Combination therapy containing panobinostat for the treatment of cholangiocarcinoma

Abstract

The present invention relates to compositions and methods for the treatment of cholangiocarcinoma, and in particular to combination therapies comprising panobinostat compositions in combination with other cytotoxic agents, such as agents that enhance the effects of panobinostat, for use in the treatment of cholangiocarcinoma. Pharmaceutical compositions comprising panobinostat and other cytotoxic agents are also provided.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to compositions and methods for the treatment of cholangiocarcinoma. More particularly, the present invention relates to combination therapies comprising panobinostat compositions in combination with other cytotoxic agents, such as agents that enhance the effects of panobinostat, for use in the treatment of cholangiocarcinoma, and methods for treating cholangiocarcinoma by administering panobinostat in combination with other cytotoxic agents, such as agents that enhance the effects of panobinostat. [Background technology]

[0002] Background of the Invention There are over 100 forms of cancer that originate from specific cell types in various organs and 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, gene mutation burden, and transforming oncogenic mutations. One such example is breast cancer, which is further classified according to the expression of estrogen receptors, progesterone receptors, and HER2 receptors. Furthermore, triple-negative breast cancer does not express any of the above receptors.

[0003] For almost all forms of cancer, the prognosis is much better if the tumor is diagnosed at an early stage in the disease progression and the cancer is also grouped according to the stage of its progression. Different forms and stages of cancer generally have different treatment protocols.

[0004] For any given diagnosis, cancer treatments are further divided into first-line, second-line, and third-line, provided that treatment is based on established and available therapeutic regimens. Preferred treatments for various forms of cancer may vary slightly from country to country.

[0005] Cholangiocarcinoma (CCA, also known as biliary tract cancer) is a rare primary malignant tumor in Europe and North America, but is more common in Asian countries (Boris Blechacz: Cholangiocarcinoma: Current Knowledge and New Developments in Gut Liver. 2017 Jan; 11(1): 13-26).

[0006] In cholangiocarcinoma, cancer cells arise in the bile duct, either intrahepatically or extrahepatically. Therefore, CCA can be divided into intrahepatic and extrahepatic CCA. Extrahepatic CCA, accounting for 60–80% of CCAs, is further divided into perihilar CCA and distal CCA. The primary treatment for cholangiocarcinoma in Norway is surgery. However, 70–80% of extrahepatic CCAs are not candidates for curative resection. Radiation therapy is a valuable addition to treatment protocols. If patients have metastatic cholangiocarcinoma, drug treatment is generally gemcitabine combined with oxaliplatin, capecitabine, or cisplatin.

[0007] In recent years, various clinical studies for treating cholangiocarcinoma with multiple drugs and multiple combination drugs have been reported in the 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 peptide combinations for use in immunotherapy against gallbladder and bile duct cancers and other cancers.

[0009] WO2017 / 037299 provides a method for treating cancer of the biliary tract, such as cholangiocarcinoma, by administering a therapeutically effective amount of valitinib.

[0010] WO2008 / 023947 describes a pharmaceutical composition for inhibiting the growth or metastasis of cholangiocarcinoma, comprising a LICAM activity inhibitor or expression suppressor, and a treatment method using the composition. Summary of the Invention [Problem to be solved by the invention]

[0011] However, despite the development of novel treatments, cholangiocarcinoma is still considered a devastating malignancy with poor response to chemotherapy and fatal resistant complications.

[0012] The prognosis for patients with cholangiocarcinoma is generally very poor, and the clinical value of drug treatment in cholangiocarcinoma is limited. The 5-year survival rate is less than 5% and 0% when the tumor is inoperable. The median survival time is 12 months. Therefore, there is an urgent medical need for improved treatments. [Means for solving the problem]

[0013] Summary of the Invention In the research leading to this invention, the inventors selected over 380 known anticancer-related active pharmaceutical ingredients (e.g., cytotoxic agents) to extensively evaluate their effects alone and in combination on several cholangiocarcinoma cell lines. After rounds of selection based on known properties, such as activity against cholangiocarcinoma cell lines and efficacy at low doses of the combined substances, benign side effects, and known mechanisms of action, the inventors found that panobinostat was particularly effective against both intrahepatic and extrahepatic cholangiocarcinoma cell lines. This was particularly surprising given that only a small proportion of the drugs and drug combinations tested were active in cellular assays.

[0014] Furthermore, the present inventors have determined that several select drug substances (eg, cytotoxic agents) can enhance the anticancer activity of panobinostat against one or more cholangiocarcinoma cell lines.

[0015] Thus, in its broadest scope, the present invention includes a method of treating cholangiocarcinoma in a subject, 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 or a pharmaceutically acceptable salt thereof that enhances (i.e., potentiates) the therapeutic effect of panobinostat, wherein the cytotoxic agent is administered separately, simultaneously, or sequentially with the therapeutically effective amount of panobinostat or a pharmaceutically acceptable salt thereof.

[0016] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, and a cytotoxic agent, or a pharmaceutically acceptable salt thereof, that enhances (i.e., potentiates) the therapeutic effect of panobinostat, for use in treating cholangiocarcinoma in a subject.

[0017] In other embodiments, 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, or a pharmaceutically acceptable salt thereof, that enhances (i.e., potentiates) the therapeutic effect of panobinostat.

[0018] In a further embodiment, the present invention provides panobinostat or a pharmaceutically acceptable salt thereof for use in treating cholangiocarcinoma in a subject as a combination product with a cytotoxic agent for separate, simultaneous, or sequential use or administration to the subject. In certain embodiments, panobinostat or a pharmaceutically acceptable salt thereof can be formulated with the cytotoxic agent to provide a combined preparation, e.g., a pharmaceutical composition, comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent.

[0019] The present invention also provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with a cytotoxic agent for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject. In some embodiments, panobinostat or a pharmaceutically acceptable salt thereof can be formulated with the cytotoxic agent to provide a combination drug, e.g., a pharmaceutical composition, comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent.

[0020] Detailed Description of the Invention 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 can be prepared as described in WO 02 / 22577, which is incorporated herein by reference. References herein to panobinostat include salts thereof. [ka]

[0021] Pharmaceutically acceptable salts include pharmaceutically acceptable base addition salts and acid addition salts, such as metal salts, such as alkali and alkaline earth metal salts, ammonium salts, organic amine addition salts, 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 alkylsulfonates, arylsulfonates, 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 salts with morpholine and piperidine. Examples of amino acid addition salts are salts with glycine, phenylalanine, glutamic acid, and lysine. Sulfonates include mesylates, tosylates, and benzenesulfonates.

[0022] Preferred salts include organic acid addition salts such as alkylsulfonates, arylsulfonates, acetates, maleates, fumarates, tartrates, citrates and lactates, with the lactate salts being particularly preferred.

[0023] The above list of pharmaceutically acceptable salts applies to all drug substances described herein (eg, panobinostat and the cytotoxic agents described below) unless otherwise specified.

[0024] As used herein, "pharmaceutically acceptable" refers to a component that is compatible with other components used in the method or use of the present invention and is physiologically tolerable to the recipient.

[0025] "Choledochocarcinoma" or "CCA" refers to cholangiocarcinoma, which can be intrahepatic or extrahepatic (can be perihilar and distal). More than 90% of CCA is adenocarcinoma. In some embodiments, the CCA to be treated is metastatic CCA. In some embodiments, the CCA to be treated is intrahepatic CCA. In some embodiments, the CCA to be treated is extrahepatic CCA.

[0026] As detailed in the Examples, the inventors have determined that the combination therapies of the present invention have different effects on different cell lines. In this regard, the cell lines are derived from individual tumors and can be considered to represent different forms of CCA. For example, each cell line can have one or more characteristics commonly found in CCA tumors, such as one or more genetic markers, growth rate, cell morphology, or a combination thereof. Thus, the combination therapies disclosed herein that are particularly effective at inhibiting the growth or killing of a particular cell line are particularly useful for treating CCA tumors that have one or more characteristics associated with the CCA cell line, such as one or more genetic markers (e.g., mutations), growth rate, and / or cell morphology, e.g., one or more characteristics specific to the CCA cell line.

[0027] For example, the EGI-1 (CVCL_1193) and TFK-1 (CVCL_2214) cell lines were derived from explants of extrahepatic CCA tumors in male subjects (Shimizu et al. Int. J. Cancer 52:252-260 (1992), incorporated herein by reference). Thus, in certain embodiments, the combination therapies disclosed herein can be used to treat subjects with CCA tumors (e.g., extrahepatic CCA tumors) having one or more characteristics specific to the EGI-1 and / or TFK-1 cell lines, e.g., one or more genetic markers, growth rate, and / or cell morphology.

[0028] The CC-SW-1 cell line is derived from an explant of an intrahepatic CCA tumor of a female subject. Thus, in some embodiments, the combination therapy disclosed herein can be used to treat a subject with a CCA tumor (e.g., an intrahepatic CCA tumor) that has one or more characteristics specific to the CC-SW-1 cell line, such as one or more gene markers, growth rate, and / or cell morphology.

[0029] The HuCC-T1 cell line was derived from the ascites of a male subject with metastatic intrahepatic CCA tumors. Thus, in certain embodiments, the combination therapy disclosed herein can be used to treat a subject with an intrahepatic bile duct tumor (e.g., a subject with an intrahepatic CCA tumor, e.g., a metastatic intrahepatic bile duct tumor) that has one or more characteristics specific to the HuCC-T1 cell line, such as one or more gene markers, growth rate, and / or cell morphology.

[0030] The Examples section describes which combination therapies are effective in each cell line and therefore which treatments may be effective in treating CCA tumors as defined above. In a representative example, Examples 26 and 27 demonstrate that trametinib and doxorubicin are particularly effective in enhancing the effects of panobinostat in the CC-SW-1 cell line. Thus, in certain embodiments, the present invention provides combination therapies of panobinostat and trametinib or doxorubicin (e.g., as defined herein, including salts thereof, etc.) for use in treating subjects with CCA tumors (e.g., intrahepatic CCA tumors) 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.

[0031] Some combination therapies are effective against more than one cell line; for example, trametinib enhances the effects of panobinostat in CC-SW-1, EGI-1, HuCC-T1, and TFK-1 cell lines, and may be useful in treating subjects with CCA tumors that have one or more characteristics specific to multiple particular cell lines, e.g., one or more genetic markers, growth rate, and / or cell morphology, e.g., a CCA tumor with characteristics specific to the CC-SW-1 cell line and characteristics specific to the HuCC-TI cell line.

[0032] A feature or combination of features (e.g., a combination of genetic markers, such as mutations) that is specific to a CCA cell line refers to a feature or combination of features that is present in a CCA cell line and is not found in normal (i.e., healthy) cholangiocytes and / or one or more other CCA cell lines.

[0033] In this regard, Example 29 describes the genetic analysis of the above CCA cell lines and the identified mutations in each cell line. Thus, in some embodiments, CCA tumors having one or more characteristics associated with the EGI-1 cell line may have one or more mutations in a gene selected from KRAS, TP53, ASXL1, PDGFRA, MYH11, E2F1, AHNAK, SAFB2, NOTCH1, PEG3, CADM3, SPI1, AR, HCAR2, PPP1R1B, or a combination thereof. In some embodiments, the mutations in these genes are as described in Example 29. In some embodiments, CCA tumors having one or more characteristics associated with the EGI-1 cell line may have one or more mutations in a gene selected from KRAS and / or TP53, particularly KRAS Gly12Asp and / or TP53 Arg273His.

[0034] In some embodiments, CCA 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.

[0035] In some embodiments, CCA tumors having one or more characteristics associated with the HuCC-TI cell line include those expressing KRAS, TP53, FBXW7, LETMD1, SETD2, KDM5A, MYO18B, RB1, DNAJA3, CDT1, ZFP36L2, MAF, GMPS, NPAS2, CNTNAP2, MSH6 (e.g., Lys1358fs), or other mutated CCA tumors expressing IgG4-associated ... *2, coding sequence 4071_4072insGATT), or a combination thereof. In certain embodiments, the mutations in these genes are as described in Example 29. In certain embodiments, CCA tumors having one or more characteristics associated with the HuCC-TI cell line may have one or more mutations in a gene selected from KRAS and / or TP53, particularly KRAS Gly12Asp and / or TP53 Arg175His.

[0036] In some embodiments, CCA tumors having one or more characteristics associated with the CC-SW-1 cell line may have one or more mutations in genes 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.

[0037] As defined herein, "treatment" or "treating" as used herein broadly refers to any effect or step (or intervention) beneficial to the management of a clinical condition or disorder. Thus, treatment can refer to the alleviation, mitigation, improvement, delay in progression, or elimination of one or more symptoms of the cholangiocarcinoma (CCA) being treated compared to the symptoms before treatment, or in any way improving the clinical condition of the subject. Treatment can include clinical steps or interventions that contribute to or are part of a treatment program or regimen. In particular, the treatment can include a reduction in the size or volume of the CCA being treated.

[0038] Treatment can include, for example, delaying, limiting, reducing, or preventing the onset of one or more symptoms of CCA compared to the CCA or symptoms before treatment. Thus, treatment expressly includes both absolute prevention of the onset or development of a CCA symptom and any delay in the onset of CCA or symptoms or reduction or limitation of the onset or progression of CCA or symptoms.

[0039] Thus, treatment according to the present invention includes killing, inhibiting, or slowing the growth of CCA cells, or increasing the size of a body or population of CCA cells (e.g., in a tissue, tumor, or growth), reducing the number of CCA cells or preventing the spread of CCA cells (e.g., to another anatomical site), reducing the size of cell growth, etc. The term "treatment" does not necessarily imply a cure or complete eradication or elimination of CCA cell growth or proliferation of CCA cells.

[0040] A "subject" or "patient" is an animal (ie, any human or non-human animal), preferably a mammal, and most preferably a human.

[0041] The therapeutic agents or drug substances described herein (e.g., 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.

[0042] "Systemic administration" includes any form of non-local administration in which an agent is administered to the body at a site other than immediately adjacent to or locally proximate to the CCA, such that the entire body receives the administered agent. Advantageously, systemic administration can be via enteral delivery (e.g., oral) or parenteral delivery (e.g., intravenous, intramuscular, or subcutaneous).

[0043] 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 together with a pharmacologically (or pharmaceutically) acceptable excipient.

[0044] The additives may include any additive 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 agent, disintegrant, filler, flavor, colorant, glidant, lubricant, preservative, adsorbent and / or sweetener.

[0045] The additives may be selected from, for example, lactic acid, dextrose, sodium metabisulfate, benzyl alcohol, polyethylene glycol, propylene glycol, microcrystalline cellulose, lactose, starch, chitosan, pregelatinized starch, calcium carbonate, calcium sulfate, cellulose, dextrin, dextrin, dextrose, dibasic calcium phosphate dihydrate, tribasic calcium phosphate, gelatin, magnesium carbonate, magnesium oxide, magnesium stearate, maltodextrin, mannitol, powdered cellulose, pregelatinized starch, sodium chloride, sorbitol, propylene glycol, and / or talc. Additives also typically include colorants such as titanium dioxide and various iron oxides.

[0046] The above list of excipients applies to all drug substances described herein (e.g., panobinostat and cytotoxic agents described below) unless otherwise specified.

[0047] The pharmaceutical compositions described herein may be provided in any form known in the art, such as tablets, capsules, coated tablets, liquids, suspensions, tabs, sachets, implants, powders, pellets, emulsions, lyophilized formulations, effervescent formulations, or any mixtures thereof, and may be provided, for example, as gastro-resistant formulations and / or in sustained release forms.

[0048] In a preferred embodiment, the pharmaceutical composition comprising panobinostat, e.g., panobinostat or a salt thereof, is formulated for oral administration, i.e., panobinostat is administered orally to a subject in the methods and uses of the present invention.

[0049] The most preferred dosage form of panobinostat for treating cholangiocarcinoma is in the form of a tablet or capsule. The tablet may be a coated tablet.

[0050] One of the most preferred dosage forms for the treatment of bile duct cancer is in the form of a capsule.

[0051] Panobinostat or its salt can be administered in any suitable dosage range using any suitable dosing regimen. Those skilled in the art will recognize the appropriate dosage range of panobinostat. In some embodiments, panobinostat or its salt is present in a pharmaceutical composition and administered to a subject in its typical dosage range. This can be considered a therapeutically effective amount of panobinostat.

[0052] As described above, panobinostat is used in combination with another therapeutic agent, for example, a cytotoxic agent that enhances the effect of panobinostat. Thus, in some embodiments, panobinostat can be administered at a dose lower than its typical dose range. However, when a low dose of panobinostat is used in combination therapy, it has the same or equivalent therapeutic effect as a high dose of panobinostat. Thus, in some embodiments, the present invention also makes it possible to treat subjects with low or below-average tolerance to panobinostat, such as the elderly, infants, or those who are frail due to disease, malnutrition, etc.

[0053] In a representative embodiment, the clinical dose of panobinostat for the treatment of cholangiocarcinoma is about 5-50 mg, more preferably 10-30 mg, administered daily or at least twice weekly, e.g., 2-6, 2-5, or 2-4 times weekly. In a preferred embodiment, the clinical dose is a single-dose formulation, e.g., a tablet or capsule.

[0054] As described above and in detail in the Examples, the present inventors have found that the effect of panobinostat on CCA may be enhanced by combining it with various other cytotoxic agents, such as anticancer agents.

[0055] Thus, the present invention relates to therapeutic regimens for the treatment of cholangiocarcinoma that combine panobinostat with other cytotoxic agents, such as anti-cancer drugs.

