Combination therapy containing panobinostat for the treatment of cholangiocarcinoma

Abstract

The present invention relates to compositions and methods for treating cholangiocarcinoma, and in particular to combination therapy comprising a panobinostat composition in combination with other cytotoxic agents (e.g., agents that promote the action of panobinostat) for treating cholangiocarcinoma. Also provided are pharmaceutical compositions comprising panobinostat and other cytotoxic agents.

Description

Technical Field

[0001] The present invention relates to compositions and methods for treating cholangiocarcinoma. More specifically, the present invention relates to combination therapy comprising a panobinostat composition in combination with other cytotoxic agents (e.g., agents that enhance the effect of panobinostat) for treating cholangiocarcinoma, and methods for treating cholangiocarcinoma by co-administering panobinostat with other cytotoxic agents (e.g., agents that promote the effect of panobinostat). Background Art

[0002] There are more than 100 forms of cancer that originate from specific cell types that are not associated with organs or tissues. The National Cancer Institute (NCI) lists the major types of cancer (https: / / www.cancer.gov / types), each of which can be further grouped and classified based on the expression of molecular markers, gene expression profiles, mutational load, and transforming oncogenic mutations. Breast cancer is one such example, which is further classified based on the expression of estrogen receptors, progesterone receptors, and HER2 receptors. In addition, triple-negative breast cancer does not express any of the above receptors.

[0003] As with almost all forms of cancer, the prognosis is much better if the tumor is diagnosed early in the disease's progression, and if the cancer is also grouped according to the stage it has advanced in. Different forms and stages of cancer often have different treatment options.

[0004] Cancer treatment for any given diagnosis is further divided into first-line, second-line, and third-line treatments if treatment is based on established and available treatment options. The preferred treatment options for various forms of cancer may also vary somewhat between countries.

[0005] Cholangiocarcinoma (CCA) is one of the rare primary malignancies in Europe and North America. However, it is more common in Asian countries (Boris Blechacz: Cholangiocarcinoma: Current Knowledge and New Developments in Gut Liver., January 2017; Vol. 11 No. 1: pp. 13-26).

[0006] In cholangiocarcinoma, the cancer cells originate from either the intrahepatic or extrahepatic bile ducts. Therefore, cholangiocarcinoma can be divided into intrahepatic and extrahepatic cholangiocarcinoma. Extrahepatic cholangiocarcinoma accounts for 60%-80% of cholangiocarcinomas and can be subdivided into hilar cholangiocarcinoma and distal cholangiocarcinoma. In Norway, the main treatment option for cholangiocarcinoma is surgery. However, 70%-80% of extrahepatic cholangiocarcinomas are not amenable to curative resection. Radiotherapy may be another valuable treatment option. If patients have metastatic cholangiocarcinoma, medical treatment is usually gemcitabine with oxaliplatin, capecitabine, or cisplatin.

[0007] In recent years, various clinical studies on the treatment of cholangiocarcinoma with drugs and drug combinations have been reported in scientific literature and databases. These treatment studies include targeted therapies such as monoclonal antibodies ("mAbs"), kinase inhibitors ("Nibs"), and other drugs.

[0008] For example, WO2017 / 202806 relates to peptides and combinations of peptides for use in immunotherapy against gallbladder cancer and cholangiocarcinoma, as well as other cancers.

[0009] WO2017 / 037299 provides a method for treating cholangiocarcinoma by administering a therapeutically effective amount of vanitinib.

[0010] WO2008 / 023947 describes a pharmaceutical composition for inhibiting the growth or metastasis of cholangiocarcinoma, comprising an inhibitor of L1CAM activity or expression, and a therapeutic method using the composition.

[0011] However, despite the development of new therapies, cholangiocarcinoma is still considered a devastating malignancy with fatal complications and low response and resistance to chemotherapy.

[0012] The prognosis of patients with cholangiocarcinoma is usually very poor and drug therapy has limited clinical value in cholangiocarcinoma. The five-year survival rate of cholangiocarcinoma is less than 5%, and the five-year survival rate of patients who cannot undergo surgery is 0%. The average survival time is 12 months. Therefore, the medical community urgently needs improved therapies.

[0013] Brief description of the invention

[0014] In the work to realize the present invention, the inventors selected more than 380 known anti-cancer related drug substances (e.g., cytotoxic agents) for extensive evaluation of their effects on several cholangiocarcinoma cell lines in both single and combined therapy. After several rounds of selection based on the known properties of the substances (such as efficacy at low doses, benign side effects, and known mechanisms of action) combined with their activity on cholangiocarcinoma cell lines, the inventors found that panobinostat was particularly effective against both intrahepatic and extrahepatic cholangiocarcinoma cell lines. This was particularly surprising considering that only a portion of the tested drugs and drug combinations were active in the cell assay.

[0015] Furthermore, the inventors determined that some selected drug substances (eg, cytotoxic agents) can enhance the anticancer activity of panobinostat against one or more cholangiocarcinoma cell lines.

[0016] Therefore, in its broadest sense, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and administering to the subject a therapeutically effective amount of a cytotoxic agent that promotes (i.e., enhances) the therapeutic effect of panobinostat or a pharmaceutically acceptable salt thereof, wherein the cytotoxic agent is administered separately, simultaneously or sequentially with the therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0017] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent that promotes (ie, enhances) the therapeutic effect of panobinostat or a pharmaceutically acceptable salt thereof, for use in treating cholangiocarcinoma in a subject.

[0018] In another embodiment, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof for use in treating cholangiocarcinoma in a subject in combination with a cytotoxic agent that promotes (ie, enhances) the therapeutic effect of panobinostat or a pharmaceutically acceptable salt thereof.

[0019] In yet another embodiment, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with a cytotoxic agent for treating cholangiocarcinoma in the subject. In some embodiments, panobinostat or a pharmaceutically acceptable salt thereof can be formulated with a cytotoxic agent to provide a combined preparation, such as a pharmaceutical composition comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent.

[0020] The present invention also provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for use or administration to a subject in combination with a cytotoxic agent for separate, simultaneous or sequential use to treat cholangiocarcinoma in the subject. In some embodiments, panobinostat or a pharmaceutically acceptable salt thereof can be formulated with a cytotoxic agent to provide a combined preparation, such as a pharmaceutical composition comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent. DETAILED DESCRIPTION

[0021] Panobinostat ((E)-N-hydroxy-3-[4-[[2-(2-methyl-1H-indol-3-yl)ethylamino]methyl]phenyl]prop-2-enamide) is an enzyme inhibitor of histone deacetylase (HDAC) having the following structure. Panobinostat is available from Novartis. Alternatively, panobinostat may be prepared as described in WO 02 / 22577, which is incorporated herein by reference. Panobinostat referred to herein includes salts thereof.

[0022]

[0023] Pharmaceutically acceptable salts include pharmaceutically acceptable base addition salts and acid addition salts, for example metal salts, such as alkali metal salts and alkaline earth metal salts, ammonium salts, organic amine addition salts and amino acid addition salts, and sulfonates. Acid addition salts include inorganic acid addition salts (such as hydrochlorides, sulfates and phosphates) and organic acid addition salts (such as alkyl sulfonates, aryl sulfonates, acetates, maleates, fumarates, tartrates, citrates and lactates). Examples of metal salts are alkali metal salts such as lithium salts, sodium salts and potassium salts; alkaline earth metal salts such as magnesium salts and calcium salts; aluminum salts; and zinc salts. Examples of ammonium salts are ammonium salts and tetramethylammonium salts. Examples of organic amine addition salts are morpholinium salts and piperidine salts. Examples of amino acid addition salts are glycinates, phenylalaninates, glutamates and lysine salts. Sulfonates include mesylate, toluenesulfonate and benzenesulfonate.

[0024] Preferred salts include organic acid addition salts such as alkyl sulfonates, aryl sulfonates, acetates, maleates, fumarates, tartrates, citrates and lactates. Lactates are particularly preferred.

[0025] Unless otherwise indicated, the above listing of pharmaceutically acceptable salts applies to all drug substances described herein (eg, panobinostat and the cytotoxic agents described below).

[0026] As used herein, "pharmaceutically acceptable" refers to ingredients that are compatible with other ingredients used in the methods or uses of the present invention and are physiologically acceptable to the recipient.

[0027] "Cholecystocarcinoma" or "CCA" can be intrahepatic cholangiocarcinoma or extrahepatic cholangiocarcinoma (which can be hilar cholangiocarcinoma and distal cholangiocarcinoma). More than 90% of cholangiocarcinomas are adenocarcinomas. In some embodiments, the cholangiocarcinoma to be treated is metastatic cholangiocarcinoma. In some embodiments, the cholangiocarcinoma to be treated is intrahepatic cholangiocarcinoma. In some embodiments, the cholangiocarcinoma to be treated is extrahepatic cholangiocarcinoma.

[0028] As described in detail in the embodiments, the inventors have determined that the conjugate therapy of the present invention has different effects in various cell lines. In this respect, cell lines are derived from individual tumors and can be considered to represent different forms of cholangiocarcinoma. For example, each cell line can have one or more characteristics, such as one or more genetic markers, growth rate, cell morphology or a combination thereof, which are generally found in cholangiocarcinoma tumors. Therefore, the conjugate therapy disclosed herein that is particularly effective in inhibiting the growth of a specific cell line or killing cells of a specific cell line can be found to have special utility in treating cholangiocarcinoma tumors, and the cholangiocarcinoma tumor has one or more characteristics (such as one or more genetic markers (such as mutations), growth rate and / or cell morphology) associated with a cholangiocarcinoma cell line, such as one or more characteristics specific to a cholangiocarcinoma cell line.

[0029] For example, the EGI-1 (CVCL_1193) and TFK-1 (CVCL_2214) cell lines are derived from explants of extrahepatic cholangiocarcinoma tumors in male subjects (Shimizu et al., Int. J. Cancer, Vol. 52: pp. 252-260 (1992), incorporated herein by reference). Therefore, in some embodiments, the combination therapy disclosed herein can be used to treat a subject with a cholangiocarcinoma tumor (e.g., an extrahepatic cholangiocarcinoma tumor) that has one or more characteristics specific to the EGI-1 cell line and / or the TFK-1 cell line, such as one or more genetic markers, growth rate and / or cell morphology.

[0030] The CC-SW-1 cell line is derived from an explant of an intrahepatic cholangiocarcinoma tumor in a female subject. Thus, in some embodiments, the combination therapies disclosed herein can be used to treat a subject having a cholangiocarcinoma tumor (e.g., an intrahepatic cholangiocarcinoma tumor) that has one or more characteristics specific to the CC-SW-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0031] The HuCC-T1 cell line is derived from ascites of a male subject with metastatic intrahepatic cholangiocarcinoma tumors. Thus, in some embodiments, the combination therapies disclosed herein can be used to treat subjects with cholangiocarcinoma tumors (e.g., intrahepatic cholangiocarcinoma tumors) (e.g., subjects with metastatic intrahepatic cholangiocarcinoma) that have one or more characteristics specific to the HuCC-T1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0032] The Examples section describes which combination therapies are effective in each cell line, and therefore these therapies may be effective in treating cholangiocarcinoma tumors as defined above. In representative examples, Examples 26 and 27 show that trametinib and doxorubicin are particularly effective in promoting the effects of panobinostat in the CC-SW-1 cell line. Therefore, in some embodiments, the present invention provides a combination therapy of panobinostat with trametinib or doxorubicin (as defined herein, e.g., including salts thereof, etc.) for treating a subject having a cholangiocarcinoma tumor (e.g., an intrahepatic cholangiocarcinoma tumor) having one or more characteristics specific to the CC-SW-1 cell line, e.g., one or more genetic markers, growth rate, and / or cell morphology.

[0033] Because some combination therapies are effective against more than one cell line, e.g., trametinib can enhance the effect of panobinostat in CC-SW-1, EGI-1, HuCC-T1, and TFK-1 cell lines, the combination therapies may find utility in treating subjects having cholangiocarcinoma tumors that have one or more characteristics (e.g., one or more genetic markers, growth rate, and / or cell morphology) that are specific to multiple particular cell lines, e.g., cholangiocarcinoma tumors that have characteristics that are specific to the CC-SW-1 cell line and characteristics that are specific to the HuCC-T1 cell line.

[0034] A feature or combination of features (e.g., a combination of genetic markers such as mutations) that is specific to a cholangiocarcinoma cell line refers to a feature or combination of features that is present in a cholangiocarcinoma cell line and is not present in normal (i.e., healthy) cholangiocytes and / or one or more other cholangiocarcinoma cell lines.

[0035] In this regard, Example 29 describes the genetic analysis of the above-mentioned cholangiocarcinoma cell lines and the mutations identified in each cell line. Therefore, in some embodiments, cholangiocarcinoma tumors with one or more characteristics associated with the EGI-1 cell line may have one or more mutations in genes selected from KRAS, TP53, ASXL1, PDGFRA, MYH11, E2F1, AHNAK, SAFB2, NOTCH1, PEG3, CADM3, SPI1, AR, HCAR2, PPP1R1B or a combination thereof. In some embodiments, mutations in these genes are as described in Example 29. In some embodiments, cholangiocarcinoma tumors with one or more characteristics associated with the EGI-1 cell line may have one or more mutations in genes selected from KRAS and / or TP53, particularly Gly12Asp in KRAS and / or Arg273His in TP53.

[0036] In some embodiments, cholangiocarcinoma tumors having one or more characteristics associated with the TFK-1 cell line may have one or more mutations in a gene selected from BAP1, PBRM1, IKZF3, PAWR, FGFR3, STIL, SEMA3F, PCM1, FGF5, WHSC1, TP53 (e.g., Trp91Ter, 272G>A), or a combination thereof. In some embodiments, the mutations in these genes are as described in Example 29.

[0037] In some embodiments, cholangiocarcinoma tumors having one or more characteristics associated with the HuCC-T1 cell line may have one or more mutations in genes selected from KRAS, TP53, FBXW7, LETMD1, SETD2, KDM5A, MYO18B, RB1, DNAJA3, CDT1, ZFP36L2, MAF, GMPS, NPAS2, CNTNAP2, MSH6 (e.g., Lys1358fs*2, coding sequence 4071_4072insGATT), or a combination thereof. In some embodiments, mutations in these genes are as described in Example 29. In some embodiments, cholangiocarcinoma tumors having one or more characteristics associated with the HuCC-T1 cell line may have one or more mutations in genes selected from KRAS and / or TP53, particularly Gly12Asp in KRAS and / or Arg175His in TP53.

[0038] In some embodiments, cholangiocarcinoma tumors having one or more characteristics associated with the CC-SW-1 cell line may have one or more mutations in a gene selected from PDGFRA, CCAR2, RECK, ZNF292, PYHIN1, DSP, or a combination thereof. In some embodiments, the mutations in these genes are as described in Example 29.

[0039] As defined herein, "treatment" as used herein refers generally to any action or step (or intervention) that is beneficial to the management of a clinical condition or disorder. Thus, treatment may refer to reducing, alleviating, ameliorating, slowing down the development of one or more symptoms of cholangiocarcinoma (CCA) being treated, or eliminating these symptoms, or improving the clinical status of the subject in any way, relative to the symptoms before treatment. Treatment may include any clinical step or intervention that contributes to or becomes part of a treatment procedure or regimen. In particular, the treatment may include reducing the tumor size or volume of the cholangiocarcinoma being treated.

[0040] Treatment may include delaying, limiting, reducing or preventing the onset of one or more symptoms in bile duct cancer, for example relative to the bile duct cancer or symptoms before treatment. Thus, treatment explicitly includes absolute prevention of the onset or development of symptoms in bile duct cancer and any delay in the development of bile duct cancer or symptoms, or reducing or limiting the development or progression of bile duct cancer or symptoms.

[0041] Thus, treatment according to the invention includes killing, inhibiting or slowing the growth of cholangiocarcinoma cells or the increase in the size of a body or population of cholangiocarcinoma cells (e.g., in a tissue, in a tumor, or in a growth), reducing the number of cholangiocarcinoma cells or preventing the spread of cholangiocarcinoma cells (e.g., to another anatomical site), reducing the size of cell growths, etc. The term "treating" does not necessarily mean cure or complete removal or elimination of cholangiocarcinoma cell growth (or growth of cholangiocarcinoma cells).

[0042] A "subject" or "patient" is an animal (ie, any human or non-human animal), preferably a mammal, most preferably a human.

[0043] The therapeutic agents or drug substances described herein (eg, panobinostat, cytotoxic agents) can be administered to a subject using any suitable means, and the route of administration will depend on the therapeutic agent. In some embodiments, the therapeutic agent is administered systemically.

[0044] "Systemic administration" includes any form of non-local administration, in which the agent is administered to the body at a site other than directly adjacent to the cholangiocarcinoma or the local vicinity of the cholangiocarcinoma so that the systemic administration of the agent is received. Conveniently, systemic administration can be by enteral delivery (e.g., oral) or parenteral delivery (e.g., intravenous administration, intramuscular administration, or subcutaneous administration).

[0045] Panobinostat can be administered in any suitable pharmaceutical form. For example, panobinostat can be provided as a pharmaceutical composition comprising panobinostat or a salt thereof and a pharmacologically (or pharmaceutically) acceptable excipient.

[0046] Excipients may include any excipient known in the art, for example any carrier or diluent or any other ingredient or agent, such as a buffer, antioxidant, chelating agent, binder, coating, disintegrant, filler, flavoring agent, coloring agent, glidant, lubricant, preservative, adsorbent and / or sweetener, etc.

[0047] Excipients can be selected from, for example, lactic acid, glucose, sodium pyrosulfite, benzyl alcohol, polyethylene glycol, propylene glycol, microcrystalline cellulose, lactose, starch, chitosan, pregelatinized starch, calcium carbonate, calcium sulfate, cellulose, dextrates, dextrin, dextrose, calcium hydrogen phosphate dihydrate, tricalcium phosphate, gelatin, magnesium carbonate, magnesium oxide, magnesium stearate, maltodextrin, mannitol, powdered cellulose, pregelatinized starch, sodium chloride, sorbitol, propylene glycol and / or talc. Excipients also typically include colored substances such as titanium dioxide and various iron oxides.

[0048] Unless otherwise indicated, the above-listed list of excipients applies to all drug substances described herein (eg, panobinostat and cytotoxic agents described below).

[0049] The pharmaceutical composition described herein can be provided in any form known in the art, for example, as a tablet, capsule, coated tablet, liquid, suspension, pill, sachet, implant, powder, pellet, emulsion, lyophilizate, effervescent or any mixture thereof. It can be provided, for example, as a gastric juice resistant formulation and / or in a sustained-action form.

