The cancer is selected from the group consisting of non-Hodgkin lymphoma (NHL), Hodgkin's disease (HD) or Hodgkin lymphoma (HL), acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), multiple myeloma (MM), and Burkitt lymphoma; and / or

[0937] The cancer is HD; and / or

[0938] The arm is NHL and; and / or

[0939] The cancer is AML; and / or

[0940] Cancer is ALL and; and / or

[0941] The cancer is MM; and / or

[0942] Neurological disorders are selected from the group consisting of neurological disorders, cardiac ischemia, myocardial infarction, diabetes mellitus, tissue repair, bone and cartilage diseases, autoimmune diseases, graft-versus-host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis; and / or

[0943] The subject is a human, method.

[0944] Example 111. In any one of the embodiments 98 to the immediately above,

[0945] The subject requires autologous stem cell transplantation (ASCT);

[0946] The treatment is a method called autologous stem cell transplantation (ASCT).

[0947] Example 112. In any one of the embodiments 98 to the immediately above,

[0948] Here:

[0949] CXCR4 inhibitors form ion interactions with Arg188 on CXCR4, or

[0950] CXCR4 inhibitors form hydrogen bonds with Gln200 on CXCR4, or

[0951] Any combination of these; and / or

[0952] Here:

[0953] CXCR4 inhibitors have a higher binding affinity for CXCL12 than AMD3100, or

[0954] CXCR4 inhibitors have a lower inhibition constant (Ki) than AMD3100, or

[0955] A CXCR4 inhibitor having an inhibition constant at least twice lower than AMD3100, method.

[0956] Example 113. In any one of the embodiments 98 to the immediately above,

[0957] CXCR4 inhibitors are:

[0958] ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (Plericsaphor, or Mozobil), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549, D-[Lys3]GHRP-6, FC122, FC131, GMI-1359, GSK812397, GSTNT21MP, Isothiourea-1a, Isothiourea-1t (IT1t), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[11C]methyl-AMD3465, POL6326, SDF-1 l-9[P2G] dimer, SDFl P2G, T134, T140, T22, TC 14012, GPC-100 (burixafor, or TG-0054), USL311, viral macrophage inflammation protein-II (vMIP-11), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixater, [99mTc]O2-AMD3100, [177Lu]pentixater, ulocuplumab (MDX1338 / BMS-936564), AD-214, LY2624587, PF-06747143, POL6326. Selected from the group consisting of MB1707, 508MC1 (Compound 26), and derivatives and analogs thereof; and / or

[0959] Selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, GPC-100 (Bulixafor, or TG-0054), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564), and derivatives and analogs thereof; and / or

[0960] ulocuflumab (MDX1338 / BMS-936564) and / or

[0961] AMD3100 (Pleric Sapor, or Mojobil) and / or

[0962] BL8040 and; and / or

[0963] GPC-100 (Buriksapor, or TG-0054) and; and / or

[0964] Beta-adrenergic receptor inhibitors are:

[0965] It is an ADRB2 inhibitor, and / or

[0966] Selected from the group consisting of alfrenollol, atenolol, betaxolol, bufranolol, butoxamine, carazolol, carvedilol, CGP 12177, cycloprollol, ICI 118551, ICYP, labetalol, levobetaxolol, levovunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, timolol, and derivatives and analogs thereof, and / or

[0967] Selected from the group consisting of propranolol, nadolol, and ICI 118551, and / or

[0968] Propranolol, method.

[0969] Example 114. In Example 113,

[0970] The CXCR4 inhibitor is GPC-100, or AMD3100, or BL8040, preferably GPC-100; and / or

[0971] The beta-adrenergic receptor inhibitor is propranolol, method.

[0972] Example 115. In any one of the embodiments 98 to the immediately above,

[0973] Here:

[0974] The effective dose of the CXCR inhibitor is:

[0975] It is a pharmacologically effective dose with clinically acceptable serious side effects, or

[0976] It is about 30 mg / kg to about 0.01 mg / kg, or

[0977] It is about 30 mg / kg to about 0.07 mg / kg, or

[0978] It is about 25 mg / kg to about 0.07 mg / kg, or

[0979] It is about 20 mg / kg to about 0.07 mg / kg, or

[0980] It is about 15 mg / kg to about 0.07 mg / kg, or

[0981] It is about 10 mg / kg to about 0.07 mg / kg, or

[0982] About 9 mg / kg to about 0.07 mg / kg, or

[0983] It is about 8 mg / kg to about 0.07 mg / kg, or

[0984] It is about 7 mg / kg to about 0.07 mg / kg, or

[0985] It is about 6 mg / kg to about 0.07 mg / kg, or

[0986] It is about 5 mg / kg to about 0.07 mg / kg, or

[0987] About 4.40 mg / kg to about 0.07 mg / kg, or

[0988] About 3.14 mg / kg to about 0.07 mg / kg, or

[0989] About 2.24 mg / kg to about 0.07 mg / kg, or

[0990] About 1.12 mg / kg to about 0.07 mg / kg, or

[0991] About 0.56 mg / kg to about 0.07 mg / kg, or

[0992] About 0.28 mg / kg to about 0.07 mg / kg, or

[0993] About 0.14 mg / kg to about 0.07 mg / kg, or

[0994] About 0.10 mg / kg to about 0.07 mg / kg, or

[0995] It is about 30 mg / kg to about 0.10 mg / kg, or

[0996] It is about 25 mg / kg to about 0.10 mg / kg, or

[0997] It is about 20 mg / kg to about 0.10 mg / kg, or

[0998] It is about 15 mg / kg to about 0.10 mg / kg, or

[0999] It is about 10 mg / kg to about 0.10 mg / kg, or

[1000] It is about 9 mg / kg to about 0.10 mg / kg, or

[1001] It is about 8 mg / kg to about 0.10 mg / kg, or

[1002] It is about 7 mg / kg to about 0.10 mg / kg, or

[1003] It is about 6 mg / kg to about 0.10 mg / kg, or

[1004] It is about 5 mg / kg to about 0.10 mg / kg, or

[1005] It is about 4.40 mg / kg to about 0.10 mg / kg, or

[1006] About 3.14 mg / kg to about 0.10 mg / kg, or

[1007] About 2.24 mg / kg to about 0.10 mg / kg, or

[1008] About 1.12 mg / kg to about 0.10 mg / kg, or

[1009] About 0.56 mg / kg to about 0.10 mg / kg, or

[1010] About 0.28 mg / kg to about 0.10 mg / kg, or

[1011] About 0.14 mg / kg to about 0.10 mg / kg, or

[1012] Approximately 4.40 mg / kg, or

[1013] Approximately 3.14 mg / kg, or

[1014] Approximately 2.24 mg / kg, or

[1015] Approximately 1.12 mg / kg, or

[1016] Approximately 0.56 mg / kg, or

[1017] Approximately 0.28 mg / kg, or

[1018] Approximately 0.14 mg / kg, or

[1019] Approximately 0.10 mg / kg, or

[1020] Approximately 0.07 mg / kg, or

[1021] CXCR4 inhibitors are:

[1022] Administered via a pharmacologically acceptable route, or

[1023] Administered orally, or

[1024] Administered subcutaneously, or

[1025] Administered intravenously; and / or

[1026] The duration of CXCR4 inhibitor administration is:

[1027] It is about 1 minute to about 30 minutes, or

[1028] It is about 5 minutes to about 20 minutes, or

[1029] It is about 10 minutes to about 18 minutes, or

[1030] It is about 15 minutes and; and / or

[1031] The time of administration for CXCR4 inhibitors is:

[1032] It is morning, or

[1033] It is about 7:00 AM to about 7 PM, or

[1034] It is around 7:30 AM to around 12:30 PM, or

[1035] It is around 7:30 AM to around 9:30 PM, or

[1036] It is about 8:30 AM and; and / or

[1037] The last dose of CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is:

[1038] When apheresis and / or leukocyte apheresis treatment begins

[1039] Administered about 120 hours to about 30 minutes prior, or

[1040] Administered about 72 hours to about 30 minutes prior, or

[1041] Administered about 24 hours to about 30 minutes prior, or

[1042] Administered about 14 hours to about 30 minutes prior, or

[1043] Administered about 13 hours to about 30 minutes prior, or

[1044] Administered about 13 hours to about 30 minutes prior, or

[1045] Administered about 12 hours to about 30 minutes prior, or

[1046] Administered about 11 hours to about 30 minutes prior, or

[1047] Administered about 10 hours to about 30 minutes prior, or

[1048] Administered about 9 hours to about 30 minutes prior, or

[1049] Administered about 8 hours to about 30 minutes prior, or

[1050] Administered about 7 hours to about 30 minutes prior, or

[1051] Administered about 6 hours to about 30 minutes prior, or

[1052] Administered about 5 hours to about 30 minutes prior, or

[1053] Administered about 4 hours to about 30 minutes prior, or

[1054] Administered about 3 hours to about 30 minutes prior, or

[1055] Administered about 2 hours to about 30 minutes prior, or

[1056] Administered about 90 minutes to about 30 minutes prior, or

[1057] Administered about 60 minutes to about 30 minutes prior, or

[1058] Administered about 45 minutes to about 30 minutes prior, or

[1059] Administered about 11 hours prior, or

[1060] Administered about 10 hours prior, or

[1061] Administered about 9 hours prior, or

[1062] Administered about 8 hours prior, or

[1063] Administered about 7 hours prior, or

[1064] Administered about 6 hours prior, or

[1065] Administered about 5 hours prior, or

[1066] Administered about 4 hours prior, or

[1067] Administered about 3 hours prior, or

[1068] Administered about 2 hours prior, or

[1069] Administered about 90 minutes prior, or

[1070] Administered about 60 minutes prior, or

[1071] Administered about 45 minutes prior, or

[1072] Administered about 30 minutes prior; and / or

[1073] CXCR4 inhibitors are administered once, twice, or three, four, or more than four times; and / or

[1074] CXCR4 inhibitors are administered one or fewer times, two or fewer times, three or fewer times, four or fewer times, or five or fewer times; and / or

[1075] CXCR4 inhibitors are administered for 1, 2, or 3, or 4, or for more than 4 days; and / or

[1076] CXCR4 inhibitors are administered for 1 day or less, 2 days or less, 3 days or less, 4 days or less, or 5 days or less; and / or

[1077] The apheresis and / or leukocyte apheresis treatment comprises about 1 to about 5 sessions, preferably about 1 to 2 sessions, more preferably 1 session; and / or

[1078] CXCR4 inhibitors are administered once daily; and / or

[1079] CXCR4 inhibitors are:

[1080] It is administered about 7 to about 11 days after the administration of propranolol, or

[1081] It is administered about 9 to 11 days after the administration of propranolol, or

[1082] Administered about 7 to 8 days after administration of propranolol; or

[1083] Administered on the 7th day after administration of propranolol, or

[1084] Administered on the 8th day after administration of propranolol; and / or

[1085] The first dose of the CXCR4 inhibitor is:

[1086] It is administered about 7 to about 11 days after the administration of propranolol, or

[1087] It is administered about 7 to 8 days after the administration of propranolol, or

[1088] Administered on the 7th day after administration of propranolol, or

[1089] Administered on the 8th day after administration of propranolol; and / or

[1090] The second dose of the CXCR4 inhibitor is:

[1091] It is administered about 7 to about 11 days after the administration of propranolol, or

[1092] It is administered about 9 to 11 days after the administration of propranolol, or

[1093] Administered on the 9th day after administration of propranolol, or

[1094] Administered on the 10th day after administration of propranolol, or

[1095] Administered on the 11th day after administration of propranolol; and / or

[1096] About 0.1 mg / kg to about 4.4 mg / kg of a CXCR4 inhibitor is administered intravenously once daily for 1 day or up to 5 days, preferably the last dose of the CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 30 to about 60 minutes before the start of apheresis and / or leukocyte apheresis treatment; and / or

[1097] Here:

