Pyridonopyrimidine derivatives as RSK inhibitors and their applications

Abstract

The present invention relates to pyridonopyrimidine derivatives as RSK protein kinase inhibitors and their applications, particularly to compounds of formula I, pharmaceutical compositions containing the compounds of formula I, and the use of the compounds in the preparation of medicaments for treating RSK-related diseases or inhibiting RSK.

Description

[Technical Field]

[0001] The present invention relates to the field of medicinal chemistry, specifically, the present invention relates to novel pyridonopyrimidine derivatives, methods for their synthesis, and their application as RSK inhibitors in the preparation of drugs for tumor-related diseases. [Background technology]

[0002] Noncommunicable diseases (NCDs) are primarily comprised of four disease types: cardiovascular disease, cancer, respiratory disease, and diabetes. Currently, noncommunicable diseases are the leading cause of death in humans, with cardiovascular disease being the leading cause of human death among noncommunicable diseases, and cancer being the second leading cause of death affecting human survival. Traditional cancer treatments include surgery, radiation therapy, and chemotherapy. In recent years, with the development of science and technology, targeted therapy and immunotherapy have gradually become effective means of cancer treatment. Targeted therapy primarily involves small molecule drugs or monoclonal antibodies, which treat cancer by interfering with specific proteins to control the growth and spread of tumor cells in the body. As researchers learn more about cancer-related proteins, designing small molecule drugs targeting these proteins for cancer treatment has become increasingly promising. The Ras-MAPK signaling pathway is involved in the regulation of various types of cancer, and RSK is the downstream effector factor, whose abnormal expression and activity are associated with the development and progression of various diseases. The Ras-MAPK signaling pathway is activated by growth factors, mitogenic hormones, and neurotransmitter stimulation. Cell surface receptor activation enhances Tyr kinase autophosphorylation and creates a docking site for growth factor receptor-bound protein-2 (GRB2), connecting the receptor to a serum-free guanine nucleotide exchange factor (SOS). SOS catalyzes the binding of GTP to Ras. GTP binds to Ras and activates its effector Raf kinase. Raf phosphorylates and activates MAPK and extracellular signal-regulated kinase (MEK1 / 2). Ribosomal S6 kinase (RSK) is directly phosphorylated and activated by ERK1 / 2 and 3-phosphoinositide-dependent kinase-1 (PDK1). Activated RSK, still membrane-associated, is released into the cytoplasm or transported to the nucleus, where it mediates cell differentiation, proliferation, survival, and oncogenic transformation. The Ras signaling pathway promotes cell proliferation and protects cells from apoptosis, and is of major importance in maintaining the development, progression, and biological behavior of human tumors.

[0003] p90 ribosomal S6 protein kinase (RSK) is a member of the serine / threonine kinase family that is widely expressed in tissues. As a key downstream regulator of the Ras signaling pathway, it plays an important role in tumor development and progression. In mammals, it has four isoforms: RSK1, RSK2, RSK3, and RSK4. All four isoforms contain two kinase domains with distinct functions: an N-terminal kinase domain (NTKD) and a C-terminal kinase domain (CTKD), as well as a linker. The C-terminus contains a specific extracellular signal-regulated kinase (ERK)-binding site, and upon binding to ERK, RSK is further regulated by ERK.

[0004] Currently, there are no drugs targeting RSK in clinical trials. RSK small molecule inhibitors under development mainly include two categories: RSK2-selective inhibitors, including SL0101, CMK, etc.; and RSK pan-inhibitors, including BID-1870, FMK, LJH308, LJH685, etc.; none of these small molecule inhibitors have entered clinical research. Therefore, research and development of drugs targeting RSK has important clinical significance and application prospects. Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide pyrimidopyridone derivatives as RSK inhibitors. Another object of the present invention is to provide a pharmaceutical composition comprising the above compound. Yet another object of the present invention is to provide a use of the above compounds in the preparation of a medicament for treating RSK-related diseases or for inhibiting RSK. [Means for solving the problem]

[0006] In a first aspect, the present invention provides a compound according to formula I, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof: [ka] R 1 is hydrogen, optionally substituted C1-C 10 alkyl group, optionally substituted C3-C8 cycloalkyl group, optionally substituted C2-C6 alkenyl group, optionally substituted C3-C6 cycloalkenyl group, optionally substituted C3-C8 lactone group, optionally substituted C1-C 10 Amide group, optionally substituted C5-C 10 aryl groups, optionally substituted C3-C8 heterocyclic groups, optionally substituted C5-C 10 selected from the group consisting of aromatic heterocyclic groups;

[0007] R 2 is hydrogen, substituted C1-C 10 Alkyl groups, C3-C8 cycloalkyl groups, optionally substituted C5-C 10 aryl groups, optionally substituted C3-C8 heterocyclic groups, optionally substituted C5-C 10 selected from the group consisting of an aryl or heteroaryl group and a 5- or 6-membered heterocycle; R 3 is hydrogen, substituted C1-C 10 Alkyl groups, C3-C8 cycloalkyl groups, substituted C1-C 10 Alkylformyl group, optionally substituted C5-C 10 Arylformyl group, halogen, cyano group, optionally substituted C5-C 10 aryl groups, and optionally substituted C3-C8 heterocyclic groups.

[0008] R 4 is hydrogen, an optionally substituted C1-C6 alkyl group (e.g., trifluoromethyl group), an optionally substituted C3-C8 cycloalkyl group, an optionally substituted C1-C 10Alkylformyl group, optionally substituted C5-C 10 Arylformyl group, optionally substituted C5-C 10 It is selected from the group consisting of aryl groups, halogens (eg, fluorine), and optionally substituted C3-C8 heterocyclic groups.

[0009] In a preferred embodiment, the aryl or heteroaryl group and the 5- or 6-membered heterocycle are [ka] and 5- or 6-membered heterocycles including, but not limited to, benzo. In specific embodiments, R 1 is hydrogen, optionally substituted C1-C 10 alkyl groups, optionally substituted C3-C8 cycloalkyl groups, optionally substituted C5-C 10 aryl groups, and optionally substituted C3-C8 heterocyclic groups;

[0010] R 2 is an optionally substituted C5-C 10 aryl groups, optionally substituted C3-C8 heterocyclic groups, optionally substituted C5-C 10 It is selected from the group consisting of an aryl or heteroaryl group and a 5- or 6-membered heterocycle. R 3 is hydrogen, optionally substituted C1-C 10 alkyl groups, and optionally substituted C3-C8 cycloalkyl groups; R 4 is selected from the group consisting of hydrogen, an optionally substituted C1-C6 alkyl group (e.g., a trifluoromethyl group), an optionally substituted C3-C8 cycloalkyl group, a halogen (e.g., fluorine), and an optionally substituted C3-C8 heterocyclic group.

