HK1158549B - Treatment of cancers of the blood using selected glycomimetic compounds - Google Patents

Description

Treatment of hematologic cancers using selected glycomimetic compounds

Background

Technical Field

The present invention relates generally to methods for treating hematological cancers or complications associated therewith, and for reducing myeloablative (myeloablative) bone marrow toxicity of chemotherapy regardless of cancer type, and more particularly to the use of specific glycomimetics for treatment.

Description of the Related Art

One of the cancer groups is hematological cancer. Such cancer groups include hematologic malignancies. Acute myelogenous leukemia is an example of such a hematological cancer.

Acute myeloid leukemia (also known as acute myeloid leukemia or AML) is a cancer of the white blood cells, especially of the myeloid lineage. It appears that AML is caused by a single progenitor cell that has undergone genetic transformation into an abnormal cell with rapid proliferative capacity. These abnormal immature bone marrow cells accumulate in the bone marrow. This accumulation in the bone marrow interferes with the production of normal blood cells, including a decrease in red blood cells, platelets, and neutrophils. Eventually the bone marrow ceases to function properly.

AML is one of the most common types of leukemia in adults, and is the most common acute leukemia affecting adults. In the united states alone, there are about 12,000 new cases per year. The incidence of AML is expected to increase with the age of the population. In addition, in the united states, about 11% of childhood leukemia cases are AML. Chemotherapy is commonly used to treat AML. Current treatments cure only a few patients.

Chemotherapy has a number of deleterious side effects. One such side effect is myeloablative (myeloablative) bone marrow toxicity. Bone marrow is a tissue that fills the interior of some bones. Examples of such bones are the sternum, hip, femur and humerus. Bone marrow contains stem cells that develop into several cell types: erythrocytes (erythrocytes), leukocytes (leukocytes), and thrombocytes (platelets). Due to their rapid division rate, cells in the bone marrow are susceptible to chemotherapy. The bone marrow is prevented from forming new blood cells by the chemotherapeutic agent. Over time following exposure to the chemotherapeutic agent, the count of blood cells will decline at different rates depending on the particular type of cell as their average life span varies. For example, a low white blood cell count makes an individual more susceptible to infection. For example, a low red blood cell count causes the individual to experience fatigue. For example, a low platelet count impairs the ability of an individual to produce blood clots.

Accordingly, there is a need in the art to treat hematological cancers, including acute myeloid leukemia, or complications associated therewith, as well as to reduce myeloablative myelotoxicity of chemotherapy. The present invention fulfills these needs and provides other related advantages.

Summary of The Invention

Briefly, the present invention provides methods for treating hematologic cancers or complications associated therewith, as well as cancer-type independent methods for reducing myeloablative bone marrow toxicity of chemotherapy. In the present invention, the compounds for treatment or reduction comprise or consist of specific glycomimetics. Such compounds may be combined with a pharmaceutically acceptable carrier or diluent to form a pharmaceutical composition.

In one embodiment, the present invention provides a method for treating a hematologic cancer or a complication associated therewith in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound having the formula:

wherein L ═ linker group; and n is 0-1

In one embodiment, the present invention provides a method for reducing myeloablative bone marrow toxicity of chemotherapy in a subject in need of such reduction, said method comprising administering to said subject an amount effective for said reduction of a compound having the formula:

wherein L ═ linker group; and n is 0-1

In one embodiment, the above compound is combined with a pharmaceutically acceptable carrier or diluent.

In one embodiment, the hematologic cancer is Acute Myeloid Leukemia (AML).

In other embodiments, the above-described compounds or compositions thereof may be used for the manufacture of a medicament for any of the uses described herein.

These and other aspects of the invention will become apparent by reference to the following detailed description and attached drawings. All references disclosed herein are incorporated by reference in their entirety as if each were individually incorporated.

Drawings

FIG. 1 is a diagram illustrating the compositional synthesis of Compound #1(Compound # 1).

FIGS. 2A-2C are diagrams illustrating the compositional synthesis of Compound # 1.

FIG. 3 is a diagram illustrating modification of the components of FIG. 1.

Figure 4 is a diagram illustrating the reaction of the components of figures 2 and 3 to form compound # 1. Compound XIX of fig. 2 is reacted with Ethylenediamine (EDA) to form EDA-XIX.

