Treatment of cancer by combination of acylofuran with ibrutinib or bortezomib

Abstract

A method for treating cancer comprises the combined use of a therapeutically effective amount of elzovtinib or an elzovtinib analog, a derivative thereof, or a pharmaceutically acceptable salt, and a therapeutically effective amount of bortezomib, ibrutinib, or an analog, derivative, or pharmaceutically acceptable salt thereof. The composition and kit are included herein. The cancer may be refractory to bortezomib and / or ibrutinib.

Description

Technical Field

[0001] The present application relates to cancer treatment, and more specifically, the present application relates to cancer treatment using a combination therapy comprising ibrutinib and / or bortezomib, or derivatives thereof. More specifically, the combination therapy can be used to treat cancers that are refractory or recurrent in subjects previously treated with ibrutinib and / or bortezomib.

Background Art

[0002] Cancer is one of the most common causes of death in humans. The development of treatment strategies for patients with advanced cancer has significantly improved overall survival. However, resistance to anticancer drugs is inevitable, and the prognosis of advanced cancer remains poor. There are several potential causes of cancer drug resistance, including changes in drug transporters, suppression of apoptosis, mitochondrial changes, promotion of DNA damage repair, autophagy, epithelial-mesenchymal transition, and cancer stem cells (CSCs). An appropriate strategy considering the mechanism is necessary to cure cancer.

[0003] The combination therapy treatment for cancer has become more common because the advantages of attacking the disease by multiple means have been recognized. Many effective combination therapy treatments have been revealed over the past few decades, and considering the still high annual death toll due to cancer, there is still a continuing need to identify effective treatment regimens for use in anticancer treatment.

[0004] Therefore, there is always a need for improved methods for treating cancer.

Summary of the Invention

[0005] This application discloses the discovery that the treatment of cancer in a subject by the combined use of irinotecan or an irinotecan analog (e.g., asiflurane) and ibrutinib and / or bortezomib (or derivatives thereof) has an effect greater than that provided by either asiflurane or ibrutinib and / or bortezomib treatment alone.

[0006] One aspect of this application includes a combination therapy for treating cancer. In an embodiment, the therapy includes administering a combination of active agents including irinotecan or an irinotecan analog (e.g., asiflurane) and ibrutinib and / or bortezomib. The method includes administering to a subject in need of treatment a combination of active agents having a therapeutically effective amount of irinotecan or an irinotecan analog thereof, a derivative or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of ibrutinib and / or bortezomib or an analog, derivative, or pharmaceutically acceptable salt thereof. The subject may have recurrent cancer and / or refractory cancer. The subject may have been previously treated with ibrutinib and / or bortezomib. The subject may have been treated in combination after the cancer has become refractory and resistant to ibrutinib and / or bortezomib.

[0007] One aspect includes a method in which the subject relapsed about 1 month, 2 months, 3 months, 4 months, or more after discontinuation of treatment with ibrutinib and / or bortezomib.

[0008] Another aspect includes a method in which the irinotecan analog is asiflurane.

[0009] Another aspect includes a method in which the irinotecan analog is hydroxyurea methyl asiflurane.

[0010] Another aspect includes a method in which the irinotecan analog has the following structure.

[0011]

Chemical formula

[0012] Another aspect includes a method in which the erdin analogue has the following structure.

[0013]

Chemical formula

[0014] Another aspect includes a method in which the erdin analogue is irofulven.

[0015] Another aspect of the present application provides a pharmaceutical composition for treating or preventing cancer, which mainly contains erdin or an erdin analogue (e.g., acylfulven) and ibrutinib and / or bortezomib or pharmaceutically acceptable salts thereof for a person who has been previously treated with ibrutinib and / or bortezomib, and is mixed with a pharmaceutically suitable carrier or excipient. The pharmaceutical composition can also be administered simultaneously, continuously, or alternately in combination with other therapeutic agents or treatment modalities.

[0016] Another aspect of the present application includes a therapy containing acylfulven, which is (+)-hydroxyurea methyl acylfulven.

[0017] Another aspect of the present application includes the treatment of cancer, which may include solid tumors and hematological malignancies. Tumors, such as, but not limited to, hyperplastic diseases or neoplastic diseases, such as breast cancer, colon cancer, rectal cancer, endometrial cancer, gastric cancer, prostate cancer, or brain cancer, mesothelioma, ovarian cancer, lung cancer, or pancreatic cancer, can be targeted for therapy, including cancer tumors, sarcomas, or mixed cancers.

Brief description of the drawings

[0018]

Figure 1

Figure 2A

Figure 2B

Figure 2C

DETAILED DESCRIPTION OF THE INVENTION

[0019] The present application provides a combination therapy for treating solid cancers and blood cancers. In embodiments, the therapy mainly comprises administering a combination comprising elzovant or an elzovant analog (e.g., acylfulvene) and ibrutinib and / or bortezomib to a person having, for example, recurrent cancer or refractory cancer and who has been treated with ibrutinib and / or bortezomib. In other embodiments, the therapy comprises administering a combination of other therapies. In other embodiments, the combination therapy can be used to treat the biochemical occurrence or recurrence of solid cancers (e.g., lung cancer, breast cancer, ovarian cancer, prostate cancer, colon cancer, rectal cancer, and bladder cancer), glioblastoma and atypical teratoid / rhabdoid tumors, and renal cell cancer). In other embodiments, the therapy comprises a combination therapy that can be used to treat the biochemical occurrence and recurrence of blood cancers, wherein acylfulvene (e.g., hydroxyurea methyl acylfulvene) or a salt thereof and ibrutinib and / or bortezomib are administered to a patient in a therapeutically effective amount. In certain embodiments, the combination can provide treatment for lymphomas such as mantle cell lymphoma (MCL) and double-hit lymphoma (DHL). In multiple myeloma (MM), the overgrowth of plasma cells in the bone marrow can displace normal hematopoietic cells.

[0020] Elzovant or acylfulvene In one embodiment, the present application includes the use of illudin or an illudin analog (e.g., acylfulvene). Acylfulvene is a class of cytotoxic semi-synthetic derivatives of illudin, a natural product that can be extracted from Omphalotus olearius. Acylfulvene, which is derived from sesquiterpene illudin S by treatment with acid (the reverse Prince reaction), is much less reactive towards thiols than illudin S.

