MX2008002374A - Methods to enhancechemotherapy - Google Patents

Abstract

Antagonists of the interaction of CXCR4 receptor with its ligand enhance the effectiveness of chemotherapeutic methods in subjects afflicted with myeloid or hematopoietic malignancies.

Description

METHODS TO IMPROVE CHEMOTHERAPY RELATED REQUESTS This application claims the benefit of the provisional applications of E.U.A. serial numbers 60 / 709,978, filed on August 19, 2005, and 60 / 734,736, filed on November 8, 2005. The contents of these documents are hereby incorporated herein by reference in their entirety.

TECHNICAL FIELD The invention is in the field of the treatment of cancers related to hematopoietic cells. More particularly, the invention relates to methods for improving the chemotherapy of said conditions.

TECHNICAL BACKGROUND A common procedure for hematopoietic cell-related cancers, such as myelolden leukemias and lymphoid leukemias, is a chemotherapy session to destroy the malignant cells combined with the transplantation of hematopoietic progenitor cells of either autogenic or allogeneic origin. It is believed that the lack of success frequently experienced with this treatment regimen is due to the failure of chemotherapy to completely eliminate the malignant hematopoietic cells or their precursors. The present invention improves this method by coupling it with the administration of a compound that improves the effect of chemotherapy with respect to these residual premalignant or malignant cells. The compounds useful in the method of the invention are antagonists of the CXCR4 receptor that prevent their interaction with the cytokine factor 1 derived from stromal cells (SDF-1). Many such agents are known in the art. Such agents are described, for example, in the patents of E.U.A. Nos. 5,021, 409; 6,001, 826; 5,583,131; 5,698,546; 5,817,807 and 6,506,770, incorporated herein by reference, and in PCT publications WO 92/16494; WO 93/12096; WO 95/18808; WO 00/02870; WO 00/56729; WO 01/44229; WO 02/22600; WO 02/22599; WO 02/34745, WO 03/055876; WO 04/091518 and WO 04/093217, also incorporated herein by reference. The present inventors have previously found, and have described in PCT publication WO 02/58653, that certain antagonists of CXCR4, in particular, AMD3100, have the effect of increasing the white blood cell count. They have also found, and have described in PCT publication WO 03/011277, that these antagonists have the effect of mobilizing progenitor cells and / or stem cells from the bone marrow into the circulating blood.

The chemokine receptor CXCR4 and its natural ligand SDF-1 appear to be important in the process of hematopoiesis (for reviews, see Maekawa, T., et al., Interna! Med. (2000) 39: 90-100; Nagasawa, T., et al., Int. J. Hematol. (2000) 72: 408-411). For example, mice with blocked expression in CXCR4 or SDF-1 exhibit hematopoietic defects (Ma, Q., et al., Proc. Nati, Acad. Sci USA (1998) 95: 9448-9453). It seems that SDF-1 is able to control the positioning and differentiation of cells that have CXCR4 receptors, whether these cells are stem cells (ie cells that are CD34 +) or progenitor cells (which result in the formation of specified types of colonies in response to particular stimuli). It appears that, within the bone marrow microenvironment, SDF-1 acts as a potent chemoattractant of mature and immature hematopoietic cells, and in this way the expression of CXCR4 on leukemic progenitor cells and leukemia cells can contribute by guiding them towards the microenvironment of the bone marrow Elevated levels of CXCR4 are detected in leukemic cells of patients with chronic B-cell lymphocytic leukemia (B-CLL) (Mohle, R., et al., Leukemia (1999) 13: 1954-1959). It also appears to be that autocropic secretion of SDF-1 by adherent cell-like cells derived from blood in chronic lymphocytic leukemia (CLL) protects leukemic B cells from spontaneous apoptosis (Burger, JA, et al., Blood (2000 ) 96: 2655-2663). No increased levels were detected in leukemic cells from patients with T-ALL or leukemic cells from patients with AML according to Mohle, et al., Cited above.; Voermans, C, et al., Leukemia (2002) 16: 650-657; Bradstock, K. F., et al., Leukemia (2000) 14: 882-888; Dialynas, D.P., et al., Stem Cells (2001) 19: 443-452; Shen, W., et al., Exp. Hematol. (2001) 29: 1439-1447. However, it appears that CXCR4 expression levels vary among various types of AML, as reported by Rombouts, E. J., et al., Blood (2004) 104: 550-557; Fukuda, S., et al., Blood (2005) 105: 3117-3126. It is also reported that CXCR4 mediates the arrival and grafting of pre-B-ALL and AML cells to the bone marrow, although other factors may be involved (Shen, et al., Cited above, Tavor, S., et al., Cancer Res. (2004) 64: 2817-2824). It was recently shown, in an in vitro context, that AMD3100 blocked the qulmiotaxis induced by SDF-1 of pre-B-ALL cells in stromal layers of bone marrow, and increased the cytotoxic and antiproliferative effects of vincristine and dexamethasone (Juarez, J ., et al., Leukemia (2003) 17: 1294-1300). These studies suggest that interactions between SDF-1 and CXCR4 are involved in the microenvironmental regulation of leukemic cells, and that such interaction plays a role in the resistance of residual post-chemotherapy AML exposure to additional chemotherapeutic agents. There is a need to mobilize precancerous or cancerous cells of the bone marrow and in the peripheral blood system, where these cells can be exposed to chemotherapeutic agents. The present invention addresses this need by the use of inhibitors of the CXCR4 receptor that enhance the effects of standard chemotherapeutic agents, through the release and / or rapid movement of pre-leukemic cells and leukemic cells of the bone marrow microenvironment and in circulating blood before, or during or after, treatment by chemotherapy. This invention can be used to treat subjects who may or may not require transplantation. It is not intended that the citation of the foregoing documents be an admission that any of the foregoing is pertinent prior art. All statements regarding the date or representation as to the content of these documents are based on information available to applicants, and do not constitute any admission as to the accuracy of the dates or content of these documents, and does not purport to that are limited by some theory or hypothesis described in these documents. In addition, all documents referred to throughout this application are hereby incorporated by reference in their entirety.

BRIEF DESCRIPTION OF THE INVENTION The invention is directed to methods of treating animal subjects, in particular, veterinary and human subjects, with chemotherapeutic methods, while also administering a CXCR4 antagonist that improves the effect of chemotherapy.

Thus, in one aspect, the invention is directed to a method for treating a subject having a hematopoietic malignancy, such as a lymphoma, a myeloma or a leukemia, which method comprises administering one or more CXCR4 antagonists and one or more chemotherapeutic agents. The CXCR4 antagonists can be administered before, during and / or after the chemotherapeutic regimens are administered. In another aspect, the invention is directed to pharmaceutical or veterinary compositions comprising a CXCR4 antagonist for use in the method of the invention. These compositions comprise one or more CXCR4 antagonists together with suitable pharmaceutically or veterinarily acceptable excipients. Some of the antagonists useful in the invention are those described in the patents of E.U.A. Nos. 5,021, 409; 6,001, 826; 5,583,131; 5,698,546; 5,817,807 and 6,506,770, incorporated herein by reference, and in PCT publications WO 92/16494; WO 93/12096; WO 95/18808; WO 00/02870; WO 00/56729; WO 01/44229; WO 02/22600; WO 02/22599; WO 02/34745, WO 03/055876; WO 04/091518 and WO 04/093817, also incorporated herein by reference. Peptide-based antagonists are described in WO 2001/85196; WO 2000/09152 and WO 99/47158. The use of antibodies as inhibitors of CXCR4 that interact with their ligand is described in WO 99/50461. Other compounds include T22 (Murakami, T., et al., J. Exp. Med., 186: 1389-1393 (1997)), ALX40-4C (Doranz, BJ, et al., J. Exp. Med., 186, 1395-1400 (1997)); Donzella, G. A., Nat. Med., 4, 72-77 (1998)), and the like. As for the methods of preparation of these substances, they can be found, for example, in J. Exp. Med., 186, 1189-1191 (1997), with some conventional modifications.