[0056] Thus, an additional cytotoxic agent (e.g., an anticancer agent) described herein can be used to provide a sensitizing effect, i.e., to enhance (or alternatively, increase, augment, or potentiate) the effect of panobinostat (e.g., in treating CCA) or to make a subject (or more specifically, a CCA cell or tumor present in a subject) more susceptible to panobinostat. Thus, in certain embodiments, panobinostat can be considered a first-order drug (therapeutic agent), and the additional cytotoxic agent can be considered a second-order drug (therapeutic agent).

[0057] The terms "primary drug" and "primary therapeutic agent" refer to a drug that is administered at a higher relative dose compared to 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%, e.g., 100%, of the maximum tolerated dose), and the secondary drug is administered at a dose substantially less 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 a dose greater than or equal to the IC 20 Because different agents have different dosage ranges, it will be apparent that the secondary agent may be administered at a higher absolute dose than the primary agent, even if it is administered at a dose substantially less than its maximum tolerated dose.

[0058] The maximum tolerated dose (MTD) refers to the highest dose of a pharmacological treatment that produces a desired effect without unacceptable toxicity. Those skilled in the art will recognize the MTD of any given cytotoxic agent disclosed herein.

[0059] In certain embodiments, the additional cytotoxic agent is an agent that is more potent than the IC of panobinostat alone. 50 Compared with panobinostat's IC 50 It can be any drug that reduces IC 50 can be determined using any suitable method, such as the in vitro methods described in the Examples.

[0060] The following examples demonstrate the IC of some cytotoxic drugs when used in combination with specific doses of panobinostat. 50 Thus, in certain embodiments, panobinostat may enhance the therapeutic effect of an additional cytotoxic agent, e.g., reduce the IC of the additional cytotoxic agent. 50 That is, in certain embodiments, panobinostat can be used (i.e., administered) as a second agent (e.g., at a dose substantially less than its maximum tolerated dose), and the additional cytotoxic agent can be administered as a first agent (e.g., at a dose at or near its maximum tolerated dose). In certain embodiments, the effect of panobinostat on the therapeutic effect of the additional cytotoxic agent, e.g., the IC of the additional cytotoxic agent, can be measured. 50 The reduction in β-blockers may be additive to the effect of additional cytotoxic agents on panobinostat.

[0061] In a preferred embodiment, panobinostat is used as the primary drug in the combinations disclosed herein.

[0062] The term “IC 50 " is a measure of the effectiveness of a substance in inhibiting a particular biological or biochemical function. Thus, in the context of the present invention, IC 50 represents the concentration of a drug (e.g., panobinostat) required for 50% inhibition (reduction) of CCA cell viability in vitro. Similarly, the term "IC 20 " represents the concentration of a drug required for 20% inhibition (reduction) of CCA cell viability in vitro. Therefore, the inhibitory concentration (IC) can be considered the lethal concentration (LC) or lethal dose (LD) of a substance. These terms are used to describe dosage in in vivo studies.

[0063] Cytotoxic agents (i.e., anti-cancer drugs) described herein are generally associated with adverse events in clinical use. The frequency and severity of toxicity and adverse events generally correlate with dose. The higher the dose, the more frequent and severe the side effects. Anti-cancer drugs are generally used at the highest possible clinical dose (maximum tolerated dose) to maximize efficacy. Therefore, when possible, the IC of anti-cancer drugs on cancer cells should be evaluated. 50 Reducing this is clinically important.

[0064] The IC of the primary drug (e.g., panobinostat) in CCA cells when the cytotoxic agent is 50 The ability to reduce the 50 IC of specific cell lines to provide 50 This can be determined by measuring the change in dose. Delta IC 50 This relates to how the monotherapy curve of a particular substance is affected by combination treatment with a second compound. In some examples herein, in various cell lines, second drugs (additional cytotoxic agents) were compared to their IC 20 The secondary drug is added to various concentrations of the primary drug (e.g., panobinostat) at the IC 50 If the secondary agent reduces the IC of panobinostat, the secondary agent can be considered to enhance the effect of the primary agent. As mentioned above, the clinical outcome of the combination is to reduce the dose of the primary agent and reduce the frequency and / or severity of side effects. Another option is to maintain the usual dose of the primary agent to improve the clinical efficacy of the agent for the treatment of CCA. In some embodiments, the additional cytotoxic agent reduces the IC of panobinostat. 50 In certain embodiments, the additional cytotoxic agent may decrease the IC value of panobinostat by at least about 10%, e.g., by at least about 12%, 15%, 20%, 25%, 30%, 40%, or 50%. 50 The value may be reduced by at least about 60%, 70%, 80%, 90% or 100%.

[0065] In certain embodiments, the additional cytotoxic agent is any agent that, when used in combination with panobinostat, is more effective (e.g., additive or synergistic) in treating CCA than panobinostat alone at the same dose or concentration of panobinostat.

[0066] 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 may function synergistically (efficacy is greater than the additive effect 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 is less than the sum of the efficacies of the individual drugs, e.g., 2 + 2 = 3). CI can be calculated using the Chou-Talalay principle using CalcuSyn software (Biosoft, Ferguson, MO) (see also Chou TC, Talalay P. Adv Enzyme Regul. 1984;22:27-55; Lu Huang et al. Nature, Volume 7, Article number: 40752 (2017); and Ashkan Zandi et al. Middle East Journal of Cancer; January 2017;8(1):31-38, all of which are incorporated herein by reference). A CI value less than 1 indicates synergy. A CI value of 1 indicates an additive effect; a CI greater than 1 indicates antagonism. In certain embodiments, the additional cytotoxic agent, if used alone, may be effective in inhibiting (e.g., killing) the viability of CCA cells (e.g., treating CCA in a subject). Thus, in certain embodiments, the effect of the combination of panobinostat and the additional cytotoxic agent on inhibiting the viability of CCA cells (e.g., treating CCA in a subject) is additive, i.e., the combination has a CI of 1.

[0067] Additive interaction means that the effect of panobinostat and the additional cytotoxic agent is equal to the sum of their separate effects at the same dose, where the effect is the ability of the agents to inhibit the viability of (e.g., kill) CCA cells as assessed, for example, using the in vitro assays described in the Examples.

[0068] In certain embodiments, the effect of the combination of panobinostat and an additional cytotoxic agent on inhibiting CCA cell viability (eg, treating CCA in a subject) is synergistic.

[0069] Synergistic interaction means that the effect of panobinostat and the additional cytotoxic agent is greater than the sum of their separate effects at the same dose, e.g., the effect is the ability of the agents to inhibit survival (e.g., kill) of CCA cells, i.e., the combination has a CI of less than 1, e.g., about 0.95, 0.90, 0.85, 0.80, 0.75 or less, as assessed using the in vitro assays described in the Examples.

[0070] In certain embodiments, the combination of panobinostat with an additional cytotoxic agent improves the safety margin of panobinostat for use in the treatment of CCA compared to the use of panobinostat alone for use in the treatment of CCA.

[0071] "Safety ratio" is the ratio between the dose that causes toxic effects and / or serious side effects in a subject and the effective dose (e.g., therapeutically effective amount). Thus, in some embodiments, the safety ratio of the panobinostat combination disclosed herein is higher than the safety ratio when panobinostat alone is used to treat cholangiocarcinoma. Alternatively, in some embodiments, the additional cytotoxic agent is an agent that improves the safety ratio of panobinostat.

[0072] In certain embodiments, the combination of panobinostat with an additional cytotoxic agent improves the therapeutic index of panobinostat for use in the treatment of CCA compared to the use of panobinostat alone for use in the treatment of CCA.

[0073] Therapeutic index (TI) is the toxic dose (TD 50 ) and effective dose (ED 50 ED is a quantitative measure of a drug's relative safety, measured as the ratio of ED to ED. 50 is the dose that produces a therapeutic effect in 50% of patients, and TD 50 is the dose that produces a toxic effect in 50% of patients. These values ​​can be extracted from a dose-response curve. From a clinical point of view, it is advantageous to have as high a therapeutic index as possible. A high therapeutic index value indicates 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), patients will have a much higher probability of having serious side effects using a certain clinical dose. For drugs used in clinical practice, the TI varies from drug to drug. Cytotoxic drugs (e.g., anti-cancer drugs) usually have a low TI, while penicillin and paracetamol have a much higher TI.

[0074] As shown in the examples, TI is the IC between normal cells and cancer cells. 50 Therapeutic index can be calculated using in vitro data based on the ratio between the TI and the TI. Thus, in certain 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 certain embodiments, the in vitro TI of the combination is at least 1.5, preferably 2.0, 2.5, 3.0 or more, or 5, 6, 7, 8, 9, 10 or more.

[0075] The term "drug sensitivity score (DSS)" refers to a quantitative measure for characterizing a drug or combination of drugs in a single parameter. DSS describes a multiparametric dose-response relationship with a single value between 1 and 100, where higher values ​​indicate more effective therapy. DDS identifies selective drug or drug combination responses between cancer cells and control cells (see Yadaveta l. Scientific Reports (Nature) Volume 4, Article number: 5193 (2014)). Thus, in some embodiments, the combination therapy disclosed herein has a higher DSS than a monotherapy, e.g., panobinostat alone.

[0076] "Cytotoxic agent" refers to an agent capable of inhibiting or suppressing the growth, viability, and / or proliferation (replication / proliferation) of animal cells. In certain embodiments, the cytotoxic agent is capable of inhibiting or suppressing (e.g., killing) the growth, viability, and / or proliferation (replication / proliferation) of CCA cells, preferably human CCA cells.

[0077] Included as cytotoxic agents are anti-tumor agents and any agent potentially indicated for oncological use, and thus include agents used in chemotherapy treatment protocols ("chemotherapeutic agents" or "anti-cancer agents").

[0078] Cytotoxic agents are generally classified into different classes according to their mechanism of action, and all of these classes are contemplated herein. Thus, cytotoxic agents can be, for example, alkylating agents, cross-linking agents, intercalating agents, nucleotide analogs, inhibitors of spindle formation, and / or topoisomerase I and / or II inhibitors. Other types or classes of agents include antimetabolites, plant alkaloids and terpenoids, or antitumor antibiotics.

[0079] Alkylating agents modify DNA by alkylating nucleosides, which leads to interference with correct DNA replication. Nucleotide analogs are incorporated into DNA during replication and inhibit DNA synthesis. Spindle formation inhibitors disrupt spindle formation, leading to mitotic arrest during metaphase. Intercalating agents intercalate between DNA bases, thereby inhibiting DNA synthesis. Topoisomerase I or II inhibitors affect DNA twisting, thereby interfering with DNA replication.

[0080] Suitable cytotoxic agents are known in the art, and examples include actinomycin D, bortezomib, BCNU (carmustine), BI2536, buparlisib, carboplatin, CCNU, campothecin (CPT), cantharidin, cisplatin, combretastatin A4, CUDC-907, cyclophosphamide, cytarabine, dasatinib, dacarbazine, dactolisib, dapolinad, daunorubicin, docetaxel, doxorubicin, duvelisib, DTIC, elesclomol, epirubicin, etoposide, gefitinib, gemcitabine, idelalisib, ifosfamide, ispinesib, irinotecan, ionomycin, luminespib, melphalan, Methotrexate, mitomycin C (MMC), mitoxantrone, mercaptopurine, molybreceptor, oxaliplatin, obatoclax, paclitaxel (Taxol), PARP-1 inhibitors, pelitinib, perifosine, PX-866, sepantronium bromide, SB-743921, taselisib, taxotere, temozolomide (TZM), teniposide, topotecan, trametinib, treosulfan, triptolide, umbralisib, vinorelbine, vincristine, vinblastine, volasertib, voxtalisib, 5-azacytidine, 5,6-dihydro-5-azacytidine, and 5-fluorouracil may be used in the combination therapy of the present invention.

[0081] In particularly preferred embodiments, the additional cytotoxic agent is selected from bortezomib, BI2536, carboplatin, cisplatin, combretastatin A4, dactolisib, dapolinad, dasatinib, docetaxel, doxorubicin, elesclomol, gemcitabine, ispinesib, luminespib, methotrexate, molybrespib, obatoclax, pelitinib, SB-743921, topotecan, trametinib, and triptolide.

[0082] In further preferred embodiments, the additional cytotoxic agent is selected from BI2536, carboplatin, cisplatin, combretastatin A4, dactolisib, dapolinad, dasatinib, docetaxel, doxorubicin, elesclomol, ispinesib, luminespib, methotrexate, molybrespib, obatoclax, pelitinib, SB-743921, topotecan, trametinib, and triptolide.

[0083] In other preferred embodiments, the additional cytotoxic agent is selected from doxorubicin, dactolisib, SB-743921, trametinib, elesclomol, molybreceptor, methotrexate, dapolinad, topotecan, cisplatin, dasatinib, carboplatin, and luminespib.

[0084] In yet another preferred embodiment, the additional cytotoxic agent is selected from carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, molybresib, obatoclax, pelitinib, trametinib and triptolide, preferably carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib.

[0085] The cytotoxic agent for use in combination with panobinostat can be provided in the pharmaceutical composition defined above and can be administered as defined above and further below. In some embodiments, the pharmaceutical composition containing the cytotoxic agent can be formulated for parenteral administration. Thus, the composition can include pharmaceutically acceptable additives, solvents and diluents suitable for such formulation, for example, intravenous bolus or infusion.

[0086] Those skilled in the art will know the appropriate dosage range for a given cytotoxic agent. In a preferred embodiment, the cytotoxic agent is present in a pharmaceutical composition or administered to a subject in its typical dosage range.

[0087] However, as shown in the Examples below and as described above, some cytotoxic agents can enhance the effect of panobinostat on CCA cells at low doses. Thus, in some embodiments, an additional cytotoxic agent can be present in a pharmaceutical composition or administered to a subject at a dose range lower than the typical dose ranges described below. For example, in some embodiments, an additional cytotoxic agent can be present in a pharmaceutical composition or administered to a subject at a dose range that is 70% or less of the typical dose range, for example, 60%, 50%, 40%, or 30% or less of the typical dose range (e.g., maximum tolerated dose). Thus, in some embodiments, the therapeutically effective amount of the additional cytotoxic agent is lower than the typical dose range defined above.

[0088] In some embodiments, the combination therapy comprises administration of panobinostat and bortezomib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0089] 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.

[0090] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with bortezomib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0091] In another embodiment, the present invention provides the use of panobinostat, or a pharmaceutically acceptable salt thereof, in the manufacture of a combination product with bortezomib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0092] In certain embodiments, combination therapy of panobinostat and bortezomib is used for the treatment of intrahepatic CCA.

[0093] In certain embodiments, the combination therapy of panobinostat and bortezomib is used to treat a subject having a CCA tumor (e.g., an intrahepatic CCA 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.

[0094] Bortezomib ([(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]propanoyl}amino)butyl]boronic acid) is a proteasome inhibitor having the structure shown below. Bortezomib is available from Janssen. The term "bortezomib" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. [ka]

[0095] Stable liquid pharmaceutical compositions of bortezomib are described in WO2016 / 166653 (hereby incorporated by reference), and any such compositions may be used in the methods, compositions and uses of the present invention.

[0096] In certain embodiments, the composition comprising bortezomib is a "ready to use" formulation containing bortezomib in a dissolved or solubilized form and is intended to be used by itself or upon further dilution with an intravenous diluent.

[0097] In a preferred embodiment, the pharmaceutical composition comprising bortezomib is formulated for parenteral administration, eg, by injection or infusion.

[0098] Suitable solvents may be selected from aqueous and non-aqueous solvents, such as, but not limited to, glycerin, ethanol, n-propanol, n-butanol, isopropanol, ethyl acetate, dimethyl carbonate, acetonitrile, dichloromethane, methyl ethyl ketone, methyl isobutyl ketone, cyclohexane, dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), 1,3-dimethyl-2-imidazolidinone (DMI), acetone, tetrahydrofuran (THF), dimethylformamide (DMF), propylene carbonate (PC), dimethyl isosorbide, water, and mixtures thereof. Preferred solvents are ethanol, glycerin, and water.

[0099] The bortezomib formulations 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.

[0100] The bortezomib formulation 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), TTTA (1,7,13-triaza-4,10,16-trioxacyclooctadecane-N,N',N"-triacetate), DOTRP (tetraethyleneglycol-1,5,9-triazacyclododecane-N,N',N"-tris(methylenephosphonic acid), EGTA (ethyleneglycol-bis(p-aminoethyl ether)-tetraacetic acid), and the like.

[0101] 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 metabisulfite, L-cysteine, thioglycolic acid, citric acid, tartaric acid, phosphoric acid, gluconic acid, and thiodipropionic acid. The most preferred antioxidant is monothioglycerol.

[0102] The most preferred mode of administration of the combination of panobinostat and bortezomib for the treatment of cholangiocarcinoma is bortezomib in the form of subcutaneous or intravenous injection.

[0103] The bortezomib injection used in the present invention is preferably in the form of a water-soluble boronic acid ester, the most preferred ester being mannitol boronic acid ester.

[0104] The boronate ester formulations, preferably mannitol esters, are typically in the form of sterile dry powder formulations. The powders are typically lyophilized powders that are dissolved in sterile water, typically sterile isotonic aqueous sodium chloride solution, prior to administration.

[0105] The bortezomib formulations used in the present invention may optionally contain other pharmaceutically acceptable adjuvants such as buffering agents, pH adjusting agents, preservatives, tonicity modifiers, and the like.

[0106] The above list of solvents, stabilizers, chelating agents and antioxidants may also be used in pharmaceutical compositions containing other cytotoxic agents described herein, unless otherwise specified.

[0107] The bortezomib-based formulation preferably contains mannitol and is provided in injection vials or pre-filled syringes under a nitrogen atmosphere.