[0011] In a specific embodiment, the compound is a compound according to formula II: [ka] In the formula, R 1 is hydrogen, optionally substituted C1-C 10 alkyl groups, and optionally substituted C3-C8 cycloalkyl groups; R 2 is an optionally substituted C5-C 10 aryl groups, optionally substituted C3-C8 heterocyclic groups, optionally substituted C5-C 10 selected from the group consisting of an aryl or heteroaryl group and a 5- or 6-membered heterocycle;

[0012] R 4 is selected from the group consisting of hydrogen, an optionally substituted C1-C6 alkyl group (e.g., a trifluoromethyl group), and an optionally substituted C3-C8 cycloalkyl group. In specific embodiments, R 1 is an optionally substituted C3-C8 cycloalkyl group, preferably one or more halogen-substituted C3-C8 cycloalkyl groups, more preferably one or more F-substituted C3-C8 cycloalkyl groups; R 2 is an optionally substituted C5-C 10 Aryl groups (preferably phenyl groups), optionally substituted C5-C 10 is selected from the group consisting of an aryl group or a heteroaryl 5- or 6-membered heterocycle;

[0013] Said C5-C 10 Aryl group or heteroaryl 5- or 6-membered heterocycle: [ka] R 4 is selected from the group consisting of hydrogen, optionally substituted C1-C6 alkyl groups. In a specific embodiment, the compound is [ka] [ka] or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, Preferably [ka] or an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[0014] In a second aspect, the present invention provides a pharmaceutical composition comprising a compound according to the first aspect, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. In a preferred embodiment, the pharmaceutical composition is in a dosage form suitable for oral administration, including, but not limited to, tablets, solutions, suspensions, capsules, granules, and powders.

[0015] In a third aspect, the present invention provides the use of a compound according to the first aspect in the preparation of a medicament for treating or preventing an RSK protein kinase-mediated disease, or for inhibiting an RSK protein kinase, or for inhibiting one of the RSKs RSK1, RSK2, RSK3 and RSK4.

[0016] In a specific embodiment, said RSK protein kinase-mediated disease is cancer. In specific embodiments, the cancer is selected from the group consisting of esophageal cancer, renal cell carcinoma, pancreatic cancer, colon cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, endometrial cancer, head and neck squamous cell carcinoma, acute myeloid leukemia and solid tumors, or cancers and solid tumors such as breast cancer regulated by RSK1 and RSK4, ovarian cancer regulated by RSK3 and RSK4, prostate cancer regulated by RSK1 and RSK2, lung cancer regulated by RSK1, RSK2 and RSK4, head and neck squamous cell carcinoma and acute myeloid leukemia regulated by RSK2, and esophageal cancer, renal cancer, endometrial cancer, colon cancer and the like regulated by RSK4.

[0017] In a fourth aspect, the present invention provides a compound according to the first aspect for treating or preventing an RSK protein kinase mediated disease, or for inhibiting an RSK protein kinase, or for inhibiting one of the RSKs RSK1, RSK2, RSK3, RSK4. In a preferred embodiment, the RSK protein kinase mediated disease is cancer. In a preferred embodiment, the cancer is selected from the group consisting of esophageal cancer, renal cell carcinoma, pancreatic cancer, colon cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, endometrial cancer, head and neck squamous cell carcinoma, acute myeloid leukemia and solid tumors, or cancers and solid tumors such as breast cancer regulated by RSK1 and RSK4, ovarian cancer regulated by RSK3 and RSK4, prostate cancer regulated by RSK1 and RSK2, lung cancer regulated by RSK1, RSK2 and RSK4, head and neck squamous cell carcinoma and acute myeloid leukemia regulated by RSK2, and esophageal cancer, renal cancer, endometrial cancer, colon cancer and the like regulated by RSK4.

[0018] In a fifth aspect, the present invention provides a method of treating or preventing an RSK-mediated disease using a compound according to the first aspect or a pharmaceutical composition according to the second aspect. In a preferred embodiment, the RSK-mediated disease is cancer, and preferably the cancer is selected from the group consisting of breast cancer regulated by RSK1 and RSK4, ovarian cancer regulated by RSK3 and RSK4, prostate cancer regulated by RSK1 and RSK2, lung cancer regulated by RSK1, RSK2 and RSK4, head and neck squamous cell carcinoma and acute myeloid leukemia regulated by RSK2, and cancers and solid tumors such as esophageal cancer, renal cancer, endometrial cancer, and colon cancer regulated by RSK4. [Effects of the Invention]

[0019] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (e.g., in the Examples) can be combined with each other to form new or preferred technical solutions, which will not be repeated here due to space limitations. [Brief explanation of the drawings]

[0020] [Figure 1] Figure 1 shows the anti-invasive activity of compound 005 in esophageal squamous cell carcinoma. [Figure 2] 1 shows the effect of compound 005 on esophageal squamous cell carcinoma cell clonogenesis. [Figure 3] The effect of compound 005 on signal transduction pathways in esophageal squamous cell carcinoma cells is shown. DETAILED DESCRIPTION OF THE INVENTION

[0021] As a result of extensive and thorough research, the present inventors have discovered pyrimidopyridone derivatives with completely new structures, which can inhibit the activity of RSK kinase and have IC values ​​of RSK kinase inhibitory activity. 50 The values ​​reached the nM level, and the present invention was completed based on this.

[0022] Definition of Terms Definitions of some groups referred to in this specification are as follows. As used herein, the term "alkyl group" refers to a saturated branched, straight, or cycloalkyl group having a carbon chain length of 1 to 10 carbon atoms, and preferred alkyl groups include alkyl groups having a length of 1 to 5, 1 to 2, 1 to 6, 1 to 4, or 3 to 8 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and heptyl. The alkyl group may be substituted with one or more substituents, such as a halogen or halogenated alkyl group. For example, the alkyl group may be an alkyl group substituted with 1 to 4 fluorine atoms, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group.

[0023] As used herein, the term "alkenyl group" refers to a monovalent hydrocarbon group having at least one double bond, typically containing 2 to 8 carbon atoms, preferably 2 to 6 carbon atoms, and may be linear or branched. Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, isopropenyl, butenyl, isobutenyl, and hexenyl groups. As used herein, the term "ester group" generally refers to a carboxylic acid derivative having at least one ester group, which generally contains 3 to 8 carbon atoms, preferably 3 to 6 carbon atoms, and may be linear or branched. Examples of the ester group include, but are not limited to, methyl formate, ethyl formate, methyl acetate, ethyl acetate, propyl acetate, etc.

[0024] As used herein, the term "hydroxy group" refers to a branched or straight chain alcohol having a carbon chain length of 1 to 10 carbon atoms, typically containing 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms, and may be straight or branched. Examples of ester hydroxy groups include, but are not limited to, 1-hydroxy-n-butyl, 1-hydroxyisobutyl, etc. As used herein, the term "acylamino group" refers to a group having the structural formula "-R'-NH-C(O)-R", where R' can be selected from hydrogen or an alkyl group, and R can be an alkyl group, an alkenyl group, an alkynyl group, or an NR c R d an alkyl group substituted by NR c R d Alkenyl groups substituted by NR c R d substituted with an alkyl group, an alkyl group substituted with a halogen, or an alkenyl group substituted with a cyano group, wherein R c and R d can be selected from alkyl and alkenyl groups.

[0025] As used herein, the term "aryl group" refers to a monocyclic, bicyclic, or tricyclic aromatic group containing 6 to 14 carbon atoms, including phenyl, naphthyl, phenanthrenyl, anthracenyl, indenyl, fluorenyl, tetrahydronaphthyl, indanyl, etc. An aryl group does not include halogen, C 1-4 Aldehyde group, C 1-6 Alkyl groups, cyano groups, nitro groups, amino groups, amido groups, hydroxy groups, hydroxymethyl groups, alkyl groups substituted with halogen (e.g., trifluoromethyl groups), alkoxy groups substituted with halogen (e.g., trifluoromethoxy groups), carboxy groups, C 1-4 Alkoxy group, ethoxycarbonyl group, N(CH3) and C 1-4 It can be optionally substituted with 1 to 5 (eg, 1, 2, 3, 4, or 5) substituents selected from acyl groups, heterocyclic groups, heteroaryl groups, and the like.

