KR20190122251A - How to treat cancer - Google Patents

Abstract

The present invention relates to a method of treating or preventing cancer in a subject in need thereof comprising administering a therapeutically effective amount of a compound of the invention.

Description

How to treat cancer

Cancer is primarily characterized by an increase in the number of abnormal cells derived from a given normal tissue, invasion of adjacent tissues by abnormal cells, or lymphatic system or blood-mediated spread (metastasis) to local lymph nodes and distant sites of malignant cells. Clinical data and molecular biology studies suggest that cancer begins with minor pre-tumoral changes and is a multi-step process that can progress to tumors under certain conditions.

Primary liver cancer is one of the most common forms of cancer in the world. There are two main types of liver cancers: hepatocellular carcinoma (HCC), also known as malignant liver cancer, and gallbladder cell carcinoma, also known as cholangiocarcinoma (CCA). HCC is the most common form of primary liver cancer, which occurs in liver cells. HCC occurs mainly in males and patients with sclerosis. HCC is one of the most common cancers in the world and the third most common cause of cancer-related deaths. This disease is often diagnosed late in the course of clinical manifestations. As a result, only 10% to 15% of patients are candidates for radical surgery. For most HCC patients, systemic chemotherapy or supportive therapy is the central treatment option.

In contrast, gallbladder cell carcinoma or cholangiocarcinoma occurs in small bile duct epithelial cells (ie, bile duct cells) in the liver. This type of cancer is more common in women. The incidence of cholangiocarcinoma (CCA) is increasing worldwide and is already the second most common primary liver cancer. CCA patients show poor results due to late diagnosis and the refractory nature of these tumors.

To date, the number of drugs that can effectively treat cancers such as HCC and / or CCA is limited. For example, patients with metastatic hepatocellular carcinoma or hepatocellular carcinoma that have failed localized treatment usually survive only 3 to 4 months. Metastatic hepatocellular carcinoma or hepatocellular carcinoma in which local treatment has failed is mainly subject to systemic therapy. The use of high dose tamoxifen or EA-PFL (etoposide, adriamycin, cisplatin, fluorouracil and leucovorin) in combination with doxorubicin, doxorubicin is an effective example. Relief rates of these drugs can achieve levels of 15% to 30%. However, patients with hepatocellular carcinoma usually develop complications of liver cirrhosis and other complications (such as leukopenia, thrombocytopenia or liver dysfunction), so they cannot receive systemic chemotherapy. In addition, most chemotherapeutic agents have limited effects and could not significantly improve patient survival. Despite ongoing efforts, the detrimental clinical course of most cancer patients highlights the need for more effective chemotherapy.

Cholangiocarcinoma is a relatively rare tumor classified as adenocarcinoma (cancer that forms a gland or secretes significant amounts of mucin). The West has an annual incidence of 1-2 cases per 100,000 cases, but bile duct cancer has increased worldwide over the past decades. Cholangiocarcinoma is considered an incurable and rapidly fatal cancer if both the primary tumor and any metastasis cannot be completely removed surgically. There is potentially no cure except for surgery, but most people have advanced disease at the time of onset, and surgery is impossible at diagnosis. People with cholangiocarcinoma are generally not treated but managed with chemotherapy, radiation therapy and other palliative care measures.

The present invention addresses this need. It is therefore an object of the present invention to provide an improved method of treatment or prevention of cancers such as hepatocellular carcinoma and cholangiocarcinoma.

The present application is directed to a method of treating or preventing cancer in a subject in need thereof comprising administering a therapeutically effective amount of a Panesoid X receptor (FXR) agonist. In one embodiment, the FXR agonist is Compound (1) or Compound (2):

,

,

Or a pharmaceutically acceptable salt or amino acid conjugate thereof.

In one embodiment, the cancer is selected from the group consisting of hepatocellular carcinoma, pancreatic cancer, kidney cancer, prostate cancer, esophageal cancer, breast cancer, stomach cancer, kidney cancer, salivary gland cancer, ovarian cancer, uterine cancer, bladder cancer, and lung cancer. In one embodiment, the cancer is hepatocellular carcinoma. In one embodiment, the cancer is pancreatic cancer. In one embodiment, the cancer is kidney cancer. In one embodiment, the cancer is prostate cancer. In one embodiment, the cancer is esophageal cancer. In one embodiment, the cancer is breast cancer. In one embodiment, the cancer is gastric cancer. In one embodiment, the cancer is kidney cancer. In one embodiment, the cancer is salivary gland cancer. In one embodiment, the cancer is ovarian cancer. In one embodiment, the cancer is uterine carcinoma. In one embodiment, the cancer is lung cancer.

In one embodiment, the FXR agonist is Compound (1) or a pharmaceutically acceptable salt thereof. In another embodiment, the FXR agonist is the sodium salt of compound (1) (ie, compound (1) -Na). In another embodiment, the FXR agonist is an N, N-dietanamine salt of compound (1) (ie, compound (1) -DEA).

In another embodiment, FXR agonist is Compound (2) or a pharmaceutically acceptable salt or amino acid conjugate thereof. In one embodiment, the FXR agonist is a glycine conjugate of compound (2). In one embodiment, the FXR agonist is a taurine conjugate of compound (2). In one embodiment, the FXR agonist is a sarcosine conjugate of compound (2).

The present invention further provides Compound (1) or a pharmaceutically acceptable salt or compound (2) or a pharmaceutically acceptable salt or amino acid conjugate thereof in the manufacture of a medicament for the treatment or prophylaxis of cancer in a subject in need thereof. It relates to the use of.

The invention further relates to compound (1) or a pharmaceutically acceptable salt thereof and compound (2) or a pharmaceutically acceptable salt or amino acid conjugate thereof for use in the treatment or prevention of cancer in a subject in need thereof. .

The invention also provides compounds (1) or their pharmaceutically acceptable salts and compounds (2) or pharmaceutically acceptable salts or amino acid conjugates thereof and pharmaceutically acceptable for treating or preventing cancer in a subject in need thereof. It relates to a pharmaceutical composition comprising an excipient.

The invention further provides for the treatment or prophylaxis of cancer in a subject in need thereof comprising compound (1) or a pharmaceutically acceptable salt thereof and compound (2) or a pharmaceutically acceptable salt or amino acid conjugate thereof Relates to a kit.

