CN110430882A - The method for the treatment of cancer - Google Patents

Abstract

The present invention relates to the methods for the cancer for treating or preventing subject in need, and this method includes giving the compound of the present invention of therapeutically effective amount.

Description

The method for the treatment of cancer

Background technique

Cancer is primarily characterized in the quantity increase for being originated from the abnormal cell of given normal tissue, abnormal cell to neighbouring The intrusion of tissue or malignant cell are diffused into regional nodes and distal part position (transfer) by lymph or blood.Clinical data and Molecular biology research show cancer be one change before slight tumour multi-step process, may be certain Under the conditions of progress to tumor and formed.

Primary carcinoma of liver is one of most common cancer forms in the world.There are two types of main Types for liver cancer: hepatocellular carcinoma (HCC) (also referred to as malignant hepatoma) and cholangiocellular carcinoma (also referred to as cholangiocarcinoma (CCA)).HCC is primary carcinoma of liver Most common form, and develop in liver cell.HCC is occurred mainly in male and patient with cirrhosis.HCC is generation The third-largest common cause of one of most common cancer and cancer related mortality within the scope of boundary.The disease is usually in clinical table The advanced stage of existing process is diagnosed.Therefore, only the patient of 10%-15% be curative surgical operation candidate.For Most of HCC patients, whole body chemotherapy or supporting treatment are main therapeutic choices.

On the contrary, the main hair in the small bile duct epithelial cell (i.e. bile duct cell) in liver of cholangiocellular carcinoma or bile duct cancer Exhibition.Such cancer is more common in women.The disease incidence of cholangiocarcinoma (CCA) worldwide constantly increases, and And have become the second largest most common primary carcinoma of liver.Since these tumours make a definite diagnosis late and its refractory property, CCA patient is shown Badness come-off.

Up to the present, can effectively treating cancer (such as HCC and/or CCA) medication amount it is limited.For example, part is controlled The patient with metastatic hepatocellular carcinoma or hepatocellular carcinoma for treating failure is usually only survived three to four months.Local treatment failure Metastatic hepatocellular carcinoma or hepatocellular carcinoma are substantially carried out constitutional treatment.Using Doxorubicin (tamoxifen of high dose with it is how soft Combined than star or EA-PFL (Etoposide, adriamycin, cis-platinum, fluorouracil and folinic acid)) it is an effective example.These The remission rate of drug can reach the level between 15% to 30%.However, since patients with hepatocellular carcinoma usually will appear cirrhosis Complication and other complication (such as oligoleukocythemia, decrease of platelet or hepatic disorder), therefore not can be carried out generalization Therapy.In addition, most of chemotherapeutants show limited effect and cannot significantly improve the survival of patient.Although Effort is carried out, but the bad clinical process of most of cancer patients is highlighted to more effective chemotherapeutic demand.

Cholangiocarcinoma is a kind of relatively rare tumour, and being classified as gland cancer, (a kind of is formationed body of gland secretes a large amount of mucoproteins Cancer).In the Western countries, the annual morbidity of cholangiocarcinoma be have 1-2 in every 100,000, but in the past few decades in, gallbladder Pipe cancer morbidity worldwide constantly rises.Unless both primary tumor and any metastatic tumor can pass through surgery hand Art is cut off completely, and otherwise cholangiocarcinoma is considered as that can not cure and lethal cancer rapidly.In addition to surgical operation, there is no potential Curative therapy, but most people has suffered from terminal illness in morbidity, and can not perform the operation in diagnosis.Usually By chemotherapy, radiotherapy and other palliative treatment controlling measurement cholangiocarcinoma patients, but can not cure.

Present invention accomplishes these needs.Therefore, the purpose of the present invention is to provide treat or prevent cancer (such as liver cell Cancer) and cholangiocarcinoma improved method.

Detailed description of the invention

Figure 1A is display compound 1-Na and control diet to 2 (Mdr2 of multi-drug resistance albumen^-/-) liver tumour in knock-out mice The bar chart of the influence of quantity.* the comparison of p < 0.01 control.

Figure 1B is display compound 1-Na and control diet to Mdr2^-/-Tumour reduces percentage (> 5mm diameter) in mouse Influence bar chart.

Fig. 2A is display OCA (compound 2) and control diet to Mdr2^-/-The influence of liver tumour quantity in knock-out mice Bar chart.

Fig. 2 B is display OCA and control diet to Mdr2^-/-Tumour reduces the influence of percentage (> 5mm diameter) in mouse Bar chart.

Fig. 3 A is display compound 1-Na and control diet to farnesoid X receptor (FXR^-/-) liver tumour quantity in mouse The bar chart of influence.

Fig. 3 B is display compound 1-Na and control diet to FXR^-/-Tumour reduces percentage (> 5mm diameter) in mouse The bar chart of influence.

Fig. 4 is display compound 1-Na and control diet to FXR^-/-Liver tumour quantity reduces the influence of percentage in mouse Bar chart.

Fig. 5 A is display compound 1-Na and control diet to Mdr2^-/-Liver/weight ratio reduces the shadow of percentage in mouse Loud bar chart.* the comparison of p < 0.01 control.

Fig. 5 B is display compound 1-Na and control diet to FXR^-/-Liver/weight ratio reduces the shadow of percentage in mouse Loud bar chart.

Fig. 6 A is display compound 1-Na and control diet to Mdr2^-/-And FXR^-/-Alanine aminotransferase (ALT) in mouse The bar chart of horizontal influence.* the comparison of p < 0.01 control.

Fig. 6 B is display compound 1-Na and control diet to Mdr2^-/-And FXR^-/-Aspartate transaminase in mouse (AST) bar chart of horizontal influence.* the comparison of p < 0.01 control.

Fig. 7 A is display compound 1-Na and control diet to Mdr2^-/-And FXR^-/-Fibroblast growth factor in mouse The bar chart of the influence of the ileum gene expression of 15 (Fgf15).* the comparison of p < 0.01 control.

Fig. 7 B is display compound 1-Na and control diet to Mdr2^-/-And FXR^-/-Small heterodimeric partner in mouse (Shp) bar chart of the influence of ileum gene expression.* the comparison of p < 0.01 control.

Fig. 8 is display compound 1-Na and control diet to Mdr2^-/-And FXR^-/-Cholesterol 7α-hydroxylase in mouse (cyp7a1) bar chart for the influence lowered.* the comparison of p < 0.01 control.

Fig. 9 A is display compound 1-Na and control diet to Mdr2^-/-And FXR^-/-Small heterodimeric partner in mouse (Shp) bar chart of the influence of liver gene expression.* the comparison of p < 0.01 control.

Fig. 9 B is display compound 1-Na and control diet to Mdr2^-/-And FXR^-/-Bile salt rear pump (Bsep) in mouse Liver gene expression influence bar chart.* the comparison of p < 0.01 control.

Figure 10 A is display compound 1-Na and control diet to Mdr2^-/-The bar shaped of the influence of total serum bile acid in mouse Figure.* the comparison of p < 0.01 control.

Figure 10 B is display compound 1-Na3 and control diet to FXR^-/-The bar shaped of the influence of total serum bile acid in mouse Figure.

Figure 11 A shows that compared with people around's tissue, the FXR mRNA microarray in the entire tissue of CCA tumour is expressed.

Figure 11 B shows the FXRmRNA microarray table in the entire tissue according to the CCA tumour of Tumor Differentiation hierarchal grouping It reaches.

Figure 11 C shows that the FXRmRNA compared with normal human liver tissue and surrounding human liver tissue, in CCA tumour is expressed (qPCR)。

Figure 11 D shows that the FXR mRNA compared with matched surrounding human liver tissue, in CCA tumour expresses (qPCR).

Figure 12 A shows that compared with people around's tissue, the TGR mRNA microarray in the entire tissue of CCA tumour is expressed.

Figure 12 B is shown under Clinicopathological Parameters (anatomical position and neural surrounding wetting), the entire tissue of CCA tumour In TGR5 mRNA microarray expression.

Figure 12 C shows that compared with human liver tissue around, the TGR5 mRNA in CCA tumour expresses (qPCR).

Figure 12 D shows that the TGR5 mRNA compared with matched surrounding human liver tissue, in CCA tumour expresses (qPCR).

Figure 13 A shows that the FXR mRNA in normal person's bile duct cell and CCA cell line expresses (qPCR).

Figure 13 B shows that the TGR5 mRNA in normal person's bile duct cell and CCA cell line expresses (qPCR).

Figure 14 A shows the representative MRI for the mouse that untreated control mice, the mouse of OCA processing and compound 4 are handled And liver image.

Figure 14 B is the bar chart shown through MRI quantitative gross tumor volume multiple variation.

Figure 14 C shows proliferation mark (i.e. Ki67 and PCNA), biliary tract mark (i.e. CK19) and upper in hepatocyte in situ CCA tumour The mRNA expression of leather mark will (i.e. ZO-1).

Figure 14 D shows in the liver original position CCA tumour of untreated mouse or the mouse handled with OCA or compound 4 and is proliferated Indicate the representative immunohistochemistry image of (i.e. Ki67 and PCNA).

Figure 15 A is shown compared with untreated control cell, and the mRNA expression of mark is proliferated after OCA processing, and (CFSE cell Proliferation is used by flow cytometry with the proliferation of the OCA CCA cell (i.e. EGI1) for handling 48h with 10 or 25 μM Dyeing carries out).

Figure 15 B shows that the migration in CCA cell (i.e. EGI1) measures and wound healing measures and accordingly quantitative representativeness Micro-image and the representative micro-image of 24 hours transwell migration room.

Figure 15 C be shown in CCA cell (i.e. EGI1) using mitochondria stress test kit hippocampus (Seahorse) The bar chart of consumption rate (OCR) and the metabolizing parameters calculated in OCR measurement.

Figure 15 D is shown in the CCA cell (i.e. EGI1) of untreated or OCA processing with 10 or 25 μM, joins egg by film The representative histogram of the Apoptosis measurement and merging data based on flow cytometry of white V and propidium iodide and It is corresponding quantitative.

Figure 16 A shows that compared with untreated control cell, the mRNA that mark is proliferated after the processing of compound 4 expresses water It is flat, and with compound 3 with the proliferation of the CCA cell (i.e. EGI1) of 10 or 25 μM of processing 48h.

