Method for pre-treating recurrent cancer

Abstract

The present invention relates to a histone deacetylase (HDAC) inhibitor for use in pre-treatment of recurrent cancer prior to the treatment of recurrent cancer, wherein the HDAC inhibitor is selected from the group consisting of valproic acid, sodium valproate semihydrate, sodium valproate, magnesium valproate, or a mixture thereof, and the HDAC inhibitor is administered to a subject suffering from recurrent cancer.

Description

Technical Field

[0001] Field of the Invention The present invention relates to the field of methods for pre-treating recurrent cancer, i.e., as pre-treatment prior to other treatments to enhance the effectiveness of treatment or reduce side effects, in which one or more compositions are administered to an individual in need of cancer treatment.

Background Art

[0002] Background of the Invention Cancer can be defined as the abnormal growth of cells, which shows signs of uncontrolled proliferation and disrupted programmed cell death. From a classical perspective, a series of genetic events cause malignant transformation, resulting in a cell clone that does not respect the integrity of other cells and tissues and may eventually metastasize. Cancer involves every tissue of the body and can have many different forms in each location of the body.

[0003] Malignant lymphoma can be defined as the malignant transformation of hematopoietic lymphoid cells. Lymphomas can be divided into aggressive lymphomas and indolent lymphomas. Aggressive lymphomas are characterized by a rapid growth pattern and can have dramatic clinical features. However, aggressive lymphomas can be cured by chemotherapy, radiotherapy, and treatment with monoclonal antibodies. In contrast, indolent lymphomas (such as follicular lymphoma) have a slow growth pattern and usually milder clinical findings. However, indolent lymphomas cannot be cured by standard lymphoma treatments and can sometimes be treated by allogeneic stem cell transplantation. The median survival of follicular lymphoma is 8-10 years. Diffuse large B-cell lymphoma and Hodgkin lymphoma belong to the group of aggressive lymphomas, while follicular lymphoma and chronic lymphocytic leukemia are indolent lymphomas. Myeloma consists of malignant transformed plasma cells. They are related to indolent lymphomas but are usually considered to exist on their own. The prognosis is pessimistic, and the median survival is 5-7 years.

[0004] One of the most common subtypes of malignant lymphoma is diffuse large B-cell lymphoma (DLBCL), with an incidence of approximately 500 cases / year in Sweden. DLBCL constitutes 60 - 70% of aggressive lymphomas. The median age at diagnosis is 70 years, and DLBCL occurs slightly more frequently in men than in women.

[0005] The standard first-line treatment for DLBCL is chemotherapy consisting of a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP). In recent years, the addition of rituximab, a CD20 antibody, has become an international clinical standard (R-CHOP), resulting in improved progression-free survival, event-free survival, disease-free survival, and overall survival (Morrison, Expert Rev Anticancer Ther, 2008; 8(10): pp. 1651 - 1658). Nevertheless, since as many as 45% of patients still die from the disease, there is an obvious clinical need to increase progression-free survival in DLBCL patients.

[0006] One important area of cancer disease research is the control of DNA transcription. This is a complex process, and the mechanisms involved are only partially known. Histone deacetylases (HDACs) can control the expression of tumor suppressor genes and the activity of transcription factors involved in both cancer development and progression. HDACs act via changes in either DNA or chromatin components through histone deacetylation, thereby affecting the three-dimensional conformation of DNA without changing or interfering with its sequence (epigenetic modification). It has also been suggested that they can change the sensitivity to DNA-damaging chemotherapy by regulating chromatin structure. According to these, several in vitro studies have suggested that HDAC inhibition can have a synergistic effect with chemotherapy.

[0007] Accordingly, in recent years, a number of HDAC inhibitors have been developed. They can be divided into four classes: hydroxamic acid / carbamic acid, cyclic peptides, fatty acids, and benzamides. Examples of HDAC inhibitors approved for the treatment of cancer include vorinostat and romidepsin, which have been approved by the FDA (Food and Drug Administration) for the treatment of cutaneous T-cell lymphoma and are currently being evaluated for the treatment of other malignancies.

