MXPA06005456A - Combinations for the treatment of proliferative diseases - Google Patents

Abstract

The invention features combinations of drugs for the treatment of proliferative diseases (e.g., cancer). The invention also features methods for identifying new combination therapies for the treatment of cancer and other proliferative diseases.

Description

COMBINATIONS FOR THE TREATMENT OF PROLIFERATIVE DISEASES BACKGROUND OF THE INVENTION The present invention relates to the treatment of cancer and other proliferative diseases. Cancer is a disease marked by the uncontrolled growth of abnormal cells. Cancer cells have overcome the barriers imposed on normal cells, which have a finite life time, to grow indefinitely. As the growth of cancer cells continues, genetic alterations may persist until the cancer cell has manifested itself to pursue a more aggressive growth phenotype. If left untreated, metastasis, the expansion of cancer cells to distant areas of the body via the lymphatic system or bloodstream can continue, destroying healthy tissue. Cancer treatment has been hampered by the fact that there is considerable heterogeneity even within a type of cancer. Some cancers, for example, have the ability to invade tissues and display a course of aggressive growth characterized by metastasis. These tumors are usually associated with a poor outcome for the patient. Finally, tumor heterogeneity results in the phenomenon of multiple drug resistance, ie resistance to a broad range of structurally unrelated cytotoxic anti-cancer compounds, JH Gerlach et al., Cancer Surveys, 5: 25-46, 1986. The underlying cause of progressive drug resistance may be due to a small population of drug-resistant cells within the tumor (e.g., mutant cells) at the time of diagnosis, as described, for example, by JH Goldie et al. Andrew J. Coldman, Cancer Research, 44: 3643-3653, 1984. Treating such a tumor with a single drug can result in remission, where the tumor shrinks in size as a result of the death of the predominant drug-responsive cells. However, with the drug-sensitive cells gone, the remaining drug-resistant cells can continue to multiply and eventually dominate the tumor cell population. Therefore, the problems of why metastatic cancers develop pleiotropic resistance to all available therapies, and as this can be counteracted, are the most pressing in cancer chemotherapy. Anti-cancer therapeutic approaches are needed that are reliable for a wide variety of tumor types, and particularly suitable for invasive tumors. Importantly, treatment must be effective with minimal host toxicity. Despite the long history of using multiple combinations of drugs for the treatment of cancer and, in particular, the treatment of cancer resistant to multiple drugs, positive results obtained using combination therapy are still frequently unpredictable. SUMMARY OF THE INVENTION The invention features compositions, methods, and kits for treating proliferative diseases such as cancer. In a first aspect, the invention features a composition that includes a first agent that reduces the biological activity of a mitotic kinesin and a second agent that reduces the biological activity of a protein tyrosine phosphatase. The invention also features a method for treating a patient having a proliferative disease, or inhibiting the development of a proliferative disease in the patient by administering to the patient a first agent that reduces the biological activity of a mitotic kinesin and a second agent that reduces the biological activity of a protein tyrosine phosphatase. Desirably, the two agents are administered simultaneously or within 14 days of each other, within 7 days of each other, within 1 day of each other, within 1 hour of each other in sufficient quantities to treat the patient. The invention also features a method for reducing cell proliferation by contacting a first agent that reduces the biological activity of a mitotic kinesin and a second agent that reduces the biological activity of a protein tyrosine phosphatase. The methods and compositions further include an additional anti-proliferative agent such as an anti-cancer agent. In yet another aspect, the invention presents a method for identifying a combination that may be useful for the treatment of a proliferative disease. In this method, proliferating cells (e.g., cancer cells or a cancer cell line) are contacted in vitro with (i) an agent that reduces the biological activity of mitotic kinesin and (ii) a candidate compound. Using any acceptable assay, it is then determined whether the combination of the agent and the candidate compound reduces cell proliferation, in relation to the proliferation of cells contacted with the agent but not in contact with the candidate compound. A reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease. In another aspect, the invention presents another method for identifying a combination that may be useful for the treatment of a proliferative disease. This method includes the steps of (a) identifying a compound that reduces the biological activity of protein tyrosine phosphatase; (b) contacting proliferative cells in vitro with an agent that reduces the biological activity of mitotic kinesin and the compound identified in step (a); and (c) determining whether the combination of the agent and the compound identified in step (a) reduces cell proliferation, in relation to the proliferation of cells contacted with the agent but not contacted with the compound identified in step (a) or put in contact with the compound identified in step (a) but not in contact with the agent. A reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease. In any of the above aspects, the agent that reduces the biological activity of mitotic kinesin may be, for example, a mitotic kinesin inhibitor, an anti-sense compound or an RNAi compound that reduces the expression levels of a mitotic kinesin, a dominant negative mitotic kinesin, an expression vector encoding such a dominant negative mitotic kinesin, an antibody that binds to mitotic kinesin and reduces the biological activity of mitotic kinesin, or an inhibitor of aurora kinase. Desirably, the agent that reduces the biological activity of mitotic kinesin reduces the biological activity of HsEg5 / KSP. Exemplary mitotic kinesin biological activities are enzymatic activity, motive activity, and binding activity. In still another aspect, the invention presents another method for identifying a compound that may be useful for the treatment of a proliferative disease. This method includes the steps of: (a) providing proliferating cells designed to have reduced mitotic kinesin biological activity; (b) contacting the cells with a candidate compound; Y (c) determining whether the candidate compound reduces cell proliferation, relative to cells not contacted with the candidate compound. A reduction in cell proliferation identifies the compound as a compound that may be useful for the treatment of a proliferative disease. In another aspect, the invention presents yet another method for identifying a combination that may be useful for the treatment of a proliferative disease. This method includes the steps of: (a) contacting proliferating cells in vitro with an agent that reduces the biological activity of protein tyrosine phosphatase and a candidate compound; and (b) determining whether the combination of the agent and the candidate compound reduces cell proliferation, in relation to proliferation of cells contacted with the agent but not in contact with the candidate compound. A reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease. In a related aspect, the invention features a method for identifying a combination that may be useful for the treatment of a proliferative disease. This method includes the steps of: (a) identifying a compound that reduces the biological activity of mitotic kinesin; (b) contacting proliferating cells in vitro with an agent that reduces the biological activity of protein tyrosine phosphatase and the compound identified in step (a); and (c) determining whether the combination of the agent and the compound identified in step (a) reduces the proliferation of cells, in relation to the proliferation of cells contacted with the agent but not contacted with the compound identified in the step (a) or put in contact with the compound identified in step (a) but not put in contact with the agent. A reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease. In any of the above aspects, the agent that reduces the biological activity of protein tyrosine phosphatase is a protein tyrosine phosphatase inhibitor, an antisense compound or an RNAi compound that reduces the expression levels of a protein tyrosine phosphatase, a protein tyrosine phosphatase dominant negative, an expression vector encoding said dominant negative tyrosine phosphatase protein, an antibody that binds to protein tyrosine phosphatase and reduces the biological activity of the protein tyrosine phosphatase, or a farnesyltransferase inhibitor. Desirably, the agent reduces the biological activity of a tyrosine phosphatase protein selected from PTP1B, PRL-1, PRL-2, PRL-3, SHP-1, SHP-2, MKP-1, MKP-2, CDC14, CDC25A , CDC25B, and CDC25C. In another aspect, the invention features another method for identifying a compound that may be useful for the treatment of a proliferative disease. This method includes the steps of: (a) providing proliferating cells designed to have reduced protein tyrosine phosphatase biological activity; (b) contacting the cells with a candidate compound; and (c) determining whether the candidate compound reduces cell proliferation, relative to cells not contacted with the candidate compound. A reduction in cell proliferation identifies the compound as a compound that may be useful for the treatment of a proliferative disease. In any of the above aspects, the cells are preferably cancer cells or cells from a cancer cell line. By "more effective" it is understood that a method, composition, or kit exhibits greater efficacy, is less toxic, safer, more convenient, better tolerated, or less expensive, or provides more treatment satisfaction than another method, composition or kit with the one that is being compared. The efficiency can be measured by a person skilled in the art using any standard method that is appropriate for a given indication. By "mitotic kinesin inhibitor" is meant an agent that binds to a mitotic kinesin and reduces, by a significant amount (e.g., at least 10%, 20%, 30% or more), the biological activity of that Kinesis kinesin. The biological activities of mitotic kinesin include enzymatic activity (e.g., ATPase activity), motor activity (e.g., force generation) and linkage activity (e.g., linking of the motor to any of its micro -Tubles or their load). By "dominant negative" is meant a protein that contains at least one mutation that deactivates its physiological activity such that the expression of this mutant in the presence of the normal or wild type copy of the protein results in deactivation or reduction of the activity of the normal copy. Thus, the activity of the mutant "dominates" over the activity of the normal copy such that although the normal copy is present, the biological function is reduced. In one example, a dimer of two copies of the protein is required such that even if a normal copy and a mutated copy are present there is no activity; another example is when the mutant is linked to or "soaks" other proteins that are critical for normal copy function such that not enough of these other proteins are present for normal copy activity. By "protein tyrosine phosphatase" or "ATPase" is meant an enzyme that dephosphorylates a tyrosine residue on a protein substrate. By "protein tyrosine phosphatase inhibitor" is an agent that binds to protein tyrosine phosphatase and inhibits (eg., by at least 10%, 20%, or 30% or more) the biological activity of that protein tyrosine phosphatase. By "dual specification phosphatase" is meant a phosphatase protein that can dephosphorylate both a tyrosine residue and a residue of either serine or threonine in the same protein substrate. The dual-specification phosphatases include MKP-1, MKP-2, and the cell division cycle phosphatase family (e.g., CDC14, CDC25A, CDC25B, and CDC25C). The phosphatases of dual specification are considered to be tyrosine phosphatase proteins. By "anti-proliferative agent" is meant a compound that, individually, inhibits cell proliferation. The anti-proliferative agents of the invention include alkylating agents, platinum agents, anti-metabolites, topoisomerase inhibitors, anti-tumor antibiotics, anti-mitotic agents, aromatase inhibitors, thymidylate synthase inhibitors, DNA antagonists, farnes inhibitors. -tiltransferase, pump inhibitors, histone acetyltransferase inhibitors, metalloproteinase inhibitors, ribunucleoside reductase inhibitors, TNF alpha agonists and antagonists, endothelin A receptor antagonists, retinoic acid receptor agonists, immuno-modulators, hormonal and anti-inflammatory agents -hormonal, photodynamic agents, and tyrosine kinase inhibitors. By "inhibiting cell proliferation" is meant that it reduces, stops, or measurably reverses the cell growth rate in vitro or in vivo. Desirably, a reduction of the growth rate is by at least 20%, 30%, 50%, 60%, 70%, 80%, or 90%, as determined using a suitable assay for determination of the growth rates of cells (e.g., a cell growth assay described herein). Typically, an inversion of the growth rate is achieved by initiating or accelerating the necrotic or apoptotic mechanisms of cell death in neoplastic cells. By "a sufficient amount" is meant the amount of a compound, in a combination according to the invention, required to inhibit the growth of the cells of a neoplasm in vivo. The effective amount of active compounds used to practice the present invention for the treatment of proliferative diseases (ie, cancer) varies depending on the manner of administration, age, race, sex, affected organ, body weight, and general health of the subject . Finally, the attending doctor or veterinarian will decide the appropriate amount and dosage regimen. A "low dose" means at least 5% less (eg, at least 10%, 20%, 50%, 80%, 90%, or even 95%) than the lowest recommended standard dose of a particular compound formulated for a given administration route for treatment of any human disease or condition. A "high dose" means at least 5% (eg, at least 10%, 20%, 50%, 100%, 200%, or even 300%) more than the highest recommended standard dose of a particular compound for treatment of any disease or human condition. The phrase "pharmaceutically acceptable" refers to "Molecular identities and compositions that do not produce an adverse, allergic, or other non-directed reaction when administered to the patient. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and delaying absorption agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. By "patient" is meant any animal (eg, a human). Non-human animals that can be treated using the methods, compositions, and kits of the invention include horses, dogs, cats, pigs, goats, rabbits, hamsters, monkeys, piglets, rats, mice, lizards, snakes, sheep, cows , fish, and birds. Compounds useful in the invention include those described herein in any of their pharmaceutically acceptable forms, including isomers such as diastereomers and enantiomers, salts, solvates, and polymorphs thereof, as well as racemic mixtures of the compounds described herein. Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