Figure 16 B shows that migration measurement, wound healing measurement and accordingly quantitative representativeness in CCA cell (i.e. EGI1) are shown The migration room transwell for 24 hours and accordingly quantitative representative in the cell that micro- image and untreated and compound 4 are handled Property micro-image.

Figure 16 C is shown in CCA cell (i.e. EGI1) that is untreated or being handled with (25 μM) of compound 4 and uses mitochondria It stress the hippocampus consumption rate (OCR) of test kit and the bar chart of the metabolizing parameters calculated in OCR measurement.

Figure 17 shows proliferation mark (i.e. Cdc25a, cyclin D1 and the cell cycle in hepatocyte in situ CCA tumour Protein D 3) mRNA expression.

Summary of the invention

This application involves the method for the cancer for treating or preventing subject in need, this method includes giving to treat effectively Farnesoid X receptor (FXR) agonist of amount.In one embodiment, which is compound 1 or compound 2:

Or its pharmaceutically acceptable salt or amino acid conjugate.

In one embodiment, which is selected from the group, which is made up of: hepatocellular carcinoma, cancer of pancreas, kidney, preceding Column gland cancer, the cancer of the esophagus, breast cancer, gastric cancer, kidney disease, salivary-gland carcinoma, oophoroma, carcinoma of uterine body, bladder cancer and lung cancer.At one In embodiment, which is hepatocellular carcinoma.In one embodiment, which is cancer of pancreas.In one embodiment, the cancer It is kidney.In one embodiment, which is prostate cancer.In one embodiment, which is the cancer of the esophagus.In a reality It applies in example, which is breast cancer.In one embodiment, which is gastric cancer.In one embodiment, which is kidney Disease.In one embodiment, which is salivary-gland carcinoma.In one embodiment, which is oophoroma.In one embodiment In, which is carcinoma of uterine body.In one embodiment, which is lung cancer.

In one embodiment, which is compound 1 or its pharmaceutically acceptable salt.In another implementation In example, which is the sodium salt (i.e. compound 1-Na) of compound 1.In another embodiment, the FXR agonist It is the N of compound 1, N- diethyl alkanamine (i.e. compound 1-DEA).

In another embodiment, which is compound 2 or its pharmaceutically acceptable salt or amino acid conjugation Object.In one embodiment, which is the glycine conjugates of compound 2.In one embodiment, FXR excitement Agent is the taurine conjugate of compound 2.In one embodiment, which is the sarcosine conjugate of compound 2.

The invention further relates to compound 1 or its pharmaceutically acceptable salt or compound 2 or its can pharmaceutically connect Purposes of the salt or amino acid conjugate received in the drug for preparing the cancer for treating or preventing subject in need.

The invention further relates to compound 1 or its pharmaceutically acceptable salt and compound 2 or its is pharmaceutically acceptable Salt or amino acid conjugate, for being used in the cancer for treating or preventing subject in need.

The invention further relates to pharmaceutical composition, the pharmaceutical composition include compound 1 or its pharmaceutically acceptable salt and Compound 2 or its pharmaceutically acceptable salt or amino acid conjugate (its cancer for being used to treat or prevent subject in need Disease) and pharmaceutically acceptable excipient.

The invention further relates to treat or prevent the kit of the cancer of subject in need, the kit packet Containing compound 1 or its pharmaceutically acceptable salt and compound 2 or its pharmaceutically acceptable salt or amino acid conjugate.

Specific embodiment

The present invention is at least partially based on following discovery: compound 1 and 2 can be treated effectively in the animal model of prediction cancer Cancer.As described in following instance, ladies and gentlemen inventor has found the mouse mould that the compound of the present invention inhibits spontaneous hepatocellular carcinoma to occur Tumour growth in type.

Definition

For convenience's sake, certain terms used in this specification, example and the appended claims have been concentrated herein.

As used herein term " cancer " refers to so that there are any diseases characterized by cancerous tissue in subject.

As it is used herein, " cancerous tissue " refers to the tissue comprising malignant cell, show excrescent Cell and/or excessive proliferated cell.Cancerous tissue can be from tissue or rise source organ primary malignant tumor or it can To be metastatic malignant tumour, being grown in is not in the bodily tissue in primary tumor source.

As it is used herein, term " tumour " may include the cancer of solid tumor or haematological origin.In some embodiments, Tumour is characterized in that it invades surrounding tissue, is transferred to body other parts and/or the ability by its angiogenic activity. Exemplary oncologic is by hepatocellular carcinoma, gastric cancer, kidney disease, prostate cancer, adrenal, cancer of pancreas, breast cancer, bladder cancer, saliva Gland cancer, oophoroma, carcinoma of uterine body and lung cancer generate.

As it is used herein, term " invasive " refers to that cell, one group of cell or malignant tumour are spread from a position To the process of adjacent parts.

As used herein term " metastatic " refers to that cell, one group of cell or malignant tumour are diffused into from a position The not process at the position adjacent with first position.

As it is used herein, " hepatocellular carcinoma ", " HCC " and " malignant hepatoma " is used interchangeably, and refer to source From the primary and secondary (metastatic) tumour of hepatic tissue.As used herein term " intractable hepatocellular carcinoma " refer to as Advantageous reaction is made in lower hepatocellular carcinoma, the non-confrontational oncotherapy of the hepatocellular carcinoma.It is therefore, as used herein that " treatment is difficult Hepatocellular carcinoma " refer to following hepatocellular carcinoma, which does not make advantageous reaction to treatment or generates resistance to treatment, or Person sends out or recurs again after alternatively generating advantageous reaction to treatment.

As it is used herein, " cholangiocarcinoma " or " CCA " or " cholangiocellular carcinoma " or bile duct cancer are by originating from gallbladder Juice is discharged into the cancer shape of epithelial cell (or cell of display epithelial differentiation feature) composition of the mutation of the bile duct of small intestine from liver Formula.A part of CCA occurs under the liver condition of cholestasis.Although accumulation cannot directly induce carcinogenic work in the liver of bile acid With, but they can be by promoting bile duct cell proliferation carcinogenic to promote with inflammation and by reduction FXR dependence chemoproection Effect.

As it is used herein, term " compound 1 " refers to

It is also referred to as -3 α of 6 α-ethyl, and -5 β-cholane -23- hydrogen sulfate drops in 7 α, 23- trihydroxy -24-." compound 1- Na " or " 1-Na " (it is also referred to as -3 α of 6 α-ethyl, and -5 β-cholane -23- sodium sulphate drops in 7 α, 23- trihydroxy -24- ") it is interchangeable It uses, and refers to the sodium salt of compound 1.As it is used herein, (it is also referred to as 6 by " compound 1-DEA " or " 1-DEA " - 5 β-cholane -23- sulfate N, N- diethyl ethane amine drops in α-ethyl -3 α, 7 α, 23- trihydroxy -24- ") it is used interchangeably, and And refer to the N of compound 1, N- diethyl ethane amine salt.The structure of compound 1-Na and compound 1-DEA are as follows.

" compound 2 " refers to as used herein

It is also referred to as Austria's shellfish cholic acid (OCA), 6-ECDCA, 6- α-ethyl chenodeoxycholic acid or 6 α-ethyl -3 α, 7 α-two - 5 β of hydroxyl-cholane -24- acid.

As it is used herein, " compound 3 " refers to

It is also referred to as 3 α, 7 α, -6 α of 11 β-trihydroxy--5 β of ethyl-cholane -24- acid.

As it is used herein, " compound 4 " refers to

It is also referred to as 6 α-ethyl -23 (S)-methyl -3 α, 7 α, -5 β of 12 α-trihydroxy-cholane -24- acid.United States Patent (USP) Compound 4 described in numbers 8,114,862 is TGR5 regulator.In one embodiment, TGR5 regulator is agonist.

Term " TGR5 regulator " means any compound with TGR5 acceptor interaction.Interaction is not limited to serve as The antagonist of TGR5 receptor, agonist, partial agonist or inverse agonist compound.

In general, term " agonist " means to enhance another molecule or the active compound of acceptor site.It is defined according to classics, Agonist (either positive structure site agonist (orthosteric agonist), allosteric agonist, inverse agonist or collaboration Agonist (co-agonist)) there is characteristic in conjunction with receptor, change its receptor status and lead to biological effect.Therefore, swash The dynamic characteristic for being defined as agonist or ligand generation biological effect.More particularly, TGR5 agonist is the receptor in conjunction with TGR5 Ligand or compound, and make in the cell of expressed receptor the concentration increase at least 20% of 3'5'-AMP (cAMP).

As used herein " the compound of the present invention " cover compound 1,1-Na, 1-DEA, 2 and 3 or its pharmaceutically Acceptable salt or amino acid conjugate.

As used herein term " treatment " refers to successful treatment or improves any mark of cancer.Treatment may include Such as mitigate or alleviate cancer one or more symptoms severity or its may include reduce patient experience disease The frequency of the symptom of disease, defect, obstacle or unfavorable illness etc.." treatment " can also refer to a part (example for reducing or eliminating body Such as cell, tissue or body fluid, such as blood) illness.

As it is used herein, term " prevention " refers in partially or completely prevention individual or group or a part of body Cancer in (such as cell, tissue or body fluid, such as blood).Term " prevention " does not require in the group of entire treatment individual or a Prevent disease or illness completely in the cell of body, tissue or liquid.As used herein term " treatment or prevention " makes to instruct It causes treatment to a certain degree or improves the method for cancer, and consider the series of results for being directed to the purpose, including but not limited to Complete pre- anti-cancer.

As used herein phrase " therapeutically effective amount " refers to comprising being enough to make tumor regression and/or reducing the tumour Growth rate (as compacting tumour growth) or prevent or delay the undesirable cell Proliferation of other in cancer amount effective quantity.One In a little embodiments, effective quantity is the amount for being enough to postpone cancer development.In some embodiments, effective quantity is to be enough to prevent or delay Recurrent amount.Effective quantity can be given with one or many give.In the case where HCC or CRC, drug or composition Effective quantity may is that (i) reduce tumour cell quantity；(ii) reduce tumor size；(iii) inhibit, postpone, in certain journey Slow down on degree and preferably stops cancer cell infiltration into peripheral organ；(iv) inhibit (that is, slow down to a certain extent and Preferably stop) metastases；(v) inhibit tumour growth；(vi) it prevents or delays the generation of tumour and/or sends out again, and/or (vii) alleviate one or more symptoms relevant to cancer to a certain extent.