[0050] In a preferred embodiment, panobinostat (eg, a pharmaceutical composition comprising panobinostat or a salt thereof) is formulated for oral administration. In other words, in the methods and uses of the present invention, panobinostat is orally administered to a subject.

[0051] The most preferred dosage form of panobinostat for treating cholangiocarcinoma is in the form of tablets or capsules. The tablets may be coated tablets.

[0052] One of the even most preferred dosage forms for treating cholangiocarcinoma is in capsule form.

[0053] Panobinostat or a salt thereof may be administered in any suitable dosage range using any suitable dosage regimen. A skilled person will be aware of suitable dosage ranges for Panobinostat. In one embodiment, Panobinostat or a salt thereof is present in a pharmaceutical composition and administered to a subject in its typical dosage range. This may be considered a therapeutically effective amount of Panobinostat.

[0054] As described above, panobinostat is used in combination therapy with another therapeutic agent (e.g., a cytotoxic agent that promotes the effect of panobinostat). Therefore, in some embodiments, panobinostat can be administered at a dosage range lower than its typical dosage range. However, in the case of a lower dose of panobinostat used in a combination therapy, panobinostat will have the same or comparable therapeutic effect as a higher dose of panobinostat itself. Therefore, in some embodiments, the present invention thus makes it possible to treat subjects with low or below average tolerance to panobinostat, such as the elderly, infants or young children, or people who are debilitated, for example, due to illness, malnutrition, etc.

[0055] In a representative embodiment, the clinical dose of panobinostat for treating cholangiocarcinoma is about 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week, for example 2-6 times, 2-5 times or 2-4 times a week. In a preferred embodiment, the clinical dose is a single dose formulation, such as a tablet or capsule.

[0056] As described above and detailed in the Examples, the inventors have determined that the effects of panobinostat on cholangiocarcinoma can be enhanced when used in combination with various other cytotoxic agents (eg, anticancer agents).

[0057] Thus, the present invention relates to therapeutic regimens for treating cholangiocarcinoma wherein panobinostat is used in combination with another cytotoxic agent (eg, an anticancer drug).

[0058] Thus, the additional cytotoxic agents described herein (e.g., anticancer agents) can be used to provide a sensitizing effect, in other words, to enhance (or increase, augment, or promote) the effect of panobinostat (e.g., in the treatment of cholangiocarcinoma), or to render a subject (or more specifically, a cholangiocarcinoma cell or tumor present in a subject) more sensitive to the effect of panobinostat. Thus, in some embodiments, panobinostat can be considered a primary drug (therapeutic agent) and the additional cytotoxic agent can be considered a secondary drug (therapeutic agent).

[0059] The terms "primary drug" and "primary therapeutic agent" refer to a drug that is administered at a higher relative dose than a "secondary drug" or "secondary therapeutic agent". For example, the primary drug is administered at or near its maximum tolerated dose (e.g., at least 70%, 80%, or 90%, such as 100% of the maximum tolerated dose), and the secondary drug is administered at a dose significantly lower than its maximum tolerated dose (e.g., less than 70%, 60%, or 50% of the maximum tolerated dose). For example, the secondary drug may be administered at or near the IC20 dose. Since different drugs have different dosage ranges, it is clear that a secondary drug may be administered at a higher absolute dose than the primary drug, even if the secondary drug is administered at a dose significantly lower than its maximum tolerated dose.

[0060] The maximum tolerated dose (MTD) refers to the highest dose of a pharmacological treatment regimen that produces the desired effect without unacceptable toxicity. The skilled artisan will be aware of the maximum tolerated dose of any given cytotoxic agent disclosed herein.

[0061] In some embodiments, the additional cytotoxic agent can be any agent that reduces the IC50 value of panobinostat compared to the IC50 value of panobinostat alone. The IC50 can be determined using any suitable method, such as the in vitro method described in the Examples.

[0062] The following examples demonstrate that the IC50 of some cytotoxic agents can be reduced when used in combination with a specific dose of panobinostat. Therefore, in some embodiments, panobinostat can enhance the therapeutic efficacy of another cytotoxic agent, such as reducing the IC50 of another cytotoxic agent. In other words, in some embodiments, panobinostat can be used as (i.e., administered as) a secondary drug (e.g., at a dose significantly lower than its maximum tolerated dose), and another cytotoxic agent can be administered as a primary drug (e.g., at a dose equal to or close to its maximum tolerated dose). In some embodiments, the effect of panobinostat on the therapeutic efficacy of another cytotoxic agent (e.g., reducing the IC50 of another cytotoxic agent) can be in addition to the effect of the other cytotoxic agent on panobinostat.

[0063] In a preferred embodiment, panobinostat is used as the primary drug in the combinations disclosed herein.

[0064] The term "IC50" is a measure of the effectiveness of a substance in inhibiting a specific biological or biochemical function. Therefore, in the context of the present invention, IC50 represents the concentration of a drug (e.g., panobinostat) required for 50% inhibition (reduction) of in vitro cholangiocarcinoma cell viability. Similarly, the term "IC20" represents the concentration of a drug required for 20% inhibition (reduction) of in vitro cholangiocarcinoma cell viability. Therefore, the inhibitory concentration (IC) can be regarded as the lethal concentration (LC) or lethal dose (LD) of a substance, which terms are used to describe the administered dose in in vivo studies.

[0065] Cytotoxic agents (i.e., anticancer drugs) described herein are generally associated with adverse events in clinical use. The frequency and severity of toxicity and adverse events are generally dose-dependent. The higher the dose, the more frequent and severe the side effects. Anticancer drugs are generally used at the highest possible clinical dose (maximum tolerated dose) to maximize their efficacy. Therefore, if possible, it is clinically relevant to reduce the IC50 of anticancer drugs in cancer cells.

[0066] The ability of a cytotoxic agent to reduce the IC50 of a primary drug (e.g., panobinostat) in cholangiocarcinoma cells can be determined by measuring the change in the IC50 dose of a particular cell line to provide a ΔIC50. ΔIC50 relates to how the monotherapy curve of a given substance is affected by the combined treatment of a second compound. In some embodiments herein, a secondary drug (an additional cytotoxic agent) is added to various concentrations of a primary drug (e.g., panobinostat) at its IC20 concentration in various cell lines. When a secondary drug reduces the IC50 of a primary drug, the secondary drug can be considered to promote the effect of the primary drug. As described above, the clinical result of the combination may be that the dose of the primary drug can be reduced, thereby reducing the frequency and / or severity of side effects. Another option is to maintain the normal dose of the primary drug to improve the clinical efficacy of the drug for treating cholangiocarcinoma. In some embodiments, an additional cytotoxic agent can reduce the IC50 value of panobinostat by at least about 10%, for example, by at least about 12%, 15%, 20%, 25%, 30%, 40%, or 50%. In some embodiments, the additional cytotoxic agent can reduce the IC50 value of panobinostat by at least about 60%, 70%, 80%, 90%, or 100%.

[0067] In some embodiments, the additional cytotoxic agent is any agent that, when used in combination with panobinostat, the combination is more effective (eg, additive or synergistic) in treating cholangiocarcinoma than panobinostat alone for the same dose or the same concentration of panobinostat.

[0068] The "Combination Index" (CI) provides a quantitative assessment of the efficacy of a combination of two drug substances. For example, a combination of two drugs can act synergistically (efficacy greater than the additive efficacy of the two drugs, e.g., 2 + 2 = 5), additively (efficacy is the sum of the efficacies of the individual drugs, e.g., 2 + 2 = 4), or antagonistically (efficacy less than the sum of the efficacies of the individual drugs, e.g., 2 + 2 = 3). The CI can be calculated using the Chou-Talalay principle and CalcuSyn software (Biosoft, Ferguson, MO; see also Chou TC, Talalay P. Adv Enzyme Regul., 1984; Vol. 22: pp. 27-55; Lu Huang et al., Nature, Vol. 7, Article No.: 40752 (2017); and Ashkan Zandi et al., Middle East Journal of Cancer; January 2017; Vol. 8 No. 1: pp. 31-38, all of which are incorporated herein by reference). A CI value less than 1 indicates synergy; a CI value equal to 1 indicates additivity; a CI value greater than 1 indicates antagonism. In some embodiments, when used alone, an additional cytotoxic agent can effectively inhibit (e.g., kill) the viability of cholangiocarcinoma cells (e.g., treat cholangiocarcinoma in a subject). Thus, in some embodiments, the combination of panobinostat and an additional cytotoxic agent has an additive effect on inhibiting the viability of cholangiocarcinoma cells (e.g., treating cholangiocarcinoma in a subject), i.e., the CI value of the combination is equal to 1.

[0069] An additive interaction means that the effects of panobinostat and an additional cytotoxic agent are equal to the sum of their individual effects at the same doses, e.g., the effect is the ability of the substances to inhibit (e.g., kill) the viability of cholangiocarcinoma cells, as evaluated, for example, using the in vitro assays described in the examples.

[0070] In some embodiments, the combination of panobinostat and an additional cytotoxic agent has a synergistic effect on inhibiting (e.g., killing) the viability of cholangiocarcinoma cells (e.g., treating cholangiocarcinoma in a subject).

[0071] A synergistic interaction means that the effect of taking panobinostat and an additional cytotoxic agent together is greater than the sum of their effects when taken individually at the same doses, e.g., the effect is the ability of these substances to inhibit (e.g., kill) the viability of cholangiocarcinoma cells, as evaluated, for example, using the in vitro assays described in the examples, i.e., the CI value of the combination is less than 1, e.g., about 0.95, 0.90, 0.85, 0.80, 0.75 or less.

[0072] In some embodiments, the combination of panobinostat and an additional cytotoxic agent improves the safety factor of panobinostat for treating cholangiocarcinoma relative to the use of panobinostat alone for treating cholangiocarcinoma.

[0073] The "safety factor" is the ratio between the dose that produces toxic effects and / or severe side effects in a subject and the efficacy dose (e.g., a therapeutically effective amount). Thus, in some embodiments, the safety factor of the panobinostat combination disclosed herein is higher than the safety factor of panobinostat alone for the treatment of cholangiocarcinoma. Alternatively, in some embodiments, the additional cytotoxic agent is an agent that improves the safety factor of panobinostat.

[0074] In some embodiments, the combination of panobinostat and an additional cytotoxic agent improves the therapeutic index of panobinostat for treating cholangiocarcinoma relative to the use of panobinostat alone for treating cholangiocarcinoma.

[0075] The therapeutic index (TI) is a quantitative measure of the relative safety of a drug, which is measured as the ratio between the toxic dose (TD50) and the effective dose (ED50). ED50 is the dose that produces a given therapeutic effect in 50% of patients, and TD50 is the dose that produces a given toxic effect in 50% of patients. These values ​​can be extracted from the dose-response curve. From a clinical point of view, it is advantageous for the therapeutic index to be as high as possible. A high therapeutic index value is an indication that the drug is safe and has a low probability of serious side effects. On the other hand, if the therapeutic index is low (e.g., close to 1), the probability that a patient will produce serious side effects using a given clinical dose will be much higher. For clinically used drugs, TI will vary from drug to drug. The TI of cytotoxic drugs (e.g., anticancer drugs) is generally low, while the TI of, for example, penicillin and paracetamol is much higher.

[0076] TI can be calculated based on the ratio between IC50 in normal cells and cancer cells using in vitro data, as shown in the Examples. Thus, in some embodiments, the combination of panobinostat with an additional cytotoxic agent improves the therapeutic index, i.e., the in vitro TI, as calculated in the Examples. In some embodiments, the combined in vitro TI is at least 1.5, preferably 2.0, 2.5, 3.0 or higher, e.g., 5, 6, 7, 8, 9, 10 or higher.

[0077] The term "drug sensitivity score (DSS)" refers to a quantitative measure that characterizes a drug or drug combination in a single parameter. DDS describes the relationship between multi-parameter dose and response with a single value from 1 to 100, where higher values ​​indicate more effective treatment. DDS identifies selective drug or drug combination responses between cancer and control cells (see Yadav et al., Scientific Reports (Nature), Vol. 4, Article No.: 5193 (2014)). Therefore, in some embodiments, the combination therapy disclosed herein has a higher DSS than a monotherapy (e.g., panobinostat alone).

[0078] "Cytotoxic agent" refers to an agent that can inhibit (e.g., kill) animal cell growth, viability and / or proliferation (replication / reproduction). In some embodiments, the cytotoxic agent can inhibit (e.g., kill) cholangiocarcinoma cell (preferably, human cholangiocarcinoma cell) growth, viability and / or proliferation (replication / reproduction).

[0079] Cytotoxic agents include anti-neoplastic agents and any agent indicated for use in oncology. Thus, agents used in chemotherapy treatment regimens ("chemotherapeutic agents" or "anti-cancer" agents) are included.

[0080] Cytotoxic agents are generally classified into different categories according to their mechanism of action and all of these categories are encompassed herein. Thus, a cytotoxic agent may be, for example, an alkylating agent, a cross-linking agent, an intercalating agent, a nucleotide analog, a spindle formation inhibitor, and / or a topoisomerase I and / or II inhibitor. Other types or classes of agents include antimetabolites, plant alkaloids and terpenoids or antitumor antibiotics.

[0081] Alkylating agents modify DNA by alkylating nucleosides, which prevents proper DNA replication. Nucleotide analogs are incorporated into DNA during replication and inhibit DNA synthesis. Spindle formation inhibitors interfere with spindle formation, which stops mitosis at metaphase. Intercalators insert between DNA bases and inhibit DNA synthesis. Topoisomerase I or II inhibitors affect DNA torsion and thus interfere with DNA replication.

[0082] Suitable cytotoxic agents are known in the art, but by way of example, actinomycin D, bortezomib, BCNU (carmustine), BI 2536, bupanixiba, carboplatin, CCNU, camptothecin (CPT), cantharidin, cisplatin, combretastatin A4, CUDC-907, cyclophosphamide, cytarabine, dasatinib, dacarbazine, datoxib, dapoxetine, daunorubicin, docetaxel, doxorubicin, duvelisib, DTIC, ilisimol, epirubicin, etoposide, gefitinib, gemcitabine, idelalisib, ifosfamide, ispins, irinotecan, ionomycin, lumispivir, melphalan, methotrexate, smiloxantrone, mercaptopurine, molibresib, oxaliplatin, obaclava, paclitaxel, PARP-1 inhibitors, perifosine, PX-866, sepantronium Bromide, SB-743921, taselisib, taxane, temozolomide (TZM), teniposide, topotecan, trametinib, sulpheniramine, triptolide, erblibide, vinorelbine, vincristine, vinblastine, volacetidine, voltacil, pentazacidine, 5,6-dihydro-5-azacidine and 5-fluorouracil can be used in the combination therapy of the present invention.

[0083] In a particularly preferred embodiment, the additional cytotoxic agent is selected from bortezomib, BI 2536, carboplatin, cisplatin, combretastatin A4, datolicoxib, dapoline, dasatinib, docetaxel, doxorubicin, ilisimol, gemcitabine, ispinax, lumispivir, methotrexate, molibax, obaclava, pelitinib, SB-743921, topotecan, trametinib and triptolide.

[0084] In yet another preferred embodiment, the additional cytotoxic agent is selected from BI 2536, carboplatin, cisplatin, combretastatin A4, datolicoxib, dapoline, dasatinib, docetaxel, doxorubicin, ilisimol, ispins, lumispivir, methotrexate, molibs, obaclava, pelitinib, SB-743921, topotecan, trametinib and triptolide.

[0085] In another preferred embodiment, the additional cytotoxic agent is selected from doxorubicin, datolicoxib, SB-743921, trametinib, ilisimol, molibex, methotrexate, dapoline, topotecan, cisplatin, dasatinib, carboplatin and lumispivir.

[0086] In a further preferred embodiment, the additional cytotoxic agent is selected from carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib, datolicoxib, dapoline, ilisimol, ispins, lumispivir, molibs, obaclava, pelitinib, trametinib and triptolide, preferably selected from carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan and trametinib.

[0087] The cytotoxic agent used in combination with panobinostat may be provided in a pharmaceutical composition as defined above and may be administered as defined above and below. In some embodiments, the pharmaceutical composition comprising the cytotoxic agent may be formulated for parenteral administration. Thus, the composition may comprise pharmaceutically acceptable excipients, solvents and diluents suitable for such formulations (e.g., intravenous bolus or injection).

[0088] The skilled artisan will be aware of suitable dosage ranges for any given cytotoxic agent.In preferred embodiments, the cytotoxic agent is present in a pharmaceutical composition or administered to a subject in its typical dosage range.

[0089] However, as shown in the Examples below and described above, some cytotoxic agents are able to promote the effects of panobinostat on cholangiocarcinoma cells at low doses. Therefore, in some embodiments, the additional cytotoxic agent may be present in a pharmaceutical composition or administered to a subject at a dose range lower than the typical dose range described below. For example, in some embodiments, the additional cytotoxic agent may be present in a pharmaceutical composition or administered to a subject at a dose range of 70% or less of the typical dose range (e.g., 60%, 50%, 40% or 30% or less of the typical dose range (e.g., maximum tolerated dose)). Therefore, in some embodiments, the therapeutically effective amount of the additional cytotoxic agent is lower than the typical dose range as defined above.

[0090] In one embodiment, the combination therapy comprises administering panobinostat and bortezomib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of bortezomib or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0091] Bortezomib or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0092] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with bortezomib or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0093] In another embodiment, the present invention provides use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with bortezomib or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0094] In some embodiments, the combination therapy of panobinostat and bortezomib is used to treat intrahepatic cholangiocarcinoma.

[0095] In some embodiments, the combination therapy of panobinostat and bortezomib is used to treat a subject having a cholangiocarcinoma tumor (e.g., an intrahepatic cholangiocarcinoma tumor) having one or more characteristics specific to the CC-SW-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0096] Bortezomib ([(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]propionyl}amino)butyl]boronic acid) is a proteasome inhibitor having the structure shown below. Bortezomib is available from Janssen. The term "bortezomib" includes pharmaceutically acceptable salts, solvates and hydrates thereof.

[0097]

[0098] Stable liquid pharmaceutical compositions of bortezomib are described in WO2016 / 166653 (incorporated herein by reference) and any such compositions may be used in the methods, compositions and uses of the invention.

[0099] In some embodiments, the composition comprising bortezomib is a "ready-to-use" formulation that comprises bortezomib in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0100] In a preferred embodiment, the pharmaceutical composition comprising bortezomib is formulated for parenteral administration, such as injection or infusion.

