Combination of substituted 2, 4-diaminoquinoline compound and MEK inhibitor for treatment of liver cancer

Abstract

The present invention relates to combinations of substituted 2, 4-diaminoquinoline compounds and MEK inhibitors, pharmaceutical compositions containing them and their use as medicaments. The invention also relates to a combination of a substituted 2, 4-diaminoquinoline compound and a MEK inhibitor for use in the prevention and / or reduction and / or treatment of liver cancer. The substituted 2, 4-diaminoquinoline compound and the MEK inhibitor, which make up the combination, can be used simultaneously, independently or sequentially.

Description

Technical Field

[0001] The present invention relates to a combination of a substituted 2,4-diaminoquinoline compound and a MEK inhibitor, a pharmaceutical composition containing the same and its use as a medicament. The present invention also relates to a combination of a substituted 2,4-diaminoquinoline compound and a MEK inhibitor for preventing and / or reducing and / or treating liver cancer.

[0002] The substituted 2,4-diaminoquinoline compound and the mitogen-activated protein kinase kinase inhibitor (MEK inhibitor) constituting the combination may be used simultaneously, independently or sequentially. Background Art

[0003] KRAS mutations represent a driver event in pancreatic ductal adenocarcinoma (PDAC). To date, targeting non-wild-type KRAS in cancer therapy has proven challenging. However, targeting oncogene-driven signaling pathways represents a clinically validated approach in several cancers (e.g., chronic myeloid leukemia, melanoma). It has been described that a subpopulation of dormant tumor cells that survive KRAS ablation and contribute to tumor relapse are dependent on activation of mitochondrial function, lysosomal activity, and autophagy for survival. Furthermore, inhibition of the MEK→ERK signaling pathway results in PDAC cells being highly dependent on autophagy for survival. Mitogen-activated protein kinase kinase inhibitors (MEK inhibitors) force and drive cancer cells to be highly dependent on cellular autophagy for survival. Therefore, MEK inhibitors can induce autophagy dependency for cancer cell survival, which is a facile way for cancer cells to acquire resistance to MEK inhibitors in the oncology clinical setting.

[0004] Several studies have highlighted the link between the use of MEK inhibitors and effects on the autophagy pathway. PDAC has been found to be resistant to monotherapy with trametinib (a MEK inhibitor) or monotherapy with chloroquine / hydroxychloroquine (autophagy inhibitors), but is highly sensitive to dual therapy with trametinib + chloroquine at the preclinical level and to dual therapy with trametinib + hydroxychloroquine at the clinical level. Inhibition of the KRAS → RAF → MEK → ERK signaling pathway induces autophagy, which is responsible for cellular recycling, which protects PDAC from the cytotoxic effects of KRAS pathway inhibition (e.g., using MEK inhibitors). Inhibition of mitogen-activated protein kinase kinase 1 / 2 (MEK1 / 2 kinase) leads to activation of the LKB1 → AMPK → ULK1 signaling pathway, a key regulator of autophagy.

[0005] Induction of autophagy in response to KRAS inhibition was also observed with inhibition of the ERK effector downstream of KRAS. Similarly, inhibition of autophagy using autophagy inhibitors such as chloroquine or by using genetic or pharmacological autophagy inhibition was found to enhance the ability of ERK inhibitors to mediate an antitumor response in KRAS-driven pancreatic cells. Together, these data suggest that inhibition of the ERK signaling pathway drives cancer cells to become critically dependent on the autophagic process to acquire ERK resistance.

[0006] These studies are consistent with previous observations that autophagy acts as an adaptive and protective response to inhibition of KRAS → RAF → MEK → ERK signaling in cancer. Autophagy has also been implicated in resistance to a number of other standard anticancer chemotherapies. For example, autophagy induction has been found to be a cause of resistance in ovarian cancer to the highly cytotoxic agent paclitaxel. In the same manner as autophagy induction, ovarian and esophageal cancers can acquire the ability to escape cisplatin therapy. Lung cancer drug resistance to cisplatin can likewise be acquired through hypoxia-induced autophagy. In several other cancers, autophagy induction associated with the ER stress response leads to resistance to cyclin-dependent kinase inhibitors in primary patient chronic lymphocytic leukemia-derived cells and to HDAC inhibitors such as Tabastatin A in glioblastoma cell lines. Autophagy is particularly active during metabolic stress occurring in the tumor microenvironment and is a key survival pathway for cancer cells under other stress conditions, making autophagy a cellular partner of tumor growth.

[0007] Hepatocellular carcinoma (HCC) is an aggressive malignancy and the most common primary liver cancer characterized by highly aggressive biology and limited effective treatment options. An important signaling pathway in hepatocarcinogenesis is the MEK cascade involved in various cellular responses, including adaptation and survival. MEK plays a key role in this cascade, with MEK 1 / 2 representing a prototype and interesting therapeutic target for new oncology drugs. MEK 1 / 2 inhibitors represent a novel targeted therapy for the management of patients with advanced hepatocellular carcinoma (HCC). However, in some cancer models, inhibition of the KRAS → RAF → MEK → ERK signaling pathway induces autophagy, which is associated with tumor survival and selection of drug resistance. Therefore, incorporation of MEK inhibitors into HCC anticancer therapy may lead to a rapid loss of potent anticancer responses in the clinical setting through the acquisition of MEK inhibitor resistance. In the field of liver tumors, intrahepatic cholangiocarcinoma (iCCA) is the most common biliary malignancy. Surgical resection or liver transplantation is a potentially curative treatment option for patients with early-stage disease. However, most patients present with advanced disease and have limited treatment options. Acquired mutations of KRAS occur in approximately 20% of iCCAs. Activating mutations of KRAS lead to overactivation of the KRAS → RAF → MEK → ERK pathway, which increases cell proliferation and survival. MEK inhibition has shown efficacy in both wild-type and mutant KRAS (e.g., KRASV12D).

[0008] Therefore, there is a need to treat liver cancer, including hepatocellular carcinoma, hepatoblastoma, intrahepatic cholangiocarcinoma or extrahepatic cholangiocarcinoma, while maintaining a strong and durable response to anticancer therapy, particularly by avoiding the acquisition of resistance during MEK inhibitor therapy in the clinical setting. Summary of the invention

[0009] It has now been found that the combination of substituted 2,4-diaminoquinoline compounds and MEK inhibitors can achieve not only an overall additive effect but also a significant synergistic effect in inhibiting the growth of liver cancer cells, with potentially significant impact in vivo and clinical settings. These results open up a new avenue to overcome the potential MEK inhibitor resistance problem in liver anticancer therapy.

[0010] The present invention relates to a combination comprising:

[0011] ♦Compounds of formula (I)

[0012]

[0013] and pharmaceutically acceptable salts, hydrates, solvates, prodrugs, polymorphs, tautomers, isotopic variants, isomers, stereoisomers or mixtures of stereoisomers, and

[0014] ♦Mitogen-activated protein kinase kinase inhibitors (MEK inhibitors) and pharmaceutically acceptable salts, hydrates or solvates thereof.

[0015] The present invention also relates to a pharmaceutical composition containing such a combination, wherein each component of the combination may be contained in a separate pharmaceutical composition.

[0016] Another object of the present invention is a combination of a compound of formula (I) and a MEK inhibitor for use in the treatment and / or prevention and / or reduction of liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0017] The compound of formula (I) and the MEK inhibitor or the pharmaceutical composition containing them can be administered simultaneously, independently or sequentially.

[0018] The present invention also relates to a kit comprising a combination of a compound of formula (I) and a MEK inhibitor, wherein both or either of the components are in the form of a pharmaceutical composition that can be administered simultaneously, separately or sequentially. DETAILED DESCRIPTION

[0019] The present invention relates to a combination comprising

[0020] ♦Compounds of formula (I)

[0021]

[0022] in

[0023] • L 1 Selected from a single bond, an optionally substituted (-CH 2 -) p group, optionally substituted alkylene, carbonyl;

[0024] • R 1 is selected from the group consisting of hydrogen atom, halogen atom, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted alkoxy, -OR 7 、-O-(CO)-R 7 、-O-(CO)-NR 5 R 6 、-NR 5 -(CO)-R 7 、-O-(CO)-OR 7 、-NR 5 -(CO)-OR 7 , azido, hydroxy, cyano, nitro, -NR 5 R 6;

[0025] • R 2 and R 3 Simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted heterocyclic group, an optionally substituted aryl group, an optionally substituted benzyl group, an optionally substituted heteroaryl group;

[0026] or R 2 and R 3 may be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group;

[0027] • R 4 is selected from the group consisting of hydrogen atom, halogen atom, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted alkoxy, -OR 7 、-O-(CO)-R 7 、-O-(CO)-NR 5 R 6 、-NR 5 -(CO)-R 7 、-O-(CO)-OR 7 、-NR 5 -(CO)-OR 7 , azido, hydroxy, cyano, nitro, -NR 5 R 6 ;

[0028] • R 5 and R 6 are simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted heterocyclic group, an optionally substituted aryl group, an optionally substituted heteroaryl group;

[0029] or R 5 and R 6 may be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group;

[0030] • R 7 may be selected from optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl;

[0031] • n is an integer which can have any value of 0, 1, 2, 3 or 4;

[0032] • m is an integer which can have any value of 0, 1, 2, 3, 4 or 5;

[0033] • p is an integer which can have any value of 0 or 1;

[0034] and pharmaceutically acceptable salts, hydrates, solvates, prodrugs, polymorphs, tautomers, isotopic variants, isomers, stereoisomers or mixtures of stereoisomers, and

[0035] ♦Mitogen-activated protein kinase kinase inhibitors (MEK inhibitors) and pharmaceutically acceptable salts, hydrates or solvates thereof.

[0036] ♦The term "bond" or "single bond" refers to a chemical bond between two atoms or two moieties (when the atoms connected by the bond are considered to be part of a larger substructure).

[0037] ♦ The expression "simultaneously or independently" is used herein to indicate that the variable applies in either case, regardless of whether there are variables with the same or different definitions in the same compound. Thus, for example, in a compound where a substituent Xi occurs twice and is defined as "simultaneously or independently a group G1 or a group G2", both Xi can be G1 (simultaneously), both Xi can be G2 (simultaneously), or one Xi can be G1 and the other Xi can be G2 (independently).

[0038] ♦ The expressions "halogen", "halogen atom" wherever they appear mean one or more atoms which may be selected from fluorine (F), chlorine (Cl), bromine (Br), iodine (I), preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine.

[0039] ♦The term "alkyl", alone or in combination with other groups, refers to a group of 1 to 20 carbon atoms (C 1 -C 20 carbon atoms), preferably 1 to 16 carbon atoms (C 1 -C 16 carbon atoms), more preferably 1 to 10 carbon atoms (C 1 -C 10 The alkyl group may be optionally substituted as defined herein.

[0040] ♦ The term "alkylene" refers to an alkyl group as described above, wherein the alkyl group is a divalent group, also defined as an alkanediyl group. Typically, an alkylene group has two points of attachment to the rest of the molecule (e.g., -L in a molecule represented by formula (I) of the present invention). 1-). The two points of attachment of an alkylene group may be located on one specific carbon atom or on two different carbon atoms thereof.