As used herein term " scheme " refers to for being administered and/or periodically giving the compound of the present invention for controlling Treat the scheme of cancer.Scheme may include actively giving phase and rest period as known in the art.Actively giving the phase is included in The compound of the present invention is given in the time course of restriction comprising such as number of composition administration and time.In some sides In case, it may include one or more rest periods, do not give compound actively in the rest period, and in some cases, it should Rest period includes that the effect of such compound may be the smallest period.

As it is used herein, " combination treatment " refer to the compound of the present invention can with another therapeutic agent to It gives." with ... combine " refer to other than giving a kind for the treatment of mode, another treatment mode is also given, such as same in addition to giving Outside the compound of one subject's invention as described herein, another therapeutic agent is also given.Therefore, " with ... combine " refer to A kind for the treatment of mode is given before, during or after delivering the second treatment mode to subject.

As it is used herein, it is not biologically or other undesirable materials of aspect that " pharmaceutically acceptable ", which refers to, Material, for example, the material can be mixed into the pharmaceutical composition that patient gives, without causing any undesirable biological effect Or it interacts in harmful manner with any other component of the composition comprising it.Pharmaceutically acceptable carrier or figuration Agent meets the required standard of toxicology and manufacture test and/or is included in by food and drug administration (U.S.Food And Drug administration) preparation Inactive Ingredient Guide (Inactive Ingredient Guide) in.

When referring to measurable value (such as amount, duration etc.), as used herein term " about " is intended to refer to ± 20% or ± 10% variation (in some embodiments ± 5%, in some embodiments ± 1%, and some of definite value ± 0.1% in embodiment) because such variation is suitable for practicing disclosed method, or suitable for manufacturing and using disclosed change Close object and in claimed method.

The method for the treatment of cancer

The present invention is at least partially based on following discovery: the compound of the present invention is effectively controlled in the mouse model of prediction human cancer Treat cancer.Therefore, this application involves the method for the cancer for treating or preventing subject in need, this method includes giving to treat A effective amount of FXR agonist, the FXR agonist are selected from the group, which is made up of: compound 1,1-Na, 1-DEA, 2 and 3:

Or its pharmaceutically acceptable salt or amino acid conjugate.In one embodiment, which is compound 1.In one embodiment, which is compound 1-Na.In another embodiment, which is compound 1-DEA.In one embodiment, which is compound 2.In one embodiment, which is compound 3。

In method described herein, exemplary cancers are selected from the group, which is made up of: hepatocellular carcinoma, cholangiocarcinoma, Cancer of pancreas, prostate cancer, the cancer of the esophagus, breast cancer, gastric cancer, kidney disease, salivary-gland carcinoma, oophoroma, carcinoma of uterine body, bladder cancer and lung Cancer.Appropriate therapeutic scheme for cancer depend on the cell in tumour institute source type, it is pernicious by stages with severity, with And facilitate the hereditary disorder of tumour.

The degree of cancer staging System describe cancer progression.In general, these Staging Systems describe the journey of cancer diffusion Degree, and the patient with similar prognosis and treatment is divided into same group by stages.Generally, for become it is invasive or turn The tumour of shifting, prognosis are poor.In a type of Staging System, case is divided into four-stage, with Roman number I to IV table Show.In the I phase, cancer is usually part, and usually recoverable.II phase and IIIA phase cancer usually more advanced stage, and can Surrounding tissue can have been invaded and diffused to lymph node.IV phase cancer includes having diffused to the metastatic carcinoma at outside lymph node position Disease.

Another Staging System is TNM stage, it represents classification: tumour, section and transfer.In this system, according to each The severity of a classification describes malignant tumour.For example, T classifies the degree from 0 to 4 of primary tumo(u)r, wherein 0 representative does not have Invasive active malignant tumour, and 4 represent the malignant tumour for extending from initial site and invading other organs.N will drench The degree classification of involvement is fawned on, wherein 0 represents the malignant tumour of not lymph node involvement, and 4 represent with extensive lymph node The malignant tumour of involvement.M classifies the degree from 0 to 1 of transfer, wherein 0 represents the malignant tumour without transfer, and 1 represents tool There is the malignant tumour of transfer.

The variation of these Staging Systems or these Staging Systems or other suitable Staging Systems can be used for describing tumour. According to cancer by stages and feature, the selection for being available for treating cancer are seldom.Treatment is controlled including surgical operation, with Sorafenib Treatment and targeted therapies.In general, surgical operation is the First Line treatment for early stage localized cancer.Other systemic therapy can For treating invasive and metastatic tumo(u)r.

According to an aspect of the invention, there is provided the method for treating hepatocellular carcinoma (or malignant hepatoma).It is special Not, this method includes that the subject in need of hepatocellular carcinoma is suffered from the compound of the present invention treatment of therapeutically effective amount. That is, the present invention relates to the compound of the present invention the hepatocellular carcinoma with hepatocellular carcinoma is accredited as or is diagnosed as treating in preparation Purposes in the drug of patient.In individual embodiment, treatment method optionally includes diagnosis or identification patient is thin with liver The step of born of the same parents' cancer.Then identified patient is treated or given with the compound of the present invention of therapeutically effective amount.Liver cell Cancer can be diagnosed with any routine diagnostic method known in the art, including the test of ultrasound, CT scan, MRI, alpha-fetoprotein, The screening of des- γ carboxyl factor and biopsy.

The present invention also provides the method for treating intractable hepatocellular carcinoma, this method includes the present invention with therapeutically effective amount Compound treatment be accredited as the patient with intractable hepatocellular carcinoma.In certain embodiments, patient is tired with treatment Difficult hepatocellular carcinoma, the treatment include one or more drugs selected from the group below, which is made up of: Sorafenib, Rui Ge Non- Buddhist nun, anthracycline (such as Doxorubicin, daunorubicin, epirubicin, idarubicin), platinum agent (such as cis-platinum, carboplatin, Austria Husky benefit platinum, picoplatin), 5-FU and capecitabine.The invention further relates to the compound of the present invention in preparation for treating intractable liver Purposes in the drug of cell cancer, such as one or more drugs selected from the group below be intractable hepatocellular carcinoma, The group is made up of: Sorafenib, Rui Gefeini, anthracycline (such as Doxorubicin, daunorubicin, epirubicin, Yi Da Than star), platinum agent (cis-platinum, carboplatin, oxaliplatin, picoplatin), 5-FU and capecitabine.

In order to detect intractable hepatocellular carcinoma, the resistance for receiving the patient of initial treatment, unresponsiveness can be carefully monitored Or the sign of recidivity hepatocellular carcinoma.This can realize that this is initial by monitoring the cancer of patient to the response of initial treatment Treatment for example may include one or more drugs selected from the group below, which is made up of: Sorafenib, Rui Gefeini, more It is soft than star, daunorubicin, epirubicin, idarubicin, cis-platinum, carboplatin, oxaliplatin, picoplatin, 5-FU, Tegafur and Ka Peita Shore.Any suitable method that cancer can practice the response, shortage response or recurrence of initial treatment by this field determines. For example, this can be realized by assessment tumor size and quantity.The increase of tumor size or alternatively tumour quantity Increase shows that recurrence is not reacted chemotherapy or had occurred and that tumour.Can according to Therasse et al., " RECIST " mark being described in detail in J.Natl.Cancer Inst. [National Cancer Institute magazine] 92:205-216 (2000) Standard is measured.

Yet other aspects according to the present invention are provided for preventing or delaying hepatocellular carcinoma (or hepatocellular carcinoma) hair Make or prevent or delay the recurrent method of hepatocellular carcinoma, this method includes being controlled with the compound of the present invention of prevention effective dose Treat the patient for needing to prevent or delay.

The known risk for suffering from hepatocellular carcinoma with hepatitis B or hepatitis C infection or the subject with cirrhosis increases Add.In addition, suffering from acute and chronic Waldenstrom I type syndrome (acute intermittent porphyria, porphyria cutanea tarda, heredity excrement porphin Quinoline disease, ambiguity porphyria) and people's risk for suffering from hepatocellular carcinoma of I type tyrosinemia also increase.These people can be made The candidate of the method for the invention of hepatocellular carcinoma breaking-out is prevented or delayed with the compound of the present invention of prevention effective dose.This Outside, the patient with hepatocellular carcinoma family history can also be identified, to prevent or delay what hepatocellular carcinoma was broken out using of the invention Method.In order to prevent or delay sending out again for hepatocellular carcinoma, having treated and be in paracmasis or the stationary phase of being in or get nowhere The patients with hepatocellular carcinoma of state can be treated with the compound of the present invention of prevention effective dose, thin effectively to prevent or delay liver Born of the same parents' cancer is sent out again or is recurred.

According to the one aspect of present disclosure, the method for treating cholangiocarcinoma (CCA) is provided.Particularly, this method packet Include the subject in need that cholangiocarcinoma is suffered from the compound of the present invention treatment of therapeutically effective amount.That is, the present invention relates to this The compound of invention is in preparation for treating the purposes in the drug for being accredited as or being diagnosed as the cholangiocarcinoma patients with cholangiocarcinoma. In individual embodiment, treatment method optionally includes the step of diagnosis or identification patient are with cholangiocarcinoma.Then with treatment Identified patient is given in a effective amount of the compound of the present invention treatment.Cholangiocarcinoma can be with known in the art any Routine diagnostic method diagnosis, including ultrasound, CT scan, MRI, carcinomebryonic antigen (CEA) and Carbohydrate Antigens 19-9 (CA19- 9) screening and biopsy.

Yet other aspects according to the present invention are provided for preventing or delaying cholangiocarcinoma breaking-out or preventing or delaying The recurrent method of cholangiocarcinoma, this method include needing to prevent or delay with the compound of the present invention treatment of prevention effective dose Patient.

In order to prevent or delay sending out again for cholangiocarcinoma, having treated and be in paracmasis or the stationary phase of being in or get nowhere The cholangiocarcinoma patients of state can be treated with the compound of the present invention of prevention effective dose, effectively to prevent or delay cholangiocarcinoma Send out or recur again.

According to another aspect of the present invention, the method for inhibiting CCA cell Proliferation and migration with FXR agonist is provided.