[0008] Clinically, the most well-known HDAC inhibitor is valproic acid, an anticonvulsant drug that has been used for the treatment of epilepsy since the 1970s. Valproic acid belongs to the fatty acid class of inhibitors.

[0009] The inventors have already shown, as in WO2012 / 128709, that an HDAC inhibitor in combination with a steroid is useful when administered to a human in need of cancer treatment as a pretreatment prior to other treatments, and the effect of the treatment is promoted. SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

[0010] Despite the promise of more effective treatment of cancer created by the emergence of pretreatment using the HDAC inhibitors taught in WO2012 / 128709, there are still challenges to be overcome in order to more fully realize the potential of pretreatment of cancer with HDAC inhibitors.

[0011] In particular, there is still a need for effective strategies for treating recurrent diseases, i.e., those in which the patient has experienced a first type of treatment but the cancer remains or reappears. Typically, the first type of treatment may not be appropriate for treating recurrent cancer, and this patient group requires new treatments. MEANS FOR SOLVING THE PROBLEMS

[0012] SUMMARY OF THE INVENTION The present invention relates to the research result that valproic acid and its derivatives can be administered as pre-treatment for recurrent cancer prior to treatment.

[0013] Accordingly, in a first aspect, the present invention provides a histone deacetylase (HDAC) inhibitor for use in pre-treatment of recurrent cancer prior to treatment of recurrent cancer, which is selected from the group consisting of valproic acid, valproic acid semi-sodium, sodium valproate, magnesium valproate, or a mixture thereof, and is administered to a subject suffering from recurrent cancer.

[0014] A corresponding second aspect of the present invention is a method for pre-treating recurrent cancer, wherein a histone deacetylase HDAC inhibitor selected from the group consisting of valproic acid, valproic acid semi-sodium, sodium valproate, magnesium valproate, or a mixture thereof is administered to a subject suffering from recurrent cancer.

[0015] A corresponding third aspect of the present invention relates to the use of a histone deacetylase HDAC inhibitor selected from the group consisting of valproic acid, valproic acid semi-sodium, sodium valproate, magnesium valproate, or a mixture thereof in the manufacture of a medicament for pre-treating recurrent cancer.

[0016] A fourth aspect of the present invention is a histone deacetylase (HDAC) inhibitor and a therapeutic agent for use in combination therapy of recurrent cancer, wherein the HDAC inhibitor is selected from the group consisting of valproic acid, valproic acid semi-sodium, sodium valproate, magnesium valproate, or a mixture thereof, the therapeutic agent is selected from the list consisting of bendamustine, gemcitabine, and carboplatin, and the HDAC inhibitor is administered to a subject suffering from recurrent cancer prior to the therapeutic agent administered to the subject suffering from recurrent cancer.

[0017] A corresponding fifth aspect of the present invention is a method for combination therapy of recurrent cancer, A step of pre-treating recurrent cancer by administering to a subject suffering from recurrent cancer a histone deacetylase HDAC inhibitor selected from the group consisting of valproic acid, sodium valproate semihydrate, sodium valproate, magnesium valproate, or a mixture thereof, and A step of treating recurrent cancer by administering to a subject suffering from recurrent cancer a therapeutic agent selected from the list consisting of bendamustine, gemcitabine, and carboplatin relates to a method comprising.

[0018] A sixth aspect of the present invention is a kit for pre-treating and treating recurrent cancer, comprising one or more doses of an HDAC inhibitor selected from the group consisting of valproic acid, sodium valproate semihydrate, sodium valproate, magnesium valproate, or a mixture thereof, one or more doses of a therapeutic agent selected from the list consisting of bendamustine, gemcitabine, and carboplatin relates to a kit comprising.