Detailed Description The invention features compositions, methods, and kits for treating proliferative disorders. Normal cells have signaling mechanisms that regulate growth, mitosis, differentiation, cell function, and death in a programmed manner. Defects in the signaling pathways that regulate these functions can result in uncontrolled growth and proliferation, which can manifest as cancer, hyperplasia, restenosis, cardiac hypertrophy, immune disorders and inflammatory disorders. The mitotic kinesins are essential engines in mitosis. They control the assembly and maintenance of spindles (bundles of micro-tubules), union and proper positioning of the chromosomes in the spindle, establish the bipolar spindle and maintain forces in the spindle to allow the movement of chromosomes towards opposite poles. Perturbations of the mitotic kinesin function cause malformation or dysfunction of the mitotic spindle, frequently resulting in cell cycle arrest and cell death. The protein tyrosine phosphatases (ATPases) are intracellular signaling molecules that dephosphorylate a tyrosine residue in a protein substrate, thereby modulating certain cellular functions. In normal cells, they typically act in conjunction with protein tyrosine kinases to regulate signaling cascades through the phosphorylation of tyrosine protein residues. The phosphorylation and dephosphorylation of tyrosine residues in proteins controls the growth and proliferation of cells, cell cycle progression, cytoskeletal integrity, differentiation and metabolism. In several metastatic and cancer cell lines, PTP1B and the Regenerating Liver Phosphatase family (PRL-1, PRL-2, and PRL-3) have been shown to be over-expressed. For example, PRL-3 (also known as PTP4A3) is expressed at relatively high levels in metastatic colorectal cancers (Saha et al., Science 294: 1343-1346, 2001). PRL-1 localizes to the mitotic spindle and is required for mitotic progression and segregation of chromosomes. PRL phosphatases promote cell migration, invasion, and metastasis, and the inhibition of these ATPases has been shown to inhibit the proliferation of in vitro cancer cells and tumors in animal models. It was previously shown that the combination of chlorpro-mazine and pentamidine work together to reduce cell proliferation (US Pat. No. 6,569,853). Now it is shown that chlorpromazine acts as an inhibitor of mitotic kinesin. Pentamidine has also been shown to be an inhibitor of PRL phosphatases (Pathak et al., Mol. Cancer Ther., 1: 1255-1264, 2002). Based on the above observations, it is concluded that combinations of an agent that reduce the biological activity of a mitotic kinesin with an agent that reduces the activity of a protein tyrosine phosphatase are useful for reducing cell proliferation and, therefore, for treat proliferative diseases. Kinesic kinesins Kinesis kinesins include HsEg5 / KSP, KIFC3, CH02, MKLP, MCAK, Kin2, Kif4, MPPl, CENP-3, NYREN62, LOC8464, and KIF8. Other mitotic kinesins are described in patents US 6,414,121, 6,582,958, 6,544,766, 6,492,158, 6,455,293, 6,440,731, 6,437,115, 6,420,162, 6,399,346, 6,395,540, 6,383,796, 6,379,941, and 6,248,594. The GenBank accession numbers of representative mitotic kinesins are given in Table 1.

HsEg5 / KSP has been cloned and characterized (see, e.g.