[0101] Suitable solvent can be selected from aqueous solvent and non-aqueous solvent, for example but not limited to glycerol, ethanol, n-propanol, n-butanol, isopropanol, ethyl acetate, dimethyl carbonate, acetonitrile, methylene chloride, 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 their mixture. Preferred solvent is ethanol, glycerol and water.

[0102] The bortezomib formulation used 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.

[0103] The bortezomib preparation used 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 (diethylenetriaminepentaacetic acid), EDTA (ethylenediaminetetraacetic acid), ODDA (1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7), TTT A (1,7,13-triaza-4,10,16-trioxacyclooctadecane-N,N',N"-triacetate), DOTRP (tetraethylene glycol-1,5,9-triazacyclododecane-N,N',N"-tri(methylenephosphonic acid), EGTA (ethylene glycol-bis(P-aminoethyl ether)-tetraacetic acid), and the like.

[0104] The bortezomib formulations used 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 pyrosulfite, L-cysteine, thioglycolic acid, citric acid, tartaric acid, phosphoric acid, gluconic acid, thiodipropionic acid, etc. The most preferred antioxidant is monothioglycerol.

[0105] A most preferred aspect of the combined administration of panobinostat and bortezomib for the treatment of cholangiocarcinoma is the administration of bortezomib as a subcutaneous or intravenous injection.

[0106] The bortezomib injection used according to the present invention is preferably in the form of a water-soluble boric acid ester; the most preferred ester is mannitol borate.

[0107] The borate ester preparation, preferably mannitol ester, is usually in the form of a sterile dry powder preparation. The powder is usually freeze-dried powder. Before administration, the powder is dissolved in sterile water, usually a sterile isotonic sodium chloride aqueous solution.

[0108] The bortezomib formulation used in the present invention may optionally contain other pharmaceutically acceptable adjuvants, such as buffers, pH adjusters, preservatives, tonicity adjusters, and the like.

[0109] Unless otherwise indicated, the above-listed list of solvents, stabilizers, chelators, and antioxidants may also be used in pharmaceutical compositions comprising other cytotoxic agents described herein.

[0110] The above-mentioned bortezomib-based formulation may preferably contain mannitol and may be provided in an injection vial or in a pre-filled syringe under nitrogen protection.

[0111] A preferred embodiment of the treatment of cholangiocarcinoma using panobinostat in combination with bortezomib is to administer panobinostat orally and bortezomib as an injection.

[0112] In some embodiments, the clinical dose of panobinostat combined with bortezomib for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0113] In some embodiments, the clinical dose of bortezomib combined with panobinostat for the treatment of cholangiocarcinoma is generally 0.5 mg / m 2 Body surface area (BSA) up to 3 mg / m 2 Body surface area, at least once a week, preferably 1 mg / m 2 Body surface area (BSA) up to 2 mg / m 2 body surface area, apply at least once a week.

[0114] In a preferred aspect of the invention, administration of a combination of panobinostat and bortezomib to treat cholangiocarcinoma involves co-administration of a glucocorticoid, typically dexamethasone.

[0115] In one embodiment, the combination therapy comprises administering panobinostat and carboplatin. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of carboplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0116] Carboplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0117] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with carboplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0118] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with carboplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0119] In some embodiments, combination therapy of panobinostat and carboplatin is used to treat extrahepatic cholangiocarcinoma.

[0120] In some embodiments, the combination therapy of panobinostat and carboplatin is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0121] Carboplatin (cis-(1,1-cyclobutanedicarboxyl)diammineplatinum (II)) is a platinum-containing anticancer drug having the structure shown below. Carboplatin is widely available. The term "carboplatin" includes pharmaceutically acceptable salts, solvates and hydrates thereof.

[0122]

[0123] Liquid pharmaceutical compositions of carboplatin are well known in the art and any such compositions may be used in the methods, compositions and uses of the present invention.

[0124] In some embodiments, the composition comprising carboplatin is a "ready-to-use" formulation that comprises carboplatin in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0125] In a preferred embodiment, the pharmaceutical formulation comprising carboplatin is intended for parenteral administration.

[0126] A preferred embodiment of the treatment of cholangiocarcinoma using panobinostat in combination with carboplatin is to administer panobinostat orally and carboplatin as an injection or infusion.

[0127] In some embodiments, the clinical dose of panobinostat combined with carboplatin for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0128] In some embodiments, the clinical dose of carboplatin combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for carboplatin for other indications, such as 1 mg / m 2 BSA to 30 mg / m 2 BSA. Calvert's formula should be used to calculate the correct clinical dose.

[0129] In one embodiment, the combination therapy comprises administering panobinostat and cisplatin. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of cisplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0130] Cisplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0131] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with cisplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0132] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with cisplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0133] In some embodiments, combination therapy of panobinostat and cisplatin is used to treat extrahepatic cholangiocarcinoma.

[0134] In some embodiments, combination therapy of panobinostat and cisplatin is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line and / or the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0135] Cisplatin ((SP-4-2)-diaminedichloroplatinum (II)) is a platinum-containing anticancer drug having the structure shown below. Cisplatin is widely available, for example, from Hospira. The term "cisplatin" includes pharmaceutically acceptable salts, solvates and hydrates thereof.

[0136]

[0137] Liquid pharmaceutical compositions of cisplatin are well known in the art and any such compositions may be used in the methods, compositions and uses of the present invention.

[0138] In some embodiments, the composition comprising cisplatin is a "ready-to-use" formulation that comprises cisplatin in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0139] In a preferred embodiment, the pharmaceutical formulation comprising cisplatin is intended for parenteral administration.

[0140] A preferred embodiment of the treatment of cholangiocarcinoma using panobinostat in combination with cisplatin is to administer panobinostat orally and cisplatin as an injection or infusion.

[0141] In some embodiments, the clinical dose of panobinostat in combination with cisplatin for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0142] In some embodiments, the clinical dose of cisplatin combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for cisplatin for other indications, such as 10 mg / m 2 BSA to 50 mg / m 2 BSA, preferably 20 mg / m 2 BSA to 30 mg / m 2 BSA. Calvert's formula should be used to calculate the correct clinical dose.

[0143] In one embodiment, the combination therapy comprises administering panobinostat and dasatinib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of dasatinib or a pharmaceutically acceptable salt thereof.

[0144] Dasatinib or a pharmaceutically acceptable salt thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0145] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with dasatinib or a pharmaceutically acceptable salt thereof for treating cholangiocarcinoma in the subject.

[0146] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject in combination with dasatinib or a pharmaceutically acceptable salt thereof to treat cholangiocarcinoma in the subject.

[0147] In some embodiments, the combination product of panobinostat and dasatinib is a combined preparation, such as a pharmaceutical composition comprising panobinostat and dasatinib in a single dosage form (eg, tablet or capsule).

[0148] In some embodiments, the combination therapy of panobinostat and dasatinib is used to treat extrahepatic bile duct cancer.

[0149] In some embodiments, the combination therapy of panobinostat and dasatinib is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0150] Dasatinib (N-(2-chloro-6-methylphenyl)-2-({6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl}amino)-1,3-thiazole-5-carboxamide) is a protein kinase inhibitor disclosed in WO2000 / 062778 (Formula I). ​​Dasatinib has the structure shown below. Dasatinib can be obtained from Bristol-Myers Squibb. The term "dasatinib" includes pharmaceutically acceptable salts and hydrates thereof.

[0151]

[0152] Pharmaceutical compositions of dasatinib are well known in the art, for example as described in WO2000 / 062778, WO2007 / 035874 and WO2015 / 181573 (all three of which are incorporated herein by reference) and any such compositions may be used in the methods, compositions and uses of the present invention.

[0153] In a preferred embodiment, the pharmaceutical composition comprising dasatinib is formulated for oral administration.

[0154] A preferred embodiment of using panobinostat in combination with dasatinib to treat cholangiocarcinoma is oral administration of panobinostat and dasatinib.

[0155] Thus, in some embodiments, panobinostat and dasatinib can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and dasatinib can be administered as a combined pharmaceutical formulation in one dosage form (e.g., tablet or capsule).

[0156] A pharmaceutical formulation comprising panobinostat and dasatinib in the same combined formulation (eg tablet or capsule) for use in the treatment of cholangiocarcinoma (a pharmaceutical composition as defined herein) is an aspect of the invention.

[0157] The most preferred aspect of the combined administration of panobinostat and dasatinib for the treatment of cholangiocarcinoma is the administration of dasatinib in the form of an oral formulation comprising dasatinib monohydrate.

[0158] According to the present invention, a typical oral formulation of dasatinib for treating cholangiocarcinoma comprises at least one of the following excipients: lactose, mannitol, microcrystalline cellulose, hydroxypropyl methylcellulose (HPMC), croscarmellose sodium, magnesium stearate, sodium lauryl sulfate, polyethylene glycol and silicon dioxide.

[0159] In some embodiments, the clinical dose of panobinostat combined with dasatinib for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0160] In some embodiments, the clinical dose of dasatinib combined with panobinostat for the treatment of cholangiocarcinoma should generally be 10 mg / day to 200 mg / day.

[0161] In one embodiment, the combination therapy comprises administering panobinostat and doxorubicin. Accordingly, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of doxorubicin or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0162] Doxorubicin or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0163] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with doxorubicin or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcin in the subject.

[0164] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcin in combination with doxorubicin or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0165] The combination therapy of panobinostat and doxorubicin can be used to treat intrahepatic cholangiocarcinoma or extrahepatic cholangiocarcinoma. In some embodiments, the combination therapy of panobinostat and doxorubicin is used to treat intrahepatic cholangiocarcinoma.

[0166] In some embodiments, the combination therapy of panobinostat and doxorubicin is used to treat a subject having a cholangiocarcin tumor having one or more characteristics specific to the CC-SW-1 cell line, the HuCC-T1 cell line, the EFI-1 cell line, and / or the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0167] Doxorubicin ((1S,3S)-3-hydroxyacetyl-3,5,12-trihydroxy-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetracen-1-yl-3-amino-2,3,6-trideoxy-α-L-lyxose-hexapyranoside) is a cytotoxic antibiotic drug substance having the structure shown below. Doxorubicin is widely available, for example, from Janssen and Pfizer.

[0168] The term "doxorubicin" includes pharmaceutically acceptable salts, solvates and hydrates thereof.

[0169]

[0170] Liquid pharmaceutical compositions of doxorubicin are well known in the art and any such compositions may be used in the methods, compositions and uses of the invention.

[0171] In some embodiments, the composition comprising doxorubicin is a "ready-to-use" formulation that comprises doxorubicin in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0172] In a preferred embodiment, the pharmaceutical composition comprising doxorubicin is formulated for parenteral administration.

[0173] A preferred embodiment of the treatment of cholangiocarcin using panobinostat in combination with doxorubicin is to administer panobinostat orally and doxorubicin as an injection or infusion.

[0174] In some embodiments, the clinical dose of panobinostat combined with doxorubicin for the treatment of cholangiocarcin is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0175] In some embodiments, the clinical dose of doxorubicin combined with panobinostat for the treatment of cholangiocarcin is generally the same as the dose range currently used for doxorubicin for other indications, such as 10 mg / m2 every 2-4 weeks. 2 Body surface area (BSA) up to 100 mg / m 2 Body surface area, preferably 40 mg / m 2 BSA to 75 mg / m 2 BSA.

[0176] In one embodiment, the combination therapy comprises administering panobinostat and gemcitabine. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of gemcitabine or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0177] Gemcitabine or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0178] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with gemcitabine or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0179] In another embodiment, the present invention provides use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with gemcitabine or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0180] The combination therapy of panobinostat and gemcitabine can be used to treat intrahepatic cholangiocarcinoma or extrahepatic cholangiocarcinoma. In some embodiments, the combination therapy of panobinostat and gemcitabine is used to treat intrahepatic cholangiocarcinoma.

[0181] In some embodiments, the combination therapy of panobinostat and gemcitabine is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, the HuCC-T1 cell line, the EGI-1 cell line, and / or the TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or the HuCC-T1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0182] Gemcitabine (4-amino-1-(2-deoxy-2,2-difluoro-β-D-erythro-pentofuranosyl)pyrimidin-2(1H)-one) is a nucleoside analog having the structure shown below. Gemcitabine is widely available, for example from Eli Lilly and Company. or Sigma-Aldrich, St. Louis, Missouri, USA. The term "gemcitabine" includes pharmaceutically acceptable salts, solvates and hydrates thereof. The pharmaceutically acceptable salt is preferably as defined above, preferably the hydrochloride.

[0183]

[0184] Liquid pharmaceutical compositions of gemcitabine are well known in the art and any such compositions may be used in the methods, compositions and uses of the present invention.

[0185] In some embodiments, the composition comprising gemcitabine is a "ready-to-use" formulation that comprises gemcitabine in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0186] In a preferred embodiment, the pharmaceutical composition comprising gemcitabine is formulated for parenteral administration.

[0187] A preferred embodiment of the treatment of cholangiocarcinoma using panobinostat in combination with gemcitabine is to administer panobinostat orally and gemcitabine as an injection or infusion.

[0188] In some embodiments, the clinical dose of panobinostat combined with gemcitabine for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0189] In some embodiments, the clinical dose of gemcitabine combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for gemcitabine for other indications, such as 500 mg / m 2 Up to 1500 mg / m 2 (mg of gemcitabine / m 2 Conveniently, 900 mg / m 2 Up to 1100 mg / m 2 Conveniently, gemcitabine may be administered in less than 1 hour, such as 15 minutes to 45 minutes, such as about 30 minutes, or over a longer period of time, such as 1 hour to 12 hours.

[0190] In one embodiment, the combination therapy comprises administering panobinostat and methotrexate. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of methotrexate or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0191] Methotrexate or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0192] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with methotrexate or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0193] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with methotrexate or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0194] In some embodiments, the combination product of panobinostat and methotrexate is a combined preparation, such as a pharmaceutical composition comprising panobinostat and methotrexate in a single dosage form (eg, tablet or capsule).

[0195] In some embodiments, combination therapy of panobinostat and methotrexate is used to treat extrahepatic bile duct cancer.

[0196] In some embodiments, combination therapy of panobinostat and methotrexate is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, HuCC-T1 cell line, and / or TFK-1 cell line, preferably specific to the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0197] Methotrexate (N-[4-[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamic acid) is a folic acid derivative (antimetabolite) having the structure shown below. Methotrexate is widely available, for example from Hospira. The term "methotrexate" includes pharmaceutically acceptable salts, solvates and hydrates thereof. The pharmaceutically acceptable salt is preferably as defined above, preferably the sodium salt.

[0198]

[0199] Liquid and solid pharmaceutical compositions of methotrexate are well known in the art and any such compositions may be used in the methods, compositions and uses of the present invention.

[0200] In some embodiments, the composition comprising methotrexate is a "ready-to-use" formulation, which contains methotrexate in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0201] Thus, in some embodiments, the pharmaceutical composition comprising methotrexate is formulated for parenteral administration, such as injection or infusion. In these embodiments, the methotrexate may be provided in the form of a salt, preferably in the form of a sodium salt.

[0202] However, in some embodiments, the pharmaceutical composition comprising methotrexate is formulated for oral administration, such as a tablet or capsule.

[0203] In some embodiments, the use of panobinostat in combination with methotrexate to treat cholangiocarcinoma is oral administration of panobinostat and administration of methotrexate as an injection or infusion.

[0204] In other embodiments, the treatment of cholangiocarcinoma with the combination of panobinostat and methotrexate is oral administration of panobinostat and methotrexate.

[0205] Thus, in some embodiments, panobinostat and methotrexate can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and methotrexate can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0206] Therefore, a pharmaceutical formulation (pharmaceutical composition) comprising panobinostat and methotrexate in the same combined formulation (eg tablet or capsule) for use in the treatment of cholangiocarcinoma is a further aspect of the present invention.

[0207] In some embodiments, the clinical dose of panobinostat combined with methotrexate for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0208] In some embodiments, the clinical dose of methotrexate combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for methotrexate for other indications. For example, in some embodiments, the dose range of methotrexate can be 2.5 mg / m2 administered weekly. 2 BSA to 50 mg / m 2 BSA, for example 7.5 mg / m 2 BSA to 25 mg / m 2 BSA.

[0209] In one embodiment, the combination therapy comprises administering panobinostat and topotecan. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of topotecan or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0210] Topotecan or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0211] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with topotecan or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0212] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with topotecan or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0213] In some embodiments, the combination product of panobinostat and topotecan is a combined preparation, such as a pharmaceutical composition comprising panobinostat and topotecan in a single dosage form (eg, tablet or capsule).

[0214] The combination of panobinostat and topotecan can be used to treat intrahepatic or extrahepatic cholangiocarcinoma.

[0215] In some embodiments, the combination therapy of panobinostat and topotecan is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0216] Topotecan (9-[(dimethylamino)methyl]-10-hydroxy-(4S)-camptothecin) is a topoisomerase inhibitor having the structure shown below. Topotecan is widely available, for example, from Actavis. The term "topotecan" includes pharmaceutically acceptable salts, solvates and hydrates thereof. The pharmaceutically acceptable salt is preferably as defined above, preferably the hydrochloride.

[0217]

[0218] Liquid and solid pharmaceutical compositions of topotecan are well known in the art and any such compositions may be used in the methods, compositions and uses of the present invention.

[0219] In some embodiments, the composition comprising topotecan is a "ready-to-use" formulation, which contains topotecan in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0220] Thus, in some embodiments, the pharmaceutical composition comprising topotecan is formulated for parenteral administration, such as injection or infusion.

[0221] However, in some embodiments, the pharmaceutical composition comprising topotecan is formulated for oral administration, such as a tablet or capsule.

[0222] In some embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with topotecan is by orally administering panobinostat and administering topotecan as an injection or infusion.

[0223] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with topotecan is oral administration of panobinostat and topotecan.

[0224] Thus, in some embodiments, panobinostat and topotecan can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and topotecan can be administered as a combined pharmaceutical formulation (ie, pharmaceutical composition) in one dosage form (e.g., tablet or capsule).

[0225] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and topotecan in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0226] In some embodiments, the clinical dose of panobinostat combined with topotecan for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0227] In some embodiments, the clinical dose of topotecan combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for topotecan for other indications. For example, in some embodiments, the dose range of topotecan can be 0.25 mg / m 2 BSA to 3 mg / m 2 BSA, for example 0.75 mg / m 2 BSA to 1.50 mg / m 2 BSA, as defined above, is administered daily or at least twice a week.

[0228] In one embodiment, the combination therapy comprises administering panobinostat and trametinib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of trametinib or a pharmaceutically acceptable salt thereof.