[0108] A preferred embodiment of the use of a combination of panobinostat and bortezomib for the treatment of cholangiocarcinoma is to administer panobinostat orally and bortezomib in the form of an injection.

[0109] In certain embodiments, the clinical dose of panobinostat in combination with bortezomib for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0110] In certain embodiments, the clinical dose of bortezomib in combination with panobinostat for the treatment of cholangiocarcinoma is generally 0.5 to 3 mg / m 2 Body surface area (BSA) at least once a week, preferably 1-2 mg / m 2 Body surface area (BSA) is measured at least once a week.

[0111] A preferred embodiment of the present invention in which a combination of panobinostat and bortezomib is administered for the treatment of cholangiocarcinoma is with the co-administration of a glucocorticosteroid, typically dexamethasone.

[0112] In some embodiments, the combination therapy comprises administration of panobinostat and carboplatin. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0113] 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.

[0114] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with carboplatin, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in a subject.

[0115] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with carboplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0116] In certain embodiments, the combination therapy of panobinostat and carboplatin is used for the treatment of extrahepatic CCA.

[0117] In certain embodiments, the combination therapy of panobinostat and carboplatin is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably the CC-SW-1 cell line and / or TFK-1 cell line.

[0118] Carboplatin (cis-(1,1-cyclobutanedicarboxylato)diammineplatinum(II)) is a platinum-containing anticancer drug having the structure shown below. Carboplatin is widely available. The term "carboplatin" includes its pharmaceutically acceptable salts, solvates, and hydrates. [ka]

[0119] 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.

[0120] In certain embodiments, the carboplatin-containing composition is a "ready-to-use" formulation containing carboplatin in a dissolved or solubilized form, intended for use by itself or after further dilution with an intravenous diluent.

[0121] In a preferred embodiment, the pharmaceutical formulation containing carboplatin is intended for parenteral administration.

[0122] A preferred embodiment of the use of a combination of panobinostat and carboplatin for the treatment of cholangiocarcinoma is to administer panobinostat orally and carboplatin in the form of an injection or infusion.

[0123] In certain embodiments, the clinical dose of panobinostat in combination with carboplatin for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0124] In certain embodiments, the clinical dose of carboplatin in combination with panobinostat for the treatment of cholangiocarcinoma will generally be in the same range as that currently used when carboplatin is used for other indications, e.g., 1 to 30 mg / m 2 BSA. The Calvert formula should be used to calculate the exact clinical dose.

[0125] In some embodiments, the combination therapy comprises administration of panobinostat and cisplatin. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0126] 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.

[0127] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with cisplatin, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in a subject.

[0128] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with cisplatin or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0129] In certain embodiments, the combination therapy of panobinostat and cisplatin is used for the treatment of extrahepatic CCA.

[0130] In certain embodiments, the combination therapy of panobinostat and cisplatin is used to treat a subject having a CCA tumor having one or more characteristics specific to the CC-SW-1 and / or TFK-1 cell lines, e.g., one or more genetic markers, growth rate, and / or cell morphology.

[0131] Cisplatin ((SP-4-2)-diamminedichloroplatinum(II)) is a platinum-containing anticancer drug having the structure shown below. Cisplatin is widely available from companies such as Hospira. The term "cisplatin" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. [ka]

[0132] 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.

[0133] In certain embodiments, the composition comprising cisplatin is a "ready-to-use" formulation containing cisplatin in a dissolved or solubilized form, and is intended to be used by itself or after further dilution with an intravenous diluent.

[0134] In a preferred embodiment, the pharmaceutical formulation containing cisplatin is intended for parenteral administration.

[0135] A preferred embodiment of the use of a combination of panobinostat and cisplatin for the treatment of cholangiocarcinoma is to administer panobinostat orally and cisplatin in the form of an injection or infusion.

[0136] In certain embodiments, the clinical dose of panobinostat in combination with cisplatin for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0137] In certain embodiments, the clinical dose of cisplatin in combination with panobinostat for the treatment of cholangiocarcinoma will generally be in the same range as that currently used when cisplatin is used for other indications, e.g., 10-50 mg / m 2 BSA, preferably 20-30 mg / m 2BSA. The Calvert formula should be used to calculate the exact clinical dose.

[0138] In some embodiments, the combination therapy comprises administration of panobinostat and dasatinib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0139] 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.

[0140] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with dasatinib, or a pharmaceutically acceptable salt thereof, for separate, simultaneous or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0141] In another embodiment, the present invention provides the use of panobinostat, or a pharmaceutically acceptable salt thereof, in the manufacture of a combination product with dasatinib, or a pharmaceutically acceptable salt thereof, for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0142] In certain embodiments, the panobinostat and dasatinib combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and dasatinib in a single dosage form (eg, a tablet or capsule).

[0143] In certain embodiments, combination therapy of panobinostat and dasatinib is used for the treatment of extrahepatic CCA.

[0144] In certain embodiments, the combination therapy of panobinostat and dasatinib is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably the CC-SW-1 cell line and / or TFK-1 cell line.

[0145] 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 and is disclosed in WO2000 / 062778 (Formula I). ​​Dasatinib has the structure shown below. Dasatinib is available from Bristol-Myers Squibb. The term "dasatinib" includes pharmaceutically acceptable salts and hydrates thereof. [ka]

[0146] Pharmaceutical compositions of dasatinib are well known in the literature, for example WO2000 / 062778, WO2007 / 035874 and WO2015 / 181573 (all incorporated herein by reference), and any such compositions may be used in the methods, compositions and uses of the present invention.

[0147] In a preferred embodiment, a pharmaceutical composition comprising dasatinib is formulated for oral administration.

[0148] A preferred embodiment of the combination of panobinostat and dasatinib in combination with panobinostat for the treatment of cholangiocarcinoma is to administer both panobinostat and dasatinib orally.

[0149] Thus, in some embodiments, panobinostat and dasatinib can be administered in separate dosage forms (e.g., separate tablets or capsules), hi some embodiments, panobinostat and dasatinib can be administered in a single dosage form (e.g., tablet or capsule) as a combined formulation.

[0150] A formulation (pharmaceutical composition as defined herein) containing both panobinostat and dasatinib in the same combination (eg, tablet or capsule) for the treatment of cholangiocarcinoma is an embodiment of the present invention.

[0151] The most preferred mode of administration of the combination of panobinostat and dasatinib for the treatment of cholangiocarcinoma is dasatinib in the form of an oral formulation containing dasatinib monohydrate.

[0152] Exemplary oral formulations of dasatinib for treating cholangiocarcinoma according to the present invention include at least one of the following excipients: lactose, mannitol, microcrystalline cellulose, hydroxypropyl methylcellulose (HPMC), cross-linked sodium carboxymethylcellulose, magnesium stearate, sodium lauryl sulfate, polyethylene glycol, and silicon dioxide.

[0153] In certain embodiments, the clinical dose of panobinostat in combination with dasatinib for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0154] In certain embodiments, the clinical dose of dasatinib for the treatment of cholangiocarcinoma is generally 10-200 mg / day.

[0155] In some embodiments, the combination therapy comprises administration of panobinostat and doxorubicin. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0156] 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.

[0157] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with doxorubicin, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in a subject.

[0158] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with doxorubicin or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0159] The combination therapy of panobinostat and doxorubicin can be used to treat intrahepatic or extrahepatic CCA. In some embodiments, the combination therapy of panobinostat and doxorubicin is used to treat intrahepatic CCA.

[0160] In certain embodiments, combination therapy of panobinostat and doxorubicin is used to treat a subject having a CCA tumor with one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1, HuCC-T1, EFI-1, and / or TFK-1 cell lines.

[0161] Doxorubicin ((1S,3S)-3-glycolyl-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-lyxo-hexopyranoside) is a cytotoxic antibiotic drug substance having the structure shown below. Doxorubicin is widely available from companies such as Janssen and Pfizer. The term "doxorubicin" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. [ka]

[0162] 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 present invention.

[0163] In certain embodiments, the compositions comprising doxorubicin are "ready-to-use" formulations containing doxorubicin in a dissolved or solubilized form and are intended for use by itself or upon further dilution with an intravenous diluent.

[0164] In a preferred embodiment, the pharmaceutical composition comprising doxorubicin is formulated for parenteral administration.

[0165] A preferred embodiment of the use of a combination of panobinostat and doxorubicin for the treatment of cholangiocarcinoma is to administer panobinostat orally and doxorubicin in the form of an injection or infusion.

[0166] In certain embodiments, the clinical dose of panobinostat in combination with doxorubicin for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0167] In certain embodiments, the clinical dose of doxorubicin in combination with panobinostat for the treatment of cholangiocarcinoma will generally be in the same range as that currently used when using doxorubicin for other indications, e.g., 10-100 mg / m 2 Body surface area (BSA), preferably 40-75 mg / m 2 BSA is administered every 2 to 4 weeks.

[0168] In some embodiments, the combination therapy comprises administration of panobinostat and gemcitabine. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0169] 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.

[0170] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with gemcitabine, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0171] In another embodiment, the present invention provides the use of panobinostat, or a pharmaceutically acceptable salt thereof, in the manufacture of a combination product with gemcitabine, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0172] The combination therapy of panobinostat and gemcitabine can be used to treat intrahepatic or extrahepatic CCA. In some embodiments, the combination therapy of panobinostat and gemcitabine is used to treat intrahepatic CCA.

[0173] In certain embodiments, the combination therapy of panobinostat and gemcitabine is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably the CC-SW-1 cell line and / or HuCC-T1 cell line.

[0174] 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, such as from Eli Lilly & Co (Gemzar®) or Sigma-Aldrich, St. Louis, MO, USA. The term "gemcitabine" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above, preferably the hydrochloride salt. [ka]

[0175] 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.

[0176] In certain embodiments, the gemcitabine-containing composition is a "ready-to-use" formulation containing gemcitabine in a dissolved or solubilized form and is intended for use by itself or upon further dilution with an intravenous diluent.

[0177] In a preferred embodiment, the pharmaceutical composition comprising gemcitabine is formulated for parenteral administration.

[0178] A preferred embodiment of the use of a combination of panobinostat and gemcitabine for the treatment of bile duct cancer is to administer panobinostat orally and gemcitabine in the form of an injection or infusion.

[0179] In certain embodiments, the clinical dose of panobinostat in combination with gemcitabine for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0180] In certain embodiments, the clinical dose of gemcitabine in combination with panobinostat for the treatment of cholangiocarcinoma will generally be in the same range as that currently used when gemcitabine is used for other indications, e.g., 500 to 1500 mg / m 2 (This is the body surface area, BSAm 2 The preferred dose is 900-1100 mg / m 2 Conveniently, gemcitabine may be administered over a period of less than 1 hour, for example 15 to 45 minutes, for example about 30 minutes, or over a longer time frame, for example 1 to 12 hours.

[0181] In some embodiments, the combination therapy comprises administration of panobinostat and methotrexate. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0182] 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.

[0183] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with methotrexate, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in a subject.

[0184] In another embodiment, the present invention provides the use of panobinostat, or a pharmaceutically acceptable salt thereof, in the manufacture of a combination product with methotrexate, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0185] In certain embodiments, the panobinostat and methotrexate combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and methotrexate in a single dosage form (eg, a tablet or capsule).

[0186] In certain embodiments, combination therapy of panobinostat and methotrexate is used for the treatment of extrahepatic CCA.

[0187] In certain embodiments, combination therapy of panobinostat and methotrexate is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, and / or TFK-1 cell line, preferably the TFK-1 cell line.

[0188] 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, such as from Hospira, Inc. The term "methotrexate" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above, and are preferably sodium salts. [ka]

[0189] 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.

[0190] In certain embodiments, the methotrexate-containing composition is a "ready-to-use" formulation containing methotrexate in a dissolved or solubilized form and is intended for use by itself or upon further dilution with an intravenous diluent.

[0191] Thus, in certain embodiments, pharmaceutical compositions comprising methotrexate are formulated for parenteral administration, e.g., by injection or infusion. In these embodiments, methotrexate may be provided in the form of a salt, preferably the sodium salt.

[0192] However, in certain embodiments, the pharmaceutical composition comprising methotrexate is formulated for oral administration, eg, as a tablet or capsule.

[0193] In one embodiment, the use of a combination of panobinostat and methotrexate for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and methotrexate is administered in the form of an injection or infusion.

[0194] In another embodiment, the use of a combination of panobinostat and methotrexate for the treatment of cholangiocarcinoma is one in which both panobinostat and methotrexate are administered orally.

[0195] Thus, in some embodiments, panobinostat and methotrexate may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and methotrexate may be administered in a single dosage form (e.g., tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0196] Thus, formulations (pharmaceutical compositions) comprising both panobinostat and methotrexate for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0197] In certain embodiments, the clinical dose of panobinostat in combination with methotrexate for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0198] In certain embodiments, the clinical dose of methotrexate in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as methotrexate currently used for other indications. For example, in certain embodiments, the dose range of methotrexate is 2.5 to 50 mg / m 2 BSA, e.g., 7.5–25 mg / m 2 BSA can be administered weekly.

[0199] In certain embodiments, the combination therapy comprises administration of panobinostat and topotecan. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0200] 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.

[0201] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with topotecan, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0202] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with topotecan or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0203] In certain embodiments, the panobinostat and topotecan combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and topotecan in a single dosage form (eg, a tablet or capsule).

[0204] Combination therapy of panobinostat and topotecan can be used to treat intrahepatic or extrahepatic CCA.

[0205] In certain embodiments, the combination therapy of panobinostat and topotecan is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably the CC-SW-1 cell line and / or TFK-1 cell line.

[0206] Topotecan (9-[(dimethylamino)methyl]-10-hydroxy-(4S)-camptothecin) is a topoisomerase inhibitor having the structure shown below. Topotecan is widely available, such as from Actavis. The term "topotecan" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above, preferably the hydrochloride salt. [ka]

[0207] 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.

[0208] In certain embodiments, the composition comprising topotecan is a "ready-to-use" formulation containing topotecan in a dissolved or solubilized form, intended for use by itself or after further dilution with an intravenous diluent.

[0209] Thus, in certain embodiments, a pharmaceutical composition comprising topotecan is formulated for parenteral administration, eg, by injection or infusion.

[0210] However, in certain embodiments, a pharmaceutical composition comprising topotecan is formulated for oral administration, eg, as a tablet or capsule.

[0211] In one embodiment, the use of a combination of panobinostat and topotecan for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and topotecan is administered in the form of an injection or infusion.

[0212] In another embodiment, the use of a combination of panobinostat and topotecan for the treatment of cholangiocarcinoma is one in which both panobinostat and topotecan are administered orally.

[0213] Thus, in some embodiments, panobinostat and topotecan may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and topotecan may be administered in a single dosage form (e.g., tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0214] Thus, formulations (ie pharmaceutical compositions) containing both panobinostat and topotecan in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0215] In certain embodiments, the clinical dose of panobinostat in combination with topotecan for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0216] In certain embodiments, the clinical dose of topotecan in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for topotecan for other indications. For example, in certain embodiments, the dose range for topotecan is 0.25 to 3 mg / m, as defined above. 2 BSA, e.g., 0.75-1.50 mg / m 2 BSA can be administered daily or at least twice a week.

[0217] In some embodiments, the combination therapy comprises administration of panobinostat and trametinib.Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0218] 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.

[0219] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with trametinib, or a pharmaceutically acceptable salt thereof, for separate, simultaneous or sequential use or administration to a subject for use in treating cholangiocarcinoma in a subject.

[0220] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with trametinib or a pharmaceutically acceptable salt thereof for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0221] In certain embodiments, the panobinostat and trametinib combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and trametinib in a single dosage form (eg, a tablet or capsule).

[0222] Combination therapy of panobinostat and trametinib can be used to treat extrahepatic or intrahepatic CCA.

[0223] In certain embodiments, the combination therapy of panobinostat and trametinib is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line and / or TFK-1 cell line, preferably the CC-SW-1 cell line and / or TFK-1 cell line.

[0224] Trametinib is a tyrosine kinase inhibitor with affinity for mitogen-activated protein kinases, having the structure shown below. Trametinib is available from Novartis. The term "trametinib," as defined elsewhere herein, includes pharmaceutically acceptable salts thereof. In some embodiments, trametinib is provided in the form of trametinib dimethyl sulfoxide. [ka]

[0225] 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 present invention.

[0226] In a preferred embodiment, a pharmaceutical composition comprising trametinib is formulated for oral administration (eg, a tablet or capsule).

[0227] A preferred embodiment of the combination of panobinostat and trametinib for the treatment of cholangiocarcinoma is to administer both panobinostat and trametinib orally.

[0228] Thus, 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 in a single dosage form (e.g., tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0229] Formulations (ie pharmaceutical compositions) comprising both panobinostat and trametinib in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the invention.

[0230] Certain preferred embodiments of the invention, in which a combination of panobinostat and trametinib is administered for the treatment of cholangiocarcinoma, relate to the use of an oral trametinib formulation, wherein trametinib is optionally in the form of dimethyl sulfate solvate, and the oral formulation contains one or more of the following excipients: mannitol, microcrystalline cellulose, hydroxypropyl methylcellulose (HPMC), cross-linked sodium carboxymethylcellulose, magnesium stearate, sodium lauryl sulfate, and silicon dioxide.

[0231] In certain embodiments, the clinical dose of panobinostat in combination with trametinib for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0232] In some embodiments, the clinical dose of trametinib in combination with panobinostat for the treatment of cholangiocarcinoma is generally 0.1 to 10 mg, more preferably 0.5 to 5 mg, administered daily or at least twice a week, for example, 2 to 6 times, 2 to 5 times, or 2 to 4 times. In a preferred embodiment, the clinical dose is a single-dose formulation, such as a tablet or capsule.

