Combination therapy for the treatment of hepatocellular carcinoma

Abstract

To provide a combination therapy useful for the treatment of hepatocellular carcinoma.SOLUTION: A method comprises administering the combination of FGFR4 inhibitor, N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide, or a pharmaceutically acceptable salt thereof and Lenvatinib, 4-[3-chloro-4-(N'cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide or a pharmaceutically acceptable salt thereof.SELECTED DRAWING: Figure 1

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 615,848, filed Jan. 10, 2018, which is incorporated by reference as if fully restated herein. FIELD OF THE APPLICATION

[0002] Liver cancer is the second leading cause of mortality from all types of cancer worldwide and the 16th most common cause of death (Llovet JM et al., 2015 “Advances in targeted therapies for hepatocellular carcinoma in the genomic era.” Nat. Rev. Clin Oncology 12, 408-424). Hepatocellular carcinoma (HCC) accounts for up to 90% of all primary liver cancers (Llovet JM et al., 2015).

[0003] Compound 1 is an orally bioavailable selective small molecule FGFR4 inhibitor having the structure shown in Formula I and the chemical name N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide. [ka]

[0004] Compound 1 and its synthesis are reported in WO 2015 / 057938, published on April 23, 2015, which is incorporated herein by reference.

[0005] Lenvatinib (4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide) is a receptor tyrosine kinase (RTK) inhibitor that inhibits the kinase activity of vascular endothelial growth factor (VEGF) receptors VEGFR1 (FLT1), VEGFR2 (KDR), and VEGFR3 (FLT4). Lenvatinib also inhibits other RTKs involved in pathogenic angiogenesis, tumor growth, and cancer progression, as well as normal cell function, including fibroblast growth factor (FGF) receptors FGFR1, 2, 3, and 4; platelet-derived growth factor receptor alpha (PDGFRα), KIT, and RET. Lenvatinib has been reported in U.S. Patent Nos. 7,253,286, 7,612,208, and 9,006,256, all of which are incorporated herein by reference.

[0006] Despite advances in the treatment of HCC, it would be useful to provide alternative treatments for HCC, including treatments for patients whose HCC may be resistant, poorly responsive, or not fully responsive to current treatment regimens.

[0007] An embodiment provides a combination therapy comprising an effective amount of an amount of Compound 1 and an amount of lenvatinib in a patient in need of treatment for hepatocellular carcinoma (HCC). The combination therapy provided herein can result in a synergistic reduction in HCC cell viability and can result in tumor growth inhibition of HCC in the patient in need of treatment. [Brief description of the drawings]

[0008] [Figure 1] 1 is a graph showing the antitumor effect of Compound 1 and Lenvatinib in a Hep3B xenograft model of hepatocellular carcinoma. Both Compound 1 and Lenvatinib were given orally (PO) once daily (QD) for 53 days. Data show the mean ± SEM for tumor volume. [Diagram 2] 1 is a graph showing the results of the PDX xenograft in vivo study reported in Example 2. Detailed Description of the Preferred Embodiments

[0009] Provided herein is a combination therapy useful for treating hepatocellular carcinoma (HCC).In some embodiments, the combination therapy comprises administering compound 1 in combination with lenvatinib.

[0010] Therapeutic drug combinations and methods for administering drug combinations to treat hepatocellular carcinoma are provided herein.As used herein, "therapeutic drug combinations" and similar terms refer to the combination of two therapeutic drugs: (1) Compound 1 and / or the pharmacologically active metabolites, salts, solvates, and racemates of Compound 1, and (2) Lenvatinib and / or the pharmacologically active metabolites, salts, solvates, and racemates of Lenvatinib.Pharmacologically active metabolites include those that are inactive but are converted to pharmacologically active forms in the body after administration.

[0011] Compound 1 is an orally bioavailable, selective, small molecule FGFR4 inhibitor having the structure shown in Formula I. [ka]

[0012] Compound 1 and its synthesis are reported in WO 2015 / 057938, published on April 23, 2015, which is incorporated herein by reference.

[0013] Lenvatinib (4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide) is a receptor tyrosine kinase (RTK) inhibitor that inhibits the kinase activity of vascular endothelial growth factor (VEGF) receptors VEGFR1 (FLT1), VEGFR2 (KDR), and VEGFR3 (FLT4). Lenvatinib also inhibits other RTKs involved in pathogenic angiogenesis, tumor growth, and cancer progression, as well as normal cellular function, including fibroblast growth factor (FGF) receptors FGFR1, 2, 3, and 4; platelet-derived growth factor receptor alpha (PDGFRα), KIT, and RET. Lenvatinib has the following structure: [ka]

[0014] The mesylate salt of lenvatinib, 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide methanesulfonate, also known as LENVIMA® by Eisai, Inc., has the following structure: [ka]

[0015] Lenvatinib is reported in U.S. Pat. Nos. 7,253,286, 7,612,208, and 9,006,256, all of which are incorporated herein by reference.

[0016] The administration of a combination of therapeutic agents includes administration of the individual therapeutic agents of the combination as a single formulation or unitary dosage form, simultaneous but separate administration of the individual therapeutic agents of the combination, or sequential administration of the individual agents of the combination by any suitable route. The administration of the individual therapeutic agents of the combination may also require more frequent administration of one agent compared to the other agent in the combination. Thus, to allow for proper dosing, the packaged pharmaceutical product may include one or more dosage forms that contain the combination of agents, and one or more dosage forms that contain one of the combination of agents but not the other agent of the combination.

[0017] The therapeutic agent may contain one or more asymmetric elements, such as an asymmetric center or axis, e.g., an asymmetric carbon atom, so that the compound may exist as different stereoisomers. Such compounds can be, for example, racemic or optically active. For compounds with two or more asymmetric elements, these compounds can further be mixtures of diastereomers. For compounds with asymmetric centers, it is understood that all optical isomers and mixtures thereof are encompassed. Furthermore, compounds with carbon-carbon double bonds can occur as Z and E forms, and all isomers of the compounds are included in the present invention. In such situations, single enantiomers (optically active forms) can be obtained by asymmetric synthesis, synthesis from optically pure precursors, or by resolution of the racemates. Resolution of the racemates can also be accomplished by conventional methods, such as, for example, crystallization in the presence of a resolving agent, or chromatography, e.g., using a chiral HPLC column.