[0041] ♦ The term "lower alkyl", alone or in combination, means a straight or branched chain alkyl group having 1 to 10 carbon atoms ("C 1 -C 10 -alkyl"), preferably a straight chain or branched chain alkyl group having 1 to 5 carbon atoms ("C 1 -C 5 -alkyl"), particularly preferably a straight-chain or branched alkyl group having 1 to 3 carbon atoms ("C 1 -C 3 -alkyl"). Lower alkyl may be optionally substituted as defined herein. Non-limiting examples of straight chain and branched lower alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, isomeric pentyls, isomeric hexyls, isomeric heptyls, isomeric octyls, isomeric nonyls, isomeric decyls, preferably methyl and ethyl and n-propyl and isopropyl and tert-butyl and isobutyl and sec-butyl and isomeric pentyls, most preferably methyl and ethyl and n-propyl and isopropyl and n-butyl and tert-butyl.

[0042] ♦The term "alkenyl" means a group containing an olefinic bond. A straight-chain or branched hydrocarbon residue, wherein R', R'', R''' and R IV refers to the remaining parts of the alkenyl group, which may be the same or different. R', R'', R''' and R IV The alkenyl group may have 2 to 10 carbon atoms ("C 2 -C 10 -alkenyl"), preferably 2 to 5 carbon atoms ("C 2 -C 5 -alkenyl"), particularly preferably 2 to 4 carbon atoms ("C 2 -C 4 The alkenyl moiety may be branched, straight chain or cyclic (in which case it is also referred to as a "cycloalkenyl"). The alkenyl group may also be a "lower alkenyl" having 2 to 6 carbon atoms, including all isomeric forms (cis, trans, Z, E). The alkenyl group may be optionally substituted as defined herein. Non-limiting examples of lower alkenyl groups are ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, isobutenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 3-penten-2-yl, 3-penten-4-yl, isopentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, isohexenyl. Preferred examples are ethenyl, 1-propenyl, 2-propenyl, 2-butenyl, 2-buten-2-yl and isopentenyl.

[0043] ♦The term "alkynyl" means a straight or branched hydrocarbon residue containing an acetylenic bond, wherein two carbon atoms form a triple bond R'―C≡C―R", wherein R' and R" refer to the rest of the alkynyl group and they may be the same or different. The R' and R" portions of the alkynyl moiety may be branched, straight or cyclic. Alkynyl groups may have from 2 to 10 carbon atoms ("C 2 -C 10 -alkynyl"), preferably 2 to 5 carbon atoms ("C 2 -C 5 -alkynyl"), particularly preferably 2 to 4 carbon atoms ("C 2 -C 4 -alkynyl”). Alkynyl can be optionally substituted as defined herein. Non-limiting examples of alkynyl are ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 3-butynyl, 4-butynyl, but-2-yn-1-yl, 1-pentynyl, pent-2-yn-1-yl, pent-3-yn-1-yl, pent-4-yn-1-yl, pent-2-yn-3-yl. Preferred examples are propyn-1-yl, propyn-3-yl, butyn-1-yl, butyn-3-yl, butyn-4-yl, but-2-yn-1-yl. Alkynyl can also be a “lower alkynyl” (“C ynyl”) having 2 to 6 carbon atoms. 2 -C 6 -alkynyl").

[0044] ♦The term "cycloalkyl" means a monocyclic or polycyclic group containing only carbon and hydrogen, which is a saturated ring and contains 3 to 12 carbon atoms ("C 3 -C 12 -cycloalkyl"), for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecane. Cycloalkyl includes radicals having 3 to 12 ring atoms ("C 3 -C 12 -cycloalkyl"), preferably 3 to 8 ring atoms ("C 3 -C 8 -cycloalkyl”), more preferably 3 to 7 ring atoms (“C 3 -C 7 -cycloalkyl"), more preferably 3 to 6 ring atoms ("C 3 -C 6 -cycloalkyl") radical. Depending on the structure, the cycloalkyl may contain adjacent and substituted cycloalkenyl and / or alkenyl radicals. The cycloalkyl may be optionally substituted as defined herein.

[0045] ♦The term "cycloalkenyl" means a non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond C=C, comprising about 3 to about 12 carbon atoms, preferably about 5 to about 10 carbon atoms, more preferably about 5 to about 7 carbon atoms, more preferably about 5 to about 6 carbon atoms. The cycloalkenyl group may be optionally substituted with one or more "ring system substituents", which may be the same or different and are as defined above. Non-limiting examples of monocyclic cycloalkenyl groups include cyclopentenyl, cyclohexenyl, cyclohepta-1,3-dienyl, and the like. Non-limiting examples of suitable polycyclic cycloalkenyl groups are, for example, norbornenyl, and the like. The cycloalkenyl group may be optionally substituted as defined herein.

[0046] ♦The term "cycloalkynyl" means a non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon triple bond (C≡C) comprising about 8 to about 12 carbon atoms, preferably about 8 to about 10 carbon atoms. The cycloalkynyl group may be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined above. Non-limiting examples of monocyclic cycloalkynyl groups include cyclooctynyl, cyclononynyl, cyclodecynyl, and the like. The cycloalkynyl group may be optionally substituted as defined herein.

[0047] ♦ The terms "haloalkyl", "haloalkenyl", "haloalkynyl" and "haloalkoxy" include alkyl, alkenyl, alkynyl and alkoxy structures as defined herein, wherein at least one hydrogen is replaced with a halogen atom as defined herein. In certain embodiments where two or more hydrogen atoms are replaced with halogen atoms, the halogen atoms are identical to one another. In other embodiments where two or more hydrogen atoms are replaced with halogen atoms, the halogen atoms are identical to one another or are not identical to one another.

[0048] ♦The term "fluoroalkyl" as used herein refers to a straight or branched chain alkyl group as defined herein, wherein at least one hydrogen atom is replaced by a fluorine atom. Examples of fluoroalkyl groups include, but are not limited to -CH 2 F, –CHF 2 , -CF 3 、-CH 2 CF 3 , -CF 2 CF 3 , -CF 2 CH 3 、-CH 2 CH 2 CF 3 、-CH(CF 3 ) 2 , -CF 2 CH(CH 3 ) 2 Etc. "Fluoroalkyl" may be optionally substituted as defined herein.

[0049] ♦“Alkoxy” refers to -O(C 1 -C 10 -O(alkyl) group, -O(cycloalkyl) group, -O(heterocyclyl) group, wherein "C 1 -C 10 Alkyl", "C 1 -C 10 "Cycloalkyl" and "heterocyclyl" are as defined herein. The term "lower alkoxy" refers to the group R'-O-, wherein R' is a lower alkyl group and the term "lower alkyl" has the meaning given previously. Alkoxy may be optionally substituted as defined herein. Non-limiting examples of lower alkoxy are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy, n-pentoxy, n-hexoxy, preferably methoxy and ethoxy and isopropoxy and tert-butoxy, most preferably methoxy and ethoxy. Non-limiting examples of "alkoxy" having an -O(cycloalkyl) group are cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy.

[0050] ♦The term "heterocyclyl", "non-aromatic heterocycle", "heterocycloalkyl", "heterocyclic group" or "heteroalicyclic group" refers to a fully saturated or unsaturated but not fully unsaturated, 3- to 9-membered monocyclic group, preferably a 3- to 7-membered monocyclic group, more preferably a 3- to 6-membered monocyclic group, or a fused heterocyclic ring system containing 5 to 16 atoms, preferably 5 to 14 atoms, more preferably 5 to 10 atoms, more preferably 5 to 9 atoms, which has at least one heteroatom, and the heteroatom when more than one is simultaneously or independently selected from an oxygen atom, a nitrogen atom or a sulfur atom. Each ring of the heterocyclyl may have at least one heteroatom, and the heteroatom when more than one is simultaneously or independently selected from a nitrogen atom, an oxygen atom and / or a sulfur atom. "Heterocycloalkyl" may be optionally substituted as defined herein. The covalent bond to "heterocycloalkyl" may be at a heteroatom or through a carbon atom. In certain embodiments, the non-aromatic heterocycle contains one or more carbonyl or thiocarbonyl groups, such as oxo-containing groups and thio-containing groups. The heterocyclic group may be optionally substituted as defined herein. Examples of heterocycloalkyl groups include, but are not limited to, lactams, lactones, cyclic imides, cyclic thioimides, cyclic carbamates, cyclic ureas, tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-dihydropyran ... oxane, 1,3-dioxane, 1,4-dioxane oxadiazine, 1,4-dioxane, piperazine, 1,3-oxathiazine, 1,4-oxathiazine, 1,4-oxathiazine, tetrahydro-1,4-thiazine, 2H-1,2- Azine, maleimide, succinimide, barbituric acid, thiobarbituric acid, dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane, hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran, pyrroline, pyrrolidine, pyrrolidone, pyrrolidinedione, pyrazoline, pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole, 1,3-dioxolane, 1,3-dithiole, 1,3-dithiolane, isocyanate, Oxazoline, Iso Oxazolidine, Oxazoline, Oxazolidine, Oxazolidinone, thiazoline, thiazolidine, 1,3-oxathiolane and their N-oxides (for amino heterocycles).

[0051] ♦The term "aryl" refers to an aromatic ring in which each atom forming the ring is a carbon atom. The aryl ring may contain five, six, seven, eight, nine or more than nine carbon atoms, preferably 5 to 16 carbon atoms, more preferably 5 to 12 carbon atoms, more preferably 5 to 10 carbon atoms, and means any stable monocyclic, bicyclic and tricyclic ring system in which at least one ring is aromatic. Aryl groups may be optionally substituted as defined herein. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, phenanthrenyl, anthracenyl, fluorenyl, biphenyl and indenyl. Depending on the structure, an aryl group may be a monovalent group or a divalent group (in which case it is referred to as an arylene group). Examples of arylene groups include, but are not limited to, benzene-1,2-diyl, benzene-1,3-diyl, benzene-1,4-diyl, naphthalene-2,7-diyl, naphthalene-2,6-diyl, naphthalene-1,4-diyl, naphthalene-1,5-diyl, acenaphthene-diyl, phenanthrene-3,8-diyl, fluoranthene-diyl, 3-methylbenzene-1,4-diyl, and the like.