1A is a bar graph showing the effect of compound (1) -Na and control diet on the number of liver tumors in multidrug resistant protein 2 (Mdr2 ^{− / −} ) knockout mice. * p <0.01 vs control.
1B is a bar graph showing the effect of Compound (1) -Na and the control diet on the percentage of tumor (greater than 5 mm diameter) reduction in Mdr2 ^{− / −} mice.
2A is a bar graph showing the effect of OCA (Compound (2)) and control diet on the number of Mdr2 ^{− / −} knockout mouse liver tumors.
FIG. 2B is a bar graph showing the effect of OCA and control diet on tumor (more than 5 mm diameter) reduction percentage in Mdr2 ^{− / −} mice.
FIG. 3A is a bar graph showing the effect of Compound (1) -Na and the control diet on the number of liver tumors in Panesoid X receptor (FXR ^{− / −} ) mice.
3B is a bar graph showing the effect of Compound (1) -Na and the control diet on the percentage of tumor (greater than 5 mm diameter) reduction in FXR ^{− / −} mice.
4 is a bar graph showing the effect of Compound (1) -Na and the control diet on the percentage reduction in number of liver tumors in FXR ^{− / −} mice.
FIG. 5A is a bar graph showing the effect of Compound (1) -Na and the control diet on liver / weight ratio reduction percentage in Mdr2 ^{− / −} mice. * p <0.01 vs control.
5B is a bar graph showing the effect of Compound (1) -Na and the control diet on liver / body weight reduction percentage in FXR ^{− / −} mice.
FIG. 6A is a bar graph showing the effect of Compound (1) -Na and the control diet on alanine transaminase (ALT) levels in Mdr2 ^{− / −} and FXR ^{− / −} mice. * p <0.01 vs control.
FIG. 6B is a bar graph showing the effect of Compound (1) -Na and the control diet on aspartate transaminase (AST) levels in Mdr2 ^{− / −} and FXR ^{− / −} mice. * p <0.01 vs control.
FIG. 7A is a bar graph showing the effect of compound (1) -Na and control diet on ileal gene expression of fibroblast growth factor 15 (Fgf15) in Mdr2 ^{− / −} and FXR ^{− / −} mice. * p <0.01 vs control.
FIG. 7B is a bar graph showing the effect of Compound (1) -Na and the control diet on ileal gene expression of small heterodimeric partners (Shp) in Mdr2 ^{− / −} and FXR ^{− / −} mice. * p <0.01 vs control.
FIG. 8 is a bar graph showing the effect of Compound (1) -Na and the control diet on downregulation of cholesterol 7 alpha-hydroxylase (cyp7a1) in Mdr2 ^{− / −} and FXR ^{− / −} mice. * p <0.01 vs control.
9A is a bar graph showing the effect of Compound (1) -Na and control diet on liver gene expression of small heterodimeric partner (Shp) in Mdr2 ^{− / −} and FXR ^{− / −} mice. * p <0.01 vs control.
FIG. 9B is a bar graph showing the effect of Compound (1) -Na and control diet on hepatic gene expression of bile salt transport pump (Bsep) in Mdr2 ^{− / −} and FXR ^{− / −} mice. * p <0.01 vs control.
10A is a bar graph showing the effect of Compound (1) -Na and the control diet on total serum bile acids in Mdr2 ^{− / −} mice. * p <0.01 vs control.
10B is a bar graph showing the effect of Compound (1) -Na3 and the control diet on total serum bile acids in FXR ^{− / −} mice.
11A shows FXR mRNA microarray expression in total tissue of CCA tumors compared to surrounding human tissue.
11B shows FXR mRNA microarray expression in total tissue of CCA tumors sorted according to tumor differentiation grade.
11C shows FXR mRNA expression (qPCR) in CCA tumors compared to normal human liver tissue and surrounding human liver tissue.
11D shows FXR mRNA expression (qPCR) in CCA tumors compared to matched-peripheral human liver tissue.
12A shows TGR mRNA microarray expression in total tissue of CCA tumors compared to surrounding human tissue.
12B shows clinical-pathological parameters: TGR5 mRNA microarray expression in total tissues of CCA tumors at anatomical location and pericardial invasion.
12C shows TGR5 mRNA expression (qPCR) in CCA tumors compared to surrounding human liver tissue.
12D shows TGR5 mRNA expression (qPCR) in CCA tumors compared to matched-peripheral human liver tissue.
13A shows FXR mRNA expression (qPCR) in normal human bile duct cells and CCA cell lines.
13B shows TGR5 mRNA expression (qPCR) in normal human bile duct cells and CCA cell lines.
14A shows representative MRI and liver images of untreated control mice, OCA-treated mice and Compound (4) -treated mice.
14B is a bar graph showing tumor volume fold-change quantified by MRI.
14C shows mRNA expression levels of hepatic ectopic CCA tumor proliferation (ie Ki67 and PCNA ), biliary tract (ie CK19 ) and epithelial (ie ZO-1 ) markers.
14D shows representative immunohistochemical images of hepatic ectopic CCA intratumoral proliferation markers (ie Ki67 and PCNA ) in mice treated with or without OCA or Compound (4).
FIG. 15A shows proliferation and treatment of CCA cells (ie EGI1) treated with 10 or 25 μM OCA for 48 hours as compared to untreated control cells, performed by flow cytometry using CFSE cell proliferation dye staining. MRNA expression levels of proliferation markers are shown.
15B shows representative microscopic images of migration assays and wound-healing assays in CCA cells (ie EGI1), corresponding quantification and representative microscopic images of transwell transfer chambers at 24 hours.
FIG. 15C shows a bar graph of hippocampal oxygen consumption rate (OCR) and metabolic parameters calculated when measuring OCR using a mitochondrial stress test kit in CCA cells (ie EGI1).
15D shows flow cytometry-based apoptosis analysis and pooled data by Annexin V and Propidium Iodide staining in CCA cells treated with or without 10 or 25 μM OCA (ie, EGI1). Representative histograms and corresponding quantifications are shown.
16A shows the proliferation of CCA cells treated with 10 or 25 μM of compound (3) for 48 hours compared to untreated control cells (ie EGI1) and mRNA expression levels of proliferative markers upon treatment with compound (4) .
16B shows transwell migration chambers at 24 hours in migration assays in CCA cells (ie, EGI1), representative microscopic images of wound-healing assays, and corresponding quantified and untreated and Compound (4) -treated cells. Representative microscopic images of and corresponding quantifications are shown.
FIG. 16C shows a bar graph of hippocampal oxygen consumption rate (OCR) and metabolic parameters calculated when measuring OCR using a mitochondrial stress test kit in CCA cells treated with or without Compound 4 (25 μM) (ie, EGI1).
17 shows mRNA expression levels of hepatic ectopic CCA tumor proliferation (ie, Cdc25a, Cyclin D1 and Cyclin D3) markers.

The present invention is based, at least in part, on the discovery that compounds (1) and (2) were effective in treating cancer in animal models predicting cancer. As described in the Examples below, we found that the compounds of the present invention inhibit tumor growth in murine models of spontaneous liver cancer development.

Justice

For convenience, any terms used in the description, examples, and appended claims are collected in this section.

The term "cancer" as used herein refers to any disease characterized by the presence of cancerous tissue in a subject.

As used herein, “cancerous tissue” refers to tissue that includes malignant tumor cells and exhibits abnormal growth and / or hyperproliferative cells of the cells. The cancerous tissue may be a primary malignant tumor resulting from the tissue or organ of origin, or it may be a metastatic malignant tumor that grows in body tissue that is not the source of the tumor of origin.

As used herein, the term "tumor" may include solid tumors or cancers of hematopoietic origin. In some embodiments, the tumor may be characterized by its ability to invade surrounding tissue, metastasize to other parts of the body, and / or its angiogenic activity. Exemplary tumors are produced from hepatocellular carcinoma, gastric cancer, kidney cancer, prostate cancer, adrenal cancer, pancreatic cancer, breast cancer, bladder cancer, salivary gland cancer, ovarian cancer, uterine cancer, and lung cancer.

As used herein, the term “invasive” refers to the process by which cells, cell populations, or malignancies spread from one site to an adjacent site.

As used herein, the term “metastatic” refers to the process by which cells, cell populations, or malignancies spread from one site to a site that is not adjacent to the first site.

As used herein, "hepatocellular carcinoma", "HCC", and "malignant liver cancer" are used interchangeably to refer to primary and secondary (metastatic) tumors derived from liver tissue. The term “refractory hepatocellular carcinoma” as used herein refers to hepatocellular carcinoma that does not respond favorably to antitumor treatment . Thus, "hepatocellular carcinoma that is refractory to treatment" as used herein refers to hepatocellular carcinoma that does not respond favorably or is resistant to treatment, or, alternatively, responds favorably to treatment and then recurs or recurs.

As used herein, the term "cholangiocarcinoma" or "CCA" or "cholangiocarcinoma" or bile duct cancer consists of mutated epithelial cells (or cells characteristic of epithelial differentiation) derived from the bile ducts that drain bile from the liver into the small intestine It is a form of cancer. The proportion of CCA occurs in the cholestatic liver state. Intrahepatic accumulation of bile acids does not directly induce carcinogenesis, but can promote co-carcinogenic effects by promoting bile duct cell proliferation and inflammation and reducing FXR-dependent chemoprotection.

As used herein, the term "compound (1)"

를 지칭하며,

Refers to

It is also known as 6α-ethyl-3α, 7α, 23-trihydroxy-24-nor-5β-cholane-23-hydrogen sulfate. "Compound (1) -Na" or "(1) -Na", also known as 6α-ethyl-3α, 7α, 23-trihydroxy-24-nor-5β-cholan-23-sulfate sodium, is interchangeable And sodium salt of compound (1). Also referred to as "6α-ethyl-3α, 7α, 23-trihydroxy-24-nor-5β-cholane-23-sulfate N, N-diethylethanamine" Known, as used herein, "Compound (1) -DEA" or "1-DEA" are used interchangeably and refer to the N, N-diethylethanamine salt of Compound (1). The structures of compound (1) -Na and compound (1) -DEA are provided below.

.

.

As used herein, "Compound (2)"

를 지칭하며,

Refers to

It is also known as obeticholic acid (OCA), 6-ECDCA, 6-alpha-ethyl kenodeoxycholic acid, or 6α-ethyl-3α, 7α-dihydroxy-5β-cholan-24-oic acid.

As used herein, "Compound (3)"

를 지칭하며,

Refers to

It is also known as 3α, 7α, 11β-trihydroxy-6α-ethyl-5β-cholan-24-oic acid.

As used herein, "Compound (4)"

를 지칭하며,

Refers to

It is also known as 6α-ethyl-23 (S) -methyl-3α, 7α, 12α-trihydroxy-5β-cholan-24-ioic acid. Compound (4) described in US Pat. No. 8,114,862 is a TGR5 modulator. In one of the embodiments, the TGR5 modulator is an agonist.

The term "TGR5 modulator" means any compound that interacts with a TGR5 receptor. The interaction is not limited to compounds that act as antagonists, agonists, partial agonists or inverse agonists of the TGR5 receptor.

In general, the term “agonist” means a compound that enhances the activity of another molecule or receptor site. In the traditional definition, an agonist is a biological agent that binds to a receptor and alters its receptor state, whether or not it is an orthosteric agonist, an allosteric agonist, an inverse agonist, or a co-agonist. Has the action of causing action. In conclusion, agonism is defined as the nature of an agonist or ligand that produces a biological action. More specifically, the TGR5 agonist is a receptor ligand or compound that binds TGR5 and increases the concentration of cyclic adenosine monophosphate (cAMP) by at least 20% in cells expressing the receptor.

As used herein, the phrase “compound of the invention” refers to compound (1), (1) -Na, (1) -DEA, (2), and (3), or a pharmaceutically acceptable salt or amino acid conjugate thereof. Comprehensive

As used herein, the term “treatment” refers to any indication of success in the treatment or alleviation of cancer. Treatment may include, for example, reducing or lessening the severity of one or more symptoms of cancer, or it may include reducing the frequency of symptoms, such as a disease, defect, disorder, or adverse condition that a patient experiences. "Treatment" may refer to a reduction or removal of parts of the body, such as cells, tissues or body fluids, such as blood conditions.

As used herein, the term “prevention” refers to partial or complete prevention of cancer in an individual or population, or in parts of the body, such as cells, tissues, or body fluids (eg, blood). The term “prevention” does not require complete prevention of the disease or condition in the entire treated population of the individual or in cells, tissues or fluids of the individual. The term “treating or preventing”, as used herein, refers to a method for causing some level of treatment or alleviation of cancer and encompassing a range of outcomes for that purpose, including but not limited to total prevention of cancer.