[0233] In some embodiments, the combination therapy comprises administration of panobinostat and combretastatin A4. Thus, the present invention provides a method for treating cholangiocarcinoma in a subject, 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 combretastatin A4 or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0234] Combretastatin A4 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.

[0235] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with combretastatin A4, or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0236] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with combretastatin A4 or a pharmaceutically acceptable salt, solvate or hydrate thereof for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0237] In certain embodiments, the panobinostat and combretastatin A4 combination product is a fixed dose, eg, a pharmaceutical composition, comprising panobinostat and combretastatin A4 in a single dosage form (eg, a tablet or capsule).

[0238] In certain embodiments, combination therapy of panobinostat and combretastatin A4 may be used to treat intrahepatic CCA.

[0239] In one embodiment, combination therapy of panobinostat and combretastatin A4 is used to treat a subject having a CCA tumor with 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.

[0240] Combretastatin A4 (2-methoxy-5-[(Z)-2-(3,4,5-trimethoxy-phenyl)-vinyl]-phenol) is a stilbenoid having the structure shown below. It can be isolated from African bush willow (Combretum caffrum). The term "combretastatin A4" includes its pharmaceutically acceptable salts, solvates, and hydrates. The pharmaceutically acceptable salts, solvates, and hydrates are preferably as defined herein above. In some embodiments, combretastatin A4 is provided in the form of a water-soluble ester, such as a water-soluble phosphate ester. [ka]

[0241] Combretastatin A4 may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0242] In certain embodiments, the composition comprising combretastatin A4 is a "ready-to-use" formulation containing combretastatin A4 in a dissolved or solubilized form and is intended to be used by itself or after further dilution with an intravenous diluent.

[0243] Thus, in certain embodiments, a pharmaceutical composition comprising combretastatin A4 is formulated for parenteral administration, eg, by injection or infusion.

[0244] However, in certain embodiments, a pharmaceutical composition comprising combretastatin A4 is formulated for oral administration, eg, as a tablet or capsule.

[0245] In one embodiment, the use of a combination of panobinostat and combretastatin A4 for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and combretastatin A4 is administered in the form of injection or infusion.

[0246] In another embodiment, the use of a combination of panobinostat and combretastatin A4 for the treatment of cholangiocarcinoma is one in which both panobinostat and combretastatin A4 are administered orally.

[0247] Thus, in some embodiments, panobinostat and combretastatin A4 may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and combretastatin A4 may be administered in a single dosage form (e.g., a tablet or capsule) as a combined formulation (i.e., a pharmaceutical composition).

[0248] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and combretastatin A4 in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0249] In certain embodiments, the clinical dose of panobinostat in combination with combretastatin A4 for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0250] In certain embodiments, the clinical dose of combretastatin A4 in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for combretastatin A4 for other indications. For example, in certain embodiments, the dose range of combretastatin A4 is 5 to 100 mg / m, as defined above. 2 BSA, e.g., 20–85 mg / m 2 BSA can be administered daily or at least twice weekly.

[0251] In certain embodiments, the combination therapy comprises administration of panobinostat and SB-743921. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0252] SB-743921, 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.

[0253] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with SB-743921, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0254] In another embodiment, the present invention provides the use of panobinostat, or a pharmaceutically acceptable salt thereof, in the manufacture of a combination product with SB-743921, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0255] The combination therapy of panobinostat and SB-743921 can be used to treat intrahepatic or extrahepatic CCA. In some embodiments, the combination therapy of panobinostat and SB-743921 is used to treat extrahepatic CCA.

[0256] In certain embodiments, the combination therapy of panobinostat and SB-743921 is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably the CC-SW-1 cell line and / or EGI-1 cell line.

[0257] SB-743921 is a cell division kinesin KSP inhibitor having the structure shown below. The code name "SB-743921" includes its pharmaceutically acceptable salts, solvates, and hydrates. The pharmaceutically acceptable salts are preferably as defined above, preferably the hydrochloride salt. [ka]

[0258] 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.

[0259] In certain embodiments, the composition comprising SB-743921 is a "ready-to-use" formulation containing SB-743921 in a dissolved or solubilized form and is intended to be used by itself or with further dilution with an intravenous diluent.

[0260] In a preferred embodiment, a pharmaceutical composition comprising SB-743921 is formulated for parenteral administration.

[0261] A preferred embodiment of the use of a combination of panobinostat and SB-743921 for the treatment of cholangiocarcinoma is to administer panobinostat orally and SB-743921 in the form of an injection or infusion.

[0262] In certain embodiments, the clinical dose of panobinostat in combination with SB-743921 for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0263] In certain embodiments, the clinical dose of SB-743921 in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for SB-743921 for other indications. For example, in certain embodiments, the dose range for SB-743921 is 1 to 10 mg / m 2 BSA can be administered weekly or monthly, for example, every 1-4 weeks.

[0264] In some embodiments, the combination therapy comprises administration of panobinostat and dapolinad. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 dapolinad or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0265] Dapolinad 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] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with dapolinad, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

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

[0268] In some embodiments, the panobinostat and dapolinad combination product is a combination drug, eg, a pharmaceutical composition, comprising panobinostat and dapolinad in a single dosage form (eg, a tablet or capsule).

[0269] The combination therapy of panobinostat and dapolinad can be used to treat intrahepatic or extrahepatic CCA. In some embodiments, the combination therapy of panobinostat and dapolinad is used to treat intrahepatic CCA.

[0270] In certain embodiments, the combination therapy of panobinostat and dapolinad is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1, EGI-1, and / or TFK-1 cell lines.

[0271] Dapolinad ((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 "dapolinad" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above, preferably the hydrochloride salt. [ka]

[0272] Dapolinad may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0273] In some embodiments, the composition containing dapolinad is a "ready-to-use" formulation containing dapolinad in dissolved or solubilized form and is intended for use by itself or after further dilution with an intravenous diluent.

[0274] Thus, in some embodiments, a pharmaceutical composition comprising dapolinad is formulated for parenteral administration, eg, by injection or infusion.

[0275] However, in certain embodiments, a pharmaceutical composition comprising dapolinad is formulated for oral administration, such as a tablet or capsule.

[0276] In one embodiment, the use of a combination of panobinostat and dapolinad for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and dapolinad is administered in the form of an injection or infusion.

[0277] In another embodiment, the use of a combination of panobinostat and dapolinad for the treatment of cholangiocarcinoma is one in which both panobinostat and dapolinad are administered orally.

[0278] Thus, in some embodiments, panobinostat and dapolinad may be administered in separate dosage forms (e.g., separate tablets or capsules), or in some embodiments, panobinostat and dapolinad may be administered in a single dosage form (e.g., tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0279] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and dapolinad in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0280] In certain embodiments, the clinical dose of panobinostat in combination with dapolinad for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, administered daily or at least twice a week, as defined above.

[0281] In some embodiments, the clinical dose of dapolinad in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for dapolinad for other indications. For example, in some embodiments, the dose range for dapolinad is 0.1 to 10 mg / m 2 The BSA can be administered weekly or monthly, for example every 1-6 weeks, 1-5 weeks, 1-4 weeks or 1-3 weeks.

[0282] In some embodiments, the combination therapy comprises administration of panobinostat and ispinesib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 ispinesib or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0283] Ispinesib 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.

[0284] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with ispinesib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0285] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with ispinesib or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0286] In certain embodiments, the panobinostat and ispinesib combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and ispinesib in a single dosage form (eg, a tablet or capsule).

[0287] In certain embodiments, combination therapy of panobinostat and ispinesib may be used to treat extrahepatic CCA.

[0288] In certain embodiments, the combination therapy of panobinostat and ispinesib is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1, EGI-1, and / or TFK-1 cell lines, preferably the CC-SW-1 and / or TFK-1 cell lines.

[0289] Ispinesib (N-(3-aminopropyl)-N-[(1R)-1-[7-chloro-4-oxo-3-(phenylmethyl)-2-quinazolinyl]-2-methylpropyl]-4-methylbenzamide) is a quinazolinone derivative that selectively inhibits the mitotic motor protein, kinesin spindle protein (KSP). Ispinesib has the structure shown below. The term "ispinesib" includes its pharmaceutically acceptable salts, solvates, and hydrates. For example, in some embodiments, ispinesib may be in the form of a hydrochloride salt. In some preferred embodiments, ispinesib is in the form of a free compound. [ka]

[0290] Ispinesib may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0291] In certain embodiments, the composition comprising ispinesib is a "ready-to-use" formulation containing ispinesib in a dissolved or solubilized form and is intended for use by itself or upon further dilution with an intravenous diluent.

[0292] Thus, in certain embodiments, a pharmaceutical composition comprising ispinesib is formulated for parenteral administration, e.g., by injection or infusion.

[0293] However, in certain embodiments, a pharmaceutical composition comprising ispinesib is formulated for oral administration, e.g., as a tablet or capsule.

[0294] In one embodiment, the use of a combination of panobinostat and ispinesib for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and ispinesib is administered in the form of an injection or infusion.

[0295] In another embodiment, the use of a combination of panobinostat and ispinesib for the treatment of cholangiocarcinoma is where both panobinostat and ispinesib are administered orally.

[0296] Thus, in some embodiments, panobinostat and ispinesib may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and ispinesib may be administered in a single dosage form (e.g., a tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0297] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and ispinesib in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0298] In certain embodiments, the clinical dose of panobinostat in combination with ispinesib for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0299] In certain embodiments, the clinical dose of ispinesib in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as ispinesib currently used for other indications. For example, in certain embodiments, the dose range of ispinesib is 5 to 30 mg / m 2 The BSA can be administered weekly or monthly, for example every 1-6 weeks, 1-5 weeks, 1-4 weeks or 1-3 weeks.

[0300] In some embodiments, the combination therapy comprises administration of panobinostat and luminespib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 luminespib or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0301] Luminespib 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.

[0302] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with luminespib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0303] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with luminespib or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0304] In certain embodiments, the panobinostat and luminespib combination product is a fixed dose, eg, a pharmaceutical composition, that includes panobinostat and luminespib in a single dosage form (eg, a tablet or capsule).

[0305] In certain embodiments, combination therapy of panobinostat and luminespib may be used to treat extrahepatic CCA.

[0306] In one embodiment, the combination therapy of panobinostat and luminespib is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate and / or cell morphology, specific for the EGI-1 cell line and / or the TFK-1 cell line, preferably the EGI-1 cell line.

[0307] Luminespib (5-(2,4-dihydroxy-5-isopropyl-phenyl)-N-ethyl-4-[4-(morpholinomethyl)phenyl]isoxazole-3-carboxamide) is an HSP90 inhibitor having the structure shown below. The term "luminespib" includes its pharmaceutically acceptable salts, solvates, and hydrates. Pharmaceutically acceptable salts are preferably as defined above. For example, in some embodiments, luminespib can be in the form of a hydrochloride or methanesulfonate salt. [ka]

[0308] Luminespib may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0309] In some embodiments, the composition comprising luminespib is a "ready-to-use" formulation containing luminespib in dissolved or solubilized form and is intended to be used by itself or after further dilution with an intravenous diluent.

[0310] Thus, in some embodiments, a pharmaceutical composition comprising luminespib is formulated for parenteral administration, eg, by injection or infusion.

[0311] However, in certain embodiments, a pharmaceutical composition comprising luminespib is formulated for oral administration, e.g., as a tablet or capsule.

[0312] In one embodiment, the use of a combination of panobinostat and luminespib for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and luminespib is administered in the form of an injection or infusion.

[0313] In another embodiment, the use of a combination of panobinostat and luminespib for the treatment of cholangiocarcinoma is one in which both panobinostat and luminespib are administered orally.

[0314] Thus, in some embodiments, panobinostat and luminespib may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and luminespib may be administered in a single dosage form (e.g., a tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0315] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and luminespib in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0316] In certain embodiments, the clinical dose of panobinostat in combination with luminespib for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0317] In certain embodiments, the clinical dose of luminespib in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for luminespib for other indications. For example, in certain embodiments, the dose range for luminespib is 5 to 150 mg / m 2 BSA, e.g., 40–70 mg / m 2 The BSA can be administered weekly, for example, every 1-4 weeks, 1-3 weeks, or 1-2 weeks.

[0318] In some embodiments, the combination therapy comprises administration of panobinostat and molybresib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 molybresib or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0319] Molybresive 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.

[0320] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with molybresive, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0321] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with molybresive or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0322] In certain embodiments, the panobinostat and molybresib combination product is a fixed dose, eg, a pharmaceutical composition, that includes panobinostat and molybresib in a single dosage form (eg, a tablet or capsule).

[0323] In certain embodiments, combination therapy of panobinostat and molybresib may be used to treat extrahepatic CCA.

[0324] In one embodiment, the combination therapy of panobinostat and molybresib is used to treat a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the EGI-1 cell line and / or the TFK-1 cell line, preferably the TFK-1 cell line.

[0325] Molybresive (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 extraterminal) family of bromodomain-containing proteins, having the structure shown below. The term "molybresive" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above. For example, in some embodiments, molybresive can be in the form of a hydrochloride or methanesulfonate salt. [ka]

[0326] Molybresib may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0327] In some embodiments, the composition comprising molybresive is a "ready-to-use" formulation containing molybresive in a dissolved or solubilized form and is intended for use by itself or with further dilution with an intravenous diluent.

[0328] Thus, in some embodiments, a pharmaceutical composition comprising molybresib is formulated for parenteral administration, e.g., by injection or infusion.

[0329] However, in certain embodiments, a pharmaceutical composition comprising molybresib is formulated for oral administration, e.g., as a tablet or capsule.

[0330] In one embodiment, the use of a combination of panobinostat and molybresib for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and molybresib is administered in the form of an injection or infusion.

[0331] In another embodiment, the use of a combination of panobinostat and molybresib for the treatment of cholangiocarcinoma is one in which both panobinostat and molybresib are administered orally.

[0332] Thus, in some embodiments, panobinostat and molybresib may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and molybresib may be administered in a single dosage form (e.g., a tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0333] Thus, formulations (ie pharmaceutical compositions) containing both panobinostat and molybresib in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0334] In certain embodiments, the clinical dose of panobinostat in combination with molybreceptor for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0335] In some embodiments, the clinical dose of molybresil in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for other indications. For example, in some embodiments, the dose range of molybresil can be 5 to 150 mg, for example, 10 to 80 mg daily.

[0336] In certain embodiments, the combination therapy comprises administration of panobinostat and pelitinib.Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0337] Pelitinib, 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.

[0338] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with pelitinib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0339] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with pelitinib or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0340] In certain embodiments, the panobinostat and pelitinib combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and pelitinib in a single dosage form (eg, a tablet or capsule).

[0341] In certain embodiments, combination therapy of panobinostat and pelitinib may be used to treat extrahepatic CCA.

[0342] In certain embodiments, the combination therapy of panobinostat and trametinib is used to treat a subject having a CCA tumor with one or more characteristics specific to the TFK-1 cell line, e.g., one or more genetic markers, growth rate, and / or cell morphology.

[0343] Pelitinib ((2E)-N-(4-((3-chloro-4-fluorophenyl)amino)-3-cyano-7-ethoxy-6-quinolinyl)-4-(dimethylamino)-2-butenamide) is an irreversible inhibitor of epidermal growth factor receptor (EGFR) having the structure shown below. The term "pelitinib" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above. For example, in certain embodiments, pelitinib can be in the form of an acid salt, such as a hydrochloride or methanesulfonate salt. [ka]

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

[0345] In certain embodiments, the composition comprising pelitinib is a "ready-to-use" formulation containing pelitinib in a dissolved or solubilized form and is intended to be used by itself or upon further dilution with an intravenous diluent.

[0346] Thus, in certain embodiments, a pharmaceutical composition comprising pelitinib is formulated for parenteral administration, e.g., by injection or infusion.

[0347] However, in certain embodiments, a pharmaceutical composition comprising pelitinib is formulated for oral administration, e.g., as a tablet or capsule.

[0348] In one embodiment, the use of a combination of panobinostat and pelitinib for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and pelitinib is administered in the form of an injection or infusion.

[0349] In another embodiment, the use of a combination of panobinostat and pelitinib for the treatment of cholangiocarcinoma is one in which both panobinostat and pelitinib are administered orally.

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

[0351] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and pelitinib in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0352] In certain embodiments, the clinical dose of panobinostat in combination with pelitinib for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0353] In certain embodiments, the clinical dose of pelitinib in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for pelitinib for other indications. For example, in certain embodiments, the dose range of pelitinib can be 10 to 100 mg, e.g., 25 to 75 mg daily.

[0354] In some embodiments, the combination therapy comprises administration of panobinostat and triptolide. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0355] 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.

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

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

[0358] In certain embodiments, the panobinostat and triptolide combination product is a combination drug, eg, a pharmaceutical composition, that includes panobinostat and triptolide in a single dosage form (eg, a tablet or capsule).

[0359] In certain embodiments, combination therapy of panobinostat and triptolide may be used to treat extrahepatic CCA.

[0360] In certain embodiments, combination therapy of panobinostat and triptolide is used to treat a subject having a CCA tumor with one or more characteristics specific to the TFK-1 cell line, e.g., one or more genetic markers, growth rate, and / or cell morphology.

[0361] Triptolide is a diterpenoid 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. [ka]

[0362] Triptolide may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0363] In certain embodiments, the compositions comprising triptolide are "ready-to-use" formulations containing triptolide in a dissolved or solubilized form and are intended to be used by itself or with further dilution with an intravenous diluent.

[0364] Thus, in certain embodiments, a pharmaceutical composition comprising triptolide is formulated for parenteral administration, eg, by injection or infusion.

[0365] However, in certain embodiments, a pharmaceutical composition comprising triptolide is formulated for oral administration, such as a tablet or capsule.