[0052] ♦ The term "heteroaryl" generally refers to an aromatic 5- or 11-membered ring which contains at least one heteroatom and may additionally contain one, two, three or four atoms selected from nitrogen, oxygen and / or sulfur, for example pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, 2-oxo-1,2-dihydropyridinyl, Oxazolyl, iso oxazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, furanyl, The term "heteroaryl" also refers to a bicyclic aromatic or partially unsaturated radical comprising two 5-membered or 6-membered rings, one or both of which may contain one, two, three or four atoms selected from nitrogen, oxygen or sulfur, such as quinolyl, isoquinolyl, cinnolinyl, pyrazolyl, imidazolyl, thiazolyl, thienyl, furanyl, oxazolyl, isothiazolyl, pyrazolo[1,5-a]pyridinyl, imidazo[1,2-a]pyridinyl, quinoxalinyl, quinazolinyl, benzothiazolyl, benzotriazolyl, 1H-benzo[d]imidazole, benzo[d]isothiazolyl, oxazolyl, benzo[d]isothiazolyl, benzo[c]isothiazolyl oxazolyl, benzo[c]isothiazolyl, indolyl, isoindolyl, 6,7-dihydro-5H-pyrrolo[3,4-b]pyridinyl, 2,3-dihydro-1H-pyrrolo[3,4-c]pyridinyl, 6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidinyl, purinyl, indazolyl, indolizinyl, imidazo[1,2-a]pyridinyl, imidazo[1,5-a]pyridinyl, imidazo[1,5-a]pyrazinyl, imidazo[1,2-a]pyrazinyl, 1H-imidazo[4,5-b]pyrazinyl, pyrazolo[1,5-a]pyridinyl, pyrrolo[1,2-a]pyrimidinyl, pyrrolo[1,2-a]pyrazinyl, pyrrolo[1,2-c]pyrimidinyl, Azo[4,5-b]pyridinyl, Azo[4,5-c]pyridinyl, Azo[5,4-c]pyridinyl, oxazolo[5,4-b]pyridinyl, thiazolo[4,5-b]pyridinyl, thiazolo[4,5-c]pyridinyl, thiazolo[5,4-c]pyridinyl, thiazolo[5,4-b]pyridinyl, Azo[5,4-d]pyrimidinyl, oxazolo[4,5-d]pyrimidinyl, thiazolo[5,4-d]pyrimidinyl, thiazolo[4,5-d]pyrimidinyl, oxazolo[4,5-b]pyrazinyl, thiazolo[4,5-b]pyrazinyl, iso oxazolo[4,5-b]pyrazinyl, isothiazolo[4,5-b]pyrazinyl, isothiazolo[4,5-b]pyrazinyl oxazolo[4,5-d]pyrimidinyl, isothiazolo[4,5-d]pyrimidinyl, isothiazolo[4,5-d]pyrimidinyl oxazolo[5,4-d]pyrimidinyl, isothiazolo[5,4-d]pyrimidinyl, isothiazolo[5,4-d]pyrimidinyl oxazolo[5,4-b]pyridinyl, isothiazolo[5,4-c]pyrimidinyl, isothiazolo[5,4-c]pyrimidinyl oxazolo[5,4-c]pyridinyl, isothiazolo[4,5-c]pyridinyl, isothiazolo[4,5-c]pyridinyl Azo[4,5-c]pyridinyl, iso Azo[4,5-b]pyridinyl, iso oxazolo[4,3-d]pyrimidinyl, isothiazolo[4,3-d]pyrimidinyl, isothiazolo[4,3-d]pyrimidinyl oxazolo[3,4-d]pyrimidinyl, isothiazolo[3,4-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, pyrido[3,2-d]pyrimidinyl, pyrido[2,3-b]pyrazinyl, pyrido[3,4-b]pyrazinyl, [1,2,3]triazolo[4,5-b]pyridinyl, [1,2,3]triazolo[4,5-c]pyridinyl, 3H-[1,2,3]triazolo[4,5-d]pyrimidinyl. Preferred heteroaryl groups are pyridinyl, pyrazinyl, pyrimidinyl, thiazolyl, isothiazolyl, Azolyl, iso oxazolyl, quinazolinyl and pyrazinyl.

[0053] ♦ The term "heteroaryl" or "heteroaromatic" refers to an aromatic group comprising one or more rings and one or more heteroatoms independently selected from nitrogen (N), oxygen (O) and sulfur (S). An N-containing "heteroaryl" or "heteroaromatic" moiety refers to an aromatic group in which at least one skeletal atom of the ring is a nitrogen atom (N). Heteroaryl groups may be optionally substituted as defined herein.

[0054] ♦The term "heteroaryl" or "heteroaromatic" also refers to a fused heteroaryl system in which two or more rings share one or more bonds, comprising 7 to 16 ring atoms, preferably 8 to 13 ring atoms, more preferably 8 to 10 ring atoms, comprising 1, 2, 3, 4 or 5 heteroatoms selected simultaneously or independently from nitrogen (N), oxygen (O) and sulfur (S), and comprising at least one carbon atom (C), provided that the fused ring does not contain adjacent oxygen (O) and / or sulfur (S) atoms. The "heteroaryl" may be optionally substituted as defined herein. The covalent bond to the heteroaryl may be at a heteroatom or through a carbon atom. Also included are N-oxides of ring nitrogen, and heteroaryl groups in which the ring nitrogen is substituted by an optionally substituted alkyl group to form a quaternary amine. Preferred heteroaryl groups are pyridyl, pyrimidinyl, thiazolyl, isothiazolyl, Azolyl, iso The heteroaryl groups are substituted with oxazolyl, quinazolinyl, pyrazinyl and N-oxides thereof. All positional isomers are contemplated (e.g., pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyridin-5-yl, pyridin-6-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl and pyrimidin-6-yl). The heteroaryl groups may be optionally substituted as defined herein.

[0055] ♦ The expression "optionally substituted" means unsubstituted or substituted simultaneously or independently by one or more substituents selected from the group consisting of halogen atoms, hydroxyl, cyano, azido, -nitro, carboxyl, -CF 3, alkyl as defined herein, haloalkyl as defined herein, fluoroalkyl as defined herein, alkenyl as defined herein, alkynyl as defined herein, cycloalkyl as defined herein, cycloalkenyl as defined herein, cycloalkynyl as defined herein, heterocyclyl as defined herein, alkoxy as defined herein.

[0056] ♦The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of a free base or free acid, which is not undesirable biologically or otherwise. The salt is formed using an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., preferably hydrochloric acid, and an organic acid such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, salicylic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, glutaric acid, cinnamic acid, mandelic acid, malic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, N-acetylcysteine, N-acetyllysine, lysine and N-acetylarginine, arginine, etc. In addition, these salts can be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium salts, etc. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyamine resins, etc. The compounds of formula (I), (II), (III) or (IV) may also exist in the form of zwitterions.

[0057] ♦ A particularly preferred pharmaceutically acceptable salt of the compound of formula (I), (II), (III) or (IV) is the hydrochloride salt.

[0058] The compounds of formula (I), (II), (III) or (IV) may also be solvated, for example hydrated. The solvation may be carried out during the manufacturing process or may occur, for example, as a result of the hygroscopic properties of the initially anhydrous compound of formula (I) (hydration). The term "pharmaceutically acceptable salt" also includes physiologically acceptable solvates, including water.

[0059] ♦ The mitogen-activated protein kinase kinase inhibitors (MEK inhibitors) described herein may also be solvated, for example hydrated. The solvation may occur during the manufacturing process (e.g. dimethyl sulfoxide solvate form) or may occur as a result of the hygroscopic properties of the initially anhydrous MEK inhibitor (e.g. hydration of the hydrated form). The term "pharmaceutically acceptable salt" also includes physiologically acceptable solvates, including water.

[0060] ♦ "Isomers" are compounds that have the same molecular formula but differ in the nature or order of bonding of their atoms or in the arrangement of their atoms in space. Isomers that differ in the arrangement of their atoms in space are called "stereoisomers". Stereoisomers that are not mirror images of each other are called "diastereomers", and those that are non-superimposable mirror images are called "enantiomers", sometimes also called optical isomers.

[0061] ♦ "Prodrug" means a compound that undergoes transformation within a biological system to become a compound of the invention. A prodrug is a chemical derivative that is inactive or less active than the drug itself. After administration and diffusion in vivo, the prodrug derivative undergoes one or more metabolic processes to release the active drug. The conversion of the prodrug to the drug is usually carried out under the control of enzymatic processes (usually by metabolic means such as hydrolysis, reduction or oxidation), and less often by classical chemical reactions during its diffusion in vivo. The connection between the carrier and the drug can be, but is not limited to, esters, amides, carbonates, carbamates, imines, acetals, ethers (e.g., glucuronic acid coupling), oxidizable functions and molecular systems, reducible functions and reducible molecular systems, photoactivatable functions and photoactivatable molecular systems. For example, an ester prodrug of a compound containing a hydroxyl group may be convertible to the parent molecule by hydrolysis in vivo. Suitable esters of compounds of the invention containing hydroxy groups are, for example, acetates, citrates, lactates, tartrates, malonates, oxalates, salicylates, propionates, succinates, fumarates, maleates, methylene bis-β-hydroxynaphthoate, gentisates, isethionates, di-p-tolyl tartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, cyclohexylaminosulfonates and quinates. As another example, ester prodrugs of compounds of the invention containing carboxyl groups may be converted to the parent molecule by in vivo hydrolysis (examples of ester prodrugs are described by FJ Leinweber, Drug Metab. Res. 1987, (18) pp379, incorporated herein by reference). Similarly, acyl prodrugs of compounds containing amino groups may be convertible to the parent molecule by in vivo hydrolysis (examples of prodrugs of these and other functional groups (including amines, alcohols) are described in Prodrugs: Challenges and Rewards (Part 1 and Part 2), V. Stella, R. Borchardt et al., eds., Springer, 2007, and Prodrugs and Targeted Delivery: Towards Better ADME Properties, J. Rautio, Seies Ed. R. Mannhold, H. Kubinyl, G. Folkers, eds., Wiley-VCH 2011, each of which is incorporated herein by reference).

[0062] ♦ Prodrug carrier systems are often used to increase aqueous or lipid solubility, reduce toxicity, improve chemical and biological stability of sensitive compounds, and increase in vivo circulation time (T 1 / 2), increasing total drug exposure (AUC) and organ distribution (PK-PD analysis) as well as site-specific targeting.

[0063] ♦The term "pharmaceutically acceptable carrier" is intended to include any and all materials compatible with drug administration, including solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and other materials and compounds compatible with drug administration. Unless any conventional media or agents are incompatible with the active compound, their use in the compositions of the present invention will be considered. Supplementary active compounds may also be incorporated into the compositions. These compositions can be prepared by applying techniques known in the art, such as those described in Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems (10th Edition), 2014, edited by Loyd Allen, Howard C. Ansel, published by Wolters Kluwer Health, and Remington: The Science and Pratice of Pharmacy (22nd Edition), 2012, edited by Loyd V. Allen, published by Pharmaceutical Press.

[0064] ♦ As used herein, the terms "subject" or "patient" are used interchangeably. As used herein, the term "subject" refers to animals (e.g., birds, reptiles, and mammals), preferably mammals, including non-primates (e.g., camels, donkeys, zebras, cows, pigs, horses, goats, sheep, cats, dogs, rats, and mice) and primates (e.g., monkeys, chimpanzees, and humans), most preferably humans.

[0065] ♦The term "therapy" as used herein may refer to any regimen, method, composition, formulation and / or agent that can be used to prevent, treat, manage or improve a disease. In certain embodiments, the term "therapy" refers to biological therapy, supportive therapy and / or other therapies known to those skilled in the art that can be used to treat, manage, prevent or improve different diseases.

[0066] ♦The term "cholangiocarcinoma" refers to cancer that forms in the bile ducts, which are a series of thin tubes that run from the liver to the small intestine. The bile ducts carry a fluid called bile from the liver and gallbladder to the small intestine, where it helps digest fats in food. Cholangiocarcinoma is also called bile duct cancer.

[0067] ♦The term "intrahepatic cholangiocarcinoma" refers to bile duct cancer that occurs in the part of the bile ducts inside the liver.

