Pharmaceutical composition for preventing or treating cancer

Abstract

An example of the present invention relates to a pharmaceutical composition for preventing or treating cancer, which includes N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition is excellent in treating cancer.

Description

PHARMACEUTICAL COMPOSITION FOR PREVENTING OR TREATING CANCER

[0001] The present invention relates to a pharmaceutical composition for preventing or treating cancer or a preparation containing a tyrosine kinase inhibitor compound and a use thereof.

[0002] According to the World Health Organization, about 20% of cancer patients die of lung cancer, accounting for the highest proportion of cancer-related mortality rates [CANCER TODAY 2022]. The cancers with the highest mortality rates following lung cancer are known to be gastric cancer (about 8% of cancer patients), liver cancer (about 8% of cancer patients), breast cancer (about 7% of cancer patients), and colorectal cancer (about 6% of cancer patients) [H Sung, CA 2021].

[0003] Meanwhile, lung cancer is histologically divided into two types (i.e., non-small cell lung cancer and small cell lung cancer) of which the former accounts for 85% of patients [Ferlay 2007]. Lung cancer is a highly heterogeneous carcinoma caused by a wide range of mutational abnormalities, including mutations in receptor tyrosine kinase (RTK). Targeted therapies have been approved for several RTK mutation-positive non-small cell lung cancers, but treatment options for some RTK mutations remain limited.

[0004] As the RTK mutations, a mutation in the human epidermal growth factor receptor 2 (HER2) tyrosine kinase domain, a mutation ofHER2amplification / overexpression, and an insertion mutation of an epidermal growth factor receptor (EGFR) exon 20 are known, and these mutations are potential therapeutic targets with distinct clinical characteristics.

[0005] The proportion of aHER2mutation as a cause of cancer is not high, accounting for around 5%. However, it has been reported that in most cancers where aHER2mutation is present, the mutation occurs in the tyrosine kinase domain, including exons 18 to 21 [Robichaux 2019]. A detailed analysis by cancer type of the protein domains in which aHER2mutation has occurred showed that the tyrosine kinase domain accounted for an average of 60% or more (65% for non-small cell lung cancer, 68% for breast cancer, and 68% for colorectal cancer). The mutation in theHER2tyrosine kinase domain that occurs in non-small cell lung cancer is an insertion mutation such as YVMA, VC, and GSP that occurs in exon 20; the mutation in theHER2tyrosine kinase domain that occurs in breast cancer is an L755S mutation that occurs in exon 19; and the mutation in theHER2tyrosine kinase domain that occurs in colorectal cancer is a V842I mutation that occurs in exon 21 [Robichaux 2019].

[0006] Additionally, in patients with non-small cell lung cancer, the mutation in theHER2tyrosine kinase domain accounts for 2-4%, andHER2amplification mutations account for 2-5%, respectively [Riudavets 2021]. Meanwhile, an EGFR Exon 20 insertion mutation account for 1.5-2.5% of all non-small cell lung cancers and appears in 6% of all non-small cell lung cancers with anEGFRmutation [Friedlaender 2022]. Representative subtypes ofEGFRexon 20 insertions observed in non-small cell lung cancer include SVD, ASV, NPH,etc. [Friedlaender 2022].

[0007] EGFRandHER2mutations have common characteristics, such as being frequently found in non-smokers, women, and patients with adenocarcinoma [Byeon 2019, Shigematsu 2005], and have been reported to have a poorer prognosis compared to other carcinogens [Mazieres 2013, Pillai 2017, Kris 2014, Yasuda 2012].

[0008] Additionally, about 30% of non-small cell lung cancer patients bearingHER2or anEGFRmutation are reported to develop intracranial disease and brain metastases during disease progression [Heon 2010]. Furthermore, brain metastases are found in 20% to 40% of cancer patients, and they are common in the natural progression of breast cancer, lung cancer, melanoma, renal cancer, and colorectal cancer, and occur particularly in 20% to 50% of patients with metastatic non-small cell lung cancer [Langer 2005]. The occurrence of brain metastases leads to a decline in quality of life, and the median survival in non-small cell lung cancer is only 4-6 months, and thus, highlighting the management of intracranial disease as a significant unmet need [Mehta 2003].

[0009] Meanwhile, there is an unmet need for new drugs that have strong therapeutic effects with fewer side effects in relation to the treatment of lung cancer, gastric cancer, colorectal cancer, breast cancer,etc., with aHER2orEGFRmutation.

[0010] For example, currently, the standard first-line treatment for NSCLC with aHER2mutations or an EGFR Exon 20 insertion mutation is a platinum-based chemotherapy, but trastuzumab-deruxtecan (product name Enhertu) is the only drug approved as a follow-up treatment after the standard first-line treatment for NSCLC with aHER2mutation. Additionally, amivantamab (product name Librivant) is the only drug approved as a follow-up treatment after the standard first-line treatment for non-small cell lung cancer with an EGFR Exon 20 insertion mutation.

[0011] However, these drugs have drawbacks in that they require intravenous administration and exhibit specific toxicity profiles causing side effects of infusion-related reactions and rash in the case of Librivant, and an increased risk of serious lung diseases including pneumonia and interstitial lung disease in the case of Enhertu [Kumagai 2020]. Therefore, there is still an unmet clinical need for novel drugs that exhibit a potent therapeutic effect with reduced side effects in relation to the treatment of lung cancer subtypes positive for aHER2mutation or an EGFR Exon 20 insertion mutation.

[0012] Moreover, unlike otherEGFRmutations in NSCLC an EGFR Exon 20 insertion mutation and aHER2mutations are generally known to confer resistance to the first-generation (gefitinib, erlotinib, icotinib) and the second-generation (afatinib, dacomitinib, neratinib)EGFRtyrosine kinase inhibitors due to steric hindrance within the drug-binding pocket. Due to this mechanism, it has been reported that drugs targeting cancers with an EGFR Exon 20 insertion mutation require higher plasma drug concentrations than are possible in clinical trials when considering dose-limiting toxicity to exhibit anti-cancer activity [Brazel 2022, Yasuda 2013]. Many studies have been conducted to develop novel tyrosine kinase inhibitors targeting an EGFR Exon 20 insertion mutation andHER2mutations, and recently developed exon 20-selective inhibitors, such as poziotinib and mobocertinib, have shown promising efficacy inHER2exon 20 insertions andEGFRexon 20 insertions, respectively. However, poziotinib is known to have safety issues, such as diarrhea, nausea, and rash, and showed limited efficacy against a HER2 Exon 20 insertion mutation [Kim 2018, Robichaux 2018)]. In addition, the FDA approval of mobocertinib was withdrawn because the primary endpoint (i.e., progression-free survival) was not met in a phase 3 clinical trial (EXCLAIM-2) after receiving accelerated approval from FDA.

[0013] As described above, aHER2mutation is also observed in patients with various solid cancers, including colorectal cancer, breast cancer, and gastric cancer, and the presence of aHER2mutation is known to change the tumor microenvironment [D Wang et a; / 2-22. Front. Immunol. 2022]. Additionally, mutations such asEGFRamplification are observed in colorectal cancer, breast cancer, and glioblastoma, and the presence of anEGFRmutation may decrease a response to drugs [ML Uribe et al. Cancers 2021]. Therefore, there is a need to develop a therapeutic agent that can exhibit a strong therapeutic effect on other solid tumors harboring aHER2mutation or anEGFRmutation that is expected to have a low therapeutic response to drugs.

[0014] One of the technical objects of the present invention provides a method for treating cancer in a subject having cancer and / or metastatic cancer, with an EGFR Exon 20 insertion mutation and / or a HER2 Exon 20 insertion mutation by includingEGFRaberration and / orHER2aberration, and particularly, a pharmaceutical composition for preventing or treating cancer or a preparation containing a pyrimidine derivative compound having anticancer activity as an active ingredient. Furthermore, the present invention provides a therapeutic method characterized by the dosage and / or administration method of the pyrimidine derivative to the subject.

[0015] Accordingly, the developers of the present invention, while making efforts to develop a cancer therapeutic agent that inhibits anEGFRmutation and aHER2mutation, have found that the pyrimidine derivative compound represented by Formula 1 of the present invention exhibits high inhibitory activity against anEGFRmutation and aHER2mutation when administered to a subject at a specific dosage, thus being useful for the prevention or treatment of cancer, and can also be used for the treatment of a subject with cancer harboring an EGFR Exon 20 insertion mutation and / or a HER2 Exon 20 insertion mutation that has metastasized to the brain, thereby completing the present invention.

[0016] [1] In one aspect of the present invention, the present invention relates to a pharmaceutical composition for preventing or treating cancer, which contains N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[0017] [Formula 1]

[0018]

[0019] [2] In Item [1] above, the daily dose of the active ingredient may be 5 mg to 240 mg.

[0020] [3] In Item [1] or [2] above, the pharmaceutical composition may be administered once a day, twice a day, or multiple times a day.

[0021] [4] In any one of Items [1] to [3] above, the pharmaceutical composition may be administered once a day.

[0022] [5] In any one of Items [1] to [3] above, the pharmaceutical composition may be administered twice a day.

[0023] [6] In Item [1] above, the active ingredient may be orally administered once or multiple times a day in a daily dose of 5 mg to 240 mg.

[0024] [7] In any one of Items [1] to [6] above, the pharmaceutical composition may be repeatedly administered every day.

[0025] [8] In any one of Items [1] to [7] above, the pharmaceutical composition may be orally administered.

[0026] [9] In any one of Items [1] to [8] above, the cancer may be a solid tumor.

[0027]

[0010] In any one of Items [1] to [9] above, the cancer may be selected from the group consisting of pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, bile duct cancer, large bowel cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus and nasal cavity cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestinal cancer, meningioma, esophageal cancer, glioma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoid, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational choriocarcinoma, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cancer, acoustic schwannoma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, ovarian cancer, and thymic cancer. The cancer of the subject may be one selected from the group consisting of lung cancer, breast cancer, colorectal cancer, gastric cancer, bladder cancer, pancreatic cancer, ovarian cancer, bile duct cancer, or maybe non-small cell lung cancer, gastric cancer, breast cancer, or colorectal cancer.

[0028]

[0011] In any one of Items [1] to

[0010] above, the cancer may haveEGFRaberration and / orHER2aberration.

[0029]

[0012] In Item

[0011] above, theEGFRaberration may beEGFRamplification and / orEGFRoverexpression, and theHER2aberration may beHER2amplification and / orHER2overexpression and / or a mutation of theHER2transmembrane or extracellular domain.

[0030]

[0013] In Item

[0012] above, the mutation of theHER2transmembrane or extracellular domain may be one selected from the group consisting of S310F, S310Y, R678Q, R678C, G309A, P122L, G222C, I655V, S305C, H470Q, I263T, and A293T.

[0031]

[0014] In any one of Items [1] to

[0010] above, the cancer may have anEGFRmutation and / or aHER2mutation.

[0032]

[0015] In Item

[0014] above, theEGFRmutation may be an EGFR Exon 20 insertion mutation, and / or theHER2mutation may be a mutation in theHER2tyrosine kinase domain and / or a mutation in theHER2transmembrane or extracellular domain.

[0033]

[0016] In Item

[0015] above, the mutation in theHER2tyrosine kinase domain may be an insertion mutation of exon 20 and / or a point mutation.

