JP7289961B1 - Combination therapy for cancer treatment - Google Patents

Abstract

The present invention provides a combination therapy for cancer with a BRAF inhibitor and a MEK inhibitor, and uses and pharmaceutical compositions thereof. A combination of a BRAF inhibitor and a MEK inhibitor is provided, wherein the BRAF inhibitor is a compound of formula (I): TIFF0007289961000051.tif52170 or a pharmaceutically acceptable salt or solvate thereof. [Selection figure] None

Description

The present invention relates to combinations of BRAF inhibitors and MEK inhibitors, as well as uses and pharmaceutical compositions thereof.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である。 The present invention provides, inter alia, BRAF inhibitors and MEK inhibitors for use in treating cancer, wherein the BRAF inhibitor has the formula (I):

or a pharmaceutically acceptable salt or solvate thereof.

Mutant BRAF is a targetable oncogenic driver and so far three BRAF inhibitors (BRAFi) (vemurafenib, dabrafenib, encorafenib) have reached the market and have shown efficacy in BRAFV600E-positive melanoma. However, rapid acquisition of drug resistance is almost universally observed, and the duration of therapeutic benefit for targeted therapies remains limited.

Moreover, the first-generation BRAF inhibitors that were developed revealed an unexpected 'paradoxical' ability to suppress MAPK signaling in BRAF ^V600E- driven tumors, whereas the same inhibitors also showed an unexpected 'paradoxical' ability to suppress MAPK signaling in BRAF wild-type (WT ) model (N Engl J Med 2012;366:271-273; and British Journal of Cancer 111:640-645 (2014)).

Mechanistic studies on the RAF paradox then showed that oncogenic BRAF ^V600E phosphorylates MEK1/2 in its monomeric cytosolic form, whereas activation of WT BRAF and RAF1 induces plasma membrane translocation, as well as activated RAS ( (Nature Reviews Cancer 14:455-467 (2014). )).

Binding of first-generation BRAF inhibitors, such as vemurafenib, dabrafenib and encorafenib, to WT BRAF or RAF1 protomers resulted in RAF homo- and/or heterodimerization and formation of newly formed RAF dimer membranes. Facilitate meetings quickly. In the dimeric conformation, one RAF protomer allosterically induces a conformational change in the second RAF protomer leading to a kinase active state and, importantly, a conformation unfavorable for inhibitor binding. bring a seat Dimerization induced by drug treatment consequently promotes MEK phosphorylation through catalysis operated by unbound protomers through pathway overactivation.

Tumors rarely escape BRAFi treatment and the mechanism in most cases involves an acquired ability to induce RAF dimerization. This effect can be counteracted by MEK inhibitor (MEKi) combination therapy. However, these agents exhibit a very poor therapeutic index that limits the achievable dose of MEKi in humans. Consequently, resistance to first-generation BRAFi and MEKi combination therapy is still mediated by RAF paradoxical activation. Therefore, the clinical benefit of these combination therapies remains limited.

The present invention relates to new combinations of BRAF inhibitors of formula (I) and MEK inhibitors for use in the treatment of cancer, especially melanoma. The compounds of formula (I) are BRAF inhibitors and have negligible paradoxical activity of the MAPK signaling pathway when compared to the first generation BRAF inhibitors on the market: encorafenib, dabrafenib and vemurafenib (paradox inducers). (paradox breaker). In addition to this property, the compounds of formula (I) also possess very potent brain-penetrating properties, thus providing an urgently needed alternative therapy for the treatment of cancers that have metastasized to the brain. do. The present invention has potent combined activity against BRAF-associated tumors with the potential to overcome the rapidly acquired therapeutic resistance frequently observed in patients treated with first-generation BRAF inhibitors. Disclosed are new combinations for use in treating cancer. The combinations disclosed in the present invention for use in treating cancer exhibit unexpected combined activity well beyond the additive effects of MEKi and BRAFi monotherapy.

(3R)-N-[2-cyano-4-fluoro-3-(3-methyl) in combination with the MEK inhibitor cobimetinib in the cell line A375 BRAF/NRAS when compared to the first-generation BRAF inhibitor -4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1-sulfonamide (referred to herein as compound Ia) inhibits P-ERK. Discloses growth inhibition by compound Ia in combination with the MEK inhibitor cobimetinib in the cell line A375 BRAF/NRAS when compared to the first generation BRAF inhibitor encorafenib. We disclose that the combination of Compound Ia and cobimetinib dramatically and synergistically reduces tumor volume in mice engrafted with the cell line A375 NRAS when compared to monotherapy. Discloses a clear synergistic effect of compound Ia in combination with binimetinib on tumor volume in mice implanted with the cell line A375 NRAS when compared to the first generation BRAF inhibitor encorafenib in combination with binimetinib. 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-5-[[3-fluoro -2-(Methylsulfamoylamino)pyridin-4-yl]methyl]benzamide sodium salt (referred to herein as compound IIa) discloses a clear dose-dependent synergistic effect of compound Ia. Discloses a clear synergistic effect of encorafenib in combination with Compound IIa on growth inhibition in cell line A375.

The term "inhibitor" refers to a compound that competes with, reduces binding, or prevents the binding of a specific ligand to a specific receptor, or reduces or prevents the function of a specific protein. In particular, inhibitors as used therein refer to compounds that target, reduce the activity of, or inhibit the activity of the respective target selected from BRAF and MEK, a particular inhibitor being 1 μM It has an IC50 value of less than, less than 500 nM, less than 200 nM, less than 100 nM, less than 50 nM, less than 25 nM, less than 10 nM, less than 5 nM, less than 2 nM or less than 1 nM. In some embodiments of the invention, the term "BRAF inhibitor" refers to reducing BRAF kinase activity by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about Refers to compounds that reduce by 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99%. In some embodiments of the invention, the term "MEK inhibitor" refers to reducing MEK kinase activity by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about Refers to compounds that reduce by 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99%.

The term "IC50" refers to the concentration of a specified compound required to inhibit 50% of a specified measured activity.

The term "pharmaceutically acceptable salt" means a compound of formula (I) or MEK that retains the biological effectiveness and properties of the free base or free acid, which are not biologically or otherwise undesirable. Refers to salts of inhibitors. These salts are, for example, inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., especially hydrochloric acid, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid. , malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine, etc. can be done. Additionally, these salts can be prepared by adding an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium and magnesium salts and the like. Salts derived from organic bases include 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, polyimine resins and the like. Particular pharmaceutically acceptable salts of compounds of formula (I) are the hydrochloride, methanesulfonate and citrate salts. Specific Pharmacies of [3,4-difluoro-2-(2-fluoro-4-iodoanilino)phenyl]-[3-hydroxy-3-[(2S)-piperidin-2-yl]azetidin-1-yl]methanone Legally acceptable salts are fumarate and succinate, especially hemifumarate and hemisuccinate. Certain pharmaceutically acceptable salts of compounds of formula (II) are alkali metal salts such as lithium, sodium, potassium, cesium and rubidium salts, with sodium and potassium salts being preferred.

The term "solvate" refers to a non-covalent stoichiometric or non-stoichiometric combination of a solvent and a solute. The term "hydrate" refers to a non-covalent stoichiometric or non-stoichiometric combination of water and a solute. For example, the compounds of formula (I) and their pharmaceutically acceptable salts can be prepared in unsolvated form and in solvents with pharmaceutically acceptable solvents such as anisole, dichloromethane, toluene, 1,4-dioxane, water, and the like. May exist in sum form.

The compounds of formula (I) contain one asymmetric center and may exist in the form of optically pure enantiomers or mixtures of enantiomers, eg racemates.

According to the Cahn-Ingold-Prelog rule, an asymmetric carbon atom can be of the 'R' or 'S' configuration.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である。 In one aspect, the invention provides BRAF inhibitors and MEK inhibitors for use in treating cancer, wherein the BRAF inhibitor has formula (I):

or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments of the invention, the compound of formula (I) is a compound according to formula (Ia) below:

In some embodiments of the invention, the compound of formula (I) is a compound according to formula (Ib) below:

Non-limiting examples of MEK inhibitors for use in accordance with the present invention include 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2- (methylsulfamoylamino)pyridin-4-yl]methyl]benzamide, cobimetinib, binimetinib, trametinib, selumetinib, pimasertib, lefametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2 -(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (Cl- 1040) and 3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7 (3H,8H)-diones (TAK-733) are included.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である。 In some embodiments of the invention, the MEK inhibitor has formula (II):

or a pharmaceutically acceptable salt or solvate thereof.

The compounds of formula (II) are orally available MEK inhibitors with potent MEK inhibitory activity and high RAF/MEK complex stabilizing activity. The chemical name of formula (II) is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine-4 -yl]methyl]benzamide.

In some embodiments of the invention, the compound of formula (II) is the sodium salt according to formula (IIa) below:

In some embodiments of the invention, the MEK inhibitor is cobimetinib. Cobimetinib is an orally available, potent and highly selective inhibitor of MEK1 and MEK2, central components of the RAS/RAF pathway. Cobimetinib has the chemical name [3,4-difluoro-2-(2-fluoro-4-iodoanilino)phenyl]-[3-hydroxy-3-[(2S)-piperidin-2-yl]azetidin-1-yl] It has methanone and has the following structure:

Cobimetinib can be prepared according to the methods described in WO2007/044515. Cobimetinib is commercially available and has the following CAS registry number: 934660-93-2.

In some embodiments of the invention, the MEK inhibitor is binimetinib. Binimetinib is an orally available, potent and highly selective inhibitor of MEK1 and MEK2, central components of the RAS/RAF pathway. Binimetinib has the chemical name 5-[(4-bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6-carboxamide has the following structure:

Binimetinib can be prepared according to the methods described in WO2003/077914. Binimetinib is commercially available and has the following CAS registry number: 606143-89-9.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である。 In one aspect, the invention provides BRAF inhibitors and MEK inhibitors for use in treating cancer, wherein the MEK inhibitor has formula (II):

or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments of the invention, the BRAF inhibitor is encorafenib. Encorafenib has the chemical name methyl N-[(2S)-1-[[4-[3-[5-chloro-2-fluoro-3-(methanesulfonamido)phenyl]-1-propan-2-ylpyrazole- 4-yl]pyrimidin-2-yl]amino]propan-2-yl]carbamate with the following structure:

Encorafenib can be prepared according to the methods described in WO2011/025927. Encorafenib is commercially available and has the following CAS Registry Number: 1269440-17-6.

Assay procedure
Materials DMEM phenol red free medium supplemented with L-glutamine was purchased from (Thermo Fisher Scientific). Fetal bovine serum (FBS) was purchased from VWR. Enhanced ERK Phospho-T202/Y204 Kit-10,000 test was purchased from Cisbio catalog number 64AERPEH. A375 was originally obtained from ATCC and archived by the Roche repository. The 384-well microplate was 384-well (with lid, HiBase, small volume catalog 784-080) and was purchased from Greiner Bio-One.

HTRF Assay for P-ERK Determination in A375 Cells A375 is a cell cancer model that expresses the V600E mutated BRAF. ERK1,2 phosphorylation, a terminal member of the phosphorylation cascade of the MAPK pathway, has since been reported as the primary readout of the activation state of the MAPK pathway. Prior to assay, the A375 cell line is maintained in DMEM phenol red-free medium supplemented with 10% fetal bovine serum (FBS). FRET fluorescence signal induced by the selective binding of the two antibodies provided in the mentioned kit (Cisbio catalog number 64AERPEH) against the ERK protein when phosphorylated at the Thr202/Tyr204 positions after compound treatment. P-ERK levels are determined by measuring . Briefly, 8000 cells/well in 12 μl medium/well were seeded into a 384-well plate and left in an incubator overnight (at 37° C. in a humidified atmosphere of 5% CO2), the next day the plates were subjected to the following final Treat in duplicate with test compound, dabrafenib and PLX8394 (the latter two as controls) at drug concentrations: 10 μM-3 μM-1 μM-0.3 μM-0.1 μM-0.03 μM-0.01 μM-0. .003 μM-0.001 μM, all wells subject to DMSO normalization, drug incubation for 1 hour. 4 μl of 4× lysis buffer supplied with the kit is then added to the wells, then the plate is centrifuged (300 rcf) for 30 seconds and incubated for 1 hour at room temperature on a plate shaker.

At the end of the incubation, 4 μL/well of enhanced P-ERK antibody solution (prepared according to manufacturer's instructions) followed by 4 μL/well of Cryptate P-ERK antibody solution (prepared according to manufacturer's instructions) (Cisbio catalog number 64AERPEH). ) is added to the test wells.
To allow for proper data normalization, reported no drug treatment control wells are always included on each plate (according to manufacturer's instructions):
Control and experimental p-ERK HTRF well contents (μl):

Plates are then centrifuged at 300 rcf for 30 seconds, sealed to prevent evaporation, and incubated overnight at room temperature in the dark.

Plates are then analyzed and fluorescence emission values are collected at 665 and 620 nM by a Pherastast FSX (BMG Labtech) instrument.

The fluorescence values obtained are processed according to the formula: ratio=signal(620 nm)/signal(625 nm)*10000, then the average blank ratio is subtracted for all values.

表１：化合物（Ｉａ）および（Ｉｂ）は、ＢＲＡＦおよびＢＲＡＦ^{Ｖ６００Ｅ}に対して高い親和性ならびにＣ末端Ｓｒｃキナーゼ（ＣＳＫ）およびリンパ球特異的チロシンタンパク質キナーゼ（ＬＣＫ）よりも高い選択性を有する。Ｋｄは、生化学実験における解離定数である。ＩＣ５０を、上記のようにＡ３７５細胞株において測定した。 The average of the ratios (blank subtracted) derived by cells treated with DMSO only (blank subtracted) was taken as 100% and the average of ratios (blank subtracted) derived by cells treated with 10 μM dabrafenib was taken as 0%. and normalize the data. The mean of the normalized points is fitted with a sigmoidal curve to determine the IC50. Results are summarized in Table 1. Biochemical assays confirmed the high affinity of compounds of formulas (Ia) and (Ib) for BRAF and BRAF ^V600E . Kd is the dissociation constant in biochemical experiments.

Table 1: Compounds (Ia) and (Ib) have high affinity for BRAF and BRAF ^V600E and high selectivity over C-terminal Src kinase (CSK) and lymphocyte-specific tyrosine protein kinase (LCK). Kd is the dissociation constant in biochemical experiments. IC50 was determined in the A375 cell line as described above.