[0068] ♦The term "extrahepatic cholangiocarcinoma" refers to bile duct cancer that occurs in the part of the bile duct closest to the small intestine.

[0069] ♦The term "therapeutically effective amount" of a compound refers to an amount of the compound that is effective in preventing, alleviating or ameliorating symptoms of a disease or prolonging the survival of the subject being treated. Determination of a therapeutically effective amount is within the skill of the art. The therapeutically effective amount or dosage of a compound according to the invention may vary within a wide range and may be determined in a manner known in the art. Such dosage will be adjusted according to the individual requirements in each particular case, including the specific compound being administered, the route of administration, the condition being treated, and the patient being treated. In general, in the case of oral or parenteral administration to an adult weighing about 70 kg, the daily dose is:

[0070] - for a compound of formula (I), (II), (III) or (IV) or a pharmaceutically acceptable salt or solvate thereof, 50 mg to 500 mg, preferably 50 mg to 200 mg, more preferably 50 mg to 100 mg,

[0071] and

[0072] - for a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof, 0.5 mg to 150 mg, preferably 1 mg to 100 mg, more preferably 1.5 mg to 90 mg should be suitable,

[0073] Although the upper limits may be exceeded where indicated,

[0074] Or for the MEK inhibitor trametinib or a pharmaceutically acceptable salt or solvate thereof, 0.5 mg to 5 mg, preferably 0.5 mg to 2 mg, more preferably 1 mg to 2 mg, more preferably 1.5 mg to 2 mg should be suitable, although the upper limit may be exceeded when indicated.

[0075] Or for the MEK inhibitor cobimetinib or a pharmaceutically acceptable salt (e.g. fumarate) or solvate thereof, 10 mg to 80 mg, preferably 10 mg to 60 mg, more preferably 20 mg to 60 mg, more preferably 40 mg to 60 mg should be suitable, although the upper limit may be exceeded when indicated.

[0076] The daily dosage may be administered in a single or divided dose, or for parenteral administration, it may be given as a continuous infusion.

[0077] Preferred embodiments of the present invention are presented below, wherein any combination of two or more of these embodiments is considered to be within the scope of the present invention.

[0078] That is, combinations of the preferred definitions of each substituent or moiety in formula (I) defined below are within the scope of the present invention.

[0079] According to a preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which L 1 is selected from a bond or an alkylene group.

[0080] In a preferred embodiment, the combination as defined above comprises a compound of formula (I) wherein L 1 is the key.

[0081] In a preferred embodiment, the combination as defined above comprises a compound of formula (I) wherein L 1 It is an alkylene group.

[0082] In another preferred embodiment, the combination as defined above comprises a compound of formula (I) wherein L 1 It's a carbonyl group.

[0083] In another preferred embodiment, the combination as defined above comprises a compound of formula (I) wherein L 1 Selected from -CH 2 -、-CHF-、-CF 2 -、-CH 2 -CH 2 .

[0084] In another more preferred embodiment, the combination as defined above comprises a compound of formula (I) wherein L 1 Yes-CH 2 -.

[0085] According to a preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 1 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkoxy, azido, hydroxyl, cyano, nitro, -NR 5 R 6 .

[0086] In particular, the combination according to the invention comprises compounds of formula (I) in which R 1 is selected from the group consisting of hydrogen atom, halogen atom, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkoxy, hydroxyl, -NR 5 R 6 .

[0087] However, another preferred embodiment provides a combination comprising a compound of formula (I) wherein R 1 and m>1. Each of the following is selected from the group consisting of hydrogen atom, halogen atom, optionally substituted alkyl group, haloalkyl group, fluoroalkyl group, optionally substituted alkoxy group and hydroxyl group, either alone or simultaneously or independently.

[0088] In another embodiment, the combination as defined above comprises a compound of formula (I) wherein R 1 m>1 alone or simultaneously or independently selected from a hydrogen atom, a halogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, and a hydroxyl group.

[0089] In another embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m> 1, simultaneously or independently selected from hydrogen atom, halogen atom, -NR 5 R 6 , where R 5 and R 6 may be selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, an optionally substituted heterocyclic group, or R 5 and R 6 The nitrogen atoms to which they are covalently attached may be linked together to form an optionally substituted heterocyclic group.

[0090] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m> 1, simultaneously or independently selected from hydrogen atom, halogen atom, -NR 5 R 6 , where R 5 and R 6 can be selected from a hydrogen atom, an optionally substituted alkyl group, or R 5 and R 6 The nitrogen atoms to which they are covalently attached may be linked together to form an optionally substituted heterocyclic group.

[0091] In another embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m> 1, simultaneously or independently selected from hydrogen atom, halogen atom, -NR 5 R 6 , where R 5 and R 6 It may be selected from a hydrogen atom, an optionally substituted alkyl group.

[0092] In particular, the combination according to the invention comprises compounds of formula (I) in which R 1 alone or when m> 1, simultaneously or independently selected from hydrogen atom, halogen atom, -NR 5 R6 , where R 5 and R 6 It can be selected from a hydrogen atom, a methyl group, an ethyl group, an isopropyl group, a tert-butyl group, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and a cyclohexyl group.

[0093] In particular, the combination according to the invention comprises compounds of formula (I) in which R 1 alone or when m> 1, simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 It can be selected from a hydrogen atom, a methyl group, and an optionally substituted heterocyclic group.

[0094] In particular, the combination according to the invention comprises compounds of formula (I) in which R 1 alone or when m> 1, simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 2 and R 3 The nitrogen atoms to which they are covalently attached may be linked together to form an optionally substituted heterocyclic group.

[0095] According to a preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 1 individually or when m>1 simultaneously or independently selected from a hydrogen atom or a halogen atom.

[0096] According to a more preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 1 It is selected from hydrogen atom, fluorine, chlorine, bromine, either alone or simultaneously or independently when m>1, more preferably hydrogen atom or chlorine.

[0097] However, according to a more preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 1 It is selected from hydrogen atom, fluorine, chlorine, bromine, either alone or simultaneously or independently when m>1, and most preferably is hydrogen atom or fluorine.

[0098] However, according to a more preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, fluorine.

[0099] However, according to a more preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, chlorine.

[0100] According to a preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 1 and m>1, alone or simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, a haloalkyl group, a fluoroalkyl group.

[0101] In a preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, alkyl.

[0102] In a more preferred embodiment, the combination according to the invention comprises a compound of formula (I), wherein R 1 individually or when m>1, simultaneously or independently selected from hydrogen atom, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl.

[0103] In a more preferred embodiment, the combination according to the invention comprises a compound of formula (I), wherein R 1 Individually or when m>1, simultaneously or independently selected from a hydrogen atom or a methyl group.

[0104] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, cycloalkyl.

[0105] However, in a more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 individually or when m>1 simultaneously or independently selected from hydrogen atom, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0106] However, another preferred embodiment provides a combination comprising a compound of formula (I) wherein R 1 individually or when m>1 simultaneously or independently selected from hydrogen atom, hydroxyl group, optionally substituted alkoxy group.

[0107] In a more preferred embodiment, the combination according to the invention comprises a compound of formula (I), wherein R 1 Individually or when m>1, simultaneously or independently selected from hydrogen atom, hydroxyl, methoxy, ethoxy, isopropoxy, sec-butoxy, tert-butoxy.

[0108] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 Individually or when m>1, simultaneously or independently selected from hydrogen atom, hydroxyl, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy.

[0109] However, in a more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, chlorine, hydroxyl, methoxy.

[0110] However, in another more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, chlorine, hydroxyl group.

[0111] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m> 1, simultaneously or independently selected from hydrogen atom, fluorine, chlorine, bromine, -CF 3 .

[0112] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 individually or when m>1, simultaneously or independently selected from hydrogen atom, fluorine, chlorine, bromine.

[0113] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, chlorine.

[0114] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 1 alone or when m>1, simultaneously or independently selected from hydrogen atom, cyano group.

[0115] According to a preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R 2 and R 3 and may be simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, and an optionally substituted heterocyclic group.

[0116] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, and an optionally substituted heterocyclic group.

[0117] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3They may be simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, and an optionally substituted heterocyclic group.

[0118] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom, an alkyl group, and a cycloalkyl group.

[0119] However, in another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom or a heterocyclic group.

[0120] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom and an alkyl group.

[0121] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom, a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, an isobutyl group, a sec-butyl group, and a tert-butyl group.

[0122] In a more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom and a tert-butyl group.

[0123] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and a cyclohexyl group.

[0124] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 They may be simultaneously or independently selected from a hydrogen atom and a heterocyclic group.

[0125] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 2 and R 3 The nitrogen atoms to which they are covalently attached may be linked together to form an optionally substituted heterocyclic group.

[0126] According to a preferred embodiment, the combination according to the invention comprises a compound of formula (I) in which R4 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkoxy, azido, hydroxyl, cyano, nitro, -NR 5 R 6 .

[0127] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 alone or when n>1, simultaneously or independently selected from hydrogen atom, halogen atom, -NR 5 R 6 , where R 5 and R 6 may be selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, an optionally substituted heterocyclic group, or R 5 and R 6 The nitrogen atoms to which they are covalently attached may be linked together to form an optionally substituted heterocyclic group.

[0128] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 It may be selected from a hydrogen atom, an optionally substituted alkyl group.

[0129] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 It may be selected from a hydrogen atom, an optionally substituted cycloalkyl group.

[0130] In a preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 can be selected from a hydrogen atom, a heterocyclic group, or R 5 and R 6 They may be linked together with the nitrogen atom to which they are covalently attached to form a heterocyclic group.

[0131] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R4 individually or when n>1 simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 They may be linked together with the nitrogen atom to which they are covalently attached to form a heterocyclic group.

[0132] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 It may be selected from a hydrogen atom or form a heterocyclic group.

[0133] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 It can be selected from a hydrogen atom, a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, an isobutyl group, a sec-butyl group, and a tert-butyl group.

[0134] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom, -NR 5 R 6 , where R 5 and R 6 It can be selected from a hydrogen atom, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and a cyclohexyl group.

[0135] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from a hydrogen atom, a halogen atom, an optionally substituted alkyl group.

[0136] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 Individually or when n>1, simultaneously or independently, they are selected from a hydrogen atom or a cycloalkyl group.

[0137] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 alone or when n>1, simultaneously or independently selected from hydrogen atom, alkyl.

[0138] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl.

[0139] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 Individually or when n>1, simultaneously or independently selected from hydrogen atom, halogen atom, alkyl group, halogenated alkyl group, fluoroalkyl group, alkoxy group, hydroxyl group.

[0140] In another more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 alone or when n>1, simultaneously or independently selected from hydrogen atom, alkoxy group.

[0141] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 Individually or when n>1, simultaneously or independently selected from hydrogen atom, hydroxyl, methoxy, ethoxy, isopropoxy, sec-butoxy, tert-butoxy.

[0142] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 Individually or when m>1, simultaneously or independently selected from hydrogen atom, hydroxyl, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy.

[0143] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 alone or when m>1, simultaneously or independently selected from hydrogen atom, methoxy group.

[0144] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 Individually or when n>1, simultaneously or independently selected from hydrogen atom, hydroxyl group, methoxy group.