[1098] The effective amount of GPC-100 is:

[1099] It is about 30 mg / kg to about 0.07 mg / kg, or

[1100] It is about 25 mg / kg to about 0.07 mg / kg, or

[1101] It is about 20 mg / kg to about 0.07 mg / kg, or

[1102] It is about 15 mg / kg to about 0.07 mg / kg, or

[1103] It is about 10 mg / kg to about 0.07 mg / kg, or

[1104] About 9 mg / kg to about 0.07 mg / kg, or

[1105] It is about 8 mg / kg to about 0.07 mg / kg, or

[1106] It is about 7 mg / kg to about 0.07 mg / kg, or

[1107] It is about 6 mg / kg to about 0.07 mg / kg, or

[1108] It is about 5 mg / kg to about 0.07 mg / kg, or

[1109] About 4.40 mg / kg to about 0.07 mg / kg, or

[1110] About 3.14 mg / kg to about 0.07 mg / kg, or

[1111] About 2.24 mg / kg to about 0.07 mg / kg, or

[1112] About 1.12 mg / kg to about 0.07 mg / kg, or

[1113] About 0.56 mg / kg to about 0.07 mg / kg, or

[1114] About 0.28 mg / kg to about 0.07 mg / kg, or

[1115] About 0.14 mg / kg to about 0.07 mg / kg, or

[1116] About 0.10 mg / kg to about 0.07 mg / kg, or

[1117] It is about 30 mg / kg to about 0.10 mg / kg, or

[1118] It is about 25 mg / kg to about 0.10 mg / kg, or

[1119] It is about 20 mg / kg to about 0.10 mg / kg, or

[1120] It is about 15 mg / kg to about 0.10 mg / kg, or

[1121] It is about 10 mg / kg to about 0.10 mg / kg, or

[1122] It is about 9 mg / kg to about 0.10 mg / kg, or

[1123] It is about 8 mg / kg to about 0.10 mg / kg, or

[1124] It is about 7 mg / kg to about 0.10 mg / kg, or

[1125] It is about 6 mg / kg to about 0.10 mg / kg, or

[1126] It is about 5 mg / kg to about 0.10 mg / kg, or

[1127] It is about 4.40 mg / kg to about 0.10 mg / kg, or

[1128] About 3.14 mg / kg to about 0.10 mg / kg, or

[1129] About 2.24 mg / kg to about 0.10 mg / kg, or

[1130] About 1.12 mg / kg to about 0.10 mg / kg, or

[1131] About 0.56 mg / kg to about 0.10 mg / kg, or

[1132] About 0.28 mg / kg to about 0.10 mg / kg, or

[1133] About 0.14 mg / kg to about 0.10 mg / kg, or

[1134] Approximately 4.40 mg / kg, or

[1135] Approximately 3.14 mg / kg, or

[1136] Approximately 2.24 mg / kg, or

[1137] Approximately 1.12 mg / kg, or

[1138] Approximately 0.56 mg / kg, or

[1139] Approximately 0.28 mg / kg, or

[1140] Approximately 0.14 mg / kg, or

[1141] Approximately 0.10 mg / kg, or

[1142] Approximately 0.07 mg / kg, or

[1143] GPC-100 is:

[1144] Administered via a pharmacologically acceptable route, or

[1145] Administered orally, or

[1146] Administered subcutaneously, or

[1147] Administered intravenously; and / or

[1148] The duration of GPC-100 administration is:

[1149] It is about 1 minute to about 30 minutes, or

[1150] It is about 5 minutes to about 20 minutes, or

[1151] It is about 10 minutes to about 18 minutes, or

[1152] It is about 15 minutes and; and / or

[1153] The administration time of GPC-100 is:

[1154] It is morning, or

[1155] It is about 7:00 AM to about 7 PM, or

[1156] It is around 7:30 AM to around 12:30 PM, or

[1157] It is around 7:30 AM to around 9:30 PM, or

[1158] It is about 8:30 AM and; and / or The last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is:

[1159] When apheresis and / or leukocyte apheresis treatment begins

[1160] Administered about 120 hours to about 30 minutes prior, or

[1161] Administered about 72 hours to about 30 minutes prior, or

[1162] Administered about 24 hours to about 30 minutes prior, or

[1163] Administered about 14 hours to about 30 minutes prior, or

[1164] Administered about 13 hours to about 30 minutes prior, or

[1165] Administered about 13 hours to about 30 minutes prior, or

[1166] Administered about 12 hours to about 30 minutes prior, or

[1167] Administered about 11 hours to about 30 minutes prior, or

[1168] Administered about 10 hours to about 30 minutes prior, or

[1169] Administered about 9 hours to about 30 minutes prior, or

[1170] Administered about 8 hours to about 30 minutes prior, or

[1171] Administered about 7 hours to about 30 minutes prior, or

[1172] Administered about 6 hours to about 30 minutes prior, or

[1173] Administered about 5 hours to about 30 minutes prior, or

[1174] Administered about 4 hours to about 30 minutes prior, or

[1175] Administered about 3 hours to about 30 minutes prior, or

[1176] Administered about 2 hours to about 30 minutes prior, or

[1177] Administered about 90 minutes to about 30 minutes prior, or

[1178] Administered about 60 minutes to about 30 minutes prior, or

[1179] Administered about 45 minutes to about 30 minutes prior, or

[1180] Administered about 11 hours prior, or

[1181] Administered about 10 hours prior, or

[1182] Administered about 9 hours prior, or

[1183] Administered about 8 hours prior, or

[1184] Administered about 7 hours prior, or

[1185] Administered about 6 hours prior, or

[1186] Administered about 5 hours prior, or

[1187] Administered about 4 hours prior, or

[1188] Administered about 3 hours prior, or

[1189] Administered about 2 hours prior, or

[1190] Administered about 90 minutes prior, or

[1191] Administered about 60 minutes prior, or

[1192] Administered about 45 minutes prior, or

[1193] Administered about 30 minutes prior; and / or

[1194] GPC-100 is administered 1, 2, or 3, 4, or more than 4 times; and / or

[1195] GPC-100 is administered 1 or fewer times, 2 or fewer times, 3 or fewer times, 4 or fewer times, or 5 or fewer times; and / or

[1196] GPC-100 is administered for 1, 2, or 3, or 4, or for more than 4 days; and / or

[1197] GPC-100 is administered for 1 day or less, 2 days or less, 3 days or less, 4 days or less, or 5 days or less; and / or

[1198] The apheresis and / or leukocyte apheresis treatment comprises about 1 to about 5 sessions, preferably about 1 to 2 sessions, more preferably 1 session; and / or

[1199] GPC-100 is administered once daily; and / or

[1200] GPC-100 is:

[1201] It is administered about 7 to about 11 days after the administration of propranolol, or

[1202] It is administered about 9 to 11 days after the administration of propranolol, or

[1203] Administered about 7 to 8 days after administration of propranolol; or

[1204] Administered on the 7th day after administration of propranolol, or

[1205] Administered on the 8th day after administration of propranolol; and / or

[1206] The first dose of GPC-100 is:

[1207] It is administered about 7 to about 11 days after the administration of propranolol, or

[1208] It is administered about 7 to 8 days after the administration of propranolol, or

[1209] Administered on the 7th day after administration of propranolol, or

[1210] Administered on the 8th day after administration of propranolol; and / or

[1211] The second dose of GPC-100 is:

[1212] It is administered about 7 to about 11 days after the administration of propranolol, or

[1213] It is administered about 9 to 11 days after the administration of propranolol, or

[1214] Administered on the 9th day after administration of propranolol, or

[1215] Administered on the 10th day after administration of propranolol, or

[1216] Administered on the 11th day after administration of propranolol; and / or

[1217] GPC-100 at a dose of about 0.1 mg / kg to about 4.4 mg / kg is administered intravenously once daily for 1 day or up to 5 days, preferably the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered about 30 to about 60 minutes before the start of apheresis and / or leukocyte apheresis treatment; and / or Here:

[1218] The effective capacity of the AMD3100 is:

[1219] It is about 10 μg / kg to about 480 μg / kg, or

[1220] About 40 μg / kg to about 480 μg / kg, or

[1221] About 80 μg / kg to about 480 μg / kg, or

[1222] It is about 40 μg / kg to about 480 μg / kg and is administered subcutaneously; or

[1223] It is about 80 μg / kg to about 480 μg / kg and is administered subcutaneously; or

[1224] It is approximately 40 μg / kg and is administered subcutaneously; or

[1225] It is approximately 80 μg / kg and is administered subcutaneously; or

[1226] It is approximately 480 μg / kg and is administered subcutaneously; or

[1227] It is about 10 μg / kg to about 80 μg / kg and is administered intravenously; or

[1228] It is about 20 μg / kg to about 80 μg / kg and is administered intravenously; or

[1229] It is about 40 μg / kg to about 80 μg / kg and is administered intravenously; or

[1230] It is approximately 10 μg / kg and is administered intravenously; or

[1231] It is approximately 20 μg / kg and is administered intravenously; or

[1232] It is approximately 40 μg / kg and is administered intravenously; or

[1233] It is approximately 80 μg / kg and is administered intravenously; and / or

[1234] AMD3100 is:

[1235] Administered via a pharmacologically acceptable route, or

[1236] Administered subcutaneously, or

[1237] Administered intravenously; and / or

[1238] The last dose of the AMD3100 inhibitor prior to the start of apheresis and / or leukocyte apheresis is:

[1239] When apheresis and / or leukocyte apheresis treatment begins

[1240] Administered about 120 hours to about 10 hours prior, or

[1241] Administered about 72 hours to about 10 hours prior, or

[1242] Administered about 24 hours to about 10 hours prior, or

[1243] Administered about 14 to 10 hours prior, or

[1244] Administered about 13 to 10 hours prior, or

[1245] Administered about 13 to 10 hours prior, or

[1246] Administered about 12 to 10 hours prior, or

[1247] Administered about 11 to 10 hours prior, or

[1248] Administered about 12 hours prior, or

[1249] Administered about 11 hours prior, or

[1250] Administered approximately 10 hours prior; and / or

[1251] AMD3100 is administered once daily; and / or

[1252] AMD3100 is administered 1, 2, 3, 4, or more than 4 times; and / or

[1253] AMD3100 is administered 5 times or less, 4 times or less, or 3 times or less, 2 times or less, or 1 time or less; and / or

[1254] AMD3100 is administered intravenously or subcutaneously, preferably subcutaneously, at a dose of about 10 μg / kg to about 480 μg / kg mg / kg once daily for 1 day or up to 5 days, preferably the last dose of AMD3100 prior to apheresis and / or leukocyte apheresis is administered at least about 10 hours prior to the initiation of apheresis and / or leukocyte apheresis treatment, or preferably about 12 to about 10 hours prior to the initiation of apheresis and / or leukocyte apheresis treatment; and / or

[1255] Here:

[1256] The effective amount of BL8040 is:

[1257] About 0.5 mg / kg to about 2.0 mg / kg, or

[1258] About 0.75 mg / kg to about 2.0 mg / kg, or

[1259] It is about 1.0 mg / kg to about 2.0 mg / kg, or

[1260] About 1.25 mg / kg to about 2.0 mg / kg, or

[1261] About 1.5 mg / kg to about 2.0 mg / kg, or

[1262] or about 0.5 mg / kg; or

[1263] Approximately 0.75 mg / kg or; or

[1264] or about 1 mg / kg; or

[1265] It is about 1.5 mg / kg and; and / or

[1266] BL8040 is:

[1267] Administered via a pharmacologically acceptable route, or

[1268] Administered subcutaneously, or

[1269] Administered intravenously; and / or

[1270] The last dose of BL8040 prior to the start of apheresis and / or leukocyte apheresis is:

[1271] When apheresis and / or leukocyte apheresis treatment begins

[1272] Administered about 120 hours to about 10 hours prior, or

[1273] Administered about 72 hours to about 10 hours prior, or

[1274] Administered about 24 hours to about 10 hours prior, or

[1275] Administered about 14 to 10 hours prior, or

[1276] Administered about 13 to 10 hours prior, or

[1277] Administered about 13 to 10 hours prior, or

[1278] Administered about 12 to 10 hours prior, or

[1279] Administered about 11 to 10 hours prior, or

[1280] Administered about 12 hours prior, or

[1281] Administered about 11 hours prior, or

[1282] Administered approximately 10 hours prior; and / or

[1283] BL8040 is administered once daily; and / or

[1284] BL8040 is administered 1, 2, 3, 4, or more than 4 times; and / or

[1285] BL8040 is administered 5 times or less, 4 times or less, or 3 times or less, 2 times or less, or 1 time or less; and / or

[1286] BL8040 is administered subcutaneously at about 1.5 mg / kg once daily for 1 day or up to 5 days, preferably the last dose of BL8040 prior to apheresis and / or leukocyte apheresis is administered at least about 10 hours before the apheresis and / or leukocyte apheresis treatment begins, preferably about 12 to about 10 hours before the apheresis and / or leukocyte apheresis treatment begins; and / or

[1287] Here:

[1288] The effective dose of a beta-adrenergic receptor inhibitor is:

[1289] It is a pharmacologically effective dose with clinically acceptable serious side effects, or

[1290] About 10 mg to about 480 mg per day, or

[1291] About 20 mg to about 480 mg per day, or

[1292] About 30 mg to about 480 mg per day, or

[1293] About 40 mg to about 480 mg per day, or

[1294] About 50 mg to about 480 mg per day, or

[1295] About 60 mg to about 480 mg per day, or

[1296] About 70 mg to about 480 mg per day, or

[1297] About 80 mg to about 480 mg per day, or

[1298] About 120 mg to about 480 mg per day, or

[1299] About 160 mg to about 480 mg per day, or

[1300] About 200 mg to about 480 mg per day, or

[1301] About 240 mg to about 480 mg per day, or

[1302] About 280 mg to about 480 mg per day, or

[1303] About 320 mg to about 480 mg per day, or

[1304] About 360 mg to about 480 mg per day, or

[1305] About 400 mg to about 480 mg per day, or

[1306] About 440 mg to about 480 mg per day, or

[1307] About 10 mg to about 480 mg per day, or

[1308] About 10 mg to about 450 mg per day, or

[1309] It is about 10 mg to about 420 mg per day, or

[1310] About 10 mg to about 390 mg per day, or

[1311] About 10 mg to about 360 mg per day, or

[1312] About 10 mg to about 330 mg per day, or

[1313] About 10 mg to about 300 mg per day, or

[1314] About 10 mg to about 270 mg per day, or

[1315] About 10 mg to about 240 mg per day, or

[1316] About 10 mg to about 210 mg per day, or

[1317] It is about 10 mg to about 180 mg per day, or

[1318] About 10 mg to about 150 mg per day, or

[1319] It is about 10 mg to about 120 mg per day, or

[1320] It is about 10 mg to about 90 mg per day, or

[1321] It is about 10 mg to about 60 mg per day, or

[1322] It is about 10 mg to about 30 mg per day, or

[1323] It is about 10 mg to about 90 mg per day, or

[1324] It is about 10 mg to about 60 mg per day, or

[1325] It is about 10 mg to about 30 mg per day, or

[1326] About 20 mg to about 480 mg per day, or

[1327] About 20 mg to about 450 mg per day, or

[1328] About 20 mg to about 440 mg per day, or

[1329] About 20 mg to about 410 mg per day, or

[1330] About 20 mg to about 400 mg per day, or

[1331] About 20 mg to about 370 mg per day, or

[1332] About 20 mg to about 360 mg per day, or

[1333] About 20 mg to about 330 mg per day, or

[1334] About 20 mg to about 320 mg per day, or

[1335] About 20 mg to about 290 mg per day, or

[1336] About 20 mg to about 280 mg per day, or

[1337] About 20 mg to about 250 mg per day, or

[1338] About 20 mg to about 240 mg per day, or

[1339] About 20 mg to about 210 mg per day, or

[1340] About 20 mg to about 200 mg per day, or

[1341] About 20 mg to about 170 mg per day, or

[1342] About 20 mg to about 160 mg per day, or

[1343] About 20 mg to about 130 mg per day, or

[1344] About 20 mg to about 120 mg per day, or

[1345] It is about 20 mg to about 90 mg per day, or

[1346] It is about 20 mg to about 80 mg per day, or

[1347] It is about 20 mg to about 50 mg per day, or

[1348] It is about 20 mg to about 40 mg per day, or

[1349] About 20 mg to about 160 mg per day, or

[1350] About 20 mg to about 130 mg per day, or

[1351] About 20 mg to about 120 mg per day, or

[1352] About 20 mg to about 100 mg per day, or

[1353] It is about 20 mg to about 80 mg per day, or

[1354] It is about 20 mg to about 70 mg per day, or

[1355] It is about 20 mg to about 40 mg per day, or

[1356] About 20 mg per day, or

[1357] About 40 mg per day, or

[1358] Approximately 60 mg per day, or

[1359] Approximately 80 mg per day, and / or

[1360] Beta-adrenergic receptor inhibitors are:

[1361] Administered via a pharmacologically acceptable route, or

[1362] Administered orally; and / or

[1363] Beta-adrenergic receptor inhibitors are administered once daily, or twice daily, or three times daily, or four times daily; and / or

[1364] Beta-adrenergic receptor inhibitors are:

[1365] Administered at intervals of approximately 7 to 12 hours, or

[1366] Administered at intervals of approximately 12 hours, or

[1367] Administered at intervals of approximately 11 hours, or

[1368] Administered at intervals of approximately 10 hours, or

[1369] Administered at intervals of approximately 9 hours, or

[1370] Administered at intervals of approximately 8 hours, or

[1371] Administered at intervals of approximately 7 hours, or

[1372] Administered at intervals of approximately 6 hours, or

[1373] Administered at intervals of approximately 5 hours, or

[1374] Administered at intervals of approximately 4 hours, or

[1375] Administered at intervals of approximately 3 hours; and / or

[1376] A beta-adrenergic receptor inhibitor is administered orally at a dose of about 30 mg, twice daily at intervals of about 7 to about 12 hours; and / or

[1377] Here:

[1378] The effective dose of an ADRB2 inhibitor is:

[1379] It is a pharmacologically effective dose with clinically acceptable serious side effects, or

[1380] About 10 mg to about 480 mg per day, or

[1381] About 20 mg to about 480 mg per day, or

[1382] About 30 mg to about 480 mg per day, or

[1383] About 40 mg to about 480 mg per day, or

[1384] About 50 mg to about 480 mg per day, or

[1385] About 60 mg to about 480 mg per day, or

[1386] About 70 mg to about 480 mg per day, or

[1387] About 80 mg to about 480 mg per day, or

[1388] About 120 mg to about 480 mg per day, or

[1389] About 160 mg to about 480 mg per day, or

[1390] About 200 mg to about 480 mg per day, or

[1391] About 240 mg to about 480 mg per day, or

[1392] About 280 mg to about 480 mg per day, or

[1393] About 320 mg to about 480 mg per day, or

[1394] About 360 mg to about 480 mg per day, or

[1395] About 400 mg to about 480 mg per day, or

[1396] About 440 mg to about 480 mg per day, or

[1397] About 10 mg to about 480 mg per day, or

[1398] About 10 mg to about 450 mg per day, or

[1399] It is about 10 mg to about 420 mg per day, or

[1400] About 10 mg to about 390 mg per day, or

[1401] About 10 mg to about 360 mg per day, or

[1402] About 10 mg to about 330 mg per day, or

[1403] About 10 mg to about 300 mg per day, or

[1404] About 10 mg to about 270 mg per day, or

[1405] About 10 mg to about 240 mg per day, or

[1406] About 10 mg to about 210 mg per day, or

[1407] It is about 10 mg to about 180 mg per day, or

[1408] About 10 mg to about 150 mg per day, or

[1409] It is about 10 mg to about 120 mg per day, or

[1410] It is about 10 mg to about 90 mg per day, or

[1411] It is about 10 mg to about 60 mg per day, or

[1412] It is about 10 mg to about 30 mg per day, or

[1413] It is about 10 mg to about 90 mg per day, or

[1414] It is about 10 mg to about 60 mg per day, or

[1415] It is about 10 mg to about 30 mg per day, or

[1416] About 20 mg to about 480 mg per day, or

[1417] About 20 mg to about 450 mg per day, or

[1418] About 20 mg to about 440 mg per day, or

[1419] About 20 mg to about 410 mg per day, or

[1420] About 20 mg to about 400 mg per day, or

[1421] About 20 mg to about 370 mg per day, or

[1422] About 20 mg to about 360 mg per day, or

[1423] About 20 mg to about 330 mg per day, or

[1424] About 20 mg to about 320 mg per day, or

[1425] About 20 mg to about 290 mg per day, or

[1426] About 20 mg to about 280 mg per day, or

[1427] About 20 mg to about 250 mg per day, or

[1428] About 20 mg to about 240 mg per day, or

[1429] About 20 mg to about 210 mg per day, or

[1430] About 20 mg to about 200 mg per day, or

[1431] About 20 mg to about 170 mg per day, or

[1432] About 20 mg to about 160 mg per day, or

[1433] About 20 mg to about 130 mg per day, or

[1434] About 20 mg to about 120 mg per day, or

[1435] It is about 20 mg to about 90 mg per day, or

[1436] It is about 20 mg to about 80 mg per day, or

[1437] It is about 20 mg to about 50 mg per day, or

[1438] It is about 20 mg to about 40 mg per day, or

[1439] About 20 mg to about 160 mg per day, or

[1440] About 20 mg to about 130 mg per day, or

[1441] About 20 mg to about 120 mg per day, or

[1442] About 20 mg to about 100 mg per day, or

[1443] It is about 20 mg to about 80 mg per day, or

[1444] It is about 20 mg to about 70 mg per day, or

[1445] It is about 20 mg to about 40 mg per day, or

[1446] About 20 mg per day, or

[1447] About 40 mg per day, or

[1448] Approximately 60 mg per day, or

[1449] Approximately 80 mg per day, and / or

[1450] ADRB2 inhibitors are:

[1451] Administered via a pharmacologically acceptable route, or

[1452] Administered orally; and / or

[1453] ADRB2 inhibitors are administered once daily, or twice daily, or three times daily, or four times daily; and / or

[1454] ADRB2 inhibitors are:

[1455] It is administered at intervals of approximately 7 to 12 hours, or

[1456] Administered at intervals of approximately 12 hours, or

[1457] Administered at intervals of approximately 11 hours, or

[1458] Administered at intervals of approximately 10 hours, or

[1459] Administered at intervals of approximately 9 hours, or

[1460] Administered at intervals of approximately 8 hours, or

[1461] Administered at intervals of approximately 7 hours, or

[1462] Administered at intervals of approximately 6 hours, or

[1463] Administered at intervals of approximately 5 hours, or

[1464] Administered at intervals of approximately 4 hours, or

[1465] Administered at intervals of approximately 3 hours; and / or

[1466] ADRB2 inhibitors are administered orally at a dose of about 30 mg, twice daily at intervals of about 7 to 12 hours; and / or

[1467] Here:

[1468] The effective amount of propranolol is:

[1469] About 10 mg to about 480 mg per day, or

[1470] About 20 mg to about 480 mg per day, or

[1471] About 30 mg to about 480 mg per day, or

[1472] About 40 mg to about 480 mg per day, or

[1473] About 50 mg to about 480 mg per day, or

[1474] About 60 mg to about 480 mg per day, or

[1475] About 70 mg to about 480 mg per day, or

[1476] About 80 mg to about 480 mg per day, or

[1477] About 120 mg to about 480 mg per day, or

[1478] About 160 mg to about 480 mg per day, or

[1479] About 200 mg to about 480 mg per day, or

[1480] About 240 mg to about 480 mg per day, or

[1481] About 280 mg to about 480 mg per day, or

[1482] About 320 mg to about 480 mg per day, or

[1483] About 360 mg to about 480 mg per day, or

[1484] About 400 mg to about 480 mg per day, or

[1485] About 440 mg to about 480 mg per day, or

[1486] About 10 mg to about 480 mg per day, or

[1487] About 10 mg to about 450 mg per day, or

[1488] It is about 10 mg to about 420 mg per day, or

[1489] About 10 mg to about 390 mg per day, or

[1490] About 10 mg to about 360 mg per day, or

[1491] About 10 mg to about 330 mg per day, or

[1492] About 10 mg to about 300 mg per day, or

[1493] About 10 mg to about 270 mg per day, or

[1494] About 10 mg to about 240 mg per day, or

[1495] About 10 mg to about 210 mg per day, or

[1496] It is about 10 mg to about 180 mg per day, or

[1497] About 10 mg to about 150 mg per day, or

[1498] It is about 10 mg to about 120 mg per day, or

[1499] It is about 10 mg to about 90 mg per day, or

[1500] It is about 10 mg to about 60 mg per day, or

[1501] It is about 10 mg to about 30 mg per day, or

[1502] It is about 10 mg to about 90 mg per day, or

[1503] It is about 10 mg to about 60 mg per day, or

[1504] It is about 10 mg to about 30 mg per day, or

[1505] About 20 mg to about 480 mg per day, or

[1506] About 20 mg to about 450 mg per day, or

[1507] About 20 mg to about 440 mg per day, or

[1508] About 20 mg to about 410 mg per day, or

[1509] About 20 mg to about 400 mg per day, or

[1510] About 20 mg to about 370 mg per day, or

[1511] About 20 mg to about 360 mg per day, or

[1512] About 20 mg to about 330 mg per day, or

[1513] About 20 mg to about 320 mg per day, or

[1514] About 20 mg to about 290 mg per day, or

[1515] About 20 mg to about 280 mg per day, or

[1516] About 20 mg to about 250 mg per day, or

[1517] About 20 mg to about 240 mg per day, or

[1518] About 20 mg to about 210 mg per day, or

[1519] About 20 mg to about 200 mg per day, or

[1520] About 20 mg to about 170 mg per day, or

[1521] About 20 mg to about 160 mg per day, or

[1522] About 20 mg to about 130 mg per day, or

[1523] About 20 mg to about 120 mg per day, or

[1524] It is about 20 mg to about 90 mg per day, or

[1525] It is about 20 mg to about 80 mg per day, or

[1526] It is about 20 mg to about 50 mg per day, or

[1527] It is about 20 mg to about 40 mg per day, or

[1528] About 20 mg to about 160 mg per day, or

[1529] About 20 mg to about 130 mg per day, or

[1530] About 20 mg to about 120 mg per day, or

[1531] About 20 mg to about 100 mg per day, or

[1532] It is about 20 mg to about 80 mg per day, or

[1533] It is about 20 mg to about 70 mg per day, or

[1534] It is about 20 mg to about 40 mg per day, or

[1535] About 20 mg per day, or

[1536] About 40 mg per day, or

[1537] Approximately 60 mg per day, or

[1538] Approximately 80 mg per day, and / or

[1539] Propranolol is:

[1540] Administered via a pharmacologically acceptable route, or

[1541] Administered orally; and / or

[1542] Propranolol is administered once daily, or twice daily, or three times daily, or four times daily; and / or

[1543] Propranolol is:

[1544] It is administered at intervals of approximately 7 to 12 hours, or

[1545] Administered at intervals of approximately 12 hours, or

[1546] Administered at intervals of approximately 11 hours, or

[1547] Administered at intervals of approximately 10 hours, or

[1548] Administered at intervals of approximately 9 hours, or

[1549] Administered at intervals of approximately 8 hours, or

[1550] Administered at intervals of approximately 7 hours, or

[1551] Administered at intervals of approximately 6 hours, or

[1552] Administered at intervals of approximately 5 hours, or

[1553] Administered at intervals of approximately 4 hours, or

[1554] Administered at intervals of approximately 3 hours; and / or

[1555] Propranolol is administered orally at a dose of about 30 mg, twice daily at intervals of about 7 to about 12 hours; and / or

[1556] Here:

[1557] The effective amount of G-CSF is:

[1558] Administered intravenously at about 1 µg / kg to about 70 µg / kg twice daily, or

[1559] Administered by subcutaneous injection at a dose of about 1 µg / kg to about 40 µg / kg per day, or

[1560] About 3 µg / kg to about 40 µg / kg per day by continuous subcutaneous injection, or

[1561] About 10 ug / kg to about 40 ug / kg per day, or

[1562] Approximately 10 ug / kg per day; and / or

[1563] G-CSF is:

[1564] Administered via a pharmacologically acceptable route, or

[1565] Administered intravenously, or

[1566] Administered subcutaneously, or

[1567] Administered continuously subcutaneously; and / or

[1568] The last dose of G-CSF inhibitor prior to the start of apheresis and / or leukocyte apheresis is:

[1569] When apheresis and / or leukocyte apheresis treatment begins

[1570] Administered about 120 hours to about 10 hours prior, or

[1571] Administered about 72 hours to about 10 hours prior, or

[1572] Administered about 24 hours to about 10 hours prior, or

[1573] Administered about 14 to 10 hours prior, or

[1574] Administered about 13 to 10 hours prior, or

[1575] Administered about 13 to 10 hours prior, or

[1576] Administered about 12 to 10 hours prior, or

[1577] Administered about 11 to 10 hours prior, or

[1578] Administered about 12 hours prior, or

[1579] Administered about 11 hours prior, or

[1580] Administered approximately 10 hours prior; and / or

[1581] G-CSF is administered once daily; and / or

[1582] G-CSF is administered 1, 2, 3, 4, 5, 6, 7, 8, or more than 8 times; and / or

[1583] G-CSF is administered 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, or 3 times or less, 2 times or less, or 1 time or less; and / or

[1584] A method in which G-CSF is administered by intravenous injection, subcutaneous injection, or serial subcutaneous injection at a rate of about 10 µg / kg per day for about 4 days, preferably at least about 10 hours prior to apheresis and / or leukocyte apheresis.

[1585] Example 116. In any one of the embodiments 98 to the immediately above,

[1586] CD34 in the subject + A drug that interferes with cell mobilization is administered,

[1587] The drug is lenalidomide, and

[1588] Lenalidomide is:

[1589] Administered according to the drug label, and / or

[1590] Administered at a dose of about 8 mg / kg to about 16 mg / kg, preferably about 16 mg / kg, and / or

[1591] Administered via a pharmacologically acceptable route, or

[1592] Administered intravenously, or

[1593] Administered for about 3 to about 4 cycles; and / or

[1594] The drug is daratumumab, and

[1595] Daratumumab is:

[1596] Administered according to the drug label, and / or

[1597] Administered at a dose of about 8 mg / kg to about 16 mg / kg, preferably about 16 mg / kg, and / or

[1598] Administered via a pharmacologically acceptable route, or

[1599] Administered intravenously, or

[1600] A method administered for about 3 to 4 cycles.

[1601] Example 117. In any one of the embodiments 98 to the immediately above,

[1602] The method is:

[1603] Does not cause adverse reactions in subjects during or after treatment, or

[1604] Does not cause severe adverse reactions in subjects during or after treatment, or

[1605] Does not cause adverse reactions of grade 1 or 2 or greater severity, or

[1606] A method in which adverse reactions, if caused, are limited to a list consisting of bone pain, hypocalcemia, diarrhea, dry mouth, nausea, hypokalemia, peripheral edema, paresthesia, etc.