[0021] In one example, the acylfulvene is (-)-hydroxyurea methyl acylfulvene (referred to as LP-184 of Lantern Pharma Inc.), which negatively shifts the light as shown below.

[0022]

Chemical formula

[0023] In another example, the acylfulvene is (+)-hydroxyurea methyl acylfulvene (referred to as LP-284 of Lantern Pharma Inc.), which positively shifts the light as shown below.

[0024]

Chemical formula

[0025] (+)-Hydroxyurea methyl acylfulvene and (-)-hydroxyurea methyl acylfulvene are enantiomers and are currently known.

[0026] In another example, the acylfulvene is irofulvene.

[0027] Bortezomib Bortezomib belongs to a class of agents called anti - tumor agents. It acts by killing cancer cells. Bortezomib is sold under the trade name Velcade among others and is an anticancer drug used to treat multiple myeloma and mantle cell lymphoma.

[0028] Bortezomib ([(1R)-3 - methyl - 1 - ({(2S)-3 - phenyl - 2 - [(pyrazin - 2 - ylcarbonyl)amino]propanoyl}amino)butyl]boronic acid) is a proteasome inhibitor having the structure shown below. Bortezomib is available from Janssen. The term "bortezomib" includes its pharmaceutically acceptable salts, solvates, and hydrates.

[0029]

Chemical formula

[0030] A stable liquid pharmaceutical composition of bortezomib is described in International Publication No. WO 2016 / 166653, and any such composition can be used in the methods, compositions, and uses of the present invention.

[0031] In some embodiments, the composition containing bortezomib is a "ready - to - use" formulation containing bortezomib in a dissolved or solubilized form and is intended to be used as is or further diluted with an intravenous diluent. In preferred embodiments, the pharmaceutical composition containing bortezomib is formulated for parenteral administration (e.g., injection or infusion).

[0032] Bortezomib can be administered in any suitable pharmaceutical form. For example, bortezomib can be provided as a pharmaceutical composition containing bortezomib, or a salt, solvate, or hydrate thereof, together with a pharmacologically (or pharmaceutically) acceptable excipient.

[0033] Suitable solvents include, but are not limited to, aqueous and non-aqueous solvents such as glycerin, ethanol, n-propanol, n-butanol, isopropanol, ethyl acetate, dimethyl carbonate, acetonitrile, dichloromethane, methyl ethyl ketone, methyl isobutyl ketone, cyclohexane, dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), 1,3-dimethyl-2-imidazolidinone (DMI), acetone, tetrahydrofuran (THF), dimethylformamide (DMF), propylene carbonate (PC), dimethyl isosorbide, water, and mixtures thereof. Preferred solvents are ethanol, glycerin, and water.

[0034] The bortezomib formulation for use in the present invention may contain stabilizers such as sugars and amino acids. Suitable stabilizers include glucose, trehalose, sucrose, mannitol, sorbitol, arginine, glycine, proline, methionine, lysine, and the like.

[0035] The bortezomib formulation for use in the present invention may contain a chelating agent. Suitable chelating agents include DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DTPA (diethylene triaminepentaacetic acid), EDTA (Ethylenediaminetetraacetic acid), ODDA (1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7), TTTA (1,7,13-triaza-4,10,16-trioxacyclooctadecane-N,N',N”-triacetate), DOTRP (tetraethylene glycol-1,5,9-triazacyclododecane-N,N’,N’’-tris(methylenephosphonic acid)), EGTA (ethylene glycol-bis(P-aminoethyl ether)-tetraacetic acid), and the like.

[0036] The bortezomib formulation for use in the present invention may also contain one or more antioxidants. Suitable antioxidants include, but are not limited to, monothioglycerol, ascorbic acid, sodium bisulfite, sodium dithionite, L-cysteine, thioglycolic acid, citric acid, tartaric acid, phosphoric acid, gluconic acid, thiodipropionic acid, and the like. The most preferred antioxidant is monothioglycerol.

[0037] The most preferred mode of administration of a combination of bortezomib and an additional drug for the treatment of cancer is bortezomib in the form of subcutaneous injection or intravenous injection.

[0038] The bortezomib injection used in accordance with the present invention is preferably in the form of a water-soluble boronic acid ester. The most preferred ester is mannitol boronic acid ester.

[0039] Boronic acid ester formulations, preferably mannitol esters, are typically in the form of sterile dry powder formulations. The powder is typically a lyophilized powder. The powder is dissolved in sterile water, typically a sterile isotonic aqueous sodium chloride solution, prior to administration.

[0040] The bortezomib formulations for use in the present invention may optionally contain other pharmaceutically acceptable adjuvants (e.g., buffers, pH adjusters, preservatives, isotonicity regulators, etc.). The lists of solvents, stabilizers, chelating agents, and antioxidants recited above may also be used in pharmaceutical compositions containing other cytotoxic agents described herein, unless otherwise specified.

[0041] The above bortezomib-based formulations may preferably contain mannitol and may be provided in injection vials under a nitrogen atmosphere or in pre-filled syringes.

[0042] Bruton's tyrosine kinase (BTK) inhibitor compounds comprising ibrutinib and pharmaceutically acceptable salts thereof In some embodiments, the Btk inhibitor compounds described herein are selective for Btk and kinases having a cysteine residue at the amino acid sequence position homologous to the amino acid sequence position of cysteine 481 in Btk. The Btk inhibitor compounds can form a covalent bond with Cys481 of Btk (e.g., via a Michael reaction).

[0043] In some embodiments, the Btk inhibitor is (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one (i.e., PCI-32765 / ibrutinib).

[0044] Another embodiment includes a pharmaceutical composition having a therapeutically effective amount of irinotecan or an irinotecan analog, derivatives thereof, or pharmaceutically acceptable salts, and a therapeutically effective amount of ibrutinib and / or bortezomib, or analogs, derivatives, or pharmaceutically acceptable salts thereof. The irinotecan analog can be hydroxyurea methyl acylfulvene.

[0045] In another embodiment, a kit for treating cancer in a subject includes a therapeutically effective amount of irinotecan or an irinotecan analog, derivatives thereof, or pharmaceutically acceptable salts, and a therapeutically effective amount of ibrutinib and / or bortezomib, or analogs, derivatives, or pharmaceutically acceptable salts thereof.