[0145] In another embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 Individually or when n>1, simultaneously or independently selected from hydrogen atom, hydroxyl group, methyl group, methoxy group.

[0146] In another more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 alone or when n>1, simultaneously or independently selected from hydrogen atom, fluorine, chlorine, bromine, -CF 3 .

[0147] In another preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from hydrogen atom or halogen.

[0148] In another more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 alone or when n>1, simultaneously or independently selected from hydrogen atom, fluorine, chlorine.

[0149] In another more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1 simultaneously or independently selected from a hydrogen atom or a cyano group.

[0150] In another more preferred embodiment, the combination according to the invention comprises a compound of formula (I) wherein R 4 individually or when n>1, simultaneously or independently, are hydrogen atoms.

[0151] In another embodiment, the combination according to the invention comprises

[0152] ♦ A compound selected from the group consisting of compounds of formula (II), (III) and (IV) as shown below:

[0153]

[0154]

[0155]

[0156] in

[0157] • R 1 is selected from the group consisting of hydrogen atom, halogen atom, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted alkoxy, -OR 7 , azido, hydroxy, cyano, nitro, -NR 5 R 6 ;

[0158] • R 2 and R 3 are simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted heterocyclic group, or R 2 and R 3may be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group;

[0159] • R 4 is selected from the group consisting of hydrogen atom, halogen atom, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted alkoxy, -OR 7 , azido, hydroxy, cyano, nitro, -NR 5 R 6 .

[0160] • R 5 and R 6 are simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted heterocyclic group, or R 5 and R 6 may be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group;

[0161] • R 7 may be selected from optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocyclyl;

[0162] • n is an integer which can have any value of 0, 1, 2, 3 or 4;

[0163] • m is an integer which for formula (II) may have any value of 0, 1, 2, 3, 4 or 5; and

[0164] • m is an integer which may have any value of 0, 1, 2, 3 or 4 for formulae (III) and (IV);

[0165] and pharmaceutically acceptable salts, hydrates, solvates, prodrugs, polymorphs, tautomers, isotopic variants, isomers, stereoisomers or mixtures of stereoisomers, and

[0166] ♦Mitogen-activated protein kinase kinase inhibitors (MEK inhibitors) and pharmaceutically acceptable salts, hydrates or solvates thereof.

[0167] The compounds of formula (II), (III) and (IV) are preferred embodiments of the compounds of formula (I).

[0168] All definitions of terms and alternative preferred embodiments described above in connection with formula (I) also apply to formula (II), (III) and (IV).

[0169] In particular, the combination according to the invention comprises a compound of formula (I), wherein

[0170] ·L 1 Selected from single bonds and (-CH 2 -) p Group;

[0171] ·R 1 are halogen atoms, especially chlorine;

[0172] ·R 2 and R 3 are simultaneously or independently selected from a hydrogen atom and an optionally substituted alkyl group, in particular a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group.

[0173] m = 1,

[0174] n = 0, and

[0175] p = 1,

[0176] and pharmaceutically acceptable salts, hydrates, solvates, prodrugs, polymorphs, tautomers, isotopic variations, isomers, stereoisomers or mixtures of stereoisomers thereof.

[0177] In a preferred embodiment, the combination according to the invention comprises a compound selected from the group consisting of:

[0178] 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (1-5) of formula (Ia)

[0179]

[0180] or a pharmaceutically acceptable salt, solvate or prodrug thereof, and

[0181] 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib)

[0182]

[0183] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0184] In particular, the combination according to the invention comprises a compound selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (1-6) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (2-3).

[0185] The preparation of compounds of formula (I), (II), (III) and (IV) is described in WO2016 / 067112 and WO2020 / 048694. In particular, compounds 1-5, 1-6, 2-2 and 2-3 are described in Examples 1 and 2 of WO2016 / 067112 and WO2020 / 048694.

[0186] In a specific embodiment of the present invention, the MEK inhibitor can be selected from Avutometinib (RO-5126766) [946128-88-7], BI-847325 [1207293-36-4], bemetinib [606143-89-9], cobimetinib [934660-93-2], selumetinib [606143-52-6], trametinib [871700-17-3], AZD8330 [869357-68-6], BI-847325 [1207293-36-4], GDC-0623 [1168091-68-6]、Mirdametinib(PD0325901)[391210-10-9]、PD184352(CI-1040)[212631-79-3]、Pimasertib(AS-703026)[1236699-92-5]、Rifatinib[923032-37-5]、TAK-733 [1035555-63-5]、Tunlametinib(HL-085)[1801756-06-8]、BIX 02188 [1094614-84-2]、BIX 02189 [1265916-41-3], E6201 [603987-35-5], FCN-159, GDC-0623 [1168091-68-6], magnolol [35354-74-6], myricetin [529-44-2], PD98059 [167869-21-8], PD184352 (CI-1040) [212631-79-3], PD318088 [391210-00-7], SL-327 [305350-87-2], SHR7390, TAK-733 [1035555-63-5], U0126 [109511-58-2].

[0187] In other embodiments of the present invention, the MEK inhibitor can be selected from bemetinib [606143-89-9], cobimetinib [934660-93-2], selumetinib [606143-52-6], trametinib [871700-17-3], AZD8330 [869357-68-6], BI-847325 [1207293-36-4], GDC-0623 [1168091-68-6], Mirdametinib (PD0325901) [391210-10-9], PD184352 (CI-1040) [212631-79-3], Pimasertib (AS-703026) [1236699-92-5], Rifatinib [923032-37-5], TAK-733 [1035555-63-5], BIX 02188 [1094614-84-2], BIX 02189 [1265916-41-3], Honokiol [35354-74-6], Myricetin [529-44-2], PD98059 [167869-21-8], PD318088 [391210-00-7], SL-327 [305350-87-2], U0126 [109511-58-2] and pharmaceutically acceptable salts, hydrates or solvates thereof.

[0188] In a preferred embodiment, the MEK inhibitor can be selected from bemetinib [606143-89-9], cobimetinib [934660-93-2], selumetinib [606143-52-6], trametinib [871700-17-3], AZD8330 [869357-68-6], BI-847325 [1207293-36-4], GDC-0623 [1168091-68-6], Mirdametinib (PD0325901) [391210-10-9], PD184352 (CI-1040) [212631-79-3], Pimasertib (AS-703026) [1236699-92-5], Rifatinib [923032-37-5], TAK-733 [1035555-63-5].

[0189] In another preferred embodiment, the MEK inhibitor can be selected from bemetinib [606143-89-9], cobimetinib [934660-93-2], selumetinib [606143-52-6], and trametinib [871700-17-3].

[0190] In another preferred embodiment, the MEK inhibitor can be selected from bemetinib [606143-89-9], cobimetinib [934660-93-2], selumetinib [606143-52-6], trametinib [871700-17-3] and pharmaceutically acceptable salts, hydrates or solvates thereof.

[0191] In another preferred embodiment, the MEK inhibitor is trametinib [871700-17-3] or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0192] In another preferred embodiment, the MEK inhibitor is cobimetinib [934660-93-2] or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0193] In another embodiment, the MEK inhibitor is trametinib [871700-17-3].

[0194] In a preferred embodiment, the MEK inhibitor is the dimethyl sulfoxide solvate form of trametinib [1187431-43-1].

[0195] In another embodiment, the MEK inhibitor is cobimetinib [934660-93-2].

[0196] In a preferred embodiment, the MEK inhibitor is cobimetinib hemifumarate [1369665-02-0].

[0197] In particular, the combination according to the present invention comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a salt, solvate or prodrug thereof and trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof.

[0198] In another embodiment, the combination according to the invention comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a salt, solvate or prodrug thereof and trametinib in the form of a dimethyl sulfoxide solvate [1187431-43-1].

[0199] In another embodiment, the combination according to the present invention comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a salt, solvate or prodrug thereof and cobimetinib [934660-93-2] or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0200] In another embodiment, the combination according to the present invention comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a salt, solvate or prodrug thereof and cobimetinib hemifumarate [1369665-02-0] or a hydrate or solvate thereof.

[0201] According to one embodiment, the present invention relates to a combination consisting of the following components as described above with respect to all general or specific embodiments:

[0202] ♦ a compound of formula (I), (II), (III) or (IV) and a pharmaceutically acceptable salt, hydrate, solvate, prodrug, polymorph, tautomer, isotopic variant, isomer, stereoisomer or mixture of stereoisomers thereof, and

[0203] ♦Mitogen-activated protein kinase kinase inhibitors (MEK inhibitors) and pharmaceutically acceptable salts or solvates thereof.

[0204] The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a combination as defined above, said combination comprising a compound of formula (I), (II), (III) or (IV) and a pharmaceutically acceptable salt, hydrate, solvate, prodrug, polymorph, tautomer, isotopic variant, isomer, stereoisomer or mixture of stereoisomers and a MEK inhibitor and a pharmaceutically acceptable salt and solvate thereof, and at least one pharmaceutically acceptable carrier, wherein each component of the combination may be contained in a separate pharmaceutical composition.

[0205] The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a combination as defined above, said combination comprising a compound of formula (I), (II), (III) or (IV) and a pharmaceutically acceptable salt, hydrate, solvate, prodrug, polymorph, tautomer, isotopic variant, isomer, stereoisomer or mixture of stereoisomers and a MEK inhibitor and a pharmaceutically acceptable salt and solvate thereof, and at least one pharmaceutically acceptable carrier, wherein each component of the combination may be contained in the same pharmaceutical composition.

[0206] That is, the components of the combination, which are the active ingredients as defined above (the compound of formula (I), (II), (III) or (IV) and the MEK inhibitor), can be administered simultaneously (i.e. concurrently) in the same or different pharmaceutical compositions, or independently in different pharmaceutical compositions, or sequentially in different pharmaceutical compositions in any order.

[0207] Each pharmaceutical composition containing the components of the combination may be administered once or several times in one or different doses, using the same or different administration routes.

[0208] The amounts, relative timings and routes of administration of the active ingredients will be selected so as to achieve the desired combined therapeutic effect.

[0209] According to a preferred embodiment, each component of the combination may be contained in a separate pharmaceutical composition.

[0210] According to another preferred embodiment, each component of the combination may be contained in the same pharmaceutical composition.

[0211] All general and preferred embodiments described above for the combination of a compound of formula (I), (II), (III) or (IV) and a MEK inhibitor and for each component of this combination also apply to the pharmaceutical composition.

[0212] The pharmaceutical compositions of the invention may be suitable for oral, parenteral, ocular, transdermal or nasal administration, or for inhalation.

[0213] The present invention also relates to a combination comprising a compound of formula (I) as defined above or a pharmaceutically acceptable salt or solvate thereof and a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0214] The present invention also relates to a combination comprising a compound of formula (II), (III) or (IV) as defined above or a pharmaceutically acceptable salt or solvate thereof and a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0215] The present invention also relates to a combination comprising a compound of formula (II), (III) or (IV) as defined above or a pharmaceutically acceptable salt or solvate thereof and a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular intrahepatic bile duct carcinoma and / or extrahepatic bile duct carcinoma.