[0366] In one embodiment, the use of a combination of panobinostat and triptolide for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and triptolide is administered in the form of an injection or infusion.

[0367] In another embodiment, the use of a combination of panobinostat and triptolide for the treatment of cholangiocarcinoma is one in which both panobinostat and triptolide are administered orally.

[0368] Thus, in some embodiments, panobinostat and triptolide may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and triptolide may be administered in a single dosage form (e.g., tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0369] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and triptolide in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0370] In certain embodiments, the clinical dose of panobinostat in combination with triptolide for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0371] In some embodiments, the clinical dose of triptolide in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as triptolide currently used for other indications. For example, in some embodiments, the dose range of triptolide may be 10 to 200 mg, e.g., 25 to 150 mg daily.

[0372] In certain embodiments, the combination therapy comprises administration of panobinostat and BI2536. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 BI2536 or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0373] BI2536 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.

[0374] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with BI2536, or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0375] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with BI2536 or a pharmaceutically acceptable salt, solvate or hydrate thereof for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0376] In certain embodiments, the panobinostat and BI2536 combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and BI2536 in a single dosage form (eg, a tablet or capsule).

[0377] In certain embodiments, combination therapy of panobinostat and BI2536 may be used to treat extrahepatic CCA.

[0378] In one embodiment, combination therapy of panobinostat and BI2536 is used to treat a subject having a CCA tumor with one or more characteristics specific to the TFK-1 cell line, e.g., one or more genetic markers, growth rate, and / or cell morphology.

[0379] BI2536 is an inhibitor of PLK1 (Polo-like kinase 1) protein, having the structure shown below. The code name "BI2536" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. Pharmaceutically acceptable salts are preferably as defined above. For example, in some embodiments, BI2536 may be in the form of an acid salt, such as a hydrochloride salt. [ka]

[0380] BI2536 may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0381] In certain embodiments, the composition comprising BI2536 is a "ready-to-use" formulation containing BI2536 in a dissolved or solubilized form and is intended to be used by itself or with further dilution with an intravenous diluent.

[0382] Thus, in certain embodiments, a pharmaceutical composition comprising BI2536 is formulated for parenteral administration, eg, by injection or infusion.

[0383] However, in certain embodiments, a pharmaceutical composition comprising BI2536 is formulated for oral administration, eg, as a tablet or capsule.

[0384] In one embodiment, the use of a combination of panobinostat and BI2536 for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and BI2536 is administered in the form of injection or infusion.

[0385] In another embodiment, the use of a combination of panobinostat and BI2536 for the treatment of cholangiocarcinoma is one in which both panobinostat and BI2536 are administered orally.

[0386] Thus, in some embodiments, panobinostat and BI2536 may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and BI2536 may be administered in a single dosage form (e.g., a tablet or capsule) as a combined formulation (i.e., a pharmaceutical composition).

[0387] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and BI2536 in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0388] In certain embodiments, the clinical dose of panobinostat in combination with BI2536 for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0389] In certain embodiments, the clinical dose of BI2536 in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for BI2536 for other indications. For example, in certain embodiments, the dose range of BI2536 can be 1 to 200 mg, e.g., 25 to 150 mg daily.

[0390] In some embodiments, the combination therapy comprises administration of panobinostat and dactolisib. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 dactolisib or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0391] Ductolisib 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.

[0392] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with dactolisib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0393] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with dactolisib or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0394] In certain embodiments, the panobinostat and dactolisib combination product is a fixed-dose product, eg, a pharmaceutical composition, comprising panobinostat and dactolisib in a single dosage form (eg, a tablet or capsule).

[0395] The combination of panobinostat and dactolisib can be used to treat extrahepatic or intrahepatic CCA. In some embodiments, the combination of panobinostat and dactolisib is used to treat extrahepatic CCA.

[0396] In certain embodiments, the combination therapy of panobinostat and dactolisib is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1 cell line, HuCC-T1 cell line, EGI-1 cell line, and / or TFK-1 cell line, preferably the CC-SW-1 cell line and / or the TFK-1 cell line, most preferably the TFK-1 cell line.

[0397] Ductolisib is a phosphoinositide 3-kinase inhibitor (PI3K inhibitor) that also inhibits mTOR. Ductolisib has the structure shown below. The term "ductolisib" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. [ka]

[0398] Ductolisib may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0399] In certain embodiments, the composition containing dacutolisib is a "ready-to-use" formulation containing dacutolisib in a dissolved or solubilized form and is intended for use by itself or upon further dilution with an intravenous diluent.

[0400] Thus, in some embodiments, a pharmaceutical composition comprising dactolisib is formulated for parenteral administration, e.g., by injection or infusion.

[0401] However, in certain embodiments, a pharmaceutical composition comprising dactolisib is formulated for oral administration, e.g., as a tablet or capsule.

[0402] In one embodiment, the use of a combination of panobinostat and dactolisib for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and dactolisib is administered in the form of an injection or infusion.

[0403] In another embodiment, the use of a combination of panobinostat and dactolisib for the treatment of cholangiocarcinoma is one in which both panobinostat and dactolisib are administered orally.

[0404] Thus, in some embodiments, panobinostat and dactolisib may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and dactolisib may be administered in a single dosage form (e.g., a tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0405] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and dactolisib in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0406] In certain embodiments, the clinical dose of panobinostat in combination with dactolisib for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0407] In some embodiments, the clinical dose of dactolisib in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for dactolisib for other indications. For example, in some embodiments, the dose range of dactolisib may be 100 to 1200 mg, e.g., 200 to 800 mg daily.

[0408] In some embodiments, the combination therapy comprises administration of panobinostat and obatoclax. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 obatoclax or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0409] Obatoclax 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.

[0410] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with obatoclax, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in the subject.

[0411] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with obatoclax or a pharmaceutically acceptable salt, solvate or hydrate thereof for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0412] In certain embodiments, the panobinostat and obatoclax combination product is a fixed-dose product, eg, a pharmaceutical composition, that includes panobinostat and obatoclax in a single dosage form (eg, a tablet or capsule).

[0413] In certain embodiments, combination therapy of panobinostat and obatoclax may be used to treat extrahepatic CCA.

[0414] In certain embodiments, the combination therapy of panobinostat and obatoclax is used to treat a subject having a CCA tumor with one or more characteristics specific to the EGI-1 and / or TFK-1 cell lines, e.g., one or more genetic markers, growth rate, and / or cell morphology.

[0415] Obatoclax is an inhibitor of the Bcl-2 family of proteins having the structure shown below. The term "obatoclax" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. The pharmaceutically acceptable salt is preferably obatoclax mesylate. [ka]

[0416] Obatoclax may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention.

[0417] In some embodiments, the composition comprising obatoclax is a "ready-to-use" formulation containing obatoclax in a dissolved or solubilized form and is intended to be used by itself or after further dilution with an intravenous diluent.

[0418] Thus, in some embodiments, a pharmaceutical composition comprising obatoclax is formulated for parenteral administration, eg, by injection or infusion.

[0419] However, in certain embodiments, a pharmaceutical composition comprising obatoclax is formulated for oral administration, eg, as a tablet or capsule.

[0420] In one embodiment, the use of a combination of panobinostat and obatoclax for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and obatoclax is administered in the form of an injection or infusion.

[0421] In another embodiment, the use of a combination of panobinostat and obatoclax for the treatment of cholangiocarcinoma is one in which both panobinostat and obatoclax are administered orally.

[0422] Thus, in some embodiments, panobinostat and obatoclax may be administered in separate dosage forms (e.g., separate tablets or capsules). In some embodiments, panobinostat and obatoclax may be administered in a single dosage form (e.g., tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0423] Thus, formulations (ie pharmaceutical compositions) comprising both panobinostat and obatoclax in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0424] In certain embodiments, the clinical dose of panobinostat in combination with obatoclax for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0425] In certain embodiments, the clinical dose of obatoclax in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for obatoclax for other indications. For example, in certain embodiments, the dose range for obatoclax is 5 to 50 mg / m 2 BSA, e.g., 10–20 mg / m 2 BSA can be administered daily.

[0426] In some embodiments, the combination therapy comprises administration of panobinostat and elesclomol. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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 elesclomol or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0427] Elesclomol 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.

[0428] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, in the manufacture of a combination product with elesclomol, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0429] In another embodiment, the present invention provides panobinostat, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, in the manufacture of a combination product with elesclomol, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0430] In certain embodiments, the panobinostat and elesclomol combination product is a fixed dose, eg, a pharmaceutical composition, that includes panobinostat and elesclomol in a single dosage form (eg, injection or infusion).

[0431] Combination therapy of panobinostat and elesclomol can be used to treat extrahepatic or intrahepatic CCA.

[0432] In certain embodiments, the combination therapy of panobinostat and elesclomol is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate, and / or cell morphology, specific for the CC-SW-1, HuCC-T1, EGI-1, and / or TFK-1 cell lines, preferably the TFK-1 cell line.

[0433] Elesclomol (1-N',3-N'-bis(benzenecarbonothioyl)-1-N',3-N'-dimethylpropanedihydrazide) induces oxidative stress, producing high levels of reactive oxygen species (ROS), such as hydrogen peroxide, in both cancer and normal cells. Elesclomol has the structure shown below. The term "elesclomol" includes pharmaceutically acceptable salts, solvates, and hydrates thereof. In some embodiments, elesclomol is provided in the form of a sodium salt. [ka]

[0434] Elesclomol may be provided as a liquid or solid pharmaceutical composition for use in the methods, compositions and uses of the present invention. Elesclomol is described in WO2013071106, which is incorporated herein by reference.

[0435] In some embodiments, the composition comprising elesclomol is a "ready-to-use" formulation containing elesclomol in dissolved or solubilized form and is intended to be used by itself or after further dilution with an intravenous diluent.

[0436] Thus, in some embodiments, a pharmaceutical composition comprising elesclomol is formulated for parenteral administration, eg, by injection or infusion.

[0437] However, in certain embodiments, a pharmaceutical composition comprising elesclomol is formulated for oral administration, such as a tablet or capsule.

[0438] In one embodiment, the use of a combination of panobinostat and elesclomol for the treatment of cholangiocarcinoma is such that panobinostat is administered orally and elesclomol is administered in the form of an injection or infusion.

[0439] In another embodiment, the use of a combination of panobinostat and elesclomol for the treatment of cholangiocarcinoma is one in which both panobinostat and elesclomol are administered orally.

[0440] Thus, in some embodiments, panobinostat and elesclomol may be administered in separate dosage forms (e.g., tablets or capsules), or in some embodiments, panobinostat and elesclomol may be administered in a single dosage form (e.g., tablet or capsule) as a combined formulation (i.e., pharmaceutical composition).

[0441] Thus, formulations (ie pharmaceutical compositions) containing both panobinostat and elesclomol in the same combination (eg tablet or capsule) for the treatment of cholangiocarcinoma form a further aspect of the present invention.

[0442] In certain embodiments, the clinical dose of panobinostat in combination with elesclomol for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0443] In certain embodiments, the clinical dose of elesclomol in combination with panobinostat for the treatment of cholangiocarcinoma is generally in the same range as that currently used for elesclomol for other indications. For example, in certain embodiments, the dose range of elesclomol is 50 to 300 mg / m 2 BSA, e.g., 100-200 mg / m 2 BSA can be administered daily.

[0444] In some embodiments, the combination therapy comprises administration of panobinostat and docetaxel. Thus, the present invention provides a method of treating cholangiocarcinoma in a subject, 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.

[0445] 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.

[0446] Viewed another way, the present invention provides panobinostat, or a pharmaceutically acceptable salt thereof, as a combination product with docetaxel, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for separate, simultaneous, or sequential use or administration to a subject for use in treating cholangiocarcinoma in a subject.

[0447] In another embodiment, the present invention provides the use of panobinostat or a pharmaceutically acceptable salt thereof in the manufacture of a combination product with docetaxel or a pharmaceutically acceptable salt, solvate or hydrate thereof, for separate, simultaneous or sequential use or administration to a subject for the treatment of cholangiocarcinoma in the subject.

[0448] In certain embodiments, combination therapy of panobinostat and docetaxel is used for the treatment of intrahepatic CCA.

[0449] In certain embodiments, the combination therapy of panobinostat and docetaxel is used for the treatment of a subject having a CCA tumor having one or more characteristics, e.g., one or more genetic markers, growth rate and / or cell morphology, specific for the CC-SW-1 cell line and / or TFK-1 cell line, preferably the CC-SW-1 cell line.

[0450] Docetaxel (N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetylpaclitaxel) is an antimitotic chemotherapy drug that reversibly binds to tubulin with high affinity at a 1:1 stoichiometry. Docetaxel has the structure shown below and is widely available from Actavis, etc. The term "docetaxel" includes its pharmaceutically acceptable solvates and hydrates. In some embodiments, docetaxel is provided as docetaxel trihydrate. [ka]

[0451] 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.

[0452] In certain embodiments, the docetaxel-containing composition is a "ready-to-use" formulation containing docetaxel in a dissolved or solubilized form, and is intended for use by itself or upon further dilution with an intravenous diluent.

[0453] In a preferred embodiment, the pharmaceutical composition comprising docetaxel is formulated for parenteral administration.

[0454] A preferred embodiment of the use of a combination of panobinostat and docetaxel for the treatment of cholangiocarcinoma is to administer panobinostat orally and docetaxel in the form of injection or infusion.

[0455] In certain embodiments, the clinical dose of panobinostat in combination with docetaxel for the treatment of cholangiocarcinoma is generally 5-50 mg, more preferably 10-30 mg, daily or at least twice a week, as defined above.

[0456] In certain embodiments, the clinical dose of docetaxel in combination with panobinostat for the treatment of cholangiocarcinoma will generally be in the same range as that currently used when docetaxel is used for other indications, e.g., 20-200 mg / m 2 Body surface area (BSA), preferably 40-75 mg / m 2 BSA every day.

[0457] The drug substances disclosed herein (i.e., panobinostat and cytotoxic agents) may be in the form of free drugs or their pharmaceutically acceptable salts, solvates, or hydrates according to the present invention. Such salts, solvates, and hydrates are fully described in the literature. Any suitable pharmaceutically acceptable salt, solvate, or hydrate of the drug substances disclosed herein may be used according to the present invention for the treatment of cholangiocarcinoma.

[0458] The preferred form of the drug substance is the form present in the regulatory approved, commercially available drug product.

[0459] The drugs may be administered simultaneously or sequentially. If the drugs are administered sequentially, the interval between drug administrations may vary from minutes to days, depending on the nature of the drug substance and the clinical situation.

[0460] Thus, panobinostat and the additional cytotoxic agent may be used simultaneously, separately, or sequentially. When used simultaneously, they may be administered simultaneously but by a single route or by different routes (e.g., a mixture administered orally or two (or more) formulations administered simultaneously but by different routes, i.e., orally and intravenously). When administered separately, they may be administered simultaneously or sequentially and / or may overlap in timing. In some embodiments, multiple 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 BI2536, panobinostat and combretastatin A4, panobinostat and dactolisib, panobinostat and dapolinad, panobinostat and ispinesib, panobinostat and luminespib, panobinostat and molybrespib, panobinostat and obatoclax, panobinostat and pelitinib, panobinostat and elesclomol, and panobinostat and triptolide.

[0461] In certain embodiments of the invention, panobinostat and / or other cytotoxic agents are administered more than once, for example, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times (e.g., up to 20 times), which may be in one (or each) cycle or multiple cycles combined.

[0462] As used herein, a "cycle" refers to the period during which a particular treatment regimen is applied, generally repeated to provide cyclical treatment. The treatment in each cycle may be the same or different (e.g., different dosages, timing, etc. may be used). The cycle length is 7 to 30 days, e.g., 14- or 21-day cycles. In some embodiments, a cycle may be about 1 to 3 months. Multiple cycles may be used, e.g., at least 2, 3, 4, or 5 cycles, e.g., 6, 7, 8, 9, or 10 cycles (e.g., up to 8, 9, 10, or 20 cycles). Within each cycle, panobinostat and / or other cytotoxic agents may be administered once or more than once, as described hereinabove.

[0463] When a combination drug therapy is administered separately or sequentially, two or more different drugs (e.g., panobinostat and another cytotoxic agent) can be provided as a combination product in which the 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 both formulations and, optionally, instructions for administration.

[0464] When combination drug therapy is administered simultaneously, two or more different drugs (eg, panobinostat and another cytotoxic agent) can be administered together in a single formulation in a so-called combination drug.

[0465] Therefore, another embodiment of the present invention relates to a combination (pharmaceutical composition) comprising panobinostat and one or more cytotoxic agents for the treatment of cholangiocarcinoma. In a preferred embodiment, the one or more cytotoxic agents are selected from dasatinib, topotecan, methotrexate, trametinib, BI2536, combretastatin A4, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, molybresil, obatoclax, pelitinib, triptolide, and combinations thereof. Such combinations can be easily prepared using well-known formulation techniques.

[0466] However, in some embodiments, the different drugs may be administered simultaneously in separate forms, for example, separate tablets.

[0467] Thus, in a further embodiment, the present invention may be considered to provide a kit comprising panobinostat and a cytotoxic agent as defined above, preferably for simultaneous, separate or sequential use for the treatment of cholangiocarcinoma in a patient, wherein preferably said use is as defined herein above.

[0468] In preferred embodiments, the cytotoxic agent is selected from bortezomib, BI2536, carboplatin, cisplatin, combretastatin A4, dactolisib, dapolinad, dasatinib, doxorubicin, docetaxel, elesclomol, gemcitabine, ispinesib, luminespib, methotrexate, molybrespib, obatoclax, pelitinib, SB-743921, topotecan, trametinib, and triptolide, and combinations thereof.