Brief Description of the Drawings

[0019]

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Mode for Carrying Out the Invention

[0020] Detailed Description of the Invention The first aspect of the present invention is a histone deacetylase (HDAC) inhibitor for use in the pretreatment of recurrent cancer prior to the treatment of recurrent cancer, which is selected from the group consisting of valproic acid, valproic acid semisodium, sodium valproate, magnesium valproate, or a mixture thereof, and is administered to a subject suffering from recurrent cancer.

[0021] A corresponding second aspect of the present invention is a method for pre-treating recurrent cancer, wherein a histone deacetylase HDAC inhibitor selected from the group consisting of valproic acid, valproic acid semisodium, sodium valproate, magnesium valproate, or a mixture thereof is administered to a subject suffering from recurrent cancer.

[0022] A corresponding third aspect of the present invention relates to the use of a histone deacetylase HDAC inhibitor selected from the group consisting of valproic acid, valproic acid semisodium, sodium valproate, magnesium valproate, or a mixture thereof in the manufacture of a medicament for pre-treating recurrent cancer.

[0023] Administration of the HDAC inhibitor is preferably systemic, for example by oral administration or parenteral (intravenous, subcutaneous, and intramuscular) administration. The HDAC inhibitor may be formulated in a pharmaceutical composition. The pharmaceutical composition may be in a form selected from the group consisting of granules, powders, tablets, coated tablets, microcapsules, microparticles, and foaming agent forms. The pharmaceutical composition is preferably formulated for systemic administration. As a result, the preferred routes of administration are oral administration and parenteral (intravenous, subcutaneous, and intramuscular) administration. The pharmaceutical composition may contain one or more other pharmaceutically acceptable pharmaceutical ingredients, such as pharmaceutically acceptable diluents, carriers, excipients, or buffers. "Pharmaceutically acceptable" means a non-toxic compound that does not reduce the biological activity effectiveness of the active ingredient.

[0024] The term "buffer solution" is intended to mean an aqueous solution containing an acid-based mixture for the purpose of stabilizing the pH. Examples of buffers are magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic compatible substances used in pharmaceutical formulations.

[0025] The term "diluent" is intended to mean an aqueous or non-aqueous solution having the purpose of diluting a compound in a pharmaceutical preparation. The diluent may be one or more of physiological saline, water, polyethylene glycol, propylene glycol, or ethanol.

[0026] Excipients may be one or more of carbohydrates, surfactants, polymers, lipids, and minerals. Examples of carbohydrates include lactose, sucrose, mannitol, and cyclodextrin, which are added to the composition, for example, to facilitate lyophilization. Examples of polymers include starch, cellulose ether, carboxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, ethyl hydroxyethyl cellulose, alginate, carrageenan, hyaluronic acid and its derivatives, polyacrylic acid, polysulfonate, polyethylene glycol / polyethylene oxide, polyethylene oxide / polypropylene oxide copolymer, polyvinyl alcohol / polyvinyl acetate of different degrees of hydrolysis, and polyvinyl pyrrolidone, all of different molecular weights, which are added to the composition, for example, for viscosity adjustment, for obtaining bioadhesion, or for protecting lipids from chemical and proteolytic degradation. Examples of lipids include fatty acids, phospholipids, mono-, di-, and triglycerides, ceramides, sphingolipids and glycolipids, all of different acyl chain lengths and degrees of saturation, egg lecithin, soy lecithin, hydrogenated egg and soy lecithin, which are added to the composition for the same reasons as polymers. Examples of minerals include talc, magnesium oxide, zinc oxide, and titanium oxide, which are added to the composition to obtain benefits, such as reduction of liquid accumulation, or beneficial pigment properties.

[0027] Examples of suitable aqueous and non-aqueous carriers that may be used in pharmaceutical compositions include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. For example, by using coating materials such as lecithin, maintaining the required particle size in the case of dispersions, and using surfactants, appropriate fluidity can be maintained.

[0028] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of microbial activity may be ensured by including various antibacterial and antifungal agents such as parabens, chlorobutanol, phenolsorbic acid, etc. It is also desirable to include isotonic agents such as sugars, sodium chloride, etc. in the composition.

