Pharmaceutical combinations for the treatment of melanoma

Abstract

Pharmaceutical compositions and methods for treating metastatic melanoma and resistant BRAF-mutated melanoma are provided. [Solution] The present invention relates to a combination pharmaceutical comprising a cyclin-dependent kinase (CDK) inhibitor represented by the compound of the following formula I or a pharmaceutically acceptable salt thereof and at least one anti-cancer agent selected from a BRAF inhibitor or a MEK inhibitor for use in the treatment of melanoma. The present invention also relates to a method for treating melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of a CDK inhibitor and a therapeutically effective amount of at least one anti-cancer agent selected from a BRAF inhibitor or a MEK inhibitor. JPEG2022068335000020.jpg4942

Description

[Technical Field]

[0001] The present invention provides compounds of formula I (as described herein) for use in the treatment of melanoma. CDK (cyclin-dependent kinase) compounds represented by the compounds of (i) or pharmaceutically acceptable salts thereof inhibitors of BRAF (serine-threonine protein kinase B-raf ) inhibitors or MEK (mitogen-activated protein kinase) inhibitors The present invention also relates to a combination drug containing at least one anticancer drug. The present invention also relates to a method for treating melanoma using the same. [Background technology]

[0002] Melanoma is the most serious type of skin cancer. In fact, melanoma is caused by the action of melanocytes and It is a malignant tumor that begins in cells called melanocytes, which are pigment-producing cells. Melanoma occurs most often in the skin, but can also occur in the eye or on the lining of the nose, mouth, or genitals. Melanoma can then spread to the internal organs. If melanoma begins on the skin: It is called cutaneous melanoma. When melanoma occurs in the eye, it is called ocular melanoma or intraocular melanoma. The incidence of melanoma is increasing worldwide, and malignant melanoma accounts for 80% of skin cancer deaths. It has been reported that this is the cause (N.Engl.J.Med., 2010, No. 36 3, No. 8, pp. 711-723).

[0003] When melanoma spreads, cancer cells are often found in the lymph nodes. If the nodes have been reached, this means that the cancer cells have spread to other parts of the body, such as the liver, lungs, or brain. This is a sign that the cancer has spread to other organs, potentially causing metastatic melanoma. In fact, melanoma metastasis is extremely aggressive, and the survival time of patients with metastatic melanoma is, on average, Unfortunately, there is no effective treatment for metastatic melanoma. Diagnosis and prompt surgical resection are possible options for patients with a potential cure. Treatment regimens for metastatic melanoma initially consist of interleukin-2, dacarbazine, and tetanus. It consists of drugs such as mozolomide, fotemustine, and carboplatin, respectively. It is associated with poor response rates and poor overall survival. For example, dacarbazine has a response rate of 7-12%. The response rate and overall survival time were found to be 5 to 8 months after the start of treatment (N.En gl. J. Med., 2011, Vol. 364, No. 26, pp. 2507-2516).

[0004] Mitogen-activated protein (MAP) kinase activity in a large panel of common cancers Studies of enzyme pathways have shown that 40-60% of melanomas contain serine-threonine protein kinases Carrying an activating mutation in the gene encoding B-raf (BRAF) Among the BRAF mutations observed in melanoma, over 90% occur at codon 600. Among these, over 90% are single nucleotide mutations, resulting in , which results in a substitution of glutamic acid for valine (BRAF V600E). The mutation is BRAF V600K, which causes a substitution of valine for lysine, and is responsible for the 5-6% risk of melanoma. % of the total, followed by BRAF V600R (valine to arginine substitution) and BR AF V600D (valine replaced by aspartic acid) (Journal of T Translational Medicine, 2012, Vol. 10, No. 85, 1-9 page).

[0005] BRAF inhibitors, at low nanomolar concentrations, are effective in the majority of patients with BRAF-mutant melanoma It has been found to be effective in shrinking tumors in patients with rheumatoid arthritis. Factors limiting efficacy are drug resistance and Progression-free survival is limited to 5-7 months. Some examples include BAY43-9006 (sorafenib, Bayer), vemurafenib PLX4032, Plexxikon; RG7204, RO5185426, Hof mann-LaRoche), GDC-0879 (GlaxoSmithKline), Dabrafenib (GSK2118436, GlaxoSmithKline), PLX4 720 (Hofmann-LaRoche), BMS-908662 (XL281, Br istol-Myers Squibb), LGX818 (Novartis), PLX 3603 (RO5212054, Hofmann-LaRoche), ARQ-736 ( ArQule), DP-4978 (Deciphera) and RAF265 (Nova) rtis) are examples.

[0006] Vemurafenib is a potent inhibitor of mutant BRAF, particularly the BRAF V600E mutation. Vemurafenib is effective in patients with melanoma harboring the BRAF V600E mutation. The response was observed in the bone, liver, and small intestine. It was observed in all disease sites, including the intestine. After an early response, responsive tumors were found to develop resistance to treatment. In some patients with the V600E mutation, tumors develop resistance without evidence of an initial response. (N.Engl.J.Med., 2010, Vol.363, No.9, pp.809-819) Dabrafenib inhibits human wild-type BRAF and CRAF enzymes as well as mutant BRAF and CRAF enzymes. The strong BRAF V600E, BRAF V600K, and BRAF V600D variants It is a potent and selective RAF kinase inhibitor.

[0007] MEK1 and MEK2 bind to both tyrosine and threonine phosphates within target proteins. Dual specificity kinases are protein serine / triphosphate kinases that catalyze the synthesis of ATP. It is part of the oncogenic kinase family. Protein phosphorylation is used in signal transduction. It is the most widespread class of post-translational modifications (Biochemical and Biop Physical Research Communications, 2012, No. 4 17, pp. 5-10). MEK1 and MEK2 are RAS-RAF-MEK-ER K signaling cascade (sometimes called mitogen-activated protein kinase (MAPK) ubiquitously expressed hydrophilic non-receptor molecules that participate in the cascade of Ras-mediated Raf activation is mediated by the activation of MEK1 and MEK2 (MAP kinases). / ERK kinase 1 and 2), which in turn activates tyrosine 185 and ERK1 and ERK2 (extracellular signal-regulated kinases 1 and 2) on leonine 183 Activated ERK1 and ERK2 translocate and accumulate in the nucleus, where they phosphorylate ERK1 and ERK2. In vivo, it phosphorylates various substrates, including transcription factors that control cell proliferation and survival. The controlled regulation of these cascades is involved in cell proliferation and differentiation. In contrast, unregulated activation of these kinases can lead to carcinogenesis. Dysregulation of the S / RAF / MEK pathway has been detected in over 30% of human tumors, but M Mutations in the EK1 and MEK2 genes were rarely detected, and therefore MEK1 / 2 hyperactivation is usually due to gain-of-function mutations in RAS and / or BRAF Considering the importance of the Ras / Raf / MEK / ERK pathway in human cancer development and kinase components of signaling cascades in cancer and other proliferative diseases. It is a potentially important target for modulating disease progression.

[0008] Some examples of potent MEK inhibitors include trametinib (Mekinist™) ), selumetinib (AstraZeneca), binimetinib (Array Bioph arma), PD-0325901 (Pfizer), cobimetinib (Exelixis ), refametinib (Valeant Pharmaceutical Int.), Macerutib (Santhera Pharmaceuticals), TAK-733 ( Takeda) and WX-554 (UCB Pharma SA).

[0009] Trametinib is a potent and selective inhibitor of MEK1 and MEK2. BRAF V600E or BRAF V600K mutations not previously treated with Trametinib compared with chemotherapy in patients with metastatic melanoma Significant clinical activity has been demonstrated with either dacarbazine or paclitaxel. Reka). Summary of the Invention [Problem to be solved by the invention]

[0010] Thus, metastatic melanoma and resistant BRAF-mutant melanoma are difficult to treat with existing therapies. It is clear from the above discussion that it continues to be difficult to New effective drugs for these conditions are being developed, both to prevent the disease and to treat the condition. There continues to be a need for effective treatments. [Means for solving the problem]

[0011] In one aspect, the present invention provides a compound of formula I (as described herein) for use in the treatment of melanoma. CDKs (as shown) or pharmaceutically acceptable salts thereof cyclin-dependent kinase (BRAF) inhibitors and BRAF (serine-threonine protein kinase B-raf inhibitors or MEK (mitogen-activated protein kinase) inhibitors and at least one anticancer agent selected from the group consisting of:

[0012] In another aspect, the present invention provides a compound or drug of Formula I (as described herein): A therapeutically effective amount of a CDK inhibitor, represented by a physiologically acceptable salt thereof, is administered to a subject in need of a BRAF inhibitor. and a therapeutically effective amount of at least one anticancer drug selected from the group consisting of a steroid drug and a MEK inhibitor. and administering to a subject in need thereof.

[0013] In another aspect, the present invention provides a compound selected from the group consisting of compounds of formula I: and at least one CDK inhibitor selected from a BRAF inhibitor or a MEK inhibitor. an anti-cancer drug and one or more pharmaceutically acceptable carriers, excipients or additives, The present invention relates to a pharmaceutical composition.

[0014] In a further aspect, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof for the treatment of melanoma. CDK inhibitors, represented by acceptable salts thereof, and BRAF or MEK inhibitors and at least one anticancer agent selected from the group consisting of:

[0015] In a further aspect, the present invention provides a compound of formula I for the manufacture of a medicament for the treatment of melanoma. and BRAF inhibitors, which are represented by the compounds of formula (I) and (II) or pharmaceutically acceptable salts thereof. The present invention relates to the use of at least one anticancer agent selected from the group consisting of a steroid inhibitor and a MEK inhibitor.

[0016] In a further aspect, the present invention provides a compound selected from the group consisting of compounds of formula I or a pharmaceutically acceptable salt thereof: and at least one selected from a BRAF inhibitor or a MEK inhibitor. and an anti-cancer agent.

[0017] Other aspects and further scope of applicability of the present invention will become apparent from the detailed description that follows. Let's become happy. [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 depicts the effects of Compound A (voruciclib) and vemurafenib, alone and in combination, on cell cycle and apoptosis in G361 melanoma cells after 5 days using flow cytometry. [Figure 2] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib, alone and in combination, on early apoptosis in G361 melanoma cells treated for 24 hours using Annexin staining. [Figure 3]FIG. 1 depicts the effect of Compound A (voruciclib) and vemurafenib, alone and in combination, on the cell cycle and apoptosis of SK-MEL3 melanoma cells after 5 days using flow cytometry. [Figure 4] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib (48h) alone and in combination in A375 cell line. [Figure 5] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib (48h) alone and in combination in G361 cell line. [Figure 6] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib (48h) alone and in combination in the MDA-MB435S cell line. [Figure 7a] 1 is a graph depicting dose response curves for Compound A (voruciclib) and vemurafenib alone (48 h) on the A375 parental cell line and the A375R resistant cell line. [Figure 7b] 1 is a graph depicting dose response curves for Compound A (voruciclib) and vemurafenib alone (48 h) on the A375 parental cell line and the A375R resistant cell line. [Figure 8a] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib (48h) alone and in combination in A375 resistant cell lines. [Figure 8b] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib (48h) alone and in combination in A375 resistant cell lines. [Figure 9a] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib (48h) alone and in combination in the A375R resistant cell line. [Figure 9b] 1 is a graph depicting the effect of Compound A (voruciclib) and vemurafenib (48h) alone and in combination in the A375R resistant cell line. [Figure 10a]1 is a graph depicting the effect of Compound A (voruciclib) and trametinib (48h) alone and in combination in A375 resistant cell lines. [Figure 10b] 1 is a graph depicting the effect of Compound A (voruciclib) and trametinib (48h) alone and in combination in A375 resistant cell lines. [Figure 11a] 1 is a graph depicting the effect of Compound A (voruciclib) and trametinib (48h) alone and in combination in the A375R resistant cell line. [Figure 11b] 1 is a graph depicting the effect of Compound A (voruciclib) and trametinib (48h) alone and in combination in the A375R resistant cell line. [Figure 12a] 1 is a graph depicting the effect of Compound A (voruciclib) and dabrafenib (48h) alone and in combination in A375 resistant cell lines. [Figure 12b] 1 is a graph depicting the effect of Compound A (voruciclib) and dabrafenib (48h) alone and in combination in A375 resistant cell lines. [Figure 13a] 1 is a graph depicting the effect of Compound A (voruciclib) and dabrafenib (48h) alone and in combination in the A375R resistant cell line. [Figure 13b] 1 is a graph depicting the effect of Compound A (voruciclib) and dabrafenib (48h) alone and in combination in the A375R resistant cell line. DETAILED DESCRIPTION OF THE INVENTION

[0019] The general terms used hereinbefore and hereinafter are: Within the context of this disclosure, unless otherwise indicated, preferably have the following meanings: Therefore, definitions of general terms as used in the context of the present invention are provided herein below. R:

[0020] The singular forms "a," "an," and "the" are used unless the context clearly dictates otherwise. Includes multiple references unless otherwise noted.