[0018] The combinations reported herein may include embodiments in which one or more of Compound 1 and Lenvatinib are administered as pharmaceutically acceptable salts or as free bases.Both compounds do not have to be administered as the same pharmaceutically acceptable salts, but may be.In certain embodiments, Compound 1 is administered as a hydrochloride salt and Lenvatinib is administered as a mesylate salt.

[0019] "Pharmaceutically acceptable salts" as used herein refers to acid or base addition salts of the compounds of this disclosure. Pharmaceutically acceptable salts are any salts that retain the activity of the parent compound and do not impart any undue harmful or undesirable effects to the human subject to whom the salt is administered, and in this context, to the subject to whom the salt is administered. Pharmaceutically acceptable salts include, but are not limited to, metal complexes and salts of both inorganic acids and carboxylic acids. Pharmaceutically acceptable salts also include metal salts, such as aluminum, calcium, iron, magnesium, manganese, and complex salts. Further, pharma- ceutically acceptable salts include acetic acid, aspartic acid, alkylsulfonic acid, arylsulfonic acid, axetil, benzenesulfonic acid, benzoic acid, bicarbonate, bisulfate, bitartaric acid, butyric acid, calcium edetate, camsylic acid, carbonic acid, chlorobenzoic acid, citric acid, edetic acid, edisylic acid, estolic acid, esyl, esylic acid, formic acid, fumaric acid, gluceptic acid, gluconic acid, glutamic acid, glycolic acid, glycolylarsanilic acid, hexanmic acid, hexylresorcinoleic acid, hydrabamic acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, hydroxynaphthoic acid, and the like. Non-limiting examples of suitable acid salts include acid salts of isethionic, lactic, lactobionic, maleic, malic, malonic, mandelic, methanesulfonic, methyl nitrate, methyl sulfate, mucic, muconic, napsylic, nitric, oxalic, p-nitromethanesulfonic, pamoic, pantothenic, phosphoric, hydrogen phosphate, dihydrogen phosphate, phthalic, polygalacturonic, propionic, salicylic, stearic, succinic, sulfamic, sulfanic, sulfonic, sulfuric, tannic, tartaric, teoclic, and toluenesulfonic acids.

[0020] An embodiment can include the hydrochloride salt of compound 1. Pharmaceutically acceptable salts can be derived from amino acids, including but not limited to cysteine. Methods for producing compounds as salts are known to those skilled in the art (see, for example, Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Wiley-VCH; Verlag Helvetica Chimica Acta, Zurich, 2002; Berge et al., J. Pharm. Sci. 66: 1, 1977).

[0021] An "effective amount" of a combination of therapeutic agents (eg, Compound 1 and lenvatinib) is an amount sufficient to result in an observable therapeutic benefit compared to baseline HCC.

[0022] The active agents reported herein may be combined with a pharma- ceutically acceptable carrier to produce a pharmaceutical formulation thereof. The particular choice of carrier and formulation will depend on the particular route of administration for which the composition is intended.

[0023] "Pharmaceutically acceptable carrier" as used herein refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound in which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles that may be used in the compositions of the present invention include, but are not limited to, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based materials, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene glycol, and wool fat.

[0024] The compositions of the present invention may be suitable for parenteral, oral, inhalation spray, topical, rectal, nasal, buccal, vaginal or implanted reservoir administration, etc. In some embodiments, the formulation includes ingredients derived from natural or non-natural sources. In some embodiments, the formulation or carrier may be provided in a sterile form. Non-limiting examples of sterile carriers include endotoxin-free water or pyrogen-free water.

[0025] As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In certain embodiments, the compound is administered intravenously, orally, subcutaneously, or by intramuscular administration. The sterile injectable form of the composition of the present invention may be an aqueous or oily suspension. These suspensions may be formulated using suitable dispersing or wetting agents and suspending agents according to techniques known in the art. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution, among others. In addition, sterile, fixed oils are conventionally employed as solvents or suspending media.

[0026] For the above purpose, any bland fixed oil may be used, including synthetic monoglycerides or diglycerides. Fatty acids and their glyceride derivatives are useful for the preparation of injectables, and natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially those polyoxyethylated. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose or similar dispersants commonly used in the preparation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tweens, spans, and other emulsifiers commonly used in the manufacture of pharmaceutically acceptable solid dosage forms, liquid dosage forms, or other dosage forms, may also be used for the formulation.

[0027] For oral administration, the compound or salt may be provided in any acceptable oral dosage form, including, but not limited to, capsules, tablets, aqueous suspensions or solutions. For tablets for oral use, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate may also be added. Useful diluents for oral administration in capsule form include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient may be combined with emulsifying and suspending agents. If necessary, certain sweeteners, flavorings or colorings may also be added. Preservatives may also be added. Suitable examples of pharmaceutically acceptable preservatives include, but are not limited to, various antibacterial and antifungal agents, such as solvents, such as ethanol, propylene glycol, benzyl alcohol, chlorobutanol, quaternary ammonium salts, and parabens (such as methylparaben, ethylparaben, propylparaben, etc.).

[0028] "Immediate release" is meant to include conventional release, where the release of drug begins immediately after administration. As used herein, the term "immediate release" includes dosage forms that allow the drug to dissolve in the gastrointestinal contents without the intention of delaying or prolonging the dissolution or absorption of the drug. The objective is that the drug is released quickly after administration, for example, the drug can release at least 80% of the drug within approximately 30 minutes after the onset of dissolution in a dissolution test.

[0029] "Sustained release" or "sustained release" includes dosage forms in which the time and / or location of drug release characteristics are selected to achieve therapeutic or convenience objectives not provided by conventional dosage forms, such as solutions or immediate release dosage forms.

[0030] The term "steady state" means that a plasma level for a given active agent or combination of active agents is achieved and maintained at a level that is equal to or greater than the minimum effective therapeutic level and below the minimum toxic plasma level for a given active agent(s) upon continuous administration of the active agent(s).

[0031] The term "single formulation" as used herein refers to a single carrier or vehicle formulated to deliver effective amounts of both therapeutic agents to a patient. The single vehicle is designed to deliver an effective amount of each agent along with any pharma- ceutically acceptable carriers or excipients. In some embodiments, the vehicle is a tablet, capsule, pill, or patch.