DETAILED DESCRIPTION OF THE INVENTION The invention uses compounds that inhibit the binding of SDF-1 to CXCR4 (CXCR4 antagonists). While not wishing to be bound by any theory, compounds that inhibit the binding of SDF-1 to CXCR4 effect the improvement of chemotherapy by virtue of such inhibition, depriving malignant or premalignant cells of the protection of stromal cells. of the bone marrow. As used herein, the term "premalignant cells" refers to cells that can form hematopoietic or malignant myeloid cells. The hematopoietic or malignant myeloid cells are those that characterize the conditions of myeloma, leukemia and lymphoma. Particular forms of these diseases include acute myelitis leukemia (AML), acute lymphatic leukemia (ALL), multiple myeloma (MM), chronic myelogenous leukemia (CML), hairy cell leukemia (HCL), acute promyelocytic leukemia (APL), lymnocytic leukemia chronic (CLL), and several types. Chemotherapeutic compounds, or agents that can be used in the methods whose effectiveness is improved by the methods of the invention, include carboplatin, carmustine, chlorambucil, dacarbazine, ifosfamide, lomustine, mechlorethamine, procarbazine, pentostatin, (2'-deoxyco-funicin), etoposide, teniposide, topotecan, vinblastine, vincristine, paclitaxel, dexamethasone, methylprednisolone, prednisone, all-trans retinoic acid, arsenic trioxide, interferon-alpha, rituximab (Rituxan®), gemtuzumab, ozogamicin, imatinib mesylate, cytarabine (cytosine arabinoside, Ara -C, Cytosar-U), melphalan, busulfan (Myleran®), thiotepa, bleomycin, platinum (cisplatin), cyclophosphamide, Cytoxan®, daunorubicin, doxorubicin, darrubicin, mitoxantrone, 5-azacytidine, cladribine, fludarabine, hydroxyurea, -mercaptopurine, methotrexate, 6-tloguanin, and many others. A wide variety of chemotherapeutic methods are available in the art. The invention now uses these standard methods or variations thereof but, in addition, it provides the administration of CXCR4 antagonists to improve the effect of said methods. Preferably, these antagonists are administered prior to and / or concomitant with, the patient's exposure to said methods. Administration may continue after the method has also stopped, if desired. The dosage levels and the administration mode are interdependent. When administered subcutaneously, for example, the dosage levels are in the range of 50 μg / kg - 1 mg / kg, preferably 200 μg / kg - 500 μg / kg. Dosage levels using oral administration may be higher, and intravenous administration somewhat less.

In some embodiments, the CXCR4 antagonist is of the formula: Z-linker-Z '(1) wherein Z is a cyclic polyamine containing 9-32 ring members of which 2-8 are nitrogen atoms, said atoms of nitrogen being separated from each other by at least 2 carbon atoms, and wherein said heterocycle may optionally contain additional heteroatoms in addition to nitrogen, and / or may be fused to an additional ring system; or Z is of the formula: wherein A comprises a monocyclic or bicyclic fused ring system containing at least one N, and B is H or an organic portion of 1-20 atoms; Z 'can be described in a form as defined by the above Z, or it can alternatively be of the formula: -N (R) - (CR2) "-X wherein each R is independently H or straight, branched or cyclic (1-6 carbons), n is 1 or 2, and X is an aromatic ring, including heteroaromatic rings, or is a mercaptan; or Z 'may be absent and the compound of formula 1 ends with the portion defined below as a linker; "linker" represents a bond, alkylene (1-6 carbons), or may comprise aryl, fused aryl, oxygen atoms contained in an alkylene chain, or may contain keto groups or nitrogen or sulfur atoms. Specific forms of the compounds of formula (1) are discussed below. In the compounds of formula (1), some embodiments of Z and Z 'are cyclic polyamine portions having 9-24 carbons including 3-5 nitrogen atoms, eg, 1, 5,9,13-tetraazacyclohexadecane; 1, 5,8,11, 14-pentaazacyclohexadecane; 1, 4,8,11-tetraazacyclotetradecane; 1, 5,9-triazacyclododecane; 1, 4,7,10-tetraazaclclododecane; and the like, including said cyclic polyamines which are fused to an additional aromatic or heteroaromatic ring and / or which contain a heteroatom minus nitrogen incorporated in the ring. Modalities wherein the cyclic polyamine contains a further fused cyclic system or one or more additional heteroatoms, are described in the U.S. patent. No. 5,698,546 and WO 01/44229, incorporated herein by reference. Other embodiments are 3,7,11, 17-tetraazabiclclo (13.3.1) heptadeca-1 (17), 13,15-triene; 4,7,10,17- tetraazabicyclo (13.3.1) heptadeca-1 (17), 13,15-triene; 1, 4,7,10-tetraazacyclotetradecane; 1, 4,7-triazacyclotetradecane; and 4,7,10-triazabicyclo (13.3.1) heptadeca-1 (17), 13, 15-triene. Some embodiments of the compound of the formula (1) include 2,2'-bicyclamate; 6,6'-biciclama; and the modalities set out in the patents of E.U.A. Nos. 5,021, 409 and 6,001, 826, and in particular 1, 1 '- [1,4-phenylene-bis (methylene)] - bis-1,4,8,11-tetraazacyclotetradecane, disclosed in the patent of E.U.A. No. 5,583,131, and designated herein as AMD3100. In some embodiments, Z is 1, 4,8,11-tetraazacyclotetradecane, the linker is 1, 3- or 1,4-phenylene-bis (alkylene), in particular 1,4-phenylene-bjs (methylene), and Z 'is -NR (CR2) nX, where X is pyridine, and in particular where Z' is NHCH2CH2-pyridine. In some embodiments, the compound is AMD3465, which is N- [1,4,8,11-tetraazacyclotetradecanyl- (1,4-phenylene-bis- (methylene)] - 2-aminoalkylpyridine, or substituted forms thereof. When Z 'is less a cyclic polyamine as defined in Z, some embodiments are set forth in U.S. Patent Nos. 5,817,807 and 6,506,770, also incorporated herein by reference.Some forms wherein Z is of the formula: A / B wherein A comprises a monocyclic or bicyclic fused ring system containing at least one N, and B is H or an organic portion of 1-20 atoms, are described in WO 00/56729; WO 02/22600; WO 02/34745; WO 02/22599 and WO 03/55876 cited above, and all incorporated herein by reference. In one embodiment, as set forth in WO 03/55876, A is 5,6,7,8-tetrahydroquinolin-8-ylo, and B is 1 H-benzimidazol-2-yl-methyl. In some of these modalities, Z 'is absent and the linker is an omega aminoalkyl group. In this manner, an illustrative compound is AMD11070, which is N1- (1 H-benzimidazol-2-ylmethyl) -N1- (5,6,7,8-tetrahydroquinolin-8-yl) butane-1, 4- diamine. In this way, important modalities include AMD11070, and substituted forms thereof. Ways of the linker portion include those in which the linker is a bond, or wherein the linker includes an aromatic moiety flanked by alkylene, preferably methylene moieties. The linking groups include the grouped methylene forms of 1,3-phenylene, 2,6-pyridine, 3,5-pyridine, 2,5-thiophene, 4,4 '- (2,2, -bipyrimidine); 2.9- (1, 10-phenanthroline), and the like. A particularly preferred linker is 1,4-phenylene-bis- (methylene). Additional compounds that are antagonists of CXCR4 are described in the applications of E.U.A. 10 / 823,494, filed on April 12, 2004, and 10/831, 098, filed on April 22, 2004, and 11 / 012,002, filed on December 13, 2004, incorporated herein by reference.