According to another aspect of the present invention, it provides and inhibits CCA cell with the compound of the present invention of therapeutically effective amount The method of proliferation and migration.According to another aspect of the present invention, the method for inhibiting CCA progress is provided, this method includes using The compound of the present invention of therapeutically effective amount treats patient in need.

One embodiment is the method for treating cancer of pancreas by giving the compound of the present invention of therapeutically effective amount.Another Embodiment is the method for treating prostate cancer by giving the compound of the present invention of therapeutically effective amount.Another embodiment is logical Cross the method for giving the compound of the present invention treatment kidney of therapeutically effective amount.Another embodiment is treated effectively by giving The method of the compound of the present invention treatment prostate cancer of amount.It is by giving therapeutically effective amount in still another embodiment The compound of the present invention treatment the cancer of the esophagus method.It is the sheet by giving therapeutically effective amount in still another embodiment The method of the compound treatment breast cancer of invention.One embodiment is controlled by giving the compound of the present invention of therapeutically effective amount The method for treating gastric cancer.It in another embodiment, is to treat kidney disease by giving the compound of the present invention of therapeutically effective amount Method.It is to treat salivary-gland carcinoma by giving the compound of the present invention of therapeutically effective amount in still another embodiment Method.It is the method for treating oophoroma by giving the compound of the present invention of therapeutically effective amount in still another embodiment. One embodiment is the method for treating carcinoma of uterine body by giving the compound of the present invention of therapeutically effective amount.In another implementation It is the method for treating bladder cancer by giving the compound of the present invention of therapeutically effective amount in example.In still another embodiment, It is the method for treating lung cancer by giving the compound of the present invention of therapeutically effective amount.

In the case where the compound of the present invention can be used for treating cancer as described herein, such compound can be with the second medicine Agent is given jointly.The second medicament that can be used for treating the method for cancer provided herein may include the anticancer of any known class Agent, as such as radiotherapy, operation, alkylating agent, antimetabolite, anthracycline, camptothecine, vinca alkaloids, taxane Class or platinum and other antitumor agents as known in the art.This anticancer agent and antitumor agent classification be it is known in the art, And it is used according to its general and common meaning.

Exemplary anticancer agent includes but is not limited to: ABRAXANE；Abiraterone；ace-11；Aclarubicin；Acivicin； Hydrochloric acid acodzole；Acronine；D actinomycin D；Acyl group husband's text；Gland cyclopentanol；Adozelesin；Adriamycin；Aldesleukin； All-trans retinoic acid (ATRA)；Hemel；Ambamustine；Ambomycin；Acetic acid Ametantrone；Amidol gram this (amidox)；Amifostine；Aminoglutethimide；Amino-laevulic acid；Amrubicin；Amsacrine；Anagrelide；Anastrozole；Punching Lotus lactone；Antarelix；Anthramycin；Aphidicolin glycine；Apurinic nucleic acid；ara-CDP-DL-PTBA；Arginine Deaminase；ARRY-162；ARRY-300；ARRY-142266；AS703026；Asparaginase；Asperline；A Sulai crin (asulacrine)；Atamestane；Atrimustine；Avastin；A Naisiting (axinastatin) 1；A Naisiting (axinastatin)2；A Naisiting (axinastatin) 3；Azasetron；Azalomvcin；Azatyrosine；Azacitidine； AZD8330；Azetepa；Azotomycin；Ba Lanuo (balanol)；Batimastat；BAY11-7082；BAY 43-9006；BAY 869766；Bendamustine；Benzo chlorin；Benzodepa；Benzoyl Staurosporine (benzoylstaurosporine)；β-alethine；Sub- Aclacinomycin B；Betulinic acid；B-FGF inhibitor；Bicalutamide； Bisantrene；Double aziridinyl spermine；Bisnafide；Two methanesulfonic acid bisnafides；Bistratene A；Bisantrene hydrochloride；It is rich next Mycin；Bleomycin Sulphate；Busulfan；Bizelesin；Bu Ruifulaite (breflate)；Bortezomib；Brequinar sodium；Bromine Found bright (bropirimine)；Budotitane；Cystathionine；Bryostatin；Act-C；Calusterone；Calcipotriol； Ka Futading C；Camptothecin derivative；Capecitabine；Formamide-amino-triazole；Carboxyamido triazole；CaRest M3； CARN 700；Caracemide；Carbetimer；Carboplatin；Carmustine；Carubicin hydrochloride (carubicin hydrochloride)； Carzelesin；Grain tree spermine；Cecropin B；Cedefingol；Celecoxib；Cetrorelix；Chlorin；Chloro-quinoxaline sulfonamide； Cicaprost；Chlorambucil；Chlorine melts albumen；Cirolemycin；Cis-platinum；CI-1040；Along porphyrin；Cladribine；Clomifene class Like object；Clotrimazole；Collins's moldin A；Collins's moldin B；Combretastatin A-4 4；Combretastatin analog；That Green of health (conagenin)；Kang Baixiting (crambescidin) 816；Crisnatol；The horizontal sour crisnatol of first (crisnatolmesylate)；Cryptophycin 8；Cryptophycin A derivative；Draw element A in library；Penta anthraquinone of ring；Match gram Alexandre Desplat (cycloplatam)；It is new (cypemycin) to match Pumi；Cyclophosphamide；Cytarabine；Cytarabine octadecyl phosphate；It is molten Cell factor；Hexestryl diphosphate；Dacarbazine；Dactinomycin D；Daunorubicin；Daunorubicin hydrochloride；Decarbazine；Up to former times list It is anti-；Dasatinib；Decitabine；Dehydrogenated membrane ectexin B；Deslorelin；Dexamethasone；Right ifosfamide；Dexrazoxane；It is right Verapamil；Dexormaplatin；Dezaguanine；Methanesulfonic acid Dezaguanine (dezaguaninemesylate)；Diaziquone；Film ascidian Plain B；Di Duokesi (didox)；Diethyl removes first spermine；5 azacytidine of dihydro；Dihydro taxol；9- dioxa moldin；Connection Benzene spiromustine；Docosanol；Dolasetron；Docetaxel；Doxorubicin；Doxorubicin hydrochloride；Deoxyfluorouridine；Bend Lip river former times It is fragrant；Droloxifene citrate；Dromostanolone propionate；Dronabinol；Duazomycin；More new SA of card rice；Ebselen；Yi Kaomosi Spit of fland；Edelfosine；Edrecolomab；Edatrexate；Hydrochloric acid Eflornithine (eflornithine hydrochloride)；According to Lip river Ni Sai；Elemene；Emitefur；Elsamitrucin；Enloplatin；Enpromate；Epipropidine；Epirubicin；Epirubicin hydrochloride； Epristeride；Erbulozole；Eribulin；Hydrochloric acid Aesop compares star；Estramustine；Estramustine phosphate sodium；Etanidazole；It relies on Moor glycosides；Etoposide phosphate；Etoprine；Exemestane；Fadrozole；Carbazole hydrochloride；Fazarabine；Suwei A amine；Non- lattice Take charge of pavilion；Finasteride；Flavopiridol (flavopiridol)；Flezelastine；Not Lars spy human relations (fluasterone)；Floxuridine (floxuridine)；Fludarabine phosphate；Fludarabine；Fluorine daunorubicin hydrochloride；Forfenimex；Formestane；Fluorine urine is phonetic Pyridine；Floxuridine (floxouridine)；Flurocitabine；Fosquidone；Fostriecin sodium；Fostriecin；Fotemustine；Gadolinium is for sand Woods；Gallium nitrate；Galocitabine；Ganirelix；Gelatinase inhibitor；Gemcitabine；Geldanamycin；gossyphol；GDC- 0973；GSK1120212/ Trimetinib；Trastuzumab；Hydroxycarbamide；hepsulfam；Heregulin；Hexamethylene bisacetamide；Hypericum Chinense Element；Ibandronic acid；Buddhist nun is replaced according to Shandong；Idarubicin；Idarubicin hydrochloride；Ifosfamide；Bank phosphamide；Ilmofosine；Isopropyl Platinum；Idoxifene；Idramantone；Ilmofosine；Ilomastat；Imidazo acridone；Imatinib (for example, GLEEVEC)； Imiquimod；iniparib(BSI201)；Iobenguane；Iododoxorubicin；Ipomeanol；Irinotecan；Irinotecan hydrochloride, Aesop It is fixed to draw；Different gal azoles (isobengazole)；Different high halichondrins (isohomohalicondrin) B；Itasetron；Yi Mofu It is pungent；Interleukins IL-2 (including recombinant interleukin II；Or rlL.sub.2)；Intederon Alpha-2a；Interferon Alpha-2b；It is dry Disturb plain α-n1；Alferon N；Interferon beta-1a；Gamma interferon 1-b；jasplakinolide；Sea slug extract (kahalalide)F；Triacetic acid stratiform element N；Lanreotide；Come Na meter Xin (leinamycin)；Lenograstim；Sulfuric acid mushroom is more Sugar；Lay is general appropriate this new (leptolstatin)；Letrozole；Enantone (leuprorelin)；Levamisol；Lenalidomide；Pleasure is cut down For Buddhist nun；Liarozole；lissoclinamide 7；Lobaplatin；Lombricine；Lometrexol；Lonidamine；Losoxantrone；Cut down him in Lip river Spit of fland；Loxoribine；Lurtotecan；For sarin lutetium；Lisofylline；Lanreotide acetate；Lapatinib；Letrozole；Folinic acid；Bright third Rayleigh acetate；Liarozole hydrochloride；Lometrexol sodium；Lomustine；Losoxantrone hydrochloride；Pomalidomide (pomalidomide)；LY294002；Maitansine；Make sweet carbohydrase element A；Marimastat；Masoprocol；Mammary gland silk presses down albumen； Matrilysin inhibitor；Menogaril；Mei Balong；Meterelin；Methioninase；Metoclopramide；MIF inhibitor； Mifepristone；Miltefosine；Mirimostim；Mitoguazone；Mitolactol；Mitonafide；Mitoxantrone；Mofarotene；Not Plast pavilion；Mopidamol；mycaperoxide B；myriaporone；Maitansine；Mustine hydrochlcride；Megestrol acetate；Acetic acid Melengestrol；Melphalan；Purinethol；Methotrexate (MTX)；Methotrexate sodium；Metoprine；Meturedepa；Mitindomide；Meter Tuo Jinx；Mitocromin；Mitogillin；Mitomalcin；Mitomycin；Mitosper；Mitotane；Mitoxantrone hydrochloride；Wheat examines phenol Acid；Nafarelin；Nagrestipen；napavin；naphterpin；Nartograstim；Nedaplatin；Nemorubicin；Neridronic Acid； Nilutamide；Nysa mycin (nisamycin)；Nitric oxide modulator；Nitroxide antioxidant；Ni Tulin (nitrullyn)；Nocodazole；Nogalamycin；Ao Limosen (GENASENSE)；Octreotide；The gloomy ketone of fine jade (okicenone) difficult to understand； Olaparib (LYNPARZA)；Oligonucleotides；Onapristone；Ondansetron；Ao Luoxin (oracin)；Oral cytokine induction Agent；Ormaplatin；Oxisuran；oxaloplatin；Osaterone；Oxaliplatin；Losec Sa mycin (oxaunomycin)；Palau Mycin (palauamine)；Palmityl nitragin (palmitoylrhizoxin)；Pamidronic acid；Panaxytiol；Pa Nuo meter It is fragrant；Secondary coccus element；PARP (Poly ADP-ribose polymerase) inhibitor；Pazelliptine；Pegaspargase；Peldesine；Pentosan Polysulfate Sodium；Spray Si Tading；pentrozole；Perflubron；Perfosfamide；Perilla alcohol；Benzene azine mycin；Phenylacetate；Phosphate inhibits Agent；Sapylin；Pilocarpine hydrochloride；Pirarubicin；Piritrexim；Pula sting (placetin) A；Pula sting B；Methyl Mitomycin；Prednisone；Prostaglandin J2；Pyrazoloacridine；Taxol；PD035901；PD184352；PD318026； PD98059；Peliomycin；Pentamustine；Peplomycin sulfate；PKC412；Pipobroman；Piposulfan；Hydrochloric acid Piroxantrone；It is general Ka-7038Ⅶ；Plomestane；Podophyllotoxin；Polyphenol E；Porfimer Sodium；Methylmitomycin；Prednimustine；Procarbazine；Hydrochloric acid Procarbazine；Puromycin；Puromycin hydrochloride；Pirazofurin；Raltitrexed；Ramosetron；Demethyl retelliptine；Head mold Element；Rituximab；RII vitaminamide (retinamide)；Rogletimide；Rohitukine；Romurtide；Roquinimex； rubiginone B1；Such as Bo Xier (ruboxyl)；Riboprine；Romidepsin；rucaparib；Safingol；Hydrochloric acid is husky fragrant Dagger-axe；saintopin；Muscle phytol A；Sargramostim；Semustine；Sizofiran；Sobuzoxane: Sodium Borocaptate；Sodium phenylacetate；Rope It irrigates around (solverol)；Sonermin；Sorafenib；Sutent；Sparfosic acid；Racemomycin D；Spiromustine；Si Naipanding；Sea Continuous element 1；Spongistatin 2；Spongistatin 3；Spongistatin 4；Spongistatin 5；Spongistatin 6；Spongistatin 7；Spongistatin 8；With Spongistatin 9；Spiny dogfish Amine；This base of a fruit amide；Mesenchyma dissolves plain inhibitor；Sa Feinuoxin (sulfinosine)；suradista；Suramin；Bitter horse Legumin；SB239063；Department's beauty replaces Buddhist nun/AZD6244；Simtrazene；SP600125；Department's pool Suo Fei sodium；Sparsomycin；Hydrochloric acid germanium spiral shell Amine；Spiroplatin；Broneomycin；Streptozotocin；Sulofenur；Tallimustine；Tamoxifen methiodide；talazoparib(BMN 673)；Tauromustine；Tazarotene；Tecogalan sodium；Tegafur；For Lu Limu (tellurapyrylium)；For not mooring It is fragrant；Temozolomide；Teniposide；Ten oxide of tetrachloro；Tetrazolium amine (tetrazomine)；thaliblastine；Thiocoraline； Thrombopoietin；Thymalfasin；Thymopoietin receptor stimulating agent；Thymotrinan；Tirapazamine；Titanocene dichloride； Appropriate Pu Senting (topsentin)；Toremifene；Vitamin A acid；Triacetyluridine；Triciribine；Trimetrexate；Triptorelin；Support Alkane department fine jade；Turosteride；Tyrphostin；Tallysomycin；TAK-733；Taxotere；Tegafur；Hydrochloric acid replaces Lip river Anthraquinone；Teroxirone；Testolactone；Thiapurine；Thioguanine；Thiotepa；Tiazofurine；Tirapazamine；Citric acid Tuo Rui meter It is fragrant；Herceptin；Trestolone acetate；Phosphoric acid triciribine；Trimetrexate；Glucuronic acid Trimetrexate；Triptorelin；Salt Sour Tubulozole；TRAIL mRNA (TRAIL)；UBC inhibitor；Ubenimex；U0126；Crow Rameau department spit of fland；Uredepa；Vapreotide；Wei Ruierlin (variolin) B；Velaresol；Wei Lipani (ABT-888)；Black false hellebore Amine；Verteporfin；Vinorelbine；It ties up Sadinia (vinxaltine)；vitaxin；Vincaleukoblastinum；Vinblastine sulfate；Sulfuric acid Changchun is new Alkali；Eldisine；Vindesine sulfate；Sulfuric acid vinepidine；Sulfuric acid vinglycinate；Sulfuric acid vinleurosine；Tartaric acid Changchun is auspicious Shore；Sulfuric acid vinrosidine；Sulfuric acid vinzolidine；Vorozole；Wortmannin；XL518；Zanoterone；Zeniplatin；Zilascorb； Zinostatin stimalamer；Zinostatin；And zorubicin hydrochloride.