[0032] The term "unit dose" is used herein to mean that both drugs are administered together in one dosage form to the patient being treated at the same time. In some embodiments, the unit dose is a single formulation. In certain embodiments, the unit dose comprises one or more vehicles, each of which comprises an effective amount of at least one drug (levantinib or compound 1) together with pharma- ceutically acceptable carriers and excipients. In some embodiments, the unit dose is one or more tablets, capsules, pills, or patches administered to the patient at the same time.

[0033] The term "dosage range" as used herein refers to the upper and lower limits of acceptable variation in the amount of a specified drug. Generally, any amount of the drug dose within the specified range can be administered to a patient undergoing treatment.

[0034] The term "treat" is used herein to mean to relieve, reduce or alleviate at least one symptom of a disease in a subject. For example, with respect to HCC, the term "treat" can also mean to prevent, delay the onset (i.e., the period before the clinical pathology of the disease or the symptoms of the disease) and / or to reduce the risk of the onset or worsening of the symptoms of the disease. The term "protect" is used herein to mean to prevent, delay or treat the onset or continuation or worsening of the symptoms of the disease in a subject, or all as appropriate.

[0035] The term "subject" is intended to include animals that may have or are afflicted with HCC. Examples of subjects include mammals, such as humans, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non-human animals. In certain embodiments, the subject is a human, such as a human that has, is at risk of having, or may possibly have HCC.

[0036] The term "about" or "approximately" generally means within 20%, more preferably within 10%, and most preferably even within 5% of a given value or range. Alternatively, particularly in biological systems, the term "about" can mean within one log (i.e., one order of magnitude), and preferably within two-fold of a given value.

[0037] The use of the terms "a" and "an" and "the" and the like in the context of describing the present invention (particularly in the context of the appended claims) should be construed to include both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprising," "having," "including," and "containing" should be construed as open-ended (i.e., meaning "including, but not limited to"), unless otherwise indicated. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring to each individual separate value falling within that range, unless otherwise indicated herein, and each individual value is incorporated into the specification as if it were individually recited herein.

[0038] The term "synergistic effect" as used herein refers to the action of two drugs, such as compound 1 and lenvatinib, that produces an effect, for example, an effect of slowing the progression of HCC, that is greater than the simple sum of the effects of each drug administered by itself.Synergistic effect can be calculated using a suitable method, such as, for example, the sigmoid Emax equation (Holford, NHG and Scheiner, LB, Clin. Pharmacokinet. 6:429-453 (1981)), the Loewe additivity equation (Loewe, S, and Muischnek, H., Arch. Exp. Pathol Pharmacol. 114:313-326 (1926)) and the median effect equation (Chou, TC and Talalay, P., Adv. Enzyme Regul, 22:27-55 (1984)). Each of the equations referred to above can be applied to experimental data to generate corresponding graphs that are useful in evaluating the effect of drug combinations. The corresponding graphs associated with the equations referred to above are the concentration-effect curve, the isobologram curve, and the combination coefficient curve, respectively.

[0039] In some embodiments, treatment is provided for subjects with hepatocellular carcinoma with altered FGFR4 and / or FGF19 (fibroblast growth factor 19) status.

[0040] In some embodiments, the treatment may include or may be performed in conjunction with a step of analyzing FGFR4 and / or FGF19 status in a biological sample containing cells of the hepatocellular carcinoma, and if the hepatocellular carcinoma shows alterations in FGFR4 and / or FGF19, treating the subject with a therapeutically effective amount of a therapeutic combination described herein.

[0041] In order that the embodiments described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are illustrative only and are not to be construed as limiting.

[0042] Treatment methods

[0043] Provided herein are combination therapies useful for treating HCC. As described below, the combinations provided herein may have many advantages.

[0044] One advantage of the combination disclosed herein is the unexpected synergistic effect of the combination of Compound 1 and lenvatinib for the treatment of tumor growth inhibition and for the treatment of HCC.

[0045] In some embodiments, a single pharmaceutical formulation is provided herein that includes the combination of Compound 1 and lenvatinib.The advantage provided herein is the synergistic effect of treating HCC compared with the treatment of a single dose of either drug.When the drug is provided as a single unit dose or a single formulation, the "pill burden" for patients suffering from HCC is not increased.

[0046] As noted above, in one aspect, provided herein is a drug combination useful for treating, preventing, inhibiting, delaying the onset of and / or reducing the risk of developing HCC or reversing HCC in a mammal, comprising administering to the mammal a combination therapy comprising compound 1 and lenvatinib together in effective amounts.

[0047] In some embodiments, the subject (e.g., patient) being treated has been determined to be non-responsive or resistant to one or more HCC therapies, e.g., Compound 1. In other embodiments, the individual being treated responds to Compound 1 therapy, but Compound 1 therapy is improved by administration of lenvatinib. For example, the patient is administered Compound 1 (for some period of time, e.g., more than 1 day, more than 2 days, more than 3 days, more than 1 week, more than 1 month, etc., e.g., 50 mg to 600 mg per day, 200 mg to 400 mg per day, or 300 mg per day). After that time, lenvatinib will be administered to the patient in combination with Compound 1.

[0048] The amount of lenvatinib may vary. For example, lenvatinib may be administered in an amount of 8 mg / day or 12 mg / day. The daily dose may be part of an intermittent dosing regimen lasting 14-21 days or more. The daily dose may be administered as a single dose or as multiple doses.

[0049] Those skilled in the art will recognize that the effective amount of active drug may be lower than the actual amount administered. Thus, dosages necessary to achieve a therapeutic dose are provided herein.

[0050] In various embodiments, a method for treating HCC is provided herein by administering an effective amount of compound 1 and lenvatinib to an individual with HCC. The amount of the drug combination is effective for treating HCC. In one embodiment, the drug combination has a synergistic effect. In one embodiment, even if one or more drugs administered alone at a particular dosage may be effective, when administered in combination, the same dosage of each drug is more effective for treatment. For example, in one embodiment, the combination of compound 1 and lenvatinib is more effective than administration of either drug alone.

[0051] Dosage

[0052] The optimal dosage of a drug combination for the treatment of HCC can be empirically determined for each individual using known methods and will depend on a variety of factors, including the activity of the drugs; the age, weight, general health, sex, and diet of the individual; the time and route of administration; and other drugs the individual is taking. The optimal dosage can be ascertained using routine tests and procedures well known in the art.