CXCR4 inhibitors that can be used to implement the methods of the invention also include, but are not limited to, CTCE-0214; CTCE-9908; CP-1221 (linear peptides, cyclic peptides, natural amino acids, non-natural amino acids and peptidomimetic compounds); T140 and analogs thereof; 4F-benzoyl-TN24003; KRH-1120; KRH-1636; KRH-2731; polyphemusin analogues; ALX40-4C; or those described in WO 01/85196; WO 99/50461; WO 01/94420; and WO 03/090512, each of which is incorporated herein by reference. Methods for synthesizing the compounds useful in the method of the invention are set forth in the patents and applications of E.U.A. above, as well as in the patent of E.U.A. 6,489,472 and the application of E.U.A. 11 / 077,896, filed March 11, 2005, incorporated herein by reference. Additional CXCR4 inhibitors are set forth in Appendix A. As indicated above, AMD3100 is an example of an antagonist with the chemokine receptor CXCR4 (Gerlach, et al., J. Biol. Chem. (2001) 276: 14153-14160). . This compound interferes with the binding of SDF-1 derived from bone marrow stromal cells to CXCR4 in stem cells, which leads to the release of hematopoietic stem cells from the bone marrow into the circulation (Broxmeyer, et al., Blood (2001 ) 98: 811a (summary)). The compounds of the invention can be prepared in the form of prodrugs, ie, protected forms that release the compounds of the invention after their administration to the subject. Typically, the protecting groups are hydrolyzed in body fluids such as in the blood stream, thereby releasing the active compound, or are oxidized or reduced in vivo to release the active compound. A discussion of prodrugs is found in Smith and Williams, Introduction to the Principles of Druq Desiqn, Smith, H. J .; Wright, 2a. ed., London (1988). Compounds useful in the invention which are amines, may be administered or prepared in the forms of their acid addition salts or metal complexes thereof. Suitable acid addition salts include salts of inorganic acids that are biocompatible, including HCl, HBr, sulfuric acid, phosphoric acid, and the like, as well as organic acids such as acetic acid, propionic acid, butyric acid, and the like, as well as acids that they contain more than one carboxyl group, such as oxalic acid, glutaric acid, adipic acid, and the like. Typically, at physiological pH, the compounds of the invention will be in the forms of the acid addition salts. Compounds useful in the invention which are carboxylic or otherwise acidic acids, may be administered or prepared in salt forms formed from organic or inorganic bases which are physiologically compatible. In this manner, these compounds can be prepared in the forms of their sodium, potassium, calcium or magnesium salts, as appropriate, or they can be salts with organic bases such as caffeine or ethylamine. These compounds can also be in the form of metal complexes. When prepared as purified forms, the compounds can also crystallize like hydrates or other solvates. Those forms of the compounds used in the invention that contain chiral centers may be optically pure, or may contain a mixture of stereoisomers, including racemic mixtures or mixtures of varying optical purity. CXCR4 antagonists can be formulated for administration to an animal subject, using commonly understood formulation techniques well known in the art. Formulations that are suitable for particular modes of administration and for compounds useful in the invention can be found in Remington's Pharmaceutical Sciences, latest edition, Mack Pubiishing Company, Easton, PA. Preferably, CXCR4 antagonists are administered by injection, more preferably by intravenous injection, but also by subcutaneous or intraperitoneal injection, and the like. Additional parenteral routes of administration include intramuscular and intra-articular injection. For intravenous or parenteral administration, the compounds are formulated in suitable liquid form with excipients, as required. The compositions may contain liposomes or other suitable vehicles. For intravenous injection, the solution is made isotonic using standard preparations such as Hank's solution.

In addition to administration by injection, other routes of administration may also be used. The compounds can be formulated into tablets, capsules, syrups, powders, or other forms suitable for oral administration. By the use of suitable excipients, these compounds can also be administered through the mucosa using suppositories or intranasal sprays. Transdermal administration can also be effected using suitable penetrants, and controlling the rate of release. The chosen formulation and route of administration will be adapted to the individual subject, the nature of the condition to be treated in the subject, and in general, the judgment of the specialist in charge. Suitable dosing scales for CXCR4 antagonists vary according to these considerations but, in general, the compounds are administered on the scale of about 0.1 μg / kg-5 mg / kg body weight; preferably, the scale is from about 1 μg / kg-500 μg / kg to 1 mg / kg of body weight. For a typical human subject of 70 kg, in this way, the dosing scale is approximately 0.7 μg / kg-350 mg. The dosages may be higher, when the compounds are administered orally or transdermally in comparison, for example, with intravenous administration. The CXCR4 antagonists can be administered as an individual bolus dose, or doses over time, as in intravenous or transdermal administration, or in multiple dosages. CXCR4 antagonists can be administered together with other factors that facilitate mobilization, or other factors that are nutritional or therapeutically beneficial. Additional factors can be administered in the same composition, in different compositions, but simultaneously, or in a tandem protocol, with the administration of the CXCR4 antagonist. Additional factors that may be included are recombinant G-CSF (Neupogen®, Granocyte® / Neutrogin® and Stemgen®), a covalent conjugate of recombinant G-CSF (Neulasta®), granuloclose-macrophage colony stimulating factor (GM-). CSF) (such as Leukine® and Leucomax®), interleukin-1 (IL-1), interleukin-3 (IL-3), interleukin-8 (IL-8), PIXY-321 (fusion protein GM-CSF / IL-3), inflammatory protein of macrophages, stem cell factor and thrombopoietin, as well as antibiotics, vitamins, herbal extracts, anti-inflammatory, nutrients, antipyretics, analgesics, cyclophosphamide, and the like. As indicated above, the compounds are administered in conjunction with chemotherapeutic methods. These methods are those generally used in the treatment of hematopoietic or myelitic malignancies that are subjected to treatment by the method of the invention. A wide variety of such methods is known in the art. Generally, subjects who will respond favorably to the method of the invention include medical and veterinary subjects, including human patients. Among other subjects for whom the methods of the invention are useful, are cats, dogs, large animals, birds such as chickens, and the like, less standard research rodents such as mice, rabbits or laboratory rats. In general, any subject exhibiting a hematopoietic or myelitic malignancy would benefit from the methods of the invention. A wide variety of chemotherapeutic protocols are used, many of said protocols including combinations of drugs administered simultaneously or in tandem. CXCR4 antagonists can be administered at several points in the simultaneous or tandem protocols. For example, a protocol for AML involves combinations of busulfan and fludarabine. These drugs are administered intravenously. The CXCR4 antagonist can be administered several hours before the first administration of this drug, which is repeated for several days. The CXCR4 antagonist can be administered every day before or during the administration of fludarabine, or only busulfan is typically administered subsequent to fludarabine for several days, and the CXCR4 antagonist may be administered each day together with, before, or after administration of busulfan, administration before, during or after treatment may be required. . Various combinations of the above agents are used in such protocols, and the regulation and frequency of the administration of CXCR4 is subject to routine optimization, well known to those skilled in the art.

Having now generally described the invention, it will be more readily understood by reference to the following examples, which are provided by way of illustration, and are not intended to be limiting of the present invention, unless otherwise specified.

EXAMPLE 1 This example describes a murine model of human acute promyelocytic leukemia (APL) used to determine the effect of a CXCR4 antagonist on the mobilization of APL cells in the peripheral blood, and on the sensitivity to chemotherapeutic agents that are known to affect proliferation. of these cells. Murine APL cells were generated by "replacing the expression" of PML-RARa cDNA from human APL cells at the murine cathepsin G locus (Westerveit, et al., PubMed (2003) 102 (5): 1857-1865) that resulted in overexpression in the compartment of murine promyelocytes. Mouse APL cells, after injection into syngeneic receptors, are preferably directed towards the bone marrow microenvironment in a manner similar to that seen in human AML, and expand there for 20 to 30 days after circulating in high numbers in the peripheral blood. This eventually leads to the death of the animals around 50 to 80 days more.

By using a murine model of APL, it can be determined whether the leukemic cells are "mobilized" in a manner similar to normal stem cells, after treatment with a test compound, such as AMD3100, AMD3465, AMD11070 and other compounds described in the present. In one example, AMD3100 (5 mg / kg) injected immediately at the same time when APL cells were injected, had no impact on the graft (short or long term) of normal bone marrow stem cells or leukemic cells. However, where AMD3100 was administered 11 days after the APL injection, a rapid mobilization of the leukemic cells was observed. Forty percent (2/5) of the mice that received a single dose of AMD3100 on the day +11 after the injection of APL, died 2 to 4 hours after administration of AMD3100. It was observed that the administration of AMD3100 on day +11 induced a 3-fold increase in the total white blood cell count (WBC), and a 10-fold increase in the peripheral blood leukemic bla When AMD3100 was administered concomitantly with cytarabine (200 mg / kg) on day +11 in mice, this treatment significantly prolonged the overall survival of the mice, compared to mice treated with cytarabine alone. Based on the observed results, it may be possible that tumor resistance can be overcome by enhancing the effects of chemotherapeutic agents, by mobilizing tumor cells from the bone marrow in the peripheral blood.