[0046] In another embodiment, the second therapeutic agent is one or more chemotherapeutic agents selected from camptothecin derivatives, paclitaxel, docetaxel, epothilone B, 5-FU, gemcitabine, oxaliplatin, cisplatinum, carboplatin, melphalan, dacarbazine, temozolomide, doxorubicin, imatinib, erlotinib, bevacizumab, cetuximab, and Raf kinase inhibitors.

[0047] In another embodiment, the second therapeutic agent is one or more chemotherapeutic agents selected from paclitaxel or cisplatinum.

[0048] The term "combination therapy" can include or can include the administration of the above-described therapeutic agents further combined with other biologically active components and non-drug therapies (e.g., surgery or radiation therapy). If the combination therapy further includes non-drug therapy, the non-drug therapy can be performed at any suitable time as long as a beneficial effect from the concurrent action of the therapeutic agent and the non-drug therapy is achieved. For example, in appropriate cases, a beneficial effect can be achieved even if the non-drug therapy is temporarily (possibly for several days or even several weeks or months) removed from the administration of the therapeutic agent.

[0049] In another aspect, the compositions or combination therapies herein, or pharmaceutically acceptable salts or solvates thereof, may be administered in combination with radiation therapy. Radiation therapy may also be administered in combination with the compositions of the invention and other chemotherapeutic agents described herein as part of a multi-drug therapy.

[0050] Combination therapy can be achieved by administering two or more agents, such as acylfulvene, ibrutinib and / or bortezomib, and one or more other therapeutic agents, each of which can be formulated and administered separately or by administering two or more agents in a single formulation. Other combinations are also encompassed by combination therapy. For example, two agents can be formulated together and administered in combination with a separate formulation containing a third agent. The two or more agents in combination therapy can be administered simultaneously, but they do not have to be. For example, administration of the first agent (or combination of agents) can precede administration of the second agent (or combination of agents) by minutes, hours, days, or weeks. Thus, the two or more agents can be administered within minutes of each other, or within 1, 2, 3, 6, 9, 12, 15, 18 or 24 hours of each other, or within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14 days of each other, or within 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks of each other. In some cases, even longer intervals are possible. In many cases, it is desirable for two or more agents used in combination therapy to be present in the patient's body simultaneously, but this is not necessary.

[0051] A method of combination therapy should or may provide a synergistic effect where the effect of the combination of a compound or other therapeutic agent is greater than the sum of the effects resulting from the administration of either the compound or the other therapeutic agent as a single agent. The synergistic effect may also be an effect that cannot be achieved by administering either the compound or the other therapeutic agent as a single agent. Examples of synergistic effects can include, but are not limited to, treating cancer by reducing tumor size, inhibiting tumor growth, or increasing the survival of a subject. The synergistic effect may also include reducing cancer cell survival, inducing cancer cell death, and inhibiting or delaying cancer cell growth.

[0052] The therapeutically effective dose can vary depending on the disease being treated, the severity of the disease, the route of administration, the age and general health of the patient, the use of excipients, the possibility of combination with other therapeutic treatments such as the use of other drugs, and the judgment of the treating physician, as recognized by those skilled in the art. For example, guidance for selecting an effective dose can be determined by referring to the prescribing information of acylfulvene or hydroxyurea methyl acylfulvene or the consideration of its journal.

[0053] As used herein, the term "effective amount" refers to the amount of an agent necessary to reduce at least one or more symptoms of a disease or disorder, and relates to the amount of a pharmacological composition sufficient to achieve a desired effect. Thus, the term "therapeutically effective amount" refers to the amount of an agent that, when administered to a typical subject, is sufficient to achieve a particular effect. An effective amount can be an amount sufficient to reduce the symptoms of a disease responsive to inhibition of ibrutinib and / or bortezomib. In the case of cancer therapy, in vivo efficacy can be measured, for example, by evaluating survival, time to disease progression (TTP), response rate (RR), duration of response, and / or quality of life. An effective amount can vary depending on the route of administration, the use of excipients, and combination with other agents, as will be appreciated by those skilled in the art. An effective amount in various situations also includes, as used herein, an amount sufficient to delay the onset of symptoms of a disease, alter the course of disease symptoms (e.g., but not limited to, slow the progression of disease symptoms), or reverse the symptoms of a disease. Thus, it is generally not practical to specify an exact "effective amount." However, in any given case, an appropriate "effective amount" can be determined by one of ordinary skill in the art using only routine experimentation.

[0054] The dosage range of administration of an agent by the methods described herein depends, for example, on the form of the agent, its potency, and the degree to which it is desirable to reduce the symptoms, markers or indicators of the conditions described herein, e.g., the desired rate of reduction of tumor growth. The dosage should not be so large as to cause harmful side effects. Generally, the dosage can vary depending on the age, condition, and sex of the patient and can be determined by one of ordinary skill in the art. The dosage can also be adjusted by the individual physician for any complication event.

[0055] As used herein, the term "therapeutically effective amount" refers to an amount of a pharmaceutical agent for treating, ameliorating or preventing a specified disease or condition, or for exhibiting a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The exact effective amount for a subject will depend upon the subject's weight, size, and health, the nature and extent of the condition, and the therapeutic agent or combination of therapeutic agents selected for administration. A therapeutically effective amount for a given situation can be determined by routine experimentation within the skill and judgment of the clinician. In a preferred embodiment, the disease or condition being treated is cancer. In another embodiment, the disease or condition being treated is a proliferative disorder.