FIG. 5 is a diagram illustrating the synthesis of Compound #2(Compound # 2). Compound XIX of figure 2 is reacted with ethylenediamine to form EDA-XIX.

FIG. 6 shows that Compound #2 inhibits E-selectin and P-selectin AML rolling on human endothelial cells.

FIG. 7 shows the effect of Compound #2(1.5mM) on the adhesion of U266 myeloma cells to P-selectin expressing endothelial cells in the flow state.

Figure 8 shows the effect of compound #2 (at different concentrations) on SDF-1-induced transendothelial migration of Multiple Myeloma (MM) cells.

FIG. 9 shows the effect of Compound #2(25mg/kg or 100mg/kg) on extravasation of blood stream from multiple myeloma cells.

Fig. 10 is a schematic of an experimental design investigating the effect of compound #2 on chemotherapy-induced neutropenia.

Figure 11 shows the effect of compound #2 on neutrophil recovery in mice treated with cyclophosphamide.

FIG. 12 shows the effect of Compound #2 on neutrophil recovery in mice treated with 5-fluorouracil (5-FU).

Detailed Description

As described above, the present invention provides methods for treating hematological cancers or complications associated therewith, as well as methods for reducing myeloablative bone marrow toxicity of chemotherapy regardless of cancer type.

Compounds useful in compositions (including pharmaceuticals) and methods of the invention include embodiments having the formula:

in the above formula, "L" represents a linker. There may be no linker present (i.e., "n" is 0) or there may be a linker present (i.e., "n" is 1). If no linker is present, the compound has the formula:

if n is 1, a linker is present. The linker may be (or may include) a spacer group, such as- (CH)₂)_P-or-O (CH)₂)_PWherein p is generally from about 1 to about 20 (including any whole integer range therein). Examples of other spacer groups include carbonyl groups or carbonyl groups containing groups such as amides. One embodiment of such a spacer group is

It produces:

embodiments of linkers include the following:

other linkers, e.g. polyethylene glycol (PEG) or-C (═ O) -NH- (CH)₂)_P-C(＝O)-NH₂Wherein p is as defined above, are familiar to those skilled in the art or are within the present disclosure.

In other embodiments, the linker is

It produces:

in other embodiments, the linker is

It produces:

all compounds of the invention or useful therefor (e.g. pharmaceutical compositions or methods of treatment) include physiologically acceptable salts thereof. Examples of such salts are sodium, potassium, lithium, magnesium, calcium and chlorine.

The compounds described herein may be present within a pharmaceutical composition. Pharmaceutical compositions comprise one or more compounds in combination (i.e., not covalently bound) with one or more pharmaceutically or physiologically acceptable carriers, diluents, or excipients. Such compositions may comprise a buffer (e.g., a neutral buffer salt or phosphate buffer salt), a carbohydrate (e.g., glucose, mannose, sucrose, or dextran), mannitol, a protein, a polypeptide, or an amino acid such as glycine, an antioxidant, a chelator such as ethylenediaminetetraacetic acid (EDTA) or glutathione, an adjuvant (e.g., aluminum hydroxide), and/or a preservative. In still other embodiments, the compositions of the present invention may be formulated as a freeze-dried product. The compositions of the invention may be formulated for appropriate modes of administration including, for example, topical, oral, nasal, intravenous, intracranial, intraperitoneal, subcutaneous, or intramuscular administration.

The compositions described herein may be administered as part of a sustained release formulation (i.e., a formulation such as a capsule or sponge that causes the compound to be slowly released upon administration). Such formulations are generally prepared using well known techniques and are administered, for example, by oral, rectal or subcutaneous implantation, or by implantation at the desired target site. The carriers used in such formulations are biocompatible and may also be biodegradable; preferably the formulation provides a relatively constant level of release of the compound. The amount of compound included in the sustained release formulation depends on the site of implantation, the rate and expected duration of release, and the nature of the disease state to be treated or prevented.