[0021] The use of "(s)" as part of a term refers to the term individually or to the plural. For example, the term drug(s) may refer to a single drug or multiple drugs. It shows.

[0022] As used herein, the term "at least one" refers to one or more For example, the term "at least one anti-cancer agent" means that the combination may include a single anti-cancer agent or or multiple anticancer drugs.

[0023] "Substituted" or "substituted with" means that such substitution is at the substituted atom and This includes the implicit requirement that the allowed valence of the substituents be met, as well as rearrangements, cyclizations, decompositions, and It should be understood that "stable" means a compound that does not readily undergo transformations such as isolation.

[0024] As used herein, the term "pharmaceutically acceptable" refers to a carrier, excipient, additive, or The agent and / or salt must be compatible with the other ingredients of the formulation and be suitable for the recipient. "Pharmaceutically acceptable" also means that a composition or or dosage form is within the scope of sound medical judgment and is not likely to result in excessive toxicity, irritation, allergic reaction, or Suitable and acceptable for animal or human use without adverse or other problems or complications This means that the benefit / risk ratio is justified.

[0025] As used herein, the term "combination" or "combination drug" refers to a combination of anti-cancer drugs. Combination administration, in the context of the present invention, is the combination of a CDK inhibitor (a compound of Formula I) with a BRAF inhibitor or It means the combination administration of at least one anticancer drug selected from the MEK inhibitors. Anticancer drugs may be used independently, simultaneously, or with combination partners that are particularly capable of exhibiting synergistic effects. The doses can be administered separately at a time interval that allows for a

[0026] Cyclin-dependent kinases (CDKs) are activated at specific stages of the cell cycle CDKs are a family of enzymes. They consist of a catalytic subunit (the actual cyclin-dependent kinase It consists of at least nine C DK(CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CD K8, CDK9, etc.) and at least 15 different types of cyclins (cyclins A, B1, B2, D1, D2, D3, E, H, etc. Each step of the cell cycle is Regulated by such CDK complexes: G1 / S transition (CDK2 / cyclin A , CDK4 / cyclin D1-D3, CDK6 / cyclin D3), S phase (CDK2 / S icrin A), G2 phase 30 (CDK1 / cyclin A), G2 / M transition phase (CDK1 / cyclin B).

[0027] As used herein, the term "CDK inhibitor" refers to an inhibitor of one or more cyclins. These drugs are capable of inhibiting cytokine-dependent kinase(s) (CDKs). Aberrant expression and overexpression of kinases has been demonstrated in many disease states, including cancer. In the context of the present invention, the CDK inhibitor contained in the pharmaceutical combination of the present invention is The compounds of the present invention are compounds of Formula I or pharmaceutically acceptable salts thereof. CDK1 / cyclin B, CDK2 / cyclin E, CDK4 / cyclin D, CD K4 / cyclin D1 and / or CDK9 / cyclin T1.

[0028] As used herein, the term "BRAF inhibitor" refers to an inhibitor of BRAF kinase or Mutated BRAF kinase activity (one or more mutant forms of the serine-threonine protein It refers to drugs that can inhibit the protein kinase B-RAF (BRAF). 90 percent of the mutations result in a substitution of glutamic acid for valine at amino acid 600 (V600E mutation). Therefore, the term "BRAF inhibitor" is within its scope. , a compound capable of inhibiting BRAF or a mutated form thereof; or a V600 mutated form of BRAF Compounds capable of inhibiting F; or the V600E mutation in both non-refractory and refractory melanoma The present invention encompasses compounds capable of inhibiting any of the forms of BRAF.

[0029] As used herein, the term "MEK inhibitor" refers to a compound that inhibits the action of mitogen-activated proteins. Drugs that can interact with MEK and inhibit its enzymatic activity Inhibiting MEK enzyme activity, in turn, phosphorylates the substrate peptide or protein. MEK1 and MEK2 reduce the ability of MEK to oxidize RAS-RAF-MEK. It is a protein kinase involved in the K-ERK signaling cascade. Cades mediate a wide range of events, including apoptosis, cell cycle progression, cell migration, differentiation, metabolism, and proliferation. MEK inhibitors are involved in the regulation of a variety of processes. Included within the scope of the present invention are compounds capable of inhibiting MEK.

[0030] As used herein, the terms "synergistic" or "synergistic effect" or "synergistic action" The term, as used herein, refers to the therapeutic effect of a combination of compounds (BRAF inhibitors and and CDK inhibitors, i.e., compounds of formula I), which may enhance the additive effects of compounds used in combination medicines. Advantageously, such synergistic effects between the active ingredients (therapeutically active compounds) are When combined, these allow for the use of smaller doses of one or both active ingredients, Greater efficacy at the same dose and / or preventing or slowing the development of multidrug resistance The combination index (CI) method of Chou and Talalay was used to The CI value can be used to determine the synergistic, additive, or antagonistic effects of the compounds used. If the CI is less than 1, synergy exists between the compounds used in this combination, and the CI value is 1. If they are equal, there is an additive effect between the compounds used in this combination, and the CI value is greater than 1. In this case, an antagonistic effect exists. A synergistic effect occurs when the compounds contained in the pharmaceutical combination or the present invention The compounds can be administered simultaneously by co-forming the compositions and via a unit dosage form. or as separate formulations administered simultaneously or sequentially. This can be achieved.

[0031] As used herein, "melanoma" refers to a tumor arising in the melanocytic system of the skin and other organs. It refers to a condition characterized by the growth of tumors caused by melanocytes. Most melanocytes are found in the skin. Melanoma occurs in the skin, but has also been found in the meninges, gastrointestinal tract, lymph nodes, and eyes. If it occurs in the eyes, it is called cutaneous melanoma. Melanoma can also occur in the eyes, which Melanoma can occur in the meninges, gastrointestinal tract, lymph nodes, or melanin-producing areas. It occurs rarely in other areas where formative cells are found.

[0032] The terms "variant melanoma" or "malignant melanoma," used interchangeably, are Refers to a neoplasm of melanocytes containing melanoma cells with defects (also called "mutations") Malignant melanoma usually arises from or near a nevus and has a marked tendency to metastasize. It consists of a mass of cells. 40-60% of melanomas are caused by the serine-threonine protein kinase B -carry an activating mutation in the gene encoding RAF (BRAF) Among the BRAF mutations observed in melanoma, over 90% are at codon 600. Among these, over 90% contain a single nucleotide that results in the substitution of valine with glutamic acid. The second most common mutation is BRAF V600E. The BRAF V600K mutation causes a substitution of lysine for BRAF V600K, which accounts for 5-6% of melanomas. This is followed by the BRAF V600R and BRAF V600D. (Journal o f Translational Medicine, 2012, Vol. 10, No. 85, (Pages 1-9).

[0033] The term "metastatic melanoma" refers to the spread of cancer beneath the surface of the skin via the lymphatic system and / or blood vessels. Other parts of the body, including the subcutaneous tissue located in the lungs, lymph nodes, and other organs, e.g., lungs, liver Stage III melanoma refers to melanoma that has spread to the liver, bones, or brain. There is no evidence of distant metastasis. Stage IV melanoma is characterized by the location and severity of distant metastases. This stage is characterized by serum lactate dehydrogenase (LDH) levels. Also called metastatic melanoma.

[0034] Unless otherwise noted, the term "melanoma" also refers to recurrent or resistant melanoma. The terms "recurrent" or "resistant" refer to repeated outgrowth of melanoma or melanoma. This refers to the progression of a tumor, regardless of whether the disease was cured prior to said proliferation or progression. do not have.

[0035] Thus, combination pharmaceuticals (as described herein) are provided for the treatment of melanoma. Treatment is for non-refractory, metastatic, or refractory (resistant) BRAF-mutant melanoma, particularly BRAF It refers to the treatment of V600 mutant melanoma, more specifically BRAF V600E mutant melanoma. vinegar.

[0036] The term "non-responsive / refractory" when used in relation to melanoma refers to the progression of melanoma treatment. To address this issue, currently available cancer therapies, such as chemotherapy, radiation therapy, surgery, and hormone therapy, and / or subjects with melanoma who are being treated with biotherapy / immunotherapy, etc., or As used herein to refer to a patient, the therapy is clinically appropriate to treat the patient. These patients need additional effective therapy, i.e., therapy The phrase also refers to a person who responds to therapy but suffers from side effects, relapses, or other conditions. develop resistance or experience any alleviation of one or more symptoms of melanoma. In various embodiments, "non-responsive / refractory" may refer to a subject or patient who is not responding to treatment. This means that at least some significant portion of the cancer cells have not died, or that these cells have not died. This means that the cancer cells are not resistant to treatment. The determination is made using the art-accepted meaning of "refractory" in this context. , by any method known in the art for assaying the effectiveness of a treatment on cancer cells in This can be done either in vivo or in vitro.

[0037] As used herein, the term "treatment cycle" refers to the administration of a CDK inhibitor, i.e., a compound or a pharmaceutically acceptable salt thereof; and a BRAF inhibitor or a MEK inhibitor It refers to the time period during which repeated successive administrations of at least one selected anticancer drug are carried out. vinegar.

[0038] The term "apoptosis" refers to the natural process of programmed cell death Apoptosis is a process in which a cell uses specialized cellular mechanisms to kill itself. The process of destruction involves the breakdown of cells into membrane-bound particles, which are then transported by phagocytes. Apoptosis is a process by which metazoans control cell number and ensure the survival of the animal. It is a mechanism that allows threatening cells to be eliminated.

[0039] The term "subject" as used herein means an individual who has been the object of treatment, observation or experiment. The present invention relates to an animal, preferably a mammal, most preferably a human, that has been treated with melanoma. The term "subject" is used interchangeably with the term patient. In the context of the present invention, the phrase "subject in need thereof" includes variants or It means a subject in need of treatment for malignant melanoma. The phrase "melanoma" refers to a subject (patient) diagnosed with variant or malignant melanoma.

[0040] The term "mammal" as used herein refers to humans as well as non-human mammals. Non-human mammals are intended to include, but are not limited to, domestic animals, e.g. For example, cows, pigs, horses, dogs, cats, rabbits, rats and mice, as well as non- Poultry animals are included.

[0041] The term "therapeutically effective amount" as used herein refers to the amount of a substance that is required for such treatment. a CDK inhibitor that is sufficient to provide a therapeutic benefit when administered to a subject, i.e., A compound of formula I or a pharmaceutically acceptable salt thereof, and a BRAF inhibitor or a MEK inhibitor The therapeutic benefit refers to the amount of at least one anti-cancer agent selected from the group consisting of: (i) preventing or delaying one or more symptoms of melanoma; (ii) preventing or delaying one or more symptoms of melanoma; (iii) ameliorating or eliminating one or more symptoms of melanoma; or do.

[0042] "Treating" melanoma in a subject, preferably a mammal, more preferably a human " or "treatment" or "treated" includes: (i) inhibition of melanoma (ii) inhibiting the onset of melanoma; (iii) reducing the regression of melanoma; (iii) inhibiting tumor cell invasion into peripheral organs; (iv) inhibiting metastasis ( (v) melanoma remission, i.e., a decrease, slowing down, or complete cessation; (vi) reducing the severity of symptoms associated with melanoma; and (vi) reducing one or more of the symptoms associated with melanoma. relieves some of the symptoms.

[0043] According to one aspect of the present invention, there is provided a compound of formula I for use in the treatment of melanoma.