[0056] For example, in the treatment of the conditions described herein, or for inducing the responses described herein (e.g., solid or hematological cancers), the effectiveness of the agents described herein can be determined by a skilled clinician. However, when one or more of the signs or symptoms of the conditions described herein change beneficially, other clinically acceptable symptoms improve or even resolve, or a desired response is induced by at least 10% for example, after treatment by the methods described herein, the treatment is considered, when this term is used herein, an "effective treatment". For example, effectiveness can be evaluated by measuring markers, indicators, symptoms and / or incidence rates of the condition being treated by the methods described herein, or any other measurable and appropriate parameter, such as tumor size and / or growth rate. Effectiveness can also be measured by the individual not deteriorating (i.e., the progression of the disease being halted), as evaluated by the need for hospitalization or medical intervention. Methods for measuring these indicators are known to those of skill in the art and / or are described herein. Treatment includes any treatment of a disease in an individual or animal (some non-limiting examples include humans or animals), including (1) inhibiting the disease, e.g., preventing the worsening of symptoms (e.g., pain or inflammation), or (2) reducing the severity of the disease, e.g., causing the regression of symptoms. An effective amount for the treatment of a disease means an amount sufficient, when administered to a subject in need thereof, to provide an effective treatment as defined herein for that disease. The effectiveness of an agent can be determined by evaluating physical indicators of the condition or a desired response. Monitoring the effectiveness of administration and / or treatment by measuring any one of such parameters, or any combination of parameters, is well within the ability of those of skill in the art. Effectiveness can be evaluated in an animal model of the condition described herein, for example, in the treatment of hematological cancer in a mouse model. When using an experimental animal model, the effectiveness of treatment is demonstrated when a statistically significant change in a marker, such as tumor size and / or tumor growth rate, is observed.In some embodiments, the therapeutically effective amount of hydroxyurea methyl-acylfulvene, acylfulvene, ilofulvene, or a pharmaceutically acceptable salt thereof is selected from the group consisting of 0.5 mg / day, 1 mg / day, 2.5 mg / day, 5 mg / day, 10 mg / day, 20 mg / day, 30 mg / day, 60 mg / day, 90 mg / day, 120 mg / day, 150 mg / day, 180 mg / day, 210 mg / day, 240 mg / day, 270 mg / day, 300 mg / day, 360 mg / day, 400 mg / day, 440 mg / day, 480 mg / day, 520 mg / day, 580 mg / day, 600 mg / day, 620 mg / day, 640 mg / day, 680 mg / day, and 720 mg / day.

[0057] The dosage should be adjusted according to the needs of the individual patient. For example, the dosage of ibrutinib and / or bortezomib to be administered is 1 - 120 mg (with respect to the parent compound), preferably 1 - 80 mg (with respect to the parent compound), and the dosing frequency is twice a day (BID), and the dosage of ibrutinib and / or bortezomib to be administered is 1 - 120 - 240 mg (with respect to the parent compound), preferably 60 - 120 mg (with respect to the parent compound), and the dosing frequency is once a day (QD). In some cases, it may be more preferable to apply the lower limit of the above dosage range, but in other cases, higher dosages can be used without causing harmful side effects.

[0058] Typical dosages of ibrutinib vary depending on the medical condition being treated. In most cases, the starting dosage for adult patients is 420 mg once a day, but for pediatric patients, it can range from 160 - 440 mg depending on height and weight. However, the dosage can be adjusted based on the individual patient's response and tolerance.

[0059] The term "treating" is used to include both therapeutic treatment and prophylactic treatment (reducing the likelihood of onset). Both terms mean reducing, suppressing, attenuating, shrinking, stopping, or stabilizing the onset or progression of a disease (e.g., a disease or disorder described herein), reducing the severity of the disease, or improving the symptoms associated with the disease.

[0060] Typical dosages of bortezomib depend on the medical condition being treated and the patient's body weight. For multiple myeloma and mantle cell lymphoma, the recommended dosage is 1.3 mg / m2 twice a week for 2 weeks, followed by a 10-day rest period. This 21-day cycle is repeated up to 8 cycles. However, the dosage can be adjusted based on the individual response and tolerance to the treatment.

[0061] The pharmaceutical composition can be contained in a container, pack, or dispenser, together with instructions for administration.

[0062] The compositions of the present invention can further form salts. The compositions of the present invention can form more than one salt per molecule, for example, mono-salts, di-salts, tri-salts. All of these forms are also considered to be within the scope of the claimed invention.

[0063] As used herein, "pharmaceutically acceptable salts" refers to derivatives of the compounds of the present invention in which the parent compound has been modified by making its acidic or basic salts. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues (e.g., amines), and alkali or organic salts of acidic residues (e.g., carboxylic acids). Pharmaceutically acceptable salts include, for example, conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include, but are not limited to, 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonate, citric acid, edetic acid, ethanedisulfonic acid, 1,2-ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, glycollyarsanilic acid, hexylresorcinol, hydrabamic acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, hydroxymaleic acid, hydroxynaphthoic acid, isethionic acid, lactic acid, lactobionic acid, laurylsulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, napsylic acid, nitric acid, oxalic acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicyclic acid, stearic acid, subacetic acid, succinic acid, sulfamic acid, sulfanilic acid, sulfuric acid, tannic acid, tartaric acid, toluenesulfonic acid, and those derived from inorganic and organic acids selected from commonly occurring amino acids such as glycine, alanine, phenylalanine, arginine, etc.

[0064] Other examples of pharmaceutically acceptable salts include hexanoic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, muconic acid, and the like. The present invention also includes salts formed when an acidic proton in the parent compound is replaced by a metal ion (e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion), or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, or the like.

[0065] All references to pharmaceutically acceptable salts are to be understood to include solvate forms (solvates) of the same salts.

[0066] As used herein, the term "selectively" means having a tendency to occur at a higher frequency in one population than in another. The populations being compared can be cell populations. Preferably, if an event occurs at a frequency more than 2-fold higher in population A compared to population B, it is said to occur selectively in population A relative to population B. An event occurs selectively if it occurs at a frequency more than 5-fold higher in population A. An event occurs more preferably selectively if it occurs at a frequency more than 10-fold higher in population A compared to population B, even more preferably if it occurs at a frequency more than 50-fold higher, still more preferably if it occurs at a frequency more than 100-fold higher, and most preferably if it occurs at a frequency more than 1000-fold higher. For example, cell death is said to occur selectively in cancer cells if it occurs more frequently than 2-fold in cancer cells compared to normal cells.

[0067] The composition, or a pharmaceutically acceptable salt or solvate thereof, is administered orally, nasally, transdermally, by the pulmonary, inhalation, buccal, sublingual, intraperitoneal, subcutaneous, intramuscular, intravenous, rectal, intrapleural, intrathecal, and parenterally. In one embodiment, the compound is administered orally. Those skilled in the art will recognize the advantages of the particular route of administration.