The preparation of compounds of formula (I) of the present invention may be carried out in sequential or convergent synthetic routes. The syntheses of the present invention are illustrated in the following general schemes. The skills necessary to carry out the reaction and purification of the resulting product are known to those skilled in the art.

In more detail, the compounds of formula (I) can be manufactured by the methods given below, by the methods given in the examples, or by analogous methods. Suitable reaction conditions for the individual reaction steps are known to those skilled in the art. The reaction order is not limited to that displayed in Scheme 1, but depending on the starting materials and their respective reactivity the order of the reaction steps can be freely changed. Starting materials are either commercially available or prepared by methods analogous to those shown below, described in the references cited herein or in the experimental procedures, or known in the art. be able to.

It will be appreciated that the compounds of formula (I) in the present invention may be derivatized at functional groups to provide derivatives that are convertible back to the parent compound in vivo.

２－アミノ－５－ヒドロキシ安息香酸（１０ｇ、６５．３ｍｍｏｌ、当量：１．０）およびＮ－メチルホルムアミド（３０ｇ、２９．９ｍＬ、５０３ｍｍｏｌ、当量：７．７）を１４５℃で２１時間４５分間加熱し、次いで室温に冷却した。反応混合物を５０ｍＬのＨ_２Ｏで希釈し、室温で２０分間撹拌した。得られた沈殿物を濾過によって回収した。淡褐色固体を２０ｍＬの水で３回洗浄した。固体をトルエンに溶解し、蒸発乾固させた（３回）。固体を高真空下、４０℃の真空中で一晩乾燥させて、表題化合物を淡褐色固体（１０．３ｇ、収率８９％）として得た。ＭＳ（ＥＳＩ）ｍ／ｚ：１７７．１［Ｍ＋Ｈ］^＋． Experimental Procedure (3R)-N-[2-Cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1-sulfonamide ( compounds of formula (Ia)) and (3S)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine -1-sulfonamides (compounds of formula (Ib))
6-hydroxy-3-methyl-quinazolin-4-one

2-amino-5-hydroxybenzoic acid (10 g, 65.3 mmol, equivalent weight: 1.0) and N-methylformamide (30 g, 29.9 mL, 503 mmol, equivalent weight: 7.7) at 145° C. for 21 hours and 45 minutes. Heat and then cool to room temperature. The reaction mixture was diluted with 50 mL of H ₂ O and stirred at room temperature for 20 minutes. The resulting precipitate was collected by filtration. The light brown solid was washed with 20 mL of water three times. The solid was dissolved in toluene and evaporated to dryness (three times). The solid was dried under high vacuum at 40° C. overnight to give the title compound as a pale brown solid (10.3 g, 89% yield). MS (ESI) m/z: 177.1 [M+H] ⁺ .

炭酸セシウム（３．２２ｇ、９．７９ｍｍｏｌ、当量：１．１５）を、Ｎ，Ｎ－ジメチルホルムアミド（３５ｍＬ）中の６－ヒドロキシ－３－メチルキナゾリン－４－オン（１５００ｍｇ、８．５１ｍｍｏｌ、当量：１．０）の溶液に室温で添加した。混合物を室温で３０分間撹拌し、次いで２，３，６－トリフルオロベンゾニトリル（１．４７ｇ、１．０８ｍｌ、９．３７ｍｍｏｌ、当量：１．１）を添加した。１時間後、反応物を氷上で冷却し、水（１２０ｍＬ）で希釈した。得られた固体を濾過によって回収し、氷水（１００ｍＬ）およびヘプタン（１００ｍＬ）で洗浄し、吸引乾燥させた。固体をトルエンに溶解し、蒸発乾固させ（３回）、次いで真空中で一晩乾燥させて、表題化合物を淡褐色固体（２．５８ｇ、収率９７％）として得た。ＭＳ（ＥＳＩ）ｍ／ｚ：３１４．１［Ｍ＋Ｈ］＋． 3,6-difluoro-2-(3-methyl-4-oxo-quinazolin-6-yl)oxy-benzonitrile

Cesium carbonate (3.22 g, 9.79 mmol, eq: 1.15) was treated with 6-hydroxy-3-methylquinazolin-4-one (1500 mg, 8.51 mmol, eq: 1.15) in N,N-dimethylformamide (35 mL). : 1.0) at room temperature. The mixture was stirred at room temperature for 30 minutes, then 2,3,6-trifluorobenzonitrile (1.47 g, 1.08 ml, 9.37 mmol, equivalent weight: 1.1) was added. After 1 hour, the reaction was cooled on ice and diluted with water (120 mL). The resulting solid was collected by filtration, washed with ice water (100 mL) and heptane (100 mL), and sucked dry. The solid was dissolved in toluene and evaporated to dryness (three times) then dried in vacuo overnight to give the title compound as a light brown solid (2.58g, 97% yield). MS (ESI) m/z: 314.1 [M+H]+.

（Ｒ）－３－フルオロピロリジン塩酸塩（１．８ｇ、１４．３ｍｍｏｌ、当量：１．２）を、ジオキサン（１０ｍＬ）中の硫酸ジアミド（１．１４８ｇ、１１．９ｍｍｏｌ、当量：１．０）およびトリエチルアミン（２．４２ｇ、３．３３ｍＬ、２３．９ｍｍｏｌ、当量：２）の溶液に添加した。反応物を密閉管中１１５℃で１５．５時間撹拌し、次いで室温に冷却し、真空中で濃縮した。残渣をＤＣＭで希釈し、シリカゲルで蒸発乾固させ、カラムに移した。フラッシュクロマトグラフィー（４０ｇのシリカ、８０％ＥｔＯＡｃ）による精製によって表題化合物を白色結晶性固体（１．８２ｇ、収率９１％）として得た。ＭＳ（ＥＳＩ）ｍ／ｚ：１６９．１［Ｍ＋Ｈ］^＋． (3R)-3-fluoropyrrolidine-1-sulfonamide

(R)-3-fluoropyrrolidine hydrochloride (1.8 g, 14.3 mmol, eq: 1.2) was treated with diamide sulfate (1.148 g, 11.9 mmol, eq: 1.0) in dioxane (10 mL). and triethylamine (2.42 g, 3.33 mL, 23.9 mmol, eq: 2). The reaction was stirred in a sealed tube at 115° C. for 15.5 hours, then cooled to room temperature and concentrated in vacuo. The residue was diluted with DCM, evaporated to dryness with silica gel and transferred to a column. Purification by flash chromatography (40 g silica, 80% EtOAc) gave the title compound as a white crystalline solid (1.82 g, 91% yield). MS (ESI) m/z: 169.1 [M+H] ⁺ .

トリエチルアミン（３０４ｍｇ、４１９μｌ、３．０１ｍｍｏｌ、当量：２．０）を、ジオキサン（１．３ｍｌ）中の硫酸ジアミド（１４６ｍｇ、１．５ｍｍｏｌ、当量：１．０）および（Ｓ）－３－フルオロピロリジン塩酸塩（２３４ｍｇ、１．８ｍｍｏｌ、当量：１．２）の懸濁液に添加した。反応物を密閉管中１１５℃で１６時間３５分間撹拌し、次いで真空中で濃縮した。残渣をＭｅＯＨで希釈し、シリカゲルで蒸発乾固させ、カラムに移した。フラッシュクロマトグラフィー（４０ｇのシリカ、０～８％ＭｅＯＨ／ＤＣＭ）による精製によって表題化合物を淡黄色固体（１９３ｍｇ、収率７５％）として得た。ＭＳ（ＥＳＩ）ｍ／ｚ：１６９．１［Ｍ＋Ｈ］^＋． (3S)-3-fluoropyrrolidine-1-sulfonamide

Triethylamine (304 mg, 419 μl, 3.01 mmol, eq: 2.0) was treated with diamide sulfate (146 mg, 1.5 mmol, eq: 1.0) and (S)-3-fluoropyrrolidine in dioxane (1.3 ml). Added to a suspension of hydrochloride (234 mg, 1.8 mmol, equivalents: 1.2). The reaction was stirred in a sealed tube at 115° C. for 16 hours and 35 minutes, then concentrated in vacuo. The residue was diluted with MeOH, evaporated to dryness over silica gel and transferred to a column. Purification by flash chromatography (40 g silica, 0-8% MeOH/DCM) gave the title compound as a pale yellow solid (193 mg, 75% yield). MS (ESI) m/z: 169.1 [M+H] ⁺ .

（Ｒ）－３－フルオロピロリジン－１－スルホンアミド（１．２６ｇ、７．５１ｍｍｏｌ、当量：２．１）および炭酸セシウム（２．５６ｇ、７．８７ｍｍｏｌ、当量：２．２）を、アルゴン雰囲気下、乾燥ＤＭＦ（１０．２ｍｌ）に懸濁した。反応物を５０℃で３０分間撹拌した。反応混合物を室温に冷却し、ＤＭＦ（２５．５ｍｌ）中の３，６－ジフルオロ－２－（（３－メチル－４－オキソ－３，４－ジヒドロキナゾリン－６－イル）オキシ）ベンゾニトリル（１．１２ｇ、３．５８ｍｍｏｌ、当量：１．０）の溶液を添加した。反応混合物を１００℃で１５時間撹拌し、次いで真空中で濃縮した。残渣を飽和ＮＨ_４Ｃｌ（１００ｍＬ）水溶液およびＥｔＯＡｃ（１００ｍＬ）に溶解した。相を分離し、水層を２×１００ｍＬのＥｔＯＡｃでさらに抽出した。合わせた有機層を水（２００ｍＬ）およびブライン（２００ｍＬ）で洗浄し、乾燥させ（Ｎａ_２ＳＯ_４）、フィルタにかけ、真空中で濃縮した。水層をＥｔＯＡｃ（３×１００ｍＬ）で逆抽出した。合わせた有機抽出物をブライン（２００ｍＬ）で洗浄し、乾燥させ（Ｎａ_２ＳＯ_４）、フィルタにかけ、真空中で濃縮した。残渣をＤＣＭおよびＭｅＯＨで希釈し、シリカで濃縮した。フラッシュクロマトグラフィー（１２０ｇ、０．５～２％ＭｅＯＨ／ＤＣＭ）による精製によってオフホワイトの固体が得られ、これを超音波処理により１：１ヘプタン／ＤＣＭ（２０ｍＬ）でトリチュエートし、次いで真空中で乾燥させて、表題化合物を無色固体（１．０８７ｇ、収率６６％）として得た。ＭＳ（ＥＳＩ）ｍ／ｚ：４２６．２［Ｍ＋Ｈ］^＋．キラルＳＦＣ：ＲＴ＝４．５９４分［Ｃｈｉｒａｌｐａｋ ＩＣカラム、４．６×２５０ｍｍ、粒径５μｍ（Ｄａｉｃｅｌ）；８分間にわたり０．２％ＮＨＥｔ_２を含有する２０～４０％ＭｅＯＨの勾配；流量：２．５ｍＬ／分；１４０バールの背圧］。 (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1-sulfonamide (formula ( Compounds of Ia))

(R)-3-fluoropyrrolidine-1-sulfonamide (1.26 g, 7.51 mmol, equivalent weight: 2.1) and cesium carbonate (2.56 g, 7.87 mmol, equivalent weight: 2.2) were added in an argon atmosphere. The bottom was suspended in dry DMF (10.2 ml). The reaction was stirred at 50° C. for 30 minutes. The reaction mixture was cooled to room temperature and 3,6-difluoro-2-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)benzonitrile ((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy) in DMF (25.5 ml) A solution of 1.12 g, 3.58 mmol, equivalent weight: 1.0) was added. The reaction mixture was stirred at 100° C. for 15 hours and then concentrated in vacuo. The residue was dissolved in saturated aqueous NH ₄ Cl (100 mL) and EtOAc (100 mL). The phases were separated and the aqueous layer was further extracted with 2 x 100 mL of EtOAc. The combined organic layers were washed with water (200 mL) and brine (200 _mL ), dried ( _Na2SO4 ), filtered and concentrated in vacuo. The aqueous layer was back extracted with EtOAc (3 x 100 mL). _The combined organic extracts were washed with brine (200 mL), dried ( _Na2SO4 ), filtered and concentrated in vacuo. The residue was diluted with DCM and MeOH and concentrated onto silica. Purification by flash chromatography (120 g, 0.5-2% MeOH/DCM) gave an off-white solid, which was triturated with 1:1 heptane/DCM (20 mL) by sonication and then in vacuo. Dried to give the title compound as a colorless solid (1.087 g, 66% yield). MS (ESI) m/z: 426.2 [M+H] ⁺ . Chiral SFC: RT=4.594 min [Chiralpak IC column, 4.6×250 mm, 5 μm particle size (Daicel); gradient of 20-40% MeOH containing 0.2% NHEt ₂ over 8 min; .5 mL/min; 140 bar back pressure].

（Ｓ）－３－フルオロピロリジン－１－スルホンアミド（１８１ｍｇ、１．０８ｍｍｏｌ、当量：２．１）をＤＭＦ（１．６ｍｌ）に溶解した。室温で、炭酸セシウム（３６８ｍｇ、１．１３ｍｍｏｌ、当量：２．２）を添加し、反応混合物を５０℃で３０分間撹拌した。反応混合物を室温に冷却し、ＤＭＦ（４ｍｌ）中の３，６－ジフルオロ－２－（（３－メチル－４－オキソ－３，４－ジヒドロキナゾリン－６－イル）オキシ）ベンゾニトリル（１６０．８ｍｇ、５１３μｍｏｌ、当量：１．０）の溶液を添加した。反応混合物を１０５℃で２時間５０分間撹拌し、次いで真空中で濃縮した。残渣をＤＣＭに溶解し、飽和ＮＨ_４Ｃｌ水溶液で洗浄した。水層をＤＣＭで２回逆抽出した。合わせた有機層をＮａ_２ＳＯ_４で乾燥させ、フィルタにかけ、蒸発させた。残渣（褐色油状物）をＤＣＭで希釈し、カラムに移した。フラッシュクロマトグラフィー（８０ｇ、ＤＣＭ中０～１００％ＥｔＯＡｃ）による精製によって固体が得られ、これをＳＦＣによってさらに精製して、表題化合物を淡黄色固体（１１９ｍｇ、収率５０％）として得た。ＭＳ（ＥＳＩ）ｍ／ｚ：４２６．２［Ｍ＋Ｈ］^＋．キラルＳＦＣ：ＲＴ＝４．４１１分［Ｃｈｉｒａｌｐａｋ ＩＣカラム、４．６×２５０ｍｍ、粒径５μｍ（Ｄａｉｃｅｌ）；８分間にわたり０．２％ＮＨＥｔ_２を含有する２０～４０％ＭｅＯＨの勾配；流量：２．５ｍＬ／分；１４０バールの背圧］。 (3S)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1-sulfonamide (formula ( Compounds of Ib))

(S)-3-fluoropyrrolidine-1-sulfonamide (181 mg, 1.08 mmol, equivalent weight: 2.1) was dissolved in DMF (1.6 ml). At room temperature, cesium carbonate (368 mg, 1.13 mmol, equivalents: 2.2) was added and the reaction mixture was stirred at 50° C. for 30 minutes. The reaction mixture was cooled to room temperature and treated with 3,6-difluoro-2-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)benzonitrile (160. 8 mg, 513 μmol, equiv: 1.0) solution was added. The reaction mixture was stirred at 105° C. for 2 hours 50 minutes and then concentrated in vacuo. The residue was dissolved in DCM and washed with saturated aqueous NH ₄ Cl. The aqueous layer was back extracted twice with DCM. The combined organic layers were dried _{over Na2SO4} , filtered and evaporated. The residue (brown oil) was diluted with DCM and transferred to a column. Purification by flash chromatography (80 g, 0-100% EtOAc in DCM) gave a solid which was further purified by SFC to give the title compound as a pale yellow solid (119 mg, 50% yield). MS (ESI) m/z: 426.2 [M+H] ⁺ . Chiral SFC: RT=4.411 min [Chiralpak IC column, 4.6×250 mm, 5 μm particle size (Daicel); gradient of 20-40% MeOH containing 0.2% NHEt ₂ over 8 min; .5 mL/min; 140 bar back pressure].