[0029] When the product is a tablet, it may contain at least one additive selected from the group consisting of binders, lubricants, emulsifying agents, fillers, surfactants (such as polysorbate 80 and sodium lauryl sulfate), flavoring agents, aromas (examples of flavor-providing components) (such as orange, lemon, bergamot, grapefruit, banana, apricot, and strawberry), and natural or synthetic colorants, vitamins, sweeteners (examples of flavor-providing components) (acesulfame potassium, sodium saccharin, aspartame, stevia, and sucralose), nutritional additives (such as antioxidants, peptides), and mixtures thereof.

[0030] Furthermore, the tablets may contain various lubricants suitable for use in compositions containing water-dispersible, water-soluble, water-insoluble lubricants, and combinations thereof. Examples of useful water-soluble lubricants include sodium benzoate, polyethylene glycol, L-leucine, adipic acid, and combinations thereof.

[0031] The tablets may also contain water-insoluble lubricants including, for example, stearates (such as magnesium stearate, calcium stearate, and zinc stearate), oils (such as mineral oil, hydrogenated and partially hydrogenated vegetable oils, and cottonseed oil), and combinations thereof.

[0032] In light of the present invention, pre-treatment is understood to be a treatment that is carried out prior to another treatment to promote the effect of the other treatment or reduce side effects. More specifically, pre-treatment of recurrent cancer involves sensitizing cancer cells prior to their treatment by another treatment, thereby promoting the effect of the other treatment. The pre-treatment itself typically has only a limited effect on cancer cells. Rather, it is the combination of pre-treatment and another treatment that provides an effective treatment for recurrent cancer cells.

[0033] Recurrent cancer is preferably selected from the group consisting of sarcoma, malignant melanoma, skin cancer, estrogen receptor-dependent and independent breast cancer, ovarian cancer, prostate cancer, kidney cancer, colon and rectal cancer, pancreatic cancer, head and neck cancer, small cell and non-small cell lung cancer, and cancers of blood cells.

[0034] The treatment of recurrent cancer should be selected based on the type of cancer the subject is suffering from. The treatment particularly includes chemotherapy and / or immunotherapy. Chemotherapy may include the administration of CHOP, which is a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone, and may be administered in amounts of 750 + / - 10% mg / m 2 of cyclophosphamide, 50 + / - 10% mg / m 2 of doxorubicin, 1.4 + / - 10% mg / m 2 of vincristine, and 50 + / - 10% mg / m 2 of prednisone, where m 2 refers to the human body surface area.

[0035] Immunotherapy may include the administration of an antibody, monoclonal antibody, or a functional fragment thereof, such as rituximab, ofatumumab, GA101, tositumomab, ibritumomab, ocrelizumab, belimumab, epratuzumab, FTBA05, AME-133V, or R603. All of the above-mentioned antibodies bind to CD20 present on B cells. The antibody may be administered in an amount of 375 + / - 10% mg / m 2 and may be administered in an amount of.

[0036] Treatment is typically performed using a therapeutic agent. The therapeutic agent may include one or more chemotherapeutic substances and / or immunotherapeutic substances as described above. Other examples of treatment and therapeutic agents are surgery, radiation, and gene therapy.

[0037] The HDAC inhibitor is administered to a subject. The subject is preferably a mammal, more preferably a human.

[0038] The HDAC inhibitor should be administered in a pharmaceutically effective dose. By "pharmaceutically effective dose" is meant a dose sufficient to produce a desired effect with respect to the condition in which it is administered. The exact dose depends on the method of administration, the nature and severity of the disorder, and the general health, sex, age, and weight of the patient. The total amount of the HDAC inhibitor should be selected to provide a desired concentration of the specific HDAC inhibitor or a plasma concentration of the HDAC inhibitor in the patient that is above that.

[0039] The desired plasma concentration of valproic acid is predicted to be in the range of 500 - 2500 μM, preferably 500 - 1500 μM, such as 600 - 1000 μM, during pretreatment.