EXAMPLE 2 Clinical study The in vivo effect of the CXCR4 antagonist AMD3100 was studied in three patients with AML, who had insufficient mobilization of CD34 + cells for autologous stem cell transplantation with G-CSF and / or Cytoxan®. The combination of G-CSF and AMD3100 (for 3-4 days) resulted in massive mobilization of leukemic cells in the circulation in a time-dependent manner, as determined by flow cytometry and FISH interphase analysis of their respective cytogenetic abnormalities.

EXAMPLE 3 In vitro data In a previously demonstrated study, it was shown that the interactions between stroma and leukemia mediate the protection of leukemic cells against apoptosis induced by chemotherapy (Konopleva, M., Leukemia (2002): 1713-1724). Co-culture systems of AML cells with in vitro stromal cells showed that the stromal cells significantly protected the leukemic cells (p <0.01). The application of AMD3465 decreased the stromal-mediated protection against apoptosis by AraC and busulfan, and down-regulated AKT signaling in AML cells.

EXAMPLE 4 Animal model In a murine model of Baf-FLT3ITD leukaemias labeled with luciferase, AMD3465 induced massive spread of leukemia, which was suppressed by treatment with sorafenib, a potent inhibitor of FLT3ITD (Zhang, ASH 2006).

Appendix A Examples of CXCR4 antagonists of formula 1 include compounds of formula (1A): V-CR2-Ar1-CR2 NR- (CR2)? - Ar2 (1A) wherein V is a substituted 9-24 member heterocycle containing 2-4 optionally substituted amine nitrogen atoms spaced apart from each other by 2 or more optionally substituted carbon atoms, and which heterocycle optionally may comprise a ring aromatic or heteroaromatic fused, and wherein: (a) said heterocycle contains at least one O or S, said O u S spaced from any adjacent heteroatom by at least 2 carbon atoms, and wherein said S is optionally oxidized, or (b) at least one carbon atom in said ring is replaced by an avid electron substituent, or (c) (a) and (b); and wherein each R is independently H or a straight, branched or cyclic chain alkyl containing 1-6 carbons; x is 0-4; Ar 1 is a substituted or unsubstituted aromatic or heteroaromatic moiety; and Ar2 is a substituted or unsubstituted aromatic or heterocyclic group. In another embodiment of formula 1, the CXCR4 antagonist has the formula: V-CH2-Ar1-CH2NR-CH2-Ar * wherein V is a heterocycle as defined in formula (1A), and wherein: (a) said heterocycle is substituted with halo or = 0; or (b) said heterocycle contains O or S; or (c) (a) and (b), and wherein Ar 1 is 1, 3 or 1,4-unsubstituted phenylene, R is H, methyl or ethyl, and Ar 2 is pyridinyl or unsubstituted phenyl. Preferred modes of x are 0-2 and 1-2. The heterocycle V can contain 3N and at least one carbon atom in the heterocycle which is substituted by at least one fluoro substituent. The portion R can independently be hydrogen or methyl. The number of groups (CR2) X can be 0-4, 0-2 or 1-2. The Ar1 portion can be 1, 3 or 1, 4-phenylene. The Ar2 portion can be phenyl or pyridyl. The heterocycle V can be a 12-16 member heterocycle, or it can contain O or S as a ring member. The heterocycle V may also contain an oxidized sulfur as a ring member. In one example, at least one carbon in the heterocycle V is replaced by = O. Compounds of formula (1A), and methods for synthesizing said compounds are described in WO 01/44229, incorporated herein by reference. Related to these compounds, are the compounds having the formula (1B): V-CR1R2-Ar-CR3R4-N (R5) - (CR6R7) X-R8 (1B) wherein V is a system of 1, 4 , 8, 11 -tetraazacyclotetra-decanyl, 4,7,10,17-tetraazabicyclo [13.3.1] heptadeca-1 (17), 13,15-trienyl, 1, 4,7-triazacyclotetra-decanyl, 4,7,10-triazabicyclo [13.3.1] heptadeca-1 (17), 13,15-trienyl, 1,7-diazacyclotetradecanyl or 4,10-diazabicyclo [13.31.1] heptadeca-1 (17), 13,15-trienyl; R1 to R7 may be the same or different, and are independently selected from hydrogen or straight, branched or cyclic C-? 6 alkyl; R8 is pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, thiophen-yl, thiophenyl, aminobenzyl, piperidinyl, purine, piperazinyl, phenylpiperazinyl or mercaptan; Ar is a phenylene ring optionally substituted at individual or multiple positions with alkyl, aryl, amino, alkoxy, hydroxy, halogen, carboxyl and / or carboxamido; and x is 1 or 2. In the formula (1B) above, the portion V can be optionally substituted by hydroxyl, alkoxy, thiol, thioalkyl, halogen, nitro, carboxy, amido, sulfonic acid and / or phosphate. Compounds of formula (1B), their pharmaceutically acceptable salts or metal complexes thereof, and methods for synthesizing said compounds are described in WO 00/02870, which is incorporated herein by reference. Other inhibitors of CXCR4 are of the formula (1C): wherein V2 is a system of 1, 4,8,11-tetraazacyclotetra-decanyl or 4,7,10,17-tetraazabicyclo [13.3.1] heptadeca-1 (17 ), Optionally substituted 13,15-trienyl; 9 and R10 may be the same or different, and are independently selected from hydrogen or straight, branched or cyclic C 1-6 alkyl; Ar2 is an aromatic or heterocyclic ring, each being optionally substituted in individual or multiple positions with electron donor or avid groups and / or aromatic and heterocyclic groups and alkyl derivatives thereof, and acid addition salts and metal complexes thereof. In the formula (1C) above, Ar 2 may be optionally substituted with alkyl, aryl, amino, alkoxy, hydroxy, halogen, carboxyl and / or carboxamido. In particular examples, Ar2 is optionally substituted with alkoxy, alkyl or halogen. Compounds having the formula (1C), and methods for synthesizing them, are described in WO 00/02870, incorporated herein by reference. Other CXCR4 antagonists are of the formula (1 D): VRA-R'-W (1 D) wherein V and W are independently cyclic polyamine portions having from 9 to 32 ring members and from 3 to 8 nitrogens of amine in the ring spaced by 2 or more carbon atoms from each other, and having one or more aromatic or heteroaromatic rings fused therewith, A is an aromatic or heteroaromatic portion when V and W have one or more aromatic or heteroaromatic portions fused with it, with or without an additional heteroatom less nitrogen incorporated in the ring, or A is an aromatic or heteroaromatic portion when V and W contain a heteroatom minus nitrogen incorporated in the ring without having one or more aromatic or heteroaromatic portions fused with the same, and R and R 'are each a substituted or unsubstituted alkylene chain or heteroatom containing chain that separates the cyclic polyamines and the A portion. the (1D) above, R and R 'may each be methylene. In one example, A is 1, 3- or 1, 4-phenylene. In another example, each V and W is a substituted or unsubstituted bicyclic or tricyclic ring system containing only carbon and nitrogen atoms in the rings. One of the cyclic ring systems may be a 10 to 20 member polyamine ring system having from 3 to 6 amine nitrogen atoms, and the ring system or systems are a fused benzyl or pyridinyl ring system . Compounds having the formula (1 D), and methods for synthesizing said compounds, are described in the U.S. patent. 5,698,546, incorporated herein by reference. Other CXCR4 antagonists are of the formula (1 E): Z- R- A- R'- Y (1 E) wherein Z and Y are identical cyclic polyamine portions having 10 to 15 ring members and 3 ring members. to 6 amine nitrogens in the ring spaced by 2 or more carbon atoms from each other, said amine nitrogens being the only ring heteroatoms, A is an aromatic or heteroaromatic portion less quinoline, R and R 'are each methylene linked to Nitrogen atoms in Z and Y, the amine nitrogen atoms being otherwise unsubstituted. In the formula (1 E) above, each Z and Y portion may have 14 ring members and 4 amine nitrogens in the ring. Compounds having the formula (1 E), and methods for synthesizing said compounds, are described in the U.S. patent. 5,583,131, incorporated herein by reference. The CXCR4 antagonist can be of the formula (1 F): Z- (A) nY (1 F) wherein Z and Y are independently cyclic polyamine portions having from 9 to 32 ring members, and from 3 to 8 amine nitrogen atoms in the ring, A is an atom or linking group, and n is O or an integer from 1 to 6. In the formula (1 F) above, each Z and Y portion may have from 10 to 24 members of ring, or 12 to 18 ring members. Each Z and Y portion can also have from 4 to 6 nitrogen atoms of amine in the ring. In one example, n is 0. In another example, A is methylene. Compounds having the formula (1 F), and methods for synthesizing said compounds, are described in the patent of E.U.A. 5,021, 409, incorporated herein by reference. Other CXCR4 antagonists are of the formula (2A): Y- W (CRW ^ rCR ^ NfRSMCRdR7) ^ 8 z (2A) wherein W is a nitrogen atom and Y is vacant, or W is a carbon atom and Y = H; R1 to R7 may be identical or different, and are independently hydrogen or straight, branched or cyclic C1-6alkyl; R8 is an optionally substituted heterocyclic group or an optionally substituted aromatic group; Ar is an aromatic or heteroaromatic ring optionally substituted at individual or multiple non-bonding positions, with electron donating or electron aviding groups; n and n 'are independently 0-2; X is a group of the formula: A B V wherein ring A is a 5 or 6 membered saturated or unsaturated ring, optionally substituted, and P is an optionally substituted nitrogen atom, and wherein any heteroatom in addition to P in ring A is N; wherein ring B is an optionally substituted 5 to 7 membered ring; wherein ring A or ring B is attached to group W of any position through group V; where V is a chemical bond, or V is a group (CH2) n "(where n" = 1-2), or V is a group C = 0; and wherein Z is selected from the group consisting of: a hydrogen atom; an optionally substituted C-? -6 alkyl group; an optionally substituted aromatic or heterocyclic group; an optionally substituted amino group; an alkylamino group of C -? - 6 or cycloalkylamino of C3-7 optionally substituted; or a substituted carbonyl group; or pharmaceutically acceptable acid addition salts thereof; wherein said compound may be in any stereoisomeric form, or may be present as a mixture of stereoisomeric forms thereof; wherein ring B is selected from the group consisting of: benzene and a 5 to 7 membered cycloalkyl ring; and optionally substituted forms thereof. CXCR4 antagonists also include compounds of formula (2B): X Y- W (CR1R2) nArCR3R4N (R5) (CR6R7) n.Rβ z (2B) wherein W is a nitrogen atom and Y is vacant; R1 to R7 may be the same or different, and are independently hydrogen or straight, branched or cyclic C-? -6 alkyl; R8 is an optionally substituted heterocyclic group or an optionally substituted aromatic group; Ar is an aromatic or heteroaromatic ring optionally substituted at individual or multiple non-linking positions, with electron donating or electron aviding groups; n and n 'are independently 0-2; X is a group of the formula: wherein ring A is a saturated or unsaturated 5 or 6 membered ring, optionally substituted, and P is an optionally substituted nitrogen atom, and wherein any heteroatom in ring A or B is N; wherein ring B is an optionally substituted 5 to 7 membered ring; wherein ring A or ring B is attached to group W of any position through group V; wherein V is a chemical bond, or V is a group (CH2) n- (where n "= 1-2), or V is a group C = 0, and wherein Z is selected from the group consisting of: a hydrogen atom, an optionally substituted C 1 -6 alkyl group, an optionally substituted aromatic or heterocyclic group, an optionally substituted amino group, an optionally substituted C 3 -6 alkylamino group or C 3 cycloalkylamino group, and a group substituted carbonyl, or pharmaceutically acceptable acid addition salts thereof, wherein said compound may be in any stereoisomeric form or may be present as a mixture of stereoisomeric forms thereof Compounds having the formula (2A) and (2B), and methods for synthesizing said compounds, are set forth in WO 00/56729, incorporated herein by reference, Other CXCR4 antagonists are compounds of formula (3): or the salts, prodrugs and stereochemical forms thereof, wherein: ring A optionally comprises a heteroatom selected from N, O and S; the dotted lines represent optional unsaturation; R1 is halo, nitro, cyano, optionally substituted hydroxy, optionally substituted thiol, optionally substituted amino, carboxylate, carboxamide, sulfonate, sulfonamide, C2- alkanoyl, alkylsulfonyl or aroyl; R2 and R3 are independently H, an optionally halogenated C? -4 alkyl, an optionally substituted aryl or heterocyclic group, or R2 and R3 together with the E ring can form a substituted or unsubstituted 5 to 7 membered ring; k is 0-4; m is 0-2; L1 is a covalent bond or C? -6 alkyl optionally containing N or O; X is C substituted or unsubstituted, N; u O or S; Ar is phenylene; each n is independently 0-2; each R is independently H or alkyl (1-6 carbons); and Y is a fused or non-fused aromatic or heteroaromatic ring, or a 5-6 membered heterocyclic group. The CXCR4 antagonists can also have the formula (3A): or the salts, prodrugs and stereochemical forms thereof, wherein: R, m, n, Ar and each Y are defined as in formula (3); L2 is a covalent bond or C1-6 alkyl optionally containing N or O; and each Z is independently CR2, NR, O or S, with the proviso that only two Z can be less CR2.