Blangy et al., Cell, 83: 1159-69 (1995); Galgio et al., J. Cell Biol., 135: 399-414,1996; Whitehead et al., J. Cell Sci., 111: 2551-2561, 1998; Kaiser et al., J. Biol. Chem., 274: 18925-31, 1999; GenBank access numbers: X85137, NM 004523). Homologs of KSP Drosophi-la (Heck et al., J. Cell Biol., 123: 665-79, 1993) and Xenopus (Le Guellec et al., Mol Cell Biol., 11: 3395-8, 1991) have been reported. . Drosophila KLP61F / KRP130 has been reportedly purified in a native manner (Colé et al., J. Biol. Chem., 269: 22913-22916, 1994), expressed in E. coli, (Barton et al., Mol. Biol. Cell, 6: 1563-74,1995) and it has been reported to have motility and ATPase activities (Colé et al, supra, Barton et al., Supra). Xenopus Eg5 / KSP was expressed in E. coli and was reported to possess motility activity (Sawin et al., Nature, 359: 540-3, 1992; Lockhart and Cross, Biochemistry, 35: 2365-73, 1996; Crevel et al., J. Mol. Biol., 273: 160-170, 1997) and activity of ATPase (Lockhart and Cross, supra, Crevel et al., Supra). In addition to KSP, other members of the BimC family include BimC, CIN8, cut7, KIP1, KLP61F (Barton et al., Mol. Biol. Cell 6: 1563-1574, 1995; Cottingham &Hoyt, J. Cell Biol. : 1041-1053, 1997; DeZwaan et al., J. Cell Biol. 138: 1023-1040, 1997; Gaglio et al., J. Cell Biol. 135: 399-414, 1996; Geiser et al., Mol. Biol. 8: 1035-1050, 1997; Heck et al., J. Cell Biol. 123: 665-679, 1993; Hoyt et al., J. Cell Biol. 118: 109-120, 1992; Hoyt et al., Genetics 135: 35-44, 1993; Huyett et al., J. Cell Sci. 111: 295-301, 1998; Miller et al., Mol. Biol. Cell 9: 2051-2068, 1998; Roof et al., J. Cell Biol. 118: 95-108, 1992; Sanders et al., J. Cell Biol. 137: 417-431, 1997; Sanders et al., Mol. Biol. Cell 8: 1025-0133, 1997; Sanders et al., J. Cell Biol. 128: 617-624, 1995; Sanders & Hoyt, Cell 70: 451-458, 1992; Sharp et al., J. Cell Biol. 144: 125-138, 1999; Straight et al., J. Cell Biol. 143: 687-694, 1998; Whitehead & Rattner, J. Cell Sci. 111: 2551-2561, 1998; Wilson et al., J. Cell Sci. 110: 451-464, 1997). The biological activities of the mitotic kinesin include its ability to affect the hydrolysis of TPA; linked micro-tubules; slip and polymerization / depolymerization (effects on the dynamics of micro-tubules); linked to other spindle proteins; linked to proteins involved in the control of the cell cycle; serve as a substrate to other enzymes, such as kinases or proteases; and specific kinesin cell activities such as spindle pole separation. Methods for assaying biological activity of a mitotic kinesin are well known in the art. For example, methods for carrying out motility tests are described, e.g., in Hall et al., 1996, Biophys. J., 71: 3467-3476; Turner et al., 1996, Anal. Biochem. 242: 20-25; Gittes et al., 1996, Biophys. J. 70: 418-429; Shirakawa et al., 1995, J. Exp. Biol. 198: 1809-1815; Winkelmann et al., 1995, Biophys. J. 68: 2444-2453; and Winkelmann et al., 1995, Biophys. J. 68: 72S. Methods known in the art for determining ATPase hydrolysis activity can also be used. The patent application US 09 / 314,464, filed on May 18, 1999, incorporated herein by reference in its entirety, describes such tests. Other methods can also be used. For example, release of Pi from kinesin can be quantified. In one embodiment, the assay of ATP hydrolysis activity utilizes 0.3 M perchloric acid (PCA) and malachite green reagent (8.27 mM sodium molybdate II, 0.33 mM malachite oxalate 0.33 mM, and 0.8 mM Triton X-100. ). To carry out the assay, 10 μL of reaction are depleted in 90 μL of cold 0.3 M PCA. Phosphate standards are used such that data can be converted to nM inorganic phosphate released. When all reactions and standards have been depleted in PCA, 100 μL of malachite green reagent is added to relevant wells in, eg, a micro-titration plate. The mixture is developed for 10-15 minutes and the plate is read at an absorbance of 650 nm. If phosphate standards are used, abosrbancy readings can be converted to nM of P ± and plotted against time. Additionally, ATPase assays known in the art include the luciferase assay.

The ATPase activity of kinesin motor domains can also be used to monitor the effects of modulating agents. In one embodiment kinase assays of ATPase are carried out in the absence of micro-tubules. In another embodiment, the ATPase assays are carried out in the presence of micro-tubules. Different types of modulating agents can be detected in the previous tests. In an embodiment, the effect of a modulating agent is independent of the concentration of micro-tubules and ATP. In another embodiment, the effect of the agents on the kinesin ATPase can be decreased by increasing the concentrations of ATP, micro-tubules, or both. In yet another embodiment, the effect of the modulating agent is increased by increasing the concentrations of ATP, micro-tubules, or both. Agents that reduce the biological activity of an in vitro mitotic kinesin can be selected in vivo. Methods for in vivo selection include assays for cell cycle distribution, cell viability, or the presence, morphology, activity, distribution, or number of mitotic spindles. Methods for monitoring the cell cycle distribution of a population of cells, for example, by flow cytometry, are well known to those skilled in the art, as are methods for determining cell viability (see, e.g., US 6,617,115). Inhibitors of mitotic kinesin The inhibitors of mitotic kinesin include chlorpromazine, monasterol, terpendola E, HR22C16, and SB715992. Other inhibitors of mitotic kinesin are those compounds disclosed in Hopkins et al., Biochemistry 39: 2805, 2000; Hotha and collaborators, Angew. Chem. Inst. Ed. 42: 2379, 2003; PCT publications WO 01/98278, WO 02/057244, WO 02/079169, WO 02/057244, WO 02/056880, WO 03/050122, WO 03/050064, WO 03/049679, WO 03/049678, WO 03 / 049527, WO 03/079973, and WO 03/039460, and the patent application publications US 2002/0165240, 2003/0008888, 2003/0127621, and 2002/0143026; and US patents, 6,437,115, 6,545,004, 6,562,831, 6,569,853, and 6,630,479, and the chlorpromazine analogues described in US patent application 10 / 617,424 (see, eg, the formula (D) Protein shots ± na phosphatases Proteins tyrosine phosphatases include the PRL family (PRL-1, PRL-2, and PRL-3), PTP1B, SHP-1, SHP-2, MKP-1, MKP-2, CDC14, CDC25A, CDC25B, CDC25C, PTPa, and PTP-BL. The biological activities of the tyrosine phosphatases proteins include the dephosphorylation of tyrosine residues in substrates. GenBank accession numbers of representative tyrosine phosphatases are provided in Table 2.