Other exemplary anticancer agents include Erbulozole (for example, R-55104)；Dolastatin 10 (for example, DLS-10 and NSC-376128)；Hydroxyl second sulphur mivobulin (for example, CI-980)；NSC-639829；Circle suberite lactone is (for example, NVP-XX- A-296)；ABT-751 (Abbott (Abbott)；For example, E-7010)；Atropic sea spit of fland A；Atropic sea spit of fland C；Hydrochloric acid Cemadotin (for example, LU-103793 and NSC-D-669356)；CEP 9722；Epothilones A；Epothilone B；Epothilone C；Angstrom slope is mould Plain D；Epothilone E；Epothilone F；Epothilone B N- oxide；Epothilones A N- oxide；16- azepine-angstrom slope is mould Plain B；21- aminoepothilone B；21- hydroxyepothilone D；26- fluoro Epothilones；The auspicious statin PE of Australia is (for example, NSC- 654663)；Rope benefit spit of fland (for example, TZT-1027)；LS-4559-P (Pharmacia Corp (Pharmacia)；For example, LS- 4577)；LS-4578 (Pharmacia Corp；For example, LS-477-P)；LS-4477 (Pharmacia Corp)；LS-4559 (method Ma West Asia company)；RPR-112378 (Aventis company (Aventis))；DZ-3358 (big needle company (Daiichi))；FR- 182877 (Teng Ze companies (Fujisawa)；For example, WS-9265B)；GS-164 (Wu Tian company (Takeda))；(the military field GS-198 Company)；KAR-2 (academy of sciences, Hungary (Hungarian Academy of Sciences))；BSF-223651(BASF；Example Such as, ILX-651 and LU-223651)；SAH-49960 (gift comes (Lilly)/Novartis (Novartis))；SDZ-268970 (gift/ Novartis)；AM-97 (A Maide (Armad)/consonance fermentation (Kyowa Hakko))；AM-132 (A Maide)；AM-138 (A Maide (Armad)/consonance fermentation (Kyowa Hakko))；IDN-5005 (Ying Aina (Indena))；Cryptophycin 52 (for example, LY-355703)；AC-7739 (aginomoto (Ajinomoto)；For example, AVE-8063A and CS-39.HCl)；AC-7700 (taste it Element；For example, AVE-8062；AVE-8062A；CS-39-L-Ser.HCl；And RPR-258062A)；Vitilevuamide；Micro-pipe Protein inhibitor A；Ghana's list rope；CA-170 (Kouris Co., Ltd (Curis))；Centaurcidin (for example, NSC-106969)；T- 138067(Tularik；For example, T-67；TL-138067 and TI-138067)；COBRA-1 (Parker Hughes research institute (Parker Hughes Institute)；For example, DDE-261 and WHI-261)；H10 (Kansas State University (Kansas State University))；H16 (Kansas State University)；Oncocidin A1 (for example, BTO-956 and DIME)；DDE-313 (pa Gram Hughes research institute)；Fijianolide B；Laulimalide；SPA-2 (Parker Hughes research institute)；SPA-1 (Parker Hughes Research institute；For example, SPIKET-P)；3-IAABU (cytoskeleton company (Cytoskeleton)/Mount Sinai School of Medicine (Mt.Sinai School of Medicine)；For example, MF-569)；Coscopin (for example, NSC-5366)；Nascapine；D- 24851 (Ace reaches drugmaker)；A-105972 (Abbott)；Hammett woods；3-BAABU (cytoskeleton company/west how mountain Medical college；For example, MF-191)；TMPN (Arizona State University (Arizona State University))；Dicyclopentadienyl second Acyl acetonation vanadium；T-138026(Tularik)；Monsatrol；Lnanocine (for example, NSC-698666)；3-IAABE is (thin Born of the same parents' skeleton company/Mount Sinai School of Medicine)；A-204197 (Abbott)；T-607(Tularik；For example, T-900607)；RPR- 115781 (Aventis company (Aventis))；Eleutherobin is (for example, demethylation Eleutherobin；Deacetylation Eleutherobin Element；Different Eleutherobin A；With Z- Eleutherobin)；Cali's shellfish glycosides；Cali's Belling；Halichondrin B；(Ace reaches system to D-64131 Medicine company)；D-68144 (Ace reaches drugmaker)；Diazonamide A；A-293620 (Abbott)；(the sea NPI-2350 Refreshing company (Nereus))；Spectromatic analysis；TUB-245 (Aventis company)；A-259754 (Abbott)； Diozostatin；(-)-phenylalanine (for example, NSCL-96F037)；D-62638 (Ace reaches drugmaker)；D-62636 (love Si Da drugmaker)；Myostromin B；D-43411(Zentaris；For example, D-81862)；A-289099 (Abbott)； A-318315 (Abbott)；HTI-286 is (for example, SPA-110；Trifluoroacetate) (Wyeth (Wyeth))；D-82317 (Zentaris)；D-82318(Zentaris)；SC-12983(NCI)；Li Sifu plast reaches spit of fland (Resverastatin) phosphoric acid Sodium；BPR-OY-007 (National Institutes of Health (National Health ResearchInstitutes))；And SSR- 250411 (matches Norfin, Inc (Sanofi))；Goserelin；Leuprorelin；Triptolide；Homoharringtonine；Topology is replaced Health；Itraconazole；Desoxyadenossine；Sertraline；Pitavastatin；Clofazimine；5- nonyl epoxide tryptamines；Wei Luofeini；Dabrafenib； Gefitinib (IRESSA)；Erlotinib (TARCEVA)；Cetuximab (ERBITUX)；Lapatinib (TYKERB)；Pa wood Monoclonal antibody (VECTIBIX)；Vande Thani (CAPRELSA)；Afatinib/BIBW2992；CI-1033/ Canertinib；Come that replace Buddhist nun/HKI-272；CP-724714；TAK-285；AST-1306；ARRY334543；ARRY-380；AG-1478；Up to can replace Buddhist nun/ PF299804；OSI-420/ demethyl Tarceva；AZD8931；AEE726；Pelitinib/EKB-569；CUDC-101； WZ8040；WZ4002；WZ3146；AG-490；XL647；PD153035；5- imuran；5- azepine -2'- deoxycytidine；17- N- allylamino -17-AAG (17-AAG)；- 1,25 dihydroxy vitamin d3 of 20- table；5 acetenyls urine is phonetic Pyridine；And BMS-599626.