In formula (3A) above, L2 may be methylene or ethylene. In one example, m is 1, and all modes of Z are CR2, in particular CH2. In formula (3A) above, each Y can be pyrimidyl, pyridyl, phenyl, benzimidazole or benzoxazole. Other CXCR4 antagonists have the formula (3B): or the salts, prodrugs and stereochemical forms thereof, wherein: W1 is a monocyclic (5 to 6 membered) or bicyclic (8 to 12 membered) ring system not substituted or substituted containing at least one heteroatom selected from N, O and S; W2 is H, or is selected from the group consisting of: an optionally substituted C? -6 alkyl group; an alkyl group of Co-6 substituted with an optionally substituted aromatic or heterocyclic group; an optionally substituted C3-7 alkylamino group of Co-6 or cycloalkylamine; and an optionally substituted carbonyl or sulfonyl group; Ar, R and n are defined as in formula (3), and is a saturated or unsaturated 5-membered ring containing 1-2 heteroatoms selected from N, O and S. Other CXCR4 antagonists have the formula (3C): or the salts, prodrugs or stereochemical forms thereof, wherein: W1 is phenyl, pyridyl, pyridimyl, imidazolyl, thiophenylyl, and a fused ring system optionally having a heteroatom selected from N, O and S; W2 is H; Ar, R and n are defined as in formula (3); Y represents a 10-membered fused ring system, optionally containing 1 or 2 heteroatoms selected from N, O and S.

Compounds having the formula (3) and (3A) - (3C), and methods for synthesizing said compounds, are set forth in WO 02/22600, which is incorporated herein by reference. Other CXCR4 antagonists have the formula (4): or the salts, prodrugs and stereochemical forms thereof, wherein: X is a monocyclic (5 to 6 membered) or bicyclic (9 to 12 membered) ring system not substituted or substituted containing at least one heteroatom selected from N, O and S; Z is H, or is a 5-6 membered monocyclic ring or fused bicyclic system of 9 to 12 members, optionally substituted, containing N, O or S; Ar is an optionally substituted aromatic or heteroaromatic ring; each of L1, L2 and L3 is independently a link, CO, SO2 or CH2, wherein at least one of L2 and L3 must comprise CO or SO2; and wherein L1 may also be alkylene (2-5 carbons), wherein one or two carbons may be optionally replaced by N, and which alkylene may be optionally substituted by an alkylene bridged (3-4 carbons); L2 and L3 can also be, independently, SO2NH, CONH, S02NHCH2 or CONHCH2; n is 0, 1 or 2; each R1 and R2 is independently H or straight or branched chain or cyclic alkyl (1-6 carbons), which may be optionally substituted, and wherein R2 may be alkylene coupled to Y; and Y comprises at least one aromatic or heteroaromatic ring or other substituted or unsubstituted heterocyclic ring directly coupled to L3. In formula (4) above, X may be dihydroquinoline, tetrahydroquinoline, pyranopyridine, dihydropyranopyridine, thiapranopyridine., dihydrothiapiranopyridine, dihydronaphthyridine, tetrahydronaphthyridine, imidazolyl, oxazolyl, thiazolyl, benzimidazolyl, benzothiazole or benzoxazolyl. In formula (4) above, L1 can be alkylene (2-5 carbons), wherein a carbon can optionally be replaced by N, and which can be optionally substituted by an alkylene bridged (3-4 carbons). For example, L1 may be alkylene, CO or S02, and X is a optionally substituted imidazole, oxazole, thiazole, benzimidazole, benzothiazole or benzoxazole. Alternatively, L1 may be a bond, and X is dihydroquinoline, tetrahydroquinoline, pyranopyridine, dihydropyranopyridine, thiapyranopyridine, dihydrotiapyranopyridine, dihydronaphthyridine or substituted or unsubstituted tetrahydronaphthridine. In formula (4) above, Z can be hydrogen.