[0040] The HDAC inhibitor may be administered once a day, for example, at a morning time of about 5 - 8 am. However, the administration of the HDAC inhibitor may be up to once, twice, three times, four times, or five times a day. The administration of the HDAC inhibitor may be at least 24 - 72 hours before treatment, such as 30 - 60, 40 - 50, or 48 hours before treatment.

[0041] Thus, valproic acid may be orally or intravenously administered in the range of about 500 mg to about 15,000 mg per day, such as about 4,000 mg to about 15,000 mg per day, such as about 400 mg to about 3,000 mg per day. For example, the oral dosage can be about 800, about 1,600, about 2,400, about 3,000, about 6,000, about 9,000, about 15,000 mg per day. This amount may be administered in a single dose or in multiple doses. Preferably, the HDAC inhibitor is administered based on the human body weight (to obtain the desired blood concentration), but the human body surface area can also be included. The human body surface area is preferably calculated using the following formula (Dubois & Dubois): 0.20247 × height (m) 0.725 × weight (kg) 0.425 and is calculated using

[0042] Preferably, the recurrent cancer is selected from the group consisting of relapsed diffuse large B-cell lymphoma (DLBCL), relapsed follicular lymphoma, relapsed chronic lymphocytic leukemia, and relapsed Hodgkin lymphoma. More preferably, the recurrent cancer is diffuse large B-cell lymphoma (DLBCL).

[0043] Preferably, the treatment of the recurrent cancer includes the administration of one or more therapeutic agents selected from the list consisting of bendamustine, gemcitabine, and carboplatin.

[0044] Bendamustine (1H-benzimidazole-2-butyric acid, 5-[bis(2-chloroethyl)amino]-1-methyl-, CAS number 16506-27-7) is sold under the brand name Treanda in particular. It is a chemotherapy drug used for the treatment of chronic lymphocytic leukemia (CLL), multiple myeloma, and non-Hodgkin lymphoma. It is administered by injection.

[0045] Gemcitabine (cytidine, 2'-deoxy-2',2'-difluoro-, CAS No. 95058-81-4) is a chemotherapeutic agent. It treats cancers including testicular cancer, breast cancer, ovarian cancer, non-small cell lung cancer, pancreatic cancer, and bladder cancer. It is administered by intravenous injection.

[0046] Carboplatin (platinum, diamine[1,1-cyclobutanedicarboxylato(2-)-κO,κO’’]-, (SP-4-2)-, CAS No. 41575-94-4) is a chemotherapeutic agent used particularly to treat ovarian cancer, lung cancer, head and neck cancer, brain cancer, and neuroblastoma. It is used by injection.

[0047] Typically, a recurrent cancer is a recurrence of a primary or prior cancer. Recurrence means that the cancer occurs again after a period when the cancer could not be detected. The recurrent cancer may be located in the same location as the primary cancer (tumor) or may appear at a different location in the subject's body. Primary cancer and prior cancer refer to the occurrence of a previous, typically first, cancer in the subject.

[0048] Preferably, the primary or prior cancer is selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia, and Hodgkin lymphoma. More preferably, the primary or prior cancer is diffuse large B-cell lymphoma (DLBCL).

[0049] Preferably, the primary or prior cancer has been treated with chemotherapy and / or immunotherapy. Chemotherapy includes the administration of one or more chemotherapeutic agents. Immunotherapy is a treatment that uses certain parts of the subject's immune system to fight cancer. Immunotherapy may include the administration of immunotherapeutic agents such as checkpoint inhibitors, cytokines, immunomodulators, cancer vaccines, monoclonal antibodies, and oncolytic viruses, and treatments such as chimeric antigen receptor (CAR) T-cell therapy.

[0050] Preferably, the chemotherapy includes a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP). Preferably, the immunotherapy includes a CD20 antibody, preferably rituximab.