2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl]benzamide (formula ( II) compound)
Compounds of formula (II) can be manufactured by the methods given below, by the methods given in the examples or by analogous methods.

NMR analysis was performed by Bruker Co. was performed using an AVANCE III HD400 (400 MHz) from NMR data are reported in ppm (parts per million) (δ) and were referenced to the deuterium lock signal from the sample solvent.

Mass spectral data were obtained from Shimadzu Corp. Single quadrupole mass spectrometer (LCMS-2020) with ultra-performance liquid chromatography (Nexera UC) from Waters Co. were obtained using a single quadrupole mass spectrometer (SQD or SQD2) equipped with an Acquity Ultra Performance Liquid Chromatography (UPLC or UPLC I-Class).

High performance liquid chromatography was performed using one of analytical conditions AC listed in Table 2 below. In Table 2, "TFA" stands for trifluoroacetic acid and "FA" for formic acid.

Commercially available reagents were used directly without further purification.

All non-aqueous reactions were carried out in anhydrous solvents.

Concentration under reduced pressure and solvent distillation were performed using a rotary evaporator.

As used herein, "room temperature" means a temperature of about 20°C to about 25°C.

トルエン（４４ｍＬ）とＭｅＯＨ（１１ｍＬ）中の３，４－ジフルオロ－２－（（２－フルオロ－４－ヨードフェニル）アミノ）－５－ホルミル安息香酸（５．５０ｇ、１３．１ｍｍｏｌ）の混合懸濁液を０℃に冷却し、１０％ジアゾメチルトリメチルシランヘキサン溶液（２１．８ｍＬ、１３．１ｍｍｏｌ）を添加し、混合物を室温で６４時間撹拌した。酢酸（０．７４８ｍＬ）を反応混合物に添加し、次いで減圧下で濃縮した。得られた残渣をトリチュエーション（ヘキサン／酢酸エチル）によって精製して、表題化合物（５．０１ｇ、８８％）を無色固体として得た。ＬＣＭＳ ｍ／ｚ：４３６［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：１．００分（分析条件Ｂ） Methyl 3,4-difluoro-2-(2-fluoro-4-iodoanilino)-5-formylbenzoate (compound a1)

A mixed suspension of 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-formylbenzoic acid (5.50 g, 13.1 mmol) in toluene (44 mL) and MeOH (11 mL). The turbidity was cooled to 0° C., 10% diazomethyltrimethylsilane in hexane (21.8 mL, 13.1 mmol) was added and the mixture was stirred at room temperature for 64 hours. Acetic acid (0.748 mL) was added to the reaction mixture and then concentrated under reduced pressure. The resulting residue was purified by trituration (hexane/ethyl acetate) to give the title compound (5.01 g, 88%) as a colorless solid. LCMS m/z: 436 [M+H] ⁺ . HPLC retention time: 1.00 minutes (analysis condition B)

４－メチルベンゼンスルホニルヒドラジド（２．１４ｇ、１１．５ｍｍｏｌ）を、ＥｔＯＨ（１００ｍＬ）中のメチル３，４－ジフルオロ－２－（２－フルオロ－４－ヨードアニリノ）－５－ホルミルベンゾエート（化合物ａ１、５．００ｇ、１１．５ｍｍｏｌ）の懸濁液に添加し、混合物を室温で３時間撹拌した。反応混合物を減圧下で濃縮し、次いでヘキサン（１５０ｍＬ）を添加した。混合物を０℃に冷却し、フィルタにかけ、次いでヘキサン（３０ｍＬ）で洗浄して、表題化合物（７．０５ｇ、定量的）を固体として得た。ＬＣＭＳ ｍ／ｚ：６０４［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：１．０６分（分析条件Ｂ） Methyl 3,4-difluoro-2-(2-fluoro-4-iodoanilino)-5-[(E)-[(4-methylphenyl)sulfonylhydrazinylidene]methyl]benzoate (compound a2)

4-Methylbenzenesulfonyl hydrazide (2.14 g, 11.5 mmol) was treated with methyl 3,4-difluoro-2-(2-fluoro-4-iodoanilino)-5-formylbenzoate (compound a1, 5.00 g, 11.5 mmol) and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure and then hexane (150 mL) was added. The mixture was cooled to 0° C., filtered, then washed with hexane (30 mL) to give the title compound (7.05 g, quantitative) as a solid. LCMS m/z: 604 [M+H] ⁺ . HPLC retention time: 1.06 minutes (analysis condition B)

トリエチルアミン（３．６３ｍＬ、２６．０ｍｍｏｌ）および１－（２，４－ジメトキシフェニル）メタンアミン（３．２６ｍＬ、２１．７ｍｍｏｌ）を、ＮＭＰ（３２ｍＬ）中の２，３－ジフルオロ－４－ヨードピリジン（２．０９ｇ、８．６７ｍｍｏｌ）の溶液に添加し、混合物を１００℃で１．５時間撹拌した。水を反応混合物に添加し、酢酸エチルで抽出を行った。有機層を１３％ブラインで洗浄し、無水硫酸ナトリウムで乾燥させ、乾燥剤を濾別した後、減圧下で濃縮した。得られた残渣をシリカゲル・カラム・クロマトグラフィー（ヘキサン／酢酸エチル）によって精製して、表題化合物（３．２０ｇ、９５％）をオイル状物として得た。ＬＣＭＳ ｍ／ｚ：３８９［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：０．９４分（分析条件Ｃ） N-(2,4-dimethoxybenzyl)-3-fluoro-4-iodopyridin-2-amine (compound a3)

Triethylamine (3.63 mL, 26.0 mmol) and 1-(2,4-dimethoxyphenyl)methanamine (3.26 mL, 21.7 mmol) were combined with 2,3-difluoro-4-iodopyridine ( 2.09 g, 8.67 mmol) and the mixture was stirred at 100° C. for 1.5 hours. Water was added to the reaction mixture and extraction was performed with ethyl acetate. The organic layer was washed with 13% brine, dried over anhydrous sodium sulfate, filtered off the drying agent, and then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (3.20 g, 95%) as an oil. LCMS m/z: 389 [M+H] ⁺ . HPLC retention time: 0.94 minutes (analysis condition C)

Ｎ－（２，４－ジメトキシベンジル）－３－フルオロ－４－ヨードピリジン－２－アミン（化合物ａ３、２．７０ｇ、６．９６ｍｍｏｌ）、［１，１－ビス（ジフェニルホスフィノ）フェロセン］ジクロロパラジウム（ＩＩ）ジクロロメタン付加物（５６８ｍｇ、０．６９６ｍｍｏｌ）、酢酸カリウム（２．０５ｇ、２０．９ｍｍｏｌ）およびビス（ピナコラト）ジボロン（２．６５ｇ、１０．４ｍｍｏｌ）の１，４－ジオキサン溶液（２７ｍＬ）を、窒素雰囲気下、９０℃で５時間、次いで１１０℃で１９時間撹拌した。反応混合物を減圧下で濃縮し、得られた残渣を逆相カラムクロマトグラフィー（０．１％ギ酸水溶液／０．１％ギ酸アセトニトリル溶液）によって精製して、表題化合物（２．０７ｇ、９７％）をオイル状物として得た。ＬＣＭＳ ｍ／ｚ：３０７［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：０．４４分（分析条件Ｃ） [2-[(2,4-dimethoxyphenyl)methylamino]-3-fluoropyridin-4-yl]boronic acid (compound a4)

N-(2,4-dimethoxybenzyl)-3-fluoro-4-iodopyridin-2-amine (compound a3, 2.70 g, 6.96 mmol), [1,1-bis(diphenylphosphino)ferrocene]dichloro Palladium(II) dichloromethane adduct (568 mg, 0.696 mmol), potassium acetate (2.05 g, 20.9 mmol) and bis(pinacolato)diboron (2.65 g, 10.4 mmol) in 1,4-dioxane (27 mL) ) was stirred at 90° C. for 5 hours and then at 110° C. for 19 hours under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by reverse phase column chromatography (0.1% formic acid in water/0.1% formic acid in acetonitrile) to give the title compound (2.07 g, 97%). was obtained as an oil. LCMS m/z: 307 [M+H] ⁺ . HPLC retention time: 0.44 minutes (analysis condition C)

メチル３，４－ジフルオロ－２－（２－フルオロ－４－ヨードアニリノ）－５－［（Ｅ）－［（４－メチルフェニル）スルホニルヒドラジニリデン］メチル］ベンゾエート（化合物ａ２、１．３０ｇ、２．１６ｍｍｏｌ）、［２－［（２，４－ジメトキシフェニル）メチルアミノ］－３－フルオロピリジン－４－イル］ボロン酸（化合物ａ４、１．９８ｇ、６．４６ｍｍｏｌ）および炭酸カリウム（３５７ｍｇ、２．５９ｍｍｏｌ）の１，４－ジオキサン懸濁液（５９ｍＬ）を、窒素雰囲気下、１００℃で２．５時間、次いで１１０℃で３時間撹拌した。酢酸エチルを反応混合物に添加し、次いで水および１３％ブラインで洗浄した。有機層を無水硫酸ナトリウムで乾燥させ、乾燥剤を濾別した後、減圧下で濃縮した。得られた残渣をシリカゲル・カラム・クロマトグラフィー（ヘキサン／酢酸エチル）によって精製して、表題化合物（５２４ｍｇ、３６％）を泡状物として得た。ＬＣＭＳ ｍ／ｚ：６８２［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：１．０３分（分析条件Ｂ） Methyl 5-[[2-[(2,4-dimethoxyphenyl)methylamino]-3-fluoropyridin-4-yl]methyl]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzoate (Compound a5)

Methyl 3,4-difluoro-2-(2-fluoro-4-iodoanilino)-5-[(E)-[(4-methylphenyl)sulfonylhydrazinylidene]methyl]benzoate (compound a2, 1.30 g, 2 .16 mmol), [2-[(2,4-dimethoxyphenyl)methylamino]-3-fluoropyridin-4-yl]boronic acid (compound a4, 1.98 g, 6.46 mmol) and potassium carbonate (357 mg, 2 .59 mmol) in 1,4-dioxane (59 mL) was stirred under a nitrogen atmosphere at 100° C. for 2.5 hours and then at 110° C. for 3 hours. Ethyl acetate was added to the reaction mixture and then washed with water and 13% brine. The organic layer was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and then concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (524 mg, 36%) as a foam. LCMS m/z: 682 [M+H] ⁺ . HPLC retention time: 1.03 minutes (analysis condition B)

メチル５－［［２－［（２，４－ジメトキシフェニル）メチルアミノ］－３－フルオロピリジン－４－イル］メチル］－３，４－ジフルオロ－２－（２－フルオロ－４－ヨードアニリノ）ベンゾエート（化合物ａ５、５２３ｍｇ、０．７６８ｍｍｏｌ）のＤＣＭ溶液（１６ｍＬ）を０℃に冷却し、トリフルオロ酢酸（１５．７ｍＬ）を添加し、混合物を室温で１時間撹拌した。反応混合物を減圧下で濃縮し、得られた残渣を逆相カラムクロマトグラフィー（０．０５％トリフルオロ酢酸水溶液／０．０５％トリフルオロ酢酸アセトニトリル溶液）によって精製して、表題化合物（３２１ｍｇ、７９％）をオイル状物として得た。ＬＣＭＳ ｍ／ｚ：５３２［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：０．５５分（分析条件Ｂ） Methyl 5-[(2-amino-3-fluoropyridin-4-yl)methyl]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzoate (compound a6)

Methyl 5-[[2-[(2,4-dimethoxyphenyl)methylamino]-3-fluoropyridin-4-yl]methyl]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzoate A DCM solution (16 mL) of (compound a5, 523 mg, 0.768 mmol) was cooled to 0° C., trifluoroacetic acid (15.7 mL) was added and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by reverse phase column chromatography (0.05% trifluoroacetic acid in water/0.05% trifluoroacetic acid in acetonitrile) to give the title compound (321 mg, 79 %) was obtained as an oil. LCMS m/z: 532 [M+H] ⁺ . HPLC retention time: 0.55 minutes (analysis condition B)

ＴＨＦ（６４ｍＬ）と（３２ｍＬ）中のメチル５－［（２－アミノ－３－フルオロピリジン－４－イル）メチル］－３，４－ジフルオロ－２－（２－フルオロ－４－ヨードアニリノ）ベンゾエート（化合物ａ６、４．００ｇ、７．５３ｍｍｏｌ）の混合溶液を０℃に冷却し、水酸化リチウム一水和物（９４８ｍｇ、２２．６ｍｍｏｌ）を添加し、混合物を室温で３．５時間撹拌した。０℃に冷却した後、５Ｍ塩酸（１５．１ｍＬ）を反応混合物に添加し、次いで減圧下で濃縮した。得られた残渣を水およびＴＢＭＥで洗浄して、表題化合物（４．２０ｇ、定量的）を紫色化合物として得た。ＬＣＭＳ ｍ／ｚ：５１８［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：０．６８分（分析条件Ｃ） 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (compound a7)