As used herein, the phrase “therapeutically effective amount” is used to induce a tumor to contract and / or to reduce the growth rate of a tumor (eg, to inhibit tumor growth) or to prevent or delay other unwanted cell proliferation in cancer. An effective amount including a sufficient amount is shown. In some embodiments, the effective amount is an amount sufficient to delay the onset of cancer. In some embodiments, the effective amount is an amount sufficient to prevent or delay relapse. An effective amount can be administered in one or more administrations. For HCC or CRC, the effective amount of drug or composition can be as follows: (i) reduce the number of tumor cells; (ii) reduce tumor size; (iii) inhibit, to some extent, hinder, slow down, and preferably stop the infiltration of cancer cells into peripheral organs; (iv) inhibit (ie, slow to some extent and preferably stop) tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay the development and / or recurrence of the tumor, and / or (vii) relieve to some extent one or more symptoms associated with the cancer.

As used herein, the term “therapy” refers to a protocol for dose adjustment and / or timing of the administration of a compound of the invention for the treatment of cancer. Therapy can include active administration and rest periods, as known in the art. Active dosing periods include administration of a compound of the invention over a defined time course, including, for example, the number and timing of doses of the compositions. In some therapies, one or more rest periods during which the compounds are not actively administered may be included, and in certain instances include when the efficacy of such compounds may be minimal.

As used herein, “combination therapy” indicates that a compound of the invention can be administered in conjunction with another therapeutic agent. “With” refers to administration of a compound of the invention as described herein in addition to administration of one treatment mode, such as administration of another therapeutic agent to the same subject, in addition to another treatment mode. As such, “in conjunction with” refers to administration of the first treatment modality before, during, or after the second treatment modality for the subject.

As used herein, "pharmaceutically acceptable" refers to a material that is biologically or otherwise undesirable, such as for example, this material induces any significant undesirable biological effect or interacts in a detrimental manner with other components of the composition in which it is contained. It can be incorporated into a pharmaceutical composition that is administered to a patient without functioning. Pharmaceutically acceptable carriers or excipients are included on inert ingredient guides that meet the required standards of toxicology and manufacturing evaluation and / or are prepared by the US Food and Drug Administration.

As used herein, the term "about" refers to measurable values such as amounts, time periods, etc., as such modifications are appropriate for carrying out the disclosed methods or for preparing and using the disclosed compounds and in the claimed methods. , ± 20% or ± 10% from specified values, ± 5% in some embodiments, ± 1% in some embodiments, and ± 0.1% in some embodiments.

How to treat cancer

The present invention is based, at least in part, on the discovery that the compounds of the present invention are effective in treating cancer in murine models that predict human cancer. Thus, the present application relates to compounds (1), (1) -Na, (1) -DEA, (2), and (3):

Or administering a therapeutically effective amount of an FXR agonist selected from the group consisting of pharmaceutically acceptable salts or amino acid conjugates thereof. In one embodiment, the FXR agonist is compound (1). In one embodiment, the FXR agonist is compound (1) -Na. In another embodiment, FXR agonist is compound (1) -DEA. In one embodiment, the FXR agonist is compound (2). In one embodiment, the FXR agonist is compound (3).

In the methods described herein, exemplary cancers are selected from the group consisting of hepatocellular carcinoma, cholangiocarcinoma, pancreatic cancer, prostate cancer, esophageal cancer, breast cancer, gastric cancer, kidney cancer, salivary gland cancer, ovarian cancer, cervical cancer, bladder cancer, and lung cancer . Appropriate treatment regimens for cancer depend on the type of cells from which the tumor is derived, the stage and severity of the malignancy, and the genetic abnormalities that contribute to the tumor.

Cancer staging systems describe the extent of cancer progression. Generally, a staged system describes how far the cancer has spread and assigns patients to similar prognosis and treatment within the same staged group. In general, the prognosis for tumors that become invasive or metastasized is worse. In one type of timing system, cases are classified into four stages represented by Roman numerals I through IV. In stage I, cancer is often localized and usually curable. Stage II and IIIA cancers are usually more advanced and can invade surrounding tissues and spread to lymph nodes. Stage IV cancers include metastatic cancers that have spread to the outer areas of lymph nodes.

Another staged system is TNM staged, indicating tumor, nodule, and metastasis (Tumor, Nodes, and Metastases) classification. In this system, malignancies are described according to the severity of the individual classification. For example, T classifies the extent of primary tumors from 0 to 4, with 0 representing a malignant tumor having no invasive activity and 4 representing a malignant tumor invading other organs by expansion from the original site. N represents the degree of lymph node involvement, where 0 represents malignant tumor with no lymph node involvement and 4 represents malignant tumor with extensive lymph node involvement. M classifies the extent of metastasis from 0 to 1, where 0 represents malignant tumor without metastasis and 1 represents malignant tumor with metastasis.

These staged systems or variations of these staged systems or other suitable staged systems can be used to describe the tumor. Depending on the stage and characteristics of the cancer, few options are available for the treatment of the cancer. Treatment includes surgery, treatment with sorafenib, and targeted therapies. In general, surgery is first-line treatment for early stage localized cancer. Additional systemic treatments can be used to treat invasive and metastatic tumors.

 According to one aspect of the invention, a method of treating hepatocellular carcinoma (or malignant liver cancer) is provided. In particular, the method comprises treating a subject having a hepatocellular carcinoma in a therapeutically effective amount of a compound of the invention. That is, the present invention relates to the use of a compound of the present invention for the manufacture of a medicament for the treatment of a patient with hepatocellular carcinoma identified or diagnosed as having hepatocellular carcinoma. In a separate embodiment, the method of treatment optionally includes diagnosing or identifying the patient as having hepatocellular carcinoma. The identified patient is then treated with or administered a therapeutically effective amount of a compound of the invention. Hepatocellular carcinoma can be diagnosed in any conventional diagnostic method known in the art including ultrasonography, CT scan, MRI, alpha-fetoprotein evaluation, des-gamma carboxyprothrombin screening, and biopsies.

The invention also provides a method of treating refractory hepatocellular carcinoma comprising treating a patient identified as having a refractory hepatocellular carcinoma in a therapeutically effective amount of a compound of the invention. In certain embodiments, the patient has a sorafenib, regorafenib, anthracycline (e.g. doxorubicin, daunorubicin, epirubicin, idarubicin), a platinum agent (e.g. cisplatin, carboplatin, oxaliplatin , Picoplatin), 5-FU, and capecitabine. Hepatocellular carcinoma that is refractory to a treatment comprising at least one drug selected from the group consisting of. The present invention also relates to refractory hepatocellular carcinoma such as sorafenib, legorafenib, anthracycline (eg doxorubicin, daunorubicin, epirubicin, idarubicin), platinum preparations (eg cisplatin, carboplatin) , Oxaliplatin, picoplatin), 5-FU, and capecitabine, for the use of a compound of the invention for the manufacture of a medicament for the treatment of hepatocellular carcinoma refractory to one or more drugs.

To detect refractory hepatocellular carcinoma, patients undergoing initial treatment can be carefully monitored for signs of resistant, non-responsive or recurrent hepatocellular carcinoma. It consists, for example, of sorafenib, legorafenib, doxorubicin, daunorubicin, epirubicin, idarubicin, cisplatin, carboplatin, oxaliplatin, picoplatin, 5-FU, tegapur and capecitabine. It can be achieved by monitoring the response of the patient's cancer to the initial treatment, which may include one or more drugs selected from the group. Response to the initial treatment, absence of response, or recurrence of cancer can be measured by any suitable method practiced in the art. For example, this can be achieved by evaluation of tumor size and number. Increasing the tumor size, or alternatively the number of tumors, suggests that the tumor is not responding to chemotherapy or that relapse has occurred. The decision is described in Therasse et al, J. Natl. Cancer Inst. 92: 205-216 (2000), in accordance with the "RECIST" standard.

According to another aspect of the invention, a method of preventing or delaying the onset (or hepatocellular carcinoma) of hepatocellular carcinoma, or a method of preventing or delaying the recurrence of hepatocellular carcinoma, is provided, which is prevented or delayed in a prophylactically effective amount of a compound of the invention. Treating the patient in need thereof.

Subjects infected with hepatitis B or hepatitis C or having sclerosis are known to have an increased risk of developing hepatocellular carcinoma. In addition, individuals with acute and chronic hepatic porphyria (acute intermittent porphyria, delayed skin porphyria, congenital coproporphyria, mixed porphyrinosis) and tyrosinemia type I are also at increased risk of developing hepatocellular carcinoma. All of these individuals may be candidates for the methods of the invention for preventing or delaying the onset of hepatocellular carcinoma using a prophylactically effective amount of a compound of the invention. In addition, patients with a family history of hepatocellular carcinoma can also be identified for application of the methods of the invention to prevent or delay the onset of hepatocellular carcinoma. For the purpose of preventing or delaying recurrence of hepatocellular carcinoma, a hepatocellular carcinoma patient treated and in a relaxed or stable or progression-free state is a prophylactically effective amount of a compound of the invention to effectively prevent or delay the recurrence or recurrence of hepatocellular carcinoma Can be treated with

According to one aspect of the invention, a method of treating cholangiocarcinoma (CCA) is provided. In particular, the method comprises treating a subject having a bile duct carcinoma with a therapeutically effective amount of a compound of the invention. That is, the present invention relates to the use of a compound of the present invention for the manufacture of a medicament for the treatment of patients with cholangiocarcinoma identified or diagnosed as having cholangiocarcinoma. In a separate embodiment, the method of treatment optionally includes diagnosing or identifying the patient as having cholangiocarcinoma. The identified patient is then treated with or administered a therapeutically effective amount of a compound of the invention. Cholangiocarcinoma can be diagnosed in any conventional diagnostic method known in the art, including ultrasonography, CT scan, MRI, cancer embryo antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) screening, and biopsies. have.