[0044] [ka] (wherein Ar is a phenyl group, and this phenyl group is selected from chlorine and trifluoromethyl) substituted with one or two different substituents selected from or a pharmaceutically acceptable salt thereof, and a BRAF inhibitor. and at least one anticancer drug selected from the group consisting of a steroid drug, a steroid inhibitor, and a MEK inhibitor. It is served.

[0045] According to another embodiment, the CDK inhibitor contained in the pharmaceutical combination of the present invention is a compound represented by formula I ( wherein Ar is substituted with two different groups selected from chlorine and trifluoromethyl. wherein R is a phenyl group, or a pharmaceutically acceptable salt thereof.

[0046] According to another further embodiment, the CDK inhibitor contained in the pharmaceutical combination of the present invention is Compounds of formula I (wherein Ar is a phenyl group substituted with chlorine) or pharmaceutically It is selected from acceptable salts thereof.

[0047] Preparation of a compound of formula I or a pharmaceutically acceptable salt thereof, and preparation of a pharmaceutical composition containing said compound The preparation of pharmaceutical compositions is described in PCT Patent Publication WO2004004632 (U.S. Patent No. 7,271, 193) and PCT Patent Publication WO2007148158. These PCT patent publications disclose that compounds of Formula I may be used to treat proliferative disorders. As shown hereinabove, the compound of formula I can be The compound of formula I can be used in the form of a salt thereof. Preferred salts of the compound of formula I include acetate, alkane, methyl ... Glycates, ascorbic acids, aspartates, benzoates, benzenesulfonates, Bisulfate, borate, cinnamate, citrate, ethanesulfonate, fumarate, glutamate Chlorate, glutamate, glycolate, hydrochloride, hydrobromide, hydrofluoride , ketoglutarate, lactate, maleate, malonate, mesylate, nitrate, oxalate Salicylate, pamoate, perchlorate, phosphate, picrate, salicylate, succinate, sulfonate sulfamate, sulfate, tartrate, tosylate, trifluoroacetate and salts known to those skilled in the art Other acid addition salts are known.

[0048] In one embodiment, the CDK inhibitor (compound of formula I) contained in the pharmaceutical combination is (+)- trans-2-(2-chloro-4-trifluoromethylphenyl)-5,7-dihydro 8-(2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chromene- 4-one or a pharmaceutically acceptable salt thereof.

[0049] In another embodiment, the CDK inhibitor (compound of formula 1) contained in the pharmaceutical combination is (+) -trans-2-(2-chloro-4-trifluoromethylphenyl)-5,7-dihydriodide hydroxy-8-(2-hydroxymethyl-1-methylpyrrolidin-3-yl)-chromene- 4-one hydrochloride (referred to herein as "Compound A). Compound A is also referred to herein as "bo (also known as lucilib).

[0050] In one embodiment, the CDK inhibitor (compound of formula 1) contained in the pharmaceutical combination is (+)- trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydro (1-methyl-pyrrolidin-3-yl)-chromen-4-one or its pharmaceutical composition It is a commercially acceptable salt.

[0051] In another embodiment, the CDK inhibitor (compound of formula 1) contained in the pharmaceutical combination is (+) -trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxy (Honmei)-1-methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride Compound B is also referred to herein as "ribiciclib." (Can be).

[0052] In one embodiment, the BRAF inhibitor contained in the pharmaceutical combination is a V600 mutant form of B It is an inhibitor of RAF.

[0053] In one embodiment, the BRAF inhibitor contained in the pharmaceutical combination is a BRAF inhibitor of the V600E mutant form. It is an inhibitor of BRAF.

[0054] In one embodiment, the BRAF inhibitor contained in the combination pharmaceutical is BAY43-9006 (sorafenib, Bayer), vemurafenib (PLX4032, Plexxikon ;RG7204, RO5185426, Hofmann-LaRoche), GDC-0 879 (GlaxoSmithKline), dabrafenib (GSK2118436, GlaxoSmithKline), PLX4720 (Hofmann-LaRoche ), BMS-908662 (XL281, Bristol-Myers Squibb) , LGX818 (Novartis), PLX3603 (RO5212054, Hofm ann-LaRoche), ARQ-736 (ArQule), DP-4978 (Dec. iphera) or RAF265 (Novartis).

[0055] In one embodiment, the BRAF inhibitor contained in the pharmaceutical combination is vemurafenib.

[0056] In one embodiment, the BRAF inhibitor contained in the pharmaceutical combination is dabrafenib.

[0057] In one embodiment, the MEK inhibitor contained in the pharmaceutical combination is a V600 mutant form of BR It is an inhibitor of AF.

[0058] In one embodiment, the MEK inhibitor contained in the pharmaceutical combination is V600E or V60 It is an inhibitor of the 0K mutant form of BRAF.

[0059] In one embodiment, the MEK inhibitor contained in the pharmaceutical combination is selumetinib (Astr aZeneca), binimetinib (Array Biopharma), PD-0325 901 (Pfizer), trametinib (Mekinist™), cobimetinib ( Exelixis, refametinib (Valeant), pimasertib (Santhe ra Pharmaceuticals), TAK-733 (Takeda) or WX -554 (UCB Pharma SA).

[0060] In one embodiment, the MEK inhibitor contained in the pharmaceutical combination is trametinib.

[0061] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and treatment with at least one anticancer drug selected from a BRAF inhibitor and a MEK inhibitor. an effective amount of the CDK inhibitor and at least one of the anti-cancer agents, wherein the CDK inhibitor and the at least one anti-cancer agent are administered simultaneously. This relates to combination medicines.

[0062] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and treatment with at least one anticancer drug selected from a BRAF inhibitor and a MEK inhibitor. and an effective amount of the CDK inhibitor and at least one of the anti-cancer agents, wherein the CDK inhibitor and the at least one anti-cancer agent are administered sequentially. The present invention relates to a combination drug.

[0063] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and treatment with at least one anticancer drug selected from a BRAF inhibitor or a MEK inhibitor. and an effective amount of the anticancer drug, wherein the anticancer drug is administered before the administration of the CDK inhibitor. do.

[0064] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and treatment with at least one anticancer drug selected from a BRAF inhibitor or a MEK inhibitor. and an effective amount of a CDK inhibitor, wherein the CDK inhibitor is administered before the administration of the anticancer drug. do.

[0065] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and treatment with at least one anticancer drug selected from a BRAF inhibitor or a MEK inhibitor. and an effective amount of the CDK inhibitor and the anti-cancer agent, wherein both the CDK inhibitor and the anti-cancer agent are administered once daily. This relates to pharmaceuticals.

[0066] In another embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma. a therapeutically effective amount of a CDK inhibitor selected from the group consisting of a compound of formula I, or a pharmaceutically acceptable salt thereof; and a therapeutic dose of at least one anticancer drug selected from a BRAF inhibitor or a MEK inhibitor. and a therapeutically effective amount of the CDK inhibitor, wherein the CDK inhibitor is administered once daily while the anticancer agent is administered twice daily. The present invention relates to a combination drug administered.

[0067] In yet another embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma. and a therapeutic agent for treating a CDK inhibitor selected from the group consisting of a compound of Formula I or a pharmaceutically acceptable salt thereof. and at least one anti-cancer drug selected from a BRAF inhibitor or a MEK inhibitor. and a therapeutically effective amount of an anti-cancer agent, wherein both the CDK inhibitor and the anti-cancer agent are administered twice daily. This relates to combination medicines.

[0068] In one aspect, the present invention provides a method for the treatment of melanoma, comprising administering to a patient of formula I (as described herein) or a pharmaceutically acceptable salt thereof. and at least one inhibitor selected from a BRAF inhibitor or a MEK inhibitor. and a therapeutically effective amount of a cancer drug to a subject in need thereof.

[0069] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a group consisting of a BRAF inhibitor and a BRAF inhibitor; and a therapeutically effective amount of at least one anticancer drug selected from MEK inhibitors. and administering the CDK inhibitor and the anticancer agent simultaneously to a subject. , relating to a method.

[0070] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a group consisting of a BRAF inhibitor and a BRAF inhibitor; and a therapeutically effective amount of at least one anticancer drug selected from MEK inhibitors. the CDK inhibitor and the anticancer agent are administered sequentially to a subject. This relates to a method.

[0071] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a group consisting of a BRAF inhibitor and a BRAF inhibitor; and a therapeutically effective amount of at least one anticancer drug selected from MEK inhibitors. the anticancer agent is administered to a subject having the disease before the administration of the CDK inhibitor. This relates to the method of

[0072] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a group consisting of a BRAF inhibitor and a BRAF inhibitor; and a therapeutically effective amount of at least one anticancer drug selected from MEK inhibitors. and administering the CDK inhibitor to a subject having a cancer, the CDK inhibitor being administered prior to administration of the anticancer agent. This relates to methods.

[0073] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a group consisting of a BRAF inhibitor and a BRAF inhibitor; and a therapeutically effective amount of at least one anticancer drug selected from MEK inhibitors. and administering to a subject, wherein the CDK inhibitor and the anti-cancer agent are both administered daily. The present invention relates to a method in which the compound is administered once.

[0074] In another embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or drug of formula I a therapeutically effective amount of a CDK inhibitor selected from the group consisting of a BRAF inhibitor and a BRAF inhibitor; and at least one therapeutically effective amount of a MEK inhibitor to a subject in need thereof. wherein the CDK inhibitor is administered once daily and the anticancer agent is administered twice daily. , relating to a method.

[0075] In yet another embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound of Formula I or or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CDK inhibitor selected from the group consisting of a BRAF inhibitor, a BRAF inhibitor, and a BRAF inhibitor. and at least one therapeutically effective amount of a therapeutically effective agent or MEK inhibitor to a subject in need thereof. wherein the CDK inhibitor and the anti-cancer agent are both administered twice daily. , relating to a method.

[0076] In one embodiment, the method of treating melanoma comprises administering to a subject a CDK inhibitor selected from a compound of Formula I and at least one anticancer agent selected from a BRAF inhibitor or a MEK inhibitor, This includes administering to a subject in need thereof in the dosage ranges described herein.

[0077] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a BRAF inhibitor, wherein the BRAF inhibitor and the CDK inhibitor are administered simultaneously.

[0078] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a BRAF inhibitor, wherein the BRAF inhibitor and the CDK inhibitor are administered sequentially.

[0079] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a BRAF inhibitor, wherein the BRAF inhibitor is administered in a manner similar to that of the CDK inhibitor. This relates to combination drugs that are administered before administration of the drug.

[0080] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a BRAF inhibitor, wherein the CDK inhibitor is The present invention relates to a combination drug administered prior to administration of the other drug.

[0081] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a BRAF inhibitor, wherein the CDK inhibitor and the BRAF inhibitor are both administered once daily.

[0082] In another embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma. a therapeutically effective amount of a CDK inhibitor selected from the group consisting of a compound of formula I, or a pharmaceutically acceptable salt thereof; and a therapeutically effective amount of a BRAF inhibitor, wherein the CDK inhibitor is administered once daily; The present invention relates to a combination pharmaceutical, wherein the BRAF inhibitor is administered twice daily.

[0083] In yet another embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma. and a therapeutic agent for treating a CDK inhibitor selected from the group consisting of a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a BRAF inhibitor, The present invention relates to a combination drug, in which both F inhibitors are administered twice daily.

[0084] In one aspect, the present invention provides a method for the treatment of melanoma, comprising administering to a patient of formula I (as described herein) or a pharmaceutically acceptable salt thereof. and administering to a subject in need thereof a therapeutically effective amount of a BRAF inhibitor. Regarding the law.

[0085] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from among a therapeutically acceptable salt thereof and a therapeutically effective amount of a BRAF inhibitor; and administering to a subject in need thereof a therapeutically effective amount of said BRAF inhibitor and The CDK inhibitors are administered simultaneously.

[0086] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from among a therapeutically acceptable salt thereof and a therapeutically effective amount of a BRAF inhibitor; and administering to a subject in need thereof a therapeutically effective amount of said BRAF inhibitor and wherein the CDK inhibitors are administered sequentially.

[0087] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from among a therapeutically acceptable salt thereof and a therapeutically effective amount of a BRAF inhibitor; and administering to a subject in need thereof a therapeutically effective amount of the BRAF inhibitor to The present invention relates to a method in which the compound is administered prior to administration of the CDK inhibitor.