[0229] Trametinib or a pharmaceutically acceptable salt thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0230] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with trametinib or a pharmaceutically acceptable salt thereof for treating cholangiocarcinoma in the subject.

[0231] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for separate, simultaneous or sequential use or administration to a subject in combination with trametinib or a pharmaceutically acceptable salt thereof to treat cholangiocarcinoma in the subject.

[0232] In some embodiments, the combination product of panobinostat and trametinib is a combined preparation, such as a pharmaceutical composition comprising panobinostat and trametinib in a single dosage form (eg, tablet or capsule).

[0233] The combination of panobinostat and trametinib can be used to treat extrahepatic cholangiocarcinoma or intrahepatic cholangiocarcinoma.

[0234] In some embodiments, the combination therapy of panobinostat and trametinib is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0235] Trametinib is a tyrosine kinase inhibitor with affinity for mitogen-activated protein kinases and having a structure as shown below. Trametinib is available from Novartis. The term "trametinib" includes pharmaceutically acceptable salts thereof as defined elsewhere herein. In some embodiments, trametinib is provided in the form of trametinib dimethyl sulfoxide.

[0236]

[0237] Liquid pharmaceutical compositions of trametinib are well known in the art and any such compositions may be used in the methods, compositions and uses of the invention.

[0238] In a preferred embodiment, the pharmaceutical composition comprising trametinib is formulated for oral administration (eg, tablets or capsules).

[0239] A preferred embodiment of the treatment of cholangiocarcinoma using panobinostat in combination with trametinib is oral administration of panobinostat and trametinib.

[0240] Therefore, in some embodiments, panobinostat and trametinib can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and trametinib can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0241] A pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and trametinib in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0242] In a preferred aspect of the invention, the combined administration of panobinostat and trametinib for the treatment of cholangiocarcinoma involves the use of an oral formulation of trametinib, wherein trametinib is optionally in the form of a dimethyl sulfate solvate, and the oral formulation comprises one or more of the following excipients: mannitol, microcrystalline cellulose, hydroxypropyl methylcellulose (HPMC), croscarmellose sodium, magnesium stearate, sodium lauryl sulfate, and silicon dioxide.

[0243] In some embodiments, the clinical dose of panobinostat combined with trametinib for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0244] In some embodiments, the clinical dose of trametinib combined with panobinostat for the treatment of cholangiocarcinoma is generally 0.1 mg to 10 mg per day, more preferably 0.5 mg to 5 mg, or at least twice a week, for example 2-6 times, 2-5 times or 2-4 times. In a preferred embodiment, the clinical dose is a single dose formulation, such as a tablet or capsule.

[0245] In one embodiment, the combination therapy comprises administering panobinostat and combretastat A4. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of combretastat A4 or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0246] Combretastatin A4 or a pharmaceutically acceptable salt, solvate or hydrate thereof can be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0247] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with combretastatin A4 or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0248] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with combretastatin A4 or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0249] In some embodiments, the combination product of panobinostat and combretastat A4 is a combined preparation, such as a pharmaceutical composition comprising panobinostat and combretastat A4 in a single dosage form (eg, tablet or capsule).

[0250] In some embodiments, combination therapy of panobinostat and combretastatin A4 is used to treat intrahepatic cholangiocarcinoma.

[0251] In some embodiments, combination therapy of panobinostat and combretastatin A4 is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0252] Combretastatin A4 (2-methoxy-5-[(Z)-2-(3,4,5-trimethoxy-phenyl)-vinyl]-phenol) is a stilbene compound having the structure shown below. It can be isolated from the shrub (Combretum caffrum). The term "combretastatin A4" includes its pharmaceutically acceptable salts, solvates and hydrates. Pharmaceutically acceptable salts, solvates and hydrates are preferably as defined above. In some embodiments, combretastatin A4 is provided in the form of a water-soluble ester, for example, in the form of a water-soluble phosphate ester.

[0253]

[0254] Combretastatin A4 can be provided in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0255] In some embodiments, the composition comprising combretastatin A4 is a "ready-to-use" formulation, which contains combretastatin A4 in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0256] Thus, in some embodiments, the pharmaceutical composition comprising combretastatin A4 is formulated for parenteral administration, such as injection or infusion.

[0257] However, in some embodiments, the pharmaceutical composition comprising combretastatin A4 is formulated for oral administration, such as a tablet or capsule.

[0258] In some embodiments, the combination of panobinostat and combretastat A4 for treating cholangiocarcinoma is oral administration of panobinostat and administration of combretastat A4 as an injection or infusion.

[0259] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with combretastat A4 is oral administration of panobinostat and combretastat A4.

[0260] Thus, in some embodiments, panobinostat and combretastatin A4 can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and combretastatin A4 can be administered as a combined pharmaceutical formulation (i.e., pharmaceutical composition) in one dosage form (e.g., tablet or capsule).

[0261] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and combretastatin A4 in the same combined formulation (eg, tablet or capsule) for use in treating cholangiocarcinoma is a further aspect of the present invention.

[0262] In some embodiments, the clinical dose of panobinostat combined with combretastatin A4 for treating cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0263] In some embodiments, the clinical dose of combretastatin A4 combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for combretastatin A4 for other indications. For example, in some embodiments, the dose range of combretastatin A4 can be 5 mg / m 2 BSA to 100 mg / m 2 BSA, for example 20 mg / m 2 BSA to 85 mg / m 2 BSA, as defined above, is administered daily or at least twice a week.

[0264] In one embodiment, the combination therapy comprises administering panobinostat and SB-743921. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of SB-743921 or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0265] SB-743921 or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0266] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with SB-743921 or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0267] In another embodiment, the present invention provides use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with SB-743921 or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0268] The combination therapy of panobinostat and SB-743921 can be used to treat intrahepatic cholangiocarcinoma or extrahepatic cholangiocarcinoma. In some embodiments, the combination therapy of panobinostat and SB-743921 is used to treat extrahepatic cholangiocarcinoma.

[0269] In some embodiments, the combination therapy of panobinostat and SB-743921 is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, the HuCC-T1 cell line, the EGI-1 cell line, and / or the TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or the EGI-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0270] SB-743921 is an inhibitor of the mitotic kinesin KSP having the structure shown below. The term "SB-743921" includes pharmaceutically acceptable salts, solvates and hydrates thereof. The pharmaceutically acceptable salt is preferably as defined above, preferably a hydrochloride.

[0271]

[0272] Liquid pharmaceutical compositions of SB-743921 are well known in the art and any such compositions may be used in the methods, compositions and uses of the present invention.

[0273] In some embodiments, the composition comprising SB-743921 is a "ready-to-use" formulation that contains SB-743921 in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0274] In a preferred embodiment, the pharmaceutical composition comprising SB-743921 is formulated for parenteral administration.

[0275] A preferred embodiment of the treatment of cholangiocarcinoma using panobinostat in combination with SB-743921 is to administer panobinostat orally and SB-743921 as an injection or infusion.

[0276] In some embodiments, the clinical dose of panobinostat combined with SB-743921 for treating cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0277] In some embodiments, the clinical dose of SB-743921 in combination with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for SB-743921 for other indications. For example, in some embodiments, the dose range of SB-743921 can be 1 mg / m2 administered weekly or monthly (e.g., every 1-4 weeks). 2 BSA to 10 mg / m 2 BSA.

[0278] In one embodiment, the combination therapy comprises administering panobinostat and dapolinet. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of dapolinet or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0279] Dapolinet or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0280] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with dapoline or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0281] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with dapolinet or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0282] In some embodiments, the combination product of panobinostat and dapolinet is a combined preparation, such as a pharmaceutical composition comprising panobinostat and dapolinet in a single dosage form (eg, tablet or capsule).

[0283] The combination therapy of panobinostat and dapolinet can be used to treat intrahepatic cholangiocarcinoma or extrahepatic cholangiocarcinoma. In some embodiments, the combination therapy of panobinostat and dapolinet is used to treat intrahepatic cholangiocarcinoma.

[0284] In some embodiments, the combination therapy of panobinostat and dapoline is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, the EGI-1 cell line, and / or the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0285] Dapoline ((E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-pyridin-3-ylprop-2-enamide) inhibits nicotinamide phosphoribosyltransferase (NMPRTase) and has the structure shown below. The term "dapoline" includes pharmaceutically acceptable salts, solvates and hydrates thereof. The pharmaceutically acceptable salt is preferably as defined above, preferably the hydrochloride.

[0286]

[0287] Dapolinet may be provided for use in the methods, compositions and uses of the present invention in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition.

[0288] In some embodiments, the composition comprising dapolinel is a "ready-to-use" formulation that contains dapolinel in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0289] Thus, in some embodiments, the pharmaceutical composition comprising dapolinel is formulated for parenteral administration, such as injection or infusion.

[0290] However, in some embodiments, the pharmaceutical composition comprising dapolinel is formulated for oral administration, such as a tablet or capsule.

[0291] In some embodiments, the combination of panobinostat and dapolinet for treating cholangiocarcinoma is oral administration of panobinostat and administration of dapolinet as an injection or infusion.

[0292] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with dapolinet is oral administration of panobinostat and dapolinet.

[0293] Thus, in some embodiments, panobinostat and dapolinet can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and dapolinet can be administered as a combined pharmaceutical formulation (i.e., pharmaceutical composition) in one dosage form (e.g., tablet or capsule).

[0294] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and dapolinet in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0295] In some embodiments, the clinical dose of panobinostat combined with dapoline for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0296] In some embodiments, the clinical dose of dapoline combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for dapoline for other indications. For example, in some embodiments, the dose range of dapoline can be 0.1 mg / m2 administered weekly or monthly. 2 BSA to 10 mg / m 2 BSA, for example, every 1-6 weeks, 1-5 weeks, 1-4 weeks, or 1-3 weeks.

[0297] In one embodiment, the combination therapy comprises administering panobinostat and ispins. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of ispins or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0298] Ispins or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0299] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with ispins or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0300] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with ispins or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0301] In some embodiments, the combination product of panobinostat and ispinas is a combined preparation, such as a pharmaceutical composition comprising panobinostat and ispinas in a single dosage form (eg, tablet or capsule).

[0302] In some embodiments, the combination therapy of panobinostat and ispinast is used to treat extrahepatic bile duct cancer.

[0303] In some embodiments, the combination therapy of panobinostat and ispinas is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, the EGI-1 cell line, and / or the TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0304] Ispins (N-(3-aminopropyl)-N-[(1R)-1-[7-chloro-4-oxo-3-(benzyl)-2-quinazolinyl]-2-methylpropyl]-4-methylbenzamide) is derived from quinazolinone and selectively inhibits mitotic motor proteins and spindle kinesin (KSP). Ispins has the structure shown below. The term "ispins" includes pharmaceutically acceptable salts, solvates and hydrates thereof. For example, in some embodiments, ispins can be in the form of a hydrochloride salt. In some preferred embodiments, ispins is in the form of a free compound.

[0305]

[0306] Ispins may be provided for use in the methods, compositions and uses of the present invention in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition.

[0307] In some embodiments, the composition comprising Ispins is a "ready-to-use" formulation that contains Ispins in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0308] Thus, in some embodiments, pharmaceutical compositions comprising Ispins are formulated for parenteral administration, such as injection or infusion.

[0309] However, in some embodiments, pharmaceutical compositions comprising Ispins are formulated for oral administration, such as tablets or capsules.

[0310] In some embodiments, the use of panobinostat in combination with ispins to treat cholangiocarcinoma is oral administration of panobinostat and administration of ispins as an injection or infusion.

[0311] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with ispinas is oral administration of panobinostat and ispinas.

[0312] Thus, in some embodiments, panobinostat and ispins can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and ispins can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0313] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and ispinast in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0314] In some embodiments, the clinical dose of panobinostat combined with ispins for treating cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0315] In some embodiments, the clinical dose of Ispins combined with Panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for Ispins for other indications. For example, in some embodiments, the dose range of Ispins can be 5 mg / m2 administered weekly or monthly. 2 BSA to 30 mg / m 2 BSA, for example, every 1-6 weeks, 1-5 weeks, 1-4 weeks, or 1-3 weeks.

[0316] In one embodiment, the combination therapy comprises administering panobinostat and lumispivot. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of lumispivot or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0317] Lumispir or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0318] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with lumirastat or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0319] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with lumirastat or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0320] In some embodiments, the combination product of panobinostat and lumibenostat is a combined preparation, such as a pharmaceutical composition comprising panobinostat and lumibenostat in a single dosage form (eg, tablet or capsule).

[0321] In some embodiments, the combination therapy of panobinostat and lumispivir is used to treat extrahepatic cholangiocarcinoma.

[0322] In some embodiments, the combination therapy of panobinostat and lumiraspir is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the EGI-1 cell line and / or TFK-1 cell line (preferably specific to the EGI-1 cell line), such as one or more genetic markers, growth rate, and / or cell morphology.

[0323] Lumispir (5-(2,4-dihydroxy-5-isopropyl-phenyl)-N-ethyl-4-[4-(morpholinomethyl)phenyl]isoxazole-3-carboxamide) is an HSP90 inhibitor having a structure as shown below. The term "Lumispir" includes pharmaceutically acceptable salts, solvates and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above. For example, in some embodiments, Lumispir may be in the form of a hydrochloride or a mesylate.

[0324]

[0325] Lumispir can be provided in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0326] In some embodiments, the composition comprising lumispiv is a "ready-to-use" formulation that contains lumispiv in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0327] Thus, in some embodiments, the pharmaceutical composition comprising lumispivir is formulated for parenteral administration, such as injection or infusion.

[0328] However, in some embodiments, the pharmaceutical composition comprising lumispivir is formulated for oral administration, such as a tablet or capsule.

[0329] In some embodiments, the use of panobinostat in combination with lumibigostat to treat cholangiocarcinoma is oral administration of panobinostat and administration of lumibigostat as an injection or infusion.

[0330] In other embodiments, the use of panobinostat in combination with lumibinostat to treat cholangiocarcinoma is oral administration of panobinostat and lumibinostat.

[0331] Thus, in some embodiments, panobinostat and lumispivot can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and lumispivot can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0332] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and lumispivir in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0333] In some embodiments, the clinical dose of panobinostat combined with lumispivir for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0334] In some embodiments, the clinical dose of lumispi combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for lumispi for other indications. For example, in some embodiments, the dose range of lumispi can be 5 mg / m2 administered weekly. 2 BSA to 150 mg / m 2 BSA, for example 40 mg / m 2 BSA to 70 mg / m 2 BSA, for example, every 1-4 weeks, 1-3 weeks, or 1-2 weeks.

[0335] In one embodiment, the combination therapy comprises administering panobinostat and molibis. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of molibis or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0336] Molibis or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0337] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with molibox or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0338] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for use or administration to a subject separately, simultaneously or sequentially in combination with moliboxetine or a pharmaceutically acceptable salt, solvate or hydrate thereof to treat cholangiocarcinoma in the subject.

[0339] In some embodiments, the combination product of panobinostat and molibox is a combined preparation, such as a pharmaceutical composition comprising panobinostat and molibox in a single dosage form (eg, tablet or capsule).

[0340] In some embodiments, the combination therapy of panobinostat and molibex is used to treat extrahepatic cholangiocarcinoma.

[0341] In some embodiments, the combination therapy of panobinostat and molibis is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the EGI-1 cell line and / or the TFK-1 cell line (preferably specific to the TFK-1 cell line), such as one or more genetic markers, growth rate, and / or cell morphology.

[0342] Molib (2-[(4S)-6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]-N-ethylacetamide) is an inhibitor of the BET (Bromodomain and Extra-Terminal) family of bromodomain-containing proteins having the structure shown below. The term "molib" includes pharmaceutically acceptable salts, solvates and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above. For example, in some embodiments, molib can be in the form of a hydrochloride or a methanesulfonate.

[0343]

[0344] Molybus can be provided for use in the methods, compositions and uses of the present invention in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition.

[0345] In some embodiments, the composition comprising molybus is a "ready-to-use" formulation that contains molybus in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0346] Thus, in some embodiments, pharmaceutical compositions comprising molybus are formulated for parenteral administration, such as injection or infusion.

[0347] However, in some embodiments, the pharmaceutical composition comprising molybus is formulated for oral administration, such as a tablet or capsule.

[0348] In some embodiments, the use of panobinostat in combination with molibex to treat cholangiocarcinoma is oral administration of panobinostat and administration of molibex as an injection or infusion.

[0349] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with molibex is oral administration of panobinostat and molibex.

[0350] Thus, in some embodiments, panobinostat and molibox can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and molibox can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0351] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and molibis in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0352] In some embodiments, the clinical dose of panobinostat combined with molibex for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0353] In some embodiments, the clinical dose of molibux in combination with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for molibux for other indications. For example, in some embodiments, the dose range of molibux can be 5 mg to 150 mg, such as 10 mg to 80 mg, administered daily.

[0354] In one embodiment, the combination therapy comprises administering panobinostat and pelitinib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of pelitinib or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0355] Pelitatinib or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0356] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with peritinib or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0357] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for use in combination with peritinib or a pharmaceutically acceptable salt, solvate or hydrate thereof for separate, simultaneous or sequential use or administration to a subject to treat cholangiocarcinoma in the subject.

[0358] In some embodiments, the combination product of panobinostat and peritinib is a combined preparation, such as a pharmaceutical composition comprising panobinostat and peritinib in a single dosage form (eg, tablet or capsule).

[0359] In some embodiments, the combination therapy of panobinostat and peritinib is used to treat extrahepatic cholangiocarcinoma.

[0360] In some embodiments, the combination therapy of panobinostat and trametinib is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0361] Pelitanib ((2E)-N-(4-((3-chloro-4-fluorophenyl)amino)-3-cyano-7-ethoxy-6-quinolyl)-4-(dimethylamino)-2-butenamide) is an irreversible inhibitor of epidermal growth factor receptor (EGFR) having the structure shown below. The term "pelitanib" includes pharmaceutically acceptable salts, solvates and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above. For example, in some embodiments, pelitanib can be in the form of an acid salt, such as a hydrochloride or a mesylate.

[0362]

[0363] Peritinib can be provided for use in the methods, compositions and uses of the present invention in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition.

[0364] In some embodiments, the composition comprising peritinib is a "ready-to-use" formulation, which contains peritinib in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0365] Thus, in some embodiments, pharmaceutical compositions comprising peritinib are formulated for parenteral administration, such as injection or infusion.

[0366] However, in some embodiments, the pharmaceutical composition comprising peritinib is formulated for oral administration, such as a tablet or capsule.

[0367] In some embodiments, the use of panobinostat in combination with peritinib to treat cholangiocarcinoma is oral administration of panobinostat and administration of peritinib as an injection or infusion.