[0216] The present invention also relates to a combination comprising a compound of formula (II), (III) or (IV) as defined above or a pharmaceutically acceptable salt or solvate thereof and a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular intrahepatic bile duct carcinoma.

[0217] The present invention also relates to a combination comprising a compound of formula (I) selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (1-5) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0218] The present invention also relates to a combination comprising a compound of formula (I) selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (1-6) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (2-3) and a MEK inhibitor as defined above, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0219] The present invention also relates to a combination comprising a compound of formula (I) selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (1-5) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0220] The present invention also relates to a combination comprising a compound of formula (I) selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (1-6) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (2-3) and a MEK inhibitor as defined above, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0221] In another aspect, the present invention relates to a combination comprising a compound of formula (I) selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (1-5) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer.

[0222] In yet another aspect, the present invention relates to a combination comprising a compound of formula (I) selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (1-6) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (2-3) and a MEK inhibitor as defined above, for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer.

[0223] In another aspect, the present invention also relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0224] The present invention also relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0225] In yet another aspect, the present invention also relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above for use in treating and / or preventing and / or reducing liver cancer-related diseases, particularly intrahepatic bile duct carcinoma.

[0226] In yet another aspect, the present invention also relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular extrahepatic bile duct cancer.

[0227] In another aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) and a MEK inhibitor as defined above for use in treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer.

[0228] In another aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof for treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0229] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof for treating and / or preventing and / or reducing liver cancer.

[0230] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof for treating and / or preventing and / or reducing bile duct cancer, wherein the bile duct cancer is intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0231] In another aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and a 1:1 dimethyl sulfoxide (DMSO) solvate form of trametinib [1187431-43-1] for treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0232] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and a 1:1 dimethyl sulfoxide (DMSO) solvate form of trametinib [1187431-43-1] for use in treating and / or preventing and / or reducing liver cancer.

[0233] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and a 1:1 dimethyl sulfoxide (DMSO) solvate form of trametinib [1187431-43-1] for treating and / or preventing and / or reducing bile duct cancer, wherein the bile duct cancer is intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0234] More preferably, the combination comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof.

[0235] In yet another aspect, the combination comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt, solvate or prodrug thereof and a 1:1 dimethyl sulfoxide (DMSO) solvate form of trametinib [1187431-43-1].

[0236] In another aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib [934660-93-2] or a pharmaceutically acceptable salt or solvate thereof for treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0237] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt, solvate or prodrug thereof and cobimetinib [934660-93-2] or a pharmaceutically acceptable salt or solvate thereof for treating and / or preventing and / or reducing liver cancer.

[0238] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib [934660-93-2] or a pharmaceutically acceptable salt or solvate thereof for treating and / or preventing and / or reducing bile duct cancer, wherein the bile duct cancer is intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0239] In another aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib hemifumarate [1369665-02-0] for treating and / or preventing and / or reducing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0240] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib hemifumarate [1369665-02-0] for treating and / or preventing and / or reducing liver cancer.

[0241] In another preferred aspect, the present invention relates to a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib hemifumarate [1369665-02-0] for treating and / or preventing and / or reducing bile duct cancer, wherein the bile duct cancer is intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0242] More preferably, the combination comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib [934660-93-2] or a pharmaceutically acceptable salt or solvate thereof.

[0243] In yet another aspect, the combination comprises 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib hemifumarate [1369665-02-0].

[0244] In a preferred embodiment, the liver cancer-related disease is cholangiocarcinoma.

[0245] In a more preferred embodiment, the liver cancer-related disease is selected from intrahepatic bile duct carcinoma and / or extrahepatic bile duct carcinoma.

[0246] In another preferred embodiment, the liver cancer-related disease is intrahepatic cholangiocarcinoma.

[0247] In another preferred embodiment, the liver cancer-related disease is extrahepatic bile duct cancer.

[0248] In another preferred embodiment, the liver cancer-related disease is liver cancer.

[0249] In another preferred embodiment, the liver cancer-related disease is hepatoblastoma.

[0250] According to the present invention, each component of the combination may be used simultaneously, independently or sequentially in the therapy of liver cancer.

[0251] According to the present invention, the compounds of formula (I), (II), (III) or (IV) as defined above and the MEK inhibitors as defined above and pharmaceutical compositions containing them (wherein each component of the composition may be contained in an independent pharmaceutical composition) can be administered to a human or animal in need thereof as a combination therapy in the treatment and / or reduction and / or prevention of liver cancer, wherein the administration may be simultaneous, independent or sequential.

[0252] All general and preferred embodiments described above for the combination of a compound of formula (I), (II), (III) or (IV) and a MEK inhibitor and for each component of such a combination also apply to their use in the treatment and / or prevention and / or reduction of liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0253] In another aspect, the present invention also relates to a method for treating and / or reducing and / or preventing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma, the method comprising administering to a patient in need thereof a combination of a compound of formula (I), (II), (III) or (IV) or a pharmaceutically acceptable salt or solvate thereof and a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof as described above.

[0254] In another aspect, the present invention also relates to a method for treating and / or reducing and / or preventing liver cancer-related diseases, in particular bile duct cancer, wherein the bile duct cancer is intrahepatic bile duct cancer and / or extrahepatic bile duct cancer.

[0255] All general and preferred embodiments described above for the combination of a compound of formula (I), (II), (III) or (IV) and a MEK inhibitor and each component of such a combination also apply to the method for treating and / or reducing and / or preventing liver cancer-related diseases, in particular liver cancer, hepatoblastoma or cholangiocarcinoma.

[0256] The present invention also relates to a kit comprising a combination of a compound of formula (I), (II), (III) or (IV) as defined above, or a pharmaceutically acceptable salt or solvate thereof, and a MEK inhibitor, or a pharmaceutically acceptable salt or solvate thereof, wherein the two or either component is in the form of a pharmaceutical composition that can be administered simultaneously, independently or sequentially.

[0257] The invention is illustrated, but not limited, by the following examples.

[0258] Example

[0259] ■ Example 1: In vitro evaluation of the cytotoxic activity of compound 2-2, hydroxychloroquine (HCQ), trametinib and cobimetinib in the intrahepatic cholangiocarcinoma cell line HuCCT1 (KRAS G12D status)

[0260] Cell culture: HuCCT1 cell line carrying KRAS G12D mutation (JCRB Cell Bank #JCRB0425) was maintained in RPMI medium (Dutscher #L0498-500) containing 1% penicillin-streptomycin (Dutscher #P06-07100) and 10% fetal bovine serum (Dutscher #SV30160.03C) in a humidified cell culture incubator at 37°C and 5% CO 2 Next cultivation.

[0261] HuCCT1 cells were plated at 4.10 μl per well. 3 Cells were plated at a density of 10 cells / mL in Greiner Bio-one µClear® P96-well plates (Dutscher #655098) containing 80 µL of medium. After 24 hours, HuCCT1 cells were treated with compound 2-2, hydroxychloroquine sulfate (HCQ, [747-36-4]), trametinib (Selleckchem #S2673, [871700-17-3]), or cobimetinib (Selleckchem #S8041, [934660-93-2]). Cell viability was assessed 72 hours after molecular treatment by the CellTiter-Glo® Luminescent Cell Viability Assay (Promega #G7573) according to the manufacturer's protocol and using an Infinity F200 Pro Luminometer (Tecan). DMSO was used as a negative control for compound 2-2, trametinib, and cobimetinib, and water was used as a negative control for HCQ. IC 50 The values ​​were determined as the compound dose required to reduce the luminescence value to 50% of the reference signal obtained for untreated cell cultures. Raw data were analyzed using GraphPad Prism software v9.4 (GraphPad Software, Inc. La Jolla, CA). For all experiments, the analytical data are shown as the average of three parallel samples and four independent experiments. The four compounds 2-2, HCQ, trametinib and cobimetinib showed dose-responsive cytotoxic activity against the intrahepatic cholangiocarcinoma cell line HuCCT1 (Table 1).

[0262] surface : Cell viability assay of compound 2-2, hydroxychloroquine (HCQ), trametinib and cobimetinib on intrahepatic cholangiocarcinoma cell line HuCCT1 (IC 50 , µM) Data represent the mean ± SD of at least three independent experiments performed in triplicate.

[0263] Example 2: In vitro evaluation of the combined effect of the binary drug-drug combination of compound 2-2 + trametinib and hydroxychloroquine (HCQ) + trametinib on cytotoxic activity in the intrahepatic cholangiocarcinoma cell line HuCCT1 (KRAS G12D status)

[0264] HuCCT1 cell viability assay using binary drug-drug combinations was performed as described in Example 1. Binary drug-drug combinations of compound 2-2 + trametinib and HCQ + trametinib were tested in a checkerboard format in Greiner Bio-one µClear® 96-well plates (Dutscher #655098). The concentration range was chosen to be within the IC of each individual compound. 50 Values ​​near and around. Compound 2-2 in combination with trametinib was tested at 1.28 µM, 1.6 µM, 2.0 µM, 2.5 µM, and 3.125 µM. HCQ [747-36-4] in combination with trametinib was tested at 15 µM, 20 µM, 30 µM, 45 µM, 60 µM. Trametinib (Selleckchem #S2673, [871700-17-3]) in combination with compound 2-2 or in combination with HCQ was tested at 0.005 µM, 0.03 µM, 6.0 µM, 15 µM, 30 µM, 60 µM, 90 µM in both cases. Two softwares were used: MacSynergy TM The combined effects of two binary drug-drug combinations, 2-2 + trametinib and HCQ + trametinib, were analyzed using MacSynergy II (1) and SynergyFinder (2). TM II was used to analyze the combined effects according to the Bliss independence model, which assumes probabilistic statistical independence between the combined compounds (3). The main assumption of the Bliss independence principle is that two or more combined drugs act independently of each other in different modes of action. SynergyFinder was used to analyze the combined effects according to the Loewe additivity model, which is based on its principle of pseudodrug-drug combination, which assumes that there is no interaction when a compound is combined with itself (4).

[0265] Joint effect analysis based on Loewe's additive model (SynergyFinder):

[0266] SynergyFinder was used to evaluate the combined effects of the drug-drug combinations of compound 2-2 + trametinib (Table 2) and HCQ + trametinib (Table 3) for their individual abilities to inhibit the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Loewe additivity model.

[0267] Table 2: Analysis of the combined effect of the drug-drug combination of compound 2-2 + trametinib on the intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Loewe additive model (SynergyFinder) # # Synergy scores (X) obtained from SynergyFinder were defined as follows: values ​​of X < -10 reflect antagonism; values ​​of -10 < X ​​< 10 indicate additivity; and values ​​of X > 10 indicate synergy (2). Data represent the mean ± SD of four independent experiments performed in triplicate.

[0268] When the drug-drug combination of compound 2-2 + trametinib was analyzed using SynergyFinder for its ability to inhibit the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 by inducing a cytotoxic effect (assessed by the CellTiter-Glo® luminescent cell viability assay), the drug combination showed an overall additive combined effect (-10 < synergy score < +10) according to the Loewe additive model (Table 2). According to the Loewe dependent additive model, the overall additive combined effect of the drug-drug combination of compound 2-2 + trametinib did not substantially diverge from the different modes of action of the two compounds (i.e., compound 2-2 as an autophagy inhibitor and trametinib as a MEK inhibitor).