[0088] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from among a therapeutically acceptable salt thereof and a therapeutically effective amount of a BRAF inhibitor; and administering to a subject in need thereof a therapeutically effective amount of the CDK inhibitor. is administered prior to administration of a BRAF inhibitor.

[0089] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from among a therapeutically acceptable salt thereof and a therapeutically effective amount of a BRAF inhibitor; and administering to a subject in need thereof a therapeutically effective amount of said CDK inhibitor and said The method relates to a method in which both of the BRAF inhibitors are administered once daily.

[0090] In another embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or drug of formula I a therapeutically effective amount of a CDK inhibitor selected from a therapeutically acceptable salt thereof and a BRAF inhibitor; and a therapeutically effective amount of the CDK inhibitor administered to a subject in need thereof, wherein the CDK inhibitor is administered daily. The present invention relates to a method in which the BRAF inhibitor is administered twice daily, while the BRAF inhibitor is administered once daily.

[0091] In yet another embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound of Formula I or or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CDK inhibitor selected from the group consisting of a BRAF inhibitor, a BRAF inhibitor, and a BRAF inhibitor. and administering to a subject in need thereof a therapeutically effective amount of a CDK inhibitor. and wherein said BRAF inhibitors are both administered twice daily.

[0092] In one embodiment, the method of treating melanoma comprises combining a BRAF inhibitor with a compound or drug of Formula I and a CDK inhibitor selected from the group consisting of a medicament for treating a pulmonary arthritis and a physiologically acceptable salt thereof, This includes administering to a subject in need thereof a range of amounts.

[0093] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a MEK inhibitor, wherein the MEK inhibitor and the CDK inhibitor are administered simultaneously. It relates to a combination drug administered at the same time.

[0094] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a MEK inhibitor, wherein the MEK inhibitor and the CDK inhibitor are administered sequentially. The present invention relates to a combination drug administered to a patient.

[0095] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a MEK inhibitor, wherein the MEK inhibitor is administered after administration of the CDK inhibitor. The present invention relates to a combination drug administered before the administration of the present invention.

[0096] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a MEK inhibitor, wherein the CDK inhibitor is administered after administration of the MEK inhibitor. The present invention relates to a combination drug administered before the administration of the present invention.

[0097] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a MEK inhibitor, wherein the CDK inhibitor and the MEK inhibitor are both Both drugs are administered once a day.

[0098] In another embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma. a therapeutically effective amount of a CDK inhibitor selected from the group consisting of a compound of formula I, or a pharmaceutically acceptable salt thereof; and a therapeutically effective amount of a MEK inhibitor, wherein the CDK inhibitor is administered once daily; wherein the MEK inhibitor is administered twice a day.

[0099] In yet another embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma. and a therapeutic agent for treating a CDK inhibitor selected from the group consisting of a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a CDK inhibitor and a therapeutically effective amount of a MEK inhibitor, The present invention relates to a combination drug in which both inhibitors are administered twice daily.

[0100] In one aspect, the present invention provides a method for the treatment of melanoma, comprising administering to a patient of formula I (as described herein) or a pharmaceutically acceptable salt thereof. and administering a therapeutically effective amount of a MEK inhibitor to a subject in need thereof. Regarding.

[0101] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a therapeutically acceptable salt thereof and a therapeutically effective amount of a MEK inhibitor; and administering to a subject in need thereof an effective amount of said MEK inhibitor and said The method relates to a method in which a CDK inhibitor is administered simultaneously.

[0102] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a therapeutically acceptable salt thereof and a therapeutically effective amount of a MEK inhibitor; and administering to a subject in need thereof an effective amount of said MEK inhibitor and said The CDK inhibitors are administered sequentially.

[0103] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a therapeutically acceptable salt thereof and a therapeutically effective amount of a MEK inhibitor; and administering to a subject in need thereof an effective amount of the MEK inhibitor to the subject, wherein the MEK inhibitor inhibits the CD The present invention relates to a method in which the compound is administered prior to the administration of a K inhibitor.

[0104] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a therapeutically acceptable salt thereof and a therapeutically effective amount of a MEK inhibitor; and administering to a subject in need thereof an effective amount of the CDK inhibitor to the ME. The present invention relates to a method in which the compound is administered prior to the administration of a K inhibitor.

[0105] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from a therapeutically acceptable salt thereof and a therapeutically effective amount of a MEK inhibitor; and administering to a subject in need thereof an effective amount of said CDK inhibitor and said and wherein both MEK inhibitors are administered once daily.

[0106] In another embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or drug of formula I a therapeutically effective amount of a CDK inhibitor selected from a therapeutically acceptable salt thereof and a therapeutically effective amount of a MEK inhibitor; to a subject in need thereof, wherein the CDK inhibitor is administered once daily in a therapeutically effective amount. wherein the MEK inhibitor is administered twice daily.

[0107] In yet another embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound of Formula I or or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CDK inhibitor selected from the group consisting of a medicament for treating rheumatoid arthritis and a medicament for treating rheumatoid arthritis; and a therapeutically effective amount of said CDK inhibitor and and said MEK inhibitor are both administered twice daily.

[0108] In one embodiment, the method of treating melanoma comprises combining a MEK inhibitor with a compound or pharmaceutical agent of Formula I. and a CDK inhibitor selected from the group consisting of a medicament for the treatment of rheumatoid arthritis and a therapeutically acceptable salt thereof, at a dose as described herein. In a range of applications, this includes administering to a subject in need thereof.

[0109] In one embodiment, the present invention provides a pharmaceutical combination for use in the treatment of melanoma, comprising: A therapeutically effective amount of a CDK inhibitor selected from a compound of Formula I or a pharmaceutically acceptable salt thereof. and a therapeutically effective amount of a BRAF inhibitor and a therapeutically effective amount of a MEK inhibitor. Regarding.

[0110] In one embodiment, the present invention provides a method of treating melanoma, comprising administering to a subject a compound or pharmaceutical composition of formula I. a therapeutically effective amount of a CDK inhibitor selected from among a therapeutically acceptable salt thereof and a therapeutically effective amount of a BRAF inhibitor; and a therapeutically effective amount of a MEK inhibitor to a subject in need thereof. Well, regarding the method.

[0111] In one embodiment of the present invention, the melanoma being treated is non-refractory melanoma.

[0112] In another embodiment of the invention, the melanoma being treated is non-refractory BRAF mutant melanoma. do.

[0113] In yet another embodiment of the invention, the melanoma being treated is non-refractory BRAF V600 It is a mutant melanoma.

[0114] In a further embodiment of the invention, the melanoma being treated is non-refractory BRAF V600E or BRAF V600K mutant melanoma.

[0115] In one embodiment of the present invention, the melanoma being treated is recurrent or refractory melanoma.

[0116] In another embodiment of the invention, the melanoma being treated is a resistant BRAF mutant melanoma.

[0117] In yet another embodiment of the invention, the melanoma being treated is a resistant BRAF V600 mutation It is a type of melanoma.

[0118] In a further embodiment of the invention, the melanoma being treated is resistant BRAF V600E or is a BRAF V600K mutant melanoma.

[0119] In one embodiment of the present invention, the melanoma being treated is metastatic melanoma.

[0120] In another embodiment of the invention, the melanoma being treated is metastatic BRAF mutant melanoma. .

[0121] In yet another embodiment of the invention, the melanoma being treated is a metastatic BRAF V600 mutant. It is atypical melanoma.

[0122] In a further embodiment of the invention, the melanoma being treated is metastatic BRAF V600E or or BRAF V600K mutant melanoma.

[0123] According to the present invention, a CDK inhibitor (a compound of formula I), and / or a BRAF inhibitor and The administration of the anti-cancer agent selected from the group consisting of steroids and / or MEK inhibitors may be, without limitation, parenteral, oral, or intravenous. Any, including oral, sublingual, transdermal, topical, intranasal, aerosol, intraocular, intratracheal, or intrarectal The method can be carried out by any suitable route.

[0124] In one embodiment, the CDK inhibitor is administered intravenously to generate and maintain good blood levels of the inhibitor. and a BRAF inhibitor and / or a MEK inhibitor. The anticancer drug(s) used may be administered parenterally, intravenously, subcutaneously, intramuscularly, intravascularly, or by infusion. It can be administered by any route.

[0125] In another embodiment, the CDK inhibitor is administered parenterally, intravenously, subcutaneously, intramuscularly, intravascularly, or It can be administered by infusion, while BRAF inhibitors and / or MEK inhibitors The anti-cancer agent(s) selected from the anti-cancer agents can be administered orally.

[0126] In a further embodiment, a CDK inhibitor and a BRAF inhibitor and / or a MEK inhibitor and (a) an anticancer drug(s) selected from the group consisting of: It can be administered orally to

[0127] In yet a further embodiment, a CDK inhibitor of Formula I is combined with a BRAF inhibitor and / or M Both the anticancer drug(s) selected from EK inhibitors are administered intravenously, subcutaneously, intramuscularly, or intravenously. It is administered parenterally by intravenous or infusion route to achieve good blood concentrations. can be created and maintained.

[0128] In one aspect, the present invention provides a pharmaceutical composition for use in the treatment of melanoma, comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and a BRAF inhibitor. at least one anticancer drug selected from the group consisting of a cyclophosphamide, ... and a pharmaceutically acceptable carrier, excipient, or additive. For the production of coated tablets and hard gelatin capsules, it can be used Pharmaceutically active excipients include, but are not limited to, lactose, corn denat. Examples include gum arabic, magnesium, and glucose. For capsules and suppositories, the carriers that can be used are limited to these. Examples of liquids that are not suitable for use include fats, waxes, natural or hydrogenated oils. Suitable carriers for the production of injection solutions or for emulsions or syrups are, for example, water, physiological salts, Sodium chloride solution or alcohol, e.g., ethanol, propanol or glycerol glycerol, sugar solutions such as glucose or mannitol solutions, or other solutions as described It is a mixture of various solvents.

[0129] a CDK inhibitor (a compound of Formula I) and a BRAF inhibitor and / or a MEK inhibitor; The anti-cancer agent(s) selected may be any of the anti-cancer agents, individually or in combination, familiar to those skilled in the art. Using conventional pharmaceutical techniques, e.g., blending, granulation, dissolving or lyophilizing This allows it to be formulated into a pharmaceutical dosage form.

[0130] Generally, compositions intended for pharmaceutical use are prepared using methods known in the art for the manufacture of pharmaceutical compositions. Any method known in the art, e.g., Remington-The Science and Practice of Pharmacy (21st edition) (2005), Goodma n&Gilman's The Pharmacological Basis of Therapeutics (11th ed.) (2006) and Ansel's Pharma Maceutical Dosage Forms and Drug Deliver y Systems (9th Edition), and Solid-State Chemistry It can be prepared according to the method described in The Book of Drugs (2nd edition) (1999), etc.

[0131] The compositions described herein may be in a form suitable for oral administration, for example, a solid dosage form, e.g. liquid dosage forms, e.g., emulsions, solutions, suspensions; Forms suitable for parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), e.g. For example, sterile solutions, suspensions or emulsions; forms suitable for topical administration, for example, ointments, creams The formulation may be a gel or a lotion.

[0132] Compositions for oral administration include tablets, lozenges, aqueous or oily suspensions, granules, powders, etc. It may be in the form of a powder, wafer, emulsion, capsule, syrup or elixir. Orally administered compositions may contain one or more optional agents, such as sweeteners, e.g., flavoring agents, e.g., flavonoids, phenoxyethanol ... lucutose, aspartame, or saccharin; flavoring agents, e.g., peppermint , wintergreen oil or cherry; coloring agents; and preservatives. A more palatable formulation can be obtained. The membrane is also permeable to the compound (CDK) contained in the pharmaceutical combination according to the present invention. inhibitors, and / or BRAF inhibitors and / or MEK inhibitors Suitable for oral administration of anti-cancer drug(s). As ingredients, standard vehicles such as mannitol, lactose, starch, Starch, magnesium stearate, talc, sodium saccharin, cellulose Such vehicles are preferably pharmaceutical It is of the highest quality.