[0068] The dosage regimen utilizing the compounds is selected according to a variety of factors including the type, species, age, weight, sex and medical condition of the patient, the severity of the condition being treated, the route of administration, the renal and hepatic function of the patient, and the particular compound or salt thereof being employed. A ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, reverse, or arrest the progress of the condition.

[0069] Techniques for formulation and administration of the disclosed compounds of the invention can be found in Remington: the Science and Practice of Pharmacy, 19.sup.th edition, Mack Publishing Co., Easton, Pa. (1995). In one embodiment, the compounds described herein and their pharmaceutically acceptable salts are used in pharmaceutical preparations in combination with a pharmaceutically acceptable carrier or diluent. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile aqueous or organic solutions. The compounds are present in such pharmaceutical compositions in an amount sufficient to provide the desired dosage within the ranges described herein.

[0070] All percentages and ratios used herein are by weight unless otherwise indicated. Other features and advantages of the invention will be apparent from the different examples. The examples provided illustrate different components and methodologies useful in practicing the invention. The examples do not limit the claimed invention. Based on this disclosure, those skilled in the art can identify and use other components and methodologies useful in practicing the invention.

[0071] As used herein, a "subject in need thereof" is a subject having a pre-cancerous condition. Preferably, the subject in need thereof has cancer. The "subject" includes mammals. The mammal can be, for example, any mammal, such as a human, primate, bird, mouse, rat, dog, cat, cow, horse, goat, camel, sheep or pig. Preferably, the mammal is a human. The subjects of the present invention include any human subject diagnosed with cancer or a pre-cancerous condition, having symptoms of cancer or a pre-cancerous condition, or at risk of developing cancer or a pre-cancerous condition.

[0072] The subject in need thereof can have refractory cancer or resistant cancer. "Refractory cancer or resistant cancer" means cancer that does not respond to treatment. The cancer can be resistant at the start of treatment or become resistant during treatment. In some embodiments, the subject in need thereof has cancer recurrence after remission with a recent therapy. In some embodiments, the subject in need thereof has received all known effective therapies for cancer treatment and failed. In some embodiments, the subject in need thereof has previously received at least one therapy. In certain embodiments, the previous therapy is a monotherapy. In certain embodiments, the previous therapy is a combination therapy.

[0073] "Recurrent cancer" is cancer that has been treated before and as a result of that treatment, the subject has fully or partially recovered (i.e., the subject is said to be in a remission state), but the cancer has recurred or worsened after the treatment has been discontinued. In other words, recurrent cancer is cancer that has become resistant to treatment after a period during which the treatment was effective and the subject has fully or partially recovered. Typically, in patients with multiple myeloma, the onset of resistant and refractory disease occurs after multiple rounds of treatment. Additionally, ibrutinib can become refractory and lead to treatment resistance when cancer cells develop mutations that bypass the drug's mechanism of action or alter alternative signaling pathways. This can occur at various stages of treatment and can vary depending on the type of cancer being treated and the individual response of the patient to the drug.

[0074] In some embodiments, the subject in need thereof may have secondary cancer as a result of previous therapy. "Secondary cancer" means cancer that results from or is caused by a previous carcinogenic therapy such as chemotherapy.

[0075] Cancer is a group of diseases that can cause almost any sign or symptom. The signs and symptoms depend on where the cancer is, the size of the cancer, and how much it affects nearby organs or structures. If the cancer spreads (metastasizes), symptoms can appear in different parts of the body.

[0076] Treatment of cancer can reduce the size of the tumor. A reduction in the size of the tumor may also be referred to as "tumor regression". Preferably, after treatment, the tumor size is reduced by 5% or more relative to its size before treatment, more preferably the tumor size is reduced by 10% or more, more preferably by 20% or more, more preferably by 30% or more, more preferably by 40% or more, even more preferably by 50% or more, most preferably by 75% or more. The size of the tumor may be measured by any reproducible measuring means. The size of the tumor may be measured as the diameter of the tumor.

[0077] Treatment of cancer reduces the number and size of tumors. Preferably, after treatment, the number or size of the tumors is reduced by 5% or more relative to the number before treatment, more preferably the number or size of the tumors is reduced by 10% or more, more preferably by 20% or more, more preferably by 30% or more, more preferably by 40% or more, even more preferably by 50% or more, most preferably by more than 75%. The number of tumors may be measured by any reproducible measuring means. The number of tumors may be measured by counting the tumors that are visible to the naked eye or at a specific magnification. Preferably, the specific magnification is 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, or 50-fold.

[0078] Treating cancer can result in a decrease in the number of metastatic lesions in other tissues or organs distant from the primary tumor site. Preferably, after treatment, the number of metastatic lesions is reduced by 5% or more, more preferably 10% or more, more preferably 20% or more, more preferably 30% or more, more preferably 40% or more, even more preferably 50% or more, and most preferably more than a 75% decrease compared to the number before treatment. The number of metastatic lesions may be measured by any reproducible measurement means. The number of metastatic lesions can be measured by counting the metastatic lesions that are visible to the naked eye or at a specific magnification. Preferably, the specific magnification is 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, or 50-fold.

[0079] Treating cancer can increase the average survival time of the treated population of subjects compared to a population receiving the carrier alone. Preferably, the average survival time increases by more than 30 days, more preferably more than 60 days, more preferably more than 90 days, and most preferably more than 120 days. The increase in the average survival time of the population may be measured by any reproducible means. The increase in the average survival time of the population can be measured, for example, by calculating the average length of survival for the population after the start of treatment with the active compound. The increase in the average survival time of the population can also be measured, for example, by calculating the average length of survival for the population after the completion of the first round of treatment with the active compound.

[0080] Treatment of cancer can result in an increase in the average survival period of the treated population compared to the untreated population. Preferably, the average survival period increases by more than 30 days, more preferably by more than 60 days, more preferably by more than 90 days, and most preferably by more than 120 days. The increase in the average survival period of the population may be measured by any reproducible means. The increase in the average survival period of the population can be measured, for example, by calculating the average length of survival for the population after the start of treatment with the active compound. The increase in the average survival period of the population can also be measured, for example, by calculating the average length of survival for the population after the completion of the first round of treatment with the active compound.