[0051] Preferably, the recurrent cancer is relapsed diffuse large B-cell lymphoma (DLBCL), the primary or previous cancer is diffuse large B-cell lymphoma (DLBCL), and the subject suffering from the recurrent cancer has been treated for the primary or previous cancer using a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), or a combination of a CD20 antibody, preferably rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP).

[0052] Preferably, the steroid is also administered to the subject suffering from the recurrent cancer as part of the pre-treatment of the recurrent cancer. The steroid may be administered before, after, or simultaneously with the HDAC inhibitor. The steroid may be administered in its own pharmaceutical composition or may be included in a pharmaceutical composition containing the HDAC inhibitor. The steroid may increase the effect of the pre-treatment.

[0053] Preferably, the steroid is selected from the group consisting of prednisone, prednisolone, dexamethasone, or betamethasone, and the steroid is preferably selected from the group consisting of prednisone and prednisolone. Prednisone or prednisolone may be administered in an amount of 20-200 mg per day, such as 50-200, 100-150, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg per day. This amount may be administered in a single dose or in multiple doses.

[0054] Betamethasone may be administered in an amount of 4 to 32 mg per day, such as 10 to 25, 10 to 20, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 mg per day. This amount may be administered as a single dose or as multiple doses.

[0055] Dexamethasone may be administered in an amount of 10 to 80 mg per day, such as 20 to 70, 10, 20, 30, 40, 50, 60, 70, or 80 mg per day. This amount may be administered as a single dose or as multiple doses.

[0056] A fourth aspect of the present invention is a histone deacetylase (HDAC) inhibitor and a therapeutic agent for use in combination therapy for recurrent cancer, wherein the HDAC inhibitor is selected from the group consisting of valproic acid, valproic acid semisodium, valproic acid sodium, valproic acid magnesium, or mixtures thereof, the therapeutic agent is selected from the list consisting of bendamustine, gemcitabine, and carboplatin, and the HDAC inhibitor is administered to a subject suffering from recurrent cancer prior to the therapeutic agent being administered to the subject suffering from recurrent cancer. It relates to a histone deacetylase (HDAC) inhibitor and a therapeutic agent.

[0057] A corresponding fifth aspect of the present invention is a method of combination therapy for recurrent cancer, a step of pre-treating recurrent cancer by administering to a subject suffering from recurrent cancer a histone deacetylase HDAC inhibitor selected from the group consisting of valproic acid, valproic acid semisodium, valproic acid sodium, valproic acid magnesium, or mixtures thereof, and a step of treating recurrent cancer by administering to a subject suffering from recurrent cancer a therapeutic agent selected from the list consisting of bendamustine, gemcitabine, and carboplatin and relates to a method.

[0058] A sixth aspect of the present invention is a kit for pre-treating and treating recurrent cancer, comprising: one or more doses of an HDAC inhibitor selected from the group consisting of valproic acid, sodium valproate semihydrate, sodium valproate, magnesium valproate, or a mixture thereof; one or more doses of a therapeutic agent selected from the list consisting of bendamustine, gemcitabine, and carboplatin The present invention relates to a kit comprising the same.

[0059] The doses are preferably supplied in suitable containers and / or suitable packaging, and the kit preferably further comprises instructions for use on when and how to administer the doses. Thus, the kit may include, for example, one or more blisters for holding the doses. Preferably, the kit includes a plurality of foldable blisters sealed by a breakable seal sheet, each of the blisters containing a blister pack holding one or more doses. The kit may further include additional components, such as a suitable solution for dilution (e.g., physiological saline, glucose solution, etc.), reagents (e.g., for adjusting the pH), and equipment for assembly and use (e.g., bags, tubes, syringes, needles, transfer sets).

[0060] Preferably, the kit further comprises one or more doses of a steroid selected from the group consisting of prednisone, prednisolone, dexamethasone, or betamethasone, and the steroid is preferably selected from the group consisting of prednisone and prednisolone.