Methyl 5-[(2-amino-3-fluoropyridin-4-yl)methyl]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzoate in THF (64 mL) and (32 mL) A mixed solution of compound a6, 4.00 g, 7.53 mmol) was cooled to 0° C., lithium hydroxide monohydrate (948 mg, 22.6 mmol) was added and the mixture was stirred at room temperature for 3.5 hours. After cooling to 0° C., 5M hydrochloric acid (15.1 mL) was added to the reaction mixture and then concentrated under reduced pressure. The resulting residue was washed with water and TBME to give the title compound (4.20 g, quantitative) as a purple compound. LCMS m/z: 518 [M+H] ⁺ . HPLC retention time: 0.68 minutes (analysis condition C)

５－（（２－アミノ－３－フルオロピリジン－４－イル）メチル）－３，４－ジフルオロ－２－（（２－フルオロ－４－ヨードフェニル）アミノ）安息香酸塩酸塩（化合物ａ７、２００ｍｇ、０．３６１ｍｍｏｌ）の無水ＤＭＦ溶液（３．６ｍＬ）を０℃に冷却し、ＨＯＯＢｔ（６７．８ｍｇ、０．４１５ｍｍｏｌ）およびＥＤＣ・ＨＣｌ（８０．０ｍｇ、０．４１５ｍｍｏｌ）を添加し、混合物を室温で１．５時間撹拌した。ＨＯＯＢｔ（８．８ｍｇ、０．０５４ｍｍｏｌ）およびＥＤＣ・ＨＣｌ（１０．４ｍｇ、０．０５４ｍｍｏｌ）をさらに添加し、室温で１時間撹拌した後、７ＭアンモニアＭｅＯＨ溶液（０．１０３ｍＬ、０．７２２ｍｍｏｌ）およびＤＩＰＥＡ（０．１８９ｍＬ、１．０８ｍｍｏｌ）を０℃で添加し、混合物を室温で３０分間撹拌した。水と飽和炭酸水素ナトリウム水溶液を１：１で反応混合物に添加し、抽出を酢酸エチルで行った。有機層を無水硫酸ナトリウムで乾燥させ、乾燥剤を濾別した後、減圧下で濃縮した。得られた残渣を酢酸エチル（１ｍＬ）に溶解し、ヘキサン（１０ｍＬ）を添加した。得られた固体をフィルタにかけ、ヘキサンで洗浄して、表題化合物（１６２ｍｇ、８７％）を無色固体として得た。ＬＣＭＳ ｍ／ｚ：５１７［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：０．６４分（分析条件Ｃ） 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (compound a8)

5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (compound a7, 200 mg , 0.361 mmol) in anhydrous DMF (3.6 mL) was cooled to 0° C., HOOBt (67.8 mg, 0.415 mmol) and EDC.HCl (80.0 mg, 0.415 mmol) were added, and the mixture was Stir at room temperature for 1.5 hours. Additional HOOBt (8.8 mg, 0.054 mmol) and EDC.HCl (10.4 mg, 0.054 mmol) were added and stirred at room temperature for 1 hour before adding 7 M ammonia in MeOH (0.103 mL, 0.722 mmol) and DIPEA (0.189 mL, 1.08 mmol) was added at 0° C. and the mixture was stirred at room temperature for 30 minutes. Water and saturated aqueous sodium bicarbonate solution were added 1:1 to the reaction mixture and extraction was performed with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and then concentrated under reduced pressure. The resulting residue was dissolved in ethyl acetate (1 mL) and hexane (10 mL) was added. The solid obtained was filtered and washed with hexane to give the title compound (162 mg, 87%) as a colorless solid. LCMS m/z: 517 [M+H] ⁺ . HPLC retention time: 0.64 minutes (analysis condition C)

テトラキス（トリフェニルホスフィン）パラジウム（０）（１１．２ｍｇ、９．６８μｍｏｌ）および０．５Ｍシクロプロピル亜鉛ブロミド（１．９４ｍＬ、０．９６９ｍｍｏｌ）を、５－（（２－アミノ－３－フルオロピリジン－４－イル）メチル）－３，４－ジフルオロ－２－（（２－フルオロ－４－ヨードフェニル）アミノ）ベンズアミド（化合物ａ８、１００ｍｇ、０．１９４ｍｍｏｌ）の無水ＴＨＦ溶液（１．９ｍＬ）に添加し、混合物を、窒素雰囲気下、室温で２．５時間撹拌した。酢酸エチル（５ｍＬ）を反応混合物に添加し、次いでセライトでフィルタにかけ、酢酸エチル（３ｍＬ）で洗浄した。濾液を水および飽和ブラインで洗浄し、有機層を無水硫酸ナトリウムで乾燥させ、乾燥剤を濾別した後、減圧下で濃縮した。ジクロロメタン／ヘキサン（１／１０、１１ｍＬ）を得られた残渣に添加し、固体を濾別し、ヘキサン（３ｍＬ）で洗浄して、表題化合物ａ９（６３．４ｍｇ、７６％）を無色固体として得た。ＬＣＭＳ ｍ／ｚ：４３１［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：０．６１分（分析条件Ｃ） 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-cyclopropyl-2-fluorophenyl)amino)-3,4-difluorobenzamide (compound a9)

Tetrakis(triphenylphosphine)palladium(0) (11.2 mg, 9.68 μmol) and 0.5 M cyclopropylzinc bromide (1.94 mL, 0.969 mmol) were treated with 5-((2-amino-3-fluoropyridine -4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (compound a8, 100 mg, 0.194 mmol) in anhydrous THF (1.9 mL). was added and the mixture was stirred at room temperature for 2.5 hours under a nitrogen atmosphere. Ethyl acetate (5 mL) was added to the reaction mixture, which was then filtered through celite and washed with ethyl acetate (3 mL). The filtrate was washed with water and saturated brine, the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure after filtering off the drying agent. Dichloromethane/hexane (1/10, 11 mL) was added to the resulting residue and the solid was filtered off and washed with hexane (3 mL) to give title compound a9 (63.4 mg, 76%) as a colorless solid. rice field. LCMS m/z: 431 [M+H] ⁺ . HPLC retention time: 0.61 minutes (analysis condition C)

４－ニトロフェノール（５．００ｇ、３５．９ｍｍｏｌ）およびトリエチルアミン（１１．３ｍＬ、８１．０ｍｍｏｌ）のジクロロメタン溶液（６０ｍＬ）を－７８℃に冷却し、メチルスルファモイルクロリド（５．８２ｇ、４４．９ｍｍｏｌ）のジクロロメタン溶液（１５ｍＬ）を添加し、混合物を－７８℃で１．５時間撹拌した。反応混合物を減圧下で濃縮し、得られた残渣をシリカゲル・カラム・クロマトグラフィー（ヘキサン／酢酸エチル）および逆相カラムクロマトグラフィー（０．１％ギ酸水溶液／０．１％ギ酸アセトニトリル溶液）によって精製して、表題化合物（５．５１ｇ、６６％）を無色固体として得た。ＨＰＬＣ保持時間：０．６３分（分析条件Ｃ）^１Ｈ－ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：８．３１（２Ｈ，ｍ），７．４６（２Ｈ，ｍ），４．６８（１Ｈ，ｍ），３．００（３Ｈ，ｄ，Ｊ＝５．４Ｈｚ）． 4-nitrophenyl methylsulfamate (compound r1)

A dichloromethane solution (60 mL) of 4-nitrophenol (5.00 g, 35.9 mmol) and triethylamine (11.3 mL, 81.0 mmol) was cooled to −78° C. and methylsulfamoyl chloride (5.82 g, 44.0 mmol) was added. 9 mmol) in dichloromethane (15 mL) was added and the mixture was stirred at −78° C. for 1.5 hours. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane/ethyl acetate) and reverse-phase column chromatography (0.1% formic acid aqueous solution/0.1% formic acid acetonitrile solution). to give the title compound (5.51 g, 66%) as a colorless solid. HPLC retention time: 0.63 minutes (analysis condition C) ¹ H-NMR (400 MHz, CDCl ₃ ) δ: 8.31 (2H, m), 7.46 (2H, m), 4.68 (1H, m ), 3.00 (3H, d, J=5.4 Hz).

５－（（２－アミノ－３－フルオロピリジン－４－イル）メチル）－２－（（４－シクロプロピル－２－フルオロフェニル）アミノ）－３，４－ジフルオロベンズアミド（化合物ａ９、２．４７ｇ、５．７４ｍｍｏｌ）を無水ＤＭＦ（２８．７ｍＬ）に溶解した後、ピリジン（２．７８ｍＬ、３４．４ｍｍｏｌ）および４－ニトロフェニルメチルスルファメート（化合物ｒ１、４．００ｇ、１７．２ｍｍｏｌ）を添加し、混合物を４０℃で２．５時間撹拌した。反応混合物を室温に冷却し、水（２４．７ｍＬ）を添加した。アセトニトリル（３ｍＬ）および水（１９．８ｍＬ）をさらに添加し、１０分間撹拌した後、固体を濾別した。得られた固体を水／アセトニトリル（１／１、４９．４ｍＬ）で洗浄して、表題化合物（２．５６ｇ、８５％）を無色固体として得た。ＬＣＭＳ ｍ／ｚ：５２４［Ｍ＋Ｈ］^＋．ＨＰＬＣ保持時間：１．１３分（分析条件Ａ） 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl]benzamide (formula ( II) compound)

5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-cyclopropyl-2-fluorophenyl)amino)-3,4-difluorobenzamide (compound a9, 2.47 g , 5.74 mmol) was dissolved in anhydrous DMF (28.7 mL) followed by pyridine (2.78 mL, 34.4 mmol) and 4-nitrophenylmethylsulfamate (compound r1, 4.00 g, 17.2 mmol). was added and the mixture was stirred at 40° C. for 2.5 hours. The reaction mixture was cooled to room temperature and water (24.7 mL) was added. Additional acetonitrile (3 mL) and water (19.8 mL) were added and after stirring for 10 minutes the solids were filtered off. The solid obtained was washed with water/acetonitrile (1/1, 49.4 mL) to give the title compound (2.56 g, 85%) as a colorless solid. LCMS m/z: 524 [M+H] ⁺ . HPLC retention time: 1.13 minutes (analysis condition A)

2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl]benzamide sodium salt ( Compound IIa)

(1) Preparation of Compound IIa (Form I) Acetone (10.6 mL) and DMSO (1.51 mL) were combined with 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[ 3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl]benzamide ( compound of formula (II) , 3.03 g) was added and dissolved at room temperature. A 20% sodium ethoxide solution in ethanol (3.03 mL) and seed crystals of the sodium salt of Compound II (Sample IIb described below) were added to the solution and the mixture was stirred at room temperature for 1 hour, after which ethanol (15.1 mL) was added. was added and the mixture was stirred at room temperature for 4 hours. Ethanol (15.1 mL) was then added and the mixture was stirred at room temperature for 4 hours to give compound II sodium salt (2.74 g) as powdery crystals (Sample IIa (Form I)).

(2) Preparation of sample IIb 20% sodium ethoxide ethanol solution (0.054 mL) and methyl isobutyl ketone (0.161 mL) were added to the compound of formula (II) (53.6 mg) and the mixture was stirred at room temperature for 30 minutes. Stir, then add methyl isobutyl ketone (0.161 mL) and continue stirring at 60° C. for 4 days. DMSO (0.054 mL) was then added and the mixture was stirred at 60° C. for 5 hours to give compound II sodium salt (25.6 mg) as powdery crystals (Sample IIb).

MEK1 Inhibitory Activity of Compound II The MEK1 inhibitory activity of Compound II was evaluated by fluorescence polarization as follows.