[0469] In further preferred embodiments, the cytotoxic agent is selected from BI2536, carboplatin, cisplatin, combretastatin A4, dactolisib, dapolinad, dasatinib, doxorubicin, docetaxel, elesclomol, ispinesib, luminespib, methotrexate, molybrespib, obatoclax, pelitinib, SB-743921, topotecan, trametinib and triptolide, and combinations thereof.

[0470] In other preferred embodiments, the additional cytotoxic agent is selected from doxorubicin, dactolisib, SB-743921, trametinib, elesclomol, molybreceptor, methotrexate, dapolinad, topotecan, cisplatin, dasatinib, carboplatin, and luminespib.

[0471] In yet another preferred embodiment, the additional cytotoxic agent is selected from carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, molybresib, obatoclax, pelitinib, trametinib and triptolide, preferably carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib.

[0472] In addition to the above-described drug substances and combinations for the treatment of cholangiocarcinoma, the compositions, kits, or therapeutic regimens of the present invention may contain other drugs, which may be other anti-cancer drugs or drugs known to be administered in cancer treatment regimens, such as other cytotoxic agents described herein.

[0473] In some embodiments of the present invention, the subject (patient) may be subjected to other treatments before, simultaneously with, or after the treatment of the present invention.For example, in some embodiments, the subject (patient) may be treated with radiation therapy and / or surgery according to methods known in the art.

[0474] Thus, in certain embodiments, the methods of the present invention may include the further step of treating the subject with radiation therapy and / or surgery. Surgery may include resection of the CCA tumor.

[0475] In certain embodiments, the combination therapy of the present invention can be used as a second-line treatment, i.e., in subjects refractory to gemcitabine-based therapy. Thus, in certain embodiments, the subject to be treated is refractory to gemcitabine-based therapy.

[0476] BSA (body surface area) can be calculated using, for example, the Mosteller method.

number

[0477] Preferred embodiments of the present invention are illustrated in the Examples, where one or more of the parameters or elements used in the Examples can be used as preferred characteristics of the methods described above.

[0478] The invention will now be further described in the following non-limiting examples and with reference to the following figures: [Brief explanation of the drawings]

[0479] [Figure 1] Concentration-response curves for panobinostat in seven cholangiocarcinoma cell lines are shown. A) Intrahepatic cholangiocarcinoma cell line. B) Extrahepatic cholangiocarcinoma cell line and KMCH-1 as a combined cholangiocarcinoma and hepatocellular carcinoma cell line. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0480] [Figure 2] The effects of panobinostat (squares) and bortezomib (triangles) as single agent treatments and the combination of panobinostat with 1.3 nM bortezomib (circles) are shown in the cell line HuCC-T1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0481] [Figure 3]The effects of panobinostat (squares) and carboplatin (triangles) as single agent treatments and in combination with 1000 nM carboplatin (circles) on the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0482] [Figure 4] The effects of panobinostat (squares) and cisplatin (triangles) as single agent treatments and the combination of panobinostat with 100 nM cisplatin (circles) on the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0483] [Figure 5] The effects of panobinostat (squares) and dasatinib (triangles) as single agent treatments and the combination of panobinostat with 5 nM dasatinib (circles) are shown in the cell line TFK-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0484] [Figure 6] The effects of panobinostat (squares) and doxorubicin (triangles) as single agent treatments and in combination with 87 nM or 100 nM doxorubicin (circles) are shown in cell lines (A) HuCCT-1 and (B) TFK-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0485] [Figure 7] The effects of panobinostat (squares) and gemcitabine (triangles) as single agent treatments and in combination with 12 nM or 1000 nM gemcitabine (circles) are shown in cell lines (A) CC-SW-1 and (B) TFK-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0486] [Figure 8] The effects of panobinostat (squares) and methotrexate (triangles) as single agent treatments and in combination with 24 nM methotrexate (circles) on the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0487] [Figure 9] The effects of panobinostat (squares) and trametinib (triangles) as single agent treatments and in combination with 8.8 nM or 1000 nM trametinib (circles) are shown in cell lines (A) HuCCT-1 and (B) TFK-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0488] [Figure 10] The effects of panobinostat (squares) and topotecan (triangles) as single agent treatments and in combination with 24 nM or 120 nM topotecan (circles) are shown in cell lines (A) CC-SW-1 and (C) TFK-1. (B) The effects of topotecan (squares) and panobinostat (triangles) as single agent treatments and in combination with 5.3 nM panobinostat (circles) are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0489] [Figure 11] The effects of panobinostat (squares) and BI2536 (triangles) as single agent treatments and the combination of panobinostat with 490 nM BI2536 (circles) on the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0490] [Figure 12]The effects of panobinostat (squares) and triptolide (triangles) as single agent treatments and the combination of panobinostat with 30 nM triptolide (circles) on the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0491] [Figure 13] The effects of panobinostat (squares) and ductolisib (triangles) as single agent treatments and in combination with 80 or 2 nM ductolisib (circles) are shown in cell lines (A) EGI-1 and (B) TFK-1, respectively. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0492] [Figure 14] (A) The effects of panobinostat (squares) and dapolinade (triangles) as single agent treatments and in combination with 23 nM panobinostat (circles) in the cell line TFK-1; and (B) the effects of dapolinade (squares) and panobinostat (triangles) as single agent treatments and in combination with 5.3 nM panobinostat (circles) in the cell line CC-SW-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0493] [Figure 15] The effects of panobinostat (squares) and obatoclax mesylate (triangles) as single agent treatments and in combination with 16 nM obatoclax mesylate (circles) are shown in the cell line TFK-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0494] [Figure 16]The effects of panobinostat (squares) and SB-743921 (triangles) as single agent treatments and the combination of panobinostat with 6.4 nM SB-743921 (circles) on the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0495] [Figure 17] The effects of panobinostat (squares) and combretastatin A4 (triangles) as single agent treatments and in combination with 1000 nM or 100 nM combretastatin A4 (circles) are shown in cell lines (A) EGI-1 and (B) HuCC-T1, respectively. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0496] [Figure 18] The effects of panobinostat (squares) and ispinesib (triangles) as single agent treatments and the combination of panobinostat with 75 nM ispinesib (circles) in the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0497] [Figure 19] The effects of panobinostat (squares) and molybresib (triangles) as single agent treatments and the combination of panobinostat with 1000 nM molybresib (circles) on the cell line TFK-1 are shown. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0498] [Figure 20] The effects of panobinostat (squares) and luminespib (triangles) as single agent treatments and the combination of panobinostat with 56 nM luminespib (circles) are shown in the cell line TFK-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0499] [Figure 21] The effects of panobinostat (squares) and pelitinib (triangles) as single agent treatments and the combination of panobinostat with 90 nM pelitinib (circles) are shown in the cell line TFK-1. Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines.

[0500] [Figure 22] (A) and (B) show the effects of elesclomol (squares) and panobinostat (triangles) as single-substance treatments and the combination of elesclomol with 5.3 nM or 14 nM panobinostat (circles). (C) and (D) show the effects of elesclomol (squares) and elesclomol (triangles) as single-substance treatments and the combination of panobinostat with 7 nM or 70 nM elesclomol (circles). Cell viability was measured 48 hours after drug addition. IC50 values ​​are highlighted with vertical lines. [Example]

[0501] Experimental method and analytical summary of compounded formulations Various cholangiocarcinoma (CCA) cell lines were used in this drug screening. Table 1 provides details of the culture media and cell numbers used in this experiment. [Table 1]

[0502] After trypsinization and counting, single cells were seeded into Greier 384-well tissue culture-treated polystyrene plates (#781098) in 10 μL of the appropriate medium (see Table 1 for exact cell numbers). Seeded cells were allowed to attach to the plate for 24 hours, after which the appropriate volume of compound was added using an acoustic liquid dispenser (Labcyte Echo 550) to obtain single drug concentrations ranging from 0.1 nM to 1000 nM in a total volume of 25 μL ("dose response"). Wells were filled with 15 μL of the appropriate medium and incubated for 48 hours as described above. After 48 hours of incubation, cells were treated with 25 μL of 0.5x Cell TitreGlo Luminescent Viability Detection Reagent. Cells were incubated in the dark for 10 minutes, and then luminescence was read from the top using autogain on a Synergy Neo2 plate reader.

[0503] Seven cholangiocarcinoma cell lines were used for panobinostat as a single agent treatment. Panobinostat was then tested in combination with 23 other drugs on four cholangiocarcinoma cell lines. Single concentrations of the combination drugs were compared with the IC20 previously determined for the combination drugs tested alone on each of the four cell lines. 20 The cell line TFK-1 was insensitive to some of the tested drugs. IC 20 If values ​​could not be calculated, doses between 100 and 1000 nM were selected for combination studies (applicable to carboplatin, cisplatin, gemcitabine, and trametinib). For HuCCT-1 cells, trametinib was administered at an IC 20 Instead, IC 44 = 8.8 nM.

[0504] Data analysis Cell viability from the Cell TitreGlo assay yielded 576 dose-response curves for each cell line tested (24 monotherapies including DMSO and 552 IC 20 (combination agent in

[0505] Example 1: Effect of panobinostat on seven cholangiocarcinoma cell lines Panobinostat was tested in seven different cholangiocarcinoma cell lines as a single agent treatment compared to DMSO, as detailed in the Methods section above, confirming that all analyzed cell lines showed a response to panobinostat (see Figure 1): five cell lines showed an IC of approximately 100 nM. 50 Two cell lines (CC-SW-1 and EGI-1) showed IC of 12 nM or 36 nM. 50 High sensitivity to panobinostat (IC 50 values ​​in Table 2 and dose-response curves in Figure 1). Although effective as a single agent, panobinostat efficacy is increased in combination with other anti-cancer drugs as described below (Examples 2-22). [Table 2]

[0506] Example 2: Combination of panobinostat and bortezomib in HuCCT-1 cholangiocarcinoma cells The cell line HuCCT-1 was treated with panobinostat and low-dose bortezomib (1.3 nM, IC of single substance curves). 20 and 20 mg / kg of guinea pigs treated with a combination of 20 mg / kg of guinea pigs (20 mg / kg of guinea pigs, ... Indeed, the combination of panobinostat and bortezomib significantly improved the IC of panobinostat as a single-substance treatment. 50 It is more efficient as highlighted by the value of 90 nM, which drops to 51 nM in combination with 1.3 nM bortezomib. It can therefore be concluded that panobinostat shows increased efficacy in combination with bortezomib compared to single agent treatment.

[0507] Example 3: Combination of panobinostat and carboplatin against TFK-1 cholangiocarcinoma cells For experimental details of the combination drug test, see the Methods section above. Interestingly, carboplatin did not affect cell viability of TFK-1 cells as a single agent treatment (Figure 3, triangles). However, when carboplatin was added to panobinostat at a dose of 1000 nM (Figure 3, circles), the IC50 The efficacy of panobinostat increased, as indicated by a decrease in the IL-12 value from 70 nM to 26 nM (Figure 3). Therefore, the combination of panobinostat and carboplatin is expected to show better efficacy than panobinostat alone in treating cholangiocarcinoma.

[0508] Example 4: Combination of panobinostat and cisplatin against TFK-1 cholangiocarcinoma cells Cisplatin had no effect on TFK-1 as a single agent treatment (Figure 4, triangles). However, the addition of 100 nM cisplatin to panobinostat (IC 50 =42 nM; Figure 4, circles) showed higher efficacy than panobinostat alone (IC 50 = 70 nM; squares), see Figure 4. For experimental details of the combination drug test, see the Methods section above. Therefore, the combination of panobinostat and cisplatin is expected to be highly effective in treating cholangiocarcinoma.

[0509] Example 5: Combination of panobinostat and dasatinib against TFK-1 cholangiocarcinoma cell line For experimental details of the combination drug test, see the Methods section above. Dasatinib as a single agent showed only modest efficacy in TFK-1 cells (Figure 5, triangles). Interestingly, low doses of dasatinib (5 nM, IC of the single agent curve) 20 The addition of α-glucan (a dose of α-glucan) enhanced the effect of panobinostat, from an IC50 of 70 nM for panobinostat alone (Figure 5, squares) to 39 nM for the combination. 50 This is explained by the shift in values ​​(Figure 5, circles). Therefore, from these results, it is expected that the combination of panobinostat and dasatinib will have higher efficacy than panobinostat alone for the treatment of cholangiocarcinoma.

[0510] Example 6: Combination of panobinostat and doxorubicin against HuCCT-1 and TFK-1 cholangiocarcinoma cells For experimental details of the combination drug test, see the Methods section above. The cell line HuCCT-1 had IC50 values ​​of 90 nM and 158 nM, respectively. 50The combination of panobinostat and doxorubicin (Figure 6A, circles) demonstrated an IC of 44 nM compared with panobinostat (squares) or doxorubicin (triangles) alone. 50 The values ​​showed a good response, see Figure 6A. The cell line TFK-1 had IC of 70 nM and 123 nM, respectively. 50 The combination of panobinostat and doxorubicin (Figure 6B, circles) demonstrated an IC of 40 nM compared with panobinostat (squares) or doxorubicin (triangles) alone. 50 The values ​​showed a good response, see Figure 6B. Therefore, the combination of panobinostat and doxorubicin is expected to be beneficial in the treatment of cholangiocarcinoma.

[0511] Example 7: Combination of panobinostat and gemcitabine in CC-SW-1 and TFK-1 cholangiocarcinoma cell lines For experimental details of the combination drug test, see the Methods section above. The cell line CC-SW-1 is sensitive to both panobinostat (Figure 7A, squares) and gemcitabine (Figure 7A, triangles), but is particularly sensitive to the combination of panobinostat and 12 nM gemcitabine (Figure 7A, circles). The combination has a lower IC of 3 nM compared to 12 nM for panobinostat alone. 50 The values ​​were shown. Interestingly, the cell line TFK-1 was not sensitive to gemcitabine as a single agent treatment. Nevertheless, the addition of gemcitabine to panobinostat (IC 50 =46 nM) increased efficacy (IC 50 = 70 nM (see Figure 7B). Therefore, the addition of gemcitabine to panobinostat leads to an increased effect on cell viability during cholangiocarcinoma treatment.

[0512] Example 8: Combination of panobinostat and methotrexate in TFK-1 cholangiocarcinoma cells For experimental details of the combination drug test, see the Methods section above. Methotrexate had only a small effect on cell viability in TFK-1 cells (Figure 8, triangles). The addition of 24 nM methotrexate to panobinostat (Figure 8, circles) nevertheless resulted in an IC of 70 nM. 50 an IC of 37 nM compared to panobinostat alone (Figure 8, squares). 50 The results showed increased efficacy in the treatment of cholangiocarcinoma (see Figure 8). Similar efficacy is expected for the treatment of cholangiocarcinoma.

[0513] Example 9: Combination of panobinostat and trametinib in HuCCT-1 and TFK-1 cholangiocarcinoma cell lines For experimental details of the combination drug test, see the Methods section above. Trametinib showed only marginal efficacy in both HuCCT-1 and TFK-1 cells (triangles, Figures 9A and B). 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 due to an IC50 increase from 90 nM for panobinostat alone (Figure 9A, squares) to 42 nM for the combination treatment (circles, Figure 9A). 50 This is indicated by a shift in value. Similarly, combination trametinib and panobinostat treatment reduced IC from 70 nM to 31 nM in TFK-1 cells 50 The reduction in values ​​also indicated a greater effect than panobinostat alone (Figure 9B). Therefore, combined treatment with panobinostat and trametinib is expected to be highly effective in treating cholangiocarcinoma.

[0514] Example 10: Combination of panobinostat and topotecan in CC-SW-1 and TFK-1 cholangiocarcinoma cell lines For experimental details of the combination drug test, see the Methods section above. Cell line CC-SW-1 showed IC50 values ​​of 12 nM and 54 nM, respectively. 50 The combination of panobinostat and fixed-dose topotecan in CC-SW-1 cells (Figure 10A, circles) demonstrated an IC of 5.0 nM compared to panobinostat alone (squares).50 See Figure 10A. Furthermore, the combination of topotecan with a fixed dose of panobinostat in CC-SW-1 cells (Figure 10B, circles) also demonstrated an IC of 37 nM compared to topotecan alone (squares). 50 The values ​​showed a good response, see Figure 10B. The cell line TFK-1 also responded to the combination of panobinostat and topotecan with IC of 70 nM and 373 nM, respectively. 50 The combination of panobinostat and fixed-dose topotecan in TFK-1 cells (Figure 10C, circles) demonstrated an IC of 28 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 10C. Therefore, the combination of panobinostat and topotecan is expected to be beneficial in the treatment of cholangiocarcinoma.

[0515] Example 11: Combination of Panobinostat and BI2536 in TFK-1 cholangiocarcinoma cells For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 had IC50 values ​​of 70 nM and 80 nM, respectively. 50 The combination of panobinostat and BI2536 in TFK-1 cells (Figure 11, circles) demonstrated an IC of 38 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 11. Therefore, the combination of panobinostat and BI2536 is expected to be beneficial in the treatment of cholangiocarcinoma.

[0516] Example 12: Combination of Panobinostat and Triptolide in TFK-1 Cholangiocarcinoma Cells For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 had IC50 values ​​of 70 nM and 15 nM, respectively. 50 The combination of panobinostat and triptolide in TFK-1 cells (Figure 12, circles) showed an IC of 64 nM compared to panobinostat (squares). 50The values ​​showed a good response, see Figure 12. Therefore, the combination of panobinostat and triptolide is expected to be beneficial in the treatment of cholangiocarcinoma.