[0269] Table 3: Analysis of the combined effects of drug-drug combination of hydroxychloroquine + trametinib on intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Loewe additive model (SynergyFinder) # # Synergy scores (X) obtained from SynergyFinder were defined as follows: values ​​of X < -10 reflect antagonism; values ​​of -10 < X ​​< 10 indicate additivity; and values ​​of X > 10 indicate synergy (2). Data represent the mean ± SD of four independent experiments performed in triplicate.

[0270] When the drug-drug combination of hydroxychloroquine + trametinib was analyzed using SynergyFinder for its ability to inhibit the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 by inducing a cytotoxic effect (assessed by the CellTiter-Glo® luminescent cell viability assay), the drug combination showed an overall additive combined effect (-10 < synergy score < +10) according to the Loewe dependent additivity model (Table 3). The overall additive combined effect of the drug-drug combination of hydroxychloroquine + trametinib did not substantially diverge from the different modes of action of the two compounds (i.e., hydroxychloroquine as an autophagy inhibitor and trametinib as a MEK inhibitor) according to the Loewe dependent additivity model.

[0271] Combined effect analysis based on the Bliss independence model (MacSynergy TM II):

[0272] According to the Bliss independence model, using MacSynergy TM II The combined effects of the drug-drug combinations of compound 2-2 + trametinib and hydroxychloroquine + trametinib were evaluated for their respective abilities to inhibit the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 (Table 4).

[0273] Table 4: Analysis of the combined effects of the drug-drug combination of compound 2-2 + trametinib and hydroxychloroquine + trametinib on the intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Bliss independence model (MacSynergy TM II) # Synergy plot at 95% confidence interval. To explain the TM II generated synergy values ​​for the amount of synergy and the degree of antagonism, and developed general guidelines as follows: TM II software in the positive or negative direction 0 to 25, 25 to 50, 50 to 100 and >100 μm 2 The % calculated values ​​were defined as insignificant synergy or antagonism, slight synergy or antagonism, moderate synergy or antagonism, or strong synergy or antagonism, respectively (5). Data represent the mean ± SD of four independent experiments performed in triplicate.

[0274] According to the Bliss independence model, the binary drug-drug combination of compound 2-2 + trametinib showed significant moderate synergy in inhibiting the growth of intrahepatic cholangiocarcinoma cell line HuCCT1 (synergy score = 62.8 ± 23.9 µM².%, Table 4), with potential significant effects in vivo and clinical settings. The combination of compound 2-2 + trametinib did not show antagonistic regions (antagonism score = -7.2 ± 7.20 µM².%, Table 4). In contrast, according to the Bliss independence model, the binary drug-drug combination of hydroxychloroquine + trametinib showed a non-significant synergistic drug-drug combination effect (synergy score = 18.3 ± 10.6 µM².%, Table 4), and had an overall additive effect on inhibiting the growth of intrahepatic cholangiocarcinoma cell line HuCCT1 (5, 6). Furthermore, the combination of hydroxychloroquine + trametinib showed insignificant antagonism comparable to its synergy score (synergy score = -18.3 ± 10.6 µM².%, antagonism score = -21.5 ± 2.9 µM².%, see Table 4).

[0275] Both compound 2-2 and hydroxychloroquine are known autophagy inhibitors in the art (7, 8). However, when these two autophagy inhibitors were combined with the MEK inhibitor trametinib to inhibit the growth of intrahepatic cholangiocarcinoma HuCCT1 cell line, hydroxychloroquine only showed an additive effect, while compound 2-2 showed a significant synergistic combination effect.

[0276] Example 3: In vitro evaluation of the cytotoxic activity of the binary drug-drug combination of compound 2-2 + cobimetinib and hydroxychloroquine (HCQ) + cobimetinib in the intrahepatic cholangiocarcinoma cell line HuCCT1 (KRAS G12D status)

[0277] HuCCT1 cell viability assay using binary drug-drug combinations was performed as described in Example 1. Binary drug-drug combinations of compound 2-2 + cobimetinib and HCQ + cobimetinib were tested in a checkerboard format in Greiner Bio-one µClear® 96-well plates (Dutscher #655098). The concentration range was chosen to be within the IC of each individual compound. 50values. Compound 2-2 in combination with cobimetinib was tested at 1.28 µM, 1.6 µM, 2.0 µM, 2.5 µM, and 3.125 µM. HCQ [747-36-4] in combination with cobimetinib was tested at 15 µM, 20 µM, 30 µM, 45 µM, 60 µM. Cobimetinib (Selleckchem #S8041, [934660-93-2]) in combination with compound 2-2 or in combination with HCQ was tested at 0.10 µM, 1.0 µM, 10 µM, 30 µM, 60 µM, 80 µM, 100 µM in both cases. TM The combined effects of two binary drug-drug combinations, 2-2 + cobimetinib and HCQ + cobimetinib, were analyzed using MacSynergy II (1) and SynergyFinder (2). TM II was used to analyze the combined effects according to the Bliss independence model, which assumes probabilistic statistical independence between the combined compounds (3). SynergyFinder was used to analyze the combined effects according to the Loewe additivity model, which is based on its principle of pseudodrug-drug combination, assuming that there is no interaction when a compound is combined with itself (4).

[0278] Joint effect analysis based on Loewe's additive model (SynergyFinder):

[0279] SynergyFinder was used to evaluate the combined effects of binary drug-drug combinations of compound 2-2 + cobimetinib (Table 5) and HCQ + cobimetinib (Table 6) for their respective abilities to inhibit the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Loewe additivity model.

[0280] Table 5: Analysis of the combined effect of the drug-drug combination of compound 2-2 + cobimetinib on the intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Loewe additive model (SynergyFinder) # # Synergy scores (X) obtained from SynergyFinder were defined as follows: values ​​of X < -10 reflect antagonism; values ​​of -10 < X ​​< 10 indicate additivity; and values ​​of X > 10 indicate synergy (2). Data represent the mean ± SD of three independent experiments performed in triplicate.

[0281] When the drug-drug combination of compound 2-2 + cobimetinib was analyzed using SynergyFinder for its combined properties in inhibiting the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 by inducing a cytotoxic effect (which was assessed by the CellTiter-Glo® luminescent cell viability assay), the drug combination showed an overall additive combined effect (-10 < synergy score < +10) according to the Loewe additivity model (Table 5). Advantageously, when the binary drug-drug combination of compound 2-2 + cobimetinib was analyzed according to the Bliss additivity model, the binary combination showed a significant synergistic combined effect (50 µM².% < synergy score < 100 µM².%, see Table 7).

[0282] When analyzed using SynergyFinder according to the Loewe dependent additivity model, the overall additive combined effect of the drug-drug combination of compound 2-2 + cobimetinib (-10 < synergy score < +10) did not substantially differ from the distinct modes of action of the two compounds (i.e., compound 2-2 as a known autophagy inhibitor and cobimetinib as a MEK inhibitor).

[0283] Table 6: Analysis of the combined effects of drug-drug combination of hydroxychloroquine + cobimetinib on intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Loewe additive model (SynergyFinder) # # Synergy scores (X) obtained from SynergyFinder were defined as follows: values ​​of X < -10 reflect antagonism; values ​​of -10 < X ​​< 10 indicate additivity; and values ​​of X > 10 indicate synergy (2). Data represent the mean ± SD of three independent experiments performed in triplicate.

[0284] When the binary drug-drug combination of hydroxychloroquine + cobimetinib was analyzed using SynergyFinder for its ability to inhibit the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 by inducing a cytotoxic effect (assessed by the CellTiter-Glo® luminescent cell viability assay), the drug combination showed an overall additive combined effect (-10 < synergy score < +10) according to the Loewe dependent additivity model (Table 6). According to the Loewe dependent additivity model, the overall additive combined effect of the binary drug-drug combination of hydroxychloroquine + trametinib did not substantially diverge from the different modes of action of the two compounds, i.e., hydroxychloroquine as an autophagy inhibitor and cobimetinib as a MEK inhibitor.

[0285] Combined effect analysis based on the Bliss independence model (MacSynergy TMII):

[0286] According to the Bliss independence model, using MacSynergy TM II The combined effects of the binary drug-drug combinations of compound 2-2 + cobimetinib and hydroxychloroquine + cobimetinib were evaluated for their respective abilities to inhibit the growth of the intrahepatic cholangiocarcinoma cell line HuCCT1 (Table 7).

[0287] Table 7: Analysis of the combined effects of the drug-drug combination of compound 2-2 + cobimetinib and hydroxychloroquine + cobimetinib on the intrahepatic cholangiocarcinoma cell line HuCCT1 according to the Bliss independence model (MacSynergy TM II) # Synergy plot at 95% confidence interval. To explain the TM II generated synergy values ​​for the amount of synergy and the degree of antagonism, and developed general guidelines as follows: TM II software in the positive or negative direction 0 to 25, 25 to 50, 50 to 100 and >100 μm 2 The % calculated values ​​were defined as insignificant synergy or antagonism, slight synergy or antagonism, moderate synergy or antagonism, or strong synergy or antagonism, respectively (5). Data represent the mean ± SD of four independent experiments performed in triplicate.

[0288] According to the Bliss independence model, the binary drug-drug combination of compound 2-2 + cobimetinib showed overall significant moderate synergy in inhibiting the growth of intrahepatic cholangiocarcinoma cell line HuCCT1, with potential significant effects in vivo and clinical settings (synergy score = 90.2 ± 8.40 µM².%, Table 7). In addition, the binary drug-drug combination of compound 2-2 + cobimetinib showed no antagonistic space (antagonism score = -1.00 ± 1.62 µM².%, Table 7). In contrast, according to the Bliss independence model, the binary drug-drug combination of hydroxychloroquine + cobimetinib showed a slight synergistic drug-drug combination effect on inhibiting the growth of intrahepatic cholangiocarcinoma cell line HuCCT1 (synergy score = 39.8 ± 2.77 µM².%, Table 7), which may have a slight effect or even no effect in vivo (5,6). The binary drug-drug combination of hydroxychloroquine + cobimetinib showed no antagonism in the concentration space tested (antagonism score = -2.03 ± 0.78 µM².%, Table 7).

[0289] Both compound 2-2 and hydroxychloroquine are known autophagy inhibitors in the art (7, 8). However, when these two autophagy inhibitors were combined with the MEK inhibitor cobimetinib to inhibit the growth of intrahepatic cholangiocarcinoma HuCCT1 cell line, hydroxychloroquine only showed an overall additive effect according to the Loewe model, a slight synergistic effect according to the Bliss model, and no significant effect in vivo, while compound 2-2 showed an overall additive effect according to the Loewe additive model and a significant synergistic combined effect according to the Bliss independence model.

[0290] The present invention discloses that compound 2-2, when combined with a MEK inhibitor (e.g., trametinib, cobimetinib), exhibits a significant synergistic combined effect according to the Bliss independence model to inhibit the in vitro growth of cholangiocarcinoma, as demonstrated in the cell line HuCCT1 (intrahepatic cholangiocarcinoma cell line, Cellosaurus ID: CVCL_0324).

[0291] References

[0292] 1. Prichard, MN and Shipman, C. (1990). A three-dimensional model to analyze drug-drug interactions. Antiviral Res. 14, 181–205.