[0368] In other embodiments, the use of panobinostat in combination with peritinib to treat cholangiocarcinoma is oral administration of panobinostat and peritinib.

[0369] Thus, in some embodiments, panobinostat and peritinib can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and peritinib can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0370] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and peritinib in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0371] In some embodiments, the clinical dose of panobinostat combined with peritinib for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0372] In some embodiments, the clinical dose of peritinib combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for peritinib for other indications. For example, in some embodiments, the dose range of peritinib can be 10 mg to 100 mg, such as 25 mg to 75 mg, administered daily.

[0373] In one embodiment, the combination therapy comprises administering panobinostat and triptolide. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of triptolide or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0374] Triptolide or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0375] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with triptolide or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0376] In another embodiment, the present invention provides use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for use or administration to a subject, either separately, simultaneously or sequentially, in combination with triptolide or a pharmaceutically acceptable salt, solvate or hydrate thereof, to treat cholangiocarcinoma in the subject.

[0377] In some embodiments, the combination product of panobinostat and triptolide is a combined preparation, such as a pharmaceutical composition comprising panobinostat and triptolide in a single dosage form (eg, tablet or capsule).

[0378] In some embodiments, combination therapy of panobinostat and triptolide is used to treat extrahepatic bile duct cancer.

[0379] In some embodiments, combination therapy of panobinostat and triptolide is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0380] Triptolide is a diterpene epoxide having the structure shown below. The term "triptolide" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. In some embodiments, triptolide may be provided in the form of a water-soluble prodrug.

[0381]

[0382] Triptolide may be provided in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0383] In some embodiments, the composition comprising triptolide is a "ready-to-use" formulation that contains triptolide in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0384] Thus, in some embodiments, pharmaceutical compositions comprising triptolide are formulated for parenteral administration, such as injection or infusion.

[0385] However, in some embodiments, the pharmaceutical composition comprising triptolide is formulated for oral administration, such as a tablet or capsule.

[0386] In some embodiments, the use of panobinostat in combination with triptolide to treat cholangiocarcinoma is oral administration of panobinostat and administration of triptolide as an injection or infusion.

[0387] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with triptolide is oral administration of panobinostat and triptolide.

[0388] Thus, in some embodiments, panobinostat and triptolide can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and triptolide can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0389] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and triptolide in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0390] In some embodiments, the clinical dose of panobinostat combined with triptolide for treating cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0391] In some embodiments, the clinical dose of triptolide combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for triptolide for other indications. For example, in some embodiments, the dose range of triptolide can be 10 mg to 200 mg, such as 25 mg to 150 mg, administered daily.

[0392] In one embodiment, the combination therapy comprises administering panobinostat and BI 2536. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of BI 2536 or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0393] BI 2536 or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0394] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with BI 2536 or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0395] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with BI2536 or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0396] In some embodiments, the combination product of panobinostat and BI 2536 is a combined preparation, such as a pharmaceutical composition comprising panobinostat and BI 2536 in a single dosage form (eg, tablet or capsule).

[0397] In some embodiments, the combination therapy of panobinostat and BI 2536 is used to treat extrahepatic cholangiocarcinoma.

[0398] In some embodiments, combination therapy of panobinostat and BI 2536 is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0399] BI 2536 is an inhibitor of PLK1 (polo-like kinase 1) protein having the structure shown below. The term "BI2536" includes pharmaceutically acceptable salts, solvates and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above. For example, in some embodiments, BI 2536 can be in the form of an acid salt, such as a hydrochloride.

[0400]

[0401] BI 2536 can be provided for the methods, compositions and uses of the present invention in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition.

[0402] In some embodiments, the composition comprising BI 2536 is a "ready-to-use" formulation that contains BI 2536 in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0403] Thus, in some embodiments, pharmaceutical compositions comprising BI 2536 are formulated for parenteral administration, such as injection or infusion.

[0404] However, in some embodiments, the pharmaceutical composition comprising BI 2536 is formulated for oral administration, such as a tablet or capsule.

[0405] In some embodiments, the combination of panobinostat and BI 2536 for treating cholangiocarcinoma is oral administration of panobinostat and administration of BI 2536 as an injection or infusion.

[0406] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with BI 2536 is oral administration of panobinostat and BI 2536.

[0407] Thus, in some embodiments, panobinostat and BI 2536 can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and BI 2536 can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0408] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and BI2536 in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0409] In some embodiments, the clinical dose of panobinostat in combination with BI 2536 for treating cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0410] In some embodiments, the clinical dose of BI 2536 in combination with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for other indications of BI 2536. For example, in some embodiments, the dose range of BI 2536 can be 1 mg to 200 mg, such as 25 mg to 150 mg, administered daily.

[0411] In one embodiment, the combination therapy comprises administering panobinostat and datolicoxib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of datolicoxib or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0412] Datolicoxib or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0413] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with datolicoxib or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0414] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the preparation of a product for use or administration to a subject, either separately, simultaneously or sequentially, in combination with datolicoxib or a pharmaceutically acceptable salt, solvate or hydrate thereof, to treat cholangiocarcinoma in the subject.

[0415] In some embodiments, the combination product of panobinostat and datolicoxib is a combined preparation, such as a pharmaceutical composition comprising panobinostat and datolicoxib in a single dosage form (eg, tablet or capsule).

[0416] The combination therapy of panobinostat and datolicoxib can be used to treat extrahepatic cholangiocarcinoma or intrahepatic cholangiocarcinoma. In some embodiments, the combination therapy of panobinostat and datolicoxib is used to treat extrahepatic cholangiocarcinoma.

[0417] In some embodiments, the combination therapy of panobinostat and datolicoxib is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably specific to the CC-SW-1 cell line and / or TFK-1 cell line, most preferably specific to the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0418] Datolicoxib is a phosphoinositide-3-kinase inhibitor (PI3K inhibitor) and also inhibits mTOR. Datolicoxib has the structure shown below. The term "datolicoxib" includes pharmaceutically acceptable salts, solvates and hydrates thereof.

[0419]

[0420] Datolicoxib

[0421] Datolicoxib may be provided for use in the methods, compositions and uses of the present invention in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition.

[0422] In some embodiments, the composition comprising datolicoxib is a "ready-to-use" formulation that contains datolicoxib in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0423] Thus, in some embodiments, pharmaceutical compositions comprising datolicoxib are formulated for parenteral administration, such as injection or infusion.

[0424] However, in some embodiments, the pharmaceutical composition comprising datolicoxib is formulated for oral administration, such as a tablet or capsule.

[0425] In some embodiments, the use of panobinostat in combination with datolicoxib to treat cholangiocarcinoma is oral administration of panobinostat and administration of datolicoxib as an injection or infusion.

[0426] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with datolicoxib is oral administration of panobinostat and datolicoxib.

[0427] Thus, in some embodiments, panobinostat and datolicoxib can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and datolicoxib can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0428] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and datolicoxib in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0429] In some embodiments, the clinical dose of panobinostat combined with datolicoxib for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0430] In some embodiments, the clinical dose of datolicoxib combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for datolicoxib for other indications. For example, in some embodiments, the dose range of datolicoxib can be 100 mg to 1200 mg, such as 200 mg to 800 mg, administered daily.

[0431] In one embodiment, the combination therapy comprises administering panobinostat and obaclava. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of obaclava or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0432] Obaclava or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0433] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with obaclava or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0434] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with obaclava or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0435] In some embodiments, the combination product of panobinostat and obaclava is a combined preparation, such as a pharmaceutical composition comprising panobinostat and obaclava in a single dosage form (eg, tablet or capsule).

[0436] In some embodiments, the combination therapy of panobinostat and obaclava is used to treat extrahepatic bile duct cancer.

[0437] In some embodiments, the combination therapy of panobinostat and obaclava is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the EGI-1 cell line and / or TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0438] Obakla is an inhibitor of Bcl-2 family proteins having the structure shown below. The term "obakla" includes pharmaceutically acceptable salts, solvates and hydrates thereof. The pharmaceutically acceptable salt is preferably obakla mesylate.

[0439]

[0440] Obakra can be provided for the methods, compositions and uses of the present invention in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition.

[0441] In some embodiments, the composition comprising obaclava is a "ready-to-use" formulation that contains obaclava in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0442] Thus, in some embodiments, pharmaceutical compositions comprising obaclava are formulated for parenteral administration, such as injection or infusion.

[0443] However, in some embodiments, the pharmaceutical composition comprising obaclava is formulated for oral administration, such as a tablet or capsule.

[0444] In some embodiments, the combination of panobinostat and obaclava for treating cholangiocarcinoma is oral administration of panobinostat and administration of obaclava as an injection or infusion.

[0445] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with obaclava is oral administration of panobinostat and obaclava.

[0446] Thus, in some embodiments, panobinostat and obaclava can be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and obaclava can be administered as a combined pharmaceutical formulation (i.e., a pharmaceutical composition) in one dosage form (e.g., a tablet or capsule).

[0447] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and obaclava in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0448] In some embodiments, the clinical dose of panobinostat combined with obaclava for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0449] In some embodiments, the clinical dose of obaclave combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for obaclave for other indications. For example, in some embodiments, the dose range of obaclave can be 5 mg / m2 / day. 2 BSA to 50 mg / m 2 BSA, e.g. 10 mg / m 2 BSA to 20 mg / m 2 BSA.

[0450] In one embodiment, the combination therapy comprises administering panobinostat and ilisimostat. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt, solvate or hydrate thereof and a therapeutically effective amount of ilisimostat or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0451] Elisemostat or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0452] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt, solvate or hydrate thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with ilisimostat or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0453] In another embodiment, the present invention provides use of panobinostat or a pharmaceutically acceptable salt, solvate or hydrate thereof in the preparation of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with ilisimostat or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0454] In some embodiments, the combination product of panobinostat and ilisimol is a combined preparation, such as a pharmaceutical composition comprising panobinostat and ilisimol in a single dosage form (eg, injection or infusion).

[0455] The combination of panobinostat and ilisimol can be used to treat extrahepatic cholangiocarcinoma or intrahepatic cholangiocarcinoma.

[0456] In some embodiments, the combination therapy of panobinostat and ilisimol is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line, the HuCC-T1 cell line, the EGI-1 cell line, and / or the TFK-1 cell line, preferably specific to the TFK-1 cell line, such as one or more genetic markers, growth rate, and / or cell morphology.

[0457] Elisimol (1-N', 3-N'-bis(phenylthiocarbonyl)-1-N', 3-N'-dimethylmalonohydrazide) induces oxidative stress, producing high levels of reactive oxygen species (ROS), such as hydrogen peroxide, in cancer cells and normal cells. Elisimol has the structure shown below. The term "elismol" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. In some embodiments, Elisimol is provided in the form of a sodium salt.

[0458]

[0459] Elisemomod may be provided in the form of a liquid pharmaceutical composition or a solid pharmaceutical composition for the methods, compositions and uses of the present invention. Elisemomod is described in WO2013071106, which is incorporated herein by reference.

[0460] In some embodiments, the composition comprising elisimomod is a "ready-to-use" formulation, which contains elisimomod in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0461] Thus, in some embodiments, the pharmaceutical composition comprising ilisimol is formulated for parenteral administration, such as injection or infusion.

[0462] However, in some embodiments, the pharmaceutical composition comprising ilisimol is formulated for oral administration, such as a tablet or capsule.

[0463] In some embodiments, the use of panobinostat in combination with elisimomod to treat cholangiocarcinoma is oral administration of panobinostat and administration of elisimomod as an injection or infusion.

[0464] In other embodiments, the treatment of cholangiocarcinoma with panobinostat in combination with elisimostat is oral administration of panobinostat and elisimostat.

[0465] Thus, in some embodiments, panobinostat and ilisimol can be administered in separate dosage forms (e.g., tablets or capsules). In some embodiments, panobinostat and ilisimol can be administered as a combined pharmaceutical formulation (i.e., pharmaceutical composition) in one dosage form (e.g., tablet or capsule).

[0466] Therefore, a pharmaceutical formulation (ie, a pharmaceutical composition) comprising panobinostat and ilisimol in the same combined formulation (eg, tablet or capsule) for treating cholangiocarcinoma is yet another aspect of the present invention.

[0467] In some embodiments, the clinical dose of panobinostat combined with ilisimol for treating cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0468] In some embodiments, the clinical dose of ilisimostat combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for ilisimostat for other indications. For example, in some embodiments, the dose range of ilisimostat can be 50 mg / m2 per day. 2 BSA to 300 mg / m 2 BSA, for example, 100 mg / m 2 BSA to 200 mg / m 2 BSA.

[0469] In one embodiment, the combination therapy comprises administering panobinostat and docetaxel. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of docetaxel or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0470] Docetaxel or a pharmaceutically acceptable salt, solvate or hydrate thereof may be administered separately, simultaneously or sequentially with a therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0471] Alternatively, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof as a product for separate, simultaneous or sequential use or administration to a subject in combination with docetaxel or a pharmaceutically acceptable salt, solvate or hydrate thereof for treating cholangiocarcinoma in the subject.

[0472] In another embodiment, the present invention provides use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a product for separate, simultaneous or sequential use or administration to a subject for treating cholangiocarcinoma in combination with docetaxel or a pharmaceutically acceptable salt, solvate or hydrate thereof.

[0473] In some embodiments, the combination therapy of panobinostat and docetaxel is used to treat intrahepatic cholangiocarcinoma.

[0474] In some embodiments, the combination therapy of panobinostat and docetaxel is used to treat a subject having a cholangiocarcinoma tumor having one or more characteristics specific to the CC-SW-1 cell line and / or TFK-1 cell line (preferably specific to the CC-SW-1 cell line), such as one or more genetic markers, growth rate, and / or cell morphology.

[0475] Docetaxel (N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetylpaclitaxel) is an antimitotic chemotherapeutic drug that reversibly binds tubulin with high affinity in a 1:1 stoichiometric ratio. Docetaxel has the structure listed below and is widely available, for example, from Actavis. The term "docetaxel" includes pharmaceutically acceptable solvates and hydrates thereof. In some embodiments, docetaxel is provided as docetaxel trihydrate.

[0476]

[0477] Liquid pharmaceutical compositions of docetaxel are well known in the art and any such compositions may be used in the methods, compositions and uses of the present invention.

[0478] In some embodiments, the composition comprising docetaxel is a "ready-to-use" formulation that contains docetaxel in dissolved or solubilized form and is intended for use as is or after further dilution in an intravenous diluent.

[0479] In a preferred embodiment, the pharmaceutical composition comprising docetaxel is formulated for parenteral administration.

[0480] A preferred embodiment of the treatment of cholangiocarcinoma using panobinostat in combination with docetaxel is to administer panobinostat orally and docetaxel as an injection or infusion.

[0481] In some embodiments, the clinical dose of panobinostat combined with docetaxel for the treatment of cholangiocarcinoma is generally 5 mg to 50 mg, more preferably 10 mg to 30 mg, administered daily or at least twice a week as defined above.

[0482] In some embodiments, the clinical dose of docetaxel combined with panobinostat for the treatment of cholangiocarcinoma is generally the same as the dose range currently used for docetaxel for other indications, for example, 20 mg / m2 per day. 2 Body surface area (BSA) up to 200 mg / m 2 Body surface area, preferably 40 mg / m 2 BSA to 75 mg / m 2 BSA.

[0483] According to the present invention, the drug substances disclosed herein (i.e., panobinostat and cytotoxic agents) can be in the form of free drugs or pharmaceutically acceptable salts, solvates or hydrates thereof. These salts, solvates and hydrates are described in detail in the art. According to the present invention, any suitable pharmaceutically acceptable salt, solvate or hydrate of the drug substances disclosed herein can be used to treat cholangiocarcinoma.

[0484] A preferred form of the drug substance is the form in which the drug substance is present in a drug product approved by a commercial regulatory agency.

[0485] The drugs may be administered simultaneously or sequentially. If the drugs are administered in the form of sequential administration, the time interval between drug administrations may range from minutes to days, depending on the nature of the drug substances and the clinical situation.

[0486] Thus, panobinostat and the additional cytotoxic agent may be used simultaneously, separately or sequentially. When used simultaneously, panobinostat and the additional cytotoxic agent are administered simultaneously, but may be administered by a single route or by separate routes (e.g., oral administration of a mixture or administration of two (or more) formulations simultaneously but by different routes (i.e., oral and intravenous)). When administered separately, panobinostat and the additional cytotoxic agent may be administered simultaneously or sequentially and / or may overlap in the time intervals between their administration. In some embodiments, these agents are administered together in a single formulation (mixture), e.g., panobinostat and dasatinib, panobinostat and topotecan, panobinostat and methotrexate, panobinostat and trametinib, panobinostat and BI 2536, panobinostat and combretastat A4, panobinostat and datolicoxib, panobinostat and dapoline, panobinostat and ispins, panobinostat and lumispivir, panobinostat and molibs, panobinostat and obaclava, panobinostat and peritinib, panobinostat and ilisimol, and panobinostat and triptolide.

[0487] In some embodiments of the invention, panobinostat and / or another cytotoxic agent is administered more than once, for example 2, 3, 4, 5, 6, 7, 8, 9 or 10 times (e.g. up to 20 times). This administration can be performed in a single (or each) cycle or in multiple cycles in total.

[0488] As mentioned herein, a "cycle" is a time period during which a specific treatment regimen is applied and is typically repeated to provide cyclical treatment. The treatment in each cycle may be the same or different (e.g., different doses, time intervals, etc. may be used). The duration of a cycle may be 7 to 30 days, for example, 14 days or 21 days for a cycle. In some embodiments, a cycle may be about 1 month to about 3 months. Multiple cycles may be used, for example, at least 2 cycles, 3 cycles, 4 cycles, or 5 cycles, for example, 6 cycles, 7 cycles, 8 cycles, 9 cycles, or 10 cycles (e.g., up to 8 cycles, 9 cycles, 10 cycles, or 20 cycles). As described above, within each cycle, panobinostat and / or another cytotoxic agent may be administered once or more than once.

[0489] If the combination drug therapy is administered separately or sequentially, the two or more different drugs (e.g., panobinostat and another cytotoxic agent) can be provided as a combination product, wherein these drugs are provided as separate formulations (e.g., ready-to-use formulations) for separate and / or sequential administration. For example, the combination product can include a kit or package containing the formulations and optional instructions for administration.

[0490] If the combination drug therapy is administered simultaneously, two or more different drugs (eg, panobinostat and another cytotoxic agent) may be administered together as a single pharmaceutical preparation in a so-called combination preparation.