[0053] For the combination therapy of the present invention, the daily dose of compound 1 ranges from 50 mg to 600 mg. In some embodiments, the daily dose of compound 1 is up to 600 mg. In certain embodiments, the daily dose of compound 1 is up to 400 mg. In various embodiments, the daily dose of compound 1 is up to 300 mg. In certain embodiments, the daily dose of compound 1 is 200 mg to 400 mg. In one embodiment, the daily dose is 300 mg.

[0054] In certain embodiments, the daily dose of lenvatinib is 8 mg. In certain embodiments, the daily dose of lenvatinib is 12 mg. In certain embodiments, the daily dose of lenvatinib is between 7 mg and 13 mg. In certain embodiments, the daily dose of lenvatinib is between 8 mg and 12 mg.

[0055] The time of administration can be selected so that both drugs are administered simultaneously, separately or consecutively, either in the morning or in the evening. Alternatively, one drug may be administered in the morning and the other in the evening. In certain embodiments, both drugs may be administered once a day, either in the morning or in the evening, as a single tablet, capsule, pill, patch or jelly formulation.

[0056] The amounts of a combination of drugs that may be combined with a carrier material to produce a single dosage form will vary depending on the individual being treated and the particular mode of administration. In some embodiments, a unit dosage form containing a combination of drugs described herein will contain the amount of each drug of the combination that would generally be administered when that drug was administered alone.

[0057] Pharmaceutical Formulations and Routes of Administration

[0058] Provided herein is a pharmaceutical formulation comprising a combination of drugs for the treatment of HCC. The pharmaceutical formulation may further comprise a carrier or excipient, a stabilizer, a flavoring agent, and / or a coloring agent.

[0059] The drug combination may be administered using various routes of administration known to those skilled in the art. Routes of administration include oral administration. In certain embodiments, the pharmaceutical formulation comprising the drug combination may be taken orally in the form of a liquid, syrup, tablet, capsule, powder, sprinkle, chewable tablet, or dissolving disk. Alternatively, the pharmaceutical formulation of the present invention may be administered intravenously or transdermally. Additional routes of administration are known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences, Gennaro AR, Ed., 20th Edition, Mack Publishing Co., Easton, Pa.).

[0060] In some embodiments, compound 1 and lenvatinib are formulated as a paste, jelly, or suspension. For example, the drug is dissolved, encapsulated, or suspended in the form of drug particles, microencapsulated particles, or drug-polymer particles in a gelatinous solution or semisolid. The advantage of an oral jelly formulation is that it is easier to administer the drug to patients who have difficulty swallowing tablets, capsules, or pills. In certain embodiments, both drugs are thoroughly mixed and suspended in a suitable vehicle to form a paste or gel. Additional drugs may be optionally mixed to provide flavor during oral administration. Alginate flavored with peanut butter or raspberry and sweeteners are examples of many suitable taste masking agents. In various embodiments, the paste or jelly may also be formulated with suitable binders or excipients known in the art for topical administration.

[0061] Methods for preparing sustained release formulations in the form of tablets, capsules or pills are known in the art.In some embodiments, sustained release formulations are prepared by coating the active ingredient of a drug with a polymer, preferably a water-insoluble polymer.For example, the water-insoluble polymers used in the pharmaceutical field as sustained release coating, enteric coating, or gastric coating.The water-insoluble polymers can include, for example, ethyl cellulose, purified shellac, white shellac, aminoalkyl methacrylate copolymer RS, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, carboxymethyl ethyl cellulose, cellulose acetate phthalate, methacrylic acid copolymer L, methacrylic acid copolymer LD, methacrylic acid copolymer S, aminoalkyl methacrylate copolymer E, or polyvinyl acetal diethyl amino acetate.

[0062] The type, degree of substitution and molecular weight of the water-insoluble polymer may depend on the solubility of the active ingredient in water or alcohol, the desired sustained release level, etc. The water-insoluble polymers may be used either alone or in combination. Hydrogenated oils, stearic acid, or cetanol as coating aids, and medium chain triglycerides, triacetin, triethyl citrate, or cetanol as plasticizers may further be incorporated.

[0063] In some embodiments, the sustained release formulation is a matrix tablet or granule. The active ingredient may be coated with up to three different types of polymers. These three different types of polymers may include: 1) water-insoluble polymers, such as ethylcellulose; 2) pH-independent gelling polymers, such as hydroxypropylmethylcellulose; and 3) pH-dependent gelling polymers, such as sodium alginate. These three different types of polymers may be used together to reduce the release rate of the drug.

[0064] Dosage form: Release characteristics

[0065] Sustained release formulations can provide a degree of sustained effect, although the exposure and / or bioavailability of the active ingredient may vary based on a variety of factors, including, for example, the absorption window, the carriers or excipients used in the formulation, the mode of delivery of the formulation, and / or the transit time of the active ingredient through the patient's gastrointestinal tract.

[0066] The combination therapy may comprise at least one sustained release portion for performing sustained release function and one immediate release portion for performing immediate release function.In certain embodiments, when the combination therapy is a single dosage form, the dosage form may be in the form of a tablet formed from a mixture of sustained release granules constituting the sustained release portion and immediate release granules constituting the immediate release portion, a capsule preparation obtained by filling a capsule with sustained release granules and immediate release granules, or a dry-coated tablet in which an outer layer constituting the immediate release portion is formed on an inner core constituting the sustained release portion.However, the above embodiments are not limited thereto.

[0067] Furthermore, there are no particular limitations on the state of encapsulation of each drug in the composition or the immediate release portion or the sustained release portion, and Compound 1 may be uniformly dispersed in the composition, the immediate release portion or the sustained release portion, or may be contained in only a portion of the composition, the immediate release portion or the sustained release portion, or may be contained in a concentration gradient.

[0068] The sustained release portion in the composition according to the present invention may contain at least one non-pH-dependent or pH-dependent polymeric substance for controlling drug release.

[0069] The non-pH-dependent polymeric substance used in the present specification may include a polymeric substance whose charge state hardly changes under pH conditions generally found in the gastrointestinal tract, specifically under pH 1 to pH 8. This means, for example, a polymeric substance that does not have a functional group whose charge state changes depending on pH, such as a basic functional group such as an amino group or an acidic functional group such as a carboxylic acid group. The non-pH-dependent polymeric substance may be included in the composition according to the present invention to impart a sustained release function, but may also be included for another purpose. Furthermore, the non-pH-dependent polymeric substance used in the present invention may be water-insoluble, or may swell in water or dissolve in water to form a gel.