[0061] Preferably, the kit comprises one or more doses of an HDAC inhibitor selected from the group consisting of valproic acid, sodium valproate semihydrate, sodium valproate, magnesium valproate, or a mixture thereof, and one or more doses of a therapeutic agent selected from the list consisting of bendamustine, gemcitabine, and carboplatin, and optionally A steroid selected from the group consisting of prednisone, prednisolone, dexamethasone, or betamethasone, preferably selected from the group consisting of prednisone and prednisolone, one or more doses of the steroid comprising.

[0062] In other words, it is preferable that the kit does not contain any other HDAC inhibitors, therapeutic agents, and steroids other than the HDAC inhibitors, therapeutic agents, and steroids listed above.

Example

[0063] Example 1: Valproic acid enhances the effects of bendamustine, gemcitabine, and carboplatin on the growth of B-cell lymphoma In this example, the combined effects of valproic acid and three different compounds, and single treatments on the growth of two cell lines were measured by the WST 1 assay.

[0064] Cell lines used: SU DHL 8: Human B-cell lymphoma, and WSU NHL: Human B-cell lymphoma The WST 1 colorimetric assay involved supplying suspended cells (5000 cells / well) into the wells of a well plate. The tetrazolium salt (WST 1) was added to the cells, thereby coloring the liquid in the well red. Viable cells reduce the tetrazolium salt via an intermediate electron acceptor to form a formazan product, and the liquid in the cell turns yellow. The degree of color change reflects cell growth and is read as the optical absorption at 420 nm. It was found that both cell lines could tolerate a concentration of dimethyl sulfoxide (DMSO) up to 1%, and this DMSO concentration was used as a negative control. Additionally, 10% DMSO was used as a positive control.

[0065] In the pretreatment step, the cells of each cell line were incubated with 1 mM valproic acid for 48 hours, and then a treatment step with each compound for 24 or 48 hours was performed as shown in Table 1 below.

[0066]

Table 1

[0067] Figure 1A shows the results of pre-treatment of cells from the WSU-NHL cell line and subsequent 24-hour treatment with each compound for cell proliferation. The Y-axis represents the percentage of proliferation of treated cells normalized to the values obtained from cells treated with 1% DMSO in the absence or presence of valproic acid (VPA). As seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of 24-hour treatment with each compound.

[0068] Figure 1B shows the results of pre-treatment of cells from the WSU-NHL cell line and subsequent 24-hour treatment with each compound for cell proliferation. Here, the Y-axis represents the percentage of proliferation of treated cells normalized to the values obtained from untreated control cells. As seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of 24-hour treatment with each compound. In particular, significant effects were seen at the 5 nM level of gemcitabine and 5 μM of bendamustine.

[0069] Figure 2A shows the results of pre-treatment of cells from the WSU-NHL cell line and subsequent 48-hour treatment with each compound for cell proliferation. The Y-axis represents the percentage of proliferation of treated cells normalized to the values obtained from cells treated with 1 / DMSO in the absence or presence of valproic acid (VPA). As seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of 48-hour treatment with each compound.

[0070] Figure 2B shows the results of pre-treatment of cells from the WSU-NHL cell line and subsequent 48-hour treatment with each compound for cell proliferation. Here, the Y-axis represents the percentage of proliferation of the treated cells normalized to the values obtained in untreated control cells. As seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of the 48-hour treatment with each compound. In particular, significant effects were seen at the 5 nM level of gemcitabine and at 5 μM of bendamustine and carboplatin.

[0071] Figure 3A shows the results of pre-treatment of cells from the SU-DHL cell line and subsequent 24-hour treatment with each compound for cell proliferation. The Y-axis represents the percentage of proliferation of the treated cells normalized to the values obtained in cells treated with 1% DMSO in the absence or presence of valproic acid (VPA). As seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of the 24-hour treatment with each compound.

[0072] Figure 3B shows the results of pre-treatment of cells from the SU-DHL cell line and subsequent 24-hour treatment with each compound for cell proliferation. Here, the Y-axis represents the percentage of proliferation of the treated cells normalized to the values obtained in untreated control cells. As seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of the 24-hour treatment with each compound.