Tyrosine phosphatase protein inhibitors Protein tyrosine phosphatase inhibitors include pentamidine, levamisole, ketoconazole, bisperoxane-nadium compounds (e.g., those described in Scrivens et al., Mol. Cancer Ther. 2: 1053-1059, 2003, and US Pat. No. 6,642,221), vanadate salts and complexes (e.g., sodium orthovanadate), desfosfatin, dnacin Al, dnacin A2, STI-571, suramin, gallium nitrate, sodium stibogluconate, meglumine antimonate, 2- (2-mercaptoethanol) -3-methyl-1,4-naphthoquinone, 2,5-bis (4-amidinophenyl) furan-bis-O-methylamidoxime, known as DB289 (Immtech), 2,5-bis (4-amidinophenyl) ) furan (DB75, Immtech), disclosed in US patent 5,843,980, and compounds described in Pestell et al., Oncogene 19: 6607-6612, 2000; Lyon et al., Nat. Rev. Drug Discov. 1: 961-976, 2002; Ducruet et al., Bioorg. Med. Chem. 8: 1451-1466, 2000, and US 2003/0114703, 2003/0144338, and 2003/0161893 patent application publications, and PCT patent publications WO 99/46237, WO 03/06788 and WO 03/070158. Still other analogs are those that fall within a formula provided in any of the patents US 5,428,051; 5,521,189; 5,602,172; 5,643,935; 5,723,495; 5,843,980; 6,008,247; 6,025,398; 6,172,104; 6,214,883; and 6,326,395, and the patent application publications US 2001/0044468 and 2002/0019437, and the pentamidine analogs described in the patent application US 10 / 617,424 (see, e.g., Formula (II)). Other protein tyrosine phosphatase inhibitors can be identified, for example, using the methods described in Lazo et al. (Oncol. Res. 13: 347-352, 2003), PCT publications WO 97/40379, WO 03/003001, and WO 03/035621, and US Patents 5,443,962 and 5,958,719. Other inhibitors of biological activity In addition to reducing biological activity through the use of compounds that bind to a mitotic kinesin or a protein tyrosine phosphatase, other inhibitors of the biological activity of mitotic kinesin and protein tyrosine phosphatase can be employed. Such inhibitors include compounds that reduce the amount of target protein or RNA levels (e.g., anti-sense compounds, dsRNA, ribosomes) and compounds that compete with mitotic kinesins or endogenous tyrosine phosphatase proteins to bind partners (e.g. , dominant negative proteins or polynucleotides that encode them). Anti-sense compounds The biological activity of a mitotic kinesin and / or a protein tyrosine phosphatase can be reduced through the use of an anti-sense compound directed to RNA encoding the target protein. Anti-sense kinesin kinesin compounds for this use are known in the art (see, e.g., US Pat. No. 6,472,521, WO 03/030832, and Maney et al., J. Cell Biol., 1998, 142: 787- 801), as are anti-sense compounds against tyrosine phosphatase proteins (see, e.g., US Patent Publication 2003/0083285 and Weil et al., Biotechniques 33: 1244, 2002). Other anti-sense compounds that reduce mitotic kinesins can be identified using standard techniques. For example, accessible regions of the mitotic kinesin mRNA or target protein tyrosine phosphatase can be predicted using a secondary RNA structure development program such as MFOLD (M. Zuker, DH Mathe s &DH Turner, Algorithms and Thermomics for RNA Secondary Structure Prediction: Practical Guide In: RNA Biochemistry and Biotechno-logy, J. Barcisze ski &BFC Clark, editors, NATO ASI Series, Kluwer Academic Publishers, (1999)). Sub-optimal developments with a free energy value within 5% of the most stable predicted envelopment of the mRNA are predicted using a 200 base window within which a residue can find a complementary base to form a base pair link. Open regions that do not form a base pair are added together with each sub-optimal envelopment and areas that are predicted to be open are considered more accessible when linked to core-base antisense oligomers. Other methods for anti-sense design are described, for example, in US Pat. No. 6,472,521, Antisense Nucleic Acid Drug Dev. 1997, 7: 439-444, Nucleic Acids Research 28: 2597-2604, 2000, and Nucleic Acids Research 31: 4989-4994, 2003. RNA interference The biological activity of a mitotic kinesin and / or protein tyrosine phosphatase can be reduced through the use of RNA interference (RNAi), employing, e.g., a double-stranded RNA (dsRNA). ) or small interfering RNA (siRNA) targeted to the mitotic kinesin or protein tyrosine phosphatase in question (see, e.g., Miyamoto et al., Prog. Cell Cycle Res. 5: 349-360, 2003).; US 2003/0157030 patent application publication). Methods for designing such interference RNAs are known in the art. For example, software to design RNA interference is available from Oligoengine (Seattle, Washington, United States).

Dominant Negative Proteins A person skilled in the art will know how to make mitotic kinesins and dominant tyrosine phosphatase proteins. Such dominant negative proteins are described, for example, in Gupta et al., J. Exp. Med., 186: 473-478, 1997; Maegawa et al., J. Biol. Chem. 274: 30236-30243, 1999; Woodford-Thomas et al., J. Cell Biol. 117: 401-414, 1992. Aurora kinase inhibitors Aurora kinases have been shown to be protein kinases of a new family that regulate the structure and function of the mitotic spindle. One target of aurora kinases include mitotic kinesins. The aurora kinase inhibitors can thus be used in combination with a compound that reduces the biological activity of protein tyrosine phosphatase according to a method, composition, or kit of the invention. There are three classes of aurora kinases: aurora-A, aurora-B and aurora-C. Auroras-A includes AIRK1, DmAurora, HsAurora-2, HsAIK, HsSTKld, CeAIR-1, MmARKl, MmAYKl, MmlAKl and XIEg2. Auroras-B include AIRK-2, DmIAL-1, HsAurora-1, HsAIK2, HsAIM-1, HsSTK12, CeAIR-2, MmARK2 and XAIRK2. Auroras-C include HsAIK3 (Ada s et al., Trends Cell Biol. 11: 49-54, 2001). Aurora kinase inhibitors include VX-528 and ZM447439; others are described, e.g., in the patent application publication 2003/0105090 and in patents US 6,610,677, 6,593,357, and 6,528, 509.

Farnesyltransferase Inhibitors Farnesyltransferase inhibitors alter the biological activity of PRL phosphatases and thus may be used in combination with a compound that reduces the activity of mitotic kinesin in a method, composition, or kit of the invention. Inhibitors of farnesyltransferase include arglabine, lonafar-nib, BAY-43-9006, tipifarnib, perilylic alcohol, FTI-277 and BMS-214662, as well as those compounds described, e.g., in Kohl, Ann. NY Acad. Sci. 886: 91-102, 1999, the publications of patent applications US 2003/0199544, 2003/0199542, 2003/0087940, 2002/0086884, 2002/0049327, and 2002/0019527, US patents 6,586,461 and 6,500,841, and WO 03/004489. Therapy The compounds of the invention are useful for the treatment of cancers and other disorders characterized by hyperproliferative cells. The therapy can be carried out alone or in conjunction with another therapy (e.g., surgery, radiation therapy, chemotherapy, immunotherapy, anti-angiogenesis therapy, or gene therapy). Additionally, a person having a higher risk of developing a neoplasm or other proliferative disease (e.g., one who is genetically predisposed or one who has previously had such a disorder) may receive prophylactic treatment to inhibit or delay hyperproliferation. The duration of the combination therapy depends on the type of disease or disorder being treated, the age and condition of the patient, the stage and type of the paicente's disease, and how the patient responds to treatment. Therapy can occur in active and inactive cycles that include rest periods such that the patient's body has an opportunity to recover from any unforeseen side effects. Desirably, the methods, compositions, and kits of the invention are more effective than other methods, compositions, and kits. By "more effective" is meant that a method, composition or kit exhibits greater efficacy, is less toxic, safer, more convenient, better tolerated, or less expensive, or provides greater treatment satisfaction than another method, composition, or kit with which is being compared. Cancers include, without limitation, leukemias (e.g., acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (Hodgkin's disease, not Hodgkin's disease), Waldenstrom's macroglobuli-nemia, heavy chain disease, and solid tumors such as sarcomas and carcinomas (eg, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, sinovioma, mesothelioma) , Ewing tumor, leyomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronguiogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, veji carcinoma ga, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, meningioma, melanoma, neuroblastoma, and retinoblastoma). Other proliferative diseases that can be treated with the combinations and methods of the invention include lymphoproliferative disorders and psoriasis. By "lymphoproliferative disorder" is meant a disorder in which there is an abnormal proliferation of cells of the lymphatic system (e.g., T cells and B cells). Additionally therapy may include the use of other anti-proliferative agents with the combinations of the invention. For example, when treatment is for cancer, the combination can be administered with an anti-cancer agent, such as the agents in Table 3, below.