Claims

Claim 1 A method for mobilizing cells in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. Claim 2 A method for inducing cell recruitment in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. Claim 3 A method for enhancing apheresis by inducing cell mobilization in a subject, comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in a subject. Claim 4 A method for improving apheresis by mobilizing cells from a subject, the method comprising the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. Claim 5 A method in which, in any one of paragraphs 1 to 4, the cell is a stem cell. Claim 6 A method according to any one of claims 1 to 5, wherein the cell is an LSK(Lin-Sca1+c-Kit+) cell. Claim 7 In claim 6, the LSK cells are long-term repopulating hematopoietic stem cells (LT-HSC), method. Claim 8 In claim 7, the LT-HSC is a CD150+, CD34-, or CD48+ LSK cell, method. Claim 9 A method according to any one of claims 1 to 8, wherein the step of blocking CXCR4 signaling includes the step of administering a CXCR4 inhibitor to a subject. Claim 10 In claim 9, the CXCR4 inhibitor forms an ion interaction with Arg188 on CXCR4, a method. Claim 11 The method of claim 9 or 10, wherein the CXCR4 inhibitor forms a hydrogen bond with Gln200 on CXCR4. Claim 12 A method according to any one of claims 9 to 11, wherein the CXCR4 inhibitor has a higher binding affinity for CXCL12 than AMD3100. Claim 13 A method according to any one of claims 9 to 12, wherein the CXCR4 inhibitor has an inhibitory constant (Ki) lower than that of AMD3100. Claim 14 A method according to any one of claims 9 to 13, wherein the CXCR4 inhibitor has an inhibition constant at least twice lower than that of AMD3100. Claim 15 A method according to any one of claims 9 to 14, wherein the CXCR4 inhibitor inhibits CXCL12-induced calcium flux to a level equivalent to or higher than that of AMD3100. Claim 16 A method according to any one of claims 9 to 15, wherein the CXCR4 inhibitor inhibits CXCL12-induced arrestin recruitment to a level equivalent to or higher than that of AMD3100. Claim 17 A method according to any one of claims 9 to 16, wherein the CXCR4 inhibitor inhibits CXCL12-induced migration of cancer cells to a level equivalent to or higher than that of AMD3100. Claim 18 A method according to any one of claims 9 to 17, wherein the CXCR4 inhibitor inhibits CXCL12-induced migration of U937 and MM.1S cells to a level equivalent to or higher than that of AMD3100. Claim 19 A method according to any one of claims 9 to 18, wherein the CXCR4 inhibitor inhibits calcium flux induced by CXCL12 and epinephrine. Claim 20 A method according to any one of claims 9 to 19, wherein blocking beta-adrenergic receptor signaling inhibits calcium flux and / or cell migration induced by CXCL12 and epinephrine. Claim 21 A method according to any one of claims 9 to 19, wherein the blocking of propranolol inhibits calcium flux and / or cell migration induced by CXCL12 and epinephrine. Claim 22 A method according to any one of claims 1 to 21, wherein the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in a subject induces an enhanced amount of long-term reconstituted hematopoietic stem cell (LT-HSC) recruitment compared to the amount of LT-HSC recruitment induced by a CXCR4 inhibitor and G-CSF alone. Claim 23 A method according to any one of claims 1 to 22, wherein the step of blocking CXCR4 signaling and beta-adrenergic receptor signaling in a subject recruits LT-HSCs in an increased amount compared to the amount of LT-HSC recruitment induced by a CXCR4 inhibitor and G-CSF alone. Claim 24 In paragraph 22 or 23, the LT-HSC is a method that expresses CD150. Claim 25 In paragraph 22 or 23, the method wherein LT-HSC does not express CD34. Claim 26 In paragraph 22 or 23, the LT-HSC is a method that expresses CD48. Claim 27 A method comprising, in any one of claims 1 to 26, an additional step of administering G-CSF. Claim 28 In claim 27, a method wherein a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induce an increased amount of cell mobilization compared to the amount of cell mobilization induced by a CXCR4 inhibitor and G-CSF alone. Claim 29 In claim 27, a method wherein a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF mobilize cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor and G-CSF alone. Claim 30 A method according to any one of claims 1 to 29, wherein the step of blocking beta-adrenergic receptor signaling is performed before the step of blocking CXCR4 signaling. Claim 31 A method according to any one of claims 1 to 30, wherein the step of blocking beta-adrenergic receptor signaling continues even after the step of blocking CXCR4 signaling is terminated. Claim 32 A method according to any one of claims 1 to 31, wherein the step of blocking CXCR4 signaling includes the step of administering a CXCR4 inhibitor to a subject. Claim 33 A method according to any one of claims 1 to 32, wherein the step of blocking beta-adrenergic receptor signaling includes the step of administering a beta-adrenergic receptor inhibitor to a subject. Claim 34 In paragraph 33, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor, method. Claim 35 In paragraph 33 or 34, the beta-adrenergic receptor inhibitor comprises alprenolol, atenolol, betaxolol, bupranolol, butoxamine, carazolol, carvedilol, CGP 12177, cicloprolol, ICI 118551, ICYP, labetalol, levobetaxolol, levobunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, and timolol. A method selected from the military. Claim 36 In paragraph 35, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551. Claim 37 In paragraph 36, the beta-adrenergic receptor inhibitor is propranolol, method. Claim 38 In any one of claims 9 to 37, the CXCR4 inhibitor is ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549, D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GSTNT21MP, Isothiourea-1a, Isothiourea-1t (IT1t), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[11C]methyl-AMD3465, POL6326, SDF-1 l-9[P2G] dimer, SDFl P2G, T134, T140, T22, TC 14012, GPC-100 (Bulixafor, or TG-0054), USL311, Viral macrophage inflammation protein-II (vMIP-11), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]O2-AMD3100, [177Lu]pentixather, Ulocuplumab A method selected from the group consisting of (MDX1338 / BMS-936564), AD-214, LY2624587, PF-06747143, POL6326, MB1707, and 508MC1 (compound 26). Claim 39 In claim 38, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozovil), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, GPC-100 (Bulixafor, or TG-0054), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). Claim 40 In paragraph 39, the method wherein the CXCR4 inhibitor is GPC-100 (burixafor, or TG-0054). Claim 41 In paragraph 39, the CXCR4 inhibitor is AMD3100 (Plerixafor, or Mozobil), method. Claim 42 In paragraph 39, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564), method. Claim 43 A method according to any one of claims 33 to 42, wherein the step of administering a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor to a subject comprises the step of administering GPC-100 (burixafor, or TG-0054) and propranolol. Claim 44 A method according to any one of claims 33 to 42, wherein the step of administering a CXCR4 inhibitor beta-adrenergic receptor inhibitor to a subject comprises the step of administering AMD3100 (plerixafor, or mozobil) and propranolol. Claim 45 A method according to any one of claims 33 to 42, wherein the step of administering a CXCR4 inhibitor beta-adrenergic receptor inhibitor to a subject comprises the step of administering ulocuplumab (MDX1338 / BMS-936564) and propranolol. Claim 46 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone. Claim 47 A method according to any one of claims 27 to 42, wherein the step of administering the combination of a CXCR4 inhibitor and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone. Claim 48 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor alone. Claim 49 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by a CXCR4 inhibitor alone. Claim 50 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor alone. Claim 51 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor alone. Claim 52 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor alone. Claim 53 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor alone. Claim 54 A method according to any one of claims 27 to 42, wherein the step of administering a combination of a CXCR4 inhibitor, a beta-adrenergic receptor inhibitor, and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by a CXCR4 inhibitor and a beta-adrenergic receptor inhibitor alone. Claim 55 A method according to any one of claims 27 to 42, wherein the step of administering a combination of GPC-100 (burixafor, or TG-0054) and G-CSF induces an increased amount of cell mobilization compared to the amount of cell mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF. Claim 56 A method according to any one of claims 27 to 42, wherein the step of administering a combination of GPC-100 (burixafor, or TG-0054) and G-CSF mobilizes cells in an increased amount compared to the amount of cell mobilization induced by AMD3100 (plerixafor, or mozobil) and G-CSF. Claim 57 A method according to any one of claims 27 to 42, wherein the step of administering a combination of GPC-100 (burixafor, or TG-0054) and G-CSF induces an increased amount of apheresis compared to the amount of apheresis induced by AMD3100 (plerixafor, or mozobil) and G-CSF. Claim 58 A method according to any one of claims 47 to 57, wherein the increased amount of cell mobilization or component apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) analysis. Claim 59 In paragraph 58, the increased amount of cell mobilization or component collection is measured by flow cytometry. Claim 60 In paragraph 59, the method wherein flow cytometry is performed on (Lin-Scal+c-Kit+) LSK cells. Claim 61 In paragraph 58, the increased amount of cell mobilization or component collection is measured by colony-forming unit (CFU) analysis. Claim 62 A method according to any one of claims 1 to 61, wherein the subject has a CXCR4 protomer in the cell. Claim 63 A method according to any one of claims 1 to 62, wherein the subject has an ADRB2 protomer in a cell. Claim 64 A method according to any one of claims 1 to 63, wherein the subject has a CXCR4 protomer and an ADRB2 protomer within the cell. Claim 65 In paragraph 64, the subject has a CXCR4-ADRB2 heteromer in the cell, a method. Claim 66 A method according to claim 65, wherein i) the CXCR4-ADRB2 heteromer has an increased amount of downstream calcium mobilization compared to downstream calcium mobilization from the CXCR4 protomer or ADRB2 protomer; and ii) the administered inhibitor combination inhibits the enhanced downstream calcium mobilization from the CXCR4-ADRB2 heteromer within the stem cell. Claim 67 A method according to any one of claims 5 to 66, wherein the stem cell is selected from the group consisting of hematopoietic stem cells, hematopoietic progenitor cells, mesenchymal stem cells, endothelial progenitor cells, neural stem cells, epithelial stem cells, skin stem cells, and cancer stem cells. Claim 68 In paragraph 67, the method wherein the stem cells are hematopoietic stem cells or hematopoietic progenitor cells. Claim 69 In paragraph 68, a method in which hematopoietic stem cells or hematopoietic progenitor cells are mobilized from bone marrow to peripheral blood (PB). Claim 70 In paragraph 69, the method of collecting mobilized hematopoietic stem cells or hematopoietic progenitor cells for transplantation into a patient with cancer. Claim 71 In paragraph 70, the cancer is selected from the group consisting of lymphoma, leukemia, and multiple myeloma. Claim 72 In paragraph 71, the cancer is non-Hodgkin's lymphoma (NHL), Hodgkin's Disease (HD) or Hodgkin's Lymphoma (HL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), multiple myeloma (MM) or Burkitt's Lymphoma. Claim 73 A method in which, in any one of paragraphs 5 through 66, the stem cell is a mesenchymal stem cell. Claim 74 In paragraph 73, a method in which mesenchymal stem cells are mobilized from bone marrow to peripheral blood. Claim 75 In claim 74, a method wherein mesenchymal stem cells are mobilized for the treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. Claim 76 A method in which, in any one of paragraphs 5 through 66, the stem cells are cancer stem cells. Claim 77 In paragraph 76, a method of mobilizing cancer stem cells into the blood. Claim 78 In paragraph 76 or 77, a method in which cancer stem cells are mobilized for the treatment of cancer. Claim 79 A method in which, in any one of paragraphs 5 through 66, the cell is an immune cell. Claim 80 In paragraph 79, the immune cells are white blood cells (WBC). Claim 81 In paragraph 80, the method is that WBC is a lymphocyte. Claim 82 In paragraph 81, the lymphocytes are selected from the group consisting of T cells, B cells, and natural killer (NK) cells. Claim 83 In paragraph 82, the lymphocyte is a T cell, method. Claim 84 In paragraph 82, the lymphocytes are natural killer (NK) cells. Claim 85 In paragraph 80, the WBC is a granulocyte, method. Claim 86 In paragraph 85, the granulocytes are selected from the group consisting of neutrophils, eosinophils, and basophils. Claim 87 In paragraph 86, the granulocyte is a neutrophil, method. Claim 88 In paragraph 81, WBC is a monocyte, method. Claim 89 A method in which, in any one of paragraphs 79 to 