[0491] Therefore, another embodiment of the present invention relates to a combined formulation (pharmaceutical composition) comprising panobinostat and one or more cytotoxic agents for the treatment of cholangiocarcinoma. In a preferred embodiment, one or more cytotoxic agents are selected from dasatinib, topotecan, methotrexate, trametinib, BI 2536, combretastatin A4, datolixib, dapoline, ilisimol, ispins, lumispi, molibs, obakla, pelitinib and triptolide and combinations thereof. This combined formulation can be easily prepared using known formulation techniques.

[0492] However, in some embodiments, different drugs may be administered simultaneously in separate forms, for example, separate tablets.

[0493] Thus, in a further embodiment, it will be seen that the present invention provides a kit as defined above comprising panobinostat and a cytotoxic agent, preferably for simultaneous, separate or sequential use to treat cholangiocarcinoma in a patient, wherein preferably the use is as defined above.

[0494] In a preferred embodiment, the cytotoxic agent is selected from bortezomib, BI 2536, carboplatin, cisplatin, combretastatin A4, datoxib, dapoline, dasatinib, doxorubicin, docetaxel, ilisimol, gemcitabine, ispinax, lumispivir, methotrexate, molibax, obaclava, pelitinib, SB-743921, topotecan, trametinib and triptolide, and combinations thereof.

[0495] In yet another preferred embodiment, the cytotoxic agent is selected from BI 2536, carboplatin, cisplatin, combretastatin A4, datocoxib, dapoline, dasatinib, doxorubicin, docetaxel, ilisimol, ispins, lumispi, methotrexate, molibs, obaclava, pelitinib, SB-743921, topotecan, trametinib and triptolide, and combinations thereof.

[0496] In another preferred embodiment, the additional cytotoxic agent is selected from doxorubicin, datolicoxib, SB-743921, trametinib, ilisimol, molibex, methotrexate, dapoline, topotecan, cisplatin, dasatinib, carboplatin and lumispivir.

[0497] In a further preferred embodiment, the additional cytotoxic agent is selected from carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib, datolicoxib, dapoline, ilisimol, ispins, lumispivir, molibs, obaclava, pelitinib, trametinib and triptolide, preferably selected from carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan and trametinib.

[0498] In addition to the above-mentioned drug substances and combinations for treating cholangiocarcinoma, the compositions, kits or treatment regimens of the present invention may include other drugs. These drugs may be other anti-cancer drugs or drugs known to be administered in cancer treatment regimens, such as other cytotoxic agents described herein.

[0499] In some embodiments of the present invention, before, simultaneously or after the treatment of the present invention, the subject (patient) can receive other treatments.For example, in some embodiments, the subject (patient) can be treated with radiotherapy and / or surgery according to methods known in the art.

[0500] Thus, in some embodiments, the methods of the invention may include the additional step of treating the subject with radiation therapy and / or surgery.Surgery may include resection of a cholangiocarcinoma tumor.

[0501] In some embodiments, the combination therapy of the present invention can be used as a second-line treatment, i.e., for subjects who have not responded well to gemcitabine-based therapy. Thus, in some embodiments, the subject to be treated has not responded well to gemcitabine-based therapy.

[0502] BSA (body surface area) can be calculated, for example, using the Mosteller formula (√([height (cm)×weight (kg)] / 3600)). If necessary, this can be calculated by using the conversion factor for an average adult (0.025 mg / kg=1 mg / m 2 ) was converted to mg / kg.

[0503] Preferred aspects of the invention are described in the examples, wherein one or more of the parameters or components used in the examples may be used as preferred features of the methods described above.

[0504] The invention will now be described in more detail in the following non-limiting examples with reference to the following drawings, in which:

[0505] Figure 1 Concentration response curves of panobinostat for seven cholangiocarcinoma cell lines are shown. A) Intrahepatic cholangiocarcinoma cell line. B) Extrahepatic cholangiocarcinoma cell line and KMCH-1 are combined cholangiocarcinoma and hepatoma cell lines. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0506] Figure 2 The effect of panobinostat (squares) and bortezomib (triangles) as single substance treatments and the combination of panobinostat with 1.3 nM bortezomib (circles) in the HuCC-T1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0507] Figure 3 The effect of panobinostat (squares) and carboplatin (triangles) as single substance treatments and the combination of panobinostat with 1000 nM carboplatin (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0508] Figure 4 The effect of panobinostat (squares) and cisplatin (triangles) as single substance treatments and the combination of panobinostat with 100 nM cisplatin (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0509] Figure 5 The effect of panobinostat (squares) and dasatinib (triangles) as single substance treatments and the combination of panobinostat with 5 nM dasatinib (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0510] Figure 6 The effect of panobinostat (squares) and doxorubicin (triangles) as single substance treatments and the combination of panobinostat with 87 nM or 100 nM doxorubicin (circles) in (A) HuCCT-1 cell line and (B) TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0511] Figure 7 The effect of panobinostat (squares) and gemcitabine (triangles) as single substance treatments and in combination with 12 nM or 1000 nM gemcitabine (circles) in (A) CC-SW-1 cell line and (B) TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0512] Figure 8 The effect of panobinostat (squares) and methotrexate (triangles) as single substance treatments and the combination of panobinostat with 24 nM methotrexate (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0513] Fig. 9 The effect of panobinostat (squares) and trametinib (triangles) as single substance treatments and combinations of panobinostat with 8.8 nM or 1000 nM trametinib (circles) in (A) HuCCT-1 cell line and (B) TFK-1 cell line are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0514] Fig.10 The effects of panobinostat (squares) and topotecan (triangles) as single substance treatments and the combination of panobinostat with 24 nM or 120 nM topotecan (circles) in (A) CC-SW-1 cell line and (C) TFK-1 cell line are shown. (B) The effects of topotecan (squares) and panobinostat (triangles) as single substance treatments and the combination of topotecan with 5.3 nM panobinostat (circles) in TFK-1 cell line are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0515] Fig.11 The effect of panobinostat (squares) and BI 2536 (triangles) as single substance treatments and the combination of panobinostat with 490 nM BI 2536 (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0516] Fig.12 The effect of panobinostat (squares) and triptolide (triangles) as single substance treatments and the combination of panobinostat with 30 nM triptolide (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0517] Fig.13 The effects of panobinostat (squares) and datolixibub (triangles) as single substance treatments and combinations of panobinostat with 80 nM or 2 nM datolixibub (circles) in (A) EGI-1 cell line and (B) TFK-1 cell line, respectively, are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0518] Fig.14 Shown are (A) the effect of panobinostat (squares) and dapolinet (triangles) as single substance treatments and the combination of panobinostat and 23 nM dapolinet (circles) in the TFK-1 cell line; and (B) the effect of dapolinet (squares) and panobinostat (triangles) as single substance treatments and the combination of daponatide and 5.3 nM dapolinet (circles) in the CC-SW-1 cell line. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0519] Fig.15 The effect of panobinostat (squares) and obaclava mesylate (triangles) as single substance treatments and the combination of panobinostat with 16 nM obaclava mesylate (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0520] Fig.16 The effect of panobinostat (squares) and SB-743921 (triangles) as single substance treatments and the combination of panobinostat with 6.4 nM SB-743921 (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0521] Fig.17 The effects of panobinostat (squares) and combretastat A4 (triangles) as single substance treatments and combinations of panobinostat with 1000 nM or 100 nM combretastat A4 (circles) in (A) EGI-1 cell line and (B) HuCC-T1 cell line, respectively, are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0522] Fig.18 The effect of panobinostat (squares) and ispinas (triangles) as single substance treatments and the combination of panobinostat with 75 nM ispinas (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0523] Fig.19 The effect of panobinostat (squares) and molibux (triangles) as single substance treatments and the combination of panobinostat with 1000 nM molibux (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0524] Fig. 20 The effect of panobinostat (squares) and lumibigostat (triangles) as single substance treatments and the combination of panobinostat with lumibigostat at 56 nM (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0525] Fig.21 The effect of panobinostat (squares) and peritinib (triangles) as single substance treatments and the combination of panobinostat 90 nM and peritinib (circles) in the TFK-1 cell line is shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0526] Fig. 22 (A) and (B) show the effects of ilisimo (squares) and panobinostat (triangles) as single substance treatments and the combination of ilisimo with 5.3nM or 14nM panobinostat (circles) in CC-SW-1 cell line and EGI-1 cell line, respectively. (C) and (D) show the effects of panobinostat (squares) and ilisimo (triangles) as single substance treatments and the combination of panobinostat with 7nM or 70nM ilisimo (circles) in HuCC-T1 cell line and TFK-1 cell line, respectively. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0527] Example

[0528] Experimental methods and analytical instructions for drug combinations

[0529] Various cholangiocarcinoma (CCA) cell lines were used for this drug screening. Table 1 provides details of the culture medium and cell numbers used for the experiments.

[0530] Table 1: Cell culture conditions of cholangiocarcinoma cell lines

[0531] Cell lines Cell culture medium Cells / well EGI-1 DMEM high glucose, 10% FCS, antibiotics 750 HuCC-T1 RPMI GlutaMAX, 10% FCS, antibiotics 750 TFK-1 RPMI GlutaMAX, 10% FCS, antibiotics 500 CC-SW-1 RPMI GlutaMAX, 10% FCS, antibiotics 500 KMBC DMEM high glucose, 10% FCS, antibiotics 750 CC-LP-1 RPMI GlutaMAX, 10% FCS, antibiotics 750 KMCH-1 RPMI GlutaMAX, 10% FCS, antibiotics 500

[0532] After trypsinization and counting, single cells were seeded into Greier 384-well tissue culture treated polystyrene plates (#781098) in 10 μL of appropriate culture medium (see Table 1 for exact cell numbers). The seeded cells were allowed to attach to the plate within 24 hours, and then an acoustic liquid dispenser (Labcyte Echo 550) was used to add the appropriate volume of compound to obtain a single drug concentration of 0.1 nM to 1000 nM ("dose response") in a total volume of 25 μL. 15 μL of appropriate culture medium was filled into the wells and incubated as above for 48 hours. After 48 hours of incubation, the cells were treated with 25 μL of 0.5× Cell TitreGlo Luminescent Activity Reagent. The cells were incubated in the dark for 10 minutes, and then the luminescence readings were read from the top using the automatic gain function on the Synergy Neo2 plate reader.

[0533] For panobinostat as a single substance treatment, seven cholangiocarcinoma cell lines were used. Panobinostat in combination with 23 other drugs was then tested on four cholangiocarcinoma cell lines. Based on the previously determined IC20 values ​​of the combination drugs tested individually on each of the four cell lines, a single concentration of the combination drug was added. The TFK-1 cell line is insensitive to some of the test drugs. If the IC20 value cannot be calculated, a dose of 100nM to 1000nM is selected for combination testing (applicable to carboplatin, cisplatin, gemcitabine, trametinib). For HuCCT-1 cells, trametinib was added at an IC44=8.8nM instead of IC20.

[0534] Data analysis

[0535] Cell viability from the Cell TitreGlo assay generated 576 dose-response curves for each cell line tested (24 for monotherapy using DMSO and 552 for combinations of drugs at IC20 concentrations).

[0536] Example 1: Effects of Panobinostat on Seven Cholangiocarcinoma Cell Lines

[0537] As described in detail in the Methods section above, panobinostat as a single substance treatment was tested on seven different cholangiocarcinoma cell lines compared to DMSO. These revealed that all analyzed cell lines showed a response to panobinostat (see Figure 1 ): Five cell lines showed an IC50 of approximately 100 nM, and two cell lines (CC-SW-1 and EGI-1) showed higher sensitivity to panobinostat with an IC50 of 12 nM or 36 nM (see IC50 values ​​in Table 2 and Figure 1 Dose response curve in ).

[0538] Although effective as a single agent, the efficacy of panobinostat is enhanced by combination with other cancer drugs (Examples 2 to 22) as described below.

[0539] Table 2: IC50 values ​​of panobinostat in seven cholangiocarcinoma cell lines

[0540] Cell lines Cell Type IC50(nM) CC-SW-1 Intrahepatic bile duct carcinoma 12 HuCC-T1 Intrahepatic bile duct carcinoma 90 EGI-1 Intrahepatic bile duct carcinoma 36 CC-LP-1 Intrahepatic bile duct carcinoma 81 TFK-1 Extrahepatic bile duct cancer 95 KMBC Extrahepatic bile duct cancer 88 KMCH-1 Cholangiocarcinoma and liver cancer combined 99

[0541] Example 2: Panobinostat combined with bortezomib for HuCCT-1 cholangiocarcinoma cells

[0542] HuCCT-1 cell line was treated with a combination of panobinostat and low-dose bortezomib (1.3 nM, single substance curve for IC20 dose, see Figure 2 For experimental details on combination drug testing, please refer to the Methods section above.

[0543] In fact, the combination of panobinostat and bortezomib was more effective, as highlighted by the reduction of the IC50 value of panobinostat from 90 nM as a single substance treatment to 51 nM when combined with 1.3 nM bortezomib. Therefore, it can be concluded that panobinostat combined with bortezomib showed a higher efficacy compared to single substance treatments.

[0544] Example 3: Panobinostat combined with carboplatin for TFK-1 cholangiocarcinoma cells

[0545] For experimental details of the combination drug testing, please refer to the above methods section. Interestingly, carboplatin ( Figure 3 , triangles) had no effect on the cell viability of TFK-1 cells. However, when carboplatin was added to panobinostat at a dose of 1000 nM ( Figure 3 , circles), the efficacy of panobinostat was increased, as shown by a decrease in the IC50 value from 70 nM to 26 nM ( Figure 3 ). Therefore, the combination of panobinostat and carboplatin is expected to show better therapeutic effect for cholangiocarcinoma than panobinostat alone.

[0546] Example 4: Combination of Panobinostat and Cisplatin for TFK-1 Cholangiocarcinoma Cells

[0547] Cisplatin as a single agent therapy Figure 4 , triangles) had no effect on TFK-1. However, the addition of 100 nM cisplatin to panobinostat (IC50 = 42 nM; Figure 4 , circles) showed higher efficacy than panobinostat alone (IC50 = 70 nM; squares), see Figure 4 For experimental details of the combination drug testing, please refer to the above methods section. Therefore, it is expected that the combination of panobinostat and cisplatin will have higher efficacy for cholangiocarcinoma treatment.

[0548] Example 5: Combination of Panobinostat and Dasatinib in TFK-1 Cholangiocarcinoma Cell Line

[0549] For experimental details on the combination drug testing, please refer to the above methods section. Figure 5 , triangles) showed only limited efficacy in TFK-1 cells. Interestingly, the addition of low-dose dasatinib (5 nM, single-substance curves for IC20 doses) augmented the effect of panobinostat. This was achieved by increasing the IC50 value from panobinostat alone ( Figure 5 , squares) moves down from 70 nM to the combined ( Figure 5 , circle) is illustrated by 39 nM. Therefore, from these results it is expected that the combination of panobinostat and dasatinib has a higher efficacy for cholangiocarcinoma treatment than panobinostat alone.

[0550] Example 6: Panobinostat combined with doxorubicin for HuCCT-1 cholangiocarcinoma cells and TFK-1 cholangiocarcinoma cells

[0551] For experimental details of the combination drug testing, please refer to the Methods section above. The HuCCT-1 cell line was sensitive to panobinostat and doxorubicin with IC50 values ​​of 90 nM and 158 nM, respectively. The combination of panobinostat and doxorubicin ( Figure 6 A, circles) showed a better response with an IC50 value of 44 nM, see Figure 6 A.

[0552] The TFK-1 cell line was sensitive to panobinostat and doxorubicin with IC50 values ​​of 70 nM and 123 nM, respectively. The combination of panobinostat and doxorubicin ( Figure 6 B, circles) showed a better response with an IC50 value of 40 nM, see Figure 6 B.

[0553] Therefore, the combination of panobinostat and doxorubicin is expected to be beneficial in cholangiocarcinoma treatment.

[0554] Example 7: Combination of Panobinostat and Gemcitabine in CC-SW-1 cholangiocarcinoma cell line and TFK-1 cholangiocarcinoma cell line Tie For experimental details on the combination drug testing, please refer to the above methods section. Figure 7 A, squares) and gemcitabine ( Figure 7 A, triangles) but were particularly sensitive to the combination of panobinostat and 12 nM gemcitabine ( Figure 7 A, circles) were sensitive to panobinostat alone, which had an IC50 value of 12 nM. The combination showed a lower IC50 value of 3 nM.

[0555] Interestingly, the TFK-1 cell line was insensitive to gemcitabine as a single agent. However, compared to panobinostat alone (IC50 = 70 nM, see Figure 7 B) Addition of gemcitabine (IC50 = 46 nM) to panobinostat increased efficacy in comparison.

[0556] Therefore, the addition of gemcitabine to panobinostat during cholangiocarcinoma treatment would increase the effect on cell viability.

[0557] Example 8: Panobinostat combined with methotrexate for TFK-1 cholangiocarcinoma cells

[0558] For experimental details on the combination drug testing, please refer to the Methods section above. Figure 8 , triangles) had only a minor effect on cell viability in TFK-1 cells. However, compared with panobinostat alone ( Figure 8 , squares, IC50 value of 70 nM) compared to the addition of 24 nM methotrexate to panobinostat ( Figure 8 , circles, IC50 value 37 nM) showed increased efficacy (see Figure 8 ). Similar effects are expected for the treatment of bile duct cancer.

[0559] Example 9: Panobinostat combined with trametinib in HuCCT-1 cholangiocarcinoma cell line and TFK-1 cholangiocarcinoma cell line Tie

[0560] Please refer to the above methods section for experimental details of the combination drug testing. Fig. 9 A and Fig. 9 B) showed only minor efficacy in both HuCCT-1 cells and TFK-1 cells. However, the addition of low-dose trametinib (8.8 nM) to panobinostat in HuCCT-1 cells increased the effect of panobinostat on cell viability. This was achieved by increasing the IC50 value from panobinostat alone ( Fig. 9 A, squares) shifted to 90 nM for the combined treatment (circles, Fig. 9 A) is shown as 42 nM.

[0561] Similarly, the combination of trametinib and panobinostat also showed a greater effect than panobinostat alone in TFK-1 cells by reducing the IC50 value from 70 nM to 31 nM ( Fig. 9 B).

[0562] Therefore, the combination therapy of panobinostat and trametinib is predicted to show a greater effect in the treatment of cholangiocarcinoma.