[0026] As used herein, the term "heterocyclic group" includes, but is not limited to, 5- or 6-membered heterocyclic groups containing 1 to 3 heteroatoms selected from O, S, and N, including, but not limited to, furanyl, thienyl, pyrrolyl, pyrrolidinyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, pyranyl, pyridinyl, pyrimidinyl, pyrazinyl, piperidinyl, and morpholinyl groups. As used herein, the term "aromatic heterocyclic group" refers to a group containing 5 to 14 ring atoms and having 6, 10, or 14 electrons shared among the ring system. The ring atoms are carbon atoms and 1 to 3 heteroatoms selected from oxygen, nitrogen, and sulfur. Useful aromatic heterocyclic groups include piperazinyl, morpholinyl, piperidinyl, pyrrolidinyl, thienyl, furanyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, and pyridinyl groups (including, but not limited to, pyrimidinyl groups). Aromatic heterocyclic groups may contain halogens, C 1-4 Aldehyde group, C 1-6 A straight-chain or branched alkyl group, a cyano group, a nitro group, an amino group, a hydroxy group, a hydroxymethyl group, an alkyl group substituted with a halogen (e.g., a trifluoromethyl group), an alkoxy group substituted with a halogen (e.g., a trifluoromethoxy group), a carboxy group, C 1-4 Alkoxy group, ethoxycarbonyl group, N(CH3) and C 1-4 It can be substituted with 1 to 5 (for example, 1, 2, 3, 4, or 5) substituents arbitrarily selected from acyl groups.

[0027] As used herein, the term "alkoxy group" refers to an oxy group substituted with an alkyl group. Preferred alkoxy groups are those having a length of 1 to 6 carbon atoms, more preferably those having a length of 1 to 3 carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, and propoxy groups. The alkoxy group may be substituted with one or more substituents, such as a halogen or halogenated alkyl group. For example, the alkoxy group may be an alkyl group substituted with 1 to 4 fluorine atoms, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group.

[0028] As used herein, "halogen" refers to fluorine, chlorine, bromine or iodine. Based on the teachings of the present invention and common knowledge in the art, those skilled in the art will understand that each group in the compound of the present invention can be further substituted to obtain a derivative having the same or similar activity as the compound specifically disclosed in the present invention. Each group in the compound of the present invention can be substituted with various substituents conventional in the art, as long as the substitution does not violate the rules of chemical synthesis or valence. The term "substituted" as used herein refers to the replacement of one or more hydrogen atoms on a specific group with a specific substituent. The specific substituent may be the above-mentioned substituent, a specific substituent in each example, or a common substituent in the art. Therefore, in the present invention, the substituents in the general formula may each independently correspond to the corresponding group in the specific compound in the examples. That is, the present invention includes not only the combination of each substituent in the above general formula, but also the combination of some of the substituents shown in the general formula with other specific substituents in the examples. It is not difficult to prepare compounds having such combinations of substituents and determine whether the resulting compounds are active based on conventional techniques in the art. In other words, based on the teachings of the present invention, those skilled in the art can synthesize various compounds within the scope of protection of the present invention. These compounds are not limited to the specific compounds partially disclosed in the examples herein. The compounds of the present invention include the specific compounds partially disclosed in the examples, or various compounds formed by specific substituents at specific substitution positions in these specific compounds and substituents at other substitution positions in the general formula, which will not be repeated here due to space limitations.

[0029] As used herein, "optionally substituted" means that the substituent it modifies is selected from the group consisting of halogen, C 1-4 Aldehyde group, C 1-6 A straight-chain or branched alkyl group, a cyano group, a nitro group, an amino group, a hydroxy group, a hydroxymethyl group, an alkyl group substituted with a halogen (e.g., a trifluoromethyl group), an alkoxy group substituted with a halogen (e.g., a trifluoromethoxy group), a carboxy group, C 1-4 Alkoxy group, ethoxycarbonyl group, N(CH3) and C 1-4 It refers to being substituted with 1 to 5 (for example, 1, 2, 3, 4, or 5) substituents arbitrarily selected from acyl groups.

[0030] Compounds of the Invention The present inventors synthesized candidate compounds with RSK inhibitory activity. They optimized the structures of the resulting candidate compounds and designed, synthesized, and structurally characterized a series of pyrimidopyridone compounds that have not been reported in the literature. They tested the activity of these compounds at the molecular level and obtained compounds capable of inhibiting RSK kinase activity. The compound of the present invention is a compound of formula I or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 1 , R 2 , R 3 , R 4 is as described above. Furthermore, the compound of the present invention may be a compound represented by formula II or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 1 , R 2 , R 4 is as described above.

[0031] In a specific embodiment, the present invention provides a series of pyrimidopyridone compounds whose structures have not been reported in the literature, and specific compounds are as follows: [ka] [ka]

[0032] Those skilled in the art will appreciate that the compounds of the present invention further include all pharmaceutically acceptable isotopically labeled compounds, where one or more atoms are replaced by atoms having the same atomic number, but the atomic mass or mass number is different from the atomic mass or mass number normally occurring in nature. Suitable isotopes for inclusion in the compounds of the present invention include, for example, 2 H and 3 Isotopes of hydrogen such as H, e.g. 11 C.13 C and 14 Isotopes of carbon such as C, e.g. 13 N and 15 Isotopes of nitrogen such as N, e.g. 15 O. 17 O and 18 It includes isotopes of oxygen such as 2O. For example, deuterium, i.e. 2 Substitution with heavier isotopes such as H can offer certain therapeutic advantages, such as greater metabolic stability, e.g., in vivo half-life, increased or decreased dosage requirements, and therefore may be preferable in certain cases. Based on the teachings of the present invention, those skilled in the art will understand that the compound of the present invention is a compound with pharmacological activity and can be used as a drug.As a pharmaceutically active compound, the compound of the present invention should naturally have various inherent properties of a drug, such as drug potential, bioavailability, low toxicity, side effects, etc.Based on the teachings of the present invention, those skilled in the art can use various well-known technical means to obtain the compound of the present invention and test the various properties described above.In other words, based on the teachings of the present invention, those skilled in the art can repeat or detect the present invention.

[0033] Based on the compounds of the present invention, the present invention provides pharmaceutical compositions, which comprise a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. Examples of pharmaceutically acceptable salts of the compounds of the present invention include, but are not limited to, inorganic and organic acid salts such as, for example, hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate, fumarate, mandelate, and oxalate salts, as well as inorganic and organic base salts formed with bases such as, for example, sodium hydroxide, tris(hydroxymethyl)aminomethane (TRIS, tromethamine), and N-methylglucamine.

[0034] The pharmaceutical compositions of the present invention can be prepared in dosage forms suitable for various routes of administration, including, but not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, intrathecal, intracranial, intranasal, or external administration for the treatment of tumors and other diseases. The dosage is an amount effective to ameliorate or eliminate one or more symptoms. For the treatment of a specific disease, an effective amount is an amount sufficient to improve or alleviate in a specific manner symptoms associated with the disease. Such an amount can be administered as a single dose or according to an effective treatment regimen. While a dosage may cure a disease, it is usually administered to improve symptoms of the disease. Repeated administration is usually required to achieve the desired improvement in symptoms. The dosage is determined based on the patient's age, health, weight, type of concurrent treatment, frequency of treatment, and the desired therapeutic effect.