One embodiment is for by combining with capecitabine and/or PLX4032 (Plexxikon company) and giving this hair The method of patient of the bright compound treatment with (optional is intractable) hepatocellular carcinoma.

It in another embodiment, is for giving the present invention by combining with capecitabine, Xeloda and/or CPT-11 Compound treatment (optional intractable) hepatocellular carcinoma method.

It in another embodiment, is for giving the present invention by combining with capecitabine, Xeloda and/or CPT-11 Compound treatment (optional intractable) hepatocellular carcinoma method.

It in another embodiment, is for by giving the compound of the present invention with capecitabine and irinotecan combination Patient of the treatment with (optional intractable) hepatocellular carcinoma or with can not cut off or the patient of metastatic hepatocellular carcinoma Method.

It in another embodiment, is for giving the compound of the present invention by combining with interferon-' alpha ' or capecitabine Treatment is with can not cut off or the method for the patient of metastatic hepatocellular carcinoma.

It in another embodiment, is for giving the compound of the present invention treatment with pancreas by combining with gemcitabine The method of the patient of gland cancer.

One embodiment is for giving chemical combination of the invention by combining with one or more medicament (or combinations thereof)s The method of patient of the object treatment with CCA.

One embodiment is for giving the compound of the present invention by combining with gemcitabine, cis-platinum (or combinations thereof) and controlling The method for treating the patient with CCA.

Pharmaceutical composition

" pharmaceutical composition " is preparation, which contains of the invention to the form of subject in being suitable for administration to Compound.In one embodiment, which is in bulk form or unit dosage forms.To be easy to give and realize dosage The dosage unit form compositions formulated of uniformity can be advantageous.The specification of dosage unit form by active agent uniqueness Characteristic and the particular treatment effect to be realized determine, and directly depend on them.

Possible preparation includes suitable for oral, sublingual, buccal, parenteral (for example, subcutaneous, intramuscular or intravenous), directly Intestines, part (including transdermal, intranasal and sucking) those of are given.For particular patient, most suitable mode of giving will be depended on The property and severity of positive treatment disease or the property of therapy used and the property for depending on reactive compound, but may If, oral give can be used for preventing and treating cancer.It can be provided in the form of discrete unit suitable for the oral preparation given, such as The respectively tablet of the reactive compound containing predetermined amount, capsule, cachet, pastille；It is provided with powder or particle；With aqueous or non- Solution or suspension in waterborne liquid provide；Or it is provided with oil-in-water or water-in-oil emulsion.

Include: pastille suitable for the sublingual or buccal preparation given, containing the compound of the present invention and typically seasons Matrix, the seasoning matrix such as sugar and Arabic gum or tragacanth gum；And pastille, it includes the active ingredients in inertial base Object, the inert substrate such as gelatin and glycerol or sucrose acacia.

Preparation suitable for parenterally giving typically comprises the aseptic aqueous solution of the reactive compound containing predetermined concentration； The solution can be isotonic with the blood of expected recipient.Other preparation suitable for parenterally giving includes containing physiology The preparation of upper suitable cosolvent and/or complexing agent (such as surfactant and cyclodextrin).For parenteral formulation, water packet Oil type emulsion is also suitable.Although these solution can be by intravenous administration, they can also pass through subcutaneous or flesh Interior injection is to give.

Preparation suitable for rectal administration may be provided as the suppository of unit dose, these suppositorys are included in and form suppository The compound of the present invention in one or more solid carriers (such as cocoa butter) of matrix.

Preparation suitable for part or intranasal administration includes ointment, emulsifiable paste, lotion, paste, gel, spray, aerosol And oil.Suitable carrier for such preparation includes vaseline, lanolin, polyethylene glycol, alcohol and combinations thereof.

Preparation of the invention can be prepared by any suitable method, typically via by the compound of the present invention with Liquid or solid carrier fine crushing or both mix uniformly and closely in a desired proportion, and then if desired, making institute It obtains mixture and is configured to desired shape.

For example, can be by suppressing the powder comprising active constituent or particle and one or more optional members (as bonded Agent, lubricant, inert diluent or surface active dispersing agent) homogeneous mixture, or by mold powdery active constituent Tablet is prepared with the immixture of inert liquid diluent.The suitable preparation given by sucking includes can be by various The particulate powders or mist that quantitative pressurised aerosol, sprayer or the insufflator of type generate.

The lung carried out via mouth is given, the partial size of powder or drop is typically at about 0.5 μM -10 μM or 1 μM of -5 μ In the range of M, to ensure to be delivered in bronchial tree.For intranasal administration, it can be used in about 10 μM of -500 μM of ranges Interior partial size, to ensure to be retained in nasal cavity.

Metered dose inhaler is the aerosol dispenser of pressurization, it typically contains the compound of the present invention in liquefied propulsion Suspension or solution formulation in agent.During use, preparation was penetrated engineered suitable delivering metered body by these devices The valve of product (typically from about 10 μM to 150 μM), it is spraying to generate the fine grained containing active constituent.Suitable propellant packet Include certain chlorofluorocarbon compounds, such as dicholorodifluoromethane, trichlorofluoromethane, dichlorotetra-fluoroethane and its mixture.It prepares Product can additionally contain one or more cosolvent, such as ethanol surfactant (such as three oleic acid of oleic acid or sorbitan Ester), antioxidant and suitable flavoring agent.

Sprayer is commercially available device, narrow by accelerating across compressed gas (typically air or oxygen) Venturi tube aperture or by ultrasonic agitation, the solution of active constituent or suspension are converted to therapeutic aerosols mist.With 40%w/w, simultaneously is up to about by be scattered in liquid-carrier and account for preparation in the suitable preparation used in sprayer And the active constituent that may include below about 20%w/w forms.The carrier is typically water or diluted water-alcoholic solutions, preferably It is allowed to body fluid be isotonic by adding such as sodium chloride.Optional additive includes preservative (if preparation is not sterile If preparation), such as methylparoban, antioxidant, flavoring agent, volatile oil, buffer and surfactant.

Preparation suitable for being given by insufflation includes fine powder pulverized powder, these powder can be blown by one kind Device delivers or brings into a manner of snuff nasal cavity.In insufflator, powder, which is comprised in, to be typically made of gelatin or plastics Capsule or cylindrantherae in, these capsules or cylindrantherae can be pierced or open in situ, and powder passes through device at the time of inhalation Or it is delivered by air that manual pump is pumped through.Powder for insufflator can be only made of or by comprising activity active constituent The powder blend of ingredient, suitable powder diluent (such as lactose) and optional surfactant forms.Active constituent is typical Ground account for preparation from about 0.1% to 100%w/w.

In another embodiment, the present invention provides pharmaceutical composition, which includes as active component The compound of the present invention is together at least one pharmaceutical carrier or diluent and/or the compound of the present invention and at least one drug The mixture of carrier or diluent.

Carrier is pharmaceutically acceptable and must be compatible with the other compositions in the composition, i.e., will not to other at Divide and adversely affects.Carrier can be solid or liquid, and be preferably formulated as unit dose preparation, such as may contain Have by weight from about 0.05% to 95% active constituent tablet.If desired, can also be by other physiologically active ingredients It is incorporated in pharmaceutical composition of the invention.

Other than the ingredient being particularly mentioned above, preparation of the invention may include the technical staff of pharmaceutical field Other medicaments known, this is related with the type of the preparation discussed.For example, being suitable for the oral preparation given may include adjusting Taste agent, and the preparation for being suitable for intranasal administration may include fragrance.

In one embodiment, give pharmaceutical composition with following dosage form, the dosage form include daily from about 0.1-1500mg, 0.2-1200mg、0.3-1000mg、0.4-800mg、0.5-600mg、0.6-500mg、0.7-400mg、0.8-300mg、1- The compound of the present invention of the total amount of 200mg, 1-100mg, 1-50mg, 1-30mg, 4-26mg, 5-25mg or 5-10mg.

The compound of the present invention can be applied in combination with other hepatocellular carcinoma therapeutic agents, such as anticancer chemotherapy medicine Object, hormone, one or more biological response modifiers and other angiogenesis inhibitors；Or with immunotherapy or gene therapy group It closes and uses.

The synthesis of 1. compound 1 of example

Compound 1 can be prepared by methods known in the art (for example, U.S. Patent number 7, described in 932,244 that A bit).For example, compound 1 can by as shown in scheme 1 and be disclosed in WO 2014/066819 method prepare.