[0073] Figure 4A shows the results of pre-treatment of cells from the SU-DHL cell line and subsequent 48-hour treatment with each compound for cell proliferation. The Y-axis represents the percentage of proliferation of the treated cells normalized to the values obtained in cells treated with 1 / DMSO in the absence or presence of valproic acid (VPA). As seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of the 48-hour treatment with each compound. The combination of VPA with 25 nM of gemcitabine and 25 μM of bendamustine killed all cancer cells.

[0074] Figure 4B shows the results of pre-treatment of cells from the SU-DHL cell line and subsequent 48-hour treatment with each compound for cell proliferation. Here, the Y-axis represents the percentage of proliferation of treated cells normalized to the values obtained in untreated control cells. As can be seen in the graph, 48-hour pre-treatment with valproic acid increased the effect of 48-hour treatment with each compound. The combination of VPA and carboplatin (at both 5 μM and 25 μM) leaves approximately 10 viable cells.

[0075] As can be seen from the above examples and drawings, valproic acid (VPA) is effective as a pre-treatment for subsequent treatment using the therapeutic agents gemcitabine, bendamustine, and carboplatin, for which the agent is used to treat recurrent cancer.

Claims

1. A histone deacetylase (HDAC) inhibitor for use in pretreatment of recurrent cancer prior to treatment of the recurrent cancer, the histone deacetylase (HDAC) inhibitor selected from the group consisting of valproic acid, semisodium valproic acid, sodium valproic acid, magnesium valproic acid, or a mixture thereof, for administration to a subject suffering from the recurrent cancer.

2. A histone deacetylase (HDAC) inhibitor for use according to claim 1, wherein the recurrent cancer is selected from the group consisting of recurrent diffuse large B-cell lymphoma (DLBCL), recurrent follicular lymphoma, recurrent chronic lymphocytic leukemia, and recurrent Hodgkin lymphoma.

3. A histone deacetylase (HDAC) inhibitor for use according to claim 1 or 2, wherein the treatment of the recurrent cancer comprises the administration of one or more therapeutic agents selected from the list consisting of bendamustine, gemcitabine, and carboplatin.

4. A histone deacetylase (HDAC) inhibitor for use according to claim 1 or 2, wherein the recurrent cancer is a recurrence of a primary or previously existing cancer.

5. A histone deacetylase (HDAC) inhibitor for use according to claim 4, wherein the primary or past cancer is selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia, and Hodgkin lymphoma.

6. A histone deacetylase (HDAC) inhibitor for use according to claim 1 or 2, wherein the primary or past cancer has been treated with chemotherapy and / or immunotherapy.

7. A histone deacetylase (HDAC) inhibitor for use according to claim 1 or 2, wherein the recurrent cancer is recurrent diffuse large B-cell lymphoma (DLBCL), the primary or past cancer is diffuse large B-cell lymphoma (DLBCL), and the subject suffering from the recurrent cancer has previously been treated for the primary or past cancer with a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), or a CD20 antibody, preferably a combination of rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP).

8. A histone deacetylase (HDAC) inhibitor for use according to claim 1 or 2, wherein the steroid is also administered to the subject having the recurrent cancer as part of the prior treatment for the recurrent cancer.

9. The histone deacetylase (HDAC) inhibitor for use according to claim 8, wherein the steroid is selected from the group consisting of prednisone, prednisolone, dexamethasone, or betamethasone, and preferably the steroid is selected from the group consisting of prednisone and prednisolone.

10. A kit for pretreatment and treatment of recurrent cancer, One or more doses of an HDAC inhibitor selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, magnesium valproate, or mixtures thereof. A kit containing one or more doses of a therapeutic agent selected from a list consisting of bendamustine, gemcitabine, and carboplatin.

11. The kit according to claim 10, further comprising one or more doses of a steroid selected from the group consisting of prednisone, prednisolone, dexamethasone, or betamethasone, wherein the steroid is preferably selected from the group consisting of prednisone and prednisolone.

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