[0563] Example 10: Combination of Panobinostat and Topotecan in CC-SW-1 cholangiocarcinoma cell line and TFK-1 cholangiocarcinoma cell line Tie

[0564] For experimental details of the combination drug testing, please refer to the Methods section above. The CC-SW-1 cell line was sensitive to the combination of panobinostat and topotecan with IC50 values ​​of 12 nM and 54 nM, respectively. The combination of panobinostat with a fixed dose of topotecan (squares) was significantly more sensitive to panobinostat than panobinostat alone (squares). Fig.10 A, circles) showed a better response in CC-SW-1 cells, with an IC50 value of 5.0 nM, see Fig.10 A. In addition, the combination of topotecan with fixed-dose panobinostat ( Fig.10 B, circles) also showed a better response in CC-SW-1 cells, with an IC50 value of 37 nM, see Fig.10 B.

[0565] The TFK-1 cell line was also sensitive to the combination of panobinostat and topotecan, with IC50 values ​​of 70 nM and 373 nM, respectively. The combination of panobinostat with a fixed dose of topotecan ( Fig.10 C, circles) showed a better response in TFK-1 cells, with an IC50 value of 28 nM, see Fig.10 C.

[0566] Therefore, the combination of panobinostat and topotecan is expected to be beneficial in the treatment of cholangiocarcinoma.

[0567] Example 11: Combination of Panobinostat and BI 2536 in TFK-1 cholangiocarcinoma cells

[0568] For experimental details of the combination drug testing, please refer to the Methods section above. The TFK-1 cell line was sensitive to panobinostat and BI 2536 with IC50 values ​​of 70 nM and 80 nM, respectively. The combination of panobinostat and BI 2536 ( Fig.11 , circles) showed a better response in TFK-1 cells, with an IC50 value of 38 nM, see Fig.11 Therefore, the combination of panobinostat and BI 2536 is expected to be beneficial in cholangiocarcinoma treatment.

[0569] Example 12: Panobinostat combined with triptolide for TFK-1 cholangiocarcinoma cells

[0570] For experimental details of the combination drug testing, please refer to the Methods section above. The TFK-1 cell line was sensitive to panobinostat and triptolide with IC50 values ​​of 70 nM and 15 nM, respectively. The combination of panobinostat and triptolide ( Fig.12 , circles) showed a better response in TFK-1 cells, with an IC50 value of 64 nM, see Fig.12Therefore, the combination of panobinostat and triptolide is expected to be beneficial in cholangiocarcinoma treatment.

[0571] Example 13: Panobinostat combined with datoxib for EGI-1 cholangiocarcinoma cell line and TFK-1 cholangiocarcinoma cell line Tie

[0572] For experimental details of the combination drug testing, please refer to the Methods section above. The EGI-1 cell line was sensitive to panobinostat and datolicoxib with IC50 values ​​of 99 nM and 99 nM, respectively. The combination of panobinostat with a fixed dose of datolicoxib (squares) was significantly more sensitive to panobinostat than panobinostat alone (squares). Fig.13 A, circles) showed a better response in EGI-1 cells, with an IC50 value of 34 nM, see Fig.13 A.

[0573] The TFK-1 cell line was also sensitive to panobinostat and datolicoxib, with IC50 values ​​of 70 nM and 93 nM, respectively. The combination of panobinostat with a fixed dose of datolicoxib (squares) was significantly more sensitive to panobinostat than to panobinostat alone (squares). Fig.13 B, circles) showed a better response in TFK-1 cells, with an IC50 value of 30 nM, see Fig.13 B. Therefore, the combination of panobinostat and datolicoxib is expected to be beneficial in the treatment of cholangiocarcinoma.

[0574] Example 14: Combination of Panobinostat and Dapoline for TFK-1 cholangiocarcinoma cell line and CC-SW-1 cholangiocarcinoma cell line Tie

[0575] For experimental details of the combination drug testing, please refer to the Methods section above. The TFK-1 cell line was sensitive to panobinostat and datolicoxib with IC50 values ​​of 70 nM and 73 nM, respectively. The combination of panobinostat with a fixed dose of datolicoxib ( Fig.14 A, circles) showed a better response in TFK-1 cells, with an IC50 value of 48 nM, see Fig.14 A.

[0576] In addition, the CC-SW-1 cell line was sensitive to panobinostat and dapoline with IC50 values ​​of 12 nM and 8 nM, respectively. Fig.14 B, circles) showed a better response in TFK-1 cells, with a drug sensitivity score (DSS) of 34 for the combination compared to 17 for dapolinet alone (squares). Therefore, the combination of panobinostat and dapolinet is expected to be beneficial for cholangiocarcinoma treatment, see Fig.14 B.

[0577] Example 15: Panobinostat combined with ovaclazide mesylate for TFK-1 cholangiocarcinoma cells

[0578] For experimental details of the combination drug testing, please refer to the Methods section above. The TFK-1 cell line was sensitive to panobinostat and obaclava mesylate with IC50 values ​​of 70 nM and 1523 nM, respectively. The combination of panobinostat with a fixed dose of obaclava mesylate ( Fig.15 , circles) showed a better response in TFK-1 cells, with an IC50 value of 52 nM, see Fig.15 Therefore, the combination of panobinostat and abaclase mesylate is expected to be beneficial in cholangiocarcinoma treatment.

[0579] Example 16: Combination of Panobinostat and SB-743921 in TFK-1 cholangiocarcinoma cell line

[0580] For experimental details on combination drug testing, please refer to the Methods section above.

[0581] The TFK-1 cell line was also sensitive to panobinostat and SB-743921 with IC50 values ​​of 70 nM and 8 nM, respectively. The combination of panobinostat with a fixed dose of SB-743921 ( Fig.16 , circles) showed a better response in TFK-1 cells, with an IC50 value of 37 nM, see Fig.16 .

[0582] Therefore, the combination of panobinostat and SB-743921 is expected to be beneficial in cholangiocarcinoma treatment.

[0583] Example 17: Combination of Panobinostat and Combretastatin A4 in EGI-1 cholangiocarcinoma cells and HuCC-T1 cholangiocarcinoma cells

[0584] For experimental details of the combination drug testing, please refer to the Methods section above. The EGI-1 cell line was sensitive to panobinostat but not to combretastatin A4, with IC50 values ​​of 99 nM and >1000 nM, respectively. The combination of panobinostat with a fixed dose of combretastatin A4 ( Fig.17 A, circles) showed a better response in EGI-1 cells, with an IC50 value of 52 nM, see Fig.17 A.

[0585] The HuCC-T1 cell line was also sensitive to panobinostat and combretastatin A4, with IC50 values ​​of 90 nM and 11 nM, respectively. The combination of panobinostat with a fixed dose of combretastatin A4 ( Fig.17 B, circles) showed a better response in HuCC-T1 cells, with an IC50 value of 53 nM, see Fig.17 B.

[0586] Therefore, the combination of panobinostat and combretastatin A4 is expected to be beneficial in cholangiocarcinoma treatment.

[0587] Example 18: Combination of Panobinostat and Ispins for TFK-1 Cholangiocarcinoma Cells

[0588] Please refer to the Methods section above for experimental details of the combination drug testing. The TFK-1 cell line was sensitive to panobinostat and ispinas with IC50 values ​​of 70 nM and 11 nM, respectively. The combination of panobinostat with a fixed dose of ispinas (squares) was significantly more sensitive to panobinostat than panobinostat alone (squares). Fig.18 , circles) showed a better response in TFK-1 cells, with an IC50 value of 45 nM, see Fig.18 Therefore, the combination of panobinostat and ispinax is expected to be beneficial in cholangiocarcinoma treatment.

[0589] Example 19: Panobinostat combined with molibax in TFK-1 cholangiocarcinoma cells

[0590] Please refer to the Methods section above for experimental details of the combination drug testing. The TFK-1 cell line was sensitive to panobinostat and molibux with IC50 values ​​of 70 nM and 181 nM, respectively. The combination of panobinostat with a fixed dose of molibux ( Fig.19 , circles) showed a better response in HuCC-T1 cells, with an IC50 value of 26 nM, see Fig.19 Therefore, the combination of panobinostat and molibex is expected to be beneficial in cholangiocarcinoma treatment.

[0591] Example 20: Panobinostat combined with lumirastatin for TFK-1 cholangiocarcinoma cells

[0592] For experimental details of the combination drug testing, please refer to the Methods section above. The TFK-1 cell line was sensitive to panobinostat and lumispivir with IC50 values ​​of 70 nM and 21 nM, respectively. The combination of panobinostat with a fixed dose of lumispivir (squares) was significantly more sensitive to panobinostat than panobinostat alone (squares). Fig. 20 , circles) showed a better response in TFK-1 cells, with an IC50 value of 11 nM, see Fig. 20 Therefore, the combination of panobinostat and lumispivir is expected to be beneficial in cholangiocarcinoma treatment.

[0593] Example 21: Combination of Panobinostat and Peltinib in TFK-1 Cholangiocarcinoma Cells

[0594] For experimental details of the combination drug testing, please refer to the Methods section above. The TFK-1 cell line was sensitive to panobinostat and peritinib with IC50 values ​​of 70 nM and 0.08 nM, respectively. The combination of panobinostat with a fixed dose of peritinib (squares) was significantly more sensitive to panobinostat than to panobinostat alone (squares). Fig.21 , circles) showed a better response in TFK-1 cells, with an IC50 value of 34 nM, see Fig.21Therefore, the combination of panobinostat and peritinib is expected to be beneficial in cholangiocarcinoma treatment.

[0595] Example 22: Panobinostat combined with ilisimol for CC-SW-1 cholangiocarcinoma cell line and EGI-1 cholangiocarcinoma cell line line, HuCC-T1 cholangiocarcinoma cell line and TFK-1 cholangiocarcinoma cell line

[0596] For experimental details of the combination drug testing, please refer to the Methods section above. The CC-SW-1 cell line was sensitive to panobinostat and ilisimol with IC50 values ​​of 12 nM and 50 nM, respectively. The combination of panobinostat with a fixed dose of ilisimol (squares) was significantly more sensitive to panobinostat than ilisimol alone (squares). Fig. 22 A, circles) showed a better response in CC-SW-1 cells, with an IC50 value of 19 nM, see Fig. 22 A.

[0597] The EGI-1 cell line was also sensitive to panobinostat and ilisimol, with IC50 values ​​of 99 nM and 34 nM, respectively. The combination of panobinostat with a fixed dose of ilisimol (squares) showed a significant increase in the sensitivity of the cells to panobinostat and ilisimol compared to ilisimol alone (squares). Fig. 22 B, circles) showed a better response in EGI-1 cells, with an IC50 value of 19 nM, see Fig. 22 B.

[0598] The HuCC-T1 cell line was also sensitive to panobinostat and ilisimol, with IC50 values ​​of 90 nM and 9 nM, respectively. The combination of panobinostat with a fixed dose of ilisimol ( Fig. 22 C, circles) showed a better response in HuCC-T1 cells, with an IC50 value of 46 nM, see Fig. 22 C.

[0599] The TFK-1 cell line was also sensitive to panobinostat and ilisimol, with IC50 values ​​of 70 nM and 35 nM, respectively. The combination of panobinostat with a fixed dose of ilisimol ( Fig. 22 D, circles) showed a better response in TFK-1 cells, with an IC50 value of 24 nM, see Fig. 22 D. Therefore, the combination of panobinostat and ilisimol is expected to be beneficial in the treatment of cholangiocarcinoma.

[0600] Example 23: Capsules containing panobinostat and dasatinib

[0601] Panobinostat (99% purity) can be purchased from Shandong Sanrui Technology Co., Ltd., China. Alternatively, panobinostat lactate can be produced from panobinostat and lactic acid according to WO2007146716 (incorporated herein by reference). Dasatinib monohydrate (≥99.0% purity) can be purchased from Beijing Yibai Biotechnology Co., Ltd., China.

[0602] Capsules containing panobinostat and dasatinib were prepared as follows:

[0603] Components

[0604] Panobinostat lactate (equivalent to 15g Panobinostat)

[0605] Dasatinib monohydrate (equivalent to 50g dasatinib)

[0606] 1g magnesium stearate

[0607] 50g mannitol

[0608] Microcrystalline cellulose (up to 500 g)

[0609] The components were mixed by volume in a mixer and filled into 1000 hard gelatin capsules size 0. Each capsule contained 15 mg of panobinostat and 50 mg of dasatinib.

[0610] Example 24: Drug Product Comprising Two Different Drug Formulations

[0611] Panobinostat (99% purity) can be purchased from Shandong Sanrui Technology Co., Ltd., China. Alternatively, panobinostat lactate can be produced from panobinostat and lactic acid according to WO2007146716 (incorporated herein by reference).

[0612] Capsules similar to 15 mg Farydak (Novartis) were prepared.

[0613] The capsules are packed in blisters (6 capsules per blister).

[0614] Dasatinib monohydrate (purity ≥ 99.0%) can be purchased from Beijing Yibai Biotechnology Co., Ltd., China.

[0615] Tablets similar to 50 mg Sprycel (Bristol-Myers Squibb) were prepared.

[0616] Tablets are packaged in blisters (6 tablets per blister)

[0617] The blisters (5 tablet blisters and 5 capsule blisters) are packed in the medicine package together with the package insert.

[0618] Example 25: Panobinostat in combination with cytotoxic agents compared to panobinostat alone in normal cholangiocytes Reduced toxicity

[0619] The effects of various cytotoxic agents on panobinostat toxicity in normal cholangiocytes were examined. The H69 cell line (CVCL_8121) was used in the above experimental method to determine the IC50 value of panobinostat (primary drug) in these cells when combined with a cytotoxic agent (secondary drug) added to the cells at an IC20 concentration.

[0620] The effect of the secondary drug was quantified using the ΔIC50 measurement, which is calculated as the IC50 of panobinostat alone minus the IC50 of the panobinostat combination. A positive number indicates that the combination is more toxic than panobinostat alone. The greater the difference, the more toxic the combination is. A negative ΔIC50 indicates that the combination is less toxic than the monotherapy. The results are shown below:

[0621]

[0622] The results showed that the secondary drugs tested reduced the toxicity of panobinostat in normal cholangiocytes.

[0623] Example 26: Therapeutic Index of Panobinostat in Combination with Other Cytotoxic Agents Compared to Panobinostat Monotherapy

[0624] The therapeutic index of various panobinostat combination therapies was determined by comparing the effects of the combination and monotherapy (i.e., panobinostat alone) in normal cholangiocarcinoma cell lines (H-69 cell line) and various cholangiocarcinoma cell lines as described above. The IC50 values ​​of panobinostat (primary drug) in cells were determined using the above experimental method when used alone or in combination with a cytotoxic agent (secondary drug) added to the cells at an IC20 concentration.

[0625] The therapeutic index (TI) refers to the ratio of IC50 in normal cells to IC50 in cholangiocarcinoma cell lines. A TI greater than 1 indicates that the therapy is effective in reducing the viability of cholangiocarcinoma cells relative to normal cells. A high TI (e.g., 1.5 or higher) indicates that there is a large difference in efficacy between normal cells and cancer cells, i.e., the therapy shows a high selectivity for cancer cells relative to normal cells. A higher TI for the combination therapy than for the monotherapy indicates that the combination therapy is more selective for cancer cells than the monotherapy. The results are shown below (IC50 values ​​in nM):

[0626]

[0627]

[0628]

[0629]

[0630] These results suggest that the panobinostat combination therapies tested may be particularly effective against cholangiocarcinoma tumors that share features with the CC-SW-1 cell line. However, the data identified combination therapies that were effective in other cell lines. For example, the combination of trametinib with doxorubicin was particularly effective in the CC-SW-1 cell line. The combination of trametinib with topotecan was particularly effective in the EGI-1 cell line. The combination of trametinib with carboplatin was particularly effective in the HuCCT-1 cell line. The combination with dasatinib, methotrexate, datolicoxib, topotecan, and trametinib was particularly effective in the TFK-1 cell line.

[0631] Example 27: Panobinostat combination therapy modulating the IC50 of Panobinostat in cholangiocarcinoma cell lines

[0632] The experimental data described herein were used to identify cytotoxic agents that were particularly effective in promoting the effects of panobinostat in cholangiocarcinoma cells. Individual combinations were identified by determining the ΔIC50, where a higher positive ΔIC50 represents a more effective combination. The table below shows the absolute ΔIC50 (nM) and the relative change in percentage.

[0633]

[0634]

[0635]

[0636] The table below demonstrates that not all combinations were effective in all cell lines, ie some combinations showed negative effects on panobinostat toxicity in some cell lines, as indicated by negative ΔIC50 values.

[0637]

[0638] Example 28: Determination of the combination index of panobinostat and ilisimol

[0639] Data normalization and curve fitting

[0640] Cell viability from CTG assays X dose response curves were generated for each cell line tested (where Y bars were monotherapy with DMSO and Z bars were combinations of drugs at IC20 concentrations). The viability data for each plate were normalized to the average of eight replicates of 0.1% DMSO and seven replicates of 100uM benzethonium chloride (BzCl). BzCl served as a cell killing control, which explained the background signal from dead cells in the CTG luminescence assay. The raw luminescence data were normalized according to the following equation:

[0641]

[0642] The normalized data for each dose-response curve were then fitted using the drm function of the R package drc. This function fits the dose-response data using a four-parameter log-logistic curve to obtain the curve minimum, curve maximum, IC50 value, and slope value. In cases where the log-logistic curve could not fit the data, a logistic curve was used. The IC50 corresponds to the concentration between the curve maximum and the curve minimum calculated at 50% response and is therefore a relative IC50 value.

[0643] Log-logistic function:

[0644] Logical functions:

[0645] Synergy score calculation

[0646] According to the Loewe summation method, Bliss independence method and zero interaction potential (ZIP) method, the synergy score of the monotherapy dose response relative to the combined dose response of each drug combination is calculated. These methods calculate the predicted response based on the monotherapy response of the drugs used in the combination. The measured response of each combination is then subtracted from these predicted responses to generate a synergy score for each tested concentration; a positive score indicates synergy and a negative score indicates antagonism.

[0647] Loewe collaboration

[0648] The Loewe synergy value was calculated using the explicit method. For each drug in the combination, the predicted response was calculated from the response of that drug alone at a dose equal to the sum of the doses of the two drugs in the combination. The predicted responses for each drug were then averaged and the observed value at the measured concentration was subtracted from this average to produce a synergy score.

[0649]

[0650] in is monotherapy with drug 1 at a concentration of x 1 Drug 1 and concentration x 2 The response calculated by curve fitting under the sum of the concentrations of drug 2, is the response calculated using curve fitting of drug 2 monotherapy at the same sum of concentrations of drug 1 and drug 2, are drug 1 and drug 2 at their respective doses x 1 and x 2 The measured response of the combination of or When is >100 or <0, they are set to 100 and 0 respectively.