According to another aspect of the present invention, there is provided a method of preventing or delaying the onset of cholangiocarcinoma, or a method of preventing or delaying the recurrence of cholangiocarcinoma, which patient is in need of a prophylactically effective amount of the compound of the invention It comprises the step of treating.

For the purpose of preventing or delaying the recurrence of cholangiocarcinoma, a prophylactically effective amount of a compound of the present invention to effectively prevent or delay the recurrence or recurrence of cholangiocarcinoma in a treated, relieved, stable or progression-free state Can be treated with

According to another aspect of the present invention, a method of inhibiting proliferation and migration of CCA cells by FXR agonists is provided.

According to another aspect of the invention, a method of inhibiting proliferation and migration of CCA cells by a therapeutically effective amount of a compound of the invention is provided. According to another aspect of the invention, there is provided a method of inhibiting the progression of CCA, comprising treating a patient in need of treatment with a therapeutically effective amount of a compound of the invention.

One embodiment is a method of treating pancreatic cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating prostate cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating kidney cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating prostate cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating esophageal cancer by administering a therapeutically effective amount of a compound of the present invention. Another embodiment is a method of treating breast cancer by administering a therapeutically effective amount of a compound of the invention. One embodiment is a method of treating gastric cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating kidney cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating salivary gland cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating ovarian cancer by administering a therapeutically effective amount of a compound of the invention. One embodiment is a method of treating cervical cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating bladder cancer by administering a therapeutically effective amount of a compound of the invention. Another embodiment is a method of treating lung cancer by administering a therapeutically effective amount of a compound of the invention.

If a compound of the invention is useful for the treatment of a cancer described herein, such compound may be co-administered with a second agent. Second agents useful in the methods of treating cancer provided herein include any known class of anticancer agents, such as radiation therapy, surgery, alkylating agents, antimetabolic agents, anthracyclines, campotesins, vinca alkaloids, taxanes or platinum, Other anti-tumor agents known in the art. Such anticancer and antitumor agent classifications are known in the art and used according to their simple and general meaning.

Exemplary anticancer agents include, but are not limited to: ABRAXANE; Abiraterone; Ace-11; Aclarubicin; Ashibisin; Acodazole hydrochloride; Acronin; Actinomycin; Acylpulbene; Adesiphenol; Adozelesin; Adriamycin; Aldesleukin; All trans-retinoic acid (ATRA); Altretamine; Ambamustine; Ambomycin; Amethanetron acetate; Amidox; Amifostine; Aminoglutetimide; Aminolevulinic acid; Amrubicin; Amsacrine; Anagrelide; Anastrozole; Andrographolide; Antarelix; Anthracycin; Apidicholine glycinate; Apurinic acid; Ara-CDP-DL-PTBA; Arginine deaminase; ARRY-162; ARRY-300; ARRY-142266; AS703026; Asparaginase; Asperin; Asulacrine; Atamestan; Atrimustine; Avastin (AVASTIN); Axinastatin 1; Axinastatin 2; Axinastatin 3; Azasetron; Azatoxins; Azatyrosine; Azacytidine; AZD8330; Azethepa; Azotomycin; Balanol; Batimastad; BAY 11-7082; BAY 43-9006; BAY 869766; Bendamustine; Benzochlorine; Benzodepa; Benzoylstaurosporin; Beta-aletin; Betaclomycin B; Betulinic acid; b-FGF inhibitors; Bicalutamide; Bisantrene; Bisaziridinylspermine; Bisnaphide; Bisnapid dimesylate; Bistratene A; Bisantrene hydrochloride; Bleomycin; Bleomycin sulfate; Busulfan; Bizelesin; Breplate; Bortezomib; Brequinar sodium; Bropyrimin; Budotitanium; Butionine sulfoximine; Bryostatin; Cocktinomycin; Calusosterone; Calcipotriol; Calfostine C; Camptothecin derivatives; Capecitabine; Carboxamide-amino-triazole; Carboxyxamidotriazoles; CaRest M3; CARN 700; Carracemide; Carbetatimers; Carboplatin; Carmustine; Carrubicin hydrochloride; Kazelesin; Castanospermine; Secropin B; Cedefingol; Celecoxib; Setlorellix; Chlorins; Chloroquinoxaline sulfonamides; Cicafrost; Chlorambucil; Chlorofucin; Sirolemycin; Cisplatin; CI-1040; Cis-porphyrin; Cladribine; Clomiphene analogs; Clotrimazole; Cholismycin A; Cholismycin B; Combretastatin A4; Combretastatin analogs; Congeninin; Crampescidine 816; Crisnatol; Crisnatol mesylate; Cryptophycin 8; Cryptophycin A derivatives; Curacin A; Cyclopentanetraquinone; Cycloplatam; Cifemycin; Cyclophosphamide; Cytarabine; Cytarabine oxphosphate; Cytolytic factor; Cytostatin; Dacarbazine; Dactinomycin; Daunorubicin; Daunorubicin hydrochloride; Decarbazine; Daclicksimab; Dasatinib; Decitabine; Dehydrodidemnin B; Deslorelin; Dexamethasone; Dexiphosphamide; Dexlaonic acid; Dexverapamil; Dexormaplatin; Dezaguanine; Dezaguanine mesylate; Diajikuon; Didemnin B; Dedox; Diethylnorspermine; Dihydro 5 azacytidine; Dihydrotaxol; 9-dioxamycin; Diphenyl spiromostin; Docosanol; Dolacetron; Docetaxel; Doxorubicin; Doxorubicin hydrochloride; Doxyfluidine; Droloxifene; Droroxifene citrate; Dromostanolone propionate; Dronabinol; Duazomycin; Duocarmycin SA; Epselen; Echomustine; Edelfosine; Edrecolomab; Edda trexate; Eplonitine hydrochloride; Eflornithine; Element; Emitepur; Elsammitrusin; Enroplatin; Enpromate; Epipropidine; Epirubicin; Epirubicin hydrochloride; Epristeride; Erbulosol; Eribulin; Esorubicin hydrochloride; Esturamustine; Esthramustine phosphate sodium; Ethanidazol; Etoposide; Etoposide phosphate; Etorine; Exemestane; Fadrosol; Fadrosol hydrochloride; Pazarabine; Fenretinide; Filgrastim; Finasteride; Flavopyridols; Flezelastine; Fluasterone; Phloxuridine; Fludarabine phosphate; Fludarabine; Fluorodaunorubicin hydrochloride; Porfenix; Formemstan; Fluorouracil; Phloxuridine; Flurocitabine; Phosquidone; Postriecin sodium; Postriecin; Potemustine; Gadolinium texaphyrin; Gallium nitride; Gallocitabine; Ganellilix; Gelatinase inhibitors; Gemcitabine; Zeldanamycin; Gosipol; GDC-0973; GSK1120212 / trametinib; Herceptin; Hydroxyurea; Hepsulpam; Heregulin; Hexamethylene bisacetamide; Hypericin; Ibandronic acid; Ibrutinib; Idarubicin; Idarubicin hydrochloride; Ifosfamide; Canphosphamide; Monomorphine; Ifoplatin; Idoxifen; Isdramantone; Monomorphine; Ilomatstat; Imidazoacridone; Imatinib (eg GLEEVEC); Imiquimod; Iniparib (BSI 201); Iobenguan; Iododoxorubicin; Ifomeranol; Irinotecan; Irinotecan hydrochloride; Irsogladine; Isobenazole; Iso homohalicondrin B; Itacrone; Imimofosine; Interleukin IL-2 (including recombinant interleukin II; or rlL.sub.2); Interferon alfa-2a; Interferon alpha-2b; Interferon alpha-n1; Interferon alpha-n3; Interferon beta-1a; Interferon gamma-1b; Jasplatinolide; Kahalarid F; Lamelalin N triacetate; Lanreotide; Reinamycin; Renograstim; Lentinan sulfate; Leptolstatin; Letrozole; Leuprorelin; Levamisol; Lenalidomide; Levatinib; Liarosol; Lissocleanamide 7; Lovaplatin; Rombrisin; Rometrexole; Rodidamine; Loxoxanthrone; Lovastatin; Roxoribin; Lutetocan; Lutetium texaphyrin; Lysophylline; Lanleotide acetate; Lapatinib; Letrozole; Leucovorin; Leuprolide acetate; Liarosol hydrochloride; Rometrexole sodium; Romustine; Loxoxanthrone hydrochloride; Pomalidomide; LY294002; Maytansine; Mannostatin A; Marimastat; Masoprocol; Maspin; Matrilysine inhibitors; Menogaryl; Merbaron; Meterelin; Methionase; Metoclopramide; MIF inhibitors; Mifepristone; Miltefosine; Myrimos team; Mitoguazone; Mitolactol; Mitona feed; Mitoxantrone; Mofarotene; Molgramos team; Fur mall; Mycaperoxide B; Myriaporon; Maytansine; Mechlorethamine hydrochloride; Megestrol acetate; Melengestrol acetate; Melphalan; Mercaptopurine; Methotrexate; Methotrexate sodium; Metoprin; Metaturdepa; Mitindomide; Mitocasin; Mitochromen; Mitogiline; Mitomalcin; Mitomycin; Mitosper; Mitothene; Mitoxantrone hydrochloride; Mycophenolic acid; Naparelin; Nagre tip; Napabin; Naphterpins; Nartograstim; Nedaplatin; Nemorubicin; Neridronic acid; Nilutamide; Nisamycin; Nitric oxide modulators; Nitroxide antioxidants; Nitrile; Nocodazole; Nogalamycin; Oblimersen (GENASENSE); Octreotide; Ocisenone; Olaparib (LYNPARZA); Oligonucleotides; Onapristone; Ondansetron; Oracin; Oral cytokine inducers; Ormaplatin; OxySuran; Oxaloplatin; Ostherone; Oxaliplatin; Oxaunomycin; Palauamine; Palmitolylicin; Pamidronic acid; Panaxitriol; Panomifen; Parabactin; PARP (polyADP ribose polymerase) inhibitors; Pazelliptin; Pegaspargase; Peldesin; Pentosan polysulfate sodium; Pentostatin; Pentrozole; Perflubrones; Perphosphamide; Peryl alcohol; Phenazinomycin; Phenyl acetate; Phosphatase inhibitors; Pichibanyl; Pilocarpine hydrochloride; Pyrarubicin; Pyritrexime; Placetin A; Placetin B; Porphyromycin; Prednisone; Prostaglandin J2; Pyrazoloacridine; Paclitaxel; PD035901; PD184352; PD318026; PD98059; Peliomycin; Pentamustine; Petromycin sulfate; PKC412; Fibobroman; Capfosulfan; Pyroxanthrone hydrochloride; Plicamycin; Plomethane; Grape phytotoxin; Polyphenol E; Porphymer sodium; Porphyromycin; Prednismustine; Procarbazine; Procarbazine hydrochloride; Puromycin; Puromycin hydrochloride; Pyrazopurin; Raltitrexide; Lamosetron; Retelliptin demethylation; Lysine; Rituximab; RII retinamide; Rogletimide; Lohitukine; Romultide; Loquinimex; Rubizinone B1; Luboksil; Ribophrine; Romidepsin; Lucaparib; Safingol; Sapgol hydrochloride; Sinetopin; Sarcopitol A; Sargramos team; Semustine; Sizopyran; Small aliphatic acid; Sodium borocaptate; Sodium phenylacetate; Solberol; Sonermin; Sorafenib; Sunitinib; Spapoic acid; Spicamycin D; Spiromostin; Splenopenpentin; Spongystatin 1; Spongystatin 2; Spongystatin 3; Spongystatin 4; Spongystatin 5; Spongystatin 6; Spongystatin 7; Spongystatin 8; And spongystatin 9; Squalane; Styphiamide; Stromelysin inhibitors; Sulfinosine; Suradista; Suramin; Swainsonine; SB239063; Cellumetinib / AZD6244; Simtragen; SP600125; Sparfosate sodium; Spartomycin; Spirogermanium hydrochloride; Spiroplatin; Streptonigrin; Streptozosin; Sulofenur; Thalimustine; Tamoxifen methiodide; Thalazoparib (BMN 673); Tauromustine; Tazarotene; Tecogallan sodium; Tegapur; Tellulapyryllium; Temophorpine; Temozolomide; Teniposide; Tetrachlorodecaoxide; Tetrazomine; Thaliblastine; Thiocoralin; Thrombopoietin; Thymalfasin; Thymopoietin receptor agonists; Thymotrinan; Tyrapazamine; Titanocene bichloride; Topsentin; Toremifene; Tretinoin; Triacetyluridine; Trisiribin; Trimetrexate; Tryptorelin; Trophysetron; Turosteride; Tyrphostin; Thalisomycin; TAK-733; Taxotere; Tegapur; Teloxtron hydrochloride; Theroxylone; Testosterone; Thiamiprine; Thioguanine; Thiotepa; Thiazopurin; Tyrapazamine; Toremifene citrate; Trastuzumab; Trestolone acetate; Trisiribin phosphate; Trimetrexate; Trimetrexate glucuronate; Tryptorelin; Tubolosol hydrochloride; Tumor necrosis factor-associated apoptosis-inducing ligand (TRAIL); UBC inhibitors; Ubenimex; U0126; Uracil mustard; Uredepa; Vapreotide; Variolin B; Belaresol; Bellifarib (ABT-888); Veramine; Berteporphine; Vinorelbine; Vinsaltin; Nontaxin; Vinblastine; Vinblastine sulfate; Vincristine sulfate; Bindesin; Vindesine sulfate; Vinepidine sulfate; Vin glycinate sulfate; Vinurosin sulphate; Vinorelbine tartrate; Binrocidine sulfate; Vinzolidine sulfate; Borosol; Wortmannin; XL518; Zanotone; Geniplatin; Zillaskov; Ginostatin stymalamer; Ginostatin; And zorubicin hydrochloride.