[0034]

[0017] In Item

[0015] above, the EGFR Exon 20 insertion mutation may include a mutation selected from the group consisting of del19, A763_Y764insFHEA, A763_Y764insFQEA, Y764_V765insHH, A767_S768insASV, S768dupSVD, V769_D770insASV, D770_N771insNPG, D770_N771insGL, D770_N771insSVD, D770_N771insSVG, DelD770insGY, DelD770insVG, N771_P772insH, N771_P772insV, DelN771insGY, DelN771insTH, P772_H773insDNP, P772_H773insPNP, H773_V774insNPH, H773_V774insPH, H773_V774insAH, H773_V774insH, and V774_C775insHV; and / or the mutation in theHER2tyrosine kinase domain may include a mutation selected from the group consisting of Y772dupYVMA, DelM774insWLV, A775_G776insYVMA, A775_G776insSVMA, A775_G776insI, DelG776insVC, DelG776insLC, G778_S779insCPG, V777_G778insGSP, G778dupGSP, P780_Y781insGSP, L755S, L755P, V777L, I767M, D769H, D769N, D769Y, G776V, G776S, V777M, and V842I; and / or the mutation in theHER2transmembrane or extracellular domain may include one selected from the group consisting of S310F, S310Y, R678Q, R678C, G309A, P122L, G222C, I655V, S305C, H470Q, I263T, and A293T.

[0035]

[0018] In Item

[0015] above, the cancer may have an EGFR Exon 20 insertion mutation.

[0036]

[0019] In Item

[0015] above, the cancer may have a mutation in theHER2tyrosine kinase domain and / or a mutation in theHER2transmembrane or extracellular domain.

[0037]

[0020] In any one of Items [1] to

[0019] above, the cancer may have metastasized to the brain of a subject.

[0038]

[0021] In any one of Items [1] to

[0020] above, the cancer may be a recurrent or refractoryHER2-positive cancer.

[0039]

[0022] In Item

[0021] above, the recurrent or refractoryHER2-positive cancer may be recurrent or refractoryHER2-positive breast cancer, recurrent or refractoryHER2-positive gastric cancer, or recurrent or refractoryHER2-positive colorectal cancer.

[0040]

[0023] In Item

[0021] or

[0022] above, the subject having recurrent or refractoryHER2-positive cancer may be a subject who has received treatment with aHER2inhibitor prior to the administration of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[0041]

[0024] In any one of Items

[0021] to

[0023] above, the subject having the recurrent or refractoryHER2-positive cancer may be a subject who has acquired resistance or refractory to treatment with aHER2inhibitor.

[0042]

[0025] In Item

[0023] or

[0024] above, theHER2inhibitor may be aHER2tyrosine kinase inhibitor.

[0043]

[0026] In Item

[0025] above, theHER2tyrosine kinase inhibitor may be one or more selected from the group consisting of afatinib, lapatinib, neratinib, and tucatinib.

[0044]

[0027] In Item

[0023] or

[0024] above, theHER2inhibitor may be an anti-HER2antibody or a drug conjugate thereof.

[0045]

[0028] In Item

[0027] above, the anti-HER2antibody or a drug conjugate thereof may be one or more selected from the group consisting of trastuzumab, trastuzumab emtansine, pertuzumab, margetuximab, and trastuzumab deruxtecan.

[0046]

[0029] In any one of Items

[0021] to

[0023] above, the subject having the recurrent or refractoryHER2-positive cancer may have received at least two prior treatments with differentHER2inhibitors prior to the administration of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[0047]

[0030] In any one of Items

[0021] to

[0029] above, the recurrent or refractoryHER2-positive cancer may be pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, bile duct cancer, large bowel cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus and nasal cavity cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestinal cancer, meningioma, esophageal cancer, glioma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoid, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational choriocarcinoma, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cancer, acoustic schwannoma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, ovarian cancer, and thymic cancer. The recurrent or refractoryHER2-positive cancer may be one selected from the group consisting of lung cancer, breast cancer, colorectal cancer, gastric cancer, bladder cancer, pancreatic cancer, ovarian cancer, and bile duct cancer.

[0048]

[0031] In another aspect of the present invention, the present invention provides a method for preventing or treating cancer, which includes administering, to a subject, an effective amount of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof:

[0049] [Formula 1]

[0050]

[0051]

[0032] In another aspect of the present invention, the present invention provides a use of an effective amount of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating cancer:

[0052] [Formula 1]

[0053]

[0054]

[0033] In another aspect of the present invention, the present invention provides a use of an effective amount of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof for preventing or treating cancer:

[0055] [Formula 1]

[0056]

[0057] The present invention provides a method for treating cancer by administering, to a subject, an effective amount of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, in particular preferably, N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition or a preparation therefor.

[0058] Fig. 1 is a drawing showing the tumor growth after 20 days of administration of control drug and Formula 1 in a Ba / F3HER2GSP subcutaneous implantation model.

[0059] Fig. 2 is a drawing showing the tumor growth after 24 days of oral administration of control drug and Formula 1 in a non-small cell lung cancer NCI-H1781HER2VC subcutaneous implantation model.

[0060] Fig. 3 is a drawing showing the tumor volume reduction ratios after 3 weeks of oral administration of Formula 1 in the non-small cell lung cancer NCI-H1781HER2VC intracerebral implantation model compared to the control drug administration.

[0061] Fig. 4 is a drawing showing the tumor growth after 20 days of administration of the control drug and Formula 1 in a Ba / F3HER2YVMA subcutaneous implantation model.

[0062] Fig. 5 is a drawing showing the tumor growth after 20 days of administration of the control drug and Formula 1 in a Ba / F3HER2VC subcutaneous implantation model.

[0063] Fig. 6 is a drawing showing the tumor growth after 28 days of administration of the control drug and Formula 1 in aHER2amplification / overexpression tumor model.

[0064] Fig. 7 is a drawing showing the tumor growth after 14 days of administration of the control drug and Formula 1 in a Ba / F3EGFRSVD subcutaneous implantation model.

[0065] Fig. 8 is a drawing showing the mean blood concentration-time profiles when Formula 1 was administered once intravenously (4 mg / kg) and when Formula 1 was administered once orally (5 mg / kg, 15 mg / kg, or 50 mg / kg) in mice.

[0066] Fig. 9 is a drawing showing the mean blood concentration-time profiles in rats when Formula 1 was administered once intravenously (IV, 2 mg / kg) of Formula 1 and when Formula 1 was administered once orally (PO, 2 mg / kg or 5 mg / kg).

[0067] Fig. 10 is a drawing showing the mean blood concentration-time profiles in beagle dogs when Formula 1 was administered once intravenously (IV, 1 mg / kg) and when Formula 1 was administered once orally (PO, 2 mg / kg).

[0068] Fig. 11 is a drawing showing the mean blood concentration-time profiles in mice when Formula 1 was orally administered once and repeatedly (PO) at 15 mg / kg or 50 mg / kg.

[0069] Fig. 12 is a drawing showing the results of pharmacokinetic modeling and simulation for predicting human clinical doses.

[0070] Definition

[0071] As used herein, the term“hydrate" refers to a compound of the present invention or a salt thereof containing a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.

[0072] As used herein, the term "solvate" refers to a compound of the present invention or a salt thereof containing a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces.

[0073] As used herein, the term "isomer" refers to a compound of the present invention or a salt thereof that has the same formula or molecular formula but is structurally or sterically different. The isomers include structural isomers (including tautomers) and stereoisomers. The stereoisomers include all of optical isomers (enantiomers) and diastereomers, and geometric isomers (trans,cis) that appear by having one or more asymmetric carbon centers. All isomers and mixtures thereof are included in the scope of the present invention.

[0074] As used herein, the term "aberration" refers to a change in the base sequence of a gene, an amplification or reduction in the number of genes, a deletion of a gene, or overexpression of a protein encoded by the corresponding gene.

[0075] As used herein, the term "pharmaceutically effective amount" or "effective dose" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment or improvement.

[0076] As used herein, the term "pharmaceutically acceptable" refers to a compound, material, composition, and / or dosage form that is suitable for use in contact with human tissue without causing excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment.

[0077] As used herein, the term "treatment" includes any effect that brings about improvement in a condition, disease, disorder,etc., such as alleviation, reduction, control, stabilization, improvement or elimination, or improvement of symptoms thereof. Specifically, for example, the treatment of cancer may refer to: (1) stabilization, reduction (e.g., greater than 10%, 20%, 30%, 40%, 50%, preferably greater than 60% of the cancer cell population and / or tumor size compared to those before administration) or elimination of cancer cells, (2) inhibition of cancer cell division and / or cancer cell proliferation, (3) alleviation to some extent (or preferably elimination) of one or more symptoms associated with the pathology associated with or caused in part by uncontrolled or abnormal cell division; (4) disease-free, relapse-free, progression-free, and / or an increase in the overall survival rate, duration, or ratio; (5) a decrease in hospitalization rate; (6) a decrease in the length of hospitalization; (7) eradication, removal, or control of primary, localized and / or metastatic cancer; (8) stabilization or reduction in the growth of tumor or neoplasm (e.g., by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, preferably by at least 80% compared to the initial growth rate); (9) impairment of tumor formation, (10) reduction in mortality, (11) an increase in the rate of response, durability of response, or number of patients showing a response or remission, (12) the tumor size is maintained and does not increase or increases by less than 10%, preferably less than 5%, preferably less than 4%, and preferably less than 2%; (13) reduction in the need for surgery (e.g., colectomy, mastectomy), and / or (14) prevention or reduction of metastasis of cancer cells.

[0078] As used herein, the term "administration" refers to any method that can be used to deliver an active ingredient to the site of desired biological action. Such methods of administration include, but are not limited to, an oral route, intraduodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, or infusion), topical / transdermal, and rectal / vaginal administration.

[0079] As used herein, the term "50% inhibition potency (IC50)", which is a measure of the potency of a substance to inhibit a specific biological or biochemical function, refers to a quantitative measurement representing the amount of a specific inhibitory substance (e.g., a drug) required to inhibit a given biological process or biological component by 50%in vitro.

[0080] As used herein, the term "Cmax" is the maximum (or peak) serum concentration of a drug in a specific compartment or test region of the body achieved after administration and prior to the administration of a second dose.

[0081] As used herein, the term "Tmax" is the time it takes for a drug that requires absorption to reach its maximum concentration (Cmax) after administration of the drug.

[0082] As used herein, the term "T1 / 2" is the time required for the plasma concentration of a drug to decrease by 50%.

[0083] As used herein, the term "Vdss" stands for volume of distribution in the body.

[0084] As used herein, the term "CL" is an indicator of the extent to which a drug is irreversibly eliminated from the circulation.

[0085] As used herein, the term "AUC0-last" is the area under the concentration-time curve from administration (time 0) to the last measured concentration.

[0086] As used herein, the term "AUC0-inf" is total drug exposure over time.

[0087] As used herein, the term "side effect" refers to any unfavorable or unintended disease or symptom that occurs in a patient receiving a drug.

[0088] A compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof

[0089] The compound represented by Formula 1 of the present invention is a tyrosine kinase inhibitor. The compound of Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, and a preparation method thereof are described in Korean Patent Application No. 10-2022-0093706, in which the contents of which are incorporated herein by reference in their entirety.

[0090] [Formula 1]

[0091]

[0092] The compound represented by Formula 1 above may be used directly (in free form) or in the form of a pharmaceutically acceptable salt.

[0093] The pharmaceutically acceptable salt of the compound represented by Formula 1 above is not particularly limited, and examples thereof include: addition salts with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid,etc.); addition salts with organic acids (e.g., acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane disulfonic acid, oxalic acid, isethionic acid,etc.); salts with alkali metals (e.g., potassium, sodium,etc.); salts with alkaline earth metals (e.g., calcium, magnesium,etc.); and a salt with an organic base such as ammonium salts, and ethylamine salts, alginates. The pharmaceutically acceptable salt may generally be synthesized by conventional chemical methods by reacting the compound represented by Formula 1 with a stoichiometric or sub-stoichiometric amount (e.g., 0.5 equivalents) of an appropriate base or acid in the presence of water or an organic solvent (e.g., ether, ethyl acetate, ethanol, isopropanol, or acetonitrile), or a mixture thereof.