The compounds described above, including equivalents thereof, are useful in the methods of the invention because they are associated with hematological cancers. Hematologic cancers include hematologic malignancies. Examples of hematological cancers include Acute Myelogenous Leukemia (AML), Acute Lymphoblastic Leukemia (ALL), Chronic Myelogenous Leukemia (CML), and Multiple Myeloma (MM). In one embodiment, a therapeutically effective amount of at least one (i.e., one or more) of the above-described compounds is administered to an individual in need of treatment of a hematologic cancer or a complication associated therewith. As used herein, the term "treatment" (including variants such as "treating") includes prophylaxis. For example, complications associated with hematologic cancer may not manifest themselves in an individual suffering from the disease, and a compound may be administered to prevent the occurrence of the complications in the individual. Complications associated with hematologic cancers include, for example, shortened life expectancy, organ damage, periodic or chronic pain, migration of cancer cells out of the blood circulation, and reduction of red blood cells, white blood cells, or platelets. It is desirable to prevent cancer cells from leaving the primary site or from escaping the blood stream and penetrating into other tissues. Cancer cells are usually sensitive to chemotherapy while in the bloodstream, but once they leave the bloodstream, treatment is much more difficult. For example, cancer cells (such as MM cells) can spill from the blood and infiltrate the bone marrow stroma where they do not access the circulating chemotherapeutic agent in the blood stream. Complications of the migration of cancer cells out of the blood circulation include relapse (treatment failure) and disseminated disease (metastasis) leading to organ damage or failure, for example. AML is an example of a hematologic cancer with complications of the migration of cancer cells out of the blood circulation, which lead to disseminated disease.

As noted above, the term "treatment" refers to any of a number of positive effects brought about by treatment, including, for example, elimination of complications associated with the disease, alleviation of complications to some extent, slowing or stopping disease progression, and prolonging the survival time of the patient. Such treatment may be used in conjunction with one or more other treatments for hematological cancers or complications associated therewith. The use of such treatment in combination with another treatment may provide two treatments, each of which exert their own effect to treat the cancer or complications associated therewith; or two treatments may be provided, one of which enhances the efficacy of the other (e.g., increases the efficacy of the other or improves the outcome of the other) to treat the cancer or complications associated therewith. Such treatment may be useful, for example, to prevent or reduce the migration of cancer cells out of the blood circulation. By acting to retain cancer cells in the bloodstream, this treatment enhances the efficacy of another co-used therapeutic approach and works by providing chemotherapeutic agents to the bloodstream.

The compounds described above, including equivalents thereof, are useful in the methods of the invention as it relates to reducing myeloablative myelotoxicity of chemotherapy independent of cancer type. This applies to, but is not limited to, hematological cancers. Examples of toxicity include low white blood cell count (e.g., low neutrophil count), low red blood cell count, and low platelet count. In one embodiment, at least one compound described above in an amount effective for said reducing is administered to an individual in need of reducing myeloablative bone marrow toxicity of chemotherapy. As used herein, the term "reducing" (including variants such as "reduction") includes partial and total reduction of myeloablative bone marrow toxicity of at least one (i.e., one or more) chemotherapy; but also partial and total prevention of at least one such toxicity (e.g., by administration of at least one compound described above prior to, concurrently with, or subsequent to initiation of chemotherapy). For example, such compounds may not prevent neutropenia, but may promote faster and sustained recovery of neutrophils following chemotherapy.

The compounds described above may be administered in a manner appropriate to the disease to be treated. Appropriate dosages and appropriate durations and frequencies of administration may be determined by factors such as the state of the patient, the type and severity of the patient's disease, and the method of administration. In general, appropriate dosages and treatment regimens provide the compound in an amount sufficient to provide a therapeutic and/or prophylactic benefit. In a particularly preferred embodiment of the invention, the compounds may be administered in a single or multiple daily dosage regimen in a dosage range of from 0.001 to 1000mg/kg body weight (more usually 0.01 to 1000 mg/kg). Appropriate dosages can generally be determined using experimental models and/or clinical trials. In general, it is preferred to use the minimum dose sufficient to provide effective treatment. The efficacy of a patient's treatment is typically monitored using assays appropriate for the condition being treated, which assays are familiar to those skilled in the art.