88, immune cells are mobilized from the bone marrow to the peripheral blood. Claim 90 A method in which, in any one of paragraphs 79 to 88, immune cells are mobilized from the spleen to peripheral blood. Claim 91 A method in which, in any one of paragraphs 79 to 88, immune cells are mobilized from the thymus to peripheral blood. Claim 92 A method in which, in any one of paragraphs 79 to 88, immune cells are mobilized from lymph nodes to peripheral blood. Claim 93 A method according to any one of paragraphs 79 through 88, wherein the mobilized immune cells are used in adoptive cell therapy (ACT). Claim 94 In paragraph 93, the adoptive cell therapy (ACT) is a chimeric antigen receptor (CAR) T cell therapy, a method. Claim 95 In paragraph 93, the adoptive cell therapy (ACT) is a method that is a natural killer (NK) cell therapy. Claim 96 In paragraph 93, the adoptive cell therapy (ACT) is a method that is engineered T-cell receptor (TCR) therapy. Claim 97 In paragraph 93, the adoptive cell therapy (ACT) is a tumor-infiltrating lymphocyte (TIL) therapy, a method. Claim 98 As a method of preparing a subject for treatment, the method comprises the following: Step of blocking CXCR4 in the target, A step of blocking beta-adrenergic receptors in a subject, or Step of blocking CXCR4 inhibitors and beta-adrenergic receptors in a subject. Claim 99 In paragraph 98, Blocking of CXCR4 is achieved through ionic interaction with Arg188 on CXCR4; and / or Blocking of CXCR4 is achieved through hydrogen bonding with Gln200 on CXCR4; and / or any combination thereof, method. Claim 100 As a treatment method for a subject in need, the method comprises the following: Step of blocking CXCR4 in the target, A step of blocking beta-adrenergic receptors in a subject, or Step of blocking CXCR4 inhibitors and beta-adrenergic receptors in a subject. Claim 101 In Article 100, Blocking of CXCR4 is achieved through ionic interaction with Arg188 on CXCR4; and / or Blocking of CXCR4 is achieved through hydrogen bonding with Gln200 on CXCR4; and / or any combination thereof, method. Claim 102 A method for preparing a subject for treatment, comprising the step of administering the following effective amount: CXCR4 inhibitor, Beta-adrenergic receptor inhibitors, or Their combination. Claim 103 A method for qualifying a subject for treatment, comprising the step of administering the following effective amount: CXCR4 inhibitor, Beta-adrenergic receptor inhibitors, or Their combination. Claim 104 A method for treating a subject in need, comprising the step of administering the following effective amount: CXCR4 inhibitor, Beta-adrenergic receptor inhibitors, or Their combination. Claim 105 Use of GPC-100 for the manufacture of a drug for the treatment of cancer, or neurological disorders, or any combination thereof. Claim 106 A method according to any one of claims 98 to 105, wherein the treatment further comprises the step of administering the following effective amount: Beta-adrenergic receptor inhibitors, or G-CSF, or lenalidomide, or daratumumab, or Velacade, or Dexamethasone, or cyclophosphamide, or Bortezombil, or Their combination. Claim 107 A method according to any one of claims 98 to 106, wherein the subject is pretreated by administering the following effective amount: Beta-adrenergic receptor inhibitors, or G-CSF, or Lenalidomide, or Daratumumab, or Velacade, or Dexamethasone, or Cyclophosphamide, or Bortezomib, or Their combination. Claim 108 In any one of paragraphs 98 through 107, the treatment comprises one or more of the following methods: apheresis step; or leukapheresis step; or Peripheral blood collection step; or As a cell transplantation step, The cells are selectively stem cells, and The cells are selectively hematopoietic progenitor cells (HSCs), and The cell is selectively a T cell, or a combination thereof, step; and / or CAR-T therapy stage; or Their combination. Claim 109 A method comprising, in any one of claims 98 to 108, further comprising the step of administering an effective amount of G-CSF. Claim 110 In any one of paragraphs 98 through 109, The subject exhibits or is diagnosed with symptoms of cancer, or neurological disorders, or any combination thereof; and / or Cancer is selected from the group consisting of lymphoma, leukemia, and multiple myeloma; and / or The subject exhibits symptoms of or is diagnosed with hematologic malignancy; and / or The cancer is selected from the group consisting of non-Hodgkin lymphoma (NHL), Hodgkin's disease (HD) or Hodgkin lymphoma (HL), acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), multiple myeloma (MM), and Burkitt lymphoma; and / or The cancer is HD; and / or The arm is NHL and; and / or The cancer is AML; and / or Cancer is ALL and; and / or The cancer is MM; and / or Neurological disorders are selected from the group consisting of neurological disorders, cardiac ischemia, myocardial infarction, diabetes mellitus, tissue repair, bone and cartilage diseases, autoimmune diseases, graft-versus-host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis; and / or The subject is a human, method. Claim 111 In any one of paragraphs 98 through 110, The subject requires an Autologous Stem Cell Transplant (ASCT); The treatment is a method called autologous stem cell transplantation (ASCT). Claim 112 In any one of paragraphs 98 through 111, Here: CXCR4 inhibitors form ion interactions with Arg188 on CXCR4, or CXCR4 inhibitors form hydrogen bonds with Gln200 on CXCR4, or Any combination of these; and / or Here: CXCR4 inhibitors have a higher binding affinity for CXCL12 than AMD3100, or CXCR4 inhibitors have a lower inhibition constant (Ki) than AMD3100, or A CXCR4 inhibitor having an inhibition constant at least twice lower than AMD3100, method. Claim 113 In any one of paragraphs 98 through 112, CXCR4 inhibitors are: ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (Plericsaphor, or Mozobil), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549, D-[Lys3]GHRP-6, FC122, FC131, GMI-1359, GSK812397, GSTNT21MP, Isothiourea-1a, Isothiourea-1t (IT1t), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[11C]methyl-AMD3465, POL6326, SDF-1 l-9[P2G] dimer, SDFl P2G, T134, T140, T22, TC 14012, GPC-100 (burixafor, or TG-0054), USL311, viral macrophage inflammation protein-II (vMIP-11), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixater, [99mTc]O2-AMD3100, [177Lu]pentixater, ulocuplumab (MDX1338 / BMS-936564), AD-214, LY2624587, PF-06747143, POL6326. Selected from the group consisting of MB1707, 508MC1 (Compound 26), and derivatives and similars thereof; and / or Selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (Plerixafor, or Mozobil), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, GPC-100 (Bulixafor, or TG-0054), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564), and derivatives and analogs thereof; and / or ulocuflumab (MDX1338 / BMS-936564) and / or AMD3100 (Pleric Sapor, or Mojobil) and / or BL8040 and; and / or GPC-100 (Buriksapor, or TG-0054) and; and / or Beta-adrenergic receptor inhibitors are: It is an ADRB2 inhibitor, and / or Selected from the group consisting of alfrenollol, atenolol, betaxolol, bufranolol, butoxamine, carazolol, carvedilol, CGP 12177, cycloprollol, ICI 118551, ICYP, labetalol, levobetaxolol, levovunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, timolol, and derivatives and analogs thereof, and / or Selected from the group consisting of propranolol, nadolol, and ICI 118551, and / or Propranolol, method. Claim 114 In Paragraph 113, The CXCR4 inhibitor is GPC-100, or AMD3100, or BL8040, preferably GPC-100; and / or The beta-adrenergic receptor inhibitor is propranolol, method. Claim 115 In any one of paragraphs 98 through 114, Here: The effective dose of the CXCR inhibitor is: It is a pharmacologically effective dose with clinically acceptable serious side effects, or It is about 30 mg / kg to about 0.01 mg / kg, or It is about 30 mg / kg to about 0.07 mg / kg, or It is about 25 mg / kg to about 0.07 mg / kg, or It is about 20 mg / kg to about 0.07 mg / kg, or It is about 15 mg / kg to about 0.07 mg / kg, or It is about 10 mg / kg to about 0.07 mg / kg, or About 9 mg / kg to about 0.07 mg / kg, or It is about 8 mg / kg to about 0.07 mg / kg, or It is about 7 mg / kg to about 0.07 mg / kg, or It is about 6 mg / kg to about 0.07 mg / kg, or It is about 5 mg / kg to about 0.07 mg / kg, or About 4.40 mg / kg to about 0.07 mg / kg, or About 3.14 mg / kg to about 0.07 mg / kg, or About 2.24 mg / kg to about 0.07 mg / kg, or About 1.12 mg / kg to about 0.07 mg / kg, or About 0.56 mg / kg to about 0.07 mg / kg, or About 0.28 mg / kg to about 0.07 mg / kg, or About 0.14 mg / kg to about 0.07 mg / kg, or About 0.10 mg / kg to about 0.07 mg / kg, or It is about 30 mg / kg to about 0.10 mg / kg, or It is about 25 mg / kg to about 0.10 mg / kg, or It is about 20 mg / kg to about 0.10 mg / kg, or It is about 15 mg / kg to about 0.10 mg / kg, or It is about 10 mg / kg to about 0.10 mg / kg, or It is about 9 mg / kg to about 0.10 mg / kg, or It is about 8 mg / kg to about 0.10 mg / kg, or It is about 7 mg / kg to about 0.10 mg / kg, or It is about 6 mg / kg to about 0.10 mg / kg, or It is about 5 mg / kg to about 0.10 mg / kg, or It is about 4.40 mg / kg to about 0.10 mg / kg, or About 3.14 mg / kg to about 0.10 mg / kg, or About 2.24 mg / kg to about 0.10 mg / kg, or About 1.12 mg / kg to about 0.10 mg / kg, or About 0.56 mg / kg to about 0.10 mg / kg, or About 0.28 mg / kg to about 0.10 mg / kg, or About 0.14 mg / kg to about 0.10 mg / kg, or Approximately 4.40 mg / kg, or Approximately 3.14 mg / kg, or Approximately 2.24 mg / kg, or Approximately 1.12 mg / kg, or Approximately 0.56 mg / kg, or Approximately 0.28 mg / kg, or Approximately 0.14 mg / kg, or Approximately 0.10 mg / kg, or Approximately 0.07 mg / kg, or CXCR4 inhibitors are: Administered via a pharmacologically acceptable route, or Administered orally, or Administered subcutaneously, or Administered intravenously; and / or The duration of CXCR4 inhibitor administration is: It is about 1 minute to about 30 minutes, or It is about 5 minutes to about 20 minutes, or It is about 10 minutes to about 18 minutes, or It is about 15 minutes and; and / or The time of administration for CXCR4 inhibitors is: It is morning, or It is about 7:00 AM to about 7 PM, or It is around 7:30 AM to around 12:30 PM, or It is around 7:30 AM to around 9:30 PM, or It is about 8:30 AM and; and / or The last dose of CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is: When apheresis and / or leukocyte apheresis treatment begins Administered about 120 hours to about 30 minutes prior, or Administered about 72 hours to about 30 minutes prior, or Administered about 24 hours to about 30 minutes prior, or Administered about 14 hours to about 30 minutes prior, or Administered about 13 hours to about 30 minutes prior, or Administered about 13 hours to about 30 minutes prior, or Administered about 12 hours to about 30 minutes prior, or Administered about 11 hours to about 30 minutes prior, or Administered about 10 hours to about 30 minutes prior, or Administered about 9 hours to about 30 minutes prior, or Administered about 8 hours to about 30 minutes prior, or Administered about 7 hours to about 30 minutes prior, or Administered about 6 hours to about 30 minutes prior, or Administered about 5 hours to about 30 minutes prior, or Administered about 4 hours to about 30 minutes prior, or Administered about 3 hours to about 30 minutes prior, or Administered about 2 hours to about 30 minutes prior, or Administered about 90 minutes to about 30 minutes prior, or Administered about 60 minutes to about 30 minutes prior, or Administered about 45 minutes to about 30 minutes prior, or Administered about 11 hours prior, or Administered about 10 hours prior, or Administered about 9 hours prior, or Administered about 8 hours prior, or Administered about 7 hours prior, or Administered about 6 hours prior, or Administered about 5 hours prior, or Administered about 4 hours prior, or Administered about 3 hours prior, or Administered about 2 hours prior, or Administered about 90 minutes prior, or Administered about 60 minutes prior, or Administered about 45 minutes prior, or Administered about 30 minutes prior; and / or CXCR4 inhibitors are administered once, twice, or three, four, or more than four times; and / or CXCR4 inhibitors are administered one or fewer times, two or fewer times, three or fewer times, four or fewer times, or five or fewer times; and / or CXCR4 inhibitors are administered for 1, 2, or 3, or 4, or for more than 4 days; and / or CXCR4 inhibitors are administered for 1 day or less, 2 days or less, 3 days or less, 4 days or less, or 5 days or less; and / or The apheresis and / or leukocyte apheresis treatment comprises about 1 to about 5 sessions, preferably about 1 to 2 sessions, more preferably 1 session; and / or CXCR4 inhibitors are administered once daily; and / or CXCR4 inhibitors are: It is administered about 7 to about 11 days after the administration of propranolol, or It is administered about 9 to 11 days after the administration of propranolol, or Administered about 7 to 8 days after administration of propranolol; or Administered on the 7th day after administration of propranolol, or Administered on the 8th day after administration of propranolol; and / or The first dose of the CXCR4 inhibitor is: It is administered about 7 to about 11 days after the administration