[0094] The compound represented by Formula 1 above or a pharmaceutically acceptable salt thereof may be in the form of a solvate, which refers to a physical association of the compound represented by Formula 1 with one or more solvent molecules, regardless of whether it is organic or inorganic. Such a physical association includes a hydrogen bonding. In certain embodiments, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate may be isolated. The solvent molecules in a solvate may exist in a regular arrangement and / or non-ordered arrangement. The solvate may contain a stoichiometric or non-stoichiometric amount of solvent molecules. Solvates include both solution phases and isolable solvates. Exemplary solvent molecules capable of forming solvates include, but are not limited to, water, methanol, ethanol,n-propanol, isopropanol,n-butanol, isobutanol,tert-butanol, ethyl acetate, glycerin, acetone,etc.

[0095] A pharmaceutical composition or preparation containing a compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof

[0096] Another aspect of the present invention relates to a pharmaceutical composition or preparation for preventing or treating cancer or a preparation, which contains a compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[0097] In one embodiment of the present invention, the active ingredient contained in the pharmaceutical composition or preparation may be N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide or a pharmaceutically acceptable salt thereof.

[0098] In another embodiment of the present invention, the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be administered to a subject having cancer in a daily dose of 5 mg to 240 mg. In addition, the active ingredient may be administered, to a subject having cancer, at a dose of about 5 mg / day, about 10 mg / day, about 15 mg / day, about 20 mg / day, about 25 mg / day, about 30 mg / day, about 35 mg / day, about 40 mg / day, about 45 mg / day, about 50 mg / day, about 55 mg / day, about 60 mg / day, about 65 mg / day, about 70 mg / day, about 75 mg / day, about 80 mg / day, about 85 mg / day, about 90 mg / day, about 95 mg / day, about 100 mg / day, about 105 mg / day, about 110 mg / day, about 115 mg / day, about 120 mg / day, about 125 mg / day, about 130 mg / day, about 135 mg / day, about 140 mg / day, about 145 mg / day, about 150 mg / day, about 155 mg / day, about 160 mg / day, about 165 mg / day, about 170 mg / day, about 175 mg / day, 180 mg / day, about 185 mg / day, about 190 mg / day, about 195 mg / day, about 200 mg / day, about 205 mg / day, about 210 mg / day, about 215 mg / day, about 220 mg / day, about 225 mg / day, about 230 mg / day, about 235 mg / day, and up to about 240 mg / day.

[0099] Furthermore, the dose of the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, administered to the subject may be freely adjusted within a range such as about 5 mg / day to about 240 mg / day, about 20 mg / day to about 180 mg / day, and about 80 mg to about 120 mg / day. The dose adjustment may be freely changed during the administration period.

[0100] In an embodiment of the present invention, the pharmaceutical composition or preparation may be administered to the subject once a day or less, twice a day, or multiple times a day. Additionally, the daily dose of the pharmaceutical composition or preparation may be administered as a single dose or as multiple individually divided doses. For example, tablets or capsules, which contain 5 mg to 240 mg of the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, may be administered to a subject every other day (QOD), once daily (QD), or more than once daily (b.i.d., t.i.d.,etc.). The administration schedule may be determined based on sound medical judgment.

[0101] Meanwhile, the administration of the pharmaceutical composition or preparation may be performed for a specific treatment cycle, typically a treatment cycle of at least 28 days, regardless of the continuity of administration, according to the pharmacokinetics and drug elimination / accumulation of a specific patient. The administration may be repeated for several cycles with or without a drug-free period. The administration cycle may be longer or shorter, such as 7, 14, 18, 24, 28, 35, 42 days, or any range therebetween, and alternating or continuous day cycles may be used. Different administration schedules may be combined and used depending on the presence or absence of side effects, the cancer's response to treatment, patient convenience,etc.

[0102] In another embodiment of the present invention, the pharmaceutical composition or preparation may be administered repeatedly daily.

[0103] In still another embodiment of the present invention, the active ingredient contained in the pharmaceutical composition or preparation may be orally administered to a subject daily in a total daily dose of 5 mg to 240 mg.

[0104] In one embodiment of the present invention, the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be formulated into a dosage form suitable for oral administration. The dosage form suitable for oral administration includes, but is not limited to, a solid preparation or liquid-phase preparation.

[0105] Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, hard / soft capsules, boluses, granules, pastes for application to the tongue, preparations targeted for buccal or sublingual absorption, preparations targeted for systemic absorption, and troches. Liquid preparations for oral administration include, but are not limited to, suspensions, oral solutions, emulsions, emulsifiers, syrups, elixirs,etc.

[0106] Meanwhile, in formulating the compound represented by Formula 1 above, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, they may be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, surfactants,etc. The preparations above may be prepared by including at least one selected from the group consisting of diluents (e.g., water, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine), liquid paraffin, wetting agents, sweeteners, flavoring agents, preservatives, lubricants (e.g., silica, talc, stearic acid and a magnesium or calcium salt thereof, and / or polyethylene glycol), binders (e.g., magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidine), calcium carbonate, coloring agents, fragrances, and sweetening agents. Additionally, in some cases, it may contain a disintegrating agent or boiling mixture and / or an absorbent, such as starch, agar, alginic acid or a sodium salt thereof.

[0107] In one embodiment of the present invention, as the drug product of the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be an immediate-release capsule preparation, and the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be contained as a drug substance. The drug product of the compound represented by the above Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be prepared in strengths of 5 mg, 30 mg, or 60 mg, and these drug products may be formulated as capsules containing white opaque hydroxypropyl methylcellulose (HPMC) containing white to off-white powder.

[0108] The compound represented by the Formula 1 of the present invention, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or preparation containing the same may be administered to the subject in a pharmaceutically effective amount or effective dose. The pharmaceutically effective amount may be determined based on factors including the type and severity of the disease, age, sex, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently-used drugs, and other factors well known in the medical field. For example, the pharmaceutically effective amount or effective dose of the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be from 0.01 mg / kg to 75 mg / kg.

[0109] In another embodiment of the present invention, a pharmaceutical composition or preparation for preventing or treating cancer containing the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient may enhance the anticancer effect by administering individually as a therapeutic agent or by administering in combination with another anticancer agent in use.

[0110] Meanwhile, the anticancer agent that can be combined with the compound represented by the above Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be an activator that interferes with or inhibits the RAS-RAF-ERK or PI3K-AKT-TOR signaling pathway, or is a PD-1 and / or PD-L1 antagonist. Additionally, immunotherapies including RAF inhibitors,EGFRinhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors, or monoclonal antibodies, immunomodulatory imides (IMiDs), anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAGl, and anti-OX40 agents, GITR agonists, CAR-T cells, and BiTEs may be combined with the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, but are not limited thereto.

[0111] RAF inhibitors include dabrafenib, encorafenib, regorafenib, sorafenib, and vemurafenib, but are not limited thereto.

[0112] MEK inhibitors include binimetinib, CI-1040, cobimetinib, PD318088, PD325901, PD334581, PD98059, refametinib, selumetinib, and trametinib, but are not limited thereto.

[0113] ERK inhibitors include LY3214996, LTT462, MK-8353, SCH772984, raboxertinib, ulixertinib, and ASTX029, but are not limited thereto.

[0114] PI3K inhibitors include 17-hydroxybortmannine analogs (e.g., WO 06 / 044453); AEZS-136; alpelisib; AS-252424; buparlisib; CAL263; copanlisib; CUDC-907; dactolisib (e.g., WO 06 / 122806); demethoxyviridin; duvelisib; GNE-477; GSK1059615; IC87114; idelalisib; INK1117; LY294002; palomid 529; paxalisib; perifosine; PI-103; PI-103 hydrochloride; pictilisib (e.g., WO 09 / 036,082; WO 09 / 055,730); PIK 90; PWT33597; SF1126; sonolisib; TGI 00-115; TGX-221; XL147; XL-765; wortmannin; taselisib (GDC-0032); and ZSTK474, but are not limited thereto.

[0115] AKT inhibitors include Akt-1-1 (inhibiting Aktl) (Barnettet al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (Barnettet al. (2005) Biochem. J. 385 (Pt. 2), 399-408); API-59CJ-Ome (e.g., Jinet al. (2004) Br. J. Cancer 91, 1808-12); l-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05011700); Indole-3-carbinol and derivatives thereof (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. 134(12 Suppl), 3493S-3498S); perifosine (Dasmahapatraet al. (2004) Clin. Cancer Res. 10(15), 5242-52, 2004); phosphatidylinositol ether lipid analogues (e.g., Gills and Dennis (2004) Expert. Opin. Investig. Drugs 13, 787-97); triciribine (Yanget al. (2004) Cancer Res. 64, 4394-9); Imidazooxazone compounds including trans-3-amino-1-methyl-3-[4-(3-phenyl-5H-imidazo[1,2-c]pyrido[3,4-e][1,3]oxazin-2-yl)phenyl]-cyclobutanol hydrochloride (WO 2012 / 137870); apuresertib; capivasertib; 8-[4-(1-aminocyclobutyl)phenyl]-9-phenyl-1,2,4-triazolo[3,4-f][1,6]naphthyridin-3(2H)-on (MK2206) and pharmaceutically acceptable salts thereof; AZD5363; trans-3-amino-1-methyl-3-(4-(3-phenyl-5H-imidazo[1,2-c]pyrido[3,4-e][1,3]oxazin-2-yl)phenyl)cyclobutanol (TAS117) and pharmaceutically acceptable salts thereof; and patacertib, but are not limited thereto.

[0116] TOR inhibitors include deforolimus; ATP competitive TORC1 / TORC2 inhibitors including PI-103, PP242, PP30, and Torin 1; TOR inhibitors of the FKBP12 enhancer, rapamycin, temsirolimus, everolimus, those disclosed in WO 94 / 09010 and derivatives thereof; rapalogs, for example those disclosed in WO 98 / 02441 and WO 01 / 14387, for example AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin; 40-epi-(tetrazolyl)-rapamycin (or ABT578); AZD8055; 32-deoxorapamycin; 16-pentynyloxy-32(S)-dehydrorapannacin and other derivatives disclosed in WO 05 / 005434; derivatives disclosed in US 5,258,389, WO 94 / 090101, WO 92 / 05179, US 5,118,677, US 5,118,678, US 5,100,883, US 5,151,413, US 5,120,842, WO 93 / 111130, WO 94 / 02136, WO 94 / 02485, WO 95 / 14023, WO 94 / 02136, WO 95 / 16691, WO 96 / 41807, WO 96 / 41807, and US 5,256,790; and rapamycin derivatives containing phosphorus (e.g., WO 05 / 016252), but are not limited thereto.

[0117] MCL-1 inhibitors include AMG-176, MIK665, and S63845, but are not limited thereto.

[0118] SHP2 inhibitors include the SHP2 inhibitors disclosed in JAB-3068, RMC-4630, TNO155, SHP-099, RMC-4550, WO 2019 / 167000, WO 2020 / 022323, and WO 2021 / 033153, but are not limited thereto.

[0119] RAS inhibitors include AMG510, MRTX849, LY3499446, JNJ-74699157(ARS-3248), ARS-1620, ARS-853, RM-007, and RM-008, but are not limited thereto.