Scheme 1

Step 1 is the esterification of compound 2 to obtain compound 4.Step 2 is that the reaction of compound 5 is formed from compound 4. Step 3 is protected in the hydroxyl group of the position C3 of compound 5 to obtain compound 6.Step 4 is the oxicracking of compound 6 To obtain compound 7.Step 5 is the reduction of compound 7 to obtain compound 8.Step 6 is the sulfonation of compound 8 to be changed Close the sodium salt (1-Na) of object 1.According to program known in the art, its free acid form can be converted (i.e. by the sodium salt of compound 1 Compound 1) or other salt forms (such as N of compound 1-DEA or compound 1, N- diethyl alkanamine).

The synthesis of 2. compound 2 of example

Compound 2 can by conventional method (for example, US publication 2009/0062526, U.S. Patent number 7,138, Those methods described in 390 and WO 2006/122977), as passed through the 6- step after generation compound 1 (shellfish cholic acid or OCA difficult to understand) Synthesis is to prepare, as shown in following scheme 2.

Scheme 2

The above method is a 6 steps synthesis.Step 1 is to use methanol esterification 7- in the presence of acidic catalyst and heating The C-24 carboxylic acid of keto lithcholic acid (KLCA), to generate methyl ester compound 7.Step 2 is to form silicon from compound a using highly basic Enol ether is then handled with chlorosilane to generate compound 8.Step 3 is the aldol condensation of the silylenolethers and acetaldehyde of compound 8 Reaction, to generate compound 10.Step 4 is the C-24 methyl esters saponification of compound 10, to generate compound 11.Step 5 is chemical combination The hydrogenation of the 6- ethylene moieties of object 11, to generate compound 12.Step 6 is the 7- keto group selective reduction of compound 12 At 7 Alpha-hydroxy groups, to generate compound 1.

Example 3.Mdr2^-/-And FXR^-/-The liver cancer of mouse occurs

Multi-drug resistance albumen 2 (Abcb4) is the member of ATP combination box (ABC) transport protein superfamily.Multi-drug resistance albumen 2 knock out mice (Mdr2^-/-) provide spontaneous hepatocellular carcinoma occur In vivo model (Katzenellengoben et al., Mol.Cancer Res. [molecule cancer research] 2007,5,11,1159-1170).What shortage was encoded by -2 gene of multi-drug resistance The mouse of Abc4 albumen develops into chronic surrounding inflammation and cholestatic liver disease, leads to the development of hepatocellular carcinoma.

Farnesoid X receptor albumen (FXR) is the nuclear receptor to work as the bile acid sensor of control bile acid stable state. This receptor is highly expressed in liver and other organs.FXR knocks out (FXR^-/-) develop liver cell gland after mouse is big at 15 months Tumor and cancer (Yang et al., Cancer Res. [cancer research], 2007,67,863).

OCA, compound 1-Na and control diet are had evaluated in Mdr2^-/-And FXR^-/-To the shadow of hepatocellular carcinoma development in mouse It rings.OCA is FXR agonist, and compound 1-Na is dual FXR/TGR5 agonist.Effect ratio OCA of the compound 1-Na to FXR It is about 10 times high.

Researching and designing

By Mdr2^-/-And FXR^-/-Mouse is randomly divided into three experimental groups.To mouse feed specific rodent diet or Feeding is supplemented with OCA, the diet of compound 1-Na or fed control diet 15 months.All mouse are under conditions of pathogen-free domestic In interior (23 DEG C) illumination in 12 hours/dark cycle raising of controlled temperature and random drinking water.Ethics Committee, Uni Degli Studi Di Bari (Ethical Committee of the University of Bari) has approved this experimental setup, which also obtains The approval of Italian Ministry of Health (Italian Ministry of Health), meets internationally recognized animal care guidelines. After 16 months, puts to death mouse and collect serum, liver and intestines.It counts the sum of liver tumour and measures the diameter of each tumour.

Processing group

Group 1: control diet

To mouse (n=4) fed control diet 15 months.

Group 2:OCA

It is supplemented with control diet 15 months of OCA with the dosage of 10mg/kg to mouse (n=8) feeding.

Group 3: compound 1-Na

It is supplemented with control diet 15 months of compound 1-Na with the dosage of 10mg/kg to mouse (n=15) feeding.

As a result

The Mdr2 of bile Salt treatment^-/-Liver inflammation and toxicity lead to the development of liver cell dysplasia in mouse.To 16 When month big, almost 100% Mdr2^-/-Control mice has all suffered from liver tumour.The FXR that age is 16 months^-/-Mouse suffers from certainly Hair property hepatocellular carcinoma.

Tumour reduces and size

Figure 1A and 2A shows compound 1-Na, OCA and control to Mdr2^-/-The influence of tumour quantity reduction in mouse.With it is right Photograph ratio, compound 1-Na clearly prevent the development of hepatocellular carcinoma in our current research.For compound 1-Na group, almost observe To significance,statistical (p=0.055), but as research used in control-animal (n=2) negligible amounts and be not implemented. Mdr2^-/-The mouse of control mice and OCA processing shows clear identifiable liver tumour, and finds in compound 1-Na group micro- Little tumour.Figure 1B and 2B shows the influence of compound 1-Na, OCA and control to the reduction percentage of diameter > 5mm tumour.In Mdr2^-/-In mouse, nearly 80% tumour found in OCA and control group has the diameter greater than 5mm.On the contrary, using compound The FXR of 1-Na, OCA and control treatment^-/-Mouse shows several big liver tumours, shows the development master for preventing hepatocellular carcinoma (Fig. 3 A, 3B and 4) if of FXR dependence.

Liver weight/weight (LW/BW)

Fig. 5 A and 5B describe the influence of compound 1-Na and control to the percentage of liver weight and weight.With first previous existence At data it is consistent, with compare and OCA processing Mdr2^-/-Mouse is compared, the Mdr2 of compound 1-Na processing^-/-Mouse shows The significant decrease of LW/BW ratio.In FXR^-/-LW/BW rate variance is not observed in group.

Biochemical parameter

In order to assess Mdr2^-/-And FXR^-/-The hepatic injury of mouse analyzes compound 1-Na and control to liver enzyme, alanine The influence of transaminase (ALT) and aspartate transaminase (AST) level.As shown in Figure 6 A and 6B, it is handled with compound 1-Na aobvious Writing reduces Mdr2^-/-ALT and AST in mouse is horizontal.However, in FXR^-/-ALT and AST water is not observed in the mouse of processing Flat difference.

Ileum FXR expression of target gene

In order to prove that FXR participates in prevention hepatocellular carcinoma, compound 1-Na and control are had evaluated to ileum FXR expression of target gene Influence.As expected, OCA and compound 1-Na are only in Mdr2^-/-Group moderate stimulation desmocyte growth factor-21 5 (Fgf15) and small heterodimeric partner (Shp) gene expression (Fig. 7 A and 7B).

Liver FXR expression of target gene

Cholesterol 7α-hydroxylase (cyp7a1) is the limit converted cholesterol in the classical biosynthesis pathway of bile acid Fast enzyme.Compound 1-Na and OCA are only in Mdr2^-/-Inhibit Cyp7a1 gene expression (Fig. 8) in mouse.Bile salt rear pump It (Bsep) is a kind of memebrane protein, using the energy of ATP hydrolysis come active transport bile salt.As illustrated in figures 9a and 9b, compound 1-Na gives induction Mdr2^-/-Liver Bsep activation in mouse.OCA does not promote liver Bsep to induce, this shows and compound 1-Na (FXR in its effective activation enteron aisle and liver) on the contrary, OCA in Mdr2^-/-It is less likely that there is liver activity in mouse.

Serum tolal bile acid

Compound 1-Na significantly reduces Mdr2^-/-Serum Ievel of total bile acids (Figure 10 A) in mouse.By observing FXR^-/- Do not occur to reduce in mouse and confirms the FXR dependence (Figure 10 B) of the discovery.

Different Effects of example 4.FXR or the TGR5 activation in cholangiocarcinoma progress

Method

By using 2 kinds of methods (mRNA microarray and qPCR) patient group (Copenhagen different with 2 kinds (Copenhagen) and Saint Sebastian (San Sebastian)) CCA people's biopsy and control in, and with (pass through qPCR) in the different people's CCA cell line of normal person's bile duct cell (NHC), determines FXR and TGR5 expression.Chronic Give specific FXR or TGR5 agonist (respectively OCA or compound 4；0.03% in food, continue 2 months；Intercept system Medicine company (Intercept Pharmaceuticals)) after, the model in situ of people CCA grows through in immunodeficient mouse Magnetic resonance imaging (MRI) assessment.FXR or TGR5 activation is also had evaluated to the proliferation of external CCA and NHC cell, migration and line grain The differentia influence of body energetic supersession.

As a result

Compared with the normal hepatocytes week in Copenhagen and Saint Sebastian group organizes with normal stones in intrahepatic bile duct, FXR is in people It is lowered in CCA tissue sample and TGR5 is raised (respectively by mRNA microarray and qPCR).The activation of FXR is increased by adjusting Grow, migrate and mitochondrial inhibit CCA progress (and the activation of TGR5 can promote the progress).To FXR and TGR5 activity Adjusting represent the potential treatment strategy of CCA.In vitro, compared with NHC, it was found that FXR is in different people's CCA cell line Lower reconciliation TGR5 up-regulation.

As shown in fig. s 11a through 11d, in CCA tumour FXR expression reduce and it is related to Tumor Differentiation: Figure 11 A represent and People around organizes (n=60) to compare, and the FXR mRNA microarray in the entire tissue (n=104) of CCA tumour expresses (Ge Benha Root group) (graceful-Whitney test (Mann-Whitney test))；Figure 11 B represents swollen according to the CCA of Tumor Differentiation hierarchal grouping FXR mRNA microarray expression in the entire tissue of tumor: (n=10) of well differentiated, (n=34) or the differentiation of moderate differentiation Undesirable (n=9) (Copenhagen group) (graceful-Whitney test)；Figure 11 C is represented and normal human liver tissue (n=20) and surrounding Human liver tissue (n=7) (Saint Sebastian group) is compared, and the FXR mRNA in CCA tumour (n=5) expresses (qPCR) (Man-favour Te Ni is examined)；And Figure 11 D represents the CCA tumour compared with matched surrounding human liver tissue (n=4) (Saint Sebastian group) In FXR mRNA express (qPCR) (paired-samples T-test).