At least one (i.e., one or more) of the compounds described above can be used in combination with at least one (i.e., one or more) chemotherapeutic agent. The compound may act independently of the chemotherapeutic agent or may act synergistically with the chemotherapeutic agent, for example by enhancing the efficacy of the chemotherapeutic agent or vice versa. In addition, administration can be performed with one or more treatments used to reduce the toxicity of chemotherapy. For example, at least one (i.e., one or more) agent may be administered to counteract (at least in part) chemotherapy side effects. Drugs (chemical or biological) that promote blood cell recovery or enhancement are examples of such agents. The at least one compound described herein may be administered before, after, or simultaneously with the administration of the at least one chemotherapeutic agent or the at least one agent that reduces the side effects of chemotherapy. If administered simultaneously, this combination may be administered from a single container or from two (or more) separate containers.

The following examples are offered by way of illustration and not by way of limitation.

Examples

Example 1: synthesis of BASA (FIG. 1)

Synthesis of Compound 4:3-Nitro-benzyl iodide (1) (48.3g) was added to a commercial aqueous solution (pH 11), 8-aminonaphthalene-1, 3, 5-trisulfonic acid (2) (29.5g), stirring at Room Temperature (RT). The pH of the solution was adjusted to 1 and after evaporation of the solvent, product 3(6.4g) was precipitated from ethanol.

Platinum-catalyzed hydrogenation of compound 3 gave compound 4 (benzylaminosulfonic acid or "BASA" in fig. 1) in 96% yield.

Example 2: synthesis of Glycomimetics (FIG. 2)

Intermediate (II)Synthesis of form II:methanol (MeOH) (200ml) and (-) -shikimic acid (20g) in sulfuric acid (2ml, 98%) were stirred at room temperature for 50 h. The reaction mixture was neutralized with 2N aqueous sodium hydroxide solution in a cooled state. After evaporation to dryness, the residue was purified by silica gel chromatography to afford II (19.2 g).

Synthesis of intermediate III: methylshikimic acid (II, 10g), 2-dimethoxypropane (10ml) and p-toluenesulfonic acid (p-TsOH) (0.8g) were dissolved in acetonitrile (125ml) and stirred at RT for 1 h. The reaction mixture was then neutralized with triethylamine (2ml) and evaporated to dryness. The residue was chromatographed on silica gel to give III (11 g).

Synthesis of intermediate IV: shikimic acid derivative III (10g) and PtO in methanol (40ml) were allowed to stand vigorously with stirring at room temperature₂Hydrogenation of C (10%, 250 mg). After 16 h, the reaction mixture was filtered through celite (celite) and evaporated to dryness. The residue was chromatographed on silica gel to give IV.

Synthesis of intermediate V: pyridine (12ml), acetic anhydride (7ml) and DMAP (25mg) were added to a solution of IV (8g) in DCM (100ml) at 0 ℃. The reaction mixture was stirred at room temperature for 1 h, then diluted with ethyl acetate (EtOAc) (250 mL). After washing with 0.5M aqueous hydrochloric acid (3X 50ml), saturated potassium bicarbonate solution (3X 50ml) and brine (3X 50ml), the combined organic layers were dried (sodium sulfate) and evaporated to dryness. The residue was purified by silica gel chromatography to give V (6.8 g).

Synthesis of intermediate VI: a solution of V (6.0g) in acetic acid (30ml, 80%) was stirred at 80 ℃ for 1 hour. The solvent was removed by evaporation and the residue was purified by silica gel chromatography (DCM/methanol 14: 1) to give VI (3.6 g).

Synthesis of intermediate VII: a solution of VI (3g) and p-toluenesulfonyl chloride (p-TsCl) (3.5g) in pyridine (30ml) was stirred at room temperature for 6 hours. Methanol (5ml) was added and the solvent was evaporated under reduced pressure, the residue was dissolved in EtOAc (3X 150ml) and washed with 0.5M aqueous hydrochloric acid (0 ℃ C.), water (cooled)And a brine (chilled) wash. The combined organic layers were dried (sodium sulfate), filtered through celite and evaporated to dryness. The residue was purified by silica gel chromatography (toluene/ethyl acetate 4: 1) to give VII (3.7 g).

Synthesis of Compound VIII: a solution of VII (3g) and NaN3(2.5g) in DMF (20ml) was stirred at 80 ℃. The reaction mixture was cooled to room temperature and diluted with EtOAc (200mL) and water (50 mL). The organic layer was additionally washed twice with water (2X 50ml) and once with brine (50 ml). All aqueous layers were extracted twice with EtOAc (2X 50 mL). The combined organic layers were dried over sodium sulfate, filtered and the solvent was removed by evaporation. The residue was purified by silica gel chromatography (petroleum ether/EtOAc 5: 2) to give VIII (2.2 g).