Other exemplary anticancer agents include erbulosol (eg, R-55104); Dolastatin 10 (eg, DLS-10 and NSC-376128); Mibobulin isethionate (eg, CI-980); NSC-639829; Discothemoldes (eg, NVP-XX-A-296); ABT-751 (Abbott; for example E-7010); Altorhirtin A; Altorhirtin C; Semadothine hydrochloride (eg, LU-103793 and NSC-D-669356); CEP 9722; Epothilone A; Epothilone B; Epothilone C; Epothilone D; Epothilone E; Epothilone F; Epothilone B N-oxide; Epothilone A N-oxide; 16-aza-epothilone B; 21-aminoepothilone B; 21-hydroxyepothilone D; 26-fluoroepothilone; Auristatin PE (eg, NSC-654663); Soblidotin (eg, TZT-1027); LS-4559-P from Pharmacia (eg, LS-4577); LS-4578 (Pharmacia; for example LS-477-P); LS-4477 from Pharmacia; LS-4559 from Pharmacia; RPR-112378 from Aventis; DZ-3358 from Daiichi; FR-182877 (Fujisawa; eg, WS-9265B); GS-164 from Takeda; GS-198 from Takeda; Hungarian Academy of Sciences (KAR-2); BSF-223651 (BASF; for example ILX-651 and LU-223651); SAH-49960 from Lilly / Novartis; SDZ-268970 from Lilly / Novartis; AM-97 from Armad / Kyowa Hakko; AM-132 (Armad); AM-138 from Armad / Kyowa Hakko; IDN-5005 from Indena; Cryptophycin 52 (eg, LY-355703); AC-7739 (Ajinomoto; eg, AVE-8063A and CS-39.HCl); AC-7700 (Ajinomoto; eg, AVE-8062; AVE-8062A; CS-39-L-Ser.HCl; and RPR-258062A); Bitilibuamide; Tubulinin A; Canadensole; CA-170 from Curis, Inc .; Centauleidine (eg, NSC-106969); T-138067 (Tularik; for example T-67; TL-138067 and TI-138067); COBRA-1 from Parker Hughes Institute (eg, DDE-261 and WHI-261); Kans State University (H10); Kansas State University (H16); Oncosidine A1 (eg, BTO-956 and DIME); DDE-313 from Parker Hughes Institute; Fizianolide B; Laulimide; Parker Hughes Institute (SPA-2); Parker Hughes Institute (eg SPIKET-P); 3-IAABU (Cytoskeleton / Mt. Sinai School of Medicine; for example MF-569); Narcosin (eg, NSC-5366); Nascarpin; D-24851 from Asta Medica; A-105972 from Abbott; Hemiasterlin; 3-BAABU (Cytoskeleton / Mt. Sinai School of Medicine; for example MF-191); Arizona State University (TMPN); Vanadocene acetylacetonate; T-138026 from Tularik; Monsartrol; Inanosine (eg, NSC-698666); 3-IAABE (Cytoskeleton / Mt. Sinai School of Medicine); A-204197 from Abbott; T-607 (Tularik; for example T-900607); RPR-115781 from Aventis; Leuterorobins (eg, desmethyl leuterobin; desaethyl leuterobin; isoleuterobin A; and Z-leuterobin); Carbaioside; Caribaolin; Haliconerin B; D-64131 from Asta Medica; D-68144 from Asta Medica; Diazonamide A; A-293620 from Abbott; NPI-2350 (Nereus); Tacalonolide A; TUB-245 from Aventis; A-259754 from Abbott; Diozostatin; (-)-Phenyllahistin (eg, NSCL-96F037); D-62638 from Asta Medica; D-62636 from Asta Medica; Myoseberine B; D-43411 (Zentaris; for example D-81862); A-289099 from Abbott; A-318315 from Abbott; HTI-286 (eg SPA-110; trifluoro acetate salt) (Wyeth); D-82317 from Zentaris; D-82318 from Zentaris; SC-12983 (NCI); Resverastatin phosphate sodium; BPR-OY-007 from National Health Research Institutes; And SSR-250411 (Sanofi)); Goserelin; Leuprolide; Trytolide; Homoharingtonin; Topotecan; Itraconazole; Deoxyadenosine; Sertraline; Pitavastatin; Clofazimin; 5- nonyloxytryptamine; Bemurafenib; Dabrafenib; Gefitinib (IRESSA); Erlotinib (TARCEVA); Cetuximab (ERBITUX); Lapatinib (TYKERB); Panitumumab (VECTIBIX); Vandetanib (CAPRELSA); Afatinib / BIBW2992; CI-1033 / canertinib; Neratinib / HKI-272; CP-724714; TAK-285; AST-1306; ARRY334543; ARRY-380; AG-1478; Dacomitinib / PF299804; OSI-420 / Desmethyl Erlotinib; AZD8931; AEE726; Pelitinib / EKB-569; CUDC-101; WZ8040; WZ4002; WZ3146; AG-490; XL647; PD153035; 5- azathioprine; 5-aza-2'-deoxycytidine; 17-N-allylamino-17-demethoxygeldanamycin (17-AAG); 20-epi-1,25 dihydroxyvitamin D3; 5 ethynyluracil; And BMS-599626.