Table 3 Busulfan procarbazine dacarbazine altretamine ifosfamide estramustine phosphate hexamethyl elastin mechlorethamine Alkylating agents thiotepa streptozocin dacarbazine temozolomide lomustine semustine cyclophosphamide cisplatin chlorambucil Spiroplatin lobaplatin (Aeterna) tetraplatin satraplatin (Johnson Matthey) ormaplatin BBR-3464. (Hoffman-La Roche) Platinum agents iproplatin SM-11355 (Sumitomo) ZD-0473 (AnorMED) AP-5280 (Access) Carboplatin oxaliplatin azacitidine tri etrexate floxuridine deoxicoformycin 2-chlorodeoxyazidosine pentostatin 6-mercaptopurine hydroxyurea 6-thioguanine decitabine (SuperGen) Anti- etabolites cytarabine clofarabine (Bioenvision) 2-fluorodeoxycytidine irofulvenene (MGI Pharma) methotrexate DMDC (Hoffmann- a Roche) to udex ethinylcytidine (Taiho) fludarabine gemcitabine raltitrexed capecitabine amsacrine exatecan mesylate (Daiichi) quinamed epirubicin (ChemGenex) etoposide gimatecan (Sigma-Tau) teniposide or mitoxantrone diflomotecan (Beaufour-Ipsen) 7-ethyl-10-hydroxy-captothecin TAS-103 (Taiho) dexrazoxanet (TopoTarget) elsamitrucine (Spectrum ) Pixantrone Inhibitors (Novusp Weapon) J-107088 (Merck &Co) rebeccamycin analog topoisomerase BNP-1350 (BioNumerik) (Exelixis) CKD-602 (Chong Kun Dang) BBR-3576 (Novuspharma) K -2170 (Kyowa Hakko) rubitecan (SuperGen) hydroxycaptothecin (SN-38) irinotecan (CPT-11) topotecan dactinomycin (actinomycin D) azonafide 1 valrubicin anthrapirazole daunorubic na (daunomycin) oxantrazole terarubicin losoxantrone Antibiotics idarubxcin bleomycin acid anti-tumor rubidazone MEN-10755 (Menarini) plicamycin GPX -100 (Gem Pharmaceuticals) porfiromycin epirubicin itoxantrone (novantrone) itoxantrone amonafide colchicine E7010 (Abbott) vinblastine PG-TXL (Cell Therapeutics) vindesine IDN 5109 (Bayer) dolastatin 10 (NCI) A 105972 (Abbott) rhizoxin (Fujisawa) A 204197 (Abbott) my obulin (Warner-Lambert) Lü 223651 (BASF) cemadotine (BASF) D 24851 (ASTAMedica) RPR 109881A (Aventis) ER-86526 (Eisai) TXD 258 (Aventis) combretastatin A4 (BMS) epothilone B (Novartis) isohomohalicondrine-B (PharmaMar) Agents T 900607 (Tularik) ZD 6126 (AstraZeneca) anti-mitotic T 138067 (Tularik) AZ 10992 (Asahi) cryptophycin 52 (Eli Lilly) IDN-5109 (Indena) vinflunine (Fabre) AVLB (Prescient NeuroPharma) auristatin PE (Teikoku Hormone ) azaepotilone B (BMS) BMS 247550 (BMS) BNP-7787 (BioNumerik) BMS 184476 (BMS) CA-4 prodrug (OXiGENE) BMS 188797 (BMS) dolastatin-10 (NIH) taxoprexin (Protarga) CA-4 (OXiGENE) SB 408075 (GlaxoSmith line) docetaxel vinorelbine vincristine paclitaxel aminoglutethimide YM-511 (Yamanouchi) Atamestane inhibitors (BioMedicines) formestane aromatase letrozole exe stannas anastrazole Inhibitors of pemetrexed (Eli Lilly) nolatrexed (Eximias) thymidylate synthase ZD-9331 (BTG) CoFactor (BioKeys) trabectedin (PharmaMar) edotreotide (Novartis) glufosfamide (Baxter mafosfa ida (Baxter International) International) DNA albumin + 32P antagonists (Isotope apaziquona (Spectrum Solutions) Pharmaceuticals) txiriectacina (New Biotics) 06 bencil guanina (Paligent) arglabina (NuOncology Labs) tipifarnib (Johnson s Johnson) Inhibitors of lonafarnib (Schering-Plow) perilylic alcohol (DOR farnesyltransferase BAY-43-9006 (Bayer) BioPharma) CBT-1 (CBA Pharma) zosuquidar trihydrochloride Inhibitors of tariquidar (Xenova) (Eli Lilly) pump MS-209 (Schering AG) dicirrato de biricodar (Vértex) Inhibitors of tacedinalin (Pfizer) pivaloyloxymethyl butyrate histone SAHA (Aton Pharma) (Titan) acetyltransferase M? -275 (Schering AG) depsipeptide (Fujisawa) Inhibitors of Neovastat (Aeterna Laboratories) CMT-3 (CollaGenex) metalloproteinase marimastat (British Biotech) BM? -275291 (Celltech) Inhibitors of gallium ribo- maltolate (Titan) tezacitabine (Aventis) nucleoside reductase triapine (Vion) didox (Molecules for Health) Virulizine agonists / antagonists (Lorus Therapeutics) revimid (Celgene) of TNF alpha CDC-394 (Celgene) Antagonist of the atrasentan receptor (Abbott) YM-598 (Yamanouchi) of Endothelium A ZD-4054 (AstraZeneca) Receptor agonists fenretinide (Johnson s Johnson) alitret noina (Ligand) retinoic acid LGD-1550 (Ligand) interferon dexosome therapy (Anosys) oncophagus (Antigenics) pentrix (Australian Cancer GMK (Progenies) Technology) adenocarcinoma vaccine ISF-154 (Tragen) (Biomira) cancer vaccine (Intercell) Immuno-modulators CTP-37 (AVI BioPharma) norelin (Biostar) IRX-2 (Immuno-Rx) BLP-25 (Biomira) PEP-005 (Peplin Biotech) MGV (Progenies) vaccines of sincrovax (CTL Immuno) ß-aletine ( Dovetail) melanoma vaccine (CTL Immuno) CLL therapy (Vasogen) p21 RAS vaccine (GemVax) estrogen dexamethasone conjugated estrogens prednisone ethinyl estradiol methylprednisone clortiranisine prednisolone idenestrol aminoglutethimide hydroxyprogesterone caproate leuprolide medroxyprogesterone octreot da Hormone testosterone mitotane and anti-hormonal propionate of testosterone P-04 (Novogen) fluoxymesterone 2-methoxyestradiol (EntreMed) methyltestosterone arzoxifen (Eli Lilly) diethylstilbestrol tamoxifen megestrol toremofine bicalutamide goserelin flutamide leuporelin nilutamide bicalutamide Talaporfin (Light Sciences) Pd-baceteriofeoforbida (Jedda) Theralux (Theratechnologies) texaphyrin lutetium Photodynamic agents motexafin gadolinium (Pharmacyclics) (Pharmacyclics) hypericin imatinib (Novartis) EKB-569 (Wyeth) leflunomide (Sugen / Pharmacia) kahalide F (PharmaMar) ZD1839 (AstraZeneca) CEP-701 (Cephalon) erlotinib (Oncogene Science) CEP-751 (Cephalon) canertinib (Pfizer) MLN518 (Millenium) escuala ina (Genaera) PKC412 (Novartis) SU5416 (Pharmacia) Phenoxodiol (Novogen) S06668 (Pharmacia) C225 (ImClone) Kinase inhibitors ZD4190 (AstraZeneca) rhu-Mab (Genetech) ZD6474 (AstraZeneca) MDX-H210 (Medarex) vatalanib ( Novartis) 2C4 (Genetech) PK1166 (Novartis) MDX-447 (Medarex) GW2016 (GlaxoSmith line) ABX-EGF (Abgenix) EKB-509 (Wyeth) IMC-1C11 (I Clone) trastuzu ab (Genetech) Tyrphostins Gefitinib (Iressa) Miscellaneous agents SR-27897 (inhibitor of CCK A, Sanofi ceflatonin (promoter of apoptosis, ChemGenex) Synthelabo) BCX-1777 (PNP inhibitor, BioCryst) tocladesine (cyclic AMP agonist, ranpirnase (ribonuclease stimulant, Ribapharm) Alfacell) alvocidib (inhibitor of CDK, Aventis) galarubicin (inhibitor of RNA synthesis, CV-247 (COX-2 inhibitor, Ivy Medical) Dong-A) P5 (COX-2 inhibitor, Phytopharm) tirapazamine (reducing agent, SRI CapCell (stimulant of CYP450, Bavarian International) Nordic) N-acetylcysteine (reducing agent) , Zambón) GCS-100 (Gal3 antagonist, Glycogenesis) R-flurbiprofen (inhibitor of NF-kappaB, immunogen G17DT (gastrin inhibitor, Encoré) Aphton) 3CPA (inhibitor of NF-kappaB, Active Biotech) efaproxiral (oxygenator, Allos Therapeutics) seocalcitol (receptor agonist of PI-88 (heparanase inhibitor, Progen) vitamin D, Leo) tesmilfen (histamine antagonist, YM 131-I-TM-601 (DNA antagonist, Bio? ciences) TransMolecular) histamine (receptor agonist) of histamine eflornithine (ODC inhibitor, ILEX Oncology) H2, Maxim) inodronic acid (osteoclast inhibitor, thiazofurin (inhibitor of IMPDH, Ribapharm) Yamanouchi) cilengitide (integrin antagonist, Merck indisulam (stimulant of p53, Eisai) K Ga A) aplidine (inhibitor of PPT, PharmaMar) SR-31747 (IL-1 antagonist, Sanofi-gemtuzumab (CD33 antibody, Wyeth Ayerst) Synthelabo) PG2 (hematopoiesis enhancer, CCI-779 (mTOR kinase inhibitor, Wyeth) Pharmagenesis) exisulind (PDE V inhibitor, Cell Pathways) Immunol (triclosan mouthwash, Endo) CP-461 (PDE inhibitor V, Cell Pathways) triacetyluridine (prodrug of uridine, AG-2037 (GART inhibitor, Pfizer) Wellstat) WX-üKl (SN-4071 activator inhibitor (sarcoma agent, Signature plasminogen, Wilex) Bio? Cience) PBI-1402 (PMN stimulant, ProMetic TransMID-107 ( immunotoxin, KS Biomedix) LifeSciences) PCK-3145 (promoter of apoptosis, Procyon) bortezomib (proteasome inhibitor, doranidazole (promoter of apoptosis, Pola) Millenium) CHS-828 (cytotoxic agent, Leo) SRL-172 (T-cell stimulant, SR Pharma) trans-retinoic acid (differentiator, NIH) TLK-286 (glutathione inhibitor S MX6 (promoter of apoptosis, MAXIA) transferase, Telik) apo ina (promoter of apoptosis, ILEX Oncology) PT-100 (agonist of the growth factor, uroc dina (promoter of apoptosis, Bioniche) Point Therapeutics) Ro-31-7453 (promoter of apoptosis, La Roche) midostaurin (inhibitor of PKC, Novartis) brostalicin (apoptosis promoter, briostatin-1 (PKC stimulant, GPC Pharmacia) Biotech) CDA-II (apoptosis promoter , Everl fe) SDX-101 (promoter of apoptosis, Salmedix) rituximab (CD20 antibody, Genentech) Examples The following examples are to illustrate the invention. They are not intended to limit the invention in any way. Chlorpromazine is a mitotic kinesin inhibitor. Chlorpromazine was determined to be a mitotic kinesin inhibitor using a free cell engine assay. This assay measures the organic phosphate (Pi) generated during ATPase activity activated by micro-tubules of kinesin engine proteins. Recombinant kinesin motor protein activity of HsEg5 / KSP was assayed using the Kinesin ATPase Kinesin Endpoint Biochem Kit (Cytoskeleton, catalog number BK053) followed by the manufacturer's instructions for amounts of reaction buffer, ATP and micro-tubules. The amount of kinesin protein HsEg5 / KSP was optimized to 0.8 μg per reaction and included where indicated. Each assay was carried out in a total reaction volume of 30 μL in a clear 96-well area plate (Corning Inc., Costar and catalog number 3697) and included the following conditions: 1. a reaction control consisting of reaction buffer and ATP only; 2. Negative control reactions containing: a. micro-tubules and ATP without kinesin protein or b. kinesin HsEg5 / KSP and ATP without micro-tubules; Y 3. experimental reactions containing ATP, kinesin, and micro-tubules with or without compound in the final concentrations indicated. The reactions were pre-incubated for 15 minutes at room temperature prior to the addition of ATP. After addition of ATP, -the reaction was allowed to proceed for 10 minutes at room temperature prior to termination by the addition of 70 μL of CytoPhos reagent. Following an incubation of the last 10 minutes at room temperature, reactions were quantified by reading the absorbance at 650 nm in a spectrophotometer (Beckman Instruments, Inc., Model DU 530). The direct absorbance values were corrected by subtracting the absorbance of the control. The absorbance was converted to Pi concentration by comparison with a standard Pi curve. The percentage of inhibition was calculated from the concentration Pi according to the following formula:% Inhibition = (untreated - treated) / untreated x 100. The arithmetic mean was generated from the percentage of inhibition of experimental replicates. The results are shown in Table 4. Table 4 Percentage of inhibition of kinesin engine activity (n = 4) Chlorpromazine [μM] 3 2 4 8 16 32 64 Average -5. 51 -11.18 17. 42 52.91 85 82 97.79 104 .54 Desv. Standard 11. 87 25.94 17. 54 6.99 10 84 6.40 10. 96 Other phenothiazines capable of reducing the biological activity of mitotic kinesin include promethazine, thioridazine, trifluoperazine, perphenazine, fluphenazine, clozapine, prochlorperazine. The combination of chlorpromazine and pentamidine reduces the proliferation of cells ± v ± tro The ability of pentamidine (a protein tyrosine phosphatase inhibitor) and chlorpromazine (a mitotic kinesin inhibitor), in combination, to reduce cell proliferation in vitro was determined . The HCT116 human colon adenocarcinoma cell line (ATCC # CCL-247) was grown at 37 ± 5 ° C and 5% C02 in DMEM supplemented with 10% FBS, 2 mM glutamine, 1% penicillin and 1% streptomycin. The anti-proliferation assays were carried out in 384-well plates. 10X base solutions (6.6 μL) from the combination matrices were added to 40 μL of culture medium in test wells. The tumor cells were released from the culture flask using a 0.25% trypsin solution. The cells were diluted in culture medium such that 3,000 cells were delivered in 20 μL of medium to each test well. The test plates were incubated for 72-80 hours at 37 + 5 ° C with 5% C02. Twenty-five microliters of 20% Blue Alamar heated to 37 ± 5 ° C were added to each test well following the incubation period. The metabolism of Alamar Blue was quantified by the amount of fluorescence intensity 3.5-5.0 hours after the addition. The quantification, using an LJL Analyst AD reader (LJL Biosystems), was taken in the middle of the well with high attenuation, a reading time of 100 msec, an excitation filter at 530 nm, and an emission filter at 575 nm. For some experiments, the quantification was carried out using a Wallac Victor2 reader. Measurements were taken at the top of the well with stabilized energy lamp control; a reading time of 100 msec, an excitation filter at 530 nm, and an emission filter at 590 nm. No significant differences between plate readers were measured. The percent inhibition (% I) for each well was calculated using the following formula:% I = [(untreated wells - treated well) / (untreated wells prom.)] X 100 The value of average untreated wells (Wells not treated prom.) is the arithmetic mean of 40 wells of the same test plate treated with vehicle only. Negative inhibition values result from local variations in treated wells as compared to untreated wells. The data, expressed as percent inhibition, are shown in Table 5.