Synthesis of Compound X: to a solution of ethyl 2, 3, 4-tri-O-benzyl- α -L-fucothiopyran glycoside IX (1.5g) in DCM (3ml) was added bromine (150 μ L) under argon at 0 ℃. After 5 minutes the cold bath was removed and the reaction mixture was stirred at room temperature for an additional 25 minutes. Cyclohexene (200. mu.l) was added and the reaction mixture was added to VIII (400mg), (Et)₄NBr (750mg) and powderMolecular sieves in DCM (10ml) and DMF (5 ml). After 16 h, triethylamine (1.5ml) was added and stirred for a further 10 min, diluted with EtOAc (50ml) and washed with saturated aqueous sodium bicarbonate solution, water and brine. The aqueous layer was extracted twice with EtOAc ((2X 50 mL.) the combined organic layers were dried (sodium sulfate), filtered and evaporated to dryness and the residue was purified by silica gel chromatography (toluene/EtOAc 9: 1) to give X (700 mg).

Synthesis of Compound XI: to a solution of X (1.5g) in methanol (20ml) was added freshly prepared sodium methoxide (80mg) and the reaction mixture was stirred in a pressure tube at 80 ℃ for 20 hours. The reaction mixture was cooled to room temperature and neutralized with acetic acid. The solvent was evaporated to dryness and the residue was dissolved in diethyl ether. Freshly prepared diazomethane was added and the excess was neutralized with acetic acid. The solvent was removed by evaporation to give XI (1.25 g).

Synthesis of structural unit XV:this synthesis was performed exactly in the same manner as described previously (Helvetica Chimicaacta 83: 2893-2907 (2000)).

Synthesis of compound XVI:XI (1.6g), XV (3g) and activated powdered molecular sieves in DCM (17ml) were stirred at room temperature under argon(1g) For 2 hours. DMTST (2g) was then added in 4 equal portions over a period of 1.5 hours. After 24 h, the reaction mixture was filtered over celite and the filtrate was diluted with DCM (100 ml). The organic layer was washed with saturated aqueous sodium bicarbonate and brine, and the aqueous layer was extracted twice with DCM. The combined organic layers were dried (sodium sulfate), filtered and evaporated to dryness. The residue was purified by silica gel chromatography (toluene/EtOAc 8: 1) to give XVI (1.5 g).

Synthesis of Compound XVII: a solution of triphenylphosphine (500mg in 5ml of dichloromethane) was added dropwise over 10 minutes to a solution of XVI (500mg) and orotyl chloride (500mg) in dichloromethane (10 ml). The reaction mixture was stirred at room temperature for 25 hours and the solvent was removed by evaporation. The residue was purified (silica gel chromatography, DCM/methanol 19: 1) to give XVII (250 mg).

Synthesis of Compound XVIII: to a solution of XVII (200mg) in dioxane-water (5: 1, 12ml) was added 10% Pd-C (100mg) and the reaction mixture was stirred vigorously under hydrogen (55psi) for 24 h. The catalyst was filtered through a layer of celite and the solvent was evaporated off. The residue was purified by silica gel chromatography to give compound XVIII (150 mg).

Synthesis of XIX:to a solution of compound XVIII (145mg) in methanol was added a solution of sodium methoxide in methanol (25%, 0.025ml), and the reaction mixture was stirred at room temperature for 4 hours, neutralized with acetic acid, and the solvent was removed by evaporation. The residue was dissolved in water and passed through a layer of Dowex 50wX-8 (sodium form) resin. Evaporation ofThe aqueous washes were removed to give compound XIX (100 mg).

Synthesis of EDA-XIX: XIX (80mg) and Ethylenediamine (EDA) (1ml) were heated at 70 ℃ and stirred for 5 hours. The solvent was removed by evaporation and purified by Sephadex G-25 column to give EDA-XIX (82 mg).