[0035] The pharmaceutical preparations of the present invention can be administered to any mammalian species, most importantly humans, so long as the therapeutic effects of the compounds of the present invention are obtained. The compounds of the present invention or pharmaceutical compositions thereof can be used to treat various RSK protein kinase-mediated diseases. In this specification, the RSK protein kinase-mediated diseases are various cancers. The cancers include, but are not limited to, breast cancer regulated by RSK1 and RSK4, ovarian cancer regulated by RSK3 and RSK4, prostate cancer regulated by RSK1 and RSK2, lung cancer regulated by RSK1, RSK2, and RSK4, head and neck squamous cell carcinoma and acute myeloid leukemia regulated by RSK2, and cancers and solid tumors such as esophageal cancer, kidney cancer, endometrial cancer, and colon cancer regulated by RSK4.

[0036] The pharmaceutical preparations of the present invention can be prepared by known methods, such as conventional mixing, granulation, tableting, dissolving, or freeze-drying processes. When preparing oral preparations, solid excipients and active compounds can be combined, and the mixture can be optionally ground. If necessary, after adding appropriate excipients, the granulated mixture is processed to obtain tablet or lozenge cores. Suitable excipients are, in particular, fillers, such as sugars such as lactose or sucrose, mannitol or sorbitol, cellulose preparations or calcium phosphate salts, such as tricalcium phosphate or calcium hydrogen phosphate, and binders, such as starch pastes containing corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone. If necessary, disintegrants can be added, such as the above-mentioned starches, carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, or alginic acid or its salts, such as sodium alginate. Excipients are, in particular, flow regulators and lubricants, such as silica, talc, stearates, such as magnesium calcium stearate, stearic acid, or polyethylene glycol. If necessary, the core of the lozenge can be coated with a suitable coating that is resistant to gastric juices. Therefore, concentrated sugar solutions can be used. The solution may contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, a lacquer solution, and a suitable organic solvent or solvent mixture. To prepare a coating that is resistant to gastric juices, a suitable cellulose solution such as cellulose acetate phthalate or hydroxypropylmethylcellulose phthalate may be used. Dyes or pigments may be added to the coating of the tablet or lozenge core, for example, to identify or characterize the dosage combination of the active ingredients.

[0037] Based on the above compounds and pharmaceutical compositions, the present invention further provides a method for treating an RSK protein kinase-mediated disease, the method comprising administering to a subject in need thereof a compound or pharmaceutical composition of the present invention. The administration method includes, but is not limited to, various administration methods well known in the art, and can be determined based on the actual condition of the patient, including, but not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, intrathecal, intracranial, intranasal or topical administration routes. The present invention also includes the use of a compound of the present invention in the preparation of a medicament for preventing or treating an RSK-mediated disease or for inhibiting RSK4 activity.

[0038] Advantages of this invention: 1. The compounds provided by the present invention are pyridopyrimidone compounds with a completely new structure. 2. The compounds provided by the present invention have excellent inhibitory activity against RSK protein kinase. 3. The compounds provided by the present invention lay the foundation for the development of drugs that can inhibit target RSK, and have great industrialization and commercialization prospects, as well as market value, and significant economic benefits. The technical solutions of the present invention will be further described below in conjunction with specific examples, but the following examples do not constitute a limitation of the present invention, and all various application methods adopted according to the principles and technical means of the present invention all belong to the scope of the present invention. In the following examples, experimental methods without specific conditions are usually in accordance with conventional conditions or conditions suggested by manufacturers. Unless otherwise specified, percentages and parts are calculated by weight.

[0039] Materials and Methods The pyrimidopyridone compounds of the present invention can be synthesized as follows.

[0040] Example 1 2-((1H-indazol-5-yl)amino)-8-cyclopentyl-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (001) [ka] [ka]

[0041] Step 1. 5-Bromo-2-chloro-N-cyclopentylpyrimidin-4-amine (1) [ka] 5-Bromo-2,4-dichloropyrimidine (3.2 g, 14.2 mmol) and cyclopentylamine (1.2 g, 14.2 mmol) were added sequentially to a 100 mL single-neck round-bottom flask, followed by 15 mL of acetonitrile as solvent. After cooling to 0 °C in an ice-salt bath, K2CO3 (3.9 g, 28.4 mmol) was added and stirred in the ice bath for 10 minutes. The mixture was then warmed to room temperature and stirred for 10 hours. The reaction progress was monitored by thin-layer chromatography (TLC). After completion of the reaction, 100 mL of water was added, followed by extraction with ethyl acetate. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield a white oily liquid. The product was separated and purified by high-performance silica gel column chromatography (petroleum ether / ethyl acetate = 30:1) to obtain 2.6 g of the corresponding white solid, 5-bromo-2-chloro-N-cyclopentylpyrimidin-4-amine, in a 66% yield.

[0042] 1 H NMR(400MHz,DMSO-d6)δ 9.00(s,1H),8.61(d,J=7.6Hz,1H),4.53-4.44(m,1H),2.00-1.95(m,2H),1.7 5-1.69(m,2H),1.67-1.62(m,2H),1.60-1.56(m,2H).LC-MS:m / z:275.3(M+H) + .

[0043] Step 2. 2-Chloro-8-cyclopentyl-5-methylpyridine[2,3-d]pyrimidin-7(8H)-one (2) [ka]

[0044] The intermediate 5-bromo-2-chloro-N-cyclopentylpyrimidin-4-amine (1.5 g, 5.5 mmol), crotonic acid (2.3 g, 27.5 mmol), bis(benzonitrile)palladium chloride (230 mg, 0.6 mmol), tris(o-methylphenyl)phosphine (183 mg, 0.6 mmol), N,N-diisopropylethylamine (7.2 g, 55 mmol), and 6 mL of n-butanol were added to a 50 mL two-neck round-bottom flask and purged with nitrogen three times. The mixture was incubated at 95 °C for 4 hours and then monitored by TLC. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by high-performance silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain a brown-red oily liquid. 10 mL of acetic anhydride was added to the brown-red oily liquid and the mixture was incubated at 130 °C for 1 hour. The reaction progress was monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by high-performance silica gel column chromatography (dichloromethane / methanol = 300:1) to obtain 434 mg of a pale yellow solid, intermediate 2, in a yield of 30%.

[0045] 1 H NMR(400MHz,DMSO-d6)δ 8.29(s,1H),7.72(s,1H),4.63-4.49(m,1H),2.51(s,3H),2.44-2.19(m,2H),2.17-2.10(m,3H),2.00-1.86(m,3H).LC-MS:m / z:264.1(M+H) + .

[0046] Step 3. 2-((1H-indazol-5-yl)amino)-8-cyclopentyl-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (001) [ka]

[0047] A 100 mL single-neck round-bottom flask was charged with 2-chloro-8-cyclopentyl-5-methylpyridine[2,3-d]pyrimidin-7(8H)-one (400 mg, 1.5 mmol), 5-aminoindazole (202 mg, 1.5 mmol), p-toluenesulfonic acid (261 mg, 1.5 mmol), and 4 mL of n-butanol. The mixture was then reacted at 95 °C for 12 hours, and the reaction progress was monitored by TLC. After completion of the reaction, 100 mL of saturated aqueous NaCl solution was added, and the mixture was extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, and the solvent was removed by concentration under reduced pressure. Separation by column chromatography (dichloromethane / methanol = 30:1) afforded 145 mg of the corresponding compound 001 as a pale yellow solid in a 27% yield.