[0517] Example 13: Combination of panobinostat and ductolisib in EGI-1 and TFK-1 cholangiocarcinoma cell lines For experimental details of the combination drug test, see the Methods section above. The cell line EGI-1 had IC50 values ​​of 99 nM and 99 nM, respectively. 50 The combination of panobinostat and fixed-dose dactolisib in EGI-1 cells (Figure 13A, circles) demonstrated an IC of 34 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 13A. The cell line TFK-1 had IC of 70 nM and 93 nM, respectively. 50 The combination of panobinostat and fixed-dose dactolisib in TFK-1 cells (Figure 13B, circles) resulted in an IC of 30 nM compared to panobinostat alone (squares). 50 The combination of panobinostat and dactolisib showed a favorable response in the cholangiocarcinoma (Fig. 13B). Therefore, the combination of panobinostat and dactolisib is expected to be beneficial in the treatment of cholangiocarcinoma.

[0518] Example 14: Combination of panobinostat and dapolinad in TFK-1 and CC-SW-1 cholangiocarcinoma cell lines For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 had IC50 values ​​of 70 nM and 73 nM, respectively. 50 The combination of panobinostat and fixed-dose dactolisib in TFK-1 cells (Figure 14A, circles) had an IC of 48 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 14A. Furthermore, the cell line CC-SW-1 showed IC values ​​of 12 nM and 8 nM, respectively. 50The combination of dapolinad and fixed-dose panobinostat (Figure 14B, circles) showed a favorable response, with a combined drug sensitivity score (DSS) of 34, compared with a DSS of 17 for dapolinad alone (squares), see Figure 14B. Therefore, the combination of panobinostat and dapolinad is expected to be beneficial for the treatment of cholangiocarcinoma.

[0519] Example 15: Combination of Panobinostat and Obatoclax Mesylate in TFK-1 Cholangiocarcinoma Cells For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 showed IC50 values ​​of 70 nM and 1523 nM, respectively. 50 The combination of panobinostat and fixed-dose obatoclax mesylate in TFK-1 cells (Figure 15, circles) had an IC50 of 52 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 15. Therefore, the combination of panobinostat and obatoclax mesylate is expected to be beneficial in the treatment of cholangiocarcinoma.

[0520] Example 16: Combination of panobinostat and SB-743921 in TFK-1 cholangiocarcinoma cell line For experimental details of the drug combination studies, see the Methods section above. The cell line TFK-1 had IC of 70 nM and 8 nM, respectively. 50 The combination of panobinostat and fixed dose SB-743921 in TFK-1 cells (Figure 16, circles) demonstrated an IC of 37 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 16. Therefore, the combination of panobinostat and SB-743921 is expected to be beneficial in the treatment of cholangiocarcinoma.

[0521] Example 17: Combination of Panobinostat and Combretastatin A4 in EGI-1 and HuCC-T1 Cholangiocarcinoma Cells For experimental details of the combination drug test, see the Methods section above. Cell line EGI-1 had IC50 values ​​of 99 nM and >1000 nM, respectively. 50 The combination of panobinostat and fixed-dose combretastatin A4 in EGI-1 cells (Figure 17A, circles) demonstrated an IC of 52 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 17A. The cell line HuCC-T1 had IC of 90 nM and 11 nM, respectively. 50 The combination of panobinostat and fixed-dose combretastatin A4 in HuCC-T1 cells (Figure 17B, circles) demonstrated an IC of 53 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 17B. Therefore, the combination of panobinostat and combretastatin A4 is expected to be beneficial in the treatment of cholangiocarcinoma.

[0522] Example 18: Combination of Panobinostat and Ispinesib in TFK-1 Cholangiocarcinoma Cells For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 had IC50 values ​​of 70 nM and 11 nM, respectively. 50 The combination of panobinostat and fixed-dose ispinesib in TFK-1 cells (Figure 18, circles) demonstrated an IC of 45 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 18. Therefore, the combination of panobinostat and ispinesib is expected to be beneficial in the treatment of cholangiocarcinoma.

[0523] Example 19: Combination of panobinostat and molybresib in TFK-1 cholangiocarcinoma cells For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 had IC50 values ​​of 70 nM and 181 nM, respectively. 50The combination of panobinostat and fixed dose molybresib in HuCC-T1 cells (Figure 19, circles) demonstrated an IC of 26 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 19. Therefore, the combination of panobinostat and molybresib is expected to be beneficial in the treatment of cholangiocarcinoma.

[0524] Example 20: Combination of panobinostat and luminespib in TFK-1 cholangiocarcinoma cells For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 had IC50 values ​​of 70 nM and 21 nM, respectively. 50 The combination of panobinostat and fixed dose luminespib in TFK-1 cells (Figure 20, circles) had an IC of 11 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 20. Therefore, the combination of panobinostat and luminespib is expected to be beneficial in the treatment of cholangiocarcinoma.

[0525] Example 21: Combination of Panobinostat and Pelitinib in TFK-1 Cholangiocarcinoma Cells For experimental details of the combination drug test, see the Methods section above. The cell line TFK-1 showed IC50 values ​​of 70 nM and 0.08 nM, respectively. 50 The combination of panobinostat and fixed-dose pelitinib in TFK-1 cells (Figure 21, circles) demonstrated an IC of 34 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 21. Therefore, the combination of panobinostat and pelitinib is expected to be beneficial in the treatment of cholangiocarcinoma.

[0526] Example 22: Combination of panobinostat and elesclomol in CC-SW-1, EGI-1, HuCC-T1 and TFK-1 cholangiocarcinoma cell lines For experimental details of the combination drug test, see the Methods section above. Cell line CC-SW-1 had IC50 values ​​of 12 nM and 50 nM, respectively. 50The combination of elesclomol with a fixed dose of panobinostat in CC-SW-1 cells (Figure 22A, circles) resulted in an IC of 19 nM compared to elesclomol alone (squares). 50 The values ​​showed a good response, see Figure 22A. The cell line EGI-1 had IC of 99 nM and 34 nM, respectively. 50 The combination of elesclomol with a fixed dose of panobinostat in EGI-1 cells (FIG. 22B, circles) resulted in an IC of 19 nM compared to elesclomol alone (squares). 50 The values ​​showed a good response, see Figure 22B. The cell line HuCC-T1 had IC of 90 nM and 9 nM, respectively. 50 The combination of panobinostat and fixed-dose elesclomol in HuCC-T1 cells (FIG. 22C, circles) showed an IC of 46 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 22C. The cell line TFK-1 had IC of 70 nM and 35 nM, respectively. 50 The combination of panobinostat and fixed-dose elesclomol in TFK-1 cells (Figure 22D, circles) demonstrated an IC of 24 nM compared to panobinostat alone (squares). 50 The values ​​showed a good response, see Figure 22D. Therefore, the combination of panobinostat and elesclomol is expected to be beneficial in the treatment of cholangiocarcinoma.

[0527] Example 23: Capsules containing panobinostat and dasatinib Panobinostat (99% purity) can be purchased from Shandong Sunrise 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.

[0528] Capsules containing panobinostat and dasatinib were prepared as follows. component Panobinostat lactate (equivalent to 15g panobinostat) Dasatinib monohydrate (equivalent to 50g dasatinib) Magnesium stearate 1g 50g mannitol Microcrystalline cellulose (up to 500g) The above ingredients were mixed volumetrically in a mixer and filled into 1000 hard gelatin capsules size 0. Each capsule contains 15 mg panobinostat and 50 mg dasatinib.

[0529] Example 24: Pharmaceutical product containing two different formulations Panobinostat (99% purity) can be purchased from China's Shandong Sunrise Technology Co., Ltd. Alternatively, panobinostat lactate can be produced from panobinostat and lactic acid according to WO2007146716, which is incorporated herein by reference. Capsules similar to Farydak 15 mg (Novartis) are prepared. The capsules are packaged in blisters (6 capsules / blister). Dasatinib monohydrate (≧99.0% purity) can be purchased from Beijing Yibai Biotechnology Co., Ltd., China. Tablets similar to Sprycel 50 mg (Bristol-Myers Squibb) are prepared. Tablets are packaged in blisters (6 tablets / blister) The blisters (5 tablet blisters and 5 capsule blisters) are packed in a pharmaceutical package along with a package insert.

[0530] Example 25: Reduced toxicity of panobinostat in normal bile duct cells in combination with cytotoxic agents compared to panobinostat monotherapy The effects of various cytotoxic agents on the toxicity of panobinostat in normal bile duct cells were examined. The cell line H69 (CVCL_8121) was used in the above experimental method, and the cells were treated with IC 20 The IC of panobinostat (first-line drug) on ​​these cells when used in combination with a cytotoxic agent (second-line drug) when added at concentrations 50 value was determined. The effect of secondary drugs was evaluated by the IC of panobinostat alone. 50 -IC in combination with panobinostat 50 Delta IC, calculated as 50 Measurements were used to quantify the effect of panobinostat on the cytotoxicity of panobinostat. Positive values ​​indicate that the combination is more toxic than panobinostat alone. The greater the difference, the more toxic the combination. Negative delta IC 50 indicates that the combination is less toxic than the monotherapies. The results are shown below. [Table 3] The results show that the test secondary drugs reduce the toxicity of panobinostat in normal cholangiocytes.

[0531] Example 26: Therapeutic index of panobinostat in combination with other cytotoxic agents compared to panobinostat monotherapy The therapeutic index of various panobinostat combination therapies was determined by comparing the effects of combinations and monotherapy (i.e., panobinostat alone) in normal cholangiocytes (cell line H-69) and various CCA cell lines, as described above. Using the experimental method described above, IC 20 The IC of panobinostat (first-line drug) on ​​these cells when used in combination with a cytotoxic agent (second-line drug) when added at concentrations 50 value was determined. Therapeutic index (TI) is the IC 50 IC in CCA cell lines 50 A TI greater than 1 indicates that the treatment is effective in reducing the viability of CCA cells relative to normal cells. A high TI, e.g., 1.5 or greater, indicates that there is a large difference in efficacy between normal and cancer cells, i.e., the treatment exhibits 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 (IC 50 Values ​​in nM).

[0532] [Table 4] [Table 5] [Table 6] [Table 7]

[0533] These results indicate that the tested panobinostat combination therapy is particularly effective against CCA tumors that share characteristics with the CC-SW-1 cell line. However, the data identify combinations that are effective for other cell lines. For example, the combination of trametinib and doxorubicin is particularly effective with trametinib and doxorubicin. The combination of trametinib and topotecan is particularly effective with trametinib and topotecan. The combination of trametinib and carboplatin is particularly effective with trametinib and carboplatin. The combination of dasatinib, methotrexate, dactolisib, topotecan, and trametinib is particularly effective with dasatinib, methotrexate, dactolisib, topotecan, and trametinib.

[0534] Example 27: IC of panobinostat in CCA cell lines 50 Panobinostat combination therapy to regulate Using the experimental data described here, we identified cytotoxic agents that were particularly effective in enhancing the effects of panobinostat in CCA cells. 50 Determined by determining where the positive delta IC 50 The higher the value, the more effective the combination. The table below shows the absolute delta IC 50 Shown are (nM) and relative changes as percentages.

[0535] [Table 8]

[0536] The table below shows that not all combinations are effective in all cell lines, i.e., some combinations have negative delta IC 50 As indicated by the values, it negatively impacts the toxicity of panobinostat in certain cell lines. [Table 9]

[0537] Example 28: Determination of the combination index of panobinostat and elesclomol Data normalization and curve fitting Cell viability from the CTG assay was measured by plotting X dose-response curves (Y monotherapy with DMSO and IC) for each cell line tested. 20 The results were Z combinations of drugs at 100 μM. Viability data for each plate were normalized to the average of eight replicates of 0.1% DMSO and seven replicates of 100 μM benzethonium chloride (BzCl). BzCl serves as a cell killing control, accounting for the background signal from dead cells in the CTG luminescence assay. Raw luminescence data were normalized according to the following formula:

number

number

[0538] Synergy Score Calculation Synergy scores were calculated for each combination drug, based on the monotherapy dose response versus the combination dose response, according to the Loewe additivity, Bliss independence, and zero interaction potential (ZIP) method. These methods calculate predicted responses based on the monotherapy responses of the drugs used in the combination. The measured responses of the combination were then subtracted from these predicted responses to derive a synergy score for each test concentration; a positive score indicates synergy, while a negative score indicates antagonism.

[0539] Loewe additivity The Loewe additivity value was calculated using the specified method. For each drug in the combination, the predicted response was calculated from the response of the 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 values ​​at the measured concentrations were subtracted from this average to derive the synergy score.

number

number

[0540] Bliss Synergy Predicted responses derived using the Bliss model were calculated by multiplying the monotherapy responses of each drug at each concentration tested in combination, and the responses measured at these concentrations were then subtracted from these predicted values ​​to derive a synergy score.

number

number

[0541] ZIP Synergy Predicted responses derived using the ZIP model were calculated according to the Bliss method described above. The observed responses for the combinations were fitted to a log-logistic function, and the curve maxima were determined to be the IC at the relevant dose. 20 The corresponding response of the drug monotherapy was set. These fitted combination values ​​were then subtracted from the predicted response to derive a synergy score.

number

number

[0542] The combination index for panobinostat and elesclomol was determined as described herein. The table below shows that this combination is synergistic in three cell lines, i.e., exhibits a combination index of less than 1. [Table 10]

[0543] Example 29: Genome sequencing of cell lines Whole genome sequencing was performed on the cell lines used here to identify genetic markers (mutations) that are cell line specific and can be predicted to occur in CCA tumors. The table below shows the markers in the cell lines linked to predictive, prognostic, diagnostic, and predisposing biomarkers in the CIViC database and the Cancer Biomarkers Database, coding variants found in known cancer mutation hotspots predicted as cancer driver mutations or curated as etiological, and coding variants found in oncogenes or tumor suppressor genes.

[0544] [Table 11] [Table 12]

[0545] Example 30: Xenograft study in mice cell culture Normal human bile duct cell line (H69) and various human cholangiocarcinoma cell lines (HuCCT, CC-SW1, EGI-1 and TFK-1) are cultured according to standard conditions.

[0546] Mouse experiments All mouse experiments are performed according to protocols approved by the Ethics Committee for the Use of Animals in Research (Norway). Animals are kept in cages in a temperature-controlled environment. Animals have free access to standard chow and water. The light / dark cycle is 12 h / 12 ​​h. Cell suspensions are injected subcutaneously into nude mice. Tumor growth was confirmed 10 days after administration of the cell suspension. Mice were divided into five groups (10 animals per group): Group 1 received no active treatment, Group 2 received Drug A, Group 3 received Drug B, Group 4 received the combination of Drug A and Drug B, and Group 5 received gemcitabine-based combination therapy. All animals had free access to food and water. If the drug is an approved drug by authorities, the drug will be administered at the same dose (per kg) and frequency of administration, and in the same manner, as used in hospitals for the treatment of other cancer diseases. The highest dose and most frequent administration will be used. If the drug is an experimental drug (i.e., not currently approved), the drug will be administered at the same dose (per kg) and frequency of administration, and in the same manner, as used in prior art documents for cancer treatment. Tumor volume is measured weekly throughout the treatment period. Some mice undergo ultrasound and / or MRI examinations to track tumor growth during the treatment period. After 50 days, mice are anesthetized and sacrificed according to standard procedures. Tumors are excised, weighed, and stored in a freezer for further analysis. Results are expected to show that some combinations are highly effective in treating human cholangiocarcinoma in xenograft nude mouse models. In vivo efficacy is expected to correlate well with in vitro cell line efficacy.

[0547] Example 31: Clinical protocol of combination therapy using drug A and drug B as second-line treatment in patients with cholangiocarcinoma A single-arm, open-label, non-randomized, exploratory, multicenter pilot study. Drug A and Drug B are drugs approved by regulatory authorities for other cancer indications.

[0548] 30 participants Admission Criteria: Patients who have been definitively diagnosed with cholangiocarcinoma by histology or cytology Radiographically measurable disease (per RECIST v1.1) Patients who have been previously treated with gemcitabine-based first-line therapy ●Age: 18-80 years old, male or female Women taking contraceptives, if appropriate

[0549] Exclusion criteria: Breastfeeding or pregnant women Severe cardiac dysfunction High blood pressure (systolic ≥ 150mmHg or diastolic ≥ 100mmHg) Hepatitis C and / or human immunodeficiency virus (HIV) and / or Covid-19 positive Primary sclerosing cholangitis and / or inflammatory bowel disease and / or autoimmune disease Aggressive drug treatment for systemic infection - History of allergy or severe adverse events to drugs with the same mechanism of action as the drug or compounded drug in the combination History of substance abuse, including alcohol and / or drug abuse Organ dysfunction Absolute neutrophil count (ANC) < 1,000 / mm 3 [1.0×109 / L] 〇Platelet <75,000 / mm 3 [75×109 / L] Hemoglobin < 109.0 g / dL Total bilirubin > 1.5 x ULN Aspartate aminotransferase / glutamic oxaloacetic transaminase / GOT (AST / SGOT) and alanine aminotransferase / glutamine pyruvate transaminase / GPT (ALT / SGPT) > 2.5 × ULN (AST and ALT > 5 × upper limit of normal (ULN) in the presence of liver metastases) Serum creatinine > 1.5 x ULN and calculated or measured creatinine clearance < 45 mL / min Inorganic phosphorus outside the normal limit Total and ionized serum calcium outside normal limits Other protocol-defined inclusion / exclusion criteria may apply

[0550] dosage The drugs are administered individually at 50% of the approved maximum tolerated dose used in the treatment of other forms of cancer. The individual drugs are administered in the same manner and with the same frequency as the drugs used in other indications. Preferably, the two drugs are administered together.