[0120] In addition, as an anticancer agent that can be combined with the compound represented by the above Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof 2-ethylhydrazide, 2,2',2"-trichlorotriethylamine, ABVD, aceglatone, acemannan, aldophosphamide glycoside, alpharadin, amifostine, aminolevulinic acid, anagrelide, ANCER, ancestim, anti-CD22 immunotoxin, antitumorigenic herbs, apaziquone, arglabine, arsenic trioxide, azathioprine, BAM 002 (Novelos), bcl-2 (Genta), bestrabucil, viricor, bisanthrene, bromocriptine, brostalysin, bryostatin, buthionine sulfoximine, calyculin, cell-cycle nonspecific antineoplastic agents, celmoleukin, clodronate, clotrimazole, Cytarabine ocfosfate, DA 3030 (Dong-A), defopamine, denileukin diftitox, dexrazoxane, diaziquone, dichloroacetic acid, dilazep, discodermolide, docosanol, doxercalciferol, edelfosine, eflornithine, EL532 (Elan), elfomitine, elsamitrucine, eniluracil, etanidazole, exiserind, peruzinol, folic acid supplements (e.g., proline), gacytosine, gallium nitrate, a gimeracil / oteracil / tegafur combination (S-1), glycopine, histamine dihydrochloride, HIT diclofenac, HLA-B7 gene therapy (Vical), human fetal alpha-fetoprotein, ibandronate, ibandronic acid, ICE chemotherapy regimen, imexone, iobenguan, IT-101 (CRLX101), raniquidar, LC 9018 (Yakult), leflunomide, lentinan, levamisole + fluorouracil, lovastatin, lucanthone, masoprocol, melarsoprol, metoclopramide, miltefosine, miproxifen, mitoguazone, mitozolomide, mopidamol, motexapine gadolinium, MX6 (Galderma), naloxone + pentazocine, nitracrine, nolatrexed, NSC 631570 octreotide (Ukraine), olaparib, P-30 protein, PAC-1, palifermin, pamidronate, pamidronic acid, sodium pentosan polysulfate, phenamet, picivanil, pixantrone, platinum, podophyllic acid, porfimer sodium, polysaccharide-K (PSK), rabbit antithymocyte polyclonal antibody, rasburiemboriment, retinoic acid, rhenium Re 186 etidronate, romurtide, samarium (153 Sm) lexidronam, sizopyran, sodium phenylacetate, sparfosic acid, spirogermanium, strontium-89 chloride, suramin, swainsonine, talaporfin, tariquidar, tazarotene, tegafur-uracil, temoporfin, tenuaginic acid, tetrachlorodecaoxide, thrombopoietin, tin ethyl etiofurpurine, tirapazamine, TLC ELL-12, tositumomab-iodine 131, a combination of trifluridine and tipiracil, troponin I (Harvard University, USA), urethane, valspodar, verteporfin, zoledronic acid, and zosuquidar may be used.

[0121] As used herein, the term "combination" refers to a therapy involving the use of two or more compounds / drugs in combination. "Combination" may refer to compounds or drugs that are administered together for the entire administration period or for a certain period of time. When two or more compounds / drugs are administered in combination, the doses of each may be different. Combinations include administering the compounds / drugs in a sequential manner (i.e., wherein each compound / drug is administered at different times) as well as administering the compounds / drugs, or at least two of the compounds / drugs in a substantially simultaneous manner. For example, the substantial simultaneous administration may be achieved by administering to a subject a single dosage form having a fixed ratio of each compound / drug, or multiple single dosage forms for each compound / drug. The sequential or substantially simultaneous administration of each compound / drug may be performed by any suitable route, including but not limited to an oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissues (e.g., buccal). The compounds / drugs may be administered by the same route or by different routes. For example, the first compound / drug of the selected combination may be administered by intravenous injection, while the other compounds / drugs of the combination may be administered orally. Alternatively, for example, all compounds / drugs may be administered orally or all compounds / drugs may be administered by intravenous injection.

[0122] The combinations may include those used in conjunction with other biologically active ingredients and non-pharmacological therapies (e.g., surgery or radiotherapy). When the combination therapy further includes a non-pharmacological treatment, the non-pharmacological treatment may be administered at any appropriate time as long as a beneficial effect is achieved from the synergistic action of the combination of the compound / drug and the non-pharmacological treatment. The radiation therapy may be performed by one or a combination of several methods including but not limited to external-beam therapy, internal radiation therapy, implant radiation, stereotactic radiosurgery, total body radiation therapy, radiotherapy, and permanent or temporary brachytherapy. The "brachytherapy" means radiation therapy delivered by a spatially confined radioactive material inserted into the body at or near the site of a tumor or other proliferative tissue disease. The brachytherapy may include, but is not limited to, exposure to radioisotopes (e.g., radioisotopes of At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and Lu).

[0123] In one embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or preparation is administered may be a solid tumor. The cancer or solid tumor may be selected from the group consisting of pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, bile duct cancer, large bowel cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus and nasal cavity cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestinal cancer, meningioma, esophageal cancer, glioma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoid, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational choriocarcinoma, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cancer, acoustic schwannoma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, ovarian cancer, and thymic cancer, but is not limited thereto.

[0124] In another embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or preparation is administered may be one selected from the group consisting of lung cancer, breast cancer, colorectal cancer, gastric cancer, bladder cancer, pancreatic cancer, ovarian cancer, and bile duct cancer.

[0125] In still another embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or preparation is administered may be non-small cell lung cancer, gastric cancer, breast cancer, or colorectal cancer.

[0126] In one embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or preparation is administered may be a cancer harboringEGFRaberration and / orHER2aberration or anEGFR-positive and / orHER2-positive cancer. In this case, "EGFR-positive" means a cancer in whichEGFRprotein and / orEGFRgene is detected / detectable, and similarly, "HER2-positive" means a cancer in whichHER2protein and / orHER2gene is detected / detectable.

[0127] The "aberration" ofEGFRand / orHER2refers to a gain-of-function change that causes or contributes to cancer formation and / or development, such as protein overexpression, gene amplification (e.g., a change in copy number), activating gene mutation / activating protein mutation (e.g., insertion, point, or deletion mutation), activating chromosomal translocation / insertion / inversion, activating gene rearrangement or gene fusion (a subset of gene rearrangements), misregulation / dysregulation,etc. (including combinations thereof). For example, a cancer harboring anEGFRand / orHER2aberration can express (i) amplified or overexpressedEGFRand / orHER2, including that of a wild-type variant; (ii) constitutively activatedEGFRand / orHER2(including that of a wild-type variant); (iii)HER2with one or more mutations in the transmembrane or extracellular domain regions; or (iv) a combination thereof.

[0128] Additionally, there are currently no generally applicable validated diagnostic criteria forEGFRoverexpression or amplification. However, it refers to a case in which they were confirmed by one or more of immunohistochemistry (IHC),in situhybridization (ISH), and nucleic acid-based diagnostic tests, and diagnosed by applying the cut-off criteria of the corresponding testing institution in a clinical laboratory that is Clinical Laboratory Improvement Amendment (CLIA) accredited (in the United States) or equivalents thereof (areas outside the United States).

[0129] Additionally, theHER2overexpression refers to a case where the test result of immunohistochemistry (IHC) is 3+, andHER2amplification refers to a case where the test result of dual-probe fluorescencein situhybridization isHER2 / CEP17 ratio ≥ 2.0 orHER2 / CEP17 ratio < 2.0 with an averageHER2copy number ≥ 6.0 signals / cell.

[0130] Additionally, theHER2aberration may be one or more mutations in the transmembrane or extracellular domain region ofHER2, examples of which include but are not limited to S310X (where X is any amino acid) exemplified by S310F and S310Y; R678X (where X is any amino acid) exemplified by R678Q and R678C, as well as G309A, P122L, G222C, I655V, S305C, H470Q, I263T, and A293T.

[0131] Mutations in theHER2protein may be those described in US patent application US 2021 / 0024530, the contents of which are incorporated herein by reference in their entirety.

[0132] In a specific embodiment of the present invention, theEGFRaberration may beEGFRamplification and / orEGFRoverexpression, and theHER2aberration may beHER2amplification and / orHER2overexpression and / or a mutation in theHER2transmembrane or extracellular domain. The aberration ofEGFRand / orHER2may be a form of constitutively activatedEGFRand / orHER2(including wild-type mutants) due to abnormal ligand binding.

[0133] Unless otherwise stated, all references regardingEGFRamino acid sequence information are based on human wild-typeEGFRisoform a, which is accessible from the National Center for Biotechnology Information (NCBI) protein database under accession numbers NP-005219.2, P00533.2,etc. Unless otherwise stated, all references regardingHER2amino acid sequence information are based on human wild-typeHER2isoform a, which is available from the National Center for Biotechnology Information (NCBI) protein database under accession numbers NP_004439.2, NM_004448,etc.

[0134] Isoforms ofEGFRandHER2are also known to those skilled in the art, and the present disclosure also encompasses these isoforms. The term "isoform" means that transcripts produced at the same locus of a gene can potentially induce different functions in different forms of mRNAs, transcription start sites (TSSs), protein coding DNA sequences (CDSs), and untranslated regions (UTRs).

[0135] In another embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or preparation is administered may have anEGFRmutation and / or aHER2mutation. Meanwhile, the pharmaceutical composition or preparation may be able to inhibit theEGFRmutation and / or theHER2mutation and may have the activity of growth inhibition, prevention, improvement, or treatment of cancer.

[0136] In still another embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or preparation is administered may have anEGFRmutation or a mutation in theEGFRtyrosine kinase domain.

[0137] TheEGFRmutation may be located in the tyrosine kinase domain ofEGFR, and may include, but is not limited to, one or more of an exon 18 mutation (in the region of 688-728); an exon 19 mutation (in the region of 729-761); an exon 20 mutation (in the region of 762-823); or an exon 21 mutation (in the region of 824-875).

[0138] TheEGFRexon 18 mutations may include, but are not limited to, point mutations, such as E709X or G719X (where X is any amino acid) exemplified by E709K, E709A, E709G, G719A, G719S, and G719C, deletion mutations, and deletion-insertion mutations. The mutations may include, but are not limited to, deletions of glutamic acid at position 709 and threonine at position 710, insertion of aspartic acid (DelE709_T710insD),etc.

[0139] TheEGFRexon 19 mutations may include not only "typical" exon 19 deletion mutations of at least three amino acid residues, but also deletion-insertion mutations,etc. The mutations may include, but are not limited to, DelE746_A750 (deletion from glutamic acid at position 746 to alanine at position 750), DelL747_P753insS (deletion from leucine at position 747 to proline at position 753 and insertion of serine), DelE746_T751insA, DelE746_S752insD, DelL747_T751, DelL747_A750insP,etc.

[0140] TheEGFRexon 20 mutations may include, but are not limited to, point mutations (e.g., T790M, S768I, V769M, and H773R), deletion mutations, and insertion mutations (SVD, ASV, and NPH). In particular, the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof has activity against cancers harboring one or more insertion mutations ofEGFRexon 20. An EGFR Exon 20 insertion mutation is found at a relatively high prevalence not only in non-small cell lung cancer (NSCLC) but also in squamous cell carcinoma of the nasal cavity (NSCC), and is associated with novel resistance to currently clinically availableEGFRinhibitors, thus making it a desirable target for the treatment in the present application.

[0141] The EGFR Exon 20 insertion mutation may be a heterologous in-frame insertion of 1 to 7 amino acids (indicated as "insX") spanning about 15 amino acids (D761 to C775) of exon 20. EGFR Exon 20 insertion mutation may be D761_E762insX (insertion of 1 to 7 amino acid residues "X" between aspartic acid at position 761 and glutamic acid at position 762), A763_Y764insX, Y764_V765insX, V765_M766insX, A767_S768insX, S768_V769insX, V769_D770insX, D770_N771insX, N771_P772insX, P772_H773insX, H773_V774insX, or V774_C775insX. Additionally, the mutations may include deletion-insertion mutations such as DelD770insX (deletion of aspartic acid at position 770 and insertion of 1 to 7 amino acids "X") and DelN771insX (Simon Vyse and Paul H. Huang, TargetingEGFRexon 20 insertion mutations in non-small cell lung cancer. Signal Transduct Target Ther. 2019 Mar 8;4:5).