[0048] 1 H NMR(400MHz,DMSO-d6)δ 13.00(s,1H),9.94(s,1H),8.81(s,1H),8.12(s,1H),8.00(s,1H),7.53(q,J=8.7Hz,2H),6.18( s, 1H), 5.84-5.80 (m, 1H), 2.37 (s, 3H), 2.25-2.20 (m, 2H), 1.88-1.69 (m, 4H), 1.56-1.50 (m, 2H). 13 C NMR(151MHz,DMSO-d6)δ 163.01,159.72,157.27,156.04,145.83,137.32,133.56,132.98,123.33,122.52,1 17.61,111.20,110.42,107.20,53.01,27.95,25.45,17.15.HRMS(ESI)(m / z):[M+H] + calcd for C 20 H 20 N6O,361.1780;found 361.1779.

[0049] N-(8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyridine[2,3-d]pyrimidin-2-yl)-N-(3,5-difluoro-4-hydroxyphenyl)acetamide (002) [ka]

[0050] The synthesis method was the same as in Example 1. 8-Cyclopentyl-2-((3,5-difluoro-4-hydroxyphenyl)amino)-5-methylpyridine[2,3-d]pyrimidin-7(8H)-one (003) [ka]

[0051] The synthesis method was the same as in Example 1. Light yellow solid, 35% yield. 1 H NMR(400MHz,DMSO-d6)δ 10.01(s,1H),9.77(s,1H),8.83(s,1H),7.44(d,J=10.0Hz,2H),6.21(s,1H),5.89-5.69(m,1 H),2.37(s,3H),2.25-2.20(m,2H),1.93-1.80(m,2H),1.83-1.69(m,2H),1.67-1.53(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 162.88,158.92,157.23,155.94,153.34,153.28,151.76,151.70,145.77,131.70,129.02,1 28.92,118.33,107.66,104.04,103.99,103.86,27.93,25.45,17.13.HRMS(ESI)(m / z):[M+H] + calcd for C 19 H 18 F2N4O2,373.1476;found 373.1475.

[0052] 2-((1H-benzo[d][1,2,3]triazol-5-yl)amino)-8-cyclopentyl-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (004) [ka]

[0053] The synthesis method was the same as in Example 1. Light yellow solid, 35% yield. 1 H NMR(400MHz,DMSO-d6)δ 15.56(s,1H),10.26(s,1H),8.89(s,1H),8.33(s,1H),7.95(s,1H),7.63(s,1H),6.24(s,1H),6.00-5 .76(m,1H),2.40(s,3H),2.25-2.20(m,2H),1.95-1.90(m,2H),1.82-1.78(m,2H),1.70-1.56(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 163.11,159.67,157.56,155.76,146.56,137.32,133.60,132.85,123.33,122.36, 117.41,111.08,110.48,60.83,35.16,19.11,17.21,14.33.HRMS(ESI)(m / z):[M+H] + calcd for C 19 H 19 N7O,362.1730;found 362.1731.

[0054] 2-((1H-indazol-5-yl)amino)-8-(3,3-difluorocyclopentyl)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (005) [ka] [ka]

[0055] Step 1. 5-Bromo-2-chloro-N-(3,3-difluorocyclopentyl)pyrimidin-4-amine (3) [ka] A 100 mL single-neck round-bottom flask was charged with 5-bromo-2,4-dichloropyrimidine (2.0 g, 8.9 mmol), 3,3-difluorocyclopentylamine hydrochloride (1.4 g, 8.9 mmol), and then 15 mL of acetonitrile. After cooling to 0°C in an ice-salt bath, K2CO3 (2.5 g, 17.8 mmol) was added and stirred for 10 minutes in the ice bath. The mixture was then warmed to room temperature and stirred for 10 hours. The reaction progress was monitored by TLC. After completion of the reaction, 100 mL of water was added, and the mixture was extracted three times with ethyl acetate. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white oily liquid. After separation and purification by high-performance silica gel column chromatography (petroleum ether / ethyl acetate=30:1), 1.9 g of the corresponding white solid 5-bromo-2-chloro-N-(3,3-difluorocyclopentyl)pyrimidin-4-amine was obtained, with a yield of 70%.

[0056] Step 2. 2-Chloro-8-(3,3-difluorocyclopentyl)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (4) [ka] A 50 mL two-neck round-bottom flask was charged with 5-bromo-2-chloro-N-cyclopentylpyrimidin-4-amine (1.5 g, 5 mmol), crotonic acid (2.2 g, 25 mmol), bis(benzonitrile)palladium chloride (191 mg, 0.5 mmol), tris(o-methylphenyl)phosphine (152 mg, 0.5 mmol), N,N-diisopropylethylamine (6.5 g, 50 mmol), and 6 mL of n-butanol, followed by three nitrogen purges. The reaction mixture was incubated at 95 °C for 4 hours and monitored by TLC. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by high-performance silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain a brown-red oily liquid. 10 mL of acetic anhydride was added to the brown-red oily liquid, and the mixture was incubated at 130 °C for 1 hour. The reaction progress was monitored by TLC. After the reaction was completed, the solvent was removed by direct concentration under reduced pressure, and the residue was separated and purified by high-performance silica gel column chromatography (dichloromethane / methanol = 300:1) to obtain 449 mg of pale yellow solid 2-chloro-8-(3,3-difluorocyclopentyl)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one, with a yield of 30%.

[0057] 1 H NMR(400MHz,DMSO-d6)δ 9.07(s,1H),6.64(s,1H),5.99-5.83(m,1H),2.96-2.85(m,1H),2.65(m,1H) ,2.46(s,3H),2.43-2.31(m,2H),2.28-2.08(m,2H).LC-MS:m / z:300.2(M+H) + .

[0058] Step 3. 2-((1H-indazol-5-yl)amino)-8-(3,3-difluorocyclopentyl)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (005) [ka] A 100 mL single-neck round-bottom flask was charged with 2-chloro-8-(3,3-difluorocyclopentyl)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (400 mg, 1.3 mmol), 5-aminoindazole (178 mg, 1.3 mmol), p-toluenesulfonic acid (224 mg, 1.3 mmol), and 4 mL of n-butanol. The reaction mixture was incubated at 95 °C for 12 hours, and the reaction progress was monitored by TLC. After completion of the reaction, 100 mL of saturated aqueous NaCl solution was added, followed by extraction with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, and the solvent was removed by concentration under reduced pressure. Separation by column chromatography (dichloromethane / methanol = 30:1) afforded 180 mg of 005 as a yellow solid in a 35% yield.

[0059] 1 H NMR(600MHz,DMSO-d6)δ(ppm):13.00(s,1H),10.05(s,1H),8.85(s,1H),8.17-8.1 2(m,1H),7.97(s,1H),7.56(dd,J=8.9,1.9Hz,1H),7.50(d,J=8.9Hz,1H),6.21(d,J =1.4Hz,1H),6.11(s,1H),3.03(dq,J=27.6,11.9Hz,1H),2.49-2.41(m,2H),2.39( d,J=1.2Hz,3H),2.28(ddt,J=19.0,13.2,6.5Hz,1H),2.12-1.96(m,2H).LC-MS:m / z calc.for C 20 H 18 F2N6O[M+H] + :397.15,found 397.20.

[0060] 2-((1H-indazol-5-yl)amino)-5-methyl-8-(pyrrolidin-3-amino)pyrido[2,3-d]pyrimidin-7(8H)-one (006) [ka] [ka]

[0061] Step 1. 3-(5-Bromo-2-chloropyrimidin-4-yl)amino)pyrrolidine-1-carboxylic acid t-butyl ester (5) [ka] A 100 mL single-neck round-bottom flask was charged with 5-bromo-2,4-dichloropyrimidine (2.5 g, 11.1 mmol), 3-aminopyrrolidine-1-carboxylic acid tert-butyl ester (2.1 g, 11.1 mmol), and then 15 mL of acetonitrile as solvent. After cooling to 0 °C in an ice-salt bath, K2CO3 (3.1 g, 22.2 mmol) was added and stirred in the ice bath for 10 minutes. The mixture was then warmed to room temperature and stirred for 10 hours. The reaction progress was monitored by TLC. After completion of the reaction, 100 mL of water was added, and the mixture was extracted three times with ethyl acetate. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white oily liquid. Separation and purification by high-performance silica gel column chromatography (petroleum ether / ethyl acetate=30:1) gave the corresponding white solid 3-(5-bromo-2-chloropyrimidin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester 2.1 g, yield 50%.