[0081] Treatment of cancer can result in an increase in the average survival period of the treated population compared to a population receiving monotherapy with a compound of the invention or a drug that is not a pharmaceutically acceptable salt or solvate thereof. Preferably, the average survival period increases by more than 30 days, more preferably by more than 60 days, more preferably by more than 90 days, and most preferably by more than 120 days. The increase in the average survival period of the population may be measured by any reproducible means. The increase in the average survival period of the population can be measured, for example, by calculating the average length of survival for the population after the start of treatment with the active compound. The increase in the average survival period of the population can also be measured, for example, by calculating the average length of survival for the population after the completion of the first round of treatment with the active compound.

[0082] Treating cancer can result in a reduction in the mortality rate of the population being treated as compared to a population receiving the carrier alone. Treating cancer can result in a reduction in the mortality rate of the population being treated as compared to an untreated population. Treating cancer can result in a reduction in the mortality rate of the population being treated as compared to a population receiving monotherapy with a compound of the invention or a drug that is not a pharmaceutically acceptable salt or solvate thereof. Preferably, the mortality rate is reduced by more than 2%, more preferably by more than 5%, more preferably by more than 10%, and most preferably by more than 25%. The reduction in the mortality rate of the population being treated may be measured by any reproducible means. The reduction in the mortality rate of the population can be measured, for example, by calculating the mean number of disease-related deaths per unit time for the population after the start of treatment with the active compound. The reduction in the mortality rate of the population can also be measured, for example, by calculating the average number of disease-related deaths per unit time for the population after completion of the first round of treatment with the active compound.

[0083] Treating cancer can result in a reduction in the tumor growth rate. Preferably, after treatment, the tumor growth rate is reduced by at least 5% relative to the pre-treatment figure, more preferably the tumor growth rate is reduced by at least 10%, more preferably by at least 20%, more preferably by at least 30%, more preferably by at least 40%, more preferably by at least 50%, even more preferably by at least 50%, and most preferably by at least 75%. The tumor growth rate may be measured by any reproducible measurement means. The tumor growth rate can be measured by the change in the diameter of the tumor per unit time.

[0084] Treatment of cancer can result in a reduction in tumor regrowth. Preferably, after treatment, tumor regrowth is less than 5%, more preferably less than 10%, more preferably less than 20%, more preferably less than 30%, more preferably less than 40%, more preferably less than 50%, even more preferably less than 50%, and most preferably less than 75%. Tumor regrowth may be measured by any reproducible measurement means. Tumor regrowth is measured, for example, by measuring the increase in the diameter of the tumor after previous tumor shrinkage following treatment. Reduction in tumor regrowth is indicated by the non-recurrence of the tumor after treatment is stopped.

[0085] Treatment or prevention of a cell proliferative disorder can result in a decrease in the rate of cell proliferation. Preferably, after treatment, the rate of cell proliferation is decreased by at least 5%, more preferably at least 10%, more preferably at least 20%, more preferably at least 30%, more preferably at least 40%, more preferably at least 50%, even more preferably at least 50%, and most preferably at least 75%. The rate of cell proliferation may be measured by any reproducible measurement means. The rate of cell proliferation is measured, for example, by measuring the number of dividing cells in a tissue sample per unit time.

[0086] Treatment or prevention of a cell proliferative disorder can result in a decrease in the proportion of proliferating cells. Preferably, after treatment, the proportion of proliferating cells is decreased by at least 5%, more preferably at least 10%, more preferably at least 20%, more preferably at least 30%, more preferably at least 40%, more preferably at least 50%, even more preferably at least 50%, and most preferably at least 75%. The proportion of proliferating cells may be measured by any reproducible measurement means. Preferably, the proportion of proliferating cells is measured, for example, by quantifying the number of dividing cells relative to the number of non-dividing cells in a tissue sample. The proportion of proliferating cells may be equivalent to the mitotic index.

[0087] The treatment or prevention of a cell proliferative disorder can result in a decrease in the size of the area or zone of cell proliferation. Preferably, after treatment, the size of the area or zone of cell proliferation is reduced by at least 5% relative to its size before treatment, more preferably at least 10% reduction, more preferably at least 20% reduction, more preferably at least 30% reduction, more preferably at least 40% reduction, more preferably at least 50% reduction, even more preferably at least 50% reduction, most preferably at least 75% reduction. The size of the area or zone of cell proliferation may be measured by any reproducible measuring means. The size of the area or zone of cell proliferation may be measured as the diameter or width of the area or zone of cell proliferation.

[0088] The treatment or prevention of a cell proliferative disorder can result in a decrease in the number or proportion of cells having abnormal appearance or geometry. Preferably, after treatment, the number of cells having abnormal morphology is reduced by at least 5% relative to its size before treatment, more preferably at least 10% reduction, more preferably at least 20% reduction, more preferably at least 30% reduction, more preferably at least 40% reduction, more preferably at least 50% reduction, even more preferably at least 50% reduction, most preferably at least 75% reduction. The appearance or morphology of abnormal cells may be measured by any reproducible measuring means. The geometry of abnormal cells can be measured, for example, by using an inverted tissue culture microscope. The geometry of abnormal cells can take the form of nuclear pleomorphism.

[0089] Administering the composition of the present invention to a cell or subject in need thereof can result in the modulation (i.e., stimulation or inhibition) of the activity of the protein methyltransferase of interest.

[0090] By treating cancer or a cell proliferative disorder, cell death can be induced, preferably, the cell death results in at least a 10% decrease in the number of cells in the population. More preferably, the cell death is at least a 20% decrease, more preferably, at least a 30% decrease, more preferably, at least a 40% decrease, more preferably, at least a 50% decrease, most preferably, at least a 75% decrease. The number of cells in the population may be measured by any reproducible means. The number of cells in the population can be measured by fluorescence activated cell sorting (FACS), immunofluorescence microscopy and light microscopy. The method for measuring cell death is as shown in Li et al., Proc. Natl. Acad. Sci. USA. 100(5):2674-8, 2003. In one embodiment, the cell death is caused by apoptosis.

[0091] Preferably, an effective amount of the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, is not significantly cytotoxic to normal cells. A therapeutically effective amount of a compound is not significantly cytotoxic to normal cells if administration of the compound at a therapeutically effective amount does not induce more than 10% cell death of normal cells. A therapeutically effective amount of a compound does not significantly affect the survival rate of normal cells if administration of the compound at a therapeutically effective amount does not induce more than 10% cell death of normal cells. In one embodiment, the cell death is caused by apoptosis.