Table 5 Other Forms of Realization All publications and patents mentioned in the above specification are incorporated herein by reference. Various modifications and variations of the described method and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. In fact, various modifications of the described modes of carrying out the invention that are obvious to those skilled in the art of oncology or related fields are intended to be within the scope of the invention.

Claims (75)

1. CLAIMS 1. A composition comprising a first agent that reduces the biological activity of mitotic kinesin and a second agent that reduces the biological activity of protein tyrosine phosphatase, wherein said first and second agents are present in amounts that, when administered to a patient having a proliferative disease, they are sufficient to treat said disease.
2. 2. The composition of claim 1, wherein said first agent is a mitotic kinesin inhibitor.
3. The composition of claim 1, wherein said first agent is an anti-sense compound or RNAi compound that reduces the expression levels of said mitotic kinesin.
4. The composition of claim 1, wherein said first agent is a dominant negative mitotic kinesin or an expression vector encoding said dominant negative mitotic kinesin.
5. The composition of claim 1, wherein said first agent is an antibody that binds said mitotic kinesin and reduces the biological activity of the mitotic kinesin.
6. 6. The composition of any of claims 1-5, wherein said mitotic kinesin is HsEg5 / KSP.
7. The composition of claim 1, wherein said first agent is an inhibitor of aurora kinase.
8. 8. The composition of claim 1, wherein said biological activity of mitotic kinesin is enzymatic activity, motor activity, or binding activity.
9. The composition of claim 1, wherein said second agent is a protein tyrosine phosphatase inhibitor.
10. The composition of claim 1, wherein said second agent is an anti-sense compound or an RNAi compound that reduces the expression levels of said tyrosine phosphatase protein.
11. The composition of claim 1, wherein said second agent is a dominant negative tyrosine phosphatase protein or an expression vector encoding said dominant negative tyrosine phosphatase protein.
12. The composition of claim 1, wherein said second agent is an antibody that binds to said protein tyrosine phosphatase and reduces the biological activity of the protein tyrosine phosphatase.
13. The composition of any of claims 9-12, wherein said protein tyrosine phosphatase is PTP1B, PRL-1, PRL-2, PRL-3, SHP-1, SHP-2, MKP-1, MKP-2, CDC
14. 14 , CDC25A, CDC25B, or CDC25C. The composition of claim 1, wherein said second agent is a farnesyltransferase inhibitor.
15. 15. The composition of any of claims 1-14, wherein said first or second agent is present in said composition in a low dose.
16. 16. The composition of any of claims 1-14, wherein said first or second agent is present in said composition in a high dose.
17. 17. The composition of any of claims 1-16, wherein said composition is formulated for topical administration.
18. 18. The composition of any of claims 1-16, wherein said composition is formulated for systemic administration.
19. 19. A method for treating a patient having a proliferative disease, said method comprising administering to said patient a combination of: (a) a first agent that reduces the biological activity of mitotic kinesin; and (b) a second agent that reduces the biological activity of protein tyrosine phosphatase, wherein the first and second agents are administered simultaneously or within 28 days together, in amounts that together are sufficient to treat said patient.
20. The method of claim 19, wherein said first agent is a mitotic kinesin inhibitor.
21. The method of claim 19, wherein said first agent is an anti-sense compound or RNAi compound that reduces the expression levels of said mitotic kinesin.
22. 22. The method of claim 19, wherein said first agent is a dominant negative mitotic kinesin or an expression vector encoding said dominant negative mitotic kinesin.
23. 23. The method of claim 19, wherein said first agent is an antibody that binds said mitotic kinesin and reduces the biological activity of the mitotic kinesin.
24. 24. The method of any of claims 19-23, wherein said mitotic kinesin is HsEg5 / KSP.
25. 25. The method of claim 19, wherein said first agent is an aurora kinase inhibitor.
26. 26. The method of claim 19, wherein said second agent is a protein tyrosine phosphatase inhibitor.
27. The method of claim 19, wherein said second agent is an anti-sense compound or an RNAi compound that reduces the expression levels of said tyrosine phosphatase protein.
28. The method of claim 19, wherein said second agent is a dominant negative tyrosine phosphatase protein or an expression vector encoding said dominant negative tyrosine phosphatase protein.
29. 29. The method of claim 19, wherein said second agent is an antibody that binds to said protein tyrosine phosphatase and reduces the biological activity of the protein tyrosine phosphatase.
30. 30. The method of any of claims 21-25, wherein said protein tyrosine phosphatase is PTP1B, PRL-1, PRL-2, PRL-3, SHP-1, SHP-2, MKP-1, MKP-2, CDC14 , CDC25A, CDC25B, or CDC25C.
31. 31. The method of claim 19, wherein said second agent is a farnesyltransferase inhibitor.
32. 32. The method of any of claims 19-31, wherein said first and second agents are administered within 14 days of each other.
33. The method of claim 32, wherein said first and second agents are administered within 7 days of each other.
34. 34. The method of claim 33, wherein said first and second agents are administered within 1 day of each other.
35. 35. The method of any of claims 19-34, wherein said first or second agents are administered in a low dose.
36. 36. The method of any of the claims 19-34, wherein said first or second agents are administered in a high dose.
37. 37. The method of any of claims 19-36, wherein said first or second agents are administered topically or systemically.
38. 38. The method of any of claims 19-37, wherein said proliferative disease is cancer.
39. 39. The method of claim 38, wherein said cancer is selected from acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myeloma-nocitic leukemia, acute monocytic leukemia, acute erythroleukemia , chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia, polycythemia vera, Hodgkin's disease, non-Hodgkin's disease, Waldenstrom's macroglobulinemia, heavy chain disease, fibrosarcoma, myxosarcoma, liposar-coma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovium, mesothelioma, Ewing's tumor, leyomyosarcoma, rhabdomyoloma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma , carcinoma of sweat glands, gland carcinoma sebaceous cells, papillary carcinoma, papillary adenocarcinomas, cistade-nocarcinoma, medullary carcinoma, bronchiogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer , lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, meningioma, melanoma, neuroblastoma, and retinoblastoma.
40. 40. The method of claim 38, further comprising administering to said patient an anti-proliferative agent listed in Table 3.
41. 41. A method for inducing cell cycle arrest in a cell, comprising contacting the cell with a cell. first agent that reduces the biological activity of mitotic kinesin and a second agent that reduces the biological activity of protein tyrosine phosphatase.
42. 42. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) contacting proliferating cells in vitro with an agent that reduces the biological activity of mitotic kinesin and a candidate compound; and (b) determining whether the combination of the agent and the candidate compound reduces cell proliferation, in relation to the proliferation of cells contacted with the agent but not put in contact with the candidate compound, where a reduction in the proliferation of The cells identify the combination as a combination that may be useful for the treatment of a proliferative disease.
43. 43. The method of claim 42, wherein the cells are cancer cells or cells from a cancer cell line.
44. 44. A method for identifying a compound that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) providing proliferating cells designed to have reduced mitotic kinesin biological activity; (b) contacting the cells with a candidate compound; and (c) determining whether the candidate compound reduces cell proliferation, relative to cells not contacted with the candidate compound, where a reduction in cell proliferation identifies the compound as a compound that may be useful for the treatment of a proliferative disease.
45. 45. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) contacting proliferating cells in vi tro with an agent that reduces the biological activity of protein tyrosine phosphatase and a candidate compound; and (b) determining whether the combination of the agent and the candidate compound reduces cell proliferation, in relation to the proliferation of cells contacted with the agent but not put in contact with the candidate compound, where a reduction in the proliferation of The cells identify the combination as a combination that may be useful for the treatment of a proliferative disease.
46. 46. The method of claim 45, wherein the cells are cancer cells or cells from a cancer cell line.