[0551] period Up to 24 months for each patient

[0552] Evaluation items Primary: Objective response rate (ORR) [time frame: up to 24 months] Defined as the proportion of participants in each cohort who achieved a complete response (CR) or partial response (PR) based on Response Evaluation Criteria in Solid Tumors Version 1.1 (RECIST v1.1)

[0553] Secondary: Progression-free survival (PFS) [time frame: up to 24 months] Median progression-free survival (MPFS) was defined as the time from first dose to disease progression (per RECIST v1.1) or death (whichever occurred first) in each cohort. ●DOR (Duration of Response) [Timeframe: up to 24 months] Defined as the time from the date of first assessment of CR or PR to first disease progression (per RECIST v1.1) or death (whichever occurs first) in each cohort Best overall response [time frame: up to 24 months] Best overall response will be summarized by the proportion of patients with a best overall response of PR, CR, stable disease (SD), or PD. Disease control rate (DCR) [time frame: up to 24 months] Defined as the proportion of participants achieving a best overall response of CR, PR, or stable disease per RECIST v1.1 Overall survival (OS) [time frame: up to 24 months] Defined as the time from first study drug administration to death from any cause in each cohort median overall survival Number of treatment-related adverse events [time frame: up to 24 months] Adverse events and severity, type and frequency reported for the first time after the first dose of study drug / treatment or related to a worsening of pre-existing conditions ●Quality of Life - Quality of Life Analysis Format: https: / / www.eortc.org / app / uploads / sites / 2 / 2018 / 08 / Specimen-QLQ-C30-English.pdf

[0554] Example 32: Clinical protocol of combination therapy using drug A and drug B versus gemcitabine and cisplatin treatment in patients with cholangiocarcinoma Two-arm, double-blind, randomized, clinical Phase III study, multicenter Drugs X and Y are approved by regulatory authorities for other cancer indications 80 participants Arm A: Combination treatment using Drug A and Drug B (40 participants) Arm B: Gemcitabine (1000 mg / m) administered according to regulatory approved dosing 2 ) and cisplatin at a regulatory approved dose (25 mg / m 2 ) combination

[0555] Admission Criteria: Patients who have been definitively diagnosed with cholangiocarcinoma by histology or cytology Radiographically measurable disease (per RECIST v1.1) ●Age: 18-80 years old, male or female Women taking contraceptives, if appropriate

[0556] Exclusion criteria Breastfeeding or pregnant women Severe cardiac dysfunction High blood pressure (systolic ≥ 150mmHg or diastolic ≥ 100mmHg) Hepatitis C and / or human immunodeficiency virus (HIV) and / or Covid-19 positive Primary sclerosing cholangitis and / or inflammatory bowel disease and / or autoimmune disease Aggressive drug treatment for systemic infection - History of allergy or severe adverse events to drugs with the same mechanism of action as the drug or compounded drug in the combination A history of substance abuse, including alcohol and drug abuse Organ dysfunction Absolute neutrophil count (ANC) < 1,000 / mm 3 [1.0×109 / L] 〇Platelet <75,000 / mm 3 [75×109 / L] Hemoglobin < 109.0 g / dL Total bilirubin > 1.5 x ULN Aspartate aminotransferase / glutamic oxaloacetic transaminase / GOT (AST / SGOT) and alanine aminotransferase / glutamine pyruvate transaminase / GPT (ALT / SGPT) > 2.5 × ULN (AST and ALT > 5 × upper limit of normal (ULN) in the presence of liver metastases) Serum creatinine > 1.5 x ULN and calculated or measured creatinine clearance < 45 mL / min Inorganic phosphorus outside the normal limit Total and ionized serum calcium outside normal limits Other protocol-defined inclusion / exclusion criteria may apply

[0557] dosage The results of exploratory studies form the basis for dosing the drugs in the combination. Without appropriate guidance. The drugs in the combination are individually administered at 50% of the approved maximum tolerated dose used in the treatment of other forms of cancer. The individual drugs are administered in the same manner and with the same frequency as the drugs used in other indications. Preferably, the two drugs are administered together.

[0558] period 24 months for each patient

[0559] Evaluation items Primary: Objective response rate (ORR) [time frame: up to 24 months] Defined as the proportion of participants in each cohort who achieved a complete response (CR) or partial response (PR) based on Response Evaluation Criteria in Solid Tumors Version 1.1 (RECIST v1.1)

[0560] Secondary: Progression-free survival (PFS) [time frame: up to 24 months] Median progression-free survival (MPFS) was defined as the time from first dose to disease progression (per RECIST v1.1) or death (whichever occurred first) in each cohort. ●DOR (Duration of Response) [Timeframe: up to 24 months] Defined as the time from the date of first assessment of CR or PR to first disease progression (per RECIST v1.1) or death (whichever occurs first) in each cohort Best overall response [time frame: up to 24 months] Best overall response will be summarized by the proportion of patients with a best overall response of PR, CR, stable disease (SD), or PD. Disease control rate (DCR) [time frame: up to 24 months] Defined as the proportion of participants achieving a best overall response of CR, PR, or stable disease per RECIST v1.1 Overall survival (OS) [time frame: up to 24 months] Defined as the time from first study drug administration to death from any cause in each cohort Median overall survival number. Number of treatment-related adverse events [time frame: up to 24 months] Adverse events and severity, type and frequency reported for the first time after the first dose of study drug / treatment or related to a worsening of pre-existing conditions ●Quality of Life - Quality of Life Analysis Format: https: / / www.eortc.org / app / uploads / sites / 2 / 2018 / 08 / Specimen-QLQ-C30-English.pdf

[0561] Example 33: Reference Example - Analysis of the Effect of Combination Therapy Containing Gemcitabine and Cisplatin The combination of gemcitabine and cisplatin is currently a common treatment for cholangiocarcinoma (see Legemiddelhandboka, https: / / www.legemiddelhandboka.no / T2.2.1.4 / Galleveiscancer and Juan Valle et al, Annals of Oncology 25: 391-398, 2014). This 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. [Table 13] Gemcitabine monotherapy is one of the preferred treatments for cholangiocarcinoma. Gemcitabine monotherapy data in CC-SW-1, EGI-1, HuCCT, and TFK-1 showed IC values ​​of 1.0 vs. ... 50 The values ​​are much lower than those of all cholangiocarcinoma cancer cell lines. The therapeutic indexes are 0.4, 0.4, 0.4, and 0.2, respectively. This indicates that gemcitabine is not a good treatment for cholangiocarcinoma. The combination of gemcitabine and cisplatin is also a preferred clinical treatment for cholangiocarcinoma. The combination index data for the cell lines CC-SW-1 and TFK-1 show some improvement in the therapeutic index. However, the addition of cisplatin to gemcitabine in the cell line EGI-1 negates the effect of gemcitabine. The combination does not affect the therapeutic effect in the cell line HuCCT.

[0562] Example 34: Panobinostat Combination Therapy Shows Synergy in at Least One CCA Cell Line The table below shows whether the panobinostat combination therapy exhibits synergy in at least one CCA cell line. [Table 14]

[0563] In summary, the inventors have conducted extensive testing of anti-cancer drugs and their combinations. In this regard, thousands of known compounds have been reported to have some activity against one or more forms of cancer. To arrive at the results presented here, the inventors first selected suitable compounds, including 384 compounds. Such a compound library yields over 120,000 different combinations containing two substances. After extensive testing of single compounds and combinations of compounds, the inventors identified 20 combinations that showed good efficacy against at least one of the cell lines tested here. This represents approximately 0.02% of the theoretical number of combinations based on the initial selection of 384 different compounds. The present invention further includes the following aspects. 1. A product comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent for use in treating cholangiocarcinoma in a subject, wherein the cytotoxic agent is selected from carboplatin, BI2536, cisplatin, combretastatin A4, dactolisib, dapolinad, dasatinib, doxorubicin, docetaxel, elesclomol, ispinesib, luminespib, methotrexate, molybrespib, obatoclax, pelitinib, SB-743921, topotecan, trametinib, triptolide, or any one or more of their pharmaceutically acceptable salts, solvates, or hydrates. 2. A product for use according to item 1, wherein the panobinostat or a pharmaceutically acceptable salt thereof and the cytotoxic agent are for separate, simultaneous or sequential use or administration. 3. A product for use according to paragraph 1 or 2, wherein the cytotoxic agent is selected from any one or more of carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. 4. A product for use according to any one of items 1 to 3, wherein the bile duct cancer is intrahepatic bile duct cancer. 5. A product for use according to any one of items 1 to 3, wherein the bile duct cancer is extrahepatic bile duct cancer. 6. A product for use according to any one of items 1 to 5, wherein the panobinostat or a pharmaceutically acceptable salt thereof is provided in a pharmaceutical composition together with a pharmacologically acceptable excipient. 7. A product for use according to paragraph 6, wherein the pharmaceutical composition is formulated for oral administration. 8. A product for use according to paragraph 6 or 7, wherein the pharmaceutical composition is in the form of a tablet or capsule. 9. The product for use according to any one of paragraphs 6 to 8, wherein the pharmaceutical composition comprises an additional cytotoxic agent selected from any one or more of dasatinib, methotrexate, topotecan, trametinib, BI2536, combretastatin A4, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, molybresib, obatoclax, pelitinib, triptolide, or a pharmaceutically acceptable salt thereof. 10. The product for use according to any one of paragraphs 6 to 9, wherein the additional cytotoxic agent is selected from any one or more of dasatinib, methotrexate, topotecan, trametinib, BI2536, combretastatin A4, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, molybresib, obatoclax, pelitinib, triptolide, or a pharmaceutically acceptable salt thereof. 11. A product for use according to any of paragraphs 1 to 7, wherein the cytotoxic agent is formulated for parenteral administration. 12. A product for use according to paragraph 11, wherein the cytotoxic agent is formulated for administration by injection or infusion, preferably intravenous injection or infusion. 13. A product for use according to paragraph 11 or 12, wherein the cytotoxic agent for parenteral administration is selected from any one or more of carboplatin, BI2536, cisplatin, combretastatin A4, dactolisib, dapolinad, doxorubicin, docetaxel, elesclomol, ispinesib, luminespib, methotrexate, molybdesib, obatoclax, pelitinib, SB-743921, topotecan, triptolide, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. 14. A kit comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent selected from carboplatin, BI2536, cisplatin, combretastatin A4, dactolisib, dapolinad, dasatinib, doxorubicin, docetaxel, elesclomol, ispinesib, luminespib, methotrexate, molybreceptor, obatoclax, pelitinib, SB-743921, topotecan, triptolide, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. 15. The kit of paragraph 14, wherein the panobinostat or a pharmaceutically acceptable salt thereof and the cytotoxic agent are for simultaneous, separate or sequential use for treating cholangiocarcinoma in a subject. 16. The kit according to item 14 or 15, wherein the panobinostat or a pharmaceutically acceptable salt thereof, the cytotoxic agent, or the cholangiocarcinoma is defined in any one of items 4 to 13. 17. A pharmaceutical composition comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent selected from dasatinib or a pharmaceutically acceptable salt thereof, methotrexate or a pharmaceutically acceptable salt thereof, topotecan or a pharmaceutically acceptable salt thereof, BI2536 or a pharmaceutically acceptable salt thereof, combretastatin A4 or a pharmaceutically acceptable salt thereof, dactolisib or a pharmaceutically acceptable salt thereof, dapolinad or a pharmaceutically acceptable salt thereof, elesclomol or a pharmaceutically acceptable salt thereof, ispinesib or a pharmaceutically acceptable salt thereof, luminespib or a pharmaceutically acceptable salt thereof, molybresilin or a pharmaceutically acceptable salt thereof, obatoclax or a pharmaceutically acceptable salt thereof, pelitinib or a pharmaceutically acceptable salt thereof, triptolide or a pharmaceutically acceptable salt thereof, and combinations thereof. 18. The pharmaceutical composition of paragraph 17, wherein the composition is formulated for oral administration. 19. The pharmaceutical composition of paragraph 17 or 18, wherein the composition is in the form of a tablet or capsule. 20. The pharmaceutical composition of any one of items 17 to 19, for use in treating bile duct cancer in a subject. 21. Panobinostat or a pharmaceutically acceptable salt thereof for use in combination with a cytotoxic agent selected from any one or more of carboplatin, BI2536, cisplatin, combretastatin A4, dactolisib, dapolinad, dasatinib, doxorubicin, docetaxel, elesclomol, ispinesib, luminespib, methotrexate, molybresilient, obatoclax, pelitinib, SB-743921, topotecan, trametinib, triptolide, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, for use in treating cholangiocarcinoma in a subject. 22. Panobinostat or a pharmaceutically acceptable salt thereof for use according to paragraph 21, wherein the panobinostat or a pharmaceutically acceptable salt thereof and the cytotoxic agent are for separate, simultaneous or sequential use or administration. 23. Panobinostat or a pharmaceutically acceptable salt thereof for use in item 21 or 22, wherein the cytotoxic agent is selected from any one or more of carboplatin, cisplatin, dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

Claims

1. 1. A product comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent for use in treating cholangiocarcinoma in a subject, wherein the cytotoxic agent is selected from carboplatin, BI2536, combretastatin A4, dactolisib, dapolinad, dasatinib, doxorubicin, docetaxel, elesclomol, ispinesib, luminespib, methotrexate, molybresil, obatoclax, pelitinib, SB-743921, topotecan, trametinib, triptolide, or any one or more of their pharmaceutically acceptable salts, solvates, or hydrates.

2. 10. The product for use of claim 1, wherein the panobinostat or a pharmaceutically acceptable salt thereof and the cytotoxic agent are for separate, simultaneous or sequential use or administration.

3. 3. A product for use according to claim 1 or 2, wherein the cytotoxic agent is selected from any one or more of BI2536, combretastatin A4, dactolisib, dapolinad, dasatinib, elesclomol, ispinesib, luminespib, methotrexate, molybreceptor, obatoclax, pelitinib, SB-743921, topotecan, trametinib, triptolide, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

4. 3. The product for use of claim 1 or 2, wherein the cytotoxic agent is selected from any one or more of dasatinib, doxorubicin, docetaxel, methotrexate, topotecan, trametinib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

5. 3. The product for use of claim 1 or 2, wherein the cytotoxic agent is selected from any one or more of dasatinib, methotrexate, topotecan, trametinib, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

6. The product for use according to any one of claims 1 to 5, wherein the cholangiocarcinoma is intrahepatic cholangiocarcinoma.

7. The product for use according to any one of claims 1 to 5, wherein the cholangiocarcinoma is extrahepatic cholangiocarcinoma.

8. A product for use according to claims 1 to 7, wherein the product is formulated for oral administration.

9. A product for use according to claim 8, wherein the product is in the form of a tablet or capsule.

10. A product for use according to claim 8 or 9, comprising a cytotoxic agent selected from any one or more of dasatinib, methotrexate, topotecan, trametinib, BI2536, combretastatin A4, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, molybresib, obatoclax, pelitinib, triptolide or a pharmaceutically acceptable salt thereof.

11. 8. The product for use of any of claims 1 to 7, wherein the cytotoxic agent is formulated for parenteral administration.

12. 12. The product for use of claim 11, wherein the cytotoxic agent is formulated for administration by injection or infusion.

13. A product for use according to claim 11 or 12, wherein the cytotoxic agent is formulated for administration by intravenous injection or infusion.

14. 14. The product for use of any of claims 1 to 13, wherein the cytotoxic agent for parenteral administration is selected from any one or more of carboplatin, BI2536, combretastatin A4, dactolisib, dapolinad, doxorubicin, docetaxel, elesclomol, ispinesib, luminespib, methotrexate, molybreceptor, obatoclax, pelitinib, SB-743921, topotecan, triptolide, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

15. 14. The product for use of any of claims 1 to 13, wherein the cytotoxic agent for parenteral administration is selected from any one or more of BI2536, combretastatin A4, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, methotrexate, molybreceptor, obatoclax, pelitinib, SB-743921, topotecan, triptolide, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

16. A kit for use in treating cholangiocarcinoma in a subject, comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent selected from any one or more of carboplatin, BI2536, combretastatin A4, dactolisib, dapolinad, dasatinib, elesclomol, ispinesib, luminespib, methotrexate, molybreceptor, obatoclax, pelitinib, SB-743921, topotecan, triptolide or a pharmaceutically acceptable salt, solvate or hydrate thereof.

17. 17. The kit for use of claim 16, wherein the cytotoxic agent is selected from any one or more of BI2536, combretastatin A4, dactolisib, dapolinad, dasatinib, elesclomol, ispinesib, luminespib, methotrexate, molybreceptor, obatoclax, pelitinib, SB-743921, topotecan, triptolide, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

18. 18. The kit for use of claim 16 or 17, wherein the cytotoxic agent is selected from any one or more of dasatinib, methotrexate, topotecan, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

19. A pharmaceutical composition for use in treating cholangiocarcinoma in a subject, comprising panobinostat or a pharmaceutically acceptable salt thereof and a cytotoxic agent selected from dasatinib, methotrexate, topotecan, BI2536, combretastatin A4, dactolisib, dapolinad, elesclomol, ispinesib, luminespib, molybresib, obatoclax, pelitinib, triptolide, or a pharmaceutically acceptable salt of any of the foregoing.

20. 20. The pharmaceutical composition for use of claim 19, wherein the cytotoxic agent is selected from dasatinib or a pharmaceutically acceptable salt thereof, methotrexate or a pharmaceutically acceptable salt thereof, and topotecan or a pharmaceutically acceptable salt thereof.

21. 21. The pharmaceutical composition for use of claim 19 or 20, wherein the composition is formulated for oral administration.

22. 22. The pharmaceutical composition for use according to any of claims 19 to 21, wherein the composition is in the form of a tablet or capsule.

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