Example 3: synthesis of Pegylated BASA (FIG. 3)

To a solution of 3, 6-dioxaoctanedioic acid (3, 6-dioxaoctanedioic acid) (PEG, 200mg, commercially available) in DMF (1ml) was added Hunig base (0.4ml), followed by addition of tetramethyluronium Hexafluorophosphate (HATU) (0.35g) after 5 minutes. The solution was stirred at room temperature for 10 minutes, then a solution of BASA (50mg) from example 2 in DMF (0.1ml) was added. The reaction mixture was stirred at room temperature for 4 hours and then the solvent was removed by evaporation. The residue was purified by high pressure liquid chromatography (hplc) (reverse phase C18 column) to give XX (40 mg).

Example 4: glycomimetic Synthesis of BASA Compound #1 (FIG. 4)

To a solution of XX (0.015g) from example 3 in DMF (0.1ml) was added Hunig's base (0.015ml) followed by HATU (0.007 g). The reaction mixture was stirred at room temperature for 10 minutes. A solution of EDA-XIX from example 2 (0.010g in DMF ml) was added and the reaction mixture was stirred at room temperature for 8 hours. The solvent was removed by evaporation and the residue was purified by sephadex G-25 chromatography to give the glycomimetic BASA #1 of FIG. 4 (0.008G).

Example 5: glycomimetic Synthesis of BASA Compound #2 (FIG. 5)

Synthesis of Compound XXI: to a solution of 3, 6-dioxaoctanedioic acid (PEG, 200mg, commercially available) in DMF (1ml) was added Hunig's base (0.4ml), and after 5 minutes HATU (0.35g) was added. The solution was stirred at room temperature for 10 minutes, then a solution of 8-aminonaphthalene-1, 3, 6-trisulfonic acid (50mg, commercially available) in DMF was added. The reaction mixture was stirred at room temperature for 4 hours and the solvent was removed by evaporation. The residue was purified by high pressure liquid chromatography (reverse phase C18 column) to give XXI (25 mg).

Synthesis of Compound XXII: this synthesis was performed in the same manner as described in example 4 except that EDA-XIX and XXI from example 2 were used to give compound XXII (4 mg).

Example 6: effect of Compound #2 on E-selectin and P-selectin AML Rolling on human endothelial cells

The interaction of AML cells with the vascular endothelium is an early stage of extravasation of cancer cells out of the blood circulation. Experiments were performed in the present invention to demonstrate that the human AML cell line (MV-4-11 derived from Biphenotype B myelomonocytic leukemia) interacts with and rolls on Human Umbilical Vein Endothelial Cells (HUVECs) that express E-selectin or P-selectin in vitro. As shown in FIG. 6, Glycomimetic Compound #2 (example 5; FIG. 5) inhibited the rolling of AML cells on stimulated HUVECs.

Example 7: effect of compound #2 on rolling and adhesion of P-selectin mediated multiple myeloma cell line U266 on human endothelial cells under flow conditions.

The interaction of multiple myeloma cells with the vascular endothelium is an early stage of cancer cell extravasation from the bloodstream. Experiments were performed in the present invention to demonstrate that human multiple myeloma cell line (U266) interacts (rolls and adheres) with monolayer Human Umbilical Vein Endothelial Cells (HUVECs) stimulated to express P-selectin. The interaction under normal blood flow shear (1 dyne/cm 2) was quantified by digital image analysis. As shown in FIG. 7, 1.5mM Compound #2 (example 5; FIG. 5) inhibited the interaction of more than 50% of cancer cells with P-selectin expressing monolayers of endothelium.

Example 8: effect of Compound #2 on transmembrane and transendothelial migration of SDF-1 by multiple myeloma cells

In order to home to the bone marrow from the bloodstream or disseminate to other tissues, multiple myeloma cells must first cross the vascular endothelium. An in vitro model of this process was created using a transwell plate, in which each well was divided into 2 chambers by a bisecting membrane. The chemokine, stromal cell derived factor-1 (SDF-1) (30nM), was placed in the lower chamber and the percentage of multiple myeloma cells migrating from the upper chamber to the lower chamber was quantified. Experiments were performed using membranes alone (blank columns) or using membranes covered with a monolayer of endothelial cells (HUVECs). As seen in FIG. 8, Compound #2 (example 5; FIG. 5) specifically inhibited migration through the endothelial monolayer, suggesting that inhibition was based on a target molecule (i.e., selectin) on endothelial cells.