47. 47. A method for identifying a compound that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) providing proliferating cells designed to have reduced protein tyrosine phosphatase biological activity; (b) contacting the cells with a candidate compound; and (c) determining whether the candidate compound reduces cell proliferation, relative to cells not contacted with the candidate compound, where a reduction in cell proliferation identifies the compound as a compound that may be useful for the treatment of a proliferative disease.
48. 48. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) identifying a compound that reduces the biological activity of mitotic kinesin;(b) contacting proliferating cells in vitro with an agent that reduces the biological activity of protein tyrosine phosphatase and with the compound identified in step (a); and (c) determining whether the combination of the agent and the compound identified in step (a) reduces the proliferation of cells, in relation to the proliferation of cells contacted with the agent but not contacted with the compound identified in step (a) or contacted with the compound identified in step (a) but not in contact with the agent, where a reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease.
49. 49. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) identifying a compound that reduces the biological activity of protein tyrosine phosphatase; (b) contacting proliferating cells in vitro with an agent that reduces the biological activity of mitotic kinesin and with the compound identified in step (a); and (c) determining whether the combination of the agent and the compound identified in step (a) reduces the proliferation of cells, in relation to the proliferation of cells contacted with the agent but not contacted with the compound identified in step (a) or contacted with the compound identified in step (a) but not in contact with the agent, where a reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease.
50. 50. A kit, comprising: (i) a composition comprising a first agent that reduces the biological activity of mitotic kinesin and a second agent that reduces the biological activity of protein tyrosine phosphatase; and (ii) instructions for administering said composition to a patient diagnosed with a proliferative disease.
51. 51. A kit, comprising: (i) a first agent that reduces the biological activity of mitotic kinesin; (ii) a second agent that reduces the biological activity of protein tyrosine phosphatase; and (ii) instructions for administering said first and second agents to a patient diagnosed with a proliferative disease.
52. 52. A kit comprising (i) a first agent that reduces the biological activity of mitotic kinesin and (ii) instructions for administering said agent first and a second agent that reduces the biological activity of protein tyrosine phosphatase to a patient diagnosed with a disease proliferative
53. 53. A kit comprising (i) a first agent that reduces the biological activity of tyrosine phosphatase protein and (ii) instructions for administering said agent first and a second agent that reduces the biological activity of mitotic kinesin to a patient diagnosed with a disease proliferative
54. 54. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) contacting proliferating cells in vitro with an agent that reduces the biological activity of mitotic kinesin and a candidate compound; and (b) determining whether the combination of the agent and the candidate compound reduces cell proliferation, in relation to the proliferation of cells contacted with the agent but not put in contact with the candidate compound, where a reduction in the proliferation of The cells identify the combination as a combination that may be useful for the treatment of a proliferative disease.
55. 55. The method of claim 54, wherein said agent that reduces biological activity of mitotic kinesin is a mitotic kinesin inhibitor.
56. 56. The method of claim 54, wherein said agent that reduces the biological activity of mitotic kinesin is an anti-sense compound or RNAi compound that reduces the expression levels of said mitotic kinesin.
57. 57. The method of claim 54, wherein said agent that reduces the biological activity of mitotic kinesin is a dominant negative mitotic kinesin or an expression vector encoding said dominant negative mitotic kinesin.
58. 58. The method of claim 54, wherein said agent that reduces the biological activity of mitotic kinesin is an antibody that binds said mitotic kinesin and reduces the biological activity of the mitotic kinesin.
59. 59. The method of claim 54, wherein said mitotic kinesin is HsEg5 / KSP.
60. 60. The method of claim 54, wherein said agent that reduces the biological activity of mitotic kinesin is an inhibitor of aurora kinase.
61. 61. The method of claim 54, wherein said biological activity of mitotic kinesin is enzymatic activity, motor activity, or binding activity.
62. 62. The method of claim 54, wherein the cells are cancer cells or cells of a cancer cell line.
63. 63. A method for identifying a compound that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) providing proliferating cells designed to have reduced mitotic kinesin biological activity; (b) contacting the cells with a candidate compound; and (c) determining whether the candidate compound reduces cell proliferation, relative to cells not contacted with the candidate compound, where a reduction in cell proliferation identifies the compound as a compound that may be useful for the treatment of a proliferative disease.
64. 64. The method of claim 63, wherein the cells are cancer cells or cells from a cancer cell line.
65. 65. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) contacting proliferating cells in vitro with an agent that reduces the biological activity of protein tyrosine phosphatase and a candidate compound; and (b) determining whether the combination of the agent and the candidate compound reduces cell proliferation, in relation to the proliferation of cells contacted with the agent but not put in contact with the candidate compound, where a reduction in the proliferation of The cells identify the combination as a combination that may be useful for the treatment of a proliferative disease.
66. 66. The method of claim 65, wherein said agent that reduces the biological activity of protein tyrosine phosphatase is a protein tyrosine phosphatase inhibitor.
67. 67. The method of claim 65, wherein said agent that reduces the biological activity of tyrosine phosphatase protein is an anti-sense compound or an RNAi compound that reduces the expression levels of said tyrosine phosphatase protein.
68. 68. The method of claim 65, wherein said agent that reduces the biological activity of tyrosine phosphatase protein is a dominant negative tyrosine phosphatase protein or an expression vector encoding said dominant negative tyrosine phosphatase protein.
69. 69. The method of claim 65, wherein said agent that reduces the biological activity of protein tyrosine phosphatase is an antibody that binds to said protein tyrosine phosphatase and reduces the biological activity of the protein tyrosine phosphatase.
70. 70. The method of claim 65, wherein said tyrosine phosphatase protein is PTPlB, PRL-1, PRL-2, PRL-3, SHP-1, SHP-2, MKP-1, MKP-2, CDC14, CDC25A, CDC25B. , or CDC25C.
71. 71. The method of claim 65, wherein said second agent is a farnesyltransferase inhibitor.
72. 72. The method of claim 65, wherein the cells are cancer cells or cells from a cancer cell line.
73. 73. A method for identifying a compound that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) providing proliferating cells designed to have reduced protein tyrosine phosphatase biological activity; (b) contacting the cells with a candidate compound; and (c) determining whether the candidate compound reduces cell proliferation, relative to cells not contacted with the candidate compound, where a reduction in cell proliferation identifies the compound as a compound that may be useful for the treatment of a proliferative disease.
74. 74. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) identifying a compound that reduces the biological activity of mitotic kinesin; (b) contacting proliferating cells in vitro with an agent that reduces the biological activity of protein tyrosine phosphatase and with the compound identified in step (to); and (c) determining whether the combination of the agent and the compound identified in step (a) reduces the proliferation of cells, in relation to the proliferation of cells contacted with the agent but not contacted with the compound identified in step (a) or contacted with the compound identified in step (a) but not in contact with the agent, where a reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease.
75. 75. A method for identifying a combination that may be useful for the treatment of a proliferative disease, the method comprising the steps of: (a) identifying a compound that reduces the biological activity of protein tyrosine phosphatase; (b) contacting proliferating cells in vitro with an agent that reduces the biological activity of mitotic kinesin and with the compound identified in step (a); and (c) determining whether the combination of the agent and the compound identified in step (a) reduces the proliferation of cells, in relation to the proliferation of cells contacted with the agent but not contacted with the compound identified in step (a) or contacted with the compound identified in step (a) but not in contact with the agent, where a reduction in cell proliferation identifies the combination as a combination that may be useful for the treatment of a proliferative disease.