[0070] Examples of water insoluble, non-pH dependent polymeric substances include, but are not limited to, cellulose ethers, cellulose esters and methacrylic acid-acrylic acid copolymers (trade name Eudragit, manufactured by Rohm GmbH & Co. KG, Darmstadt, Germany). Examples include, but are not limited to, cellulose alkyl ethers such as ethyl cellulose (trade name Ethocel manufactured by Dow Chemical Company, USA), ethyl methyl cellulose, ethyl propyl cellulose or isopropyl cellulose, and butyl cellulose, cellulose aralkyl ethers such as benzyl cellulose, cellulose cyanoalkyl ethers such as cyanoethyl cellulose, cellulose organic acid esters such as cellulose acetate butyrate, cellulose acetate, cellulose propionate or cellulose butyrate, and cellulose acetate propionate, ethyl acrylate-methyl methacrylate copolymer (trade name Eudragit NE manufactured by ROHM Co., Ltd., Darmstadt, Germany), and aminoalkyl methacrylate copolymer RS ​​(trade names Eudragit RL, Eudragit RS). There is no particular limitation on the average particle size of the water-insoluble polymer used in the present invention, but generally the smaller the average particle size, the better the performance. The average particle size is preferably 0.1 to 100 μm, more preferably 1 to 50 μm, particularly preferably 3 to 15 μm, and most preferably 5 to 15 μm.Further examples of water-soluble or water-swellable non-pH dependent polymeric materials include, but are not limited to, polyethylene oxide (trade name Polyox, molecular weight 100,000-7,000,000, manufactured by Dow Chemical), low substituted hydroxypropyl cellulose (trade name L-HPC, manufactured by Shin-Etsu Chemical, Japan), hydroxypropyl cellulose (trade name HPC, manufactured by Nippon Soda, Co., Ltd, Japan), hydroxypropyl methylcellulose (trade names Metolose 60SH, 65SH, 90SH, manufactured by Shin-Etsu Chemical, Japan), and methylcellulose (trade name Metolose SM, manufactured by Shin-Etsu Chemical, Japan).

[0071] In some embodiments, one non-pH-dependent polymeric substance may be included in the composition, or multiple non-pH-dependent polymeric substances may be included. The non-pH-dependent polymeric substance, when used in the embodiments reported herein, may be a water-insoluble polymeric substance, more preferably ethyl cellulose, ethyl acrylate-methyl methacrylate copolymer (trade name Eudragit NE), or aminoalkyl methacrylate copolymer RS ​​(trade name Eudragit RL, Eudragit RS). At least one of ethyl cellulose and aminoalkyl methacrylate copolymer RS ​​is particularly preferred. Ethyl cellulose is most preferred. There is no particular limit to the amount of non-pH-dependent polymeric substance included in the composition, and this amount can be adjusted as necessary according to the purpose, such as controlling sustained drug release.

[0072] The pH-dependent polymeric substance that can be used in the embodiments reported herein may be a polymeric substance whose charge state changes under pH conditions commonly found in the gastrointestinal tract, specifically pH 1 to pH 8. This means, for example, a polymeric substance having a functional group whose charge state changes depending on the pH, such as a basic functional group such as an amino group or an acidic functional group such as a carboxylic acid group. The pH-dependent functional group of the pH-dependent polymeric substance is preferably an acidic functional group, and most preferably a pH-dependent polymeric substance having a carboxylic acid group.

[0073] The pH-dependent polymeric substances used in the present invention may be water-insoluble or may swell in water or dissolve in water to form a gel. Examples of pH-dependent polymeric substances used in the present invention include, but are not limited to, enteric polymeric substances. Examples of enteric polymeric substances include, but are not limited to, methacrylic acid-methyl methacrylate copolymer (Eudragit L100, Eudragit S100 manufactured by Rohm Co., Ltd., Darmstadt, Germany), methacrylic acid-ethyl acrylate copolymer (Eudragit L100-55, Eudragit L30D-55 manufactured by Rohm Co., Ltd., Darmstadt, Germany), hydroxypropyl methylcellulose phthalate (HP-55, HP-50 manufactured by Shin-Etsu Chemical Co., Ltd., Japan), hydroxypropyl methylcellulose acetate succinate (AQOAT manufactured by Shin-Etsu Chemical Co., Ltd., Japan), carboxymethylethylcellulose (CMEC manufactured by Freund Corporation, Japan), and cellulose acetate phthalate.

[0074] Examples of pH-dependent polymeric substances that swell in water or dissolve in water to form a gel include, but are not limited to, alginic acid, pectin, carboxyvinyl polymer, and carboxymethylcellulose. In the present invention, one pH-dependent polymeric substance may be included in the composition, or multiple pH-dependent polymeric substances may be included. The pH-dependent polymeric substance used in the present invention is preferably an enteric polymeric substance, more preferably methacrylic acid-ethyl acrylate copolymer, methacrylic acid-methyl methacrylate copolymer, hydroxypropyl methylcellulose phthalate, or hydroxypropyl methylcellulose acetate succinate, and particularly preferably methacrylic acid-ethyl acrylate copolymer.

[0075] When a pH-dependent polymeric substance is used in the manufacturing process of the composition according to the present invention, a commercially available product in the form of a powder or granule type, or a suspension type in which the pH-dependent polymeric substance is previously dispersed in a solvent, may be used as is, or such a commercially available product may be dispersed in water or an organic solvent. The smaller the particle size of the pH-dependent polymeric substance, the better it works, and the pH-dependent polymeric substance is preferably in the form of a powder. In the case of methacrylic acid-ethyl acrylate copolymer, an example is Eudragit L100-55. There is no particular limit to the average particle size of the pH-dependent polymeric substance used in the present invention, but the average particle size is preferably 0.05-100 μm, more preferably 0.05-70 μm, and most preferably 0.05-50 μm. Furthermore, there is no particular limit to the amount of the pH-dependent polymeric substance, and for example, in the case of an enteric polymeric substance, the amount is generally 0.1-90 parts by weight, preferably 1-70 parts by weight, more preferably 5-60 parts by weight, and particularly preferably 10-50 parts by weight based on 100 parts by weight of the composition.