of propranolol, or It is administered about 7 to 8 days after the administration of propranolol, or Administered on the 7th day after administration of propranolol, or Administered on the 8th day after administration of propranolol; and / or The second dose of the CXCR4 inhibitor is: It is administered about 7 to about 11 days after the administration of propranolol, or It is administered about 9 to 11 days after the administration of propranolol, or Administered on the 9th day after administration of propranolol, or Administered on the 10th day after administration of propranolol, or Administered on the 11th day after administration of propranolol; and / or About 0.1 mg / kg to about 4.4 mg / kg of a CXCR4 inhibitor is administered intravenously once daily for 1 day or up to 5 days, preferably the last dose of the CXCR4 inhibitor prior to apheresis and / or leukocyte apheresis is administered about 30 to about 60 minutes before the start of apheresis and / or leukocyte apheresis treatment; and / or Here: The effective amount of GPC-100 is: It is about 30 mg / kg to about 0.07 mg / kg, or It is about 25 mg / kg to about 0.07 mg / kg, or It is about 20 mg / kg to about 0.07 mg / kg, or It is about 15 mg / kg to about 0.07 mg / kg, or It is about 10 mg / kg to about 0.07 mg / kg, or About 9 mg / kg to about 0.07 mg / kg, or It is about 8 mg / kg to about 0.07 mg / kg, or It is about 7 mg / kg to about 0.07 mg / kg, or It is about 6 mg / kg to about 0.07 mg / kg, or It is about 5 mg / kg to about 0.07 mg / kg, or About 4.40 mg / kg to about 0.07 mg / kg, or About 3.14 mg / kg to about 0.07 mg / kg, or About 2.24 mg / kg to about 0.07 mg / kg, or About 1.12 mg / kg to about 0.07 mg / kg, or About 0.56 mg / kg to about 0.07 mg / kg, or About 0.28 mg / kg to about 0.07 mg / kg, or About 0.14 mg / kg to about 0.07 mg / kg, or About 0.10 mg / kg to about 0.07 mg / kg, or It is about 30 mg / kg to about 0.10 mg / kg, or It is about 25 mg / kg to about 0.10 mg / kg, or It is about 20 mg / kg to about 0.10 mg / kg, or It is about 15 mg / kg to about 0.10 mg / kg, or It is about 10 mg / kg to about 0.10 mg / kg, or It is about 9 mg / kg to about 0.10 mg / kg, or It is about 8 mg / kg to about 0.10 mg / kg, or It is about 7 mg / kg to about 0.10 mg / kg, or It is about 6 mg / kg to about 0.10 mg / kg, or It is about 5 mg / kg to about 0.10 mg / kg, or It is about 4.40 mg / kg to about 0.10 mg / kg, or About 3.14 mg / kg to about 0.10 mg / kg, or About 2.24 mg / kg to about 0.10 mg / kg, or About 1.12 mg / kg to about 0.10 mg / kg, or About 0.56 mg / kg to about 0.10 mg / kg, or About 0.28 mg / kg to about 0.10 mg / kg, or About 0.14 mg / kg to about 0.10 mg / kg, or Approximately 4.40 mg / kg, or Approximately 3.14 mg / kg, or Approximately 2.24 mg / kg, or Approximately 1.12 mg / kg, or Approximately 0.56 mg / kg, or Approximately 0.28 mg / kg, or Approximately 0.14 mg / kg, or Approximately 0.10 mg / kg, or Approximately 0.07 mg / kg, or GPC-100 is: Administered via a pharmacologically acceptable route, or Administered orally, or Administered subcutaneously, or Administered intravenously; and / or The duration of GPC-100 administration is: It is about 1 minute to about 30 minutes, or It is about 5 minutes to about 20 minutes, or It is about 10 minutes to about 18 minutes, or It is about 15 minutes and; and / or The administration time of GPC-100 is: It is morning, or It is about 7:00 AM to about 7 PM, or It is around 7:30 AM to around 12:30 PM, or It is around 7:30 AM to around 9:30 PM, or It is about 8:30 AM and; and / or The last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is: When apheresis and / or leukocyte apheresis treatment begins Administered about 120 hours to about 30 minutes prior, or Administered about 72 hours to about 30 minutes prior, or Administered about 24 hours to about 30 minutes prior, or Administered about 14 hours to about 30 minutes prior, or Administered about 13 hours to about 30 minutes prior, or Administered about 13 hours to about 30 minutes prior, or Administered about 12 hours to about 30 minutes prior, or Administered about 11 hours to about 30 minutes prior, or Administered about 10 hours to about 30 minutes prior, or Administered about 9 hours to about 30 minutes prior, or Administered about 8 hours to about 30 minutes prior, or Administered about 7 hours to about 30 minutes prior, or Administered about 6 hours to about 30 minutes prior, or Administered about 5 hours to about 30 minutes prior, or Administered about 4 hours to about 30 minutes prior, or Administered about 3 hours to about 30 minutes prior, or Administered about 2 hours to about 30 minutes prior, or Administered about 90 minutes to about 30 minutes prior, or Administered about 60 minutes to about 30 minutes prior, or Administered about 45 minutes to about 30 minutes prior, or Administered about 11 hours prior, or Administered about 10 hours prior, or Administered about 9 hours prior, or Administered about 8 hours prior, or Administered about 7 hours prior, or Administered about 6 hours prior, or Administered about 5 hours prior, or Administered about 4 hours prior, or Administered about 3 hours prior, or Administered about 2 hours prior, or Administered about 90 minutes prior, or Administered about 60 minutes prior, or Administered about 45 minutes prior, or Administered about 30 minutes prior; and / or GPC-100 is administered 1, 2, or 3, 4, or more than 4 times; and / or GPC-100 is administered 1 or fewer times, 2 or fewer times, 3 or fewer times, 4 or fewer times, or 5 or fewer times; and / or GPC-100 is administered for 1, 2, or 3, or 4, or for more than 4 days; and / or GPC-100 is administered for 1 day or less, 2 days or less, 3 days or less, 4 days or less, or 5 days or less; and / or The apheresis and / or leukocyte apheresis treatment comprises about 1 to about 5 sessions, preferably about 1 to 2 sessions, more preferably 1 session; and / or GPC-100 is administered once daily; and / or GPC-100 is: It is administered about 7 to about 11 days after the administration of propranolol, or It is administered about 9 to 11 days after the administration of propranolol, or Administered about 7 to 8 days after administration of propranolol; or Administered on the 7th day after administration of propranolol, or Administered on the 8th day after administration of propranolol; and / or The first dose of GPC-100 is: It is administered about 7 to about 11 days after the administration of propranolol, or It is administered about 7 to 8 days after the administration of propranolol, or Administered on the 7th day after administration of propranolol, or Administered on the 8th day after administration of propranolol; and / or The second dose of GPC-100 is: It is administered about 7 to about 11 days after the administration of propranolol, or It is administered about 9 to 11 days after the administration of propranolol, or Administered on the 9th day after administration of propranolol, or Administered on the 10th day after administration of propranolol, or Administered on the 11th day after administration of propranolol; and / or GPC-100 at a dose of about 0.1 mg / kg to about 4.4 mg / kg is administered intravenously once daily for 1 day or up to 5 days, preferably the last dose of GPC-100 prior to apheresis and / or leukocyte apheresis is administered about 30 to about 60 minutes before the start of apheresis and / or leukocyte apheresis treatment; and / or Here: The effective capacity of the AMD3100 is: It is about 10 μg / kg to about 480 μg / kg, or About 40 μg / kg to about 480 μg / kg, or About 80 μg / kg to about 480 μg / kg, or It is about 40 μg / kg to about 480 μg / kg and is administered subcutaneously; or It is about 80 μg / kg to about 480 μg / kg and is administered subcutaneously; or It is approximately 40 μg / kg and is administered subcutaneously; or It is approximately 80 μg / kg and is administered subcutaneously; or It is approximately 480 μg / kg and is administered subcutaneously; or It is about 10 μg / kg to about 80 μg / kg and is administered intravenously; or It is about 20 μg / kg to about 80 μg / kg and is administered intravenously; or It is about 40 μg / kg to about 80 μg / kg and is administered intravenously; or It is approximately 10 μg / kg and is administered intravenously; or It is approximately 20 μg / kg and is administered intravenously; or It is approximately 40 μg / kg and is administered intravenously; or It is approximately 80 μg / kg and is administered intravenously; and / or AMD3100 is: Administered via a pharmacologically acceptable route, or Administered subcutaneously, or Administered intravenously; and / or The last dose of the AMD3100 inhibitor prior to the start of apheresis and / or leukocyte apheresis is: When apheresis and / or leukocyte apheresis treatment begins Administered about 120 hours to about 10 hours prior, or Administered about 72 hours to about 10 hours prior, or Administered about 24 hours to about 10 hours prior, or Administered about 14 to 10 hours prior, or Administered about 13 to 10 hours prior, or Administered about 13 to 10 hours prior, or Administered about 12 to 10 hours prior, or Administered about 11 to 10 hours prior, or Administered about 12 hours prior, or Administered about 11 hours prior, or Administered approximately 10 hours prior; and / or AMD3100 is administered once daily; and / or AMD3100 is administered 1, 2, 3, 4, or more than 4 times; and / or AMD3100 is administered 5 times or less, 4 times or less, or 3 times or less, 2 times or less, or 1 time or less; and / or AMD3100 is administered intravenously or subcutaneously, preferably subcutaneously, at a dose of about 10 μg / kg to about 480 μg / kg mg / kg once daily for 1 day or up to 5 days, preferably the last dose of AMD3100 prior to apheresis and / or leukocyte apheresis is administered at least about 10 hours prior to the initiation of apheresis and / or leukocyte apheresis treatment, or preferably about 12 to about 10 hours prior to the initiation of apheresis and / or leukocyte apheresis treatment; and / or Here: The effective amount of BL8040 is: About 0.5 mg / kg to about 2.0 mg / kg, or About 0.75 mg / kg to about 2.0 mg / kg, or It is about 1.0 mg / kg to about 2.0 mg / kg, or About 1.25 mg / kg to about 2.0 mg / kg, or About 1.5 mg / kg to about 2.0 mg / kg, or or about 0.5 mg / kg; or Approximately 0.75 mg / kg or; or or about 1 mg / kg; or It is about 1.5 mg / kg and; and / or BL8040 is: Administered via a pharmacologically acceptable route, or Administered subcutaneously, or Administered intravenously; and / or The last dose of BL8040 prior to the start of apheresis and / or leukocyte apheresis is: When apheresis and / or leukocyte apheresis treatment begins Administered about 120 hours to about 10 hours prior, or Administered about 72 hours to about 10 hours prior, or Administered about 24 hours to about 10 hours prior, or Administered about 14 to 10 hours prior, or Administered about 13 to 10 hours prior, or Administered about 13 to 10 hours prior, or Administered about 12 to 10 hours prior, or Administered about 11 to 10 hours prior, or Administered about 12 hours prior, or Administered about 11 hours prior, or Administered approximately 10 hours prior; and / or BL8040 is administered once daily; and / or BL8040 is administered 1, 2, 3, 4, or more than 4 times; and / or BL8040 is administered 5 times or less, 4 times or less, or 3 times or less, 2 times or less, or 1 time or less; and / or BL8040 is administered subcutaneously at about 1.5 mg / kg once daily for 1 day or up to 5 days, preferably the last dose of BL8040 prior to apheresis and / or leukocyte apheresis is administered at least about 10 hours before the apheresis and / or leukocyte apheresis treatment begins, preferably about 12 to about 10 hours before the apheresis and / or leukocyte apheresis treatment begins; and / or Here: The effective dose of a beta-adrenergic receptor inhibitor is: It is a pharmacologically effective dose with clinically acceptable serious side effects, or About 10 mg to about 480 mg per day, or About 20 mg to about 480 mg per day, or About 30 mg to about 480 mg per day, or About 40 mg to about 480 mg per day, or About 50 mg to about 480 mg per day, or About 60 mg to about 480 mg per day, or About 70 mg to about 480 mg per day, or About 80 mg to about 480 mg per day, or About 120 mg to about 480 mg per day, or About 160 mg to about 480 mg per day, or About 200 mg to about 480 mg per day, or About 240 mg to about 480 mg per day, or About 280 mg to about 480 mg per day, or About 320 mg to about 480 mg per day, or About 360 mg to about 480 mg per day, or About 400 mg to about 480 mg per day, or About 440 mg to about 480 mg per day, or About 10 mg to about 480 mg per day, or About 10 mg to about 450 mg per day, or It is about 10 mg to about 420 mg per day, or About 10 mg to about 390 mg per day, or About 10 mg to about 360 mg per day, or About 10 mg to about 330 mg per day, or About 10 mg to about 300 mg per day, or About 10 mg to about 270 mg per day, or About 10 mg to about 240 mg per day, or About 10 mg to about 210 mg per day, or It is about 10 mg to about 180 mg per day, or About 10 mg to about 150 mg per day, or It is about 10 mg to about 120 mg per day, or It is about 10 mg to about 90 mg per day, or It is about 10 mg to about 60 mg per day, or It is about 10 mg to about 30 mg per day, or It is about 10 mg to about 90 mg per day, or It is