Test compounds, CRAF (Thermo Fisher Scientific Inc.), MEK1 (Thermo Fisher Scientific Inc.) and ERK2 (Carna Biosciences, Inc.) were mixed in ATP-containing buffer and allowed to react at 30° C. for 60 minutes. FAM-labeled ERKtide (Molecular Devices Corp.) was then added and the reaction continued at 30° C. for 45 minutes. Additional IMAP® Progressive Binding Reagent (Molecular Devices Corp.) was added and the reaction continued at room temperature for 15 minutes. After the reaction, fluorescence polarization was measured using a fluorescence plate reader and the 50% inhibitory concentration (IC50) was calculated based on percent inhibition relative to controls without test compound. When the test compound was Compound II, an IC50 of 17 nM was measured for MEK1 activity.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である、ＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
式（Ｉ）の化合物が、（３Ｒ）－Ｎ－［２－シアノ－４－フルオロ－３－（３－メチル－４－オキソ－キナゾリン－６－イル）オキシ－フェニル］－３－フルオロ－ピロリジン－１－スルホンアミドである、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＭＥＫ阻害剤が、２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミド、コビメチニブ、トラメチニブおよびビニメチニブ、またはそれらの薬学的に許容され得る塩から選択される、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＭＥＫ阻害剤が、２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミド、またはその薬学的に許容され得る塩である、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＭＥＫ阻害剤が２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミドナトリウム塩である、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＭＥＫ阻害剤が、コビメチニブまたはその薬学的に許容され得る塩である、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＭＥＫ阻害剤が［３，４－ジフルオロ－２－（２－フルオロ－４－ヨードアニリノ）フェニル］－［３－ヒドロキシ－３－［（２Ｓ）－ピペリジン－２－イル］アゼチジン－１－イル］メタノンヘミフマル酸塩である、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＭＥＫ阻害剤が［３，４－ジフルオロ－２－（２－フルオロ－４－ヨードアニリノ）フェニル］－［３－ヒドロキシ－３－［（２Ｓ）－ピペリジン－２－イル］アゼチジン－１－イル］メタノンヘミコハク酸塩である、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
医薬として使用するための、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
がんの治療的および／または予防的処置に使用するための、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
がんの治療または予防のための医薬を調製するための、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せの使用；
がん、特に黒色腫または非小細胞肺がんの治療または予防のための方法であって、有効量の、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せを、それを必要とする患者に投与することを含む、方法；
本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ、および１種以上の薬学的に許容され得る添加物を含む医薬組成物；
ＢＲＡＦ阻害剤およびＭＥＫ阻害剤が両方とも経口投与される、本明細書に記載の組合せ、使用、方法または医薬組成物；
ＢＲＡＦ阻害剤がＭＥＫ阻害剤と同時に投与される、本明細書に記載の組合せ、使用、方法または医薬組成物；
ＢＲＡＦ阻害剤およびＭＥＫ阻害剤が共製剤化されている、本発明に記載の組合せ、使用、方法または医薬組成物；
ＢＲＡＦ阻害剤がＭＥＫ阻害剤と逐次的に投与される、本発明に記載の組合せ、使用、方法または医薬組成物；
がんが、甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんである、本発明に記載の使用のためのＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
がんがＢＲＡＦ^Ｖ６００変異に関連する、本発明に記載の使用のための、ＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
がんがＢＲＡＦ^Ｖ６００変異陽性の切除不能なまたは転移性のがんである、本発明に記載の使用のための、ＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＢＲＡＦ^Ｖ６００変異が、（ａ）患者の腫瘍組織および／または体液の試料から抽出された核酸（例えばＤＮＡ）に対してＰＣＲまたは配列決定を行うこと；および（ｂ）試料中のＢＲＡＦ^Ｖ６００の発現を決定することを含む方法を使用して決定される、本明細書に記載の使用のための、ＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＭＥＫ分解剤、ＥＧＦＲ阻害剤、ＥＧＦＲ分解剤、ＨＥＲ２および／またはＨＥＲ３の阻害剤、ＨＥＲ２および／またはＨＥＲ３の分解剤、ＳＨＰ２阻害剤、ＳＨＰ２分解剤、Ａｘｌ阻害剤、Ａｘｌ分解剤、ＡＬＫ阻害剤、ＡＬＫ分解剤、ＰＩ３Ｋ阻害剤、ＰＩ３Ｋ分解剤、ＳＯＳ１阻害剤、ＳＯＳ１分解剤、シグナル伝達経路阻害剤、チェックポイント阻害剤、アポトーシス経路の調節剤、細胞傷害性化学療法剤、血管新生標的治療薬、免疫標的剤、ならびに抗体－薬物コンジュゲートから選択される１種以上のさらなる抗がん剤を含む、本発明に記載の使用のための、ＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＢＲＡＦ阻害剤が、（３Ｒ）－Ｎ－［２－シアノ－４－フルオロ－３－（３－メチル－４－オキソ－キナゾリン－６－イル）オキシ－フェニル］－３－フルオロ－ピロリジン－１－スルホンアミドまたはその薬学的に許容され得る塩である、がんの治療または予防のための医薬を調製するための、本明細書に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せの使用；
ＭＥＫ阻害剤が、２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミド、コビメチニブ、トラメチニブおよびビニメチニブから、またはその薬学的に許容され得る塩から選択される、本発明に記載の医薬を調製するための、本発明に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤との組合せ；
ＢＲＡＦ阻害剤が、（３Ｒ）－Ｎ－［２－シアノ－４－フルオロ－３－（３－メチル－４－オキソ－キナゾリン－６－イル）オキシ－フェニル］－３－フルオロ－ピロリジン－１－スルホンアミドまたはその薬学的に許容され得る塩である、がんの治療または予防のための方法であって、有効量の、本明細書に記載のＢＲＡＦ阻害剤とＭＥＫ阻害剤とを、それを必要とする患者に投与することを含む、方法；
がんが、甲状腺がん、結腸直腸がん、黒色腫、脳のがんおよび非小細胞肺がんから選択される、本発明に記載のがんの治療または予防のための方法；
ＭＥＫ阻害剤が、２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミド、コビメチニブ、トラメチニブおよびビニメチニブから、またはその薬学的に許容され得る塩から選択される、本発明に記載のがんの治療または予防のための方法；
ＭＥＫ阻害剤が、２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミド、コビメチニブ、ビニメチニブ、トラメチニブ、セルメチニブ、ピマセルチブ、レファメチニブ、Ｎ－［２（Ｒ），３－ジヒドロキシプロポキシ］－３，４－ジフルオロ－２－（２－フルオロ－４－ヨードフェニルアミノ）ベンズアミド（ＰＤ－３２５９０１）、２－（２－クロロ－４－ヨードフェニルアミノ）－Ｎ－（シクロプロピルメトキシ）－３，４－ジフルオロベンズアミド（Ｃｌ－１０４０）および３－［２（Ｒ），３－ジヒドロキシプロピル］－６－フルオロ－５－（２－フルオロ－４－ヨードフェニルアミノ）－８－メチルピリド［２，３－ｄ］ピリミジン－４，７（３Ｈ，８Ｈ）－ジオン（ＴＡＫ－７３３）から選択される、本明細書に記載の医薬組成物；
ＭＥＫ阻害剤が、２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミド、コビメチニブ、トラメチニブおよびビニメチニブから、またはその薬学的に許容され得る塩から選択される、本明細書に記載の医薬組成物；
ＭＥＫ阻害剤が、２－（４－シクロプロピル－２－フルオロアニリノ）－３，４－ジフルオロ－５－［［３－フルオロ－２－（メチルスルファモイルアミノ）ピリジン－４－イル］メチル］ベンズアミドまたはその薬学的に許容され得る塩である、本明細書に記載の医薬組成物；
ＭＥＫ阻害剤が、コビメチニブまたはその薬学的に許容され得る塩である、本明細書に記載の医薬組成物；
式（Ｉ）の化合物が（３Ｒ）－Ｎ－［２－シアノ－４－フルオロ－３－（３－メチル－４－オキソ－キナゾリン－６－イル）オキシ－フェニル］－３－フルオロ－ピロリジン－１－スルホンアミドである、本明細書に記載の医薬組成物；
式（Ｉ）の化合物が（３Ｓ）－Ｎ－［２－シアノ－４－フルオロ－３－（３－メチル－４－オキソ－キナゾリン－６－イル）オキシ－フェニル］－３－フルオロ－ピロリジン－１－スルホンアミドである、本明細書に記載の医薬組成物；
がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
ＢＲＡＦ阻害剤およびＭＥＫ阻害剤が経口投与される、がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
第１の組成物が第２の組成物と同時に投与される、がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
第１および第２の組成物が共製剤化されている、がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
第１の組成物が第２の組成物と逐次的に投与される、がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
がんがＢＲＡＦ変異に関連する、がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
がんが、ＢＲＡＦ^Ｖ６００変異陽性の切除不能または転移性のがん、特にＢＲＡＦ^{Ｖ６００Ｅ}またはＢＲＡＦ^{Ｖ６００Ｋ}変異陽性の切除不能なまたは転移性のがんである、がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
ＢＲＡＦ^Ｖ６００変異が、（ａ）患者の腫瘍組織および／または体液の試料から抽出された核酸（例えばＤＮＡ）に対してＰＣＲまたは配列決定を行うこと；および（ｂ）試料中のＢＲＡＦ^Ｖ６００の発現を決定することを含む方法を使用して決定される、がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防に使用するための、本明細書に記載の医薬組成物；
がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防における本明細書に記載の医薬組成物の使用；ならびに
がん、特に甲状腺がん、結腸直腸がん、黒色腫、脳のがんまたは非小細胞肺がんの治療または予防のための医薬を調製するための、本明細書に記載の医薬組成物の使用。 The invention particularly relates to:
The BRAF inhibitor is of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, a combination of a BRAF inhibitor and a MEK inhibitor;
The compound of formula (I) is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine - a combination of a BRAF inhibitor and a MEK inhibitor according to the present invention, which is a 1-sulfonamide;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl ] a combination of a BRAF inhibitor and a MEK inhibitor according to the present invention selected from benzamide, cobimetinib, trametinib and binimetinib, or pharmaceutically acceptable salts thereof;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl ] a combination of a BRAF inhibitor and a MEK inhibitor according to the present invention, which is a benzamide, or a pharmaceutically acceptable salt thereof;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl] a combination of a BRAF inhibitor and a MEK inhibitor according to the invention, which is a benzamide sodium salt;
A combination of a BRAF inhibitor and a MEK inhibitor according to the present invention, wherein the MEK inhibitor is cobimetinib or a pharmaceutically acceptable salt thereof;
The MEK inhibitor is [3,4-difluoro-2-(2-fluoro-4-iodoanilino)phenyl]-[3-hydroxy-3-[(2S)-piperidin-2-yl]azetidin-1-yl]meth a combination of a BRAF inhibitor and a MEK inhibitor according to the present invention, which is non-hemifumarate;
The MEK inhibitor is [3,4-difluoro-2-(2-fluoro-4-iodoanilino)phenyl]-[3-hydroxy-3-[(2S)-piperidin-2-yl]azetidin-1-yl]meth a combination of a BRAF inhibitor and a MEK inhibitor according to the present invention, which is non-hemisuccinate;
a combination of a BRAF inhibitor and a MEK inhibitor according to the invention for use as a medicament;
a combination of a BRAF inhibitor and a MEK inhibitor according to the invention for use in the therapeutic and/or prophylactic treatment of cancer;
use of a combination of a BRAF inhibitor and a MEK inhibitor according to the invention for the preparation of a medicament for the treatment or prevention of cancer;
A patient in need of a method for the treatment or prevention of cancer, particularly melanoma or non-small cell lung cancer, in an effective amount of a combination of a BRAF inhibitor and a MEK inhibitor according to the present invention. a method comprising administering to;
A pharmaceutical composition comprising a combination of a BRAF inhibitor and a MEK inhibitor according to the invention and one or more pharmaceutically acceptable excipients;
a combination, use, method or pharmaceutical composition described herein wherein the BRAF inhibitor and the MEK inhibitor are both orally administered;
a combination, use, method or pharmaceutical composition described herein, wherein the BRAF inhibitor is administered concurrently with the MEK inhibitor;
a combination, use, method or pharmaceutical composition according to the invention, wherein a BRAF inhibitor and a MEK inhibitor are co-formulated;
a combination, use, method or pharmaceutical composition according to the invention, wherein the BRAF inhibitor is administered sequentially with the MEK inhibitor;
a combination of a BRAF inhibitor and a MEK inhibitor for use according to the invention wherein the cancer is thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer;
a combination of a BRAF inhibitor and a MEK inhibitor for use according to the invention wherein the cancer is associated with a BRAF ^V600 mutation;
a combination of a BRAF inhibitor and a MEK inhibitor for use according to the invention, wherein the cancer is BRAF ^V600 mutation-positive unresectable or metastatic cancer;
BRAF ^V600 mutations are detected by (a) PCR or sequencing nucleic acid (e.g., DNA) extracted from a sample of patient tumor tissue and/or body fluid; and (b) expression of BRAF ^V600 in the sample. a combination of a BRAF inhibitor and a MEK inhibitor for use as described herein as determined using a method comprising determining;
MEK degrading agents, EGFR inhibitors, EGFR degrading agents, HER2 and/or HER3 inhibitors, HER2 and/or HER3 degrading agents, SHP2 inhibitors, SHP2 degrading agents, Axl inhibitors, Axl degrading agents, ALK inhibitors, ALK degrading agents, PI3K inhibitors, PI3K degrading agents, SOS1 inhibitors, SOS1 degrading agents, signal transduction pathway inhibitors, checkpoint inhibitors, regulators of apoptotic pathways, cytotoxic chemotherapeutic agents, angiogenesis targeting therapeutic agents, a combination of a BRAF inhibitor and a MEK inhibitor for use according to the present invention comprising one or more additional anti-cancer agents selected from immunotargeting agents and antibody-drug conjugates;
The BRAF inhibitor is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1- use of a combination of a BRAF inhibitor and a MEK inhibitor as described herein for the preparation of a medicament for treating or preventing cancer, which is a sulfonamide or a pharmaceutically acceptable salt thereof;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl ] a BRAF inhibitor according to the invention and a MEK inhibitor for the preparation of a medicament according to the invention, selected from benzamide, cobimetinib, trametinib and binimetinib, or from pharmaceutically acceptable salts thereof. combination;
The BRAF inhibitor is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1- A method for the treatment or prevention of cancer, which is a sulfonamide or a pharmaceutically acceptable salt thereof, comprising an effective amount of a BRAF inhibitor and a MEK inhibitor as described herein, a method comprising administering to a patient in need thereof;
A method for the treatment or prevention of cancer according to the invention, wherein the cancer is selected from thyroid cancer, colorectal cancer, melanoma, brain cancer and non-small cell lung cancer;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl ] methods for the treatment or prevention of cancer according to the present invention selected from benzamide, cobimetinib, trametinib and binimetinib, or from pharmaceutically acceptable salts thereof;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl ] benzamide, cobimetinib, binimetinib, trametinib, selumetinib, pimasertib, lefametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide ( PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (Cl-1040) and 3-[2(R),3-dihydroxy selected from propyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733) a pharmaceutical composition as described herein;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl ] a pharmaceutical composition as described herein, selected from benzamide, cobimetinib, trametinib and binimetinib, or from a pharmaceutically acceptable salt thereof;
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl ] the pharmaceutical composition described herein, which is a benzamide or a pharmaceutically acceptable salt thereof;
the pharmaceutical composition described herein, wherein the MEK inhibitor is cobimetinib or a pharmaceutically acceptable salt thereof;
The compound of formula (I) is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine- A pharmaceutical composition as described herein which is a 1-sulfonamide;
The compound of formula (I) is (3S)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine- A pharmaceutical composition as described herein which is a 1-sulfonamide;
a pharmaceutical composition as described herein for use in the treatment or prevention of cancer, in particular thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer;
BRAF inhibitors and MEK inhibitors are orally administered herein for use in the treatment or prevention of cancer, particularly thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer. a pharmaceutical composition as described in
For use in the treatment or prevention of cancer, particularly thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer, wherein a first composition is administered concurrently with a second composition of the pharmaceutical composition described herein;
for use in the treatment or prevention of cancer, particularly thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer, wherein the first and second compositions are co-formulated , a pharmaceutical composition as described herein;
Use in the treatment or prevention of cancer, particularly thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer, wherein a first composition is administered sequentially with a second composition A pharmaceutical composition as described herein for
a pharmaceutical composition as described herein for use in the treatment or prevention of cancer, in particular thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer;
A pharmaceutical composition as described herein for use in treating or preventing cancer, wherein the cancer is associated with BRAF mutations;
for use in the treatment or prevention of cancer, wherein the cancer is a BRAF ^V600 mutation-positive unresectable or metastatic cancer, in particular a BRAF ^V600E or BRAF ^V600K mutation-positive unresectable or metastatic cancer; a pharmaceutical composition as described herein;
BRAF ^V600 mutations are detected by (a) PCR or sequencing nucleic acid (e.g., DNA) extracted from a sample of patient tumor tissue and/or body fluid; and (b) expression of BRAF ^V600 in the sample. for use in the treatment or prevention of cancer, particularly thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer, determined using a method comprising determining a pharmaceutical composition as described herein;
Use of the pharmaceutical compositions described herein in the treatment or prevention of cancer, especially thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer; and cancer, especially thyroid cancer. , colorectal cancer, melanoma, brain cancer or non-small cell lung cancer.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である、化合物。 Further embodiments of the invention are as follows:
[1] A compound for use in treating or preventing cancer in combination with a MEK inhibitor, comprising formula (I):

or a pharmaceutically acceptable salt or solvate thereof.