[0076] The combination therapy according to the embodiments reported herein may further include any of a variety of additives, such as diluents, lubricants, binders, and disintegrants, as well as any of a variety of pharmacologically acceptable carriers, such as preservatives, colorants, sweeteners, plasticizers, film coating agents, etc. Examples of diluents include, but are not limited to, lactose, mannitol, dibasic calcium phosphate, starch, pregelatinized starch, crystalline cellulose, light anhydrous silicic acid, synthetic aluminum silicate, magnesium aluminate methasilicate, or the like. Examples of lubricants include, but are not limited to, magnesium stearate, calcium stearate, talc, sodium lauryl sulfate, or the like. Examples of binders include, but are not limited to, hydroxypropylcellulose, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, or the like. Examples of disintegrants include, but are not limited to, carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, sodium carboxymethyl starch, low-substituted hydroxypropylcellulose, or the like. Examples of preservatives include, but are not limited to, paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, or the like. Preferred examples of colorants include, but are not limited to, water-insoluble lake colors, natural pigments (e.g., beta-carotene, chlorophyll, red iron oxide), yellow iron oxide, red iron oxide, black iron oxide, or the like. Preferred examples of sweeteners include, but are not limited to, saccharin sodium, dipotassium glycyrrhizinate, aspartame, stevia, or the like. Examples of plasticizers include, but are not limited to, glycerin fatty acid esters, triethyl citrate, propylene glycol, polyethylene glycol, or the like.Examples of film coating agents include, but are not limited to, hydroxypropyl methylcellulose, hydroxypropyl cellulose, or the like.

[0077] Manufacturing method

[0078] One or a combination of conventional methods may be used to manufacture the embodiments reported herein.For example, when manufacturing drug-containing granules as sustained release or immediate release parts, granulation is the main operation, but granulation may be combined with other operations such as mixing, drying, sieving, and classification.As granulation method, for example, wet granulation method in which binder and solvent are added to powder and granulation is performed, dry granulation method in which powder is compressed and granulation is performed, melt granulation method in which binder that melts when heated is added and heated and granulation is performed, or the like may be used.

[0079] Furthermore, according to the granulation method, operation methods such as mixer granulation using a planetary gear mixer, a screw mixer or the like, high-speed mixer granulation using a Henschel mixer, a Spar mixer or the like, extrusion granulation using a cylinder granulator, a rotary granulator, a screw extrusion granulator, a pellet mill granulator or the like, wet high shear granulation, fluidized bed granulation, compression granulation, grinding granulation, or spray granulation can be used. After granulation, drying using a dryer, a fluidized bed or the like, grinding, and sieving can be performed to obtain granules or fine granules for use. Furthermore, a granulation solvent can be used when preparing the composition according to the present invention. There is no specific limitation to such a granulation solvent, and it can be water or any of various organic solvents, such as water, lower alcohols, such as methanol or ethanol, ketones, such as acetone or methyl ethyl ketone, methylene chloride, or mixtures thereof.

[0080] For the sustained release granules included in the embodiment, at least one drug and at least one selected from a non-pH-dependent polymeric substance and a pH-dependent polymeric substance are mixed together, and a diluent and a binder are added as necessary, and granulation is performed to obtain a granular material. The obtained granular material can be dried using a tray dryer, a fluidized bed dryer, or the like, and sieved using a mill or an oscillator, thereby obtaining a sustained release granule. Alternatively, as a method for producing the sustained release granules in the present invention, at least one drug, at least one selected from a non-pH-dependent polymeric substance and a pH-dependent polymeric substance, and a diluent and a binder as necessary can be added using a dry compactor, such as a roller compactor or a slug tablet press, and compressed and mixed at the same time, followed by granulation by crushing to a suitable size. The granular material prepared using such a granulator can be used as it is as a granule or fine granule according to the present invention, or can be further crushed using a power mill, a roll granulator, a rotor speed mill, or the like, and sieved to obtain a sustained release granule. Immediate release granules can be prepared in the same manner as sustained release granules.

[0081] Compression molded product can be manufactured by one or a combination of conventional methods, as drug-containing sustained release part or immediate release part, or as the composition reported herein.For example, at least one drug, at least one selected from non-pH-dependent polymeric substance and pH-dependent polymeric substance, diluent such as mannitol or lactose, binder such as polyvinylpyrrolidone or crystalline cellulose, disintegrant such as carmellose sodium or crospovidone, and lubricant such as magnesium stearate or talc are used, and tableted by conventional method, thereby obtaining compression molded product.In this case, tableting is the main operation in the method of manufacturing compression molded product, but it can be combined with other operations such as mixing, drying, sugar coating and coating.

[0082] Examples of methods for tableting include, but are not limited to, direct compression molding, in which at least one drug and a pharmacologically acceptable additive are mixed together, and then the mixture is directly compressed into a tablet using a tablet press, and dry granule compression or wet granule compression, in which the sustained release granules or immediate release granules according to the present invention are subjected to compression molding after adding a lubricant or disintegrant as necessary. There is no particular limitation on the tablet press used for compression molding, and for example, a single punch tablet press, a rotary tablet press, or a dry-coated tablet press may be used.

[0083] The drug-containing sustained release granules or immediate release granules or the compressed molded product according to the embodiments herein may be used as a composition in the form of granules or tablets as it is, but may be further processed to produce a composition. For example, the compressed molded product or granules may be provided with a film-coating using a film-based material such as ethyl cellulose, casein, methyl cellulose, hydroxypropyl methyl cellulose, methacrylic acid copolymer L, cellulose acetate phthalate, shellac or the like, or may be provided with a sugar-coating liquid containing saccharose, sugar alcohol, gum arabic powder, talc or the like, thereby producing a film-coated tablet or sugar-coated tablet. One of the solvents in this coating technique may be purified water, but organic solvents such as alcohols, ketones, ethers or chlorinated hydrocarbons, or mixtures thereof, may also be used. For example, ethanol, acetone, methylene chloride or the like may be used as the organic solvent. In addition, the coating device may be a device that is normally used in the coating technology for producing drugs, examples of which include a spray coating device in which coating is performed by spraying a coating liquid or the like, and a rotor fluidized bed granulator for layer formation.

[0084] When preparing capsule preparations, they can be prepared by filling the above-mentioned sustained release granules or immediate release granules or mini tablets into hard gelatin capsules or HPMC capsules using an automatic capsule filling machine. Alternatively, in the case of preparations for administration from a tube or dry syrups to be mixed with water or the like when taken, the above-mentioned sustained release granules or immediate release granules can be mixed with a thickening agent or dispersing agent to disperse these granules, and then the mixture is made into granules or tablets. Furthermore, liquids or jellies can be made using water and a substance selected from dispersing agents, emulsifying agents, thickening agents, preservatives, pH adjusting agents, sweeteners, flavoring agents, aromas, etc. However, there is no limitation to the above with respect to other manufacturing methods.