[0133] For ointments and creams, the active ingredients (CDK inhibitors and / or BRAF inhibitors) The anti-cancer agent(s) selected from the group consisting of an anti-cancer agent and / or a MEK inhibitor are Or formulated in a water-in-oil base.

[0134] For intramuscular, intraperitoneal, subcutaneous and intravenous use, the active ingredients (CDK inhibitors and and / or anticancer drugs selected from BRAF inhibitors and / or MEK inhibitors (multiple A sterile solution of 100mg / mL or 100mg / mL of acetaminophen is usually used, and the pH of the solution should be appropriately adjusted and buffered. be.

[0135] Furthermore, the compounds, i.e., CDK inhibitors contained in the pharmaceutical compositions, and / or Efficacy of anticancer drugs selected from BRAF inhibitors and / or MEK inhibitors The effect can be delayed or prolonged through appropriate formulations, e.g., slow dissolving Pellets of the compound can be prepared and incorporated into tablets or capsules. The technique involves creating pellets with several different dissolution rates and filling capsules with a mixture of the pellets. This can be improved by filling the tablet or capsule with Even parenteral preparations can be coated with a film that prevents dissolution during administration. The compound is placed in an oily or emulsified vehicle that allows it to disperse only slowly in serum. The substance can be made long acting by dissolving or suspending it.

[0136] a CDK inhibitor selected from the compounds of formula I, and a BRAF inhibitor, The effective dose of the anti-cancer agent(s) selected from the group consisting of a steroid agent and / or a MEK inhibitor is determined based on the disease. (melanoma) severity, severity of symptoms, age, sex, weight and sensitivity of patients, mode of administration The doses vary depending on the dose, time, interval and duration, nature and type of formulation, etc. In certain embodiments, the therapeutic agents contained in the pharmaceutical combination according to the present invention are those for which both agents are dependent. Those skilled in the art will appreciate that the therapeutic agent administered is administered within a time frame that is still active. Such a time frame can be determined by determining the decay life. As shown, the anticancer drugs contained in the pharmaceutical composition can be administered simultaneously or sequentially. Those skilled in the art will recognize that several variations are possible within the scope and spirit of the present invention. Let it be recognized that this is the case.

[0137] The dosage of the therapeutic agent to be administered should be selected to produce the desired effect. A suitable dosage of the inhibitor may be from about 5 mg to about 500 mg. The dose of DK inhibitors can cover a wide range depending on the severity of the melanoma to be treated. The dose to be administered daily can be selected to achieve the desired effect. Suitable doses may range from about 50 mg / day to 350 mg / day of the CDK inhibitor. If needed, higher or lower daily doses can also be administered.

[0138] In one embodiment, the BRAF inhibitor is administered at about 1 mg / day to about 2500 mg / day. This amount can be administered per day, per administration, or per cycle of treatment. The dose may be given in a single dose or in multiple doses.

[0139] In one embodiment, the MEK inhibitor is administered at about 0.01 mg / day to 2000 mg / day. This amount can be administered per day, or per administration, or per cycle of treatment. The compositions may be administered in a single dose or in multiple doses per person.

[0140] In one embodiment, the inhibitor is selected from a CDK inhibitor and a BRAF inhibitor and / or a MEK inhibitor. In another embodiment, both the CD and the anticancer drug(s) of choice are administered once daily. K inhibitors and anticancer drugs selected from BRAF inhibitors and / or MEK inhibitors (multiple In a further embodiment, both the CDK inhibitor and the agonist are administered once daily. while administering an anticancer agent selected from a BRAF inhibitor and / or a MEK inhibitor However, the compound(s) contained in the combination pharmaceutical according to the present invention are administered twice a day. The amount of each therapeutic agent used, when used in combination, should be less than that which produces the therapeutic effect when administered alone. For convenience, the total daily dose may be divided and administered in portions if desired. It can be administered in small doses over the course of a day.

[0141] The combinations provided herein are compatible with certain assay systems and several different administration strategies. The experimental details are described below in The data presented herein demonstrate that BRAF inhibitors or The MEK inhibitor is a CDK inhibitor selected from the group consisting of compounds of Formula I and pharmaceutically acceptable salts thereof: It has been clearly shown that when combined with inhibitors, they exhibit synergistic effects. When used in combination to treat tumors, the effects are similar to those seen when cells are treated with CDK inhibitors alone (i.e., i.e., a compound of Formula I or a pharmaceutically acceptable salt thereof alone) or a BRAF inhibitor Apoptosis in proliferating cells was significantly reduced compared with treatment with either HCl or MEK inhibitor alone. It has been clearly shown that it increases cytotoxicity.

[0142] In one aspect, the present invention provides a CDK inhibitor (a compound of Formula I or a pharmaceutically acceptable salt thereof): ) and at least one anticancer drug selected from a BRAF inhibitor or a MEK inhibitor. The pharmaceutical kit comprises a compound of formula I or a pharmaceutically acceptable salt thereof. a salt and at least one anticancer drug selected from a BRAF inhibitor or a MEK inhibitor; The kit may also include a container containing a compound of Formula I as a fixed dose formulation. or a pharmaceutically acceptable salt thereof, and a BRAF inhibitor or a MEK inhibitor. The kit may contain two or more separate containers for at least one anticancer drug to be administered. contains information on indications, usage, dosage, directions for administration, contraindications, precautions and warnings. Suitable containers that can be used include bottles, vials, etc. The pharmaceutical kit may be packaged in a blister pack, ampoule, syringe or blister pack. Acceptable buffers include water for injection, phosphate buffered saline, Ringer's solution, and dextrose solution. A further container containing a liquid may optionally be included.

[0143] The CDK inhibitor, i.e., the compound of formula I contained in the pharmaceutical combination of the present invention, is P CT Patent Publication WO2004004632 and PCT Patent Publication WO2007148158 It can be prepared according to the method disclosed in

[0144] A general method for the preparation of compounds of formula I or pharmaceutically acceptable salts thereof follows. Step: (a) in the presence of a Lewis acid catalyst, a compound of formula VIA

[0145] [ka] The intermediate compound was purified as an enantiomerically pure (-)-trans enantiomer. mer with acetic anhydride to give the compound of formula VIIA.

[0146] [ka] obtaining an acetylated compound of formula (I); (b) reacting the resolved acetylated compound of formula VIIA with a compound of formula Ar Acids of formula COOH or acid chlorides of formula ArCOCl or acid anhydrides of formula (ArCO)2O or are esters of formula ArCOOCH3, where Ar is as defined herein above in connection with compounds of formula I. (as defined above) to form a compound of formula VIIIA

[0147] [ka] obtaining a resolved compound of (c) treating the resolved compound of formula VIIIA with a base in a suitable solvent to obtain the compound of formula IX A

[0148] [ka] wherein Ar is as defined above, obtaining the corresponding resolved β-diketone compound of formula (I); (d) By treating the resolved β-diketone compound of formula IXA with an acid such as hydrochloric acid, , Formula XA

[0149] [ka] and obtaining the corresponding cyclized compound of formula (I) (e) by heating it with a dealkylating agent at a temperature in the range of 120 to 180°C. , by subjecting the compound of formula XA to dealkylation to give the (+)-t compound of formula I. and optionally converting the compound to its pharmaceutically acceptable salt. and converting it into a salt. Includes.

[0150] The Lewis acid catalyst utilized in step (a) above is BF3, Et2O, zinc chloride, ammonium chloride. It can be selected from aluminum and titanium chlorides.

[0151] The bases utilized in process step (b) are triethylamine, pyridine and DC C-DMAP (N,N'-dicyclohexylcarbodiimide and 4-dimethylaminomethylpropional) The compound may be selected from the group consisting of a cyclohexyl benzoate, ...

[0152] The rearrangement of the compound of formula VIIIA to the corresponding β-diketone compound of formula IXA can be carried out as described by Baker et al. Those skilled in the art will appreciate that this is known as the r-Venkataraman rearrangement (J Chem.Soc., 1933, p. 1381 and Curr.Sci., 1933 Year, Vol. 4, p. 214).

[0153] The base used in process step (c) is lithium hexamethyldisilazide, natrium Sodium hexamethyldisilazide, potassium hexamethyldisilazide, sodium hydride and and potassium hydride. A preferred base is lithium hexamethyldihydrogen. It's Shirazide.

[0154] used in process step (e) for the dealkylation of the compound of formula IXA Dealkylating agents include pyridine hydrochloride, boron tribromide, boron trifluoride etherate, and The preferred dealkylating agent can be selected from aluminum trichloride, pyridine hydrochloride, It's salt.

[0155] The preparation of the starting compound of formula VIA can be carried out by reacting 1-methyl-4-piperidone with 1,3,5-trimethicone. 1-methyl-4-(2,4,6-trimethyl-2,4-dimethyl-2,4-dihydroxybenzene by reaction with a solution of 2,4,6-trimethyl-2,4-dimethyl ... -trimethoxyphenyl)-1,2,3,6-tetrahydropyridine, Boron trifluoride diethyl etherate, sodium borohydride and tetrahydrofuran By reacting, 1-methyl-4-(2,4,6-trimethoxyphenyl)piperazine This involves producing lysin-3-ol. Conversion of triphenylpiperidin-3-ol to the compound of formula VIA is carried out by triethyl In the presence of oxygen nucleophiles such as diamine, pyridine, potassium carbonate, or sodium carbonate, p-Toluenesulfonyl chloride, methanesulfonyl chloride, triflic anhydride or pentachlorobenzene Treatment with a suitable reagent such as phosphorus chloride affords the compound, 1-methyl-4-(2,4,6- The hydroxyl group on the piperidine ring of (trimethoxyphenyl)piperidin-3-ol The aryl group is converted to a leaving group such as tosyl, mesyl, triflate or halide, followed by isopropanol in the presence of an oxygen nucleophile such as sodium acetate or potassium acetate. The ring contraction is carried out in an alcoholic solvent such as alcohol, ethanol, or propanol. include.

[0156] Representative compound used in pharmacological assays, Compound A (also known as voruciclib) is (+)-trans-2-(2-chloro-4-trifluorophenyl)-5,7-di Hydroxy-8-(2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chloro Men-4-one hydrochloride, as disclosed in published PCT publication WO2007148158 It was one of the compounds investigated.

[0157] Another representative compound used in the pharmacological assays, Compound B (also known as ribiciclib), (+)-trans-2-(2-chlorophenyl)-5,7-dihydroxybenzoate si-8-(2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chromene-4 hydroxybenzoate hydrochloride, a compound disclosed in published PCT publication WO2004004632. It was one of those things.

[0158] a CDK inhibitor and at least one inhibitor selected from a BRAF inhibitor or a MEK inhibitor The synergistic effect of the combination according to the invention, which comprises an anti-cancer agent, can be seen in the preferred embodiments thereof. These are provided by way of example only and are not intended to limit the invention. Note that this is unintentional.

[0159] The present invention embraces various alternative modifications and variations, except where expressly specified to the contrary. It should be understood that any sequence of steps may be assumed. or as used in the specification and claims unless otherwise indicated. All numbers expressing, for example, quantities of ingredients, are modified in all instances by the term "about." Therefore, unless specifically indicated to the contrary, The numerical parameters set forth in the following specification and appended claims are intended to define the present invention. The temperature may vary depending on the desired properties to be obtained.