Example 9: effect of Compound #2 on extravasation of multiple myeloma cells from the in vivo blood stream

Human Multiple Myeloma (MM) cells injected into mice spill out of the bloodstream and home to the bone marrow and other organs within minutes. By fluorescently labeling MM cells, this process can be monitored in vivo by biomicroscopy using confocal microscopy. As shown in fig. 9, almost all MM cells had left the peripheral blood stream within 30 minutes. Co-injection of cells with two different doses (100mg/kg and 25mg/kg) of compound #2 significantly inhibited this extravasation of circulating MM cells in vivo.

Example 10: suppression of bone marrow transplantation of AML cells

Acquisition of NOD/SCID/IL2 receptor gamma chain^nullMice ("irradiated mice") (Ishikawa et al, Nat. Biotech.25: 1315-. The AML cell line used in example 6 was intravenously administered to the irradiated mice (control group). The AML cell line was transplanted into the bone marrow of the irradiated mice. Other irradiated mice (experimental group) were administered the Glycomimetic Compound #2 intravenously and then the AML cell line intravenously after a time interval. In another irradiated mouse (different experimental groups), the order of administration was reversed: the AML cell line was first administered intravenously and, after a time interval, glycomimetic compound #2 was administered intravenously. Bone marrow transplantation of AML cell lines was assessed by histological and flow cytometric analysis.

Example 11: effect of Compound #2 on chemotherapy-induced neutropenia

Many chemotherapeutic drugs kill cancer cells by targeting the enhanced cell proliferation associated with malignant tumors. Side effects of these drugs include increased toxicity of normal cells undergoing cell division, such as Hematopoietic Stem Cells (HSCs) required for fresh blood production. In particular, one of the clinically relevant side effects of standard chemotherapeutic drugs in patients is a significant reduction in neutrophils required to fight the infection. Low neutrophil counts contribute to the immune compromised state of cancer patients, which makes them vulnerable to potentially life-threatening infections. The rapid recovery of the immune system after a chemotherapy regimen is a highly desirable goal in this population.

To determine whether selectin inhibition had a beneficial effect on neutrophil protection and recovery from antiproliferative drug treatment, mice were treated with compound #2 before or after 5-fluorouracil (5-FU) or cyclophosphamide administration. At various time points after treatment, blood samples of the mice were taken and analyzed for different cell types including neutrophils.

As illustrated in fig. 10, mice were treated with compound #2 for 14 days by intraperitoneal injection twice a day (50 mg/kg). On day 12 of this treatment period, a cohort (cohort) mice received injections of a chemotherapeutic drug, either 5-fluorouracil (150mg/kg i.p.) or cyclophosphamide (300mg/kg i.p.). On the third day after compound #2 treatment, blood from a cohort of mice was obtained by cardiac puncture to determine the Complete Blood Count (CBC). Other cohorts of mice were bled and analyzed (CBC) on days 5, 7, 9 and 15 after compound #2 treatment.

The chemotherapeutic drugs, 5-FU and cyclophosphamide, have the most significant effect on the number of neutrophils in the blood. Both 5-FU and cyclophosphamide caused severe neutropenia in mice for at least one week. Several days after treatment with these drugs, the number of neutrophils in the blood dropped to dangerously low levels as shown in fig. 11 and 12. Pretreatment of mice with compound #2 significantly improved recovery of neutrophil counts after cyclophosphamide (fig. 11) or 5-FU (fig. 12) administration. Compound #2 treatment itself also promoted an increase in blood levels of neutrophils (data not shown). Although compound #2 did not prevent neutropenia in mice treated with 5-FU or cyclophosphamide, it did promote more rapid and sustained recovery of neutrophils after drug treatment and thus could be combined with standard chemotherapy in cancer patients.

All of the above U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification and/or listed in the application data sheet, are incorporated herein by reference, in their entirety.

From the foregoing, it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention.

Claims (5)

1. Use of a compound, or a physiologically acceptable salt thereof, for the manufacture of a medicament for the treatment of a hematologic cancer or a complication associated therewith, said compound having the formula:

2. the use of claim 1, wherein the physiologically acceptable salt is a sodium salt.

3. The use of claim 1, wherein the complication is metastasis of a hematologic cancer.

4. The use of claim 1, wherein the cancer is acute myeloid leukemia.

5. The use of any one of claims 1-4, wherein the compound or physiologically acceptable salt thereof is in combination with a pharmaceutically acceptable carrier or diluent.