[0085] In order that the embodiments described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are illustrative only and are not to be construed as limiting. EXAMPLES

[0086] Materials and Methods

[0087] Compounds and Drugs Compound 1 is an orally bioavailable, selective small molecule FGFR4 inhibitor developed by H3 Biomedicine. Lenvatinib is an FDA-approved multi-receptor tyrosine inhibitor. Lenvatinib was administered as the mesylate salt and compound 1 was administered as the free base.

[0088] Cell lines tested

[0089] The cell line used, Hep3B (also called HEP3B.1-7), was obtained from the American Type Culture Collection (ATCC), tested for the absence of mycobacterial contamination, and tested for identity by nine-marker short tandem repeat analysis.

[0090] Hep3B xenograft formation

[0091] For human hepatocellular carcinoma cell line Hep3B xenografts, Hep3B cells were cultured in RPMI1640 medium containing 10% FBS at 37°C in 5% CO2 atmosphere and maintained in exponential growth phase. To harvest, cells were washed with PBS, incubated with 0.25% trypsin·EDTA, and suspended in a 1:1 mixture of RPMI1640 medium containing 10% FBS. 5 × 10 7 To prepare an inoculum at a final concentration of cells / mL, cells were spun down and resuspended in RPMI 1640 and Matrigel (Corning). To generate xenografts, 0.1 mL of the inoculum was injected subcutaneously into the right flank region of approximately 8-week-old BALB / c nu / nu female mice weighing 18-20 g to obtain 5 × 10 6 Final concentration of cells / mouse.

[0092] LIX125 patient-derived xenograft (PDX) formation

[0093] Human primary hepatocyte model LIX125 from ChemPartner (shangpharma.com) was implanted into female severe combined immunodeficiency (SCID) mice. Once tumors developed in the mice, the mice were sacrificed and tumors excised for tumor preservation, histopathological diagnosis, and in vivo efficacy studies were implanted into female NU / NU mice. Solid tumor tissues were cleared of necrotic components, cut into 10-15 mg pieces, and mixed. Three to five pieces were mixed with 15-30 mL of Matrigel and implanted into the flanks of 6-8 week old female NU / NU immunodeficient mice weighing 18-20 g. All primary tumors utilized in this study were passaged 3-5 times in vivo, and the respective tumor tissue architecture was maintained throughout the successive implantations.

[0094] Dosing and measurement of antitumor activity

[0095] The mean tumor volume (TV) was approximately 170 mm 3When the mouse reached a 10 day post-implantation age, mice were selected based on their TV and divided into treatment groups with 8 animals per group. Oral (PO) treatment with compound 1 (300 mg / kg for Hep3B and 500 mg / kg for LIX125) alone or in combination with lenvatinib vehicle (control) or lenvatinib (10 mg / kg or 3 mg / kg) administered once daily (QD). Additional groups included treatment with lenvatinib (10 mg / kg or 3 mg / kg) in combination with compound 1 vehicle. (Lenvatinib vehicle includes the vehicle in which lenvatinib is administered, but does not include any lenvatinib. Similarly, compound 1 vehicle includes the vehicle in which compound 1 is administered, but does not include any compound 1.)

[0096] Before dosing, the dosing volume (0.1 mL / 10 g body weight) was calculated from the body weight (BW) of each individual mouse. Body weights were measured daily, and tumor measurements were performed twice weekly. Mice with a weight loss of >20% compared to the body weight on day 1 or tumors with a longest diameter >2,000 mm were immediately euthanized to prevent any pain or suffering of the animals.

[0097] Tumor volume (TV) was calculated in mm according to the following formula: 3 Calculated in units: TV = length x width 2 x 0.5 Length: maximum diameter of tumor (mm) Width: diameter perpendicular to length (mm). Tumor growth inhibition % (TGI) was calculated according to the following formula: Tumor growth inhibition % (TGI) = [(mean control TV on day X - treatment TV on day X) / (mean control TV on day X)] x 100, where day X is any treatment day. Antitumor effects of treatment, partial regression (PR) and complete regression (CR), stable disease (SD) and progressive disease (PD) were defined as follows: PD: 3 consecutive measurements >120% of starting volume or 3 consecutive measurements higher than best response; SD: 3 consecutive measurements >50% and <120% of starting volume; PR: 3 consecutive measurements <50% of starting volume; CR: 3 consecutive measurements <30mm 3 .

[0098] statistical analysis

[0099] Data show the mean ± SEM for tumor volume (N = 8). Differences in TV between groups were analyzed by unpaired multiple t-test, and significance was determined using the Holm-Sidak method. Statistical analysis was performed using GraphPad Prism version 7 (GraphPad Software, La Jolla, CA).

[0100] result

[0101] Example 1: Hep3B cell xenograft in vivo study

[0102] To determine whether lenvatinib enhances the tumor growth inhibition (TGI) exerted by compound 1, the Hep3B cell line was grown as a xenograft in female nude immunodeficient mice. Tumor-bearing mice were orally treated daily for 53 days with (a) 300 mg / kg compound 1 as a single agent along with lenvatinib vehicle; b) (i) 10 mg / kg lenvatinib as a single agent along with compound 1 vehicle; (b) (ii) 3 mg / kg lenvatinib as a single agent along with compound 1 vehicle; (c) (i) 300 mg / kg compound 1 and 10 mg / kg lenvatinib in combination; or (c) (ii) 300 mg / kg compound 1 and 3 mg / kg lenvatinib in combination. The results are shown in Figure 1.

[0103] Single agent compound 1 at 300 mg / kg resulted in tumor growth inhibition with 81% TGI compared to vehicle control (P<0.00001). Lenvatinib at 10 mg / kg as a single agent inhibited tumor growth with 58% TGI compared to vehicle control (P<0.00005). Lenvatinib at 3 mg / kg as a single agent inhibited tumor growth with 23% TGI compared to vehicle control in this Hep3B model (P>0.05). The combination of compound 1 at 300 mg / kg and lenvatinib at 10 mg / kg resulted in enhanced tumor growth inhibition with 98% TGI compared to the single agent groups (P<0.00001). The combination of compound 1 at 300 mg / kg and lenvatinib at 3 mg / kg also resulted in enhanced tumor growth inhibition with 97% TGI compared to the single agent groups. (P<0.00001). These data indicate that lenvatinib can enhance the antitumor effect of compound 1.