[0160] Those skilled in the art will recognize that several variations are possible within the scope and spirit of the present invention. The present invention will now be described in more detail with reference to the following non-limiting examples. The following examples further illustrate the present invention but, of course, do not limit its scope. should never be construed as [Example]

[0161] The following abbreviations or terms are used herein: BF3: Boron trifluoride BF3.Et2O: Boron trifluoride diethyl etherate CaCl2: Calcium chloride CHCl3: Chloroform CDCl3: deuterated chloroform CO2: Carbon dioxide DCC: N,N'-dicyclohexylcarbodiimide DMAP: 4-dimethylaminopyridine DMF: N,N-dimethylformamide DMSO: dimethyl sulfoxide Et2O: Diethyl ether EtOAc: ethyl acetate g: grams h: time HCl: Hydrochloric acid IPA: Isopropyl alcohol KBr: Potassium bromide Kg: kilogram L: Liter MeOH: Methanol min:minutes mg: milligram mL: milliliter μL: microliter μM: micromolar mmol: millimolar mol: mole NaCl: Sodium chloride Na2CO3: Sodium carbonate NaHCO3: Sodium bicarbonate Na2SO4: Sodium sulfate n-BuLi: n-butyllithium PEL:Piramal Enterprises Limited ℃: Celsius THF: tetrahydrofuran

[0162] example Preparation of Compound A (voruciclib) and Compound B (ribiciclib), representative compounds of Formula 1 The following are provided herein by way of reference:

[0163] Reference Example 1: (a)(+)-trans-2-(2-chloro-4-trifluoromethylphenyl)-8 -(2-hydroxymethyl-1-methylpyrrolidin-3-yl)-5,7-dimethoxy- Preparation of chromen-4-one n-BuLi (15% solution in hexane, 2.2 mL) kept at 0 °C under nitrogen atmosphere 5 mmol) in THF (10 mL), to a solution of hexamethyldisilazane (1.08 mL , 5.1 mmol) was added dropwise and stirred for 15 min. , (+)-trans-2-chloro-4-trifluoromethylbenzoic acid 2-(2-acetate (1-methyl-pyrrolidin-3-yl)-6-acetyl-3,5-dimethoxy A solution of the phenyl ester (1.44 g, 2.5 mmol) in THF (10 mL) was added dropwise. After the addition, the reaction was allowed to warm to room temperature and stirred for 2.5 h. The reaction mixture was acidified with diluted HCl. The aqueous layer was acidified and basified to pH 8-9 with 10% sodium bicarbonate. The organic layer was washed with water (25 mL), brine (25 mL), and The organic layer was concentrated under reduced pressure and dried under vacuum. , 3-{3-[3-(2-chloro-4-trifluoromethyl-phenyl)-acetate as an oil 3-oxo-propionyl]-2-hydroxy-4,6-dimethoxy-phenyl}-1- Methyl-pyrrolidin-2-ylmethyl ester was produced (1.3 g, 90.2%). The ester was dissolved in concentrated HCl (10 mL) and stirred for 3 h to allow cyclization to occur. Finally, the reaction mixture was basified with solid NaHCO3 to pH 8-9. The mixture was extracted with ethanol (25×3 mL) and washed with water (25 mL) and brine (25 mL). The organic layer was dried over anhydrous Na2SO4, concentrated under reduced pressure, and dried in vacuo. Column chromatography with 3% methanol and 0.1% ammonia as the eluent The residue was purified by HPLC to give compound (+)-trans-2 as a yellow solid. -(2-chloro-4-trifluoromethylphenyl)-8-(2-hydroxymethyl-1 -methylpyrrolidin-3-yl)-5,7-dimethoxy-chromen-4-one was produced. . Yield: 0.56 g (48.2%); 1 H NMR (CDCl3, 300MHz): δ 7.95 (d, 1H), 7.78 (s, 1H), 7. 69 (d, 1H), 6.61 (s, 1H), 6.46 (s, 1H), 4.21 (m, 1H), 4.01 (s, 3H), 3.93 (s, 3H) , 3.71 (dd, 1H), 3.41 (d, 1H), 3.26 (m, 1H), 2.84 (m, 1H), 2.70 (m, 1H), 2.44 (s , 3H), 2.10 (m, 2H); MS (ES+): m / z 497 (M+1).

[0164] (b) (+)-trans-2-(2-chloro-4-trifluoromethyl-phenyl)- 5,7-Dihydroxy-8-(2-hydroxymethyl-1-methylpyrrolidine-3-yl) Preparation of (4-methyl)-chromen-4-one The compound obtained in part (a) (0.25 g, 0.5 mmol), pyridine hydrochloride (0.2 A mixture of 5 g (2.16 mmol) and a catalytic amount of quinoline was heated at 180°C for 2.5 h. The reaction mixture was diluted with methanol (25 mL) and salted to pH 10 with solid Na2CO3. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated to give 0.1% Column chromatography using 4.5% methanol in ammonia and chloroform as eluents. The residue was purified by HPLC to give compound (+)-tr as a yellow solid. ans-2-(2-chloro-4-trifluoromethylphenyl)-5,7-dihydroxy -8-(2-hydroxy-methyl-1-methylpyrrolidin-3-yl)-chromene-4- Generated an on. Yield: 0.15 g (63.7%); 1 H NMR (CDCl3, 300MHz): δ 7.99 (m, 2H), 7.83 (d, 1H), 6 .65 (s, 1H), 6.41 (s, 1H), 4.24 (m, 1H), 3.90 (m, 2H), 3.70 (m, 1H), 3.60 (m, 1H) ), 3.41 (m, 1H), 2.99 (s, 3H), 2.54 (m, 1H), 2.28 (m, 1H); MS (ES+): m / z 470 (M+ 1).

[0165] (c) (+)-trans-2-(2-chloro-4-trifluoromethylphenyl)-5 ,7-dihydroxy-8-(2-hydroxymethyl-1-methylpyrrolidin-3-yl Preparation of )-chromen-4-one hydrochloride (compound A or voruciclib) The compound obtained in (b) (0.1 g, 0.2 mmol) was suspended in methanol (2 mL). Compound (+)- was obtained by treating with ethereal HCl and evaporating the organic solvent. trans-2-(2-chloro-4-trifluoromethyl-phenyl)-5,7-dihydriodide Roxy-8-(2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chromene -4-one hydrochloride was produced. Yield: 0.1 g (92.8%); 1 H NMR (CDCl3, 300MHz): δ 8.02 (d, 2H), 7.83 (d, 1H), 6.6 4 (s, 1H), 6.41 (s, 1H), 4.23 (m, 1H), 3.73 (m, 2H), 3.68 (m, 1H), 3.51 (m, 1H), 3.39 (m, 1H), 2.99 (s, 3H), 2.54 (m, 1H), 2.31 (m, 1H).

[0166] Reference Example 2: (a) (+)-trans-2-(2-chlorophenyl)-8-(2-hydroxymethyl) Preparation of (1-methylpyrrolidin-3-yl)-5,7-dimethoxy-chromen-4-one With stirring under a nitrogen atmosphere, sodium hydride (50%, 0.54 g, 11.25 m mol) to (-)-trans-1-[2-hydroxy-3-(2-hydroxymethyl -1-methylpyrrolidin-3-yl)-4,6-dimethoxyphenyl)-ethanone (0. 7 g, 2.2 mmol) in dry DMF (15 mL) was added portionwise at 0°C. After 0 min, methyl 2-chlorobenzoate (1.15 g, 6.75 mmol) was added. The reaction mixture was stirred at 25°C for 2 h. Methanol was added carefully below 20°C. The mixture was poured onto crushed ice (300 g) and acidified with 1:1 HCl (pH 2). Extraction was performed using EtOAc (2 x 100 mL). The aqueous layer was basified using and extracted with CHCl3 (3 x 200 mL). The mixture was dried (anhydrous Na2SO4) and concentrated. To the residue was added concentrated HCl (25 mL) and the mixture was heated at room temperature. The reaction mixture was poured onto crushed ice (300 g) and added saturated Na2CO3. The mixture was made basic using aqueous sodium hydroxide. CHCl3 (3 x 200 mL) was used to extract the mixture. The organic extract was washed with water, dried (anhydrous Na2SO4), and concentrated to give The compound, (+)-trans-2-(2-chloro-phenyl)-8-(2-hydroxymethyl) (1-methyl-pyrrolidin-3-yl)-5,7-dimethoxy-chromen-4-one obtained. Yield: 0.67 g (64 %); mp: 91 - 93℃; [α] D 25 = + 5.8°(c = 0.7, methanol); IR ( KBr): 3431, 1648, 1598, 1571 cm -1 ; 1 H NMR (CDCl3, 300MHz): δ 7.70 (dd, 1H), 7. 52 (m, 1H), 7.45 (m, 2H), 6.50 (s, 1H), 6.44 (s, 1H), 4.17 (m, 1H), 4.00 (s, 3H) , 3.97 (s, 3H), 3.64 (dd, 1H), 3.40 (d, 1H), 3.15 (m, 1H), 2.74 (d, 1H), 2.52 (m , 1H), 2.32 (s, 3H), 2.00 (m, 2H); MS (ES+): m / z 430 (M+1).

[0167] (b) (+)-trans-2-(2-chlorophenyl)-5,7-dihydroxy-8- (2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chromen-4-one preparation Molten pyridine hydrochloride (4.1 g, 35.6 mmol) of the compound obtained in part (a) (0.4 g, 0.9 mmol) was added and heated at 180° C. for 1.5 h. Cool to °C, dilute with MeOH (10 mL), and basify to pH 10 using Na2CO3 The mixture was filtered and the organic layer was concentrated. The residue was suspended in water (5 mL) and stirred for 30 min. The compound, (+)-trans-2-(2- chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1-methyl -pyrrolidin-3-yl)-chromen-4-one was obtained. Yield: 0.25 g (70%); IR (KBr): 3422, 3135, 1664, 1623, 1559 cm -1 ; 1 H NMR (CDCl , 300MHz): δ 7.56 (d, 1H), 7.36 (m, 3H), 6.36 (s, 1H), 6.20 (s, 1H), 4.02 (m, 1 H), 3.70 (m, 2H), 3.15 (m, 2H), 2.88 (m, 1H), 2.58 (s, 3H), 2.35 (m, 1H), 1 .88 (m, 1H); MS (ES+): m / z 402 (M+1); Analysis: C 21 H 20 ClNO5C, 62.24 (62.71); H, 5.07 (4 .97); N, 3.60 (3.48); Cl, ​​9.01 (8.83).

[0168] (c) (+)-trans-2-(2-chlorophenyl)-5,7-dihydroxy-8- (2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chromen-4-one salt Preparation of the Acid Salt (Compound B or Ribiciclib) The compound obtained in part (b) (0.2 g, 0.48 mmol) was suspended in IPA (5 mL). The mixture was stirred for 2 hours, and 3.5% HCl (25 mL) was added. The suspension was heated to give a clear solution. The solution was cooled and the solid was filtered to obtain the compound, (+)-trans-2-(2- chlorophenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1-methyl- Pyrrolidin-3-yl)-chromen-4-one hydrochloride was obtained. Yield: 0.21 g (97 %); mp: 188 - 192℃; [α] D 25 = +21.3°(c = 0.2, methanol); 1 H NMR (CD3OD, 300MHz): δ 7.80 (d, 1H), 7.60 (m, 3H), 6.53 (s, 1H), 6.37 (s, 1H) , 4.23 (m, 1H), 3.89 (m, 2H), 3.63 (m, 1H), 3.59 (dd, 1H), 3.38 (m, 1H), 2.90 (s , 3H), 2.45 (m, 1H), 2.35 (m, 1H); MS (ES+): m / z 402 (M +1) (free base).

[0169] Biological Data: Pharmacological assays: [Example]

[0170] I. Compound A (CDK inhibitor, bolus thrombin) in BRAF V600E-mutated melanoma cell lines a combination of ciclib (also called ciclib) and vemurafenib (a BRAF V600E inhibitor) In vitro studies, including the use of the purpose: Compound A (CDK inhibitor, also known as voruciclib) and vemurafenib (BRAF Combination of BRAF V600E inhibitors and BRAF V600E inhibitors in cellular responses in BRAF V600E-mutated melanoma cell lines The effects on cell cycle and apoptosis are studied.

[0171] material and method: cell line G361 and SK-M obtained from ATCC (American Type Culture Collection), USA The EL3 melanoma cell line was used in this study. G361 is a vemurafenib-sensitive cell line. SK-MEL3 is a vemurafenib-resistant cell line. Both cell lines express BRAF V. It has a 600E mutation.

[0172] A. Analysis of cell cycle distribution using flow cytometry: G361 / SK-MEL3 melanoma cells were grown at 25 mm 3 The cells were seeded into tissue culture flasks. After 24 h, G361 cells were treated for 5 days with: i) Compound A (1 μM); ii) Bem rafenib (1 μM); and iii) Compound A (1 μM) and vemurafenib (1 μM). Together with.

[0173] For SK-MEL3 melanoma cells, cells were treated for 5 days with: i) Compound A (1 μg) ii) vemurafenib (10 μM); and iii) compound A (1 μM) with vemurafenib together with phenytoin (10 μM).