[0142] The EGFR Exon 20 insertion mutation may include del19, A763_Y764insFHEA, A763_Y764insFQEA, Y764_V765insHH, A767_S768insASV, S768dupSVD, V769_D770insASV, D770_N771insNPG, D770_N771insGL, D770_N771insSVD, D770_N771insSVG, DelD770insGY, DelD770insVG, N771_P772insH, N771_P772insV, DelN771insGY, DelN771insTH, P772_H773insDNP, P772_H773insPNP, H773_V774insNPH, H773_V774insPH, H773_V774insAH, H773_V774insH, and V774_C775insHV, but are not limited thereto (Takayuki Kosakaet al. Response Heterogeneity ofEGFRandHER2Exon 20 Insertions to CovalentEGFRandHER2Inhibitors. Cancer Res. 2017 May;77(10):2712-2721).

[0143] In one embodiment of the present invention, the aberration ofHER2may be in the form of gene amplification and / or protein overexpression. The aberration ofHER2may be in the form of one or more mutations of theHER2protein.

[0144] In one embodiment of the present invention, theHER2mutation may be a mutation in theHER2tyrosine kinase domain and / or a mutation in theHER2transmembrane or extracellular domain. The types of the mutation in theHER2transmembrane or extracellular domain are as described above.

[0145] In another embodiment of the present invention, the mutation in theHER2tyrosine kinase domain may be located in the tyrosine kinase domain ofHER2, and may include, but not limited to, one or more mutations selected from exon 19 mutation (in the region of 736-769); exon 20 mutation (in the region of 770-831) or exon 21 mutation (in the region of 832-883); and exon 22-31 (in the region of 884-1255).

[0146] TheHER2exon 19 mutations may include point mutations, deletion mutations (e.g., DelL755_T759),etc., but not limited thereto. TheHER2exon 19 point mutations may include L755X, I767X, and D769X (where X is any amino acid), exemplified by L755S, L755P, I767M, D769H, D769N, and D769Y.

[0147] TheHER2exon 20 mutations may include, but are not limited to, point mutations, deletion mutations, and insertion mutations. TheHER2exon 20 point mutations may include G776X and V777X (where X is any amino acid), exemplified by G776V, G776S, V777L, and V777M.

[0148] Meanwhile, the compound represented by Formula 1 in the present Example, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof was surprisingly found to exhibit anticancer effects in cancers harboring one or more insertion mutations ofHER2exon 20. The HER2 Exon 20 insertion mutation is found in various cancer types, particularly in lung cancer (e.g., NSCLC) and breast cancer, but treatment of the HER2 Exon 20 insertion mutation remains a clinical challenge and afatinib, lapatinib, and neratinib, which are approvedHER2TKIs, have limited efficacy in this patient group. Therefore, the HER2 Exon 20 insertion mutation is a desirable target for the treatment in the present application.

[0149] The HER2 Exon 20 insertion mutation may be a heterologous in-frame insertion of 1 to 7 amino acids in A775_G776insX of exon 20 (insertion of 1 to 7 amino acid residues "X" between alanine at position 775 and glycine at position 776), a duplication insertion mutation such as Y772dupYVMA (duplication of the YVMA sequence starting at tyrosine at position 772), and a deletion-insertion mutation such as DelG776insX (deletion of glycine and insertion of 1 to 7 amino acids "X" at position 776). The HER2 Exon 20 insertion mutation includes Y772dupYVMA, DelM774insWLV, A775_G776insYVMA, A775_G776insSVMA, A775_G776insI, DelG776insVC, DelG776insLC, G778_S779insCPG, V777_G778insGSP, G778dupGSP, P780_Y781insGSP, L755S, L755P, V777L, or V842I, but is not limited thereto (Takayuki Kosakaet al. Response Heterogeneity ofEGFRandHER2Exon 20 Insertions to CovalentEGFRandHER2Inhibitors. Cancer Res. 2017 May;77(10):2712-2721; Jacqulyne P Robichauxet al. Pan-Cancer Landscape and Analysis of ERBB2 Mutations Identifies Poziotinib as a Clinically Active Inhibitor and Enhancer of T-DM1 Activity. Cancer Cell. 2019 Oct;36(4):444-457).

[0150] In one embodiment of the present invention, theEGFRmutation may be an EGFR Exon 20 insertion mutation, and theHER2mutation may be a mutation in theHER2tyrosine kinase domain and / or a mutation in theHER2transmembrane or extracellular domain. In addition, the mutation in theHER2tyrosine kinase domain may be an insertion mutation of exon 20 and / or a point mutation.

[0151] In another embodiment of the present invention, embodiments of the EGFR Exon 20 insertion mutation, the mutation in theHER2tyrosine kinase domain, and the mutation in theHER2transmembrane or extracellular domain are as described above.

[0152] In another embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or formulation is administered may have an EGFR Exon 20 insertion mutation. Additionally, the cancer may have a mutation in theHER2tyrosine kinase domain and / or a mutation in theHER2transmembrane or extracellular domain.

[0153] In one embodiment of the present invention, the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof has high brain penetrability. Accordingly, cancers that can be treated with the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof include metastatic brain tumors, for example brain metastases of any of the cancer types above (e.g., lung cancer, breast cancer, gastric cancer, bladder cancer, and bile duct cancer including gallbladder cancer and cholangiocarcinoma), which may have metastasized to the brain.

[0154] In one embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or preparation is administered may be a brain metastasis. For example, the cancer may be a metastatic brain tumor that has metastasized from one of the above-mentioned solid tumors as a primary cancer.

[0155] Preferred embodiments of the present disclosure relate to treating a subject having cancer, which includes treating a solid tumor havingEGFRaberration and / orHER2aberration (e.g., lung cancer, breast cancer, colorectal cancer, gastric cancer,etc.), particularly when the cancer has metastasized to the brain.

[0156] In another embodiment of the present invention, the cancer of the subject to which the pharmaceutical composition or formulation is administered, may be recurrent or refractoryHER2-positive.

[0157] In another embodiment of the present invention, the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof may be particularly useful in the treatment of unresectable advanced (stage III) and metastatic (stage IV) disease, "recurrent" and "refractory" cancers, or cancers that have not responded to medical treatment to date. A "recurrent" cancer is a cancer that relapses (comes back) after a certain period of time when it was usually undetectable. The cancer may return in the same location as the original (primary) tumor or in a different location in the body. The "refractory" cancers may appear as resistant / refractory from the start, or resistance / refractory may be acquired by the cancer cells during the course of prior therapy, and thus may include recurrent cancers that initially respond to treatment but often relapse in a more aggressive / resistant form. For example, the cancer may be a recurrent or refractoryHER2-positive cancer, preferably a recurrent or refractory cancer in whichHER2is genetically amplified and / or overexpressed. Exemplary cancer types may include recurrent or refractoryHER2-positive breast cancer, recurrent or refractoryHER2-positive gastric cancer, or recurrent or refractoryHER2-positive colorectal cancer, but are not limited thereto.

[0158] The subject having the recurrent or refractoryHER2-positive cancer may be a subject who has received treatment with aHER2inhibitor prior to administering N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof. In particular, the subject who has previously received treatment using aHER2inhibitor may be a subject who has acquired resistance or refractory to treatment using theHER2inhibitor. For example, the subject withHER2-positive cancer who has been previously treated with one or more anticancer agents but has not responded to or has relapsed following prior treatment(s) with the anticancer agent(s) may exhibit resistance / refractory properties as a result of exposure of the cancer to the anticancer agent(s). The resistance / refractory property may be observed in cancers withEGFRand / orHER2aberration. Cancers with theEGFRand / orHER2aberrations may appear in the form ofEGFRand / orHER2overexpression and / or mutations or any other cancer-causing alterations resulting in loss of function of tumor suppressor genes / proteins or gain-of-function change of oncogenes / oncogene-encoding proteins.

[0159] In another embodiment of the present invention, theHER2inhibitor may be aHER2tyrosine kinase inhibitor, an anti-HER2antibody, or a drug conjugate thereof. In particular, theHER2inhibitor may be one or more selected from the group consisting of afatinib, lapatinib, neratinib, and tucatinib, but is not limited thereto. Additionally, the anti-HER2antibody or drug conjugate thereof may be one or more selected from the group consisting of trastuzumab, trastuzumab emtansine, pertuzumab, margetuximab, and trastuzumab deruxtecan, but is not limited thereto.

[0160] In another embodiment of the present invention, the subject having the recurrent or refractoryHER2-positive cancer may be a subject who has received at least two treatments with differentHER2inhibitors prior to administering the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof; and the cancer of the subject may be one of the solid tumors described above, or one selected from the group consisting of lung cancer, breast cancer, colorectal cancer, gastric cancer, bladder cancer, pancreatic cancer, ovarian cancer, and bile duct cancer.

[0161] Additionally, prior to starting treatment with the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, it may be determined whether the subject has one or more aberrations inEGFRand / orHER2as confirmed above. For example, it may involve a pre-screening step to determine whether the subject has aberrations inEGFRand / orHER2and if they are good candidates for treatment. The aberrations may be determined by analyzing the subject's genotype or by analyzing any biological sample, including a blood or tumor sample, taken from the subject using the methods described below, from a family history of cancer involving the aberrations, or from past records or previous tests performed on the subject. When the subject is determined to beEGFR-positive and / orHER2-positive and has one or more aberrations as described in the present disclosure, it is possible to apply a treatment using the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[0162] The presence ofEGFRand / orHER2aberrations (including, for example, aberrations that induce protein activation through abnormal ligand binding) may be determined, for example, during prescreening of a subject or through previous tests performed on the subject, or maybe confirmed according to known assays, including licensed or approvedin vitrodiagnostic (IVD) assays or assays designed for this purpose. Examples include, but are not limited to, next generation sequencing (NGS)-based gene panels, whole exome profiling, polymerase chain reaction (PCR),in situhybridization (ISH), immunohistochemistry (IHC), flow cytometry, or other assays that can determineEGFRand / orHER2aberrations in tumor tissue or circulating tumor DNA (ctDNA), RNA, protein,etc. For example, a subject without a recorded tumor tissue sample may be biopsied, and a fresh tumor biopsy may be analyzed to identify any underlying aberrations.

[0163] Use of the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof

[0164] An aspect of the present invention relates to a method for preventing or treating cancer, which includes administering to a subject an effective amount of the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, and the above-described contents with respect to the pharmaceutical composition or preparation containing the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof apply equally.

[0165] An aspect of the present invention relates to the use of an effective amount of the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof in preparation of a medicament for preventing or treating cancer, and the above-described contents with respect to the pharmaceutical composition or preparation containing the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof apply equally.

[0166] Another aspect of the present invention relates to the use of an effective amount of the compound represented by the Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof for preventing or treating cancer, and the above-described contents with respect to the pharmaceutical composition or preparation containing the compound represented by Formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof apply equally.

[0167] Hereinafter, the present invention will be described in more detail through examples. These examples are only intended to explain the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention.

[0168] Preparation Example 1. Preparation of drug product including N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide (hereinafter referred to as Formula 1

[0169] The drug product used in one embodiment of the present invention is a fast-release capsule preparation containing a raw drug substance, Formula 1. The preparation contains Formula 1 in strengths of 5 mg, 30 mg, or 60 mg. In particular, the 5 mg content preparation is a white opaque hydroxypropyl methylcellulose (HPMC) No. 0 capsule containing a white to off-white powder; the 30 mg content preparation is a light blue opaque HPMC No. 0 capsule containing white to off-white powder; and the 60 mg content preparation is an orange opaque HPMC No. 0 capsule containing white to off-white powder.

[0170] Example 1. Evaluation ofin vitroactivity of Formula 1 againstHER2orEGFRmutation

[0171] In order to confirm the inhibitory potency and mutation coverage of Formula 1 onHER2andEGFRmutations, Ba / F3 cells engineered to express aHER2orEGFRmutation were treated with Formula 1 at concentrations ranging from 100 pM to 1 μM (prepared by approximately 3.16-fold serial dilutions) or a control drug for 72 hours. Then, ATP was quantified using CellTiter Glo (Promega), and the 50% inhibition of cell proliferation (IC50) was calculated, and the results are shown in Table 1.