One embodiment is a method of treating a patient with hepatocellular carcinoma (optionally refractory) by administration of a compound of the invention in combination with capecitabine and / or PLX4032 (Plexxikon).

Another embodiment is a method of treating hepatocellular carcinoma (optionally refractory) by administration of a compound of the invention in combination with capecitabine, geloda, and / or CPT-11.

Another embodiment is a method of treating a patient with hepatocellular carcinoma (optionally refractory) or a patient with unresectable or metastatic hepatocellular carcinoma by administration of a compound of the invention in combination with capecitabine and irinotecan.

Another embodiment is a method of treating a patient with non-resectable or metastatic hepatocellular carcinoma by administration of a compound of the invention in combination with interferon alpha or capecitabine.

Another embodiment is a method of treating a patient with pancreatic cancer by administering a compound of the invention in combination with gemcitabine.

One of the embodiments is a method of treating a patient with CCA by administering another agent or agents or a compound of the present invention in combination with them.

One of the embodiments is a method of treating a patient with CCA by administering gemcitabine, cisplatin or a compound of the invention in combination thereof.

Pharmaceutical composition

A "pharmaceutical composition" is a formulation containing a compound of the invention in a form suitable for administration to a subject. In one embodiment, the pharmaceutical composition is that of a large amount or in unit dosage form. It may be advantageous to formulate the composition in unit dosage form for ease of administration and uniformity of the formulation. Specifications for dosage unit forms are dictated and directly depend on the inherent characteristics of the active reagents and the particular therapeutic effect to be achieved.

Possible formulations include those suitable for topical use, including oral, sublingual, buccal, parenteral (eg, subcutaneous, intramuscular, or intravenous), rectal, transdermal, intranasal and inhaled administration. The most suitable means of administration for a particular patient will depend on the nature and severity of the disease being treated or on the nature of the active compound and the nature of the active compound, but where possible oral administration can be used for the prevention and treatment of cancer. Formulations suitable for oral administration may be presented as discrete units, each containing a predetermined amount of active compound, such as tablets, capsules, cachets, lozenges; As a powder or granules; As a solution or a suspension in an aqueous or non-aqueous liquid; Or as oil-in-water or water-in-oil emulsions.

Formulations suitable for sublingual or buccal administration include compounds of the present invention and pastilles comprising the active compounds, typically in lozenges comprising flavor bases such as sugar and acacia or tragacanth and inert bases such as gelatin and glycerin or sucrose acacia. (pastille).

Formulations suitable for parenteral administration typically include sterile aqueous solutions containing the active compound at a predetermined concentration; The solution may be isotonic with the blood of the intended recipient. Additional formulations suitable for parenteral administration include formulations containing physiologically appropriate co-solvents and / or complexing agents such as surfactants and cyclodextrins. Oil-in-water emulsions are also suitable formulations for parenteral formulations. Although such solutions may be administered intravenously, they may also be administered by subcutaneous or intramuscular injection.

Formulations suitable for rectal administration may be presented as unit-dose suppositories comprising the compounds of the invention in one or more solid carriers, such as cocoa butter, that form a suppository base.

Formulations suitable for topical or intranasal application include ointments, creams, lotions, pastes, gels, sprays, aerosols and oils. Suitable carriers for such formulations include petrolatum, lanolin, polyethylene glycols, alcohols, and combinations thereof.

Formulations of the invention are typically prepared by mixing the compounds of the invention with a liquid or finely divided solid carrier or both in a uniform and intimately necessary proportion, and then, if necessary, by molding the resulting mixture into the desired shape, It may be prepared by any suitable method.

For example, tablets may be prepared by compressing an intimate mixture comprising the active ingredient and one or more optional ingredients such as binders, lubricants, inert diluents, or powders or granules of surfactant dispersants, or with powdered active ingredients. It can be prepared by molding an intimate mixture of inert liquid diluents. Suitable formulations for administration by inhalation include fine particle dusts or sprays that can be produced by various types of metered doses of pressurized aerosols, nebulizers, or pulverizers.

For pulmonary administration via the oral cavity, the particle size of the powder or droplets is typically in the range of about 0.5 to 10 μm, or 1 to 5 μm, to ensure delivery to the bronchial aqueous phase. For nasal administration, particle sizes in the range of about 10 to 500 μm can be used to ensure intranasal retention.

Metered dose inhalers are typically pressurized aerosol dispensers containing a suspension or solution formulation of a compound of the invention in a liquefied propellant. In use, these instruments typically eject the formulation through a valve adjusted to deliver a metered dose of about 10 to 150 μm, producing a fine particle spray containing the active ingredient. Suitable propellants include certain chlorofluorocarbon compounds such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane and mixtures thereof. The formulation may further contain one or more co-solvents such as ethanol surfactants such as oleic acid or sorbitan trioleate, antioxidants and suitable flavoring agents.

Nebulizers are commercially available instruments that convert solutions or suspensions of the active ingredient into therapeutic aerosol sprays by acceleration through narrow venturi holes of compressed gas, typically air or oxygen, or by ultrasonic agitation. Formulations suitable for use in nebulizers consist of the active ingredient in a liquid carrier and contain up to about 40% w / w of formulation and may comprise less than about 20% w / w. The carrier is preferably water or an aqueous diluted alcohol solution, preferably made isotonic with the body fluid, for example by the addition of sodium chloride. Optional additives include preservatives such as methyl hydroxy-benzoate, antioxidants, flavors, volatile oils, buffers and surfactants when the formulation is not prepared sterile.

Formulations suitable for administration by inhalation include finely ground powders that can be delivered by pulverulent or put into the nasal passages in a nasal manner. In the mill, the powder is contained in a capsule or cartridge, typically made of gelatin or plastic, which is delivered by air, which is perforated or opened in use and entered through the instrument upon inhalation, or by a manually operated pump. The powder used for the pulverization consists of the active ingredient alone or of a mixed powder comprising the active ingredient, an appropriate powder diluent such as lactose, and an optional surfactant. The active ingredient typically comprises about 0.1-100% w / w of the formulation.

In a further embodiment, the present invention provides a pharmaceutical composition comprising, as an active ingredient, a compound of the present invention with, and / or a mixture thereof with at least one pharmaceutical carrier or diluent.

The carrier is pharmaceutically acceptable and must be compatible with the other ingredients in the composition, i.e. without adversely affecting the other ingredients. The carrier may be solid or liquid and is preferably formulated in a unit dose formulation, for example a tablet, which may contain from about 0.05 to 95% by weight of active ingredient. If desired, other physiologically active ingredients may also be incorporated into the pharmaceutical compositions of the present invention.

In addition to the components specifically mentioned above, the formulations of the present invention may include other substances known to those skilled in the pharmaceutical art with respect to the type of formulation in question. For example, formulations suitable for oral administration may include flavoring agents and formulations suitable for intranasal administration may include perfumes.

In one embodiment, the pharmaceutical composition comprises about 0.1-1500 mg, 0.2-1200 mg, 0.3-1000 mg, 0.4-800 mg, 0.5-600 mg, 0.6-500 mg, 0.7-400 mg, 0.8-300 mg, Dosage forms comprising a compound of the invention in a daily total amount of 1 to 200 mg, 1 to 100 mg, 1 to 50 mg, 1 to 30 mg, 4 to 26 mg, 5 to 25 mg, or 5 to 10 mg. Is administered.

The compounds of the present invention are combined with other hepatocellular carcinoma therapeutic drugs such as anticancer chemotherapy drugs, hormones, biological response modifier (s), and other angiogenesis inhibitors; Or in combination with immunotherapy or gene therapy.

Example 1.Synthesis of Compound (1)

Compound (1) may be prepared by methods known in the art (eg, those known in U.S. Patent No. 7,932,244). For example, compound (1) can be prepared by a process as shown in Scheme 1 and disclosed in WO 2014/066819.

Scheme 1

Step 1 is esterification of compound (2) to obtain compound (4). Step 2 is a reaction for forming compound (5) from compound (4). Step 3 is the protection of the hydroxyl group at the C-3 position of compound (5) to yield compound (6). Step 4 is an oxidative cleavage of compound (6) to yield compound (7). Step 5 is reduction of compound (7) to yield compound (8). Step 6 is sulfonation of compound (8) to yield the sodium salt (1-Na) of compound (1). The sodium salt of compound (1) may be in its free acid form (ie, compound (1)) or other salt form (eg, compound (1) -DEA or N, N of compound (1) according to procedures known in the art). Diethanamine salt).

Example 2. Synthesis of Compound (2)

Compound (2) is prepared by conventional methods (e.g., those described in US Publication No. 2009/0062526, US Patent No. 7,138,390, and WO 2006/122977), such as Compound (1) (O) as shown in Scheme 2 below. Beticicolic acid, or OCA) can be prepared by the following six-step synthesis.