[0062] 1 H NMR(400MHz,DMSO-d6)δ 8.30(s,1H),7.64(d,J=6.9Hz,1H),4.74-4.34(m,1H),3.58(dd,J=10.8,7.3H z,1H),3.46-3.36(m,1H),3.31-3.09(m,2H),2.19-1.85(m,2H),1.41(s,9H).

[0063] Step 2. 3-(2-chloro-5-methyl-7-oxopyridine[2,3-d]pyrimidin-8(7H)-yl)pyrrolidine-1-carboxylic acid t-butyl ester (6) [ka] A 50 mL two-neck round-bottom flask was charged with 5-bromo-2-chloro-N-cyclopentylpyrimidin-4-amine (1.5 g, 4.0 mmol), crotonic acid (1.7 g, 20 mmol), bis(benzonitrile)palladium chloride (153 mg, 0.4 mmol), tris(o-methylphenyl)phosphine (122 mg, 0.4 mmol), N,N-diisopropylethylamine (5.1 g, 40 mmol), and 6 mL of n-butanol, followed by three nitrogen purges. The mixture was incubated at 95 °C for 4 hours and monitored by TLC. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by high-performance silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain a brown-red oily liquid. 10 mL of acetic anhydride was added to the brown-red oily liquid and the mixture was incubated at 130 °C for 1 hour. The reaction progress was monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by high-performance silica gel column chromatography (dichloromethane / methanol = 300:1) to obtain 145 mg of a pale yellow solid, 3-(2-chloro-5-methyl-7-oxopyridine[2,3-d]pyrimidin-8(7H)-yl)pyrrolidine-1-carboxylic acid tert-butyl ester, in a yield of 10%.

[0064] 1 H NMR(400MHz,DMSO-d6)δ 9.06(s,1H),6.65(s,1H),6.13-5.79(m,1H),3.84-3.50(m,3H),2.46(s,3H),2.22-2.20(m,1H),1.87-1.78(m,1H),1.41(d,J=13.0Hz,9H).

[0065] Step 3. 2-((1H-indazol-5-yl)amino)-5-methyl-8-(pyrrolidin-3-amino)pyrido[2,3-d]pyrimidin-7(8H)-one (006) [ka]

[0066] A 100 mL single-neck round-bottom flask was charged with 3-(2-chloro-5-methyl-7-oxopyridine[2,3-d]pyrimidin-8(7H)-yl)pyrrolidine-1-carboxylic acid t-butyl ester (200 mg, 0.6 mmol), 5-aminoindazole (80 mg, 0.6 mmol), p-toluenesulfonic acid (103 mg, 0.6 mmol), and 4 mL of n-butanol. The mixture was reacted at 95 °C for 12 hours, and the reaction progress was monitored by TLC. After completion of the reaction, 100 mL of saturated aqueous NaCl solution was added, and the mixture was extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, and the solvent was removed by concentration under reduced pressure. Separation by column chromatography (dichloromethane / methanol = 30:1) afforded 43 mg of 006 as a pale yellow solid in a 20% yield.

[0067] 1 H NMR(400MHz,DMSO-d6)δ 13.05(s,1H),9.95(s,1H),8.82(s,1H),8.10(s,1H),8.04(s,1H),7.61-7.43(m,2H),6.20(s,1H),5.96-5.90(m,1H),3.17(s ,1H),3.09(dd,J=11.4,5.9Hz,2H),2.90-2.80(m,1H),2.69-2.63(m,1H),2.38(s,3H),2.11-2.06(m,1H),2.03-1.94(m,1H). 13 C NMR(151MHz,DMSO-d6)δ 163.09,163.07,159.71,157.37,155.96,146.05,146.02,132.90,128.50,128.48,125.97,125.95,122.4 5,122.42,117.43,111.26,49.89,48.39,30.24,29.49,27.02,21.25,17.18,0.58.HRMS(ESI)(m / z):[M+H] + calcd for C 19 H 19 N7O,362.1729;found 362.1727.

[0068] 8-Cyclopentyl-5-methyl-2-((2-oxindol-5-yl)amino)pyridine[2,3-d]pyrimidin-7(8H)-one (007) [ka] The synthesis method was the same as in Example 1.

[0069] Light yellow solid, 39% yield. 1 H NMR(400MHz,DMSO-d6)δ 10.33(s,1H),9.83(s,1H),8.74(s,1H),7.61(s,1H),7.43(d,J=8.4Hz,1H),6.75(d,J=8.0Hz,1H),6.16(s,1H),5 .81(s,1H),3.43(s,2H),2.30(s,3H),2.34-2.14(m,2H),1.92-1.90(m,2H),1.77-1.64(m,2H),1.65-1.51(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 176.69,162.98,159.47,157.23,156.02,145.82,139.52,134.08,126.41,120. 27,118.14,117.62,109.25,36.53,27.98,25.54,17.13.HRMS(ESI)(m / z):[M+H] + calcd for C 21 H 21 N5O2,376.1773;found 376.1772.

[0070] 6-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyridine[2,3-d]pyrimidin-2yl)amino)benzo[d]oxazol-2(3H)-one (008) [ka] The synthesis method was the same as in Example 1. Light yellow solid, 39% yield. 1H NMR(400MHz,DMSO-d6)δ 9.91(s,1H),8.80(s,1H),7.48(s,1H),7.27(dd,J=8.6,1.9Hz,1H),7.17(d, J=8.6Hz,1H),6.19(s,1H),5.87-5.75(m,1H),2.37(s,3H),2.18-2.00(m,2H ),1.88-1.80(m,2H),1.80-1.70(m,2H),1.63-1.52(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 162.92,159.35,157.21,156.02,156.59,145.78,139.99,135.93,130.41,117.95,1 13.52,109.14,107.24,103.09,52.94,28.17,25.72,17.15.HRMS(ESI)(m / z):[M+H] + calcd for C 20 H 19 N5O3,378.1566;found 378.1565.

[0071] 8-Cyclopentyl-2-((7-fluoro-1H-indazol-5-yl)amino)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (009) [ka] The synthesis method was the same as in Example 1. Light yellow solid, 39% yield.

[0072] 1 H NMR(400MHz,DMSO-d6)δ 13.56(s,1H),10.07(s,1H),8.84(s,1H),8.12(s,1H),7.91(s,1H),7.59(d,J=13.2Hz,1H),6.20(s,1H),5 .97-5.73(m,1H),2.38(s,3H),2.26-2.20(m,2H),1.88-1.80(m,2H),1.78-1.70(m,3H),1.58-1.52(m,2H). 13C NMR(151MHz,DMSO-d6)δ 162.97,159.45,157.30,156.00,148.01,146.39,145.83,134.40,133.63,126.72,1 26.64,117.98,107.51,106.50,53.16,27.97,25.44,17.15.HRMS(ESI)(m / z):[M+H] + calcd for C 20 H 19 FN6O,379.1683;found 379.1681.