[0172]

[0173]

[0174] As shown in Table 1, it was confirmed that Formula 1 showed strong inhibitory activity against 13 major mutation subtypes occurring in theHER2tyrosine kinase domain region (an HER2 Exon 20 insertion mutation and a point mutation ofHER2tyrosine kinase), and among these, inhibitory activities of about 1.5 to 4.5-fold higher than zongertinib, 19 to 72-fold higher than tucatinib, and 18 to 94-fold higher than lapatinib were exhibited for the YVMA, SYMA, VC, and GSP subtypes, which are insertion mutations ofHER2exon 20, whereas for L755S, L755P, D769N, V773M, V777L, T798M, L869R, L869R / T798I, and V842I subtypes, inhibitory activities in the range of about 7.7-fold to 500-fold or higher than tucatinib or lapatinib, thus being superior to the control drug.

[0175] In addition, it was confirmed that Formula 1 is effective in SVD, ASV, NPH, GY, HH,etc., which are insertion mutations ofEGFRexon 20, and in particular, the activity was calculated to be at least 10.3-fold higher than osimertinib and 1.5 to 3.5-fold higher than sunvozertinib for major subtypes such as SVD, ASV, and NPH, thus exhibiting a superior level of effect. In addition, it was confirmed that Formula 1 has strong activity with an IC50of about 1 nM in theHER2wild type.

[0176] In summary, it was shown that Formula 1 exhibited excellent inhibitory activity against all major subtypes of mutations in theHER2tyrosine kinase domain, indicating a broad spectrum of activity. In addition, Formula 1 exhibited an inhibitory effect similar to or higher than that of the control drug against the major subtypes of an EGFR Exon 20 insertion mutation. Therefore, Formula 1 may be effective in treating patients with solid tumors harboring mutations in theHER2tyrosine kinase domain and an EGFR Exon 20 insertion mutation.

[0177] Example 2. Evaluation ofin vitroactivity of Formula 1 againstHER2amplification / overexpression

[0178] In order to confirm the anticancer effect of Formula 1 in wild-typeHER2amplified / overexpressing NCI-N87 gastric cancer cells, Formula 1 was treated with the NCI-N87 cell line at concentrations ranging from 51 pM to 1 μM (prepared by 3-fold serial dilutions) for 72 hours. Comparator compounds (zongertinib, mobocertinib, lapatinib, or tucatinib) with known activity againstHER2-positive cancer cells were tested under the same conditions. Cell viability was measured using ATPLite (PerkinElmer). The results are shown in Table 2 below.

[0179] Genetic MutationIC50(nM)Formula 1ZongertinibMobocertinibLapatinibTucatinibAmplification / Overexpression ofHER20.992.321.8318.1817.09

[0180] As shown in Table 2, Formula 1 exhibited an activity with an IC502-fold higher than that of mobocertinib and 2.5-fold higher than that of zongertinib in the NCI-N87, a patient-derived gastric cancer cell line harboringHER2amplification / overexpression. Therefore, Formula 1 has a similar or superior level of inhibitory efficacy than the control drug against the cell line withHER2amplification and overexpression, and thus can exhibit therapeutic effects on patients withHER2amplification / overexpression andHER2-positive solid tumors accompanied therewith.

[0181] Example 3. Evaluation of anticancer activity of Formula 1 in tumor model harboringHER2mutation

[0182] Tumor models were prepared by subcutaneous or intracerebral implantation of a Ba / F3 cell line engineered to express aHER2mutant GSP or an NCI-H1781 cell line harboring aHER2mutant VC into Balb / c nude mice, and thenin vivoevaluations were performed.

[0183] Example 3-1.In vivoevaluation in subcutaneously-implanted Ba / F3HER2GSP model

[0184] Balb / c nude mice (male and female mixed, 6-7 weeks old) were purchased and acclimatized. Cultured Ba / F3HER2GSP cells were mixed with Matrigel, and 5 Х 106Ba / F3HER2GSP cells per mouse were subcutaneously implanted into the right flank of the anesthetized animals. Once tumors were formed on the skin of Balb / c nude mice, the animals were randomly divided into groups so that the average tumor volume was about 100-150 mm3, and Formula 1 or an excipient was administered for 20 days, and the tumor size was measured 2-3 times a week to calculate the volume.

[0185] Formula 1 was administered orally to the mice once daily at 25 mg / kg, 50 mg / kg, or 60 mg / kg, and as a negative control drug, 2% DMSO + 40% PEG400 + 20% Tween80 + 38% distilled water was used.

[0186] As can be seen in Fig. 1, in the subcutaneously-implanted mouse tumor model prepared by implanting a Ba / F3 cell line engineered to express aHER2mutant GSP, Formula 1 showed a statistically significant tumor growth inhibition effect starting from the lowest dose of 25 mg / kg, and showed an excellent tumor growth inhibition rate in the dose range of 50-60 mg / kg, and a tendency that the drug effect is almost saturated was observed starting from the middle dose of 50 mg / kg.

[0187] Example 3-2.In vivoevaluation in subcutaneously-implanted non-small cell lung cancer NCI-H1781HER2VC model

[0188] Balb / c nude mice (male, 6-7 weeks old) were purchased and acclimatized. Cultured NCI-H1781 HER2 VC cells were mixed with Matrigel, and 5 Х 106NCI-H1781HER2VC cells per mouse were injected into the right flank of the anesthetized animals for subcutaneous implantation. Once tumors were formed on the skin of Balb / c nude mice, the animals were randomly divided into groups so that the average tumor volume was about 100-150 mm3, and Formula 1 or an excipient was administered for 24 days, and the tumor size was measured 2-3 times a week to calculate the volume.

[0189] Formula 1 was administered orally to the mice once daily at 25 mg / kg, 50 mg / kg, or 60 mg / kg, and as a negative control drug, 2% DMSO + 40% PEG400 + 20% Tween80 + 38% distilled water was used.

[0190] As can be seen in Fig. 2, in the subcutaneously-implanted mouse tumor model prepared by implanting the NCI-H1781 cell line derived from human non-small cell lung cancer and harboring aHER2mutation VC, Formula 1 showed a statistically significant tumor growth inhibition effect starting from the lowest dose of 25 mg / kg, and showed an excellent tumor growth inhibition rate in the dose range of 25-60 mg / kg, and a tendency that the drug effect is almost saturated was observed starting from the lowest dose of 25 mg / kg.

[0191] Example 3-3. In vivo evaluation in intracerebrally-implanted non-small cell lung cancer NCI-H1781 HER2 VC model (Balb / c nude mice, male, 6-7 weeks old, oral)

[0192] Balb / c nude mice (male, 6-7 weeks old) were purchased and acclimatized. Cultured NCI-H1781 cells were diluted in PBS solution, and 5 Х 105NCI-H1781 cells per mouse were injected into the right frontal lobe of anesthetized animals for 5 minutes for intracerebral implantation. One week after intracerebral implantation, only animals with brain tumors were selected using MRI imaging, randomly divided into groups, and administered with Formula 1 or an excipient for 3 weeks. MRI measurements were performed once a week to calculate the brain tumor volume.

[0193] Formula 1 was administered orally to the mice once daily at 25 mg / kg, 50 mg / kg, or 60 mg / kg, and as a negative control drug, 2% DMSO + 40% PEG400 + 20% Tween80 + 38% distilled water was used.

[0194] As can be seen in Fig. 3, in the intracerebrally-implanted mouse tumor model prepared by implanting the NCI-H1781 cell line derived from human non-small cell lung cancer and harboring aHER2mutation VC, a tendency that the drug effect is almost saturated was observed starting from the lowest dose of 25 mg / kg.

[0195] Example 4. Evaluation of anticancer efficacy of Formula 1 in tumor model harboring HER2 mutation

[0196] A subcutaneously-implantedHER2mutant tumor model was prepared using a Ba / F3 cell line engineered to express aHER2mutant YVMA or VC and the anticancer efficacy of Formula 1 was evaluatedin vivo.

[0197] Example 4-1.In vivoevaluation in subcutaneously-implanted Ba / F3HER2YVMA model

[0198] NOD SCID mice (female, 6-8 weeks old, 17-21 g) were purchased and acclimatized. Cultured Ba / F3HER2YVMA cells (0.1 mL of a 1:1 mixture of PBS and Matrigel) at a density of 0.5 Х 106per NOD SCID mouse were subcutaneously implanted into the right flank of anesthetized NOD SCID mice. Once tumors were formed on the skin of the mice, the animals were randomly divided into groups so that the average tumor volume was constant at 134 mm3each and were orally administered with Formula 1 or an excipient (once a day) for 20 days, and the tumor size was measured three times a week and clinical symptoms were observed. Formula 1 was administered at 25 mg / kg, 50 mg / kg, or 75 mg / kg, and as a negative control drug, 2% DMSO + 40% PEG400 + 20% Tween80 + 38% distilled water was used.

[0199] As can be seen in Fig. 4, Formula 1 showed an excellent tumor growth inhibition rate starting from the 25 mg / kg dose in the subcutaneously-implanted Ba / F3 HER2 YVMA model. In addition, Formula 1 showed a complete response in all 16 subjects at the 50 mg / kg dose.

[0200] Example 4-2. Subcutaneously-implanted Ba / F3HER2VC model (NOD SCID mice, female, 6-8 weeks old, 18-22 g), tumor growth and body weight changes after 20 days of administration

[0201] NOD SCID mice (female, 6-8 weeks old, 17-21 g) were purchased and acclimatized. Cultured Ba / F3HER2VC cells (0.1 mL of a 1:1 mixture of PBS and Matrigel) at a density of 0.5 Х 106per NOD SCID mouse were subcutaneously implanted into the right flank of anesthetized NOD SCID mice. Once tumors were formed on the skin of the mice, the animals were randomly divided into groups so that the average tumor volume was constant at 123 mm3each and were orally administered with Formula 1 or an excipient for 20 days, and the tumor size was measured three times a week and clinical symptoms were observed. Formula 1 was administered at 25 mg / kg, 50 mg / kg, or 75 mg / kg, and as a negative control drug, 2% DMSO + 40% PEG400 + 20% Tween80 + 38% distilled water was used.

[0202] As can be seen in Fig. 5, Formula 1 showed an excellent tumor growth inhibition rate at a 50 mg / kg dose in the subcutaneously-implanted Ba / F3 HER2 VC model. In addition, Formula 1 showed a complete response in was observed in 11 out of 16 animals.

[0203] In summary of Example 3 and Example 4, Formula 1 exhibited excellent anticancer efficacy in a subcutaneously-implanted human non-small cell lung cancer NCI-H1781 cell line tumor model or a subcutaneously-implanted Ba / F3 cell line tumor model with YVMA, GSP, and VC, which are major subtypes of a mutation in the HER2 tyrosine kinase domain, and is thus expected to be effective in the treatment of patients with solid tumors withHER2activating mutations. Additionally, Formula 1 significantly inhibited tumor growth in the intracerebrally-implanted NCI-H1781 tumor model mimicking brain metastases, suggesting that it may be effective in preventing or treating brain metastases in patients with solid tumors with aHER2-activating mutation.

[0204] Example 5. Evaluation of anticancer efficacy of Formula 1 inHER2amplified / overexpressing tumor model

[0205] A subcutaneously-implanted wild-typeHER2amplified / overexpressing tumor model was prepared using the wild-typeHER2amplified / overexpressing NCI-N87 gastric cancer cell line, and the anticancer efficacy of Formula 1 was evaluatedin vivo.