[2] A pharmaceutical composition comprising the compound of [1] and one or more pharmaceutically acceptable additives for treating or preventing cancer in combination with a MEK inhibitor.

[3] Use of the compound of [1] in the manufacture of a medicament for treating or preventing cancer in combination with a MEK inhibitor.

[4] A method for treating or preventing cancer, comprising administering an effective amount of a combination of a MEK inhibitor and the compound of [1] to a patient in need thereof, Method.

[5] The compound of formula (I) is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro -pyrrolidine-1-sulfonamide, the compound, pharmaceutical composition, use or method of any one of [1] to [4].

[6] the MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine-4- yl]methyl]benzamide, cobimetinib, trametinib and binimetinib, or a pharmaceutically acceptable salt or solvate thereof, the compound, pharmaceutical composition according to any one of [1] to [5] , use or method.

[7] the MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine-4- The compound, pharmaceutical composition, use or method of any one of [1] to [6], which is yl]methyl]benzamide or a pharmaceutically acceptable salt or solvate thereof.

[7.1] The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine- The compound, pharmaceutical composition, use or method of any one of [1] to [7], which is 4-yl]methyl]benzamide or a pharmaceutically acceptable salt thereof.

[7.2] The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine-4 -yl]methyl]benzamide sodium salt, the compound, pharmaceutical composition, use or method of any one of [1] to [7.1].

[8] The compound, pharmaceutical composition, use or method of any one of [1] to [6], wherein the MEK inhibitor is cobimetinib or a pharmaceutically acceptable salt or solvate thereof.

[9] The compound, pharmaceutical composition, use or method of any one of [1] to [8], wherein both the compound of formula (I) and the MEK inhibitor are orally administered.

[10] The compound, pharmaceutical composition, use or method of any one of [1] to [9], wherein the compound of formula (I) is administered concurrently with a MEK inhibitor.

[11] The compound, pharmaceutical composition, use or method of any one of [1] to [10], wherein the compound of formula (I) is administered sequentially with the MEK inhibitor.

[12] A compound for use in treating or preventing cancer in combination with a BRAF inhibitor, comprising: 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[ A compound that is 3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl]benzamide or a pharmaceutically acceptable salt or solvate thereof.

[13] A pharmaceutical composition comprising the compound of [12] and one or more pharmaceutically acceptable additives for treating or preventing cancer in combination with a BRAF inhibitor.

[14] Use of the compound of [12] in the manufacture of a medicament for use in treating or preventing cancer in combination with a BRAF inhibitor.

[15] A method for treating or preventing cancer, comprising administering an effective amount of a combination of a BRAF inhibitor and the compound of [12] to a patient in need thereof, Method.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である、［１２］から［１５］のいずれか１つに記載の化合物、医薬組成物、使用または方法。 [16] The BRAF inhibitor has the formula (I):

or a pharmaceutically acceptable salt or solvate thereof, the compound, pharmaceutical composition, use or method of any one of [12] to [15].

[17] The compound of formula (I) is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro -pyrrolidine-1-sulfonamide, the compound, pharmaceutical composition, use or method of [16].

[18] The compound, pharmaceutical composition, use or method of any one of [12] to [17], wherein both the compound of [12] and the BRAF inhibitor are orally administered.

[19] The compound, pharmaceutical composition, use or method of any one of [12] to [18], wherein the compound of [12] is administered concurrently with a BRAF inhibitor.

[20] The compound, pharmaceutical composition, use or method of any one of [12] to [19], wherein the compound of [12] is administered sequentially with a BRAF inhibitor.

[21] The compound according to [12] is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino) The compound, pharmaceutical composition, use or method of any one of [12] to [20], which is pyridin-4-yl]methyl]benzamide or a pharmaceutically acceptable salt thereof.

[22] The compound of [12] is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine -4-yl]methyl]benzamide sodium salt, the compound, pharmaceutical composition, use or method of any one of [12] to [21].

[23] The compound or pharmaceutical composition of any one of [1] to [22], wherein the cancer is thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer an object, use or method.

[24] The compound, pharmaceutical composition, use or method of any one of [1] to [23], wherein the cancer is associated with BRAFV600 mutations.

[25] The compound, pharmaceutical composition, use or method of any one of [1] to [24], wherein the cancer is BRAFV600 mutation-positive unresectable or metastatic cancer.

[26] The BRAFV600 mutation is (a) PCR or sequenced on nucleic acid (e.g., DNA) extracted from a sample of patient tumor tissue and/or body fluid; and (b) expression of BRAFV600 in the sample. The compound, pharmaceutical composition, use or method of any one of [1] to [25], wherein the compound, pharmaceutical composition, use or method of any one of [1] to [25] is determined using a method comprising determining the

[27] MEK degrading agent, EGFR inhibitor, EGFR degrading agent, HER2 and/or HER3 inhibitor, HER2 and/or HER3 degrading agent, SHP2 inhibitor, SHP2 degrading agent, Axl inhibitor, Axl degrading agent, ALK inhibitors, ALK degrading agents, PI3K inhibitors, PI3K degrading agents, SOS1 inhibitors, SOS1 degrading agents, signal transduction pathway inhibitors, checkpoint inhibitors, modulators of apoptotic pathways, cytotoxic chemotherapeutic agents, angiogenic targets a compound, pharmaceutical composition, according to any one of [1] to [26], comprising one or more additional anticancer agents selected from therapeutic agents, immunotargeting agents, and antibody-drug conjugates; use or method.

の化合物またはその薬学的に許容され得る塩もしくは溶媒和物である、組合せ。 [101] A combination of a BRAF inhibitor and a MEK inhibitor, wherein the MEK inhibitor is

or a pharmaceutically acceptable salt or solvate thereof.

[102] The compound of formula (II) is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine A combination of a BRAF inhibitor and a MEK inhibitor according to [101], which is 4-yl]methyl]benzamide.

[103] The BRAF inhibitor is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine - A combination of a BRAF inhibitor and a MEK inhibitor according to [101] or [102], selected from 1-sulfonamide, vemurafenib, dabrafenib, and encorafenib, or a pharmaceutically acceptable salt thereof.

[104] The BRAF inhibitor is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine - A combination of a BRAF inhibitor and a MEK inhibitor according to any one of [101] to [103], which is a 1-sulfonamide or a pharmaceutically acceptable salt thereof.

[105] The combination of a BRAF inhibitor and a MEK inhibitor of any one of [101] to [103], wherein the BRAF inhibitor is encorafenib or a pharmaceutically acceptable salt thereof.

[106] A combination of the BRAF inhibitor of any one of [101] to [105] and a MEK inhibitor for use as a medicament.

[107] A combination of a BRAF inhibitor and a MEK inhibitor according to any one of [101] to [105] for use in therapeutic and/or prophylactic treatment of cancer.

[108] Use of the combination of the BRAF inhibitor and MEK inhibitor of any one of [101] to [105] for preparing a medicament for treating or preventing cancer.

[109] A method for the treatment or prevention of cancer, particularly melanoma or non-small cell lung cancer, comprising an effective amount of the BRAF inhibitor of any one of [101] to [105] and MEK A method comprising administering a combination with an inhibitor to a patient in need thereof.

[110] A pharmaceutical composition comprising the combination of the BRAF inhibitor and MEK inhibitor of any one of [101] to [105], and one or more pharmaceutically acceptable additives.

[111] The combination, use, method or pharmaceutical composition of any one of [106] to [110], wherein said BRAF inhibitor and said MEK inhibitor are both orally administered.

[112] The combination, use, method or pharmaceutical composition of any one of [106] to [111], wherein said BRAF inhibitor is administered concurrently with said MEK inhibitor.

[113] The combination, use, method or pharmaceutical composition of any one of [106] to [112], wherein said BRAF inhibitor and said MEK inhibitor are co-formulated.

[114] The combination, use, method or pharmaceutical composition of any one of [106] to [111], wherein said BRAF inhibitor is administered sequentially with said MEK inhibitor.

[115] For the use of any one of [106] to [108], wherein said cancer is thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer of a BRAF inhibitor and a MEK inhibitor.

[116] A combination of a BRAF inhibitor and a MEK inhibitor for use according to any one of [106] to [109], wherein said cancer is associated with BRAF ^V600 mutations.

[117] A BRAF inhibitor and a MEK inhibitor for use according to any one of [106] to [109], wherein said cancer is BRAF ^V600 mutation-positive unresectable or metastatic cancer combination with.

[118] The BRAF ^V600 mutation is determined by (a) PCR or sequencing nucleic acid (e.g., DNA) extracted from a sample of patient tumor tissue and/or body fluid; and (b) BRAF in said sample. A combination of a BRAF inhibitor and a MEK inhibitor for use according to any one of [106] to [109], determined using a method comprising determining expression of ^V600 .

[119] MEK degrading agents, EGFR inhibitors, EGFR degrading agents, inhibitors of HER2 and/or HER3, degrading agents of HER2 and/or HER3, SHP2 inhibitors, SHP2 degrading agents, Axl inhibitors, Axl degrading agents, ALK inhibitors, ALK degrading agents, PI3K inhibitors, PI3K degrading agents, SOS1 inhibitors, SOS1 degrading agents, signal transduction pathway inhibitors, checkpoint inhibitors, modulators of apoptotic pathways, cytotoxic chemotherapeutic agents, angiogenic targets BRAF inhibition for use according to any one of [106] to [109], comprising one or more additional anticancer agents selected from therapeutic agents, immunotargeting agents, and antibody-drug conjugates A combination of a drug and a MEK inhibitor.

[120] the MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine-4 -yl]methyl]benzamide or a pharmaceutically acceptable salt thereof.

[121] the MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine-4 The combination, use, method or pharmaceutical composition of any one of [101] to [120], which is -yl]methyl]benzamide.

[122] the MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridine-4 The combination, use, method or pharmaceutical composition of any one of [101] to [120], which is -yl]methyl]benzamide sodium salt.

A particular embodiment of the invention is a method for the treatment or prevention of cancer, particularly thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer, comprising an effective amount of , to a method comprising administering a pharmaceutical composition as described herein to a patient in need thereof;
Certain embodiments of the present invention relate to the pharmaceutical composition described herein for use as a medicament for the treatment and/or prevention of brain metastases;
A particular embodiment of the present invention relates to a medicament as described herein, for use as a medicament for the treatment and/or prevention of brain metastasis, wherein the primary tumor is melanoma or non-small cell lung cancer. relating to the composition;
Certain embodiments of the present invention are directed to a medicament as described herein for use in the treatment and/or prevention of cancer, particularly melanoma or non-small cell lung cancer, in which the patient is naive to targeted therapy. relating to the composition;
Certain embodiments of the present invention are used in the treatment and/or prevention of cancer, particularly melanoma or non-small cell lung cancer, in which the patient is treatment naive with respect to targeted therapy and checkpoint inhibitor treatment naive. Relating to the pharmaceutical compositions described herein for
Certain embodiments of the present invention are used in the treatment and/or prevention of cancer, particularly melanoma or non-small cell lung cancer, where the patient is treatment-experienced with targeted therapy and treatment-experienced with checkpoint inhibitors. Relating to the pharmaceutical compositions described herein for
Certain embodiments of the present invention are directed to the therapeutic agents described herein for use in the treatment and/or prevention of cancer, particularly melanoma or non-small cell lung cancer, wherein the cancer has been previously treated by surgery. for the pharmaceutical composition of
Certain embodiments of the invention are described herein for use in the treatment and/or prevention of cancer, particularly melanoma or non-small cell lung cancer, in which the patient is naive to BRAF inhibitor therapy. for the pharmaceutical composition of
Certain embodiments of the present invention relate to a pharmaceutical composition as described herein for use as a medicament for the treatment and/or treatment of BRAF inhibitor-resistant tumors;
Certain embodiments of the present invention are described herein for use in the treatment and/or prevention of cancer, particularly melanoma or non-small cell lung cancer, in which the patient is naive to MEK inhibitor treatment. and certain embodiments of the present invention relate to pharmaceutical compositions described herein for use as a medicament for the treatment and/or prevention of MEK inhibitor-resistant tumors .