[0651] The Loewe additivity model is preferred when drugs used in combination target the same pathway, as they are expected to have additive effects.

[0652] Bliss Collaboration

[0653] The predicted responses generated using the Bliss model were calculated by multiplying the monotherapy responses of each drug at each tested concentration in the combination. The measured responses at these concentrations were then subtracted from these predicted values ​​to generate a synergy score.

[0654]

[0655] in is from drug 1 monotherapy at dose x 1 The curve fitting response is as follows, is from drug 2 monotherapy at dose x 2 The curve fitting response is as follows, are drug 1 and drug 2 at their respective doses x 1 and x 2 The measured response of the combination under or When either or both of them are >100 or <0, they are set to 100 and 0, respectively.

[0656] The Bliss independence model is preferred when the drugs used in combination target different pathways because they are expected to have independent effects.

[0657] ZIP Collaboration

[0658] The predicted response generated using the ZIP model was calculated according to the above-mentioned Bliss method. The observed combined response was fitted to a log-logistic function, and the curve maximum was set to the corresponding response of the IC20 drug monotherapy at the relevant dose. These fitted combined values ​​were then subtracted from the predicted response to generate a synergy score.

[0659]

[0660] in is from drug 1 monotherapy at dose x 1 The curve fitting response is as follows, is from drug 2 monotherapy at dose x 2 The curve fitting response is as follows, is the response calculated using a curve fit of drug 1 with a fixed dose of drug 2, where the upper parameter is set to drug 2 monotherapy at dose x 2 The following reaction. or When either or both of them are >100 or <0, they are set to 100 and 0, respectively.

[0661] The ZIP model was created to integrate the Bliss model and the Loewe model.

[0662] The combination index of panobinostat and ilisimol was determined as described herein. The table below shows that the combination showed synergy in three cell lines, ie, a combination index of less than 1.

[0663] Panobinostat concentration (nM) Elisemol concentration (nM) Cell lines Combined Index 0.1 20 CCSW1 0.3 1 20 CCSW1 0.6 3 20 CCSW1 0.7 10 20 CCSW1 0.7 0.1 20 EGI1 0.5 1 20 EGI1 0.5 3 20 EGI1 0.5 10 20 EGI1 0.7 30 20 EGI1 0.7 100 20 EGI1 0.6 0.1 7 HuCCT 0.4 1 7 HuCCT 0.5 3 7 HuCCT 0.6 10 7 HuCCT 0.7

[0664] Example 29: Genomic sequences of cell lines

[0665] Whole genome sequencing was performed on the cell lines used herein to identify genetic markers (mutations) that are specific to the cell lines and that are expected to be present in cholangiocarcinoma tumors. The following table shows markers associated with predictive, prognostic, diagnostic and susceptibility biomarkers in the CIViC database and the Cancer Biomarker Database, coding mutations found in known cancer mutation hotspots that are predicted to be cancer driver mutations or selected as pathogenic mutations and coding mutations found in oncogenes or tumor suppressor genes in the cell lines.

[0666]

[0667]

[0668]

[0669] Example 30: Mouse xenograft studies

[0670] Cell culture

[0671] Normal human cholangiocarcinoma cell line (H69) and various human cholangiocarcinoma cell lines (HuCCT, CC-SW1, EGI-1, and TFK-1) were cultured according to standard conditions.

[0672] Mouse experiments

[0673] All mouse experiments were performed according to protocols approved by the Norwegian Ethical Committee for the Use of Animals in Research. Animals were housed in cages with a temperature-controlled environment. Animals had free access to standard chow and water. The light / dark cycle was 12 h / 12 ​​h. Cell suspensions were injected subcutaneously into nude mice.

[0674] Ten days after administration of the cell suspension, tumor growth was confirmed. The mice were divided into 5 groups (10 animals per group); the first group was not treated with active treatment, the second group was treated with drug A, the third group was treated with drug B, the fourth group was treated with a combination of drug A and drug B, and the fifth group was treated with gemcitabine-based combination therapy. All animals had free access to feed and water.

[0675] If the drug is a regulatory approved drug, it is administered in the same dose (per kg) and dose frequency and in the same manner as it is clinically used to treat other cancer diseases. The highest dose and most frequent administration are used. If the drug is an experimental drug (i.e., a currently unapproved drug), it is administered in the same dose (per kg) and dose frequency and in the same manner as it is used in the prior art literature to treat cancer.

[0676] Tumor volume was measured weekly throughout the treatment period. Some mice were examined by ultrasound and / or magnetic resonance imaging (MRI) during the treatment period to observe tumor growth. Mice were anesthetized and sacrificed according to standard methods after 50 days. Tumors were removed, weighed and stored in a refrigerator for further analysis.

[0677] The results are expected to show that certain drug combinations are highly effective for the treatment of human cholangiocarcinoma in a xenograft nude mouse model. The in vivo efficacy is expected to correlate closely with the in vitro cell line efficacy.

[0678] Example 31: Clinical regimen using combination therapy of drug A and drug B as second-line therapy in patients with cholangiocarcinoma

[0679] Single-arm, open-label, non-randomized, exploratory, multicenter pilot study. Drug A and Drug B are drugs with regulatory approval for other cancer indications.

[0680] 30 participants

[0681] Inclusion criteria:

[0682] Patients diagnosed with cholangiocarcinoma by histology or cytology

[0683] Radiographically measurable disease (according to RECIST v1.1 criteria)

[0684] Patients previously treated with gemcitabine-based first-line therapy

[0685] Age: 18 to 80 years old, male or female

[0686] Women taking birth control pills (if relevant)

[0687] Exclusion criteria:

[0688] · Lactating or pregnant women

[0689] Severe heart dysfunction

[0690] Hypertension (systolic blood pressure ≥150 mmHg or diastolic blood pressure ≥100 mmHg)

[0691] Hepatitis C positive and / or human immunodeficiency virus (HIV) positive and / or novel coronavirus infection (Covid-19) positive

[0692] Primary sclerosing cholangitis and / or inflammatory bowel disease and / or autoimmune disease

[0693] Active drug treatment of systemic infections.

[0694] · History of allergy or serious adverse events to the combined drug or drugs with the same mechanism of action as the combined drug.

[0695] ·A history of substance abuse, including alcohol abuse and / or drug abuse.

[0696] Insufficient organ function

[0697] Absolute neutrophil count (ANC) <1,000 / mm 3 〔1.0×10 9 / L〕

[0698] ο Platelets <75,000 / mm 3 〔75×10 9 / L〕

[0699] oHemoglobin <109.0 g / dL

[0700] Total bilirubin > 1.5 × ULN

[0701] In the presence of liver metastasis, aspartate aminotransferase / aspartate aminotransferase / GOT (AST / SGOT) and alanine aminotransferase / glutamic acid aminotransferase / GPT (ALT / SGPT)>2.5×ULN (AST and ALT)>5×upper limit of normal (ULN)

[0702] Serum creatinine >1.5×ULN, calculated or measured creatinine clearance <45 mL / min

[0703] οInorganic phosphorus exceeds normal range

[0704] Serum total calcium and serum ionized calcium outside the normal range

[0705] Other protocol-defined inclusion / exclusion criteria may apply

[0706] Drug administration

[0707] The drugs are administered alone at 50% of the highest approved acceptable dose for the treatment of other forms of cancer. Each drug is administered in the same manner and at the same frequency as the drug for other indications. The two drugs are preferably administered together.

[0708] Duration

[0709] Each patient was dosed for up to 24 months.

[0710] Outcome Measures

[0711] Main objectives:

[0712] Objective response rate (ORR) [time frame: up to 24 months]

[0713] Defined as the proportion of participants in each cohort who achieved a complete response (CR) or partial response (PR) according to Response Evaluation Criteria in Solid Tumors, version 1.1 (RECIST v1.1).

[0714] Secondary Objectives:

[0715] Progression-free survival (PFS) [time frame: up to 24 months]

[0716] Defined as the time from the first dose until disease progression (according to RECIST v1.1 criteria) or death (whichever comes first) in each cohort. Median progression-free survival.

[0717] Duration of response (DOR) [Time frame: up to 24 months]

[0718] It was defined as the time from the date of the first assessment of CR or PR until the date of first disease progression (according to RECIST v1.1 criteria) or death (whichever came first) in each cohort.

[0719] Best overall efficacy [time frame: up to 24 months]

[0720] The best overall response will be summarized by the proportion of patients with the best overall response of PR, CR, stable disease (SD), or PD.

[0721] Disease control rate (DCR) [time frame: up to 24 months]

[0722] Defined as the proportion of participants who achieved a best overall response of CR, PR, or stable disease according to RECIST v1.1 criteria.

[0723] Overall survival (OS) [time frame: up to 24 months]

[0724] It was defined as the time from the first dose of study drug to death from any cause in each cohort.

[0725] Median overall survival.

[0726] Number of treatment-related adverse events [time frame: up to 24 months]

[0727] First reported adverse events and serious adverse events, type and frequency, or exacerbation of pre-existing events after first dose of study drug / treatment.

[0728] Quality of Life - Quality of Life Analysis. See: (https: / / www.eortc.org / app / uploads / sites / 2 / 2018 / 08 / Specimen-QLQ-C30-English.pdf)

[0729] Example 32: Combination therapy of drug A and drug B and gemcitabine and cisplatin in patients with cholangiocarcinoma Clinical Program

[0730] A double-arm, double-blind, randomized, multicenter phase III clinical study.

[0731] Drug X and Drug Y are drugs that have regulatory approval for other cancer indications.

[0732] 80 participants.

[0733] Arm A: Combination therapy with drug A and drug B (40 participants)

[0734] Arm B: Gemcitabine (1000 mg / m 2 ) and cisplatin (25 mg / m 2 ).

[0735] Inclusion criteria:

[0736] Patients diagnosed with cholangiocarcinoma by histology or cytology

[0737] Radiographically measurable disease (according to RECIST v1.1 criteria)

[0738] Age: 18 to 80 years old, male or female

[0739] Women taking birth control pills (if relevant)

[0740] Exclusion criteria:

[0741] · Lactating or pregnant women

[0742] Severe heart dysfunction

[0743] Hypertension (systolic blood pressure ≥150 mmHg or diastolic blood pressure ≥100 mmHg)

[0744] Hepatitis C positive and / or human immunodeficiency virus (HIV) positive and / or novel coronavirus infection (Covid-19) positive

[0745] Primary sclerosing cholangitis and / or inflammatory bowel disease and / or autoimmune disease

[0746] Active drug treatment of systemic infections.

[0747] · History of allergy or serious adverse events to the combined drug or drugs with the same mechanism of action as the combined drug.

[0748] ·A history of substance abuse, including alcohol and drug abuse.

[0749] Insufficient organ function

[0750] Absolute neutrophil count (ANC) <1,000 / mm 3 〔1.0×10 9 / L〕

[0751] ο Platelets <75,000 / mm 3 〔75×10 9 / L〕

[0752] oHemoglobin <109.0 g / dL

[0753] Total bilirubin > 1.5 × ULN

[0754] In the presence of liver metastasis, aspartate aminotransferase / aspartate aminotransferase / GOT (AST / SGOT) and alanine aminotransferase / glutamic acid aminotransferase / GPT (ALT / SGPT)>2.5×ULN (AST and ALT)>5×upper limit of normal (ULN)

[0755] Serum creatinine >1.5×ULN, calculated or measured creatinine clearance <45 mL / min

[0756] οInorganic phosphorus exceeds normal range

[0757] Serum total calcium and serum ionized calcium outside the normal range

[0758] Other protocol-defined inclusion / exclusion criteria may apply

[0759] Drug administration

[0760] The results of the exploratory studies form the basis for the dose of the combination drug. There is no relevant guidance. The combination drug is administered alone at 50% of the highest approved acceptable dose for the treatment of other forms of cancer. Each drug is administered in the same manner and at the same frequency as the drug for other indications. The two drugs are preferably administered together.

[0761] Duration

[0762] Each patient was dosed for 24 months.

[0763] Outcome Measures

[0764] Main objectives:

[0765] Objective response rate (ORR) [time frame: up to 24 months]

[0766] Defined as the proportion of participants in each cohort who achieved a complete response (CR) or partial response (PR) according to Response Evaluation Criteria in Solid Tumors, version 1.1 (RECIST v1.1).

[0767] Secondary Objectives:

[0768] Progression-free survival (PFS) [time frame: up to 24 months]

[0769] Defined as the time from the first dose until disease progression (according to RECIST v1.1 criteria) or death (whichever comes first) in each cohort. Median progression-free survival.

[0770] Duration of response (DOR) [Time frame: up to 24 months]

[0771] It was defined as the time from the date of the first assessment of CR or PR until the date of first disease progression (according to RECIST v1.1 criteria) or death (whichever came first) in each cohort.

[0772] Best overall efficacy [time frame: up to 24 months]

[0773] The best overall response will be summarized by the proportion of patients with the best overall response of PR, CR, stable disease (SD), or PD.

[0774] Disease control rate (DCR) [time frame: up to 24 months]

[0775] Defined as the proportion of participants who achieved the best overall response of CR, PR, or stable disease according to RECIST v1.1 criteria.

[0776] Overall survival (OS) [time frame: up to 24 months]

[0777] Defined as the time from the first dose of study drug to death from any cause in each cohort. Median overall survival.

[0778] Number of treatment-related adverse events [time frame: up to 24 months]

[0779] First reported adverse events and serious adverse events, type and frequency, or exacerbation of pre-existing events after first dose of study drug / treatment.

[0780] Quality of Life - Quality of Life Analysis. See: (https: / / www.eortc.org / app / uploads / sites / 2 / 2018 / 08 / Specimen-QLQ-C30-English.pdf)

[0781] Example 33: Reference Example - Analysis of the effect of a combination therapy comprising gemcitabine and cisplatin

[0782] The combination of gemcitabine and cisplatin is currently a common treatment for cholangiocarcinoma (see https: / / www.legemiddelhandboka.no / T2.2.1.4 / Galleveiscancer and Juan Valle et al., Annals of Oncology 25:391–398, 2014).

[0783] The combination was tested using the experimental method described above. The therapeutic index results for the combination using gemcitabine as the primary drug are shown below.

[0784]

[0785] Gemcitabine monotherapy is a preferred treatment for cholangiocarcinoma. Gemcitabine monotherapy data in CC-SW-1, EGI-1, HuCCT, and TFK-1 showed that the IC50 values ​​of normal cells were much lower than the IC50 values ​​of all cholangiocarcinoma cell lines. The therapeutic index was 0.4, 0.4, 0.4, and 0.2, respectively. This suggests that the administration of gemcitabine is not a good treatment for cholangiocarcinoma.

[0786] The drug combination of gemcitabine plus cisplatin is also a preferred clinical treatment for cholangiocarcinoma. The combined index data for CC-SW-1 cell lines and TFK-1 cell lines showed a certain improvement in the therapeutic index. However, for the EGI-1 cell line, the addition of cisplatin to gemcitabine destroyed the effect of gemcitabine. This combination had no therapeutic effect on the HuCCT cell line.

[0787] Example 34: Panobinostat combination therapy showing synergy in at least one cholangiocarcinoma cell line

[0788] The table below shows that panobinostat combination therapy exhibited synergistic effects in at least one cholangiocarcinoma cell line.

[0789]

[0790] In summary, the inventors have conducted extensive testing of anticancer drugs and combinations thereof. In this regard, there are currently thousands of known compounds with some reported activity against one or more forms of cancer. In order to obtain the results described herein, the inventors first selected a suitable compound library comprising 384 compounds. Such a compound library will produce more than 120,000 different combinations comprising two substances. By extensively testing single compounds and compound combinations, the inventors have identified 20 combinations that show good efficacy to at least one of the cell lines tested herein. This number is approximately 0.02% of the theoretical number of combinations based on the initial selection of 384 different compounds.

Claims

1. Use of panobinostat or a pharmaceutically acceptable salt thereof and trametinib or a pharmaceutically acceptable salt thereof in the preparation of a product for treating cholangiocarcinoma.

2. The use according to claim 1, wherein the panobinostat or a pharmaceutically acceptable salt thereof and trametinib or a pharmaceutically acceptable salt thereof are used or administered separately, simultaneously or sequentially.

3. The use according to claim 1 or 2, wherein the bile duct cancer is intrahepatic bile duct cancer. The use according to claim 1 or 2, wherein the bile duct cancer is extrahepatic bile duct cancer.

5. The use according to claim 1 or 2, wherein the panobinostat or a pharmaceutically acceptable salt thereof is provided in a pharmaceutical composition together with a pharmaceutically acceptable excipient.

6. The use according to claim 5, wherein the pharmaceutical composition is formulated for oral administration.

7. The use according to claim 5, wherein the pharmaceutical composition is in the form of a tablet or a capsule.

8. The use according to claim 5, wherein the pharmaceutical composition comprises trametinib or a pharmaceutically acceptable salt thereof.

9. The use according to claim 1 or 2, wherein the trametinib or a pharmaceutically acceptable salt thereof is formulated for oral administration.

10. The use according to claim 9, wherein the trametinib or a pharmaceutically acceptable salt thereof is in the form of a tablet or a capsule.

11. Use of a kit comprising panobinostat or a pharmaceutically acceptable salt thereof and trametinib or a pharmaceutically acceptable salt thereof in the preparation of a medicament for simultaneous, separate or sequential use to treat cholangiocarcinoma in a subject.

12. The use according to claim 11, wherein the bile duct cancer is intrahepatic bile duct cancer.

13. The use according to claim 11, wherein the cholangiocarcinoma is extrahepatic cholangiocarcinoma.

14. Use of a pharmaceutical composition of panobinostat or a pharmaceutically acceptable salt thereof and trametinib or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cholangiocarcinoma in a subject.

15. Use of a combination product of panobinostat or a pharmaceutically acceptable salt thereof and trametinib or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cholangiocarcinoma in a subject.

16. The use according to claim 15, wherein the panobinostat or a pharmaceutically acceptable salt thereof and trametinib or a pharmaceutically acceptable salt thereof are for separate, simultaneous or sequential use or administration.

17. The use according to claim 14 or 15, wherein the bile duct cancer is intrahepatic bile duct cancer.

18. The use according to claim 14 or 15, wherein the bile duct cancer is extrahepatic bile duct cancer.

19. The use according to claim 14 or 15, wherein the medicament comprises a pharmaceutically acceptable excipient.

20. The use according to claim 14 or 15, wherein the medicament is formulated for oral administration.

21. The use according to claim 14 or 15, wherein the medicament is in the form of a tablet or a capsule.

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