[0073] 2-((1H-indazol-5-yl)amino)-6-bromo-8-cyclopentyl-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (011) [ka] The synthesis method was the same as in Example 1.

[0074] 1 H NMR(400MHz,DMSO-d6)δ 13.02(s,1H),10.10(s,1H),8.99(s,1H),8.08(s,1H),8.00(s,1H),7.54(q,J=8.7Hz,2H), 5.98-5.55(m,1H),2.56(s,3H),2.120-2.12(m,2H),1.88-1.79(m,4H),1.58-1.51(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 159.57,158.57,158.14,154.67,145.20,142.9,137.37,133.61,132.79,123.32,1 22.42,114.8,111.26,110.47,60.01,28.01,25.60,18.36.HRMS(ESI)(m / z):[M+H] + calcd for C 20 H 19 BrN6O,439.0882;found 439.0884.

[0075] 8-Cyclopentyl-2-(3-fluoro-1H-indazol-5-yl)amino)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (012) [ka] The synthesis method was the same as in Example 1.

[0076] 2-((1H-pyrazole[3,4-b]pyridin-5-yl)amino)-8-cyclopentyl-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (013) [ka] The synthesis method was the same as in Example 1. Light yellow solid, 39% yield. 1 H NMR(400MHz,DMSO-d6)δ 13.61(s,1H),10.06(s,1H),8.83(s,1H),8.71(s,1H),8.46(s,1H),8.10(s,1H),6.21(s,1 H),5.88-5.70(m,1H),2.38(s,3H),2.332.19(m,2H),1.79-1.69(m,4H),1.59-1.50(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 162.97,159.84,157.40,156.02,149.22,145.85,145.31,133.27,130.50,120.97,1 17.89,114.47,108.60,107.68,52.94,32.01,27.96,17.17.HRMS(ESI)(m / z):[M+H] + calcd for C 19 H 19 N7O,362.1730;found 362.1730.

[0077] 6-Bromo-8-cyclopentyl-5-methyl-2-(2-oxoindole-5-yl)amino)pyridine[2,3-d]pyrimidin-7(8H)-one (014) [ka]

[0078] The synthesis method was the same as in Example 1. 1 H NMR(400MHz,DMSO-d6)δ 10.33(s,1H),9.98(s,1H),8.93(s,1H),7.61(s,1H),7.43(d,J=7.7Hz,1H),6.78(d,J=8.3Hz,1H),5.99-5.89 (m,1H),3.48(s,2H),2.55(s,3H),2.25-2.10(m,2H),1.99-1.89(m,2H),1.84-1.74(m,2H),1.65-1.59(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 176.69,159.30,158.51,158.08,154.65,145.16,139.66,133.87,130.12,126.45,120. 26,118.09,114.93,109.27,60.83,36.52,28.03,25.69,18.33.HRMS(ESI)(m / z):[M+H] + calcd for C 21 H 20 BrN5O2,454.0876;found 454.0879.

[0079] 8-(3,3-Difluorocyclopentyl)-5-methyl-2-((2-oxindol-5-yl)amino)pyridine[2,3-d]pyrimidin-7(8H)-one (015) [ka] Light yellow solid, 39% yield. 1H NMR(400MHz,DMSO-d6)δ 10.33(s,1H),9.93(s,1H),8.81(s,1H),7.60(s,1H),7.42(d,J=8.2Hz,1H),6.77(d,J=8.0Hz,1H),6.17(d,J=16.0Hz,1H),6.15 -6.00(m,1H),3.46(s,2H),3.01(td,J=24.0,11.9Hz,1H),2.49-2.40(m,2H),2.37(s,3H),2.29-2.19(m,1H),2.19-2.00(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 176.69,163.09,159.47,157.52,155.72,146.53,139.67,133.88,126.46,120.40,118 .25,117.35,109.27,60.83,36.48,35.16,19.11,17.19,14.32.HRMS(ESI)(m / z):[M+H] + calc.for C 21 H 19 F2N5O2,412.1584;found 412.1585.

[0080] 8-(3,3-Difluorocyclopentyl)-2-(7-fluoro-1H-indazol-5-yl)amino)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (016) [ka] The synthesis method was the same as in Example 1. Light yellow solid, 39% yield. 1 H NMR(400MHz,DMSO-d6)δ 13.57(s,1H),10.15(s,1H),8.85(d,J=17.4Hz,1H),8.09(s,1H),7.94(s,1H),7.56(d,J=13.1Hz,1H),6.20(d,J=33.6Hz,1H) ,6.19-6.02(m,1H),3.04(dq,J=23.9,12.0Hz,1H),2.40(s,3H),2.38-2.18(m,2H),2.18-1.97(m,3H).HRMS(ESI)(m / z):[M+H]+ calcd for C 20 H 17 F3N6O,415.1496;found 415.1495.

[0081] 2-((1H-pyrazole[3,4-b]pyridin-5-yl)amino)-8-(3,3-difluorocyclopentyl)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (017) [ka] The synthesis method was the same as in Example 1. Light yellow solid, 39% yield. 1 H NMR(600MHz,DMSO-d6)δ 13.61(s,1H),10.18(s,1H),8.87(s,1H),8.70(d,J=2.2Hz,1H),8.51(s,1H),8.07(s,1H),6.24(s,1H),6.08-5. 90(m,1H),2.99(tt,J=44.1,22.0Hz,1H),2.49-2.39(m,2H),2.40(s,3H),2.32-2.17(m,1H),2.15-1.93(m,2H). 13 C NMR(151MHz,DMSO-d6)δ 163.05,159.78,157.70,155.50,154.77,148.95,146.57,145.07,143.34,133.29,130. 17,121.13,117.75,114.45,40.50,39.96,36.48,33.90,17.24.HRMS(ESI)(m / z):[M+H] + calcd for C 19 H 17 F2N7O2,398.1542;found 398.1542. 2-((1H-indazol-5-yl)amino)-8-(3-hydroxycyclopentyl)-5-methylpyridone[2,3-d]pyrimidin-7(8H)-one (018) [ka]

[0082] The synthesis method was the same as in Example 1. Light yellow solid, 39% yield. 1 H NMR(400MHz,DMSO-d6)δ 13.00(s,1H),10.04(s,1H),8.85(s,1H),8.20(s,1H),8.02(s,1H),7.56(d,J=22.2Hz,2H),6.22(s,1H),5.93(s ,1H),5.09(d,J=5.7Hz,1H),4.14(s,1H),2.39(s,3H),2.29-2.18(m,1H),2.11-2.15(m,2H),1.89-1.79(m,3H). 13 C NMR(151MHz,DMSO-d6)δ 163.20,159.57,158.24,157.48,155.85,146.36,137.18,133.65,133.03,123. 31,122.01,117.72,110.45,107.24,71.75,49.07,37.23,34.56,25.56,17.17.

[0083] 2-((1H-indazol-5-yl)amino)-5-methyl-8-(cyclohexane)pyrido[2,3-d]pyrimidin-7(8H)-one (019) [ka] The synthesis method was the same as in Example 1.