[0174] Control cells were left untreated for 5 days. Both detached and adherent cells were cultured at the end of the 5-day period. The cells were collected in approximately 5 mL of liquid by centrifugation at 1000 rpm for 10 min. The cells were washed twice with phosphate-buffered saline (PBS). The cells were resuspended in 500 μl of PBS. The cells were then fixed in 500 μl of ice-cold 70% ethanol at room temperature for 30 min. The mixture was incubated and spun at 1000 rpm for 10 minutes. The cell pellet was then diluted with 1 mL of Chilled 70% ethanol was added and stored below 0°C until further analysis. The cells were washed twice with PBS to remove the fixative and resuspended in 250 μl of PBS. 0.5 μl of propidium iodide (1 mg / mL in PBS) and 12.5 μl of RNAs e A (1 mg / ml) was added. After 30 min of incubation at 37°C, Cells were analyzed using cytometry.

[0175] These studies were performed using a flow cytometer (Becton Digital) according to the manufacturer's recommendations. A Kinson FACS Calibur (Mitsubishi UFJ, USA) was used. The wavelength was set at 488 nm. An argon ion laser was used as the excitation source. Cells were cultured at G1, S and G2 / M stages of the cell cycle as defined by the level of red fluorescence. Cells showing DNA content less than 2n were designated as sub-G1 cells. The number of cells in each cell cycle compartment was expressed as a percentage of the total number of cells present.

[0176] B. Annexin V-FITC staining (for detecting early apoptosis) AnnexinV-FITC is a sensitive probe for identifying apoptotic cells. During early apoptosis, the membrane phospholipid phosphotidylserine (PS) is transported to the inner nucleus of the plasma membrane. A translocates from the cytosol to the outer leaflet, exposing PS to the external cellular environment. Annexin V has a high affinity for PS and binds to cells with exposed PS. It is a 35-36 kDa calcium-dependent phospholipid-binding protein.

[0177] Propidium iodide enters cells through leaky membranes and therefore slows apoptosis. It has been used in conjunction with fluorescein isothiocyanate (FITC) for the detection of It is a polar pigment.

[0178] Melanoma cells G361 were cultured at 25 mm 3 After 24 h, the cells were seeded into tissue culture flasks. Treatment for 24 h was with: i) Compound A (1 μM); ii) Vemurafenib (1 μM); and and iii) Compound A (1 μM) and vemurafenib (1 μM) together. Control cells were not The treatment was allowed to continue for 24 hours. The medium containing the mobile cells was collected and analyzed at different time points. After harvesting with trypsin, the cells were pooled with the adherent cells and centrifuged at 1000 rpm for 10 minutes. The cells were washed twice with cold PBS while being separated. 6 Cells at a concentration of cells / ml The pellet was resuspended in 1x binding buffer (10 mM HEPES pH 7.4, 140 mM NaCl , 2.5 mM CaCl2). 5 cells) to A The cells were stained with annexin V-FITC and propidium iodide. Incubate at room temperature (25-30°C) for 15 minutes and analyze the samples by flow cytometry. did.

[0179] result: The results of these tests are shown in Figures 1-3.

[0180] Conclusion: Figure 1 shows that G361 cells treated with vemurafenib showed significantly higher cell proliferation rates than control cells (72.36%). They pointed out that the G1 arrest rate was 91.94%. As can be seen from the results, cells treated with Compound A and vemurafenib alone showed 9% IL-1 expression, respectively. Compound A and Bemuraf showed only 0.48% and 2.3% apoptosis, respectively. In cells treated with the combination of α- and β-lactamase, apoptosis occurred in 53.67% of the cells. This indicates that the combination is synergistic.

[0181] In Figure 2, the upper right quadrant shows G361 cells in early apoptosis. These cells then undergo apoptosis, leading to cell death. Cells treated with the combination of Compound A and vemurafenib were significantly different from those treated with Compound A and vemurafenib alone, respectively. 27% of cells were in early apoptosis (a) compared with 8% and 20% for the transplanted cells. This data indicates that this combination has a synergistic effect. It is pointed out that this shows

[0182] In Figure 3, SK-MEL3 cells treated with vemurafenib alone showed significantly higher cell proliferation rates than control cells (67.3 8%), a significant G1 arrest was observed in 76.78% of cases. Thus, apoptosis was 29.75% in cells treated with the combination of Compound A and vemurafenib. %, while cells treated with Compound A and vemurafenib alone showed 1% The apoptosis rates were only 9.63% and 15.12%, respectively, demonstrating that this combination is synergistic. He points out that:

[0183] The results illustrated in Figures 1-3 and described above demonstrate that Compound A and vemurafenib The combination of vemurafenib and BRAF V600E-mutated melanoma is The combination was synergistic in cell lines and was superior to Compound A and vemurafenib used alone. It was clearly demonstrated that the combination induces more apoptosis than the single-agent combination. He stood up.

[0184] II. Compound A and Vemurafenib (BRAF) in Human BRAF-Mutated Melanoma Cells In vitro double combination test with V600E inhibitor Objective: The purpose of this study was to evaluate the efficacy of Compound A (a CDK inhibitor) in BRAF-mutated melanoma cells. The purpose of this study was to evaluate the efficacy of the combination of cefotaxime and vemurafenib (a BRAF V600E inhibitor). Ta.

[0185] A. Material Test compounds: Compound A (prepared by PEL'S Lab); Vemurafenib (Nanjing Chemlin Chemical Industry Co., Ltd, China) Vehicle: DMSO (Sigma-Aldrich-Chemie Gmbh, Germany) Dosing preparation: Compound A and vemurafenib were weighed and dissolved in the required amount of DMSO. , to obtain the required stock solution. Test system: The test system is G361, A375 and MDAMB-435S (BR These included the AF V600E mutant cell lines. n Type Culture Collection, USA.

[0186] B. Method Cytotoxicity studies using various combinations of Compound A and vemurafenib were performed on live CCK8 cells. This was done using a dehydrogenase assay.

[0187] i) Cell Counting Kit-8 (CCK8) Live Cell Dehydrogenase Assay Human BRAF V600E-mutated melanoma cancer cell lines, G361, A375, and MD AMB-435S was cultured in 199 μl of RPMI 1640 medium in a 96-well plate. Plate at a density of 3000 cells / well and incubate overnight to allow cells to attach. The cells were then treated with each test compound. There were 10 groups in total; i) 1 μM vemurafenib alone for 48 h; ii) 0.3 μM / 0.1 μM Compound A alone for 48 h; iii) 1 μM / 0.3 μM Compound A alone for 48 h; iv) 3 μM / 1 μM Compound A alone for 48 h; v) 10 μM / 3 μM Compound A alone for 48 h; vi) 0.3 μM / 0.1 μM Compound A in combination with 1 μM vemurafenib for 48 h; vii) 1 μM / 0.3 μM Compound A in combination with 1 μM vemurafenib for 48 h; viii) 3 μM / 1 μM Compound A in combination with 1 μM vemurafenib for 48 h; ix) 10 μM / 3 μM Compound A in combination with 1 μM vemurafenib for 48 h; x) Control wells were treated with DMSO vehicle for 48 h.

[0188] Plates were incubated at 37° ± 1°C in a humidified 5% CO2 incubator. Vemurafenib was used at a concentration of 1.0 μM in all three different cell lines. Compound A inhibited the growth of A375 melanoma cells at 0.3 μM, 1 μM, 3 μM, and 10 μM. For G361 and MDAMB435S melanoma cells, use concentrations of 0.1 μM and 0 μM. At the end of the incubation period, the following concentrations were used: 0.3 μM, 1.0 μM, and 3.0 μM. Plates were then assayed using the CCK8 cytotoxicity assay protocol. and combination index using Compusyn software by Talalay (4). Synergy was determined by calculating the CI (Comparison Indicator). CI<1 indicates synergistic activity, and CI= 1 is additive and CI>1 is antagonistic.

[0189] Statistical analysis: Statistical analysis was performed using the Student's t-test, and a p-value <0.05 was considered significant. Data are expressed as mean ± standard error of the mean (SEM). Obtained from at least two independent experiments performed in replicate.

[0190] ii) Cytotoxicity Assay Protocol Logarithmically growing cells are divided into 3x10 3 Plate at a density of 16–20 cells / well. The concentration of both Compound A and Vemurafenib was varied for 48 hours. After 48 hours, cytotoxicity was assessed using CCK-8 reagent (Dojindo Mol ecular Technologies, Inc., Maryland, and Japa According to the manufacturer's instructions, 5 μL / well of CCK-8 reagent was added and the plate was The rate was incubated for 2 h. The wavelength was corrected from 450 nm to 650 nm and the control and normalized on a Tecan Sapphire multi-fluorescence microplate reader. Toxicity was determined by measuring the absorbance of the ozone layer. All experiments were performed in triplicate. .

[0191] Treatment Schedule for Compound A and Vemurafenib: Tables 1 and 2 show the results of the ELISA in A375 cells, G361 cells, and MDA-MB435S cells. 1 shows the respective treatment schedules for Compound A and vemurafenib.

[0192] [Table 1]

[0193] [Table 2]

[0194] result: Cellular and Cellular Studies of Compound A and Vemurafenib and Their Combination in Different Cell Lines IC of cytotoxicity 50 The values ​​are presented in Table 3. Figures 4-6 and Tables 4-6 show the results for various melanoma cells. 1 shows the effect of the combination of Compound A and vemurafenib in cell lines.

[0195] [Table 3]

[0196] In Figures 4 to 6, the X axis represents the concentration of Compound A and Vemurafenib alone and in combination. The left Y-axis shows the cytotoxicity (%) and the right Y-axis shows the concentration of the compound when combined. The index value is shown.

[0197] In Figure 4, cells treated with 1 μM vemurafenib showed 23% inhibition, whereas Cells treated with 3 μM Compound A showed 46% inhibition of cells. When cells were treated with vemurafenib in combination with Compound A at the concentrations below The compatibility index (CI) was 0.70, and a synergistic effect was observed with 83% inhibition of cells. The synergy data presented are the average of two independent experiments, each performed in triplicate. means the value.

[0198] [Table 4]

[0199] In Figure 5, cells treated with 1 μM vemurafenib showed 30% inhibition, whereas Therefore, cells treated with 0.3 μM Compound A showed only 2% inhibition of cell proliferation. At this suboptimal concentration, cells were treated with vemurafenib in combination with Compound A. The combination index value was 0.77, demonstrating a remarkable synergistic effect of 52% inhibition of cells. Cells were treated with compound A (3 μM) in combination with vemurafenib (1 μM). When placed together, they showed 91% inhibition of cells with a combination index of 0.9. Mean values ​​are for two independent experiments, each performed in triplicate.

[0200] [Table 5]

[0201] In Figure 6, cells treated with 1 μM vemurafenib showed 31% inhibition of cells. In contrast, cells treated with 1 μM of Compound A showed 32% inhibition of the cells. At this point, when cells were treated with vemurafenib in combination with Compound A, the combination A synergistic effect was observed with a cell count of 0.74, resulting in 90% inhibition of cells. The synergistic data presented are Mean values ​​are for two independent experiments, each performed in triplicate.

[0202] [Table 6]

[0203] Conclusion: The combination of vemurafenib and Compound A showed significant efficacy in BRAF-mutated melanoma cells. A synergistic effect was observed.

[0204] References: 1.Smalley KS, Lioni M, Palma MD, Xiao M, Des ai B, Egyhazi S, Hansson J, Wu H, King AJ, Va. n Belle P, Elder DE, Flaherty KT, Herlyn M, Nathanson KL;「Increased cyclin D1 expres sion can mediate BRAF inhibitor resistan ce in BRAF V600E-mutated melanomas”;Mol. Cancer Ther., 2008, Vol. 7, pp. 2876-2883. 2.Smalley KSM and Flaherty KT; ng BRAF / MEK inhibitors into combination therapy for Melanoma”;British Journal of Cancer, 2009, vol. 100, pp. 431-435. 3. Dhomen N, Marais R; “BRAF signaling and "targeted therapies in melanoma" Hematol.O ncol.Clin.North Am., 2009, Volume 23, No. 3, 529~45 page. 4.Ting-ChaoChou; “Theoretical basis, expert imental design, and computerized simulati on of synergism and antagonismin drug co "Combination studies," Pharmacol.Rev., 2006, Vol. Volume 58, pages 621-81. [Example]

[0205] A. Human BRAF-V600E mutated melanoma cell line (A375) and its vemurafe Compound A (CDK inhibitor, also known as voruciclib) in nib-resistant derivatives (A375R) and BRAF inhibitors (vemurafenib or dabrafenib) or MEK inhibitors In vitro combination study with one anticancer drug selected from the group consisting of trametinib

[0206] the purpose: The purpose of this study was to investigate the effect of CDK inhibitor (voruciclib) on BRAF-mutated melanoma cells. ) and a BRAF inhibitor (vemurafenib or dabrafenib) or a MEK inhibitor ( The objective of this study was to evaluate the efficacy of the combination of rituximab (rituximab) and rametinib.