[0092] By contacting cells with the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, selective induction or activation of cell death can be achieved in cancer cells. By administering the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, to a subject in need thereof, selective induction or activation of cell death can be achieved in cancer cells. By contacting cells with the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, selective induction of cell death can be achieved in one or more cells affected by a cell proliferative disorder. Preferably, by administering the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, to a subject in need thereof, selective induction of cell death is achieved in one or more cells affected by a cell proliferative disorder.

[0093] The present invention provides a method for treating or preventing cancer by administering the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, to a subject in need thereof, wherein the administration of the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, results in prevention of cancer cell proliferation by accumulation of cells in one or more phases of the cell cycle (e.g., G1, G1 / S, G2 / M), or induction of cellular senescence, or promotion of tumor cell differentiation, or promotion of cell death in cancer cells by cytotoxicity, necrosis or apoptosis without significant amounts of cell death in normal cells, or anti-tumor activity with a therapeutic index of at least 2. As used herein, "therapeutic index" is the ratio of the maximum tolerated dose to the effective dose.

[0094] The term "kit" means that the combination partners defined above can be administered either independently or by use of different fixed combinations having an identified amount of the combination partner, i.e., simultaneously or at different times. Subsequently, the parts of the kit of parts can be administered, for example, simultaneously or shifted over time, i.e., at different times for any of the parts of the kit of parts, at equal or different time intervals. The ratio of the total amounts of the combination partners administered in the combination preparation can vary. The combination partners can be administered by the same route or different routes.

[0095] One of ordinary skill in the art can refer to general reference texts for a detailed description of the known art or equivalent art discussed herein. These texts can, of course, also be referred to when making or using aspects of the present invention.

Examples

[0096] In order for the present disclosure disclosed herein to be more efficiently understood, examples are presented below. It should be understood that these examples are for illustrative purposes only and should in no way be construed as limiting the present disclosure.

[0097] Example 1 The effects of LP-284 and of ibrutinib and / or bortezomib on the growth of MCL xenograft tumors are shown in FIGS. 1 and 2A. Xenograft mice (n = 8 / treatment group) derived from the JeKo-1 MCL cell line were treated on the days indicated by the arrows in the graph with vehicle, LP-284 (2 mg / kg or 4 mg / kg, i.v.), bortezomib (1 mg / kg, i.p.), or ibrutinib (50 mg / kg, p.o.). Tumor volume was measured every 2-4 days after the start of treatment. FIG. 1 shows that the cancer cells become refractory and recur several days after treatment. Treatment with low doses of LP-284, ibrutinib, and / or bortezomib each resulted in refractory cancer after initially controlling cell growth in the tumors.

[0098] As shown in FIGS. 2A and 2C in the same study, LP-284, at a size of 2000 mm3, can almost completely eliminate these lymphomas that are refractory to bortezomib or ibrutinib (FIG. 2A). There are multiple resistance mechanisms for these two drugs, such as mutations and dysregulation of proteasome subunits, BTK mutations, upregulated NF-κB, and cell survival pathways. These results indicate that LP-284 treatment can restore the treatment plan for patients refractory to bortezomib or ibrutinib.

[0099] Example 2 FIGS. 2A, 2B, and 2C show that LP-284 with bortezomib or ibrutinib reduces bortezomib-refractory and ibrutinib-refractory MCL xenograft tumors. JeKo-1 MCL xenograft tumors were refractory to bortezomib and ibrutinib under the dosing regimen as shown in FIG. 2A, resulting in a tumor volume of approximately 2000 mm3 on day 17. The two drugs were replaced with 1 cycle of 4 mg / kg LP-284 or vehicle (saline). Bortezomib- and ibrutinib-refractory mice were stratified into two groups of 4 mice each based on their tumor size, and then they were treated with LP-284 or vehicle. Vehicle-treated tumors continued to grow until all mice died or were sacrificed due to excessive tumor size, while 1 cycle of 4 mg / kg LP-284 resulted in almost complete tumor regression (FIGS. 2A and 2B). In addition, LP-284 provided a survival period approximately 2-fold longer (p < 0.001) than the vehicle in xenograft mice pre-treated with these bortezomib or ibrutinib and refractory (FIG. 2C). No significant weight loss at the end of treatment (EOT) was observed in any of the treatment groups (one-sided t-test, p > 0.05).

[0100] LP-284 rescues bortezomib- and ibrutinib-refractory MCL xenograft mice. Xenograft mice derived from the JeKo-1 MCL cell line were treated with bortezomib (1 mg / kg, i.p.) or ibrutinib (50 mg / kg, p.o.) until day 16 when the tumor size reached approximately 2000 mm3. Then, each treatment group was further divided into two smaller treatment groups (n = 4 / smaller treatment group), and as indicated by the arrows in the graph, one smaller treatment group was treated with vehicle and the other was treated with 4 mg / kg of LP-284 (i.v.). Figure 2A shows that the tumor volume decreased rapidly in the LP-284 treatment group. Figure 2B shows representative photographs of large tumors (top) in mice after switching to vehicle treatment and almost complete tumor regression (bottom) in mice after switching to LP-284 treatment. Figure 2C shows a Kaplan–Meier survival curve indicating an extension of the survival period in the group rescued by LP-284.

[0101] Methodology In vivo animal study Five-week-old NOD.Scid mice were procured from Envigo (Item 17003F). The mice were given Teklad irradiated (sterilized) mouse diet and bedded with Teklad irradiated (sterilized) corn cob bedding from Envigo (Indianapolis, IN). The mice were housed in an Optimice carousel sterilization compartment with filtered air supply in disposable cages from Animal Care Systems, Inc. (Centennial, CO). A total of 1 × 10 7 cells of JeKo-1 cells were subcutaneously injected into the right posterior flank of each animal using a 27-gauge needle. Vehicle (saline), 4 mg / kg or 2 mg / kg of LP-284 was administered as intravenous injections on days 1, 3, 5, 7, 9, 17, 19, 21, 23, 25. 50 mg / kg of ibrutinib was administered orally daily from days 1 to 16. 1 mg / kg of bortezomib was administered as intraperitoneal injections on days 1, 4, 7, 10. The ibrutinib- and bortezomib-treated groups were further treated intravenously with vehicle or 4 mg / kg of LP-284 on days 17, 19, 21, 23, 25. Tumors were measured two-dimensionally using calipers and the volume was calculated using the following formula. Tumor volume (mm 3 ) = w2×l / 2 where w = the width of the tumor, l = the length of the tumor, in mm. For this study, calipers were aligned at the tumor margin (the tumor was not compressed by the calipers).