[0104] Treatment with compound 1 as a single agent resulted in 2 / 8 SD and 6 / 8 PD in this model. Treatment with lenvatinib at 10 mg / kg as a single agent resulted in 8 / 8 PD in this model. Treatment with lenvatinib at 3 mg / kg as a single agent resulted in 8 / 8 PD in this model. Treatment with compound 1 in combination with lenvatinib at 10 mg / kg resulted in 4 / 8 CR and 4 / 8 SD in this model. Treatment with compound 1 in combination with lenvatinib at 3 mg / kg resulted in 3 / 8 CR and 5 / 8 SD in this model.

[0105] All combination dose groups were well tolerated based on body weight measurements and routine clinical observations.

[0106] Example 2: PDX xenograft in vivo study

[0107] Cell line maintenance

[0108] To determine whether lenvatinib enhances the tumor growth inhibition (TGI) exerted by compound 1 in the PDX model, fragments of LIX125, a hepatocellular carcinoma patient-derived xenograft (PDX) model, were inoculated subcutaneously into the right flank of NU / NU female mice. LIX125 tumor volumes were 100–300 mm 3 When the tumors were in the control group, the animals were randomized into six different groups with eight animals in each group. Tumor-bearing mice were orally treated daily for 32 days with (a) 500 mg / kg compound 1 as a single agent with lenvatinib vehicle; (b)(i) 10 mg / kg lenvatinib as a single agent with compound 1 vehicle; (b)(ii) 3 mg / kg lenvatinib as a single agent with compound 1 vehicle; (c)(i) 500 mg / kg compound 1 and 10 mg / kg lenvatinib in combination; or (c)(ii) 500 mg / kg compound 1 and 3 mg / kg lenvatinib in combination; (d) compound 1 vehicle and lenvatinib vehicle. The results are shown in Figure 2.

[0109] Lenvatinib as a single agent significantly inhibited tumor growth at the 10 mg / kg dose but not at the 3 mg / kg dose (p<0.0007 and p>0.05, respectively) with 72% and 44% TGI, respectively, compared to vehicle control. Single agent Compound 1 at 500 mg / kg did not significantly inhibit tumor growth at 60% TGI, compared to vehicle control (p>0.05).

[0110] The combination of 500 mg / kg compound 1 and 10 mg / kg lenvatinib resulted in a significant increase in tumor growth inhibition with a TGI of 94% compared to 500 mg / kg compound 1 alone (p<0.005). The combination resulted in SD in 3 of 8 animals, PR in 4 of 8 animals, and PD in 1 of 8 animals. The combination of 500 mg / kg compound 1 and 3 mg / kg lenvatinib did not result in a significant increase in tumor growth inhibition with a TGI of 78% compared to 500 mg / kg compound 1 alone (p>0.05). The combination resulted in SD in 2 of 8 animals and PD in 6 of 8 animals. In general, the combination is well tolerated.

[0111] Consistent with the notion that lenvatinib can enhance the effects of Compound 1 in HCC models, additional testing in additional HCC patient-derived models showed enhancement in 4 of 6 models. All PDX models mentioned herein express FGF19.

Claims

1. 1. A combination of therapeutic agents for hepatocellular carcinoma comprising N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof, and 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide or a pharma- ceutically acceptable salt thereof, wherein the combination comprises the N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof in an effective amount that results in a synergistic reduction in viability of hepatocellular carcinoma.

2. The therapeutic combination of claim 1, wherein the combination comprises N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof in a daily dosage of between 50 mg and 600 mg.

3. The combination of therapeutic agents described in claim 1 or 2, wherein the combination comprises N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof in a daily dosage of between 200 mg and 400 mg.

4. The therapeutic combination of any one of claims 1 to 3, wherein the combination comprises N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof in a daily dosage of 300 mg.

5. The therapeutic combination of any one of claims 1 to 4, wherein the combination comprises 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide or a pharma- ceutically acceptable salt thereof in a daily dosage of 8 mg.

6. The therapeutic combination of any one of claims 1 to 4, wherein the combination comprises 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide or a pharma- ceutically acceptable salt thereof in a daily dosage of 12 mg.

7. 7. The therapeutic combination of any one of claims 1 to 6, wherein the N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof and the 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide or a pharma- ceutically acceptable salt thereof are in separate formulations.

8. 7. The therapeutic combination of any one of claims 1 to 6, wherein said N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof and said 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide or a pharma- ceutically acceptable salt thereof are in a single formulation.

9. 9. The therapeutic combination of any one of claims 1 to 8, wherein said N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide is in free base form.

10. The combination of therapeutic agents according to any one of claims 1 to 8, wherein the pharma- ceutically acceptable salt of N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide is the hydrochloride salt of N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide.

11. 11. The therapeutic combination of any one of claims 1 to 10, wherein said pharma- ceutically acceptable salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide is the mesylate salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide.

12. The method of claim 11, wherein the N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide is a compound having a free base form of N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide.

12. The therapeutic combination of any one of claims 1 to 9 and 11, wherein the phenyl group is 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide, and the pharma- ceutically acceptable salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide is the mesylate salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide.

13. 1. A pharmaceutical formulation for use in the treatment of hepatocellular carcinoma comprising N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof, 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, wherein the pharmaceutical formulation comprises the N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof in an amount effective to result in a synergistic reduction in viability of hepatocellular carcinoma.

14. The pharmaceutical formulation of claim 13, wherein the pharmaceutical formulation comprises the N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide or a pharma- ceutically acceptable salt thereof in a daily dosage of between 50 mg and 600 mg.

15. 15. The pharmaceutical formulation of claim 14 comprising the free base form of N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide.

16. 15. The pharmaceutical formulation of claim 14, wherein the pharma- ceutically acceptable salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide is a mesylate salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide.

17. The pharmaceutical formulation of claim 14, wherein the N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide is N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide in free base form, and the pharma- ceutically acceptable salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide is the mesylate salt of 4-[3-chloro-4-(N'-cyclopropylureido)phenoxy]-7-methoxyquinoline-6-carboxamide.

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