[0207] B. Material: Test compounds: Compound A (prepared by PEL'S Lab.); Vemurafenib (Selle ckchem USA, S1267); dabrafenib (Selleckchem US A, S2807) and trametinib (Selleckchem USA, S2673) Vehicle: DMSO (Sigma-Aldrich-Chemie Gmbh, Germany) Dosing preparation: Test compound was weighed and dissolved in the required amount of DMSO to obtain the required stock solution. A solution was obtained. Test system: The test system included the A375 (BRAF V600E mutant) cell line, which is a CC (American Type Culture Collection (USA) Cell lineage collection), USA and A375R cell line (vemurafenib-resistant-PE (Developed at L'S Lab.)

[0208] C. Method: All test compounds were evaluated as single agents using the CCK8 live cell dehydrogenase assay. Cytotoxicity studies were performed using both the 2000 and 2001 hydroxybenzoates in combination.

[0209] Cell Counting Kit-8 (CCK8) Viable Cell Dehydrogenase Assay: Logarithmically growing human BRAF V600E mutated melanoma cells were cultured at 1500 cells / well. 30 μL of the dilution was placed in a 384-well plate (Corning, USA) at a density of 100 μL. Beck's Modified Eagle Medium (DMEM) was incubated with 100 mL of PBS in a 5% CO2-free atmosphere using a Tecan automated platform (Fr Sow seeds using the eedomEvo Liquid Handling System and let them sow for approximately 12-16 hours. The cells were then incubated with different doses of Compound A and anti- Cancer drugs (vemurafenib or dabrafenib or trametinib) as monotherapy (used individually) and in combination for 48 h treatment. The results were compared for each combination (e.g., the combination of Compound A and vemurafenib). Treatment ratios of test compounds when used individually and in combination in A375R cells The rates are shown in Tables 7A-7C (i.e., the combination of Compound A and vemurafenib; Compound A and da and the combination of Compound A and trametinib). The following doses were used: 1, 0.5, 0.25, 0.125, 0.06, 0.03, 0. 015; Vemurafenib (μM) 30, 15, 7.5, 3.75, 1.875, 0.9, 0.4, 0.2, 0.1, 0.05; Dabrafenib (μM) 3, 1.5, 0.75, 0 .37, 0.18, 0.09, 0.04, 0.02, 0.01, 0.005; Trametini Bu(μΜ)1, 0.5, 0.25, 0.125, 0.0625, 0.031, 0.015 , 0.007, 0.003, 0.001. As shown in Tables 7A-7C, the above doses The anticancer drug (vemurafenib, dabrafenib, or trametinib) is combined with compound A. The controls used were cells alone or cells plus vehicle (DMSO). )

[0210] Plates were incubated in a humidified 5% CO2 incubator at 37°±1°C for 48 h. After post-incubation, CCK8 reagent (Dojindo Molecular cular Technologies, Inc., Maryland, and Japan) Plates were assayed by adding 3 μl of CCK-8 reagent to the plate according to the manufacturer's instructions. The plate was incubated for 2 hours. At a normalized wavelength of 450 nm, Tecan Sapphire Multi-Fluorescent Micro-Plate Toxicity was determined by measuring absorbance on a detector. All experiments were performed in quadruplicate. did.

[0211] Chou Tal calculates the combination index (CI) (CI values ​​less than 1 indicate synergy) Calcusyn software (Biosoft, Ferguson) based on the array method , MO, USA) was used to quantify the potency of the combination.

[0212] Table 7A-7C: Compound A and anticancer drugs (Bam) in A375 and A375R cells Rafenib or dabrafenib or trametinib) as monotherapy (used individually) and the ratio of treatments when combined.

[0213] Compound A (denoted as A) and vemurafenib (denoted as V) alone or in combination The treatment ratios (shown as 'groups') are presented in Table 7A.

[0214] [Table 7]

[0215] Compound A (denoted as A) and dabrafenib (denoted as D) alone or in combination The treatment ratios (shown as 'groups') are presented in Table 7B.

[0216] [Table 8]

[0217] Compound A (denoted as A) and trametinib (denoted as T) alone or in combination The proportions of treatments (as indicated) are presented in Table 7C.

[0218] [Table 9]

[0219] result: The results of these tests are shown in Figures 7-13.

[0220] Conclusion: 1. Figures 7a and 7b show that the A375 cell line is highly sensitive to vemurafenib. It can be concluded that A375R is resistant to the virus, whereas A375R is resistant to the virus. Both A375R are equally sensitive to compound A (voruciclib). 2. From Figures 8a and 8b, the combination of compound A (voruciclib) and vemurafenib is A3 It can be concluded that the compound showed strong synergistic effects (CI<0.5) in 75 cell lines. 3. From Figures 9a and 9b, the combination of compound A (voruciclib) and vemurafenib is shown to be It can be concluded that the 375R cell line exhibited a strong synergistic effect (CI<0.5). . 4. From Figures 10a and 10b, the combination of Compound A (voruciclib) and Dabrafenib It can be concluded that the A375 cell line exhibits a strong synergistic effect (CI<0.5). . 5. From Figures 11a and 11b, the combination of Compound A (voruciclib) and Dabrafenib We conclude that the A375R cell line exhibits a strong synergistic effect (CI<0.5). do. 6. From Figures 12a and 12b, the combination of compound A (voruciclib) and trametinib is A It can be concluded that the compound showed a strong synergistic effect (CI<0.5) in the 375 cell line. 7. From Figures 13a and 13b, the combination of compound A (voruciclib) and trametinib is A It can be concluded that the 375R cell line exhibited a strong synergistic effect (CI<0.5). .

Claims

1. 1. A pharmaceutical combination for treating melanoma, comprising: 【Chemistry 1】 wherein Ar is 2-chloro-4-trifluoromethylphenyl. or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of at least one anticancer agent selected from vemurafenib, dabrafenib, and trametinib.

2. 2. The pharmaceutical combination of claim 1, wherein the CDK inhibitor is (+)-trans-2-(2-chloro-4-trifluoromethylphenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride.

3. The combination pharmaceutical of claim 1 or 2, wherein the anticancer drug is vemurafenib.

4. The combination drug according to claim 1 or 2, wherein the anticancer drug is dabrafenib.

5. A combination pharmaceutical described in claim 1 or 2, wherein the anticancer drug is trametinib.

6. The pharmaceutical combination according to any one of claims 1 to 5, wherein the CDK inhibitor and the at least one anticancer agent are administered simultaneously to a subject in need thereof.

7. The pharmaceutical combination according to any one of claims 1 to 5, wherein the CDK inhibitor and the at least one anticancer agent are administered sequentially to a subject in need thereof.

8. The pharmaceutical combination according to any one of claims 1 to 7, wherein the melanoma is non-refractory melanoma.

9. 9. The pharmaceutical combination of claim 8, wherein the melanoma is non-refractory BRAF mutant melanoma.

10. 10. The pharmaceutical combination of claim 9, wherein the melanoma is non-refractory BRAF V600 mutant melanoma.

11. 11. The pharmaceutical combination of claim 10, wherein the melanoma is non-refractory BRAF V600E or BRAF V600K mutant melanoma.

12. The pharmaceutical combination according to any one of claims 1 to 7, wherein the melanoma is recurrent or refractory melanoma.

13. The pharmaceutical combination according to claim 12, wherein the melanoma is recurrent or refractory BRAF mutant melanoma.

14. The pharmaceutical combination of claim 13, wherein the melanoma is recurrent or refractory BRAF V600 mutant melanoma.

15. 15. The pharmaceutical combination of claim 14, wherein the melanoma is relapsed or refractory BRAF V600E melanoma or BRAF V600K mutant melanoma.

16. The pharmaceutical combination according to any one of claims 1 to 7, wherein the melanoma is metastatic melanoma.

17. 17. The pharmaceutical combination of claim 16, wherein the melanoma is metastatic BRAF mutant melanoma.

18. 18. The pharmaceutical combination of claim 17, wherein the melanoma is metastatic BRAF V600 mutant melanoma.

19. 19. The pharmaceutical combination of claim 18, wherein the melanoma is metastatic BRAF V600E melanoma or BRAF V600K mutant melanoma.

20. A pharmaceutical kit for treating melanoma, comprising: a compound of formula I 【Chemistry 2】 wherein Ar is 2-chloro-4-trifluoromethylphenyl. or a pharmaceutically acceptable salt thereof, and at least one anticancer drug selected from vemurafenib, dabrafenib, and trametinib.

21. The pharmaceutical kit according to claim 20, wherein the CDK inhibitor is (+)-trans-2-(2-chloro-4-trifluoromethylphenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1-methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride.

22. A pharmaceutical kit described in claim 20 or 21, wherein the anticancer drug is vemurafenib.

23. A pharmaceutical kit described in claim 20 or 21, wherein the anticancer drug is dabrafenib.

24. A pharmaceutical kit described in claim 20 or 21, wherein the anticancer drug is trametinib.

25. A pharmaceutical kit described in any one of claims 20 to 24, wherein the CDK inhibitor and the at least one anticancer agent are administered simultaneously to a subject in need thereof.

26. A pharmaceutical kit described in any one of claims 20 to 24, wherein the CDK inhibitor and the at least one anticancer agent are administered sequentially to a subject in need thereof.

27. ​​A pharmaceutical kit described in any one of claims 20 to 26, wherein the melanoma is non-refractory melanoma.

28. The pharmaceutical kit described in claim 27, wherein the melanoma is non-refractory BRAF mutant melanoma.

29. The pharmaceutical kit of claim 28, wherein the melanoma is non-refractory BRAF V600 mutant melanoma.

30. The pharmaceutical kit of claim 29, wherein the melanoma is non-refractory BRAF V600E or BRAF V600K mutant melanoma.

31. A pharmaceutical kit described in any one of claims 20 to 26, wherein the melanoma is recurrent or refractory melanoma.

32. The pharmaceutical kit described in claim 31, wherein the melanoma is recurrent or refractory BRAF mutant melanoma.

33. The pharmaceutical kit of claim 32, wherein the melanoma is recurrent or refractory BRAF V600 mutant melanoma.

34. The pharmaceutical kit of claim 33, wherein the melanoma is recurrent or refractory BRAF V600E melanoma or BRAF V600K mutant melanoma.

35. A pharmaceutical kit described in any one of claims 20 to 26, wherein the melanoma is metastatic melanoma.

36. The pharmaceutical kit described in claim 35, wherein the melanoma is metastatic BRAF mutant melanoma.

37. The pharmaceutical kit of claim 36, wherein the melanoma is metastatic BRAF V600 mutant melanoma.

38. The pharmaceutical kit of claim 37, wherein the melanoma is metastatic BRAF V600E melanoma or BRAF V600K mutant melanoma.

39. A pharmaceutical kit described in any one of claims 20 to 38, comprising a container containing the CDK inhibitor and the at least one anticancer agent as a fixed-dose formulation.

40. A pharmaceutical kit described in any one of claims 20 to 38, comprising a container containing the CDK inhibitor and one or more separate containers containing the at least one anticancer agent.

41. A pharmaceutical kit described in any one of claims 20 to 40, further comprising an insert containing information on indications, usage, dosage, administration instructions, contraindications, precautions and warnings.

42. The pharmaceutical kit of any one of claims 20 to 41, further comprising a container containing a pharmaceutically acceptable buffer, water for injection, phosphate-buffered saline, Ringer's solution, or dextrose solution.

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