In formula (4) above, Y may be an imidazole, benzimidazole, pyridine, pyrimidine or optionally substituted phenyl, wherein the ring nitrogen may optionally be oxidized. For example, Y can be substituted with halogen, nitrile, alkyl, -OR, -SR, -NR2, -NRCOR, -OOCR, -COR, -CONR2, -COOR, -N02, -NOH or -CF3, wherein R is H or alkyl (1-6 carbons). In the above formula (4), each X or Z may be optionally substituted by halo, nitro, cyano, carboxy, C-MO alkyl, C2-0 alkenyl, C3.10 cycloalkyl, hydroxy, thiol, amino, acyl , carboxylate, carbamate, carboxamide, sulfonamide, a carbonyl or sulfonyl which is bonded to a hydrogen, or substituted with a C - ?. alquilo alkyl, C al-? alkenyl or C3-7 cycloalkyl or a monocyclic aromatic group of 5 to 6 members; or X or Z may be optionally substituted by a 5- to 6-membered monocyclic aromatic group, naphthyl or a 5- to 6-membered heterocyclic ring. Other CXCR4 antagonists have the formula (4A): (4A) or the formula (4B): where 1 is 0-3, and R 'is OH, MeO, SH SMe, CN, C02Me, F, Cl, Br, N02, CH3CO, NH2, NHCH3, N (CH3), CH3CONH, CH3S02NH, CONH2, S02NH2, CF3 or Me; each of Z1, Z2 and Z3 is independently CH, CR 'or N, wherein only two of said Z1, Z2 and Z3 can be N; and L2 and L3 are as defined in formula (4). In formula (4A) or (4B) above, Z1, Z2 and Z3 can be CH or CR '. In one example, Z3 is N and L3 is CO. In addition, one of L2 and L3 can be S02, and the other is a link or CH2. Alternatively, one of L2 and L3 is CO, and the other is a bond or CH2. In another embodiment, the compound for use in the methods of the present invention has the formula (4C): where 1 is 0-3, and R 'is OH, MeO, SH SMe, CN, C02Me, F, Cl, Br, N02, CH3CO, NH2, NHCH3, N (CH3) 2, CH3CONH, CH3S02NH, CONH2, S02NH2 , CF3 or Me; k is 0-2; each of Z1, Z2 and Z3 is independently CH, CR 'or N, wherein only two of said Z1, Z2 and Z3 can be N; and X, L2 and L3 are as defined in formula (4). In formula (4C) above, Z1, Z2 and Z3 can be CH or CR '. In one example, Z3 is N and L3 is CO. In addition, one of L2 and L3 can be S02, and the other is a link or CH2. Alternatively, one of L2 and L3 may be CO, and the other is a bond or CH2. Compounds having the formula (4) and (4A) - (4C), and methods for synthesizing said compounds, are set forth in WO 02/22599, which is incorporated herein by reference. Other CXCR4 antagonists have the formula (5): the salts, prodrugs and stereoisomeric forms thereof; Ring A optionally comprises a heteroatom selected from N, O and S; dotted lines represent optional unsaturation; R1, R2 and R3 are independently H, halo, alkyl, hydroxyl, amine, thiol or substituted or unsubstituted acyl; or R2 and R3 can together form a benzo ring; k is 0-4; 1 is 0, 1 or 2; X is C or N substituted or unsubstituted; or is O u S; Ar is the residue of an aromatic or heteroaromatic portion; each n is independently 0-2; each R is independently H or alkyl (1-6 carbons); j is 0-3; and each Y is independently selected from the group consisting of halo, OR; SH; SW; S02; optionally substituted phenyl; - (CR2) mOR; - (CR2) mCOR; - (CR2) mCOOR; - (CR2) mN = CH- NR2; - (CR2) mCONHNHR; - (CR2) mCN; - (CR2) mNR52; - (CR2) mNR (CR2) mNRR4; - (CR2) mNR (CR2) mNR (CR2) mNR52; - (CR2) mCO (CR2) mNR52; - (CR2) mCO (CR2) mNR (CR2) mNRR4; - (CR2) mCO (CR2) mNR (CR2) mNR (CR2) mNR52; - (CR2) mNRCO (CR2) mNRR4; - (CR2) mNRCO (CR2) mNR (CR2) mNR52; - (CR2) mNRCO (CR2) mNR (CR2) mNR (CR2) mNR (CR2) mNR52; - (CR2) mNROH; - (CR2) mCONROH; - (CR2) mCR = NOH; - NHNHR; - CH = N-Z; and - guanidino or amidino, each of which can be linked to Y through a portion (CR2) m; wherein R is H or alkyl (1-6 carbons), each m is independently 0-4, and each R4 and each R5 is independently H, alkyl (1-6 carbons), alkenyl (2-6 carbons), alkynyl ( 2-6 carbons) or acyl (1-6 carbons), each optionally substituted by one or more non-aromatic or non-heterocyclic substituents, wherein two R5 may be attached to form a cyclic amine optionally containing one or more additional heteroatoms selected of N, O and S; a indicates the linker between ring A and N; b indicates the linker between ring E and N; and wherein Z is an aromatic or heteroaromatic portion containing 5-12 ring members. In the above formula (5), Ar can be a 5 to 6 membered monocyclic ring or a 9 to 12 membered fused ring system. For example, Ar can be benzene, naphthalene, dihydronaphthalene, tetrahydronaphthalene, pyridine, pyrimidine, quinoline, isoquinoline, midazole, benzimidazole, azabenzimidazole, benzotriazole, furan, benzofuran, thiazole, benzothiazole, oxazole, benzoxazole, pyrrole, indole, midazole, tetrahydroquinoline, tetrahydroisoquinoline, pyrazole, thiophene, isoxazole, isothiazole, triazole, tetrazole, oxadiazole, thiadiazole, imidazoline and benzopyran. In particular examples, Ar is benzene, benzimidazole, benzothiazole, imidazole, oxazole, benztriazole, thiazole, pyridine or pyrimidine. In one embodiment, at least one Y is - (CR2) mNR52. In formula (5) above, R2 and R3 can together form a benzo substituent. In another embodiment, X is N and ring E comprises a pi bond coupled to an N. In one embodiment, ring E is coupled to the remainder of the molecule at position 2. In formula (5) above, ring A it can be saturated and 1 is 1. In an example, k is 0-1. In other examples, the ring system including A is tetrahydroquinoline or a substituted form thereof. In formula (5) above, one of (CR2) an and (CR2) bn can be CH2, and the other is a link. For example, (CR2) an can be a bond, and (CR2) bn is CH2. Compounds having the formula (5), and methods for synthesizing said compounds, are set forth in WO 02/34745, which is incorporated herein by reference. Other CXCR4 antagonists have the formula (6): (6) or the salts, prodrugs and stereoisomeric forms thereof, wherein X and Y are independently N or CR1; Z is S, O, NR1 or CR12; each R1-R6 is independently H, halo, 0 (C = 0) R, NR (C = 0) R, OR, SR, NR2, COOR, CONR2, wherein R is H or optionally substituted alkyl, alkenyl, alkynyl or aryl; or each R1-R6 is alkyl (C? -? o), alkenyl (C2-? o). alkynyl (C2 -? 0), aryl (05-12), arylalkyl, arylalkenyl or arylalkynyl, each being optionally substituted and optionally containing O, S or N; or an optionally substituted acyl, arylacyl, alkyl-alkenyl-, alkynyl- or arylsulfonyl, wherein each alkyl, alkenyl, alkynyl or aryl portion may contain O, O or N; n1 is 0-4; n2 is 0-1, where * means C = C, and can be replaced by CR5 = CR5; n3 is 0-4; where n1 + n2 + n3 is greater than or equal to 2; b is 0-2; wherein the following combinations of R groups can be coupled to generate a ring, whose ring can be saturated or unsaturated: R + R2 an R2 + R3 R3 + an R4, R + R4, an R5 + another R5, an R5 + an R6 , and R6 + R6; wherein the ring may not be aromatic when the participants in the formation of the ring are two R5; and where when n2 is 1, neither n1 nor n3 can be 0. Other CXCR4 antagonists have the formula (6A): or the salts, prodrugs and stereoisomeric forms thereof, wherein R1-R6 and n1-n3 are as defined in formula (6).