[0293] 2. Zheng, S., Wang, W., Aldahdooh, J., Malyutina, A., Shadbahr, T., Tanoli, Z., Pessia, A., and Tang, J. (2022). SynergyFinder Plus: Toward Better Interpretation and Annotation of Drug Combination Screening Datasets. Genomics. ProteomicsBioinformatics. 10.1016 / J.GPB.2022.01.004.

[0294] 3. Bliss, CI, (1939), THE TOXICITY OF POISONSAPPLIED JOINTLY1, Ann. Appl. Biol. 26, 585–615

[0295] 4. Loewe, S. (1953). The problem of synergism and antagonism of combined drugs. Arzneimittelforschung. 3, 285–290.

[0296] 5. Smee, DF and Prichard, MN (2017). Comparison of three dimensional synergistic analyses of percentage versus logarithmic data in antiviral studies. Antiviral Res. 145, 1–5.

[0297] 6. Prichard, MN and Shipman Jr., C. (1990). A three-dimensional model to analyze drug-drug interactions. Antivir. Res. 14, 181–205.

[0298] 7. Halfon, P., Bassissi, F., Brun, S., Courcambeck, J. and Rachid, M., (2020), Substituted 2,4 diamino-quinoline as new medicamentfor fibrosis, autophagy and cathepsins B (CTSB), L (CTSL) AND D (CTSD)related diseases, WO2020048694

[0299] 8. Pasquier, B. (2016). Autophagy inhibitors. Cell. Mol. Life Sci. 73, 985–1001.

Claims

1. A combination comprising: ♦Compounds of formula (I) in • L1 is selected from a single bond, optionally substituted (CH2) p group, optionally substituted alkylene, carbonyl; • R1 is selected alone or simultaneously or independently when m>1 from a hydrogen atom, a halogen atom, an optionally substituted alkyl group, a haloalkyl group, a fluoroalkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted alkoxy group, -O-R7, -O-(CO)-R7, -O-(CO)-NR5R6, -NR5-(CO)-R7, -O-(CO)-O-R7, -NR5-(CO)-O-R7, an azido group, a hydroxyl group, a cyano group, a nitro group, -NR5R6; • R2 and R3 are simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted heterocyclic group, an optionally substituted aryl group, an optionally substituted benzyl group, an optionally substituted heteroaryl group; Alternatively, R2 and R3 may be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group; • R4 is selected, alone or when n>1, simultaneously or independently, from a hydrogen atom, a halogen atom, an optionally substituted alkyl group, a haloalkyl group, a fluoroalkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted alkoxy group, -O-R7, -O-(CO)-R7, -O-(CO)-NR5R6, -NR5-(CO)-R7, -O-(CO)-O-R7, -NR5-(CO)-O-R7, an azido group, a hydroxyl group, a cyano group, a nitro group, -NR5R6; • R5 and R6 are simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted heterocyclic group, an optionally substituted aryl group, an optionally substituted heteroaryl group; Alternatively, R5 and R6 may be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group; • R7 may be selected from optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl; • n is an integer which can have any value of 0, 1, 2, 3 or 4; • m is an integer which can have any value of 0, 1, 2, 3, 4 or 5; • p is an integer which can have any value of 0 or 1; and pharmaceutically acceptable salts, hydrates, solvates, polymorphs, tautomers, isotopic variants, stereoisomers or mixtures of stereoisomers, and ♦Mitogen-activated protein kinase kinase inhibitors (MEK inhibitors) and pharmaceutically acceptable salts or solvates thereof.

2. The combination according to claim 1, comprising a compound of formula (I), wherein R1 is selected alone or simultaneously or independently when m>1 from a hydrogen atom, a halogen atom, an optionally substituted alkyl group, a haloalkyl group, a fluoroalkyl group, an optionally substituted alkoxy group, and a hydroxyl group.

3. A combination according to claim 1 or 2, comprising a compound of formula (I), wherein R1 is selected from hydrogen atom and chlorine alone or simultaneously or independently when m>1.

4. A combination according to any one of claims 1 to 3, comprising a compound of formula (I), wherein R2 and R3 are simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, an optionally substituted heterocyclic group.

5. A combination according to any one of claims 1 to 4, comprising a compound of formula (I), wherein R2 and R3 are simultaneously or independently selected from a hydrogen atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group and a tert-butyl group.

6. A combination according to any one of claims 1 to 4, comprising a compound of formula (I), wherein R2 and R3 are simultaneously or independently selected from hydrogen and tert-butyl.

7. A combination according to any one of claims 1 to 6, comprising a compound of formula (I), wherein R4 is selected from hydrogen atom, halogen atom, alkyl, haloalkyl, fluoroalkyl, alkoxy, hydroxyl, either alone or simultaneously or independently when n>1.

8. A combination according to any one of claims 1 to 7, comprising a compound of formula (I), wherein R4 is selected alone or simultaneously or independently when n>1 from a hydrogen atom, a hydroxyl group, a methoxy group.

9. The combination according to any one of claims 1 to 8, wherein the compound of formula (I) is selected from the compounds of formula (II), (III) and (IV) below: in • R1 alone or when m>1 is simultaneously or independently selected from hydrogen atom, halogen atom, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted alkoxy, -O-R7, azido, hydroxyl, cyano, nitro, -NR5R6; • R2 and R3 are simultaneously or independently selected from hydrogen atom, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocyclic group; or R2 and R3 can be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group; • R4 is selected alone or simultaneously or independently when n>1 from hydrogen atom, halogen atom, optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted alkoxy, -O-R7, azido, hydroxyl, cyano, nitro, -NR5R6; • R5 and R6 are simultaneously or independently selected from a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted cycloalkyl group, an optionally substituted cycloalkenyl group, an optionally substituted cycloalkynyl group, an optionally substituted heterocyclic group; or R5 and R6 can be linked together with the nitrogen atom to which they are covalently attached to form an optionally substituted heterocyclic group; • R7 may be selected from optionally substituted alkyl, haloalkyl, fluoroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocyclyl; • n is an integer which can have any value of 0, 1, 2, 3 or 4; • m is an integer which may have any of the values ​​of 0, 1, 2, 3, 4 or 5 for formula (II); and m is an integer which may have any of the values ​​of 0, 1, 2, 3 or 4 for formula (III) and (IV); and pharmaceutically acceptable salts, hydrates, solvates, polymorphs, tautomers, isotopic variations, stereoisomers or mixtures of stereoisomers.

10. The combination according to claim 9, wherein for formula (II) or (III) R1, R2, R3 and R4 are as defined in any one of claims 2 to 8, and wherein for formula (IV) R1 and R4 are as defined in any one of claims 2, 3, 7 and 8.

11. The combination according to claim 1, comprising a compound of formula (I), wherein L1 is selected from a single bond and (-CH2-) p Group; R1 is a halogen atom, especially chlorine; R2 and R3 are simultaneously or independently selected from a hydrogen atom and an optionally substituted alkyl group, in particular a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl group; ·m =1， n = 0, and ·p=1， and pharmaceutically acceptable salts, hydrates, solvates, polymorphs, tautomers, isotopic variations, stereoisomers or mixtures of stereoisomers.

12. The combination according to claim 1, wherein the compound of formula (I) is selected from the following compounds: 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (1-5) of formula (Ia) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof.

13. The combination according to claim 1, wherein the compound of formula (I) is selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (1-6) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline hydrochloride (2-3).

14. A combination according to any one of claims 1 to 13, wherein the MEK inhibitor is selected from bemetinib [606143-89-9], cobimetinib [934660-93-2], selumetinib [606143-52-6], trametinib [871700-17-3], AZD8330 [869357-68-6], BI-847325 [1207293-36-4], GDC-0623 [1168091-68-6], Mirdametinib (PD0325901) [391210-10-9], PD184352 (CI-1040) [212631-79-3], Pimasertib (AS-703026) [1236699-92-5], Rifatinib [923032-37-5], TAK-733 [1035555-63-5], BIX 02188 [1094614-84-2], BIX 02189 [1265916-41-3], Honokiol [35354-74-6], Myricetin [529-44-2], PD98059 [167869-21-8], PD318088 [391210-00-7], SL-327 [305350-87-2], U0126 [109511-58-2] or a pharmaceutically acceptable salt or solvate thereof.

15. The combination according to any one of claims 1 to 14, wherein the MEK inhibitor is selected from bemetinib [606143-89-9], cobimetinib [934660-93-2], selumetinib [606143-52-6], trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof.

16. A combination according to any one of claims 1 to 15, comprising 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a salt, solvate thereof and trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof.

17. A combination according to any one of claims 1 to 15, comprising 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a salt or solvate thereof and cobimetinib [934660-93-2] or a pharmaceutically acceptable salt or solvate thereof.

18. A pharmaceutical composition comprising: a therapeutically effective amount of a combination according to any one of claims 1 to 17, wherein the combination comprises a compound of formula (I), (II), (III) or (IV) and a pharmaceutically acceptable salt, hydrate, solvate, polymorph, tautomer, isotopic variant, stereoisomer or mixture of stereoisomers thereof and a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof; and at least one pharmaceutically acceptable carrier, wherein each component of the combination may be contained in a separate pharmaceutical composition or in the same pharmaceutical composition.

19. A combination comprising a compound of formula (I) according to any one of claims 1 to 18 or a pharmaceutically acceptable salt or solvate thereof and a MEK inhibitor or a pharmaceutically acceptable salt or solvate thereof, for use in treating and / or preventing and / or reducing liver cancer-related diseases.

20. A combination for use according to claim 19 comprising a compound of formula (II), (III) or (IV) as defined in claim 9 or 10, or a pharmaceutically acceptable salt or solvate thereof.

21. A combination for use according to claim 19, comprising a compound of formula (I) selected from 2-(4-chlorophenylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (1-5) and 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2).

22. The combination for use according to claim 19, which is a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and trametinib [871700-17-3] or a pharmaceutically acceptable salt or solvate thereof.

23. The combination for use according to claim 19, which is a combination of 2-(4-chlorobenzylamino)-4-(4-tert-butylaminopiperidin-1-yl)-quinoline (2-2) of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof and cobimetinib [934660-93-2] or a pharmaceutically acceptable salt or solvate thereof.

24. The combination for use according to any one of claims 19 to 22, wherein each component of the combination can be used simultaneously, independently or sequentially for liver cancer therapy.

25. The combination for use according to claims 19 to 24, wherein the liver cancer related disease is selected from liver cancer, hepatoblastoma and cholangiocarcinoma.

26. The combination for use according to any one of claims 19 to 25, wherein the liver cancer-related disease is selected from intrahepatic bile duct carcinoma and extrahepatic bile duct carcinoma.

27. The combination for use according to any one of claims 19 to 25, wherein the liver cancer-related disease is liver cancer.

28. A kit comprising a combination of a compound of formula (I) as defined in any one of claims 1 to 18, or a pharmaceutically acceptable salt or solvate thereof, and a MEK inhibitor, or a pharmaceutically acceptable salt or solvate thereof, wherein both components or either component of the combination are in the form of a pharmaceutical composition that can be administered simultaneously, independently or sequentially.

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