[0206] Balb / c nude mice (male, 7-9 weeks old, 21-27 g) were purchased and acclimatized. Each mouse was subcutaneously implanted into the right flank with 1 x 107cultured NCI-N87 cells (0.2 mL of a 1:1 mixture of PBS and Matrigel) under anesthesia. Once tumors were formed on the skin of the mice, the animals were randomly divided into groups so that the average tumor volume was constant at 156 mm3each and were orally administered (once a day) with Formula 1 or an excipient for 28 days, and the tumor size was measured three times a week and clinical symptoms were observed. Formula 1 was administered at 12.5 mg / kg, 25 mg / kg, or 50 mg / kg, and as a negative control drug, 2% DMSO + 40% PEG400 + 20% Tween80 + 38% distilled water was used.

[0207] As can be seen in Fig. 6, Formula 1 showed excellent tumor growth inhibition rates at doses of 12.5 mg / kg, 25 mg / kg, and 50 mg / kg.

[0208] Formula 1 showed excellent anticancer efficacy and relatively good tolerability in a wild-typeHER2amplified / overexpressing gastric cancer cell line, and is expected to be effective in the treatment of patients with HER2 wild-typeHER2amplified / overexpressing solid tumors.

[0209] Example 6. Evaluation of anticancer efficacy of Formula 1 in subcutaneously-implanted Ba / F3EGFRSVD model

[0210] An EGFR mutant subcutaneous transplantation mouse tumor model was prepared using Ba / F3 cell line engineered to express anEGFRmutant SVD, and the anticancer efficacy of Formula 1 was evaluatedin vivo.

[0211] Balb / c nude mice (female, 6-8 weeks old, 19-22 g) were purchased and acclimatized. Cultured Ba / F3 EGFR SVD cells (0.1 mL of a 1:1 mixture of PBS and Matrigel) at a density of 0.5 Х 106per mouse were subcutaneously implanted into the right flank of anesthetized animals. Once tumors were formed on the skin of the mice, the animals were randomly divided into groups so that the average tumor volume was constant at 92 mm3each and were orally administered (once a day) with Formula 1 or an excipient for 14 days, and the tumor size was measured three times a week and clinical symptoms were observed. Formula 1 was administered at 12.5 mg / kg, 25 mg / kg, or 50 mg / kg, and as a negative control drug, 2% DMSO + 40% PEG400 + 20% Tween80 + 38% distilled water was used.

[0212] As can be seen in Fig. 7, Formula 1 showed excellent tumor growth inhibition rates at doses of 12.5 mg / kg, 25 mg / kg, and 50 mg / kg.

[0213] Formula 1 showed excellent anticancer efficacy and relatively good tolerability in a subcutaneously-implanted Ba / F3 cell line model expressing SVD, an EGFR tyrosine kinase domain mutant isoform, and is expected to be effective in the treatment of patients with solid tumors withEGFRactivating mutations.

[0214] Example 7. Results of toxicity test of 4-week repeated dose of Formula 1 (rats and beagle dogs)

[0215] Example 7-1. Oral repeated administration for 28 days and recovery toxicity at day 28 in rats

[0216] When Formula 1 was orally administered to Sprague-Dawley rats at doses of 0 mg / kg, 5 mg / kg, 10 mg / kg, or 15 mg / kg for 28 days, no mortality related to administration of the test substance was observed, and body weight loss was observed only in the male 15 mg / kg administration group. Adverse findings associated with the administration of the test substance included hemorrhage, fibrosis, and acinar cell atrophy in the pancreas. These are thought to be species-specific changes due to the rat-specific pancreatic anatomy, and thus the human risk is thought to be low. In the case of the spleen, weight loss was observed in the group administered at a concentration of 5 mg / kg or more, but complete recovery was observed in females and recovery patterns were observed in males, but weight loss was still observed in the group administered at a concentration of 10 mg / kg or more. However, no findings were observed macroscopically. Although a decrease in lymphocyte cellularity was observed histopathologically, it was at a minimal to moderate degree, all recovered, and no necrosis was observed, and thus it was determined to be a harmless finding. Therefore, the dose that causes severe toxicity in 10% of animals (severely toxic dose 10; STD10) was determined to be 15 mg / kg for both males and females.

[0217] Example 7-2. Oral repeated administration for 28 days and recovery toxicity at day 28 in beagle dogs

[0218] Formula 1 was orally administered to beagle dogs at 0 mg / kg, 0.5 mg / kg, 1 mg / kg, or 3 mg / kg for 28 days. No deaths related to the administration of the test substance occurred. However, due to weight loss in the females of 3 mg / kg administration group, a 5-day or 8-day washout period was required, and the dose was reduced to 2 mg / kg on the 20th day of administration. In the high-dose groups of both sexes, decreased activity and abnormal stools (soft stool, watery stool, mucus stool, red stool, and green stool) were observed, and body weight loss was observed in the male group administered at a dose of 3 mg / kg and the female group administered at a dose of 1 mg / kg or more. In the high-dose group of both sexes, an increase in leukocytes, neutrophils, monocytes, and eosinophils, which mainly indicate inflammatory changes, was observed, and a decrease in albumin and an increase in globulin were also observed. Adverse histopathological findings associated with the test substance administration were observed in the gallbladder (mixed cell inflammation), optic nerve (axonal degeneration), and pituitary gland (congestion / hemorrhage accompanied with pigmented macrophages) in males administered at a dose of 3 mg / kg. The no observed adverse effect level (NOAEL) was determined to be 1 mg / kg for both males and females, and the highest non-severely toxic dose (HNSTD) was determined to be 3 mg / kg for males and 1 mg / kg for females.

[0219] Example 8. Pharmacokinetics in mice, rats, and beagle dogs

[0220] Example 8-1. Pharmacokinetic evaluation according to single intravenous and oral administration in mice

[0221] Pharmacokinetics were evaluated when Formula 1 was administered intravenously as a single dose and orally at various doses to mice.

[0222] Formula 1 in a solution state dissolved in 2% DMSO + 40% PEG400 + 20% Tween 80 + 38% distilled water excipient was administered intravenously at a dose of 4 mg / kg to ICR mice, and Formula 1 in a solution state dissolved in an excipient of the same composition was administered orally at doses of 1 mg / kg, 5 mg / kg, 15 mg / kg, and 50 mg / kg. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after intravenous and oral administration. Plasma was collected by centrifugation, and the concentration of Formula 1 in plasma was quantified by LC-MS / MS analysis. The average blood concentration-time profiles of Formula 1 in mice are shown in Fig. 8, and the average pharmacokinetic parameters are shown in Table 3.

[0223] Administration RouteIntravenousOralOralOralAdministration Dose (mg / kg)451550C0or Cmax(ng / mL)130442.6245765Tmax(h)-2.673.003.17T1 / 2(h)4.35---Vdss(L / kg)5.31---CL (mL / min / kg)48.2---AUC0-last(ng·h / mL)140820812785116AUC0-inf(ng·h / mL)141422313215695

[0224] Example 8-2. Pharmacokinetic evaluation according to single intravenous and oral administration in rats

[0225] Pharmacokinetics were evaluated in rats when Formula 1 was administered intravenously as a single dose and orally at different doses.

[0226] Formula 1 in a solution state dissolved in 2% DMSO + 40% PEG400 + 20% Tween 80 + 38% distilled water excipient was administered intravenously at a dose of 2 mg / kg to SD rats, and Formula 1 in a solution state dissolved in an excipient of the same composition was administered orally at doses of 2 mg / kg and 5 mg / kg. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after intravenous and oral administration. Plasma was collected by centrifugation, and the concentration of Formula 1 in plasma was quantified by LC-MS / MS analysis. The average blood concentration-time profiles of Formula 1 in rats are shown in Fig. 9, and the average pharmacokinetic parameters are shown in Table 4.

[0227] Administration RouteIntravenousOralOralAdministration Dose (mg / kg)225C0or Cmax(ng / mL)89919.457.3Tmax(h)-4.004.00T1 / 2(h)1.98--Vdss(L / kg)3.05--CL (mL / min / kg)38.7--AUC0-last(ng·h / mL)86897.0320AUC0-inf(ng·h / mL)872112360

[0228] Example 8-3. Pharmacokinetic evaluation according to single intravenous and oral administration in beagle dogs

[0229] Pharmacokinetics of Formula 1 were evaluated in beagle dogs when administered intravenously and orally as a single dose.

[0230] Formula 1 in a solution state dissolved in 2% DMSO + 40% PEG400 + 58% distilled water excipient was administered intravenously at a dose of 1 mg / kg to beagle dogs, and Formula 1 in a solution state dissolved in an excipient of the same composition was administered orally at a dose of 2 mg / kg. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after intravenous and oral administration. Plasma was collected by centrifugation, and the concentration of Formula 1 in plasma was quantified by LC-MS / MS analysis. The average blood concentration-time profiles of Formula 1 in rats are shown in Fig. 10, and the average pharmacokinetic parameters are shown in Table 5.

[0231] Administration RouteIntravenousOralAdministration Dose (mg / kg)12C0or Cmax(ng / mL)13140.9Tmax(h)-2.67T1 / 2(h)3.65-Vdss(L / kg)10.0-CL (mL / min / kg)60.4-AUC0-last(ng·h / mL)305239AUC0-inf(ng·h / mL)307244

[0232] Example 8-4. Pharmacokinetic evaluation according to single and repeated administration in mice

[0233] Pharmacokinetics were evaluated when Formula 1 was administered orally to mice at single and repeated doses over a period of one week.

[0234] Formula 1, in a solution state dissolved in 2% DMSO + 40% PEG400 + 20% Tween 80 + 38% distilled water excipient, was orally administered to ICR mice at doses of 15 mg / kg and 50 mg / kg. Blood was collected by cross-sampling at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours before and after single and repeated oral administration. The plasma was centrifuged, and the concentration of Formula 1 in the plasma was quantified by LC-MS / MS analysis. The average blood concentration-time profiles of Formula 1 after single and repeated administration in mice are shown in Fig. 11, and the average pharmacokinetic parameters are shown in Table 6.

[0235] Administration Dose (mg / kg)15155050Number of Administration1 day7 days1 day7 daysCmax(ng / mL)209177983752Tmax(h)1.002.002.000.50AUC0-last(ng·h / mL)106088851644904AUC0-inf(ng·h / mL)115497551714909

[0236] Example 9. Pharmacokinetic modeling and simulation results for predicting human clinical dose

[0237] The international guidelines for selecting the starting dose of pharmaceuticals for cancer patients (ICH S9) recommend that the initial human starting dose be 1 / 10 of the STD10in rodent toxicity studies or 1 / 6 of the HNSTD in non-rodent studies. The conversion constants based on body area published in the FDA Guidance 2005 were used for calculations. The conversion constants from animal dose to human equivalent dose were 6.2 and 1.8 for rats and dogs, respectively. Human body weight was assumed to be 60 kg.

[0238] The calculation results are shown in Table 7. The maximum recommended starting dose (MRSD), derived from 1 / 10 of the rat STD10, was calculated to be 14.58 mg, and the MRSD for males and females, derived from 1 / 6 of the male and female dog HNSTD, were 16.23 mg and 5.41 mg, respectively.

[0239] SpeciesToxicityHEDMRSDRatsSTD10: 15 mg / kg145.8 mg14.58 mgBeagle DogsHNSTD:3 mg / kg for male1 mg / kg for female97.38 mg for male32.46 mg for female16.23 mg for male5.41 mg for female

[0240] STD10: severely toxic dose in 10 % of the animals;

[0241] HNSTD: highest non-severely toxic dose;

[0242] HED: human equivalent dose;

[0243] MRSD: maximum recommended starting dose.

[0244]

[0245] Based on the plasma concentrations of Formula 1 observed in mice, rats, and beagle dogs confirmed in Example 7 and Example 8, pharmacokinetic modeling and simulation were performed to predict the pharmacokinetic profile in humans, which is shown in Fig. 12. Pharmacokinetic modeling and simulation results showed that the steady-state simulated trough concentration (Ctrough) (0.99 ng / mL) after once-daily administration of 20 mg Formula 1 in humans was close to the simulated Ctroughof 0.91 ng / mL. Considering both MRSD and MABEL, the human starting dose was conservatively determined to be 5 mg.