Scheme 2

The process is a six-step synthesis. Step 1 is the esterification of C-24 carboxylic acid of 7-keto lithocholic acid (KLCA) using methanol and heat in the presence of an acidic catalyst to prepare methyl ester compound (7). Step 2 is the formation of silyleneol ether from the compound using a strong base to prepare compound (8) followed by treatment with chlorosilanes. Step 3 is an aldol condensation reaction of the silyleneol ether of compound (8) and acetaldehyde for preparing compound (10). Step 4 is saponification of the C-24 methyl ester of compound (10) to make compound (11). Step 5 is hydrogenation of the 6-ethylidene moiety of compound (11) to make compound (12). Step 6 is the selective reduction of the 7-keto group to 7a-hydroxy group of compound (12) for preparing compound (1).

Example 3. Mdr2 ^-/-  And FXR ^-/-  Liver Cancer Formation in Mice

Multidrug resistance protein 2 (Abcb4) is a member of the ATP-binding cassette (ABC) transporter superfamily. Multidrug resistant protein 2 gene knockout mice (Mdr2 ^{− / −} ) provide an in vivo model of natural liver cancer formation (Katzenellengoben, et al. Mol. Cancer Res. 2007, 5, 11, 1159-1170). Mice lacking the Abc4 protein encoded by the multidrug resistance-2 gene develop periinflammatory inflammation and cholestatic liver disease resulting in the development of hepatocellular carcinoma.

Panesoid X receptor protein (FXR) is a nuclear receptor that acts as a bile acid sensor that controls bile acid homeostasis. The receptor is highly expressed in the liver and other organs. FXR knockout (FXR ^-/- ) mice develop hepatocellular adenocarcinoma and carcinoma after 15 months of age (Yang, et al. Cancer Res., 2007, 67, 863).

The effect of OCA, Compound (1) -Na, and a control diet in Mdr2 ^{− / −} and FXR ^{− / −} mice on the development of hepatocellular carcinoma was evaluated. OCA is an FXR agonist, while Compound (1) -Na is a dual FXR / TGR5 agonist. The titer of Compound (1) -Na over FXR is about 10 times greater than OCA.

Study design

Mdr2 ^{− / −} and FXR ^{− / −} mice were randomly divided into three experimental groups. Mice received either a specific rodent diet or a diet supplemented with OCA, Compound (1) -Na, or control diet for 15 months. All mice were housed under pathogen free conditions and allowed to drink water freely in a temperature-controlled room (23 ° C.) at 12-hour light / dark cycles. The Bari University Ethics Committee has approved the experimental set-up, which has also been certified by the Italian Ministry of Health according to internationally accepted guidelines for animal care. After 16 months, mice were sacrificed and serum, liver and intestines collected. The total number of liver tumors was counted and the diameter of each tumor was measured.

Treatment group

Group 1: Control Diet

Mice (n = 4) received a control diet for 15 months.

Group 2: OCA

Mice (n = 8) received a control diet supplemented with OCA at a dose of 10 mg / kg for 15 months.

Group 3: compound (1) -Na

Mice (n = 15) received a control diet supplemented with Compound (1) -Na at a dose of 10 mg / kg for 15 months.

result

Hepatic inflammation and toxicity induced by bile salts in Mdr2 ^{− / −} mice results in the development of hepatocellular dysplasia. At 16 months of age, liver tumors developed in nearly 100% of Mdr2 ^{− / −} control mice. Spontaneous hepatocellular carcinoma occurred in 16-month-old FXR ^-/- mice.

Tumor Reduction and Size

1A and 2A show the effect of compound (1) -Na, OCA, and the control on reducing the number of tumors in Mdr2 ^{− / −} mice. Compound (1) -Na clearly prevented the development of hepatocellular carcinoma compared to the control in this study. Little statistical significance was observed in the Compound (1) -Na group (p = 0.055) but was not achieved due to the small number (n = 2) of control animals used in the study. Mdr2 ^{− / −} control mice and OCA treated mice showed clearly identifiable liver tumors, whereas small tumors were identified in the Compound (1) -Na group. 1B and 2B show the effect of compound (1) -Na, OCA, and control on the percentage reduction of tumors with diameters greater than 5 mm. Almost 80% of tumors identified in OCA and controls had diameters greater than 5 mm in Mdr2 ^{− / −} mice. In contrast, FXR ^{− / −} mice treated with Compound (1) -Na, OCA, and controls exhibited some large liver tumors suggesting that the prevention of hepatocellular carcinoma development is mostly FXR dependent (FIGS. 3A, 3B and 4).

Liver weight / weight (LW / BW)

5A and 5B describe the effect of Compound (1) -Na and the control on the percentage ratio of liver weight and body weight. Consistent with previously generated data, Compound (1) -Na treated Mdr2 ^{− / −} mice showed a significant reduction in LW / BW ratio compared to control and OCA treated Mdr2 ^{− / −} mice. No difference in LW / BW ratio was observed in the FXR ^-/- group.

Biochemical parameters

Compound (1) -Na and controls for levels of liver enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST) to assess liver damage in Mdr2 ^-/- and FXR ^-/- mice The effect of was analyzed. As shown in FIGS. 6A and 6B, treatment with Compound (1) -Na significantly reduced ALT and AST levels in Mdr2 ^{− / −} mice. However, no differences were observed in ALT and AST levels in FXR ^{− / −} treated mice.

Presidential FXR Target Gene Expression

To demonstrate the involvement of FXR in the prevention of hepatocellular carcinoma, the effects of Compound (1) -Na and control on ileal FXR target gene expression were evaluated. As expected, both OCA and Compound (1) -Na stimulated fibroblast growth factor 15 (Fgf15) and small heterodimeric partner (Shp) gene expression only in the Mdr2 ^{− / −} group (FIGS. 7A and 7B). .

Liver FXR Target Gene Expression

Cholesterol 7 alpha-hydroxylase (cyp7a1) is a rate determining enzyme in the traditional biosynthetic pathway that converts cholesterol into bile acids. Compound (1) -Na and OCA both inhibited Cyp7a1 gene expression only in Mdr2 ^{− / −} mice (FIG. 8). Bile salt transport pump (Bsep) is a membrane protein that uses ATP hydrolysis energy to actively transport bile salts. As shown in FIGS. 9A and 9B, Compound (1) -Na administration induced hepatic Bsep activation in Mdr2 ^{− / −} mice. OCA did not promote hepatic Bsep induction, suggesting that OCA is unlikely to have hepatic activity in Mdr2 ^{− / −} mice, in contrast to Compound (1) -Na, which efficiently activates FXR in the gut and liver.

Serum total bile acid

Compound (1) -Na significantly reduced serum bile acid levels in Mdr2 ^{− / −} mice (FIG. 10A). FXR dependence of this finding was confirmed by the observation that no reduction occurred in FXR ^{− / −} mice (FIG. 10B).

Example 4 Differential Effects of FXR or TGR5 Activation in Cholangiocarcinoma Progression

Way

FXR and TGR5 expression is different in human CCA compared to normal human bile duct cells (NHC) as well as in CCA human biopsies and controls using two approaches (mRNA microarrays and qPCR) and two different patient cohorts (Copenhagen and San Sebastian). It was measured (by qPCR) in the cell line. Growth of ectopic models of human CCA in immunodeficient mice was assessed by magnetic resonance imaging (MRI) under chronic administration of specific FXR or TGR5 agonists (OCA or Compound 4, respectively, 0.03% in Chow for 2 months; Intercept Pharmaceuticals). . The differential effects of FXR or TGR5 activation were also evaluated for proliferation, migration and mitochondrial energy metabolism in CCA and NHC cells in vitro .

result

FXR is downregulated and TGR5 is upregulated in human CCA tissue samples compared to normal peripheral liver tissue and normal hepatic bile duct in the Copenhagen and San Sebastian cohorts by mRNA microarray and qPCR, respectively. TGR5 can promote CCA progression through regulation of proliferation, migration and mitochondrial energy metabolism, while inhibiting FXR activation. Modulation of FXR and TGR5 activity represents a potential therapeutic strategy for CCA. In vitro, FXR was found to be downregulated in different human CCA cell lines compared to NHCs and TGR5 is upregulated.

As shown in FIGS. 11A-11D, FXR expression is reduced in CCA tumors and correlates with tumor differentiation: FIG. 11A shows the total tissue (n = 104) of CCA tumors compared to surrounding human tissue (n = 60). FXR mRNA microarray expression in (Copenhagen cohort) (Man-Whitney test); 11B shows FXR mRNA microarray expression in total tissue of CCA tumors sorted according to tumor differentiation grade: well-differentiated (n = 10), moderately-differentiated (n = 34) or hardly-differentiated (n = 9) (Copenhagen cohort) (Man-Whitney test); 11C shows FXR mRNA expression (qPCR) in CCA tumors (n = 5) compared to normal human liver tissue (n = 20) and peripheral human liver tissue (n = 7) (San Sebastian cohort) (Man- Whitney test); 11D shows FXR mRNA expression (qPCR) in CCA tumors compared to matched-peripheral human liver tissue (n = 4) (san Sebastian cohort) (paired T-test).