Certain embodiments of the invention provide treatment of cancer in subjects previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib and encorafenib and/or a MEK inhibitor selected from binimetinib, trametinib and cobimetinib and/or to a pharmaceutical composition as described herein for use as a medicament for prophylaxis;
Certain embodiments of the present invention relate to pharmaceutical compositions described herein for use as a medicament for treatment and/or prevention in subjects previously treated with encorafenib and binimetinib;
Certain embodiments of the present invention relate to pharmaceutical compositions described herein for use as a medicament for treatment and/or prevention in subjects previously treated with dabrafenib and trametinib;
Certain embodiments of the present invention relate to pharmaceutical compositions described herein for use as a medicament for treatment and/or prevention in subjects previously treated with vemurafenib and cobimetinib;
Certain embodiments of the present invention provide the present invention for use as a medicament for the treatment and/or prevention of cancer, particularly melanoma or non-small cell lung cancer, in subjects previously treated with checkpoint inhibitors. relating to the pharmaceutical compositions described herein;
Certain embodiments of the present invention are described herein for use in the treatment and/or prevention of cancer cancer, particularly melanoma or non-small cell lung cancer, in which the patient is naive to targeted therapy. with respect to the BRAF inhibitors and MEK inhibitors described;
Certain embodiments of the present invention provide therapeutic and/or prophylactic treatment of cancer, particularly melanoma or non-small cell lung cancer, in which the patient is treatment naïve with respect to targeted therapy and checkpoint inhibitor naïve. BRAF inhibitors and MEK inhibitors as described herein for use in
Certain embodiments of the present invention provide for use in the therapeutic and/or prophylactic treatment of cancer, wherein the patient is treatment-experienced with targeted therapy and checkpoint inhibitor treatment-experienced herein. BRAF inhibitors and MEK inhibitors according to;
Certain embodiments of the present invention are directed to therapeutic agents herein for use in the therapeutic and/or prophylactic treatment of cancer, particularly melanoma or non-small cell lung cancer, in which the patient is naïve to BRAF inhibitor therapy. BRAF inhibitors and MEK inhibitors described in
Certain embodiments of the present invention relate to BRAF inhibitors and MEK inhibitors as described herein for use as medicaments for the treatment and/or prevention of BRAF inhibitor-resistant tumors;
Certain embodiments of the present invention are directed to therapeutic agents herein for use in the therapeutic and/or prophylactic treatment of cancer, particularly melanoma or non-small cell lung cancer, in which the patient is naïve to MEK inhibitor treatment. BRAF inhibitors and MEK inhibitors described in
Certain embodiments of the present invention relate to BRAF inhibitors and MEK inhibitors as described herein for use in therapeutic and/or prophylactic treatment of MEK inhibitor-resistant tumors;
Certain embodiments of the invention provide treatment of cancer in subjects previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib and encorafenib and/or a MEK inhibitor selected from binimetinib, trametinib and cobimetinib and/or BRAF inhibitors and MEK inhibitors as described herein for use as a medicament for prophylaxis;
Certain embodiments of the present invention relate to BRAF inhibitors and MEK inhibitors as described herein for use as medicaments for treatment and/or prevention in subjects previously treated with encorafenib and binimetinib ;
Certain embodiments of the present invention relate to BRAF inhibitors and MEK inhibitors as described herein for use as medicaments for treatment and/or prophylaxis in subjects previously treated with dabrafenib and trametinib ;
Certain embodiments of the present invention relate to BRAF inhibitors and MEK inhibitors as described herein for use as medicaments for treatment and/or prevention in subjects previously treated with vemurafenib and cobimetinib ;
Certain embodiments of the present invention are directed to compounds described herein for use in therapeutic and/or prophylactic treatment of cancer, particularly melanoma or non-small cell lung cancer, in subjects previously treated with checkpoint inhibitors. BRAF inhibitors and MEK inhibitors described in
In one embodiment, the invention provides a kit comprising a BRAF inhibitor and a MEK inhibitor as described herein, prescribing information, also known as a "leaflet", a blister package or bottle (HDPE or glass) and a container. . The prescribing information preferably includes advice to the patient regarding the administration of a combination of a BRAF inhibitor and a MEK inhibitor as described herein;
In one embodiment, the treated subject became refractory to said previous treatment described herein; and in one embodiment, the treated subject became refractory to said previous treatment described herein; developed brain metastases during treatment.

Certain embodiments of the present invention are compounds of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as described herein, wherein at least one substituent comprises at least one radioactive isotope It relates to a pharmaceutical composition comprising Specific examples of radioisotopes are ^2H , ^3H , ^13C , ^14C and ^18F .

Furthermore, the present invention covers, where applicable, all optical isomers of the compounds of formula (I), i.e. diastereoisomers, diastereomeric mixtures, racemic mixtures, all their corresponding enantiomers and / or tautomers, as well as solvates thereof.

Furthermore, the present invention covers, where applicable, all optical isomers of MEK inhibitors, i.e. diastereoisomers, diastereomeric mixtures, racemic mixtures, all their corresponding enantiomers and/or tautomers. Including variants, as well as solvates thereof.

If desired, racemic mixtures of the compounds of the invention can be separated to isolate the individual enantiomers. Separation involves coupling a racemic mixture of compounds to an enantiomerically pure compound to form a mixture of diastereomers, followed by separation of the individual diastereomers by standard methods such as fractional crystallization or chromatography. It can be done by methods well known in the art, such as

In embodiments, where optically pure enantiomers are provided, optically pure enantiomers are defined as compounds containing more than 90% by weight of the desired isomer, especially 95% by weight of the desired isomer. More specifically, it means containing more than 99% by weight of the desired isomer, said weight percentage being based on the total weight of the isomers of the compound. Chirally pure or chirally enriched compounds may be prepared by chirally selective synthesis or by separation of enantiomers. Separation of enantiomers can be carried out on the final product or alternatively on a suitable intermediate.

In one embodiment, one or more additional anti-cancer agents are used in combination with a BRAF inhibitor and a MEK inhibitor as described herein, wherein the additional anti-cancer agent is a MEK degrading agent, EGFR inhibitor, EGFR degrader, inhibitor of HER2 and/or HER3, degrader of HER2 and/or HER3, SHP2 inhibitor, SHP2 degrader, Axl inhibitor, Axl degrader, ALK inhibitor, ALK degrader, PI3K inhibitors, PI3K degrading agents, SOS1 inhibitors, SOS1 degrading agents, signaling pathway inhibitors, checkpoint inhibitors, modulators of apoptotic pathways, cytotoxic chemotherapeutic agents, angiogenesis-targeted therapeutic agents, immune targeting agents, and selected from antibody-drug conjugates;

In some embodiments, one of the additional anti-cancer agents is an EGFR inhibitor. Non-limiting examples of EGFR inhibitors include cetuximab (Erbitux®), panitumumab (Vectibix®), osimertinib (merelectinib, Tagrisso®), erlotinib (Tarceva® )), gefitinib (lressa®), necitumumab (Portrazza®), neratinib (Nerlynx®), lapatinib (Tykerb®), vandetanib (Caprelsa®) and brigatinib (Alunbrig ®. Additional examples of EGFR inhibitors are known in the art.In some embodiments, the EGFR inhibitor is an allosteric EGFR inhibitor.

In some embodiments, one of the additional anticancer agents is an inhibitor of HER2 and/or HER3. Non-limiting examples of HER2 and/or HER3 inhibitors include lapatinib, canertinib, (E)-2-methoxy-N-(3-(4-(3-methyl-4-(6-methylpyridine-3 -yloxy)phenylamino)quinazolin-6-yl)allyl)acetamide (GP-724714), sapitinib, 7-[[4-[(3-ethynylphenyl)amino]-7-methoxy-6-quinazolinyl]oxy]- N-hydroxy-heptanamide (CUDC-101), mbritinib, 6-[4-[(4-ethylpiperazin-1-yl)methyl]phenyl]-N-[(1R)-1-phenylethyl]-7H- pyrrolo[2,3-d]pyrimidin-4-amine (AEE788), irbinitinib (tucatinib), poziotinib, N-[4-[1-[4-acetyl-1-piperazinyl)cyclohexyl]-4-amino- 3-pyrazolo[3,4-d]pyrimidinyl-2-methoxyphenyl]-1-methyl-2-indolecarboxamide (KIN001-111), 7-cyclopentyl-5-(4-phenoxyphenyl)-7H-pyrrolo[2 ,3-d]pyrimidin-4-ylamine (KIN001-051), 6,7-dimethoxy-N-(4-phenoxyphenyl)quinazolin-4-amine (KIN001-30), dasatinib and bosutinib.

In some embodiments, one of the additional anticancer agents is an inhibitor of SHP2. Non-limiting examples of SHP2 inhibitors include 6-(4-amino-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)pyrazin-2-amine (SHP099), [3- [(3S,4S)-4amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl]-6-(2,3-dichlorophenyl)-5-methylpyrazine-2- yl]methanol (RMC-4550), RMC-4630, TNO155, and WO2015/107493, WO2015/107494, WO2015/107495, WO2019/075265 and compounds disclosed in the brochures PCT/U82019/056786 and PCT/182020/053019.

In some embodiments, one of the additional anticancer agents is a PI3K inhibitor. Non-limiting examples include buparlisib (BKM120), alpelisib (BYL719), samotricib (LY3023414), 8-[(1R)-1-[(3,5-difluorophenyl)amino]ethyl]-N,N- Dimethyl-2-(morpholin-4-yl)-4-oxo-4H-chromene-6-carboxamide (AZD8186), tenalisib (RP6530), voxtalisib hydrochloride (SAR-245409), gedatricib (PF-05212384) ), panulisib (P-7170), tacelisib (GDC-0032), trans-2-amino-8-[4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxypyridin-3-yl)-4 -methylpyrido[2,3-d]pyrimidin-7(8H)-one (PF-04691502), duvelisib (ABBV-954), N2-[4-oxo-4-[4-(4-oxo-8-phenyl -4H-1-benzopyran-2-yl)morpholin-4-ium-4-ylmethoxy]butyryl]-L-arginyl-glycyl-L-aspartyl-L-serine acetate (SF-1126), pictilisib (GDC-0941) , 2-methyl-1-[2-methyl-3-(trifluoromethyl)benzyl]-6-(morpholin-4-yl)-1H-benzimidazole-4-carboxylic acid (GSK2636771), idelalisib (GS-1101 ), umbralisib tosylate (TGR-1202), pictilisib (GDC-0941), copanlisib hydrochloride (BAY84-1236), ductolisib (BEZ-235), 1-(4-[5-[5-amino-6 -(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl]-1-ethyl-1H-1,2,4-triazol-3-yl]piperidine-1 -yl)-3-hydroxypropan-1-one (AZD-8835), 5-[6,6-dimethyl-4-(morpholin-4-yl)-8,9-dihydro-6H-[1,4] Oxazino[4,3-e]purin-2-yl]pyrimidin-2-amine (GDC-0084), everolimus, rapamycin, perifosine, sirolimus and temsirolimus.

In some embodiments, one of the additional anticancer agents is an ALK inhibitor. Non-limiting examples include crizotinib (PF-02341066), ceritinib (LDK378), alectinib (Alecensa), brigatinib (AP26113), lorlatinib (PF-6463922), ensartinib (X-396), entrectinib (RXDX-101) , repotrectinib (TPX-0005), belizatinib (TSR-011), arcotinib (ZG-0418), foritinib (SAF-189), CEP-37440, TQ-B3139, PLB1003 and TPX-0131 be done.

In some embodiments, one of the additional anticancer agents is a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor is a CTLA-4 inhibitor, PD-1 inhibitor or PD-L1 inhibitor. In some embodiments, the CTLA-4 inhibitor is ipilimumab (Yervoy®) or tremelimumab (GP-675,206). In some embodiments, the PD-1 inhibitor is pembrolizumab (Keytruda®), nivolumab (Opdivo®) and RN888. In some embodiments, the PD-L1 inhibitor is atezolizumab (Tecentriq®), avelumab (Bavencio®) or durvalumab (Imfinzi ^™ ).

In some embodiments, one of the additional anticancer agents is an antibody-drug conjugate. Non-limiting examples of antibody-drug conjugates include gemtuzumab ozogamicin (Mylotarg™), inotuzumab ozogamicin (Besponsa®), brentuximab vedotin (Adcetris ®), ado-trastuzumab emtansine (TDM-f; Kadcyla®), mirvetuximab soravtansine (IMGN853) and anetuma vrabutansine.

In some embodiments, one of the additional anticancer agents is bevacizumab (Mvasti™, Avastin®), trastuzumab (Herceptin®), avelumab (Bavencio®), rituximab (MabThera™, Rituxan®), Edrecolomab (Panorex), Daratumumab (Darzalex®), Olaratumab (Lartruvo™), Ofatumumab (Arzerra®), Alemtuzumab (Campath®) )), cetuximab (Erbitux®), oregobomab, pembrolizumab (Keytruda®), dinutuximab (Unituxin®), obinutuzumab (Gazyva®), tremelimumab (GP-675,206), Ramucirumab (Cyramza®), Ubrituximab (TG-1101), Panitumumab (Vectibix®), Elotuzumab (EmplicitiT'V'), Necitumumab (PortrazzaT'V'), Silmutuzumab (UC-961), Ibritumomab (Zevalin®), Isatuximab (SAR650984), Nimotuzumab, Fresolimumab (GC1008), Lililumab (INN), Mogamulizumab (Poteligeo®), Ficratuzumab (AV-299), Denosumab (Xgeva®), Antibodies such as ganitumab, urelumab, pidilizumab, amatuximab, blinatumomab (AMG103; Blincyto®) or midostaurin (Rydapt).

Another embodiment of the invention is a pharmaceutical composition comprising one or more compositions, each composition comprising one or more compounds for use in accordance with the invention and one or more therapeutic agents. Pharmaceutical compositions containing inert carriers, diluents or additives are provided, as well as methods of preparing such pharmaceutical compositions. In one example, the compound of formula (I) is combined with a physiologically acceptable carrier, i.e., a carrier that is not toxic to the recipient at the doses and concentrations employed in the galenic dosage form, at an appropriate pH and desired purity. , may be formulated by mixing at ambient temperature. The pH of the formulation will depend primarily on the particular application and concentration of compound, but preferably ranges from about 3 to about 8. In one example, the compound of formula (I) is formulated in pH 5 acetate buffer. In another embodiment, the compounds of formula (I) are sterile. Compounds can be stored, for example, as solid or amorphous compositions, as lyophilized formulations, or as aqueous solutions. In one example, the MEK inhibitor is combined with a physiologically acceptable carrier, i.e., a carrier that is not toxic to the recipient at the doses and concentrations employed in galenic dosage forms, at an appropriate pH and desired purity, at ambient temperature. may be formulated by mixing at The pH of the formulation will depend primarily on the particular application and concentration of compound, but preferably ranges from about 3 to about 8. In one example, the MEK inhibitor is formulated in pH 5 acetate buffer. In another embodiment, the MEK inhibitor is sterile. MEK inhibitors can be stored, for example, as solid or amorphous compositions, as lyophilized formulations, or as aqueous solutions.

Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors to be considered in this context include the particular disorder being treated, the particular mammal being treated, the clinical presentation of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the dosing schedule, and medical treatment. There are other factors known to practitioners.

As used herein, a "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption agents. It is intended to include any and all substances compatible with the administration of pharmaceuticals, including retardants and other materials and compounds compatible with the administration of pharmaceuticals. Except insofar as any conventional media or agent is incompatible with the active compound, its use in the compositions of the invention is contemplated. Supplementary active compounds can also be incorporated into the compositions.

Pharmaceutical compositions can be obtained by treating the BRAF inhibitors and/or MEK inhibitors described herein with pharmaceutically acceptable inorganic or organic carriers or additives. Lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof, and the like can be used as carriers for tablets, coated tablets, dragees, and hard gelatin capsules, for example. Suitable carriers for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semi-solid and liquid polyols and the like. However, depending on the nature of the active substance, no carrier is usually required in the case of soft gelatin capsules. Suitable carriers for the production of solutions and syrups are, for example, water, polyols, glycerol, vegetable oils and the like. Suitable carriers for suppositories are, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols and the like.

Furthermore, the pharmaceutical composition may contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, coloring agents, flavoring agents, salts for altering the osmotic pressure, buffers, masking agents or antioxidants. can contain They can also contain still other therapeutically valuable substances.