[0246] As demonstrated in Examples 3 and 4, multiple antitumor efficacy tests in NOD / SCID mice or Balb / c nude mice models demonstrated maximum tumor growth inhibition at 50 mg / kg (human equivalent dose [HED] of 243 mg). Additionally, pharmacokinetic modeling and simulation predicted that the Ctrough,ss(11.51 ng / mL) at a human dose of 240 mg once daily would nearly reach the IC90(12.38 ng / mL) of thein vitrocell viability assay in Ba / F cells usingHER2-YVMA confirmed in Example 1. Considering these nonclinical results, the maximum dose in humans was determined to be 240 mg.

[0247] Therefore, the ascending dose for humans was determined to be 5 mg to 240 mg. The dose increase was planned to be about 2-fold up to a maximum of 80 mg, then 1.5-fold from 80 mg to 180 mg, and finally 1.33-fold up to 240 mg (5 mg, 10 mg, 20 mg, 35 mg, 40 mg, 50 mg, 70 mg, 80 mg, 95 mg, 120 mg, 125 mg, 165 mg, 180 mg, 220 mg, and 240 mg). Considering all of the initial dose and the human equivalent dose that showed maximum tumor growth inhibition in various preclinical models where the antitumor effect was observed, it was confirmed that 5 mg or more, particularly 5 mg to 240 mg, was an appropriate dose level to be administered.

[0248] Example 10. Efficacy of Formula 1 confirmed through phase 1 / 2 clinical trials

[0249] To demonstrate the safety, tolerability, pharmacokinetics, and antitumor activity of Formula 1 in humans, and to explore a safe and effective recommended dose (RD) for patients with locally advanced or metastatic solid tumors harboring HER2 aberrations and EGFR Exon 20 insertion mutations, this open-label, multicenter, first-in-human phase 1 / 2 clinical trial is being conducted.

[0250] Patients enrolled in the clinical trial will have locally advanced or metastatic solid tumors with a mutation in theHER2tyrosine kinase domain,HER2amplification / overexpression, or EGFR Exon 20 insertion mutation. The patient will receive Formula 1 orally once daily (QD) at a dose level corresponding to the doses confirmed in Example 9 (e.g., 5 mg, 10 mg, 20 mg, 35 mg, 40 mg, 50 mg, 70 mg, 80 mg, 95 mg, 120 mg, 125 mg, 165 mg, 180 mg, 220 mg, or 240 mg), although a switch to oral twice daily (BID) administration may be possible if a different administration frequency appears to be more beneficial based on available drug pharmacokinetic, pharmacological, patient tolerability, safety, or efficacy data from nonclinical and previous cohorts. In more frequent administration regimens, the total daily dose will be similar to or lower than the daily dose of QD therapy.

[0251] Based on the exposure (AUCssand Cmax,ss) in a simulated human weighing 60 kg and the exposure observed in STD10in rats, the safety margin of a 5 mg starting dose was estimated to be 64.14 to 73.96-fold (Table 8). Compared with the exposure in the HNSTD for male and female dogs, the safety margin of 5 mg was estimated to be 38.76 to 52.29-fold and 8.95 to 9.17-fold, respectively. The safety margin was estimated to be 1.40 to 1.60-fold the exposure (AUCssand Cmax,ss) when the simulated exposure at the highest dose in humans of 240 mg and the observed exposure at STD10inrats. Compared with the exposure at HNSTD in male and female dogs, the safety margin for 240 mg was estimated to be 0.84 to 1.13-fold and 0.20-fold, respectively.

[0252] Starting Dose of 5 mgHighest Dose of 240 mgCmax,ss(ng / mL)AUCss(hr·ng / mL)Cmax,ss(ng / mL)AUCss(hr·ng / mL)Simulated exposure for human*4.8036.64222.551681.39Safety MarginSTD10in rats (fold)†73.9664.141.601.40HNSTD in male dogs (fold)‡52.2938.761.130.84HNSTD in female dogs (fold)§9.178.950.200.20

[0253] Abbreviation: STD10, severely toxic dose in 10 % of the animals; HNSTD, highest non-severely toxic dose

[0254] * Simulation Cmax,ssand AUCssof a 60 kg human were obtained from PK modeling and simulation of Formula 1.

[0255] † The average Cmax,ssand AUCssobserved at 15 mg / kg of rat STD10were 355 ng / mL and 2,350 hr·ng / mL, respectively.

[0256] ‡ The average Cmax,ssand AUCssobserved at 3 mg / kg HNSTD in male dogs were 251 ng / mL and 1,420 hr·ng / mL, respectively.

[0257] § The average Cmax,ssand AUCssobserved at 1 mg / kg HNSTD in female dogs were 44 ng / mL and 328 hr·ng / mL, respectively.

[0258]

[0259] The current status is at the third dose level (20 mg) of Part 1. All seven subjects to date have completed safety assessments without experiencing any dose-limiting toxicities (DLTs) during the DLT evaluation period (first cycle, Week 1, Day 1 to Day 21).

Claims

1.A pharmaceutical composition for preventing or treating cancer, comprising N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:[Formula 1]2.The pharmaceutical composition of claim 1, wherein the daily dose of the active ingredient is 5 mg to 240 mg.3.The pharmaceutical composition of claim 1 or 2, wherein the pharmaceutical composition is administered once a day, twice a day, or multiple times a day.4.The pharmaceutical composition of any one of claims 1 to 3, wherein the pharmaceutical composition is administered once a day.5.The pharmaceutical composition of any one of claims 1 to 3, wherein the pharmaceutical composition is administered twice a day.6.The pharmaceutical composition of claim 1, wherein the active ingredient is orally administered once or multiple times a day in a daily dose of 5 mg to 240 mg.7.The pharmaceutical composition of any one of claims 1 to 6, wherein the pharmaceutical composition is repeatedly administered every day.8.The pharmaceutical composition of any one of claims 1 to 7, wherein the pharmaceutical composition is orally administered.9.The pharmaceutical composition of any one of claims 1 to 8, wherein the cancer is a solid tumor.10.The pharmaceutical composition of any one of claims 1 to 9,wherein the cancer is selected from the group consisting of pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, bile duct cancer, large bowel cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus and nasal cavity cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestinal cancer, meningioma, esophageal cancer, glioma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoid, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational choriocarcinoma, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cancer, acoustic schwannoma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, ovarian cancer, and thymic cancer.11.The pharmaceutical composition of any one of claims 1 to 10, wherein the cancer hasEGFRaberration and / orHER2aberration.12.The pharmaceutical composition of claim 11, wherein theEGFRaberration isEGFRamplification and / orEGFRoverexpression, and theHER2aberration isHER2amplification and / orHER2overexpression and / or a mutation in theHER2transmembrane or extracellular domain.13.The pharmaceutical composition of claim 11, wherein the mutation in theHER2transmembrane or extracellular domain is one selected from the group consisting of S310F, S310Y, R678Q, R678C, G309A, P122L, G222C, I655V, S305C, H470Q, I263T, and A293T.14.The pharmaceutical composition of any one of claims 1 to 10, wherein the cancer has anEGFRmutation and / or aHER2mutation.15.The pharmaceutical composition of claim 14, wherein theEGFRmutation is an EGFR Exon 20 insertion mutation, and / ortheHER2mutation is a mutation in theHER2tyrosine kinase domain and / or a mutation in theHER2transmembrane or extracellular domain.16.The pharmaceutical composition of claim 15, wherein the mutation in theHER2tyrosine kinase domain has an insertion mutation of exon 20 insertion and / or a point mutation.17.The pharmaceutical composition of claim 15, wherein the EGFR Exon 20 insertion mutation comprises a mutation selected from the group consisting of del19, A763_Y764insFHEA, A763_Y764insFQEA, Y764_V765insHH, A767_S768insASV, S768dupSVD, V769_D770insASV, D770_N771insNPG, D770_N771insGL, D770_N771insSVD, D770_N771insSVG, DelD770insGY, DelD770insVG, N771_P772insH, N771_P772insV, DelN771insGY, DelN771insTH, P772_H773insDNP, P772_H773insPNP, H773_V774insNPH, H773_V774insPH, H773_V774insAH, H773_V774insH, and V774_C775insHV; and / orthe mutation in theHER2tyrosine kinase domain comprises a mutation selected from the group consisting of Y772dupYVMA, DelM774insWLV, A775_G776insYVMA, A775_G776insSVMA, A775_G776insI, DelG776insVC, DelG776insLC, G778_S779insCPG, V777_G778insGSP, G778dupGSP, P780_Y781insGSP, L755S, L755P, V777L, I767M, D769H, D769N, D769Y, G776V, G776S, V777M, and V842I; and / orthe mutation in theHER2transmembrane or extracellular domain comprises one selected from the group consisting of S310F, S310Y, R678Q, R678C, G309A, P122L, G222C, I655V, S305C, H470Q, I263T, and A293T.18.The pharmaceutical composition of claim 15, wherein the cancer has an EGFR Exon 20 insertion mutation.19.The pharmaceutical composition of claim 15, wherein the cancer has a mutation in theHER2tyrosine kinase domain and / or a mutation in theHER2transmembrane or extracellular domain.20.The pharmaceutical composition of any one of claims 1 to 19, wherein the cancer has metastasized to the brain of a subject.21.The pharmaceutical composition of any one of claims 1 to 20, wherein the cancer is a recurrent or refractoryHER2-positive cancer.22.The pharmaceutical composition of claim 21, wherein the recurrent or refractoryHER2-positive cancer is recurrent or refractoryHER2-positive breast cancer, recurrent or refractoryHER2-positive gastric cancer, or recurrent or refractoryHER2-positive colorectal cancer.23.The pharmaceutical composition of claim 21 or 22, wherein the subject having recurrent or refractoryHER2-positive cancer is a subject who has received treatment with aHER2inhibitor prior to the administration of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.24.The pharmaceutical composition of any one of claims 21 to 23, wherein the subject having the recurrent or refractoryHER2-positive cancer is a subject who has acquired resistance or refractory to treatment with aHER2inhibitor.25.The pharmaceutical composition of claim 23 or 24, wherein theHER2inhibitor is aHER2tyrosine kinase inhibitor.26.The pharmaceutical composition of claim 25, wherein theHER2tyrosine kinase inhibitor is at least one selected from the group consisting of afatinib, lapatinib, neratinib, and tucatinib.27.The pharmaceutical composition of claim 23 or 24, wherein theHER2inhibitor is an anti-HER2antibody or a drug conjugate thereof.28.The pharmaceutical composition of claim 27, wherein the anti-HER2antibody or a drug conjugate thereof is at least one selected from the group consisting of trastuzumab, trastuzumab emtansine, pertuzumab, margetuximab, and trastuzumab deruxtecan.29.The pharmaceutical composition of any one of claims 21 to 23, wherein the subject having the recurrent or refractoryHER2-positive cancer has received at least two prior treatments with differentHER2inhibitors prior to the administration of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.30.The pharmaceutical composition of any one of claims 21 to 29,wherein the recurrent or refractoryHER2-positive cancer is pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, bile duct cancer, large bowel cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus and nasal cavity cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestinal cancer, meningioma, esophageal cancer, glioma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoid, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational choriocarcinoma, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cancer, acoustic schwannoma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, ovarian cancer, and thymic cancer.31.A method for preventing or treating cancer, comprising administering, to a subject, an effective amount of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof:[Formula 1]32.Use of an effective amount of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating cancer:[Formula 1]33.Use of an effective amount of N-(5-((4-((1-acetylindolin-7-yl)amino)-5-chloropyrimidin-2-yl)amino)-2-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)acrylamide represented by Formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof for preventing or treating cancer:[Formula 1]

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