As shown in fig. s 12a through 12d, in CCA tumour TGR5 expression increase, door week than in liver in CCA it is higher and (Figure 12 A) related to neural surrounding wetting.Figure 12 B is represented compared with people around organizes (n=60), the entire tissue of CCA tumour (n=104) the TGR5 mRNA microarray in expresses (Copenhagen group) (graceful-Whitney test).Figure 12 C is shown in clinical disease Parameter (anatomical position [in door week (n=36) or liver (n=68)] and neural surrounding wetting [negative (n=50) or the positive (n of science =42) TGR5 mRNA microarray expression (Copenhagen group) (graceful-Whitney inspection under]), in the entire tissue of CCA tumour It tests).Figure 12 C is represented compared with surrounding human liver tissue (n=27), and the TGR5 mRNA in CCA tumour (n=15) expresses (qPCR) (Saint Sebastian group) (graceful-Whitney test).Figure 12 D shows compared with matched surrounding human liver tissue (n=11), CCA (Wilcoxen matches Object Notation rank tests for TGR5 mRNA expression (qPCR) (Saint Sebastian group) in tumour (Wilcoxon matched-paires signed rank test))。

As illustrated in figures 13 a and 13b, compared with normal person's bile duct cell, FXR expression reduction and TGR5 in CCA cell line Expression increases: Figure 13 A shows the FXR mRNA in normal person's bile duct cell (n=6) and CCA cell line (respectively n=5 and 6) Express (qPCR) (graceful-Whitney test or unpaired T are examined)；Figure 13 B shows normal person's bile duct cell (n=6) and CCA cell It is the TGR5 mRNA expression (qPCR) in (respectively n=5 and 4) (non-matching T is examined or graceful-Whitney test).

Compared with untreated animal, the chronic FXR agonist OCA that gives of Xiang Yuanwei CCA mouse model inhibits tumour growth, And this influence is related to PCNA in the tumour for the animal treated and Ki67 expression reduction.As shown in Figure 14 A-14D, FXR Agonist Austria shellfish cholic acid (OCA) inhibits tumor growth in vivo, the expression reduction of this and proliferation mark, biliary tract mark and epithelium mark It is related.By EGI1 cell subcutaneous injection into CD1 nude mice male mice.Once neoplasm in situ is implanted into the naked hero of CD1 by tumour growth In the liver of property mouse.After two weeks, MRI analysis is carried out and starts treatment in the diet to give.It is monitored 1 month and 2 by MRI The tumour growth of the moon.Figure 14 A shows the representative for the mouse that untreated control mice, the mouse of OCA processing and compound 4 are handled Property MRI and liver image.The bar chart of Figure 14 B shows gross tumor volume multiple variation (the control n=8, OCA n quantitative by MRI =6, compound 4n=9 (graceful-Whitney test, single tail).Figure 14 C represent in the CCA tumour of liver original position proliferation mark (i.e. Ki67 and PCNA), the mRNA expression of bile mark (i.e. CK19) and epithelium mark (i.e. ZO-1).N=5-8 (do not match by graceful-Whitney T is examined, single tail).Figure 14 D shows the increasing in the liver original position CCA tumour of untreated or mouse with OCA or the processing of compound 4 Grow the representative immunohistochemistry image of mark (i.e. Ki67 and PCNA).On the contrary, being controlled with TGR5 agonist compound 4 is chronic The influence to CCA tumour growth is not observed in the animal for the treatment of.

Figure 17 shows the influence of OCA and compound 4 in the liver xenograft tumours expression of proliferation mark.Liver is in situ The mRNA expression of proliferation mark (i.e. Cdc25a, cyclin D1 and cyclinD3) in CCA tumour.N= 5-8 (graceful-Whitney or unpaired T are examined, single tail).

In vitro, inhibit the proliferation and migration of CCA cell, and the line that these events compare compared with OCA activation FXR It is related that mitochondrial energetics metabolism reduces (i.e. baseline breathing, maximum breathing and breathing relevant to ATP are reduced).On the other hand, FXR Activation does not influence the survival of CCA cell.As shown in figures 15 a-d, FXR agonist OCA inhibits CCA thin with dosage-dependent manner Born of the same parents are proliferated and inhibit CCA cell migration (it is related to the metabolism reduction of CCA cell Mitochondria), without inducing cell apoptosis.Figure 15A is shown compared with untreated control cell (n=4), handles the CCA cell of 48h (i.e. with 10 or 25 μM with OCA (n=5) EGI1 proliferation) (carries out) (non-matching T inspection) by flow cytometry using CFSE cell Proliferation dyeing.At least three It is measured in a independent experiment.Compared with untreated control cell, the CCA cell of 3-6-12h is handled at (25 μM) of OCA The mRNA expression of proliferation mark (i.e. Ki67, PCNA, cyclin D1 and cyclinD3) in (i.e. EGI1).N= 5-6 examines the unpaired T of untreated control.Figure 15 B shows the result of migration measurement in CCA cell (i.e. EGI1).Specified Time point and under the conditions of the wound healing measurement of (i.e. untreated control or OCA processing) and corresponding quantitative (in every kind of condition Under, n=6) representative micro-image (non-matching T inspection).It is measured in two independent experiments.Untreated and The representative micro-image of transwell migration room in the cell of OCA processing for 24 hours.Once measured.Figure 15 C, which is shown, to be made It stress test agent with the mitochondria in untreated or CCA cell (i.e. EGI1) with (25 μM) of OCA processing (pretreatment 3h) The hippocampus consumption rate (OCR) of box.The bar chart of the metabolizing parameters calculated in OCR measurement.Every group of n=11-12 (non-matching T inspection It tests).At least it is measured in three independent experiments.Figure 15 D represents the CCA of untreated or OCA processing with 10 or 25 μM In cell (i.e. EGI1), measured by the Apoptosis based on flow cytometry of annexin V and propidium iodide.It comes from The representative histogram of the merging data of two independent experiments and corresponding quantitative, total n=6-7 (non-matching T inspection).Solely at three It is measured in vertical experiment.

On the contrary, activate TGR5 with compound 4 stimulates the proliferation and migration of CCA cell really, and these events are compared with It is related that the mitochondrial of control increases (i.e. baseline breathing, proton leak and breathing relevant to ATP increase).Such as figure Shown in 16A-16C, proliferation, migration and the mitochondrial metabolism of 4 minimal irritation CCA cell of TGR5 agonist compound.Figure 16 A is shown Compared with untreated control cell, the proliferation for the CCA cell (i.e. EGI1) for handling 48h with compound 4 with 10 or 25 μM (passes through Flow cytometry is analyzed using CFSE cell Proliferation dyeing).N=5-6 (non-matching T inspection).In three independent experiments In be measured.Compared with untreated control cell, the CCA cell (i.e. EGI1) of 3-6-12h is handled at (25 μM) of compound 4 In proliferation mark (i.e. Ki67, PCNA, cyclin D1 and cyclinD3) mRNA expression.N=6 is not (to The unpaired T of processing control is examined or graceful-Whitney test).Figure 16 B represents the measurement of the migration in CCA cell (i.e. EGI1).In Specified time point and under the conditions of the wound healing measurement of (i.e. untreated control or compound 4 handle) and accordingly quantitative Representative micro-image.Merging data (total n=9 and 6) (non-matching T inspection) from two independent experiments.At least at three It is measured in independent experiment.The migration room transwell in the cell that untreated and compound 4 is handled for 24 hours and corresponding The representative micro-image of quantitative (n=4 and 2).It is measured in three independent experiments.Figure 16 C display uses untreated Or mitochondria in CCA cell (i.e. EGI1) with (25 μM) of compound 4 processing (pretreatment 3h) stress test kit Hippocampus consumption rate (OCR).The bar chart of the metabolizing parameters calculated in OCR measurement.Every group of n=11-12 (non-matching T inspection). At least it is measured in three independent experiments.

Claims (16)

1. a kind of method for the cancer for treating or preventing subject in need, this method include giving the FXR of therapeutically effective amount Agonist, the FXR agonist are selected from compound 1 or 2:

Or its pharmaceutically acceptable salt or amino acid conjugate.

2. the method as described in claim 1, wherein the cancer is selected from: hepatocellular carcinoma, cholangiocarcinoma, cancer of pancreas, kidney, prostate Cancer, the cancer of the esophagus, breast cancer, gastric cancer, kidney disease, salivary-gland carcinoma, oophoroma, carcinoma of uterine body, bladder cancer and lung cancer.

3. method according to claim 1 or 2, wherein the cancer is hepatocellular carcinoma.

4. method as claimed in claim 3, wherein the hepatocellular carcinoma is selected from the group, which is made up of: early hepatocyte Cancer, non-metastatic hepatocellular carcinoma, primary hepatoma, advanced hepatocellular carcinoma, Locally Advanced hepatocellular carcinoma, metastatic liver cell Cancer, paracmasis hepatocellular carcinoma or recurrent hepatocellular carcinoma after hepatectomy.

5. wherein the FXR agonist is compound 1 or it can pharmaceutically connect such as method of any of claims 1-4 The salt received.

6. method as claimed in claim 5, wherein the FXR agonist is the sodium salt of compound 1.

7. method as claimed in claim 5, wherein the FXR agonist is the N of compound 1, N- diethyl alkanamine.

8. wherein the FXR agonist is compound 2 or it can pharmaceutically connect such as method of any of claims 1-4 The salt or amino acid conjugate received.

9. method according to claim 8, wherein the FXR agonist is the glycine conjugates of compound 2.

10. method according to claim 8, wherein the FXR agonist is the taurine conjugate of compound 2.

11. method according to claim 8, wherein the FXR agonist is the sarcosine conjugate of compound 2.

12. a kind of pharmaceutical composition, which includes FXR agonist and pharmaceutically acceptable excipient, which swashs Dynamic agent is selected from the compound 1 or 2 for treating or preventing the cancer of subject in need:

Or its pharmaceutically acceptable salt or amino acid conjugate.

13. a kind of for treating or preventing the kit of the cancer of subject in need, which includes compound 1 or changes Close object 2:

Or its pharmaceutically acceptable salt or amino acid conjugate.

Purposes of the 14.FXR agonist in the drug for being used to prepare the cancer for treating or preventing subject in need, should FXR agonist is selected from compound 1 or 2:

Or its pharmaceutically acceptable salt or amino acid conjugate.

15. method according to claim 1 or 2, wherein the cancer is cholangiocarcinoma.

16. method as claimed in claim 15, wherein the FXR agonist is compound 1 or its pharmaceutically acceptable salt.