As shown in FIGS. 12A-12D, TGR5 expression increases in CCA tumors, is higher around the pulmonary gate than in intrahepatic CCA, and is associated with perivascular invasion (FIG. 12A). 12B shows TGR5 mRNA microarray expression in total tissue of CCA tumors (n = 104) compared to surrounding human tissue (n = 60) (Copenhagen cohort) (Man-Whitney test). 12C shows CCA according to clinical-pathological parameters: anatomical location [peripheral occlusion (n = 36) or intrahepatic (n = 68)] and pericardial invasion [negative (n = 50) or positive (n = 42)]. TGR5 mRNA microarray expression in the whole tissue of the tumor is shown (Copenhagen cohort) (Man-Whitney test). 12C shows TGR5 mRNA expression (qPCR) in CCA tumors (n = 15) compared to surrounding human liver tissue (n = 27) (San Sebastian cohort) (Man-Whitney test). TGR5 mRNA expression (qPCR) (San Sebastian cohort) (Wilcoxon coincident-paired indicated assay test) in CCA tumors compared to matched-peripheral human liver tissue (n = 11) is shown in FIG. 12D.

As shown in FIGS. 13A and 13B, FXR expression is reduced and TGR5 expression is increased in CCA cell lines compared to normal human bile duct cells: FIG. 13A shows normal human bile duct cells (n = 6) and CCA cell lines (n, respectively). FXR mRNA expression (qPCR) at 5 and 6) (man-Whitney test or unpaired T-test), FIG. 13B shows normal human bile duct cells (n = 6) and CCA cell lines (n = 5 and TGR5 mRNA expression (qPCR) in 4) is shown (unpaired T-test or Man-Whitney test).

Chronic administration of the FXR agonist OCA in an ectopic CCA mouse model inhibited tumor growth compared to untreated animals, and this effect was associated with reduced PCNA and Ki67 expression in the tumors of treated animals. As shown in FIGS. 14A-14D, FXR agonist obeticholic acid (OCA) inhibited tumor growth in vivo, which was associated with decreased expression of proliferation, biliary tract and epithelial markers. EGI1 cells were injected subcutaneously into CD1 nude male mice. Once the tumor had grown, the tumor was implanted in the liver of CD1 nude male mice. Two weeks later, MRI analysis was performed and treatment administration started in the diet. Tumor growth was monitored at 1 and 2 months by MRI. 14A shows representative MRI and liver images of untreated control mice, OCA-treated mice and Compound (4) -treated mice. The bar-graph of FIG. 14B shows tumor volume fold-change quantified by MRI (control n = 8, OCA n = 6 and compound (4) n = 9 (Man-Whitney test, one side). MRNA expression levels of ectopic CCA intratumoral proliferation (ie Ki67 and PCNA), biliary tract (ie CK19) and epithelial (ie ZO-1) markers are shown: n = 5-8 (Man-Whitney or unpaired T) Representative immunohistochemical images of hepatic ectopic CCA tumor proliferative markers (ie Ki67 and PCNA) in mice treated or untreated with OCA or Compound (4) are shown in FIG. No effect on CCA tumor growth was observed in animals chronically treated with TGR5 agonist compound (4).

17 shows the effect of OCA and Compound (4) on hepatic xenograft tumor expression of proliferative markers. MRNA expression levels of proliferative (ie, Cdc25a, cyclin D1 and cyclin D3) markers in hepatic ectopic CCA tumors. n = 5-8 (man-whitney or unpaired T-test, one side).

In vitro, activation of FXR by OCA inhibited the proliferation and migration of CCA cells, and this event was associated with decreased mitochondrial energy metabolism (ie, reduced baseline breathing, maximal breathing, and ATP-associated breathing) compared to controls. It was related. In contrast, FXR activation did not affect the survival of CCA cells. As shown in FIGS. 15A-15D, FXR agonist OCA inhibits CCA cell proliferation in a dose-dependent manner and inhibits CCA cell migration associated with reduced mitochondrial metabolism in CCA cells, without inducing apoptosis. FIG. 15A shows CCA cells treated for 48 hours with 10 or 25 μM OCA (n = 5) compared to untreated control cells (n = 4), performed by flow cytometry using CFSE cell proliferating dye staining (FIG. Ie, proliferation of EGI1) (unpaired T-test). The analysis was performed with at least three independent experiments. MRNA expression of proliferation markers (ie Ki67, PCNA, cyclin D1 and cyclin D3) in CCA cells (ie EGI1) under OCA (25 μM) treatment for 3-6-12 hours compared to untreated control cells. n = 5-6, unpaired T-test on untreated control. 15B shows the results of migration analysis in CCA cells (ie EGI1). Representative microscopic images of wound-healing analysis at the indicated time points and conditions (ie, untreated control or OCA-treated) and corresponding quantification (n = 6 in each condition) (unpaired T-test). The analysis was performed in two independent experiments. Representative microscopic images of the transwell transfer chamber at 24 hours in untreated and OCA-treated cells. The analysis was performed once. 15C shows hippocampal oxygen consumption rate (OCR) using a mitochondrial stress test kit in untreated or untreated or treated with OCA (25 μM) with 3 hours pre-treatment (ie EGI1). Bar-graph of metabolic parameters calculated at OCR measurement. N = 11-12 (unpaired T-test) for each group. Analysis was performed in at least three independent experiments. FIG. 15D shows flow cytometry-based apoptosis analysis by staining Annexin V and propidium iodide in CCA cells treated with or without 10 or 25 μM OCA (ie, EGI1). Representative histogram and corresponding quantification of pooled data from two independent experiments, total n = 6-7 (unpaired T-test). The analysis was performed in three independent experiments.

In contrast, activation of TGR5 by compound (4) stimulated proliferation and migration of CCA cells, which event increased mitochondrial energy metabolism (ie increased baseline respiration, proton-leak and ATP-associated compared to control). Breathing). As shown in FIGS. 16A-16C, TGR5 agonist compound (4) slightly stimulates proliferation, migration and mitochondrial metabolism of CCA cells. 16A shows the proliferation of CCA cells (ie, EGI1) treated for 48 hours with 10 or 25 μM of compound (4) compared to untreated control cells, analyzed by flow cytometry using CFSE cell proliferation dye staining. Indicates. n = 5-6 (unpaired T-test). The analysis was performed with three independent experiments. Of proliferation markers (ie Ki67, PCNA, Cyclin D1 and Cyclin D3) in CCA cells (ie EGI1) under compound (4) (25 μM) treatment for 3-6-12 hours compared to untreated control cells. mRNA expression. n = 6, (unpaired T-test or Man-Whitney test on untreated control). 16B shows migration analysis in CCA cells (ie EGI1). Representative microscopic images and corresponding quantification of wound-healing assays at the indicated time points and conditions (ie, untreated control or compound (4) -treated). Pooled data from two independent experiments (n = 9 and 6 total) (unpaired T-test). Analysis was performed in at least three independent experiments. Representative microscopic images and corresponding quantification (n = 4 and 2) of the transwell transfer chamber at 24 hours in untreated and Compound (4) -treated cells. The analysis was performed in three independent experiments. 16C shows hippocampal oxygen consumption rate (OCR) using a mitochondrial stress test kit in CCA cells (ie, EGI1) untreated or treated with Compound (4) (25 μM) with 3 hours pre-treatment. Bar-graph of metabolic parameters calculated at OCR measurement. N = 11-12 (unpaired T-test) for each group. Analysis was performed in at least three independent experiments.

Claims (16)

Treating or preventing cancer in a subject in need thereof comprising administering a therapeutically effective amount of an FXR agonist selected from compound (1) or compound (2), or a pharmaceutically acceptable salt or amino acid conjugate thereof Way:

. The method of claim 1,
Said cancer is selected from hepatocellular carcinoma, cholangiocarcinoma, pancreatic cancer, kidney cancer, prostate cancer, esophageal cancer, breast cancer, gastric cancer, kidney cancer, salivary gland cancer, ovarian cancer, uterine cancer, bladder cancer, and lung cancer. The method according to claim 1 or 2,
The cancer is hepatocellular carcinoma. The method of claim 3,
The hepatocellular carcinoma from the group consisting of early stage hepatocellular carcinoma, non-metastatic hepatocellular carcinoma, primary hepatocellular carcinoma, advanced hepatocellular carcinoma, locally advanced hepatocellular carcinoma, metastatic hepatocellular carcinoma, remission hepatocellular carcinoma, or recurrent hepatocellular carcinoma The method chosen. The method according to any one of claims 1 to 4,
Wherein said FXR agonist is Compound (1) or a pharmaceutically acceptable salt thereof. The method of claim 5,
The FXR agonist is the sodium salt of compound (1). The method of claim 5,
Said FXR agonist is N, N- diethanamine salt of compound (1). The method according to any one of claims 1 to 4,
Wherein said FXR agonist is Compound (2) or a pharmaceutically acceptable salt or amino acid conjugate thereof. The method of claim 8,
Wherein said FXR agonist is a glycine conjugate of compound (2). The method of claim 8,
Said FXR agonist is a taurine conjugate of compound (2). The method of claim 8,
Wherein said FXR agonist is a sarcosine conjugate of compound (2). Comprising FXR agonists and pharmaceutically acceptable excipients selected from compound (1) or compound (2), or a pharmaceutically acceptable salt or amino acid conjugate thereof for treating or preventing cancer in a subject in need thereof Pharmaceutical Compositions:

. Kits for treating or preventing cancer in a subject in need thereof comprising compound (1) or compound (2), or a pharmaceutically acceptable salt or amino acid conjugate thereof:

. Use of an FXR agonist selected from compound (1) or compound (2), or a pharmaceutically acceptable salt or amino acid conjugate thereof in the manufacture of a medicament for the treatment or prophylaxis of cancer in a subject in need thereof:

. The method according to claim 1 or 2,
The cancer is cholangiocarcinoma. The method of claim 15,
Wherein said FXR agonist is Compound (1) or a pharmaceutically acceptable salt thereof.

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