Other antagonists have the formula (6B) or the formula (6C): (6B) (6C) or the salts, prodrugs or stereoisomeric forms thereof, wherein n is 0-1; d is 0-3; the dotted line is an optional F link; and R1-R6 are defined as in formula (6). In another embodiment, the compounds for use in the methods of the present invention have the formula (6D): (6D) or the salts, prodrugs and stereoisomeric forms thereof, wherein R -R are defined as in formula (6), and n4 is 2-6.

In the formula (6) or (6A) - (6D) above, each R1 can be H, halo, alkyl, alkoxy or CF3. In one embodiment, each R2 is H or alkyl. In another embodiment, each R3 is H, alkyl, alkenyl, arylalkyl or aryl. In the formula (6) or (6A) - (6D) above, each R 4 can be H, alkyl or aryl. Alternatively, two R4 may form an optionally substituted aromatic or aromatic ring heterocycle. For example, two R 4 can form a phenyl or pyridyl ring, which can be substituted with halo, alkyl, halogenated alkyl, hydroxy or alkoxy. In the formula (6) or (6A) - (6D) above, each R 5 can be H, alkyl or alkenyl, wherein said alkyl or alkenyl can be optionally substituted. In one embodiment, the alkyl or alkenyl substituents on a single carbon, or on adjacent or non-adjacent carbons, form a saturated or unsaturated ring. In one example, the substituents form a non-aromatic ring. In another embodiment, an R5 is an oxime, an alkylated oxime, alkylated hydroxylamine, hydroxylamine or halo. In the formula (6) or (6A) - (6D) above, each R6 may independently be H or an arylalkyl or arylsulfonyl, wherein the aryl portion may comprise a heteroatom; or two R6 may comprise a guanidyl, carbonyl or carbamino group. In one embodiment, two R6 together, or one R5 and one R6 may together form a saturated or unsaturated aromatic ring, wherein each ring may optionally contain N, S or O.

Compounds having the formula (6), and methods for synthesizing said compounds, are set forth in WO 03/055876, which is incorporated herein by reference. The CXCR4 antagonists can have the formula (7): or the salts, prodrugs or stereoisomeric forms thereof, wherein X is (CR32) 0 - (CR3 = CR3) P - (CR32) q - NR52; (CR32), R4; or an optionally substituted benzyl, or a monocyclic or bicyclic ring optionally containing N, O or S; Y is an optionally substituted 5- to 12-membered heterocyclic ring containing a nitrogen atom, said heterocyclic ring may be monocyclic or fused, and is aromatic or partially aromatic; A and R1 are independently halo, CF3, cyano, nitro, OR, SR, NR2, COOR, CONR2, NS02R, OS02R3 or OS02NR, wherein each R is H, alkyl, alkenyl, alkynyl or aryl; or A and R1 are independently an alkoxy (C-MO), alkyl (C-MO), alkenyl (C2-? o), alkynyl (C2-? 0), aryl (5-12 members), arylalkyl, arylalkenyl or arylalkynyl optionally substituted, each of which may optionally contain O, S or N; R2 and R3 are independently H or an optionally substituted alkyl; R4 is an optionally substituted heterocyclic ring or heteroaryl; or R4 comprises a portion urea, hydroxyurea, sulfamide, acetamide, guanidine, cyanamide, hydroxylamine, cyanamide, imidazolidin-2-one or nicotinamide, each of which can be substituted with a heterocyclic ring; R5 is H or alkyl; 1 and n are independently 0-4; p is 0-1; or and q are independently 1-4; and r is 1-6. In formula (7) above, at least one of R and R2 may not be H, and may be linked to form an additional ring such as an aryl or heteroaryl. In one example, two A's may not form an additional ring. In another example, X is (CR32) r -R4, r is at least two, and R4 is 2-pyridinyl, quinolinyl, imidazolyl or furan. In formula (7) above, X can be (CR32) 0 - (CR3 = CR3) P - (CR32) q - NR52, where each R3 and R5 are independently H, and p can be zero. In particular modalities, o and q are together 2-6. Alternatively, X may be (CR32) r -R4, where R4 is a heterocyclic or heteroaryl ring, each of which contains a nitrogen atom. For example, R 4 can be azetidine, pyrrolidinyl, pyridinyl, thiophenyl, imidazolyl or benzimidazolyl. Alternatively, X may be a monocyclic or bicyclic ring optionally containing N, O or S, such as cyclohexyl, piperidine, 8-aza-bicyclo [3.2.1] octane or 3-aza-bicyclo [3.2.1] octane . In another embodiment, X is an optionally substituted benzyl, in particular a disubstituted benzyl. In the above formula (7), Y may be a 5- to 6-membered heterocyclic ring containing a nitrogen atom adjacent to the atom that is attached to the rest of the molecule. The 5-6 membered heterocyclic ring may be fused to another ring. For example, Y can be pyridine, pyrimidine, pyrazine, indole, benzimidazole, benzothiazole, imidazole, isoquinoline, tetrahydroquinoline, pyridazine, thiazole or benzoimidazole. In particular examples, Y is tetrahydroquinoline, in particular a 5,6,7,8-tetrahydroquinoline moiety, linked at position 8 to the rest of the molecule. In the above formula (7), each optionally substituted portion can be substituted with a heteroatom, halo, CF3, cyano, nitro, hydroxy, alkoxy, carbonyl, carboxy, amino, amido, imino, cyano, sulfonyl; C 2-6 alkyl or C 2-6 alkenyl, each of which may contain N, O or S; or it can be substituted with an aryl, hateroaryl, carbocyclic or heterocyclic ring, each of which can be further substituted with the same substituents. Compounds having the formula (7), and methods for synthesizing said compounds, are set forth in WO 04/091518, which is incorporated herein by reference.

The CXCR4 antagonists can have the formula (8): Or the salts, prodrugs and stereoisomeric forms thereof, wherein each of the rings A and B is independently an optionally substituted 5 to 6 membered monocyclic heteroaryl; Ring C is an optionally substituted, saturated or partially saturated 5 to 7-membered ring, and may contain a heteroatom in addition to nitrogen, wherein said heteroatom is N, O or S; Y is H, a C? -6 alkyl containing one or more heteroatoms, or a cyclic portion, each of which is optionally substituted; R1 and R2 are independently H, halo or an optionally substituted alkyl; L is (CR32)? or NR (CR32) ?, wherein an alkyl bond can be replaced with an alkenyl or alkynyl bond; 1 is 1-6; and each R3 is H or alkyl.

In formula (8) above, at least one of R1 and R2 may not be H when C is piperidinyl or 1, 2,3,6-tetrahydropyridinyl, and rings A and B are pyridinyl. In other embodiments, R1 and R2 are not both naphthalenyl when ring C is piperidinyl and rings A and B are pyridinyl. In other embodiments, ring C is not 4-oxo-piperidine-3,5-dicarboxylic acid if L-Y is CH 3; and ring C is not 4-hydroxy-1, 2,5,6-tetrahydro-pyridine-3-carboxylic acid ester if LY is benzyl. In formula (8) above, R1 and R2 may be at positions adjacent to the bonds with ring C. In one example, R1 and R2 are independently unsubstituted alkyl, such as methyl. In the above formula (8), each of the rings A and B can be pyridine, pyrimidine, pyrazine, pyridazine, 1,2,3-triazine, 1,4-triazine, 1, 3,5-triazine, 1, 2,4,5-tetrazine, pyrrole, imidazole, pyrazole, 1,2-triazole, 1,4-triazole, tetrazole, thiazole, oxazole, isothiazole, isoxazole, 1,2-thiadiazole, 1, 3,4-thiazole, 1, 2,3-oxadiazole, 1,4-oxadiazole, quinoline, isoquinoline, quinoxaline, quinazoline, phthalazine, cinnoline, 1,3-benzotriazine, 1,4- benzotriazine, indole, benzimidazole, 1 H-indazole, benzoxazole, benzthiazole, benz [d] isoxazole, benz [c sotiazole or purine. In particular examples, each of the rings A and B is pyridine, pyrimidine, midazole or benzimidazole, and each of the rings A and B may be identical. Each of the rings A and B may also contain a single substituent, which may be identical, in the position adjacent to the bond joining the rings to the ring C.