[0102] Although several exemplary aspects and embodiments have been discussed above, those skilled in the art will recognize certain changes, permutations, additions, and partial combinations thereof. Accordingly, the following appended claims and subsequent claims are intended to be construed to include all such changes, permutations, additions, and partial combinations as fall within their true spirit and scope.

Claims

1. A combination pharmaceutical formulation for the treatment of ibrutinib-refractory or ibrutinib-resistant cancer, comprising a therapeutically effective amount a. Hydroxyureamethylacylfluben, or a pharmaceutically acceptable salt thereof, and b. Ibrutinib, or a pharmaceutically acceptable salt thereof. A combination of pharmaceuticals including this.

2. The combination drug formulation according to claim 1, wherein the cancer has been previously treated with ibrutinib.

3. The combination of pharmaceuticals according to claim 1, wherein the cancer has recurred during or after previous treatment with ibrutinib.

4. The combination drug formulation according to claim 1, wherein the cancer is refractory to ibrutinib and resistant to ibrutinib.

5. The combination of pharmaceuticals according to claim 1, wherein the cancer becomes resistant to ibrutinib during treatment with ibrutinib.

6. The combination drug formulation according to claim 1, wherein the subject is treated after discontinuation of previous ibrutinib treatment.

7. The combination drug formulation according to claim 6, wherein the cancer recurred more than one month after discontinuation of the previous ibrutinib treatment.

8. The pharmaceutical combination formulation according to claim 1, wherein the hydroxyurea methylacylflubene is (+)-hydroxyurea methylacylflubene.

9. The pharmaceutical combination formulation according to claim 1, wherein the hydroxyurea methylacylflubene is (-)-hydroxyurea methylacylflubene.

10. The pharmaceutical combination formulation according to claim 1, wherein the (+)-hydroxyureamethylacylfluben is formulated for intravenous administration and the ibrutinib is formulated for intravenous or subcutaneous administration.

11. The combination pharmaceutical formulation according to claim 1, wherein the hydroxyurea methylacylfluben and the ibrutinib are formulated for separate, simultaneous, or sequential administration.

12. The pharmaceutical combination formulation according to claim 1, wherein the hydroxyurea methylacylfluben is formulated for parenteral administration and the ibrutinib is formulated for oral administration.

13. The pharmaceutical combination formulation according to claim 12, wherein the hydroxyurea methylacylfluben is formulated for intravenous administration.

14. The combination pharmaceutical preparation according to claim 1, wherein the cancer is a hematological malignancy selected from lymphoma, leukemia, multiple myeloma, and mantle cell lymphoma.

15. A kit for the treatment of ibrutinib-refractory or ibrutinib-resistant cancer, a. A first container containing hydroxyureamethylacylfluben or a pharmaceutically acceptable salt thereof, and b. A second container containing ibrutinib or a pharmaceutically acceptable salt thereof. Includes, A kit comprising the hydroxyurea methylacylfluben and ibrutinib, provided for separate, simultaneous, or sequential use.

16. A combination of pharmaceutical formulations for treating bortezomib-refractory or bortezomib-resistant cancer in a subject who has received at least two previous rounds of bortezomib treatment, comprising a therapeutically effective amount a. Hydroxyureamethylacylfluben, or a pharmaceutically acceptable salt thereof, and b. Bortezomib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. A combination of pharmaceuticals including this.

17. The combination of pharmaceutical formulations according to claim 16, wherein the at least two prior rounds of bortezomib treatment comprise at least two prior treatment cycles containing bortezomib.

18. The combination drug formulation according to claim 16, wherein the cancer is refractory to bortezomib and resistant to bortezomib.

19. The combination of pharmaceuticals according to claim 16, wherein the cancer has recurred during or after prior bortezomib treatment.

20. The combination drug formulation according to claim 16, wherein the cancer recurred more than one month after discontinuation of the previous bortezomib treatment.

21. The pharmaceutical combination formulation according to claim 16, wherein the hydroxyureamethylacylflubene is (+)-hydroxyureamethylacylflubene.

22. The pharmaceutical combination formulation according to claim 16, wherein the hydroxyurea methylacylflubene is (-)-hydroxyurea methylacylflubene.

23. The combination of pharmaceutical formulations according to claim 16, wherein mantle cell lymphoma has relapsed after an initial response to prior bortezomib treatment.

24. The combination pharmaceutical formulation according to claim 16, wherein the hydroxyurea methylacylfluben and the bortezomib are formulated for separate, simultaneous, or sequential administration.

25. The pharmaceutical combination formulation according to claim 16, wherein the hydroxyurea methylacylfluben is formulated for intravenous administration and the bortezomib is formulated for intravenous or subcutaneous administration.

26. The pharmaceutical combination formulation according to claim 16, wherein the bortezomib is formulated as an injectable formulation.

27. ​​The pharmaceutical combination formulation according to claim 16, wherein the bortezomib is provided in a formulation containing a boronic acid ester.

28. The combination pharmaceutical preparation according to claim 16, wherein the cancer is a hematological malignancy.

29. The combination pharmaceutical preparation according to claim 28, wherein the hematological malignancy is selected from multiple myeloma, mantle cell lymphoma, non-Hodgkin lymphoma, lymphoma, and leukemia.

30. A kit for treating bortezomib-refractory or bortezomib-resistant cancer in a subject who has received at least two previous rounds of bortezomib treatment, a. A first container containing hydroxyureamethylacylfluben or a pharmaceutically acceptable salt thereof, and b. A second container containing bortezomib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. Includes, A kit comprising the hydroxyurea methylacylfluben and the bortezomib, provided for separate, simultaneous, or sequential use.

31. The kit according to claim 30, wherein mantle cell lymphoma has relapsed after an initial response to prior bortezomib treatment.

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