about 10 mg to about 60 mg per day, or It is about 10 mg to about 30 mg per day, or About 20 mg to about 480 mg per day, or About 20 mg to about 450 mg per day, or About 20 mg to about 440 mg per day, or About 20 mg to about 410 mg per day, or About 20 mg to about 400 mg per day, or About 20 mg to about 370 mg per day, or About 20 mg to about 360 mg per day, or About 20 mg to about 330 mg per day, or About 20 mg to about 320 mg per day, or About 20 mg to about 290 mg per day, or About 20 mg to about 280 mg per day, or About 20 mg to about 250 mg per day, or About 20 mg to about 240 mg per day, or About 20 mg to about 210 mg per day, or About 20 mg to about 200 mg per day, or About 20 mg to about 170 mg per day, or About 20 mg to about 160 mg per day, or About 20 mg to about 130 mg per day, or About 20 mg to about 120 mg per day, or It is about 20 mg to about 90 mg per day, or It is about 20 mg to about 80 mg per day, or It is about 20 mg to about 50 mg per day, or It is about 20 mg to about 40 mg per day, or About 20 mg to about 160 mg per day, or About 20 mg to about 130 mg per day, or About 20 mg to about 120 mg per day, or About 20 mg to about 100 mg per day, or It is about 20 mg to about 80 mg per day, or It is about 20 mg to about 70 mg per day, or It is about 20 mg to about 40 mg per day, or About 20 mg per day, or About 40 mg per day, or Approximately 60 mg per day, or Approximately 80 mg per day, and / or Beta-adrenergic receptor inhibitors are: Administered via a pharmacologically acceptable route, or Administered orally; and / or Beta-adrenergic receptor inhibitors are administered once daily, or twice daily, or three times daily, or four times daily; and / or Beta-adrenergic receptor inhibitors are: Administered at intervals of approximately 7 to 12 hours, or Administered at intervals of approximately 12 hours, or Administered at intervals of approximately 11 hours, or Administered at intervals of approximately 10 hours, or Administered at intervals of approximately 9 hours, or Administered at intervals of approximately 8 hours, or Administered at intervals of approximately 7 hours, or Administered at intervals of approximately 6 hours, or Administered at intervals of approximately 5 hours, or Administered at intervals of approximately 4 hours, or Administered at intervals of approximately 3 hours; and / or A beta-adrenergic receptor inhibitor is administered orally at a dose of about 30 mg, twice daily at intervals of about 7 to about 12 hours; and / or Here: The effective dose of an ADRB2 inhibitor is: It is a pharmacologically effective dose with clinically acceptable serious side effects, or About 10 mg to about 480 mg per day, or About 20 mg to about 480 mg per day, or About 30 mg to about 480 mg per day, or About 40 mg to about 480 mg per day, or About 50 mg to about 480 mg per day, or About 60 mg to about 480 mg per day, or About 70 mg to about 480 mg per day, or About 80 mg to about 480 mg per day, or About 120 mg to about 480 mg per day, or About 160 mg to about 480 mg per day, or About 200 mg to about 480 mg per day, or About 240 mg to about 480 mg per day, or About 280 mg to about 480 mg per day, or About 320 mg to about 480 mg per day, or About 360 mg to about 480 mg per day, or About 400 mg to about 480 mg per day, or About 440 mg to about 480 mg per day, or About 10 mg to about 480 mg per day, or About 10 mg to about 450 mg per day, or It is about 10 mg to about 420 mg per day, or About 10 mg to about 390 mg per day, or About 10 mg to about 360 mg per day, or About 10 mg to about 330 mg per day, or About 10 mg to about 300 mg per day, or About 10 mg to about 270 mg per day, or About 10 mg to about 240 mg per day, or About 10 mg to about 210 mg per day, or It is about 10 mg to about 180 mg per day, or About 10 mg to about 150 mg per day, or It is about 10 mg to about 120 mg per day, or It is about 10 mg to about 90 mg per day, or It is about 10 mg to about 60 mg per day, or It is about 10 mg to about 30 mg per day, or It is about 10 mg to about 90 mg per day, or It is about 10 mg to about 60 mg per day, or It is about 10 mg to about 30 mg per day, or About 20 mg to about 480 mg per day, or About 20 mg to about 450 mg per day, or About 20 mg to about 440 mg per day, or About 20 mg to about 410 mg per day, or About 20 mg to about 400 mg per day, or About 20 mg to about 370 mg per day, or About 20 mg to about 360 mg per day, or About 20 mg to about 330 mg per day, or About 20 mg to about 320 mg per day, or About 20 mg to about 290 mg per day, or About 20 mg to about 280 mg per day, or About 20 mg to about 250 mg per day, or About 20 mg to about 240 mg per day, or About 20 mg to about 210 mg per day, or About 20 mg to about 200 mg per day, or About 20 mg to about 170 mg per day, or About 20 mg to about 160 mg per day, or About 20 mg to about 130 mg per day, or About 20 mg to about 120 mg per day, or It is about 20 mg to about 90 mg per day, or It is about 20 mg to about 80 mg per day, or It is about 20 mg to about 50 mg per day, or It is about 20 mg to about 40 mg per day, or About 20 mg to about 160 mg per day, or About 20 mg to about 130 mg per day, or About 20 mg to about 120 mg per day, or About 20 mg to about 100 mg per day, or It is about 20 mg to about 80 mg per day, or It is about 20 mg to about 70 mg per day, or It is about 20 mg to about 40 mg per day, or About 20 mg per day, or About 40 mg per day, or Approximately 60 mg per day, or Approximately 80 mg per day, and / or ADRB2 inhibitors are: Administered via a pharmacologically acceptable route, or Administered orally; and / or ADRB2 inhibitors are administered once daily, or twice daily, or three times daily, or four times daily; and / or ADRB2 inhibitors are: It is administered at intervals of approximately 7 to 12 hours, or Administered at intervals of approximately 12 hours, or Administered at intervals of approximately 11 hours, or Administered at intervals of approximately 10 hours, or Administered at intervals of approximately 9 hours, or Administered at intervals of approximately 8 hours, or Administered at intervals of approximately 7 hours, or Administered at intervals of approximately 6 hours, or Administered at intervals of approximately 5 hours, or Administered at intervals of approximately 4 hours, or Administered at intervals of approximately 3 hours; and / or ADRB2 inhibitors are administered orally at a dose of about 30 mg, twice daily at intervals of about 7 to 12 hours; and / or Here: The effective amount of propranolol is: About 10 mg to about 480 mg per day, or About 20 mg to about 480 mg per day, or About 30 mg to about 480 mg per day, or About 40 mg to about 480 mg per day, or About 50 mg to about 480 mg per day, or About 60 mg to about 480 mg per day, or About 70 mg to about 480 mg per day, or About 80 mg to about 480 mg per day, or About 120 mg to about 480 mg per day, or About 160 mg to about 480 mg per day, or About 200 mg to about 480 mg per day, or About 240 mg to about 480 mg per day, or About 280 mg to about 480 mg per day, or About 320 mg to about 480 mg per day, or About 360 mg to about 480 mg per day, or About 400 mg to about 480 mg per day, or About 440 mg to about 480 mg per day, or About 10 mg to about 480 mg per day, or About 10 mg to about 450 mg per day, or It is about 10 mg to about 420 mg per day, or About 10 mg to about 390 mg per day, or About 10 mg to about 360 mg per day, or About 10 mg to about 330 mg per day, or About 10 mg to about 300 mg per day, or About 10 mg to about 270 mg per day, or About 10 mg to about 240 mg per day, or About 10 mg to about 210 mg per day, or It is about 10 mg to about 180 mg per day, or About 10 mg to about 150 mg per day, or It is about 10 mg to about 120 mg per day, or It is about 10 mg to about 90 mg per day, or It is about 10 mg to about 60 mg per day, or It is about 10 mg to about 30 mg per day, or It is about 10 mg to about 90 mg per day, or It is about 10 mg to about 60 mg per day, or It is about 10 mg to about 30 mg per day, or About 20 mg to about 480 mg per day, or About 20 mg to about 450 mg per day, or About 20 mg to about 440 mg per day, or About 20 mg to about 410 mg per day, or About 20 mg to about 400 mg per day, or About 20 mg to about 370 mg per day, or About 20 mg to about 360 mg per day, or About 20 mg to about 330 mg per day, or About 20 mg to about 320 mg per day, or About 20 mg to about 290 mg per day, or About 20 mg to about 280 mg per day, or About 20 mg to about 250 mg per day, or About 20 mg to about 240 mg per day, or About 20 mg to about 210 mg per day, or About 20 mg to about 200 mg per day, or About 20 mg to about 170 mg per day, or About 20 mg to about 160 mg per day, or About 20 mg to about 130 mg per day, or About 20 mg to about 120 mg per day, or It is about 20 mg to about 90 mg per day, or It is about 20 mg to about 80 mg per day, or It is about 20 mg to about 50 mg per day, or It is about 20 mg to about 40 mg per day, or About 20 mg to about 160 mg per day, or About 20 mg to about 130 mg per day, or About 20 mg to about 120 mg per day, or About 20 mg to about 100 mg per day, or It is about 20 mg to about 80 mg per day, or It is about 20 mg to about 70 mg per day, or It is about 20 mg to about 40 mg per day, or About 20 mg per day, or About 40 mg per day, or Approximately 60 mg per day, or Approximately 80 mg per day, and / or Propranolol is: Administered via a pharmacologically acceptable route, or Administered orally; and / or Propranolol is administered once daily, or twice daily, or three times daily, or four times daily; and / or Propranolol is: It is administered at intervals of approximately 7 to 12 hours, or Administered at intervals of approximately 12 hours, or Administered at intervals of approximately 11 hours, or Administered at intervals of approximately 10 hours, or Administered at intervals of approximately 9 hours, or Administered at intervals of approximately 8 hours, or Administered at intervals of approximately 7 hours, or Administered at intervals of approximately 6 hours, or Administered at intervals of approximately 5 hours, or Administered at intervals of approximately 4 hours, or Administered at intervals of approximately 3 hours; and / or Propranolol is administered orally at a dose of about 30 mg, twice daily at intervals of about 7 to about 12 hours; and / or Here: The effective amount of G-CSF is: Administered intravenously at about 1 µg / kg to about 70 µg / kg twice daily, or Administered by subcutaneous injection at a dose of about 1 µg / kg to about 40 µg / kg per day, or About 3 µg / kg to about 40 µg / kg per day by continuous subcutaneous injection, or About 10 ug / kg to about 40 ug / kg per day, or Approximately 10 ug / kg per day; and / or G-CSF is: Administered via a pharmacologically acceptable route, or Administered intravenously, or Administered subcutaneously, or Administered continuously subcutaneously; and / or The last dose of G-CSF inhibitor prior to the start of apheresis and / or leukocyte apheresis is: When apheresis and / or leukocyte apheresis treatment begins Administered about 120 hours to about 10 hours prior, or Administered about 72 hours to about 10 hours prior, or Administered about 24 hours to about 10 hours prior, or Administered about 14 to 10 hours prior, or Administered about 13 to 10 hours prior, or Administered about 13 to 10 hours prior, or Administered about 12 to 10 hours prior, or Administered about 11 to 10 hours prior, or Administered about 12 hours prior, or Administered about 11 hours prior, or Administered approximately 10 hours prior; and / or G-CSF is administered once daily; and / or G-CSF is administered 1, 2, 3, 4, 5, 6, 7, 8, or more than 8 times; and / or G-CSF is administered 8 times or less, 7 times or less, 6 times or less, 5 times or less, 4 times or less, or 3 times or less, 2 times or less, or 1 time or less; and / or A method in which G-CSF is administered by intravenous injection, subcutaneous injection, or serial subcutaneous injection at a rate of about 10 µg / kg per day for about 4 days, preferably at least about 10 hours prior to apheresis and / or leukocyte apheresis. Claim 116 In any one of paragraphs 98 through 115, CD34 in the subject + A drug that interferes with cell mobilization is administered, The drug is lenalidomide, and Lenalidomide is: Administered according to the drug label, and / or Administered at a dose of about 8 mg / kg to about 16 mg / kg, preferably about 16 mg / kg, and / or Administered via a pharmacologically acceptable route, or Administered intravenously, or Administered for about 3 to about 4 cycles; and / or The drug is daratumumab, and Daratumumab is: Administered according to the drug label, and / or Administered at a dose of about 8 mg / kg to about 16 mg / kg, preferably about 16 mg / kg, and / or Administered via a pharmacologically acceptable route, or Administered intravenously, or A method administered for about 3 to 4 cycles. Claim 117 In any one of paragraphs 98 through 116, The method is: Does not cause adverse events in the subject during or after treatment, or Does not cause severe adverse reactions in subjects during or after treatment, or Does not cause adverse reactions of grade 1 or 2 or greater severity, or A method in which adverse reactions, if caused, are limited to a list consisting of bone pain, hypocalcemia, diarrhea, dry mouth, nausea, hypokalemia, peripheral edema, paranesthesia, etc.

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