Pharmaceutical compositions of BRAF inhibitors and MEK inhibitors, alone or in combination, contain the active ingredient(s) with the desired degree of purity in any pharmaceutically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences). 16th ed., Osol, A. (ed.) (1980)) can be prepared for storage in the form of a lyophilized formulation or an aqueous solution. Acceptable carriers, additives or stabilizers include buffers which are nontoxic to recipients at the dosages and concentrations employed and include phosphate, citrate and other organic acids; ascorbic acid and methionine. Antioxidants; preservatives (e.g. octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkylparabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt formation such as sodium. metal complexes (eg, Zn-protein complexes); and/or nonionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

Pharmaceutical compositions of BRAF inhibitors and MEK inhibitors include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. The compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well-known in the art of pharmacy. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending on the host treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of BRAF or MEK inhibitor that produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about 90 percent, preferably from about 5 percent to about 70 percent, and most preferably from about 10 percent to about 30 percent of active ingredient. Methods of preparing these compositions include the step of bringing into association a BRAF inhibitor or MEK inhibitor with the carrier and, optionally, one or more accessory ingredients. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing into association a BRAF inhibitor and a MEK inhibitor with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. can be prepared by Pharmaceutical compositions suitable for oral administration are in the form of capsules, cachets, sachets, pills, tablets, lozenges (flavor-based, usually with sucrose and acacia or tragacanth), powders, granules, or As a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as a lozenge (gelatin and glycerin, or non-preservatives such as sucrose and acacia). as an active base) and/or as a mouthwash and the like, each containing a predetermined amount of a BRAF inhibitor and a MEK inhibitor as active ingredients. BRAF inhibitors and MEK inhibitors may also be administered as a bolus, electuary or paste.

In a further embodiment of the invention the BRAF inhibitor and the MEK inhibitor are formulated in one or two separate pharmaceutical compositions.

The active ingredient can also be incorporated into colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Osol, A.; (eds.) (1980).

Formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.

The dosage can vary within wide limits and must, of course, be adjusted to the individual requirements in each concrete case. For oral administration, the dosage for adults can vary from about 0.01 mg to about 1000 mg per day of a compound of general formula (I) or a corresponding amount of a pharmaceutically acceptable solvate thereof. can. The daily dose can be administered in single or divided doses and, in addition, the upper limit can be exceeded if this proves necessary.

The following examples illustrate the invention without limiting it, but merely serve as representatives thereof. The pharmaceutical composition conveniently contains about 1-500 mg, especially 1-100 mg of the compound of formula (I). The pharmaceutical composition conveniently contains about 1-500 mg, especially 1-100 mg of the compound of formula (II). In certain embodiments, pharmaceutical compositions containing a compound of formula (I) further contain about 1-500 mg, particularly 1-100 mg of a MEK inhibitor in a fixed dose formulation.

Non-limiting examples of compositions according to the invention are:
Example A
Tablets of the following composition are manufactured by a conventional method.

Manufacturing procedure 1. Ingredients 1, 2, 3 and 4 are mixed and granulated with purified water.
2. Dry the granules at 50°C.
3. Pass the granules through suitable milling equipment.
4. Add ingredient 5, mix for 3 minutes and compress in a suitable press.

Example B-1
Prepare capsules of the following composition:

Manufacturing procedure 1. Ingredients 1, 2 and 3 are mixed in a suitable mixer for 30 minutes.
2. Add ingredients 4 and 5 and mix for 3 minutes.
3. Fill into suitable capsules.

The compound of formula (I), lactose and corn starch are first mixed in a mixer and then in a grinder. The mixture is returned to the mixer, talc is added thereto and mixed well. The mixture is filled into suitable capsules, eg hard gelatin capsules, by machine.

Example B-2
Prepare soft gelatin capsules of the following composition:

Manufacturing Procedure A compound of formula (I) is dissolved in a warm melt of the other ingredients and the mixture is filled into appropriately sized soft gelatin capsules. The filled soft gelatin capsules are processed according to normal procedures.

Example C
Prepare suppositories of the following composition:

Manufacturing Procedure The suppository mass is melted in a glass or steel vessel, mixed well and cooled to 45°C. To this is then added the finely divided compound of formula (I) and stirred until completely dispersed. The mixture is poured into suitable sized suppository molds and allowed to cool, then the suppositories are removed from the molds and individually wrapped in wax paper or metal foil.

Example D
Prepare an injection solution of the following composition:

Manufacturing Procedure A compound of formula (I) is dissolved in a mixture of polyethylene glycol 400 and water for injection (part). The pH is adjusted to 5.0 with acetic acid. Adjust the volume to 1.0 ml by adding the remaining amount of water. The solution is filtered, filled into vials using an appropriate overage, and sterilized.

Example E.
Prepare a sachet of the following composition:

Manufacturing Procedure A compound of formula (I) is mixed with lactose, microcrystalline cellulose and sodium carboxymethylcellulose and granulated with a mixture of polyvinylpyrrolidone in water. The granules are mixed with magnesium stearate and flavoring agents and filled into sachets.

Abbreviations CAS = Chemical Abstract Service; DCM = dichloromethane; DIPEA = N,N-diisopropylethylamine; DMF = dimethylformamide; DMSO = dimethylsulfoxide; -3-ethylcarbodiimide hydrochloride; ESI = electrospray ionization; EtOAc = ethyl acetate; HOOBt = 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine; LC-MS/MS = Liquid chromatography-MS/MS; MeOH = methanol; MS = mass spectrometry; NMP = N-methyl-2-pyrrolidone; PCR = polymerase chain reaction; rt = room temperature;

Test agent (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1-sulfonamide ( Referred to herein as Compound Ia) was supplied by Roche, Basel, Switzerland as a powder and resuspended prior to use. 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl]methyl]benzamide sodium salt ( Referred to herein as compound IIa) was provided by Chugai, Tokyo, Japan as a powder. Cobimetinib (catalog number HY-13064A), encorafenib (catalog number HY-15605) and binimetinib (catalog number HY-15202) were purchased from MedChemExpress.

Cell lines and culture conditions Cell lines were obtained from ATCC and maintained in a humidified incubator with 5% CO2 under standard conditions and passaged twice weekly. Culture conditions are reported in the table below:

animal:
For in vivo studies, female mice aged 7-9 weeks (at the start of experiments) were purchased from Charles River Laboratories. The strain used in the experiment was CB. 17 SCID.

For xenograft establishment, cells were suspended in medium consisting of 50% Matrigel and 50% Hank's balanced salt solution and injected subcutaneously into the right flank. Mice were randomized prior to treatment when tumor volumes reached approximately 100 mm ³ .

EXAMPLES The following examples and figures are provided to illustrate the invention and have no limiting character.

Example 1
The A375 cell line, which exhibits BRAF V600E and the RAF dimer-inducing mutation NRAS Q61K, was utilized to model resistance mechanisms to BRAFi and BRAFi/MEKi combinations. A375 NRAS Q61K cells were treated with either compound Ia, encorafenib alone, or in combination with 10 nM MEKi cobimetinib for 1 hour and then utilized for Western blot analysis of phosphorylated ERK, also called P-ERK (Figure 1). Similarly, A375 NRAS Q61K cells were seeded at 500 cells/well, treated with either compound Ia, encorafenib alone or in combination with MEKi cobimetinib and incubated for 12 days. The resulting colonies were fixed and stained with a solution of crystal violet/methanol (Fig. 2). According to its paradox-breaking properties, Compound Ia drives superior P-ERK inhibition compared to encorafenib and correspondingly the combination of Compound Ia with cobimetinib is induced by the encorafenib/cobimetinib combination. In comparison, it provides superior P-ERK suppression.

Example 2
Immunodeficient mice were implanted with the cell line A375 NRAS, which exhibits BRAF V600E and the RAF dimer-inducing mutation NRAS Q61K, as a model of resistance to first-generation BRAFi and BRAFi/MEKi. Once tumors were established (100 mm3), mice were randomized to receive either Compound Ia (20 mg/kg), its combination with MEKi cobimetinib (5 mg/kg, QD PO) or cobimetinib alone once daily (QD). It was administered orally (PO) (Fig. 3). The same mouse model was also treated once daily with compound Ia (20 mg/kg) in combination with a different MEK inhibitor binimetinib (10 mg/kg, BID PO) (Figure 4). For comparison, one animal in the experimental arm was treated with the paradox-inducing BRAFi encorafenib (36 mg/kg, QD PO) in combination with binimetinib, an FDA-approved combination for the treatment of metastatic melanoma. treated. The same mouse model was also treated once daily with Compound Ia (20 mg/kg) in combination with a different MEK inhibitor Compound IIa (0.0625 mg/kg in one cohort and 1.0 mg/kg in the second cohort, PO). treated (Fig. 5).

Example 3
A375 cell line was obtained from ATCC and maintained in a humidified incubator with 5% CO2 under standard conditions. Cells were treated with encorafenib and compound IIa at the indicated concentrations for 7 days in 384-well plates (U-bottom). Cell viability was measured by CellTiter-Glo 2.0 (Promega, G9243) and EnVision plate reader (Perkin Elmer). Cell growth inhibition by compounds was calculated by the formula (1−(T−V0)/(V−V0))×100 (%), where T represents the measured value of wells containing compound and V is compound represents measurements of wells without , and V0 represents measurements of wells without cells. The 80% inhibitory concentration (IC80) values were calculated and the IC80 isobolograms were plotted. The X and Y axes represent the concentrations of compound IIa and encorafenib, respectively. The isobol is hyperbolic and lies below the additivity line (dashed line), indicating synergy between encorafenib and compound IIa (Figure 6).

Claims (24)

A pharmaceutical composition for treating or preventing cancer comprising a combination of a BRAF inhibitor and a MEK inhibitor, wherein the BRAF inhibitor is represented by formula (I):

or a pharmaceutically acceptable salt or solvate thereof ,
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] A pharmaceutical composition selected from methyl]benzamide, cobimetinib, trametinib and binimetinib, or a pharmaceutically acceptable salt thereof . A pharmaceutical composition for treating or preventing cancer in combination with a MEK inhibitor, comprising a BRAF inhibitor, wherein the BRAF inhibitor has formula (I):

or a pharmaceutically acceptable salt or solvate thereof ,
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] A pharmaceutical composition selected from methyl]benzamide, cobimetinib, trametinib and binimetinib, or a pharmaceutically acceptable salt thereof . A pharmaceutical composition for treating or preventing cancer in combination with a BRAF inhibitor, comprising a MEK inhibitor, wherein the BRAF inhibitor has formula (I):

or a pharmaceutically acceptable salt or solvate thereof ,
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] A pharmaceutical composition selected from methyl]benzamide, cobimetinib, trametinib and binimetinib, or a pharmaceutically acceptable salt thereof . The compound of formula (I) is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine -1-sulfonamide, the pharmaceutical composition according to any one of claims 1 to 3. the BRAF inhibitor is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazolin-6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1 - a pharmaceutical composition according to any one of claims 1 to 3, which is a sulfonamide. The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] 4. The pharmaceutical composition according to any one of claims 1 to 3, which is methyl]benzamide or a pharmaceutically acceptable salt thereof. The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] 4. The pharmaceutical composition according to any one of claims 1 to 3, which is methyl]benzamide sodium salt. 4. The pharmaceutical composition of any one of claims 1-3, wherein the MEK inhibitor is cobimetinib or a pharmaceutically acceptable salt thereof. The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] 6. The pharmaceutical composition of claim 5, which is methyl]benzamide sodium salt. 6. The pharmaceutical composition of claim 5, wherein said MEK inhibitor is cobimetinib or a pharmaceutically acceptable salt thereof. 4. Any of claims 1 to 3 for melanoma or non-small cell lung cancer, wherein an effective amount of the pharmaceutical composition of any of claims 1 to 3 is administered to a patient in need thereof. or the pharmaceutical composition according to claim 1. 4. The pharmaceutical composition of any one of claims 1-3, wherein the BRAF inhibitor and the MEK inhibitor are both orally administered. 4. The pharmaceutical composition of any one of claims 1-3, wherein the BRAF inhibitor is administered concurrently with the MEK inhibitor. 4. The pharmaceutical composition of any one of claims 1-3, wherein said BRAF inhibitor and said MEK inhibitor are co-formulated. 4. The pharmaceutical composition of any one of claims 1-3, wherein said BRAF inhibitor is administered sequentially with said MEK inhibitor. 4. The pharmaceutical composition according to any one of claims 1 to 3, wherein said cancer is thyroid cancer, colorectal cancer, melanoma, brain cancer or non-small cell lung cancer. 4. The pharmaceutical composition of any one of claims 1-3, wherein the cancer is associated with BRAF ^V600 mutations. 4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the cancer is BRAF ^V600 mutation-positive unresectable or metastatic cancer. BRAF ^V600 mutations are determined by (a) PCR or sequencing nucleic acid (e.g., DNA) extracted from a sample of patient tumor tissue and/or body fluid; and (b) expression of BRAF ^V600 in said sample. 4. The pharmaceutical composition of any one of claims 1-3, determined using a method comprising determining the MEK degrading agents, EGFR inhibitors, EGFR degrading agents, HER2 and/or HER3 inhibitors, HER2 and/or HER3 degrading agents, SHP2 inhibitors, SHP2 degrading agents, Axl inhibitors, Axl degrading agents, ALK inhibitors, ALK degrading agents, PI3K inhibitors, PI3K degrading agents, SOS1 inhibitors, SOS1 degrading agents, signal transduction pathway inhibitors, checkpoint inhibitors, regulators of apoptotic pathways, cytotoxic chemotherapeutic agents, angiogenesis targeting therapeutic agents, 4. The pharmaceutical composition according to any one of claims 1 to 3, comprising one or more additional anti-cancer agents selected from immunotargeting agents, as well as antibody-drug conjugates. A pharmaceutical composition for treating or preventing cancer, in combination with a MEK inhibitor, comprising formula (I):

or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable additives ,
The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] A pharmaceutical composition selected from methyl]benzamide, cobimetinib, trametinib and binimetinib, or a pharmaceutically acceptable salt thereof . The compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof is (3R)-N-[2-cyano-4-fluoro-3-(3-methyl-4-oxo-quinazoline- 22. The pharmaceutical composition of claim 21 , which is 6-yl)oxy-phenyl]-3-fluoro-pyrrolidine-1-sulfonamide. The MEK inhibitor is 2-(4-cyclopropyl-2-fluoroanilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)pyridin-4-yl] 23. The pharmaceutical composition according to claim 21 or 22 , which is methyl]benzamide sodium salt. 23. The pharmaceutical composition of claim 21 or 22 , wherein said MEK inhibitor is cobimetinib or a pharmaceutically acceptable salt thereof.

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