In formula (8) above, ring C can be a saturated ring, or it can contain a double bond. For example, ring C can be pyrrolidine, piperidine, hexahydro-1 / - / - azepine, piperazine, morpholine, thiomorpholine, azepam, azocan, 2,3,4,7-tetrahydro-1 / - / - azepine, 2, 3,6,7-tetrahydro-1H-azepine, 3-pyrroline, 1, 2,3,6-tetrahydropyridine, isoindoline, 1, 2,3,4-tetrahydroisoquinoline, 2,3,4,5-tetrahydro-1 H -benzo [a (] azepine, 2,3,4,5-tetrahydro-1 H -benzo [c] azepine, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclopentene, cyclohexene, cycloheptene, cyclooctene, tetrahydropyran, tetrahydrotiopivane, oxepane , tiepano, oxocano or thiocano In particular examples, the C ring is pyrroline, piperidine, piperazine or hexahydro-1H-azapine The C ring can be substituted with an alkyl, halo, cyano, oxime, OR or C = N-OR optionally substituted, wherein R is an optionally substituted alkyl In formula (8) above, Y may be selected from the group consisting of: - (CR2) mNR2, - (CR2) mNR2 (CR3), - (CR2) mNR ( CR2) mNR2, - (CR2) mNR (CR2) mNR (CR2) mNR2, - (CR2) m OR, - (CR2) mCO (CR2) mOR, - (CR2) mCO (CR2) mNR2, - (CR2) mCO (CR2) mNR (CR2) mNR2, - (CR2) mNRCO (CR2) mNR2, - (CR2) mNR (CR2) mC02R, - (CR2) mNR (CR2) mCOR, - (CR2) mNR (CR2) mS02R, - (CR2) mNRCO (CR2) mNR (CR2) mNR2, - (CR2) mNRCO (CR2) mNR ( CR2) mNR (CR2) mNR (CR2) mNR2, - (CR2) mNR (CR2) mOR, - (CR2) mCR = NOH, - (CR2) mCONR (CR2) mOR, - (CR2) mN [(CR2) mC02R ] 2, - (CR2) mONRCONR2, - (CR2) mZ, - (CR2) mNR- (CO) mZ, - (CR2) mNR- (CR2) mZ and - (CR2) m-CR = N = Z; wherein each R is H or an optionally substituted alkyl, each m is independently 0-4; and Z is an optionally substituted aromatic or heteroaromatic portion containing from 5 to 12 ring members. In particular modalities, Y is (CH2)! NR2 and 1 is 1-10. Alternatively, Y may be an aromatic, heteroaromatic or heterocyclic portion of 5 to 12 members, each of which may be a monocyclic or fused ring. For example, Y can be phenyl, imidazole, pyridine, thiophene, pyrrolidine, pyrazole, piperidine, azetidine, benzimidazole, benzota soxazole or thiazole. In addition, Y can be optionally substituted with halo; cyano; nitro; alkoxy; halogenated alkyl; substituted carbonyl; a cyclic portion such as an aryl or heteroaryl of 5 to 12 members containing N, O or S; or an alkyl, alkenyl, or a heteroalkyl portion optionally containing one or more N, O, S, each of which is optionally substituted and is optionally in the form of oxides. In particular examples, Y is substituted with pyridine, phenyl, piperidine or 2H-tetrazole. In the above formula (8), each optionally substituted group can be substituted with inorganic portions such as a heteroatom, halo, nitro, hydroxy, carboxy, amino, amido, cyano or sulfonyl; or it can be substituted with alkyl (C-MO), alkenyl (C2-? o), alkynyl (C2-? 0), aryl (5-12 members), arylalkyl, arylalkenyl and arylalkynyl, each of which can contain optionally a heteroatom such as O, S or N, and each of which can be further substituted with the same substituents. For example, each optionally substituted alkyl may be substituted with a heteroatom such as N, O or S, or with a carbocyclic, heterocyclic, aryl or heteroaryl substituent. Compounds having the formula (8), and methods for synthesizing said compounds, are set forth in WO 04/093817, and in the patent application of E.U.A. published as US 2005/0154201, each of which is incorporated herein by reference.

It is understood that the above detailed description and the accompanying examples are illustrative only, and will not be considered as limitations on the scope of the invention. The publications and patents of E.U.A. referred to herein are incorporated herein by reference.

Claims (13)

NOVELTY OF THE INVENTION CLAIMS

1. The use of at least one CXCR4 antagonist in the preparation of a pharmaceutical composition useful for improving the efficacy of a chemotherapeutic method in an affected subject with a hematopoietic or myeloid malignancy.

2. The use as claimed in claim 1, wherein the CXCR4 antagonist is of the formula: Z-linker-Z '(1), or pharmaceutically acceptable salt or prodrug forms thereof, wherein Z is a cyclic polyamine containing 9-32 ring members of which 2-8 are nitrogen atoms, said nitrogen atoms being separated from each other by at least 2 carbon atoms, and wherein said heterocycle optionally may contain additional heteroatoms in addition of nitrogen, and / or may be fused to an additional ring system; or Z is of the formula: A \ N / B wherein A comprises a monocyclic or bicyclic fused ring system containing at least one N, and B is H or an organic portion of 1-20 atoms; Z 'may be described in a form as defined by the above Z, or may alternatively be of the formula: -N (R) - (CR2) n-X, wherein each R is independently H or straight, branched or cyclic alkyl (1-6 carbons), n is 1 or 2, and X is an aromatic ring, including heteroaromatic rings, or is a mercaptan, or Z 'is absent; "linker" represents a bond, alkylene (1-6 carbons), or may comprise aryl, fused aryl, oxygen atoms contained in an alkylene chain, or may contain keto groups or nitrogen or sulfur atoms.

3. The use as claimed in claim 2, wherein Z and Z 'are both cyclic polyamines.

4. The use as claimed in claim 3, wherein the compound of formula (1) is 1,1 '- [1,4-phenylene-bís- (methylene) -bis- 1, 4,8 , 11-tetraazacyclotetradecane (AMD3100).

5. The use as claimed in claim 2, wherein Z is a cyclic polyamine, and Z 'is N (R) - (CR2) n-X.

6. The use as claimed in claim 5, wherein each R is H, n is 2, and X is substituted or unsubstituted pyridyl.

7. The use as claimed in claim 6, wherein the compound of formula 1 is N- [1,4,8,11-tetraazacyclotetradecanyl- (1,4-phenylene-bis- (methylene)] - 2-aminoethyl-2-pyridine (AMD3465)

8. The use as claimed in claim 2, wherein Z is of the formula: A \ N / B wherein A comprises a monocyclic or bicyclic fused ring system containing at least one N, and B is H or an organic portion of 1 to 20 atoms.

9. The use as claimed in claim 8, wherein A is 5,6,7,8-tetrahydroquinolin-8-yl, and B is 1 H-benzimidazol-2-ylmethyl.

10. The use as claimed in claim 9, wherein Z 'is absent, and the linker is an aminoalkyl omega substituent.

11. The use as claimed in claim 10, wherein the compound of formula 1 is N1- (1 H-benzimidazol-2-ylmethyl) -N1- (5,6,7,8-tetrahydroquinolin) -8-yl) butane-1,4-diamine (AMD11070).

12. The use as claimed in any of claims 2 to 11, wherein the pharmaceutical composition is adapted to be administrable in a dosing scale of approximately 0.1 μg / kg-5 mg / kg of body weight.

13. A pharmaceutical composition comprising an effective amount of a CXCR4 antagonist in a unit dosage form for improving the efficacy of a chemotherapeutic method in a subject.