[0084] 1 H NMR(400MHz,DMSO-d6)δ 13.03(s,1H),10.04(s,1H),8.82(d,J=22.3Hz,1H),7.98(s,1H),7.53(d,J=8.3Hz,2H),6.19(d,J=22.3Hz,1 H),5.36(d,J=32.6Hz,1H),2.53(s,2H),2.37(s,3H),1.88(d,J=30.7Hz,2H),1.56(s,3H),1.41-1.26(m,3H). 2-((1H-indazol-5-yl)amino)-5-methyl-8-(cycloheptane)pyrido[2,3-d]pyrimidin-7(8H)-one (020) [ka]

[0085] The synthesis method was the same as in Example 1. 2-((1H-indazol-5-yl)amino)-5-methyl-8-(tetrahydrofuran-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one (021) [ka]

[0086] The synthesis method was the same as in Example 1. Light yellow solid, 39% yield. 1 H NMR(400MHz,DMSO-d6)δ 13.00(s,1H),9.99(s,1H),8.83(s,1H),8.13(s,1H),8.01(s,1H),7.64-7.43(m,2H),6.21(d,J =1.0Hz,1H),6.12(s,1H),4.13-4.01(m,1H),4.05-3.82(m,4H),2.49-2.40(m,1H),2.38(s,3H). 13 C NMR(151MHz,DMSO-d6)δ 162.88,159.61,157.41,156.04,146.33,137.30,133.69,132.88,123.34,1 22.34,117.42,111.07,110.44,107.26,68.25,64.34,51.26,28.39,17.20.

[0087] Example 2. Biological activity test In vitro inhibitory effect experiments of the compounds provided by the present invention on RSK4 kinase activity are carried out as follows, using the same method for RSK1-3 as for RSK4 (Kashem, MA et al. J. Biomol. Screen. 12, 70-83). In vitro enzyme activity assay: All enzyme reactions were carried out at 30°C for 40 minutes. The 50 μL reaction mixture contained 40 mM Tris, pH 7.4, 10 mM MgCl2, 0.1 mg / mL BSA, 1 mM DTT, 10 μM ATP, 0.2 μg / mL kinase, and 100 μM lipid substrate. Compounds were diluted in 10% DMSO, and 5 μL of the diluted solution was added to each 50 μL reaction, resulting in a final DMSO concentration of 1% in all reactions. Kinase-Glo reagent was added to the reaction system for detection, which measures kinase activity by quantifying the amount of ATP remaining in solution after the kinase reaction. Nonlinear regression was used to determine the IC 50 Values ​​are calculated and each experiment is repeated two or more times.

[0088] The test results are shown in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6]

[0089] Example 3. Cellular biological activity test Representative compounds with excellent kinase inhibitory activity will be tested for antiproliferative activity against various esophageal squamous cell carcinoma cells TE10, KYSE510, and KYSE150 cells and renal cancer cells ACHN cells. The specific experimental procedure is as follows: After cell digestion, the cells were centrifuged at 1000 rpm for 3 minutes to reduce the density to 3 × 104 Adjust the concentration to cells / mL, inoculate 3000 cells per well, and fill the outermost circle with medium. Place the cells overnight in a 37°C, 5% CO2 incubator to allow cell adhesion. First, dilute the 10 mM stock solution to 100 μM with medium, then dilute three-fold with 1% DMSO-containing medium to create eight concentration gradients. Add 10 μL of compound to each well, and fill the remaining wells with 10 μL of 1% DMSO-containing medium. After culturing the cells for 72 hours, add 10 μL of CCK8 reagent to each well. Incubate in the dark for 2 hours and measure the OD at 450 nM using a microplate reader. Cell viability % = (OD of drug-treated cells - OD of blank) / (OD of control cells - OD of blank) × 100%. After three independent replicates, calculate variance and create statistical graphs to analyze the effect of drugs on cell viability. IC 50 Values ​​are fitted and calculated using GraphPad Prism software.

[0090] Cell antiproliferative activity tests show that pyridonopyrimidine compounds have excellent antiproliferative inhibitory activity against renal cancer cells and esophageal squamous cell carcinoma cells, and are more active than BI-D1870. [Table 2]

[0091] Example 4. Anti-cell invasion activity test By carrying out a Transwell experiment on compound 005, it was found that compound 005 could inhibit the invasion of esophageal squamous cell carcinoma cells TE10 in a dose-dependent manner, and the results are shown in Figure 1.

[0092] Example 5. Effect of compounds on esophageal squamous cell carcinoma cell clonogenesis To further verify the effect of compound 005 on the proliferation ability of TE10 cells, in this example, a cell clonogenesis experiment was used to further evaluate the inhibitory effect of compound 005 on the proliferation of TE10 cells. Compared to the control group, the number of TE10 cell colonies formed after the addition of compound 005 was significantly reduced, and compound 005 was able to significantly inhibit the proliferation of TE10 cells at 2.5 μM. When the concentration of compound 005 reached 5 μM and 10 μM, it almost completely inhibited the proliferation of TE10 cells, which was comparable to the ability of BI-D1870 to inhibit TE10 cell clonogenesis. The results are shown in Figure 2.

[0093] Example 6. Effects of intracellular signaling pathways The RSK kinase family is regulated by the ERK signaling pathway and activated by mitogenic stimulation, and RPS6 is phosphorylated at the Ser235 / 236 site. When RSK4 is activated, it phosphorylates its Ser232 site and simultaneously phosphorylates the RSK family substrate RPS6 and the downstream substrate GSK3β of RSK4. As shown in Figure 3, compound 005 affects the RSK signaling pathway and dose-dependently inhibits the phosphorylation of RSK4, GSK3β, and RPS6.

[0094] Example 7. Study of the pharmacokinetic properties of compound 005 According to the pharmacokinetic experimental data, when compound 005 was administered intravenously at 1 mg / kg, the AUC0-t reached 530.13 ng / mL×h, and when administered orally at 10 mg / kg, the AUC0-t reached 3339.12 ng / mL×h. At the same time, the half-life of 005 was 1.97 hours, with adequate clearance (3.27 L / h / kg) and biodistribution (8.42 L / kg), and the oral bioavailability reached 63%. [Table 3]

[0095] Discussion: After extensive and thorough research, the inventors have designed and synthesized a series of pyrimidopyridone compounds that have not been reported in the literature, and performed molecular level activity tests on the obtained compounds to obtain a group of compounds that can inhibit RSK protein kinase, which will be the basis for the treatment of RSK-mediated cancers.

[0096] All documents mentioned in this application are incorporated by reference in this application as if each document were incorporated by reference individually. Furthermore, after reading the above teachings of the present invention, those skilled in the art will be able to make various changes or modifications to the present invention, and these equivalents will also fall within the scope defined by the appended claims of this application.

Claims

1. A compound of formula I, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, 【Chemical 1】 R １ is hydrogen, optionally substituted C １ -C １０ alkyl groups, optionally substituted C ３ -C ８ Cycloalkyl groups, optionally substituted C ２ -C ６ Alkenyl group, optionally substituted C ３ -C ６ Cycloalkenyl group, optionally substituted C ３ -C ８ Lactone group, optionally substituted C １ -C １０ Amide group, optionally substituted C ５ -C １０ aryl group, optionally substituted C ３ -C ８ Heterocyclic groups, optionally substituted C ５ -C １０ selected from the group consisting of aromatic heterocyclic groups; R ２ is hydrogen, substituted C １ -C １０ Alkyl group, C ３ -C ８ Cycloalkyl groups, optionally substituted C ５ -C １０ aryl group, optionally substituted C ３ -C ８ Heterocyclic groups, optionally substituted C ５ -C １０ selected from the group consisting of an aryl or heteroaryl group and a 5- or 6-membered heterocycle; R ３ is hydrogen, substituted C １ -C １０ Alkyl group, C ３ -C ８ Cycloalkyl group, substituted C １ -C １０ alkylformyl group, optionally substituted C ５ -C １０ Arylformyl group, halogen, cyano group, optionally substituted C ５ -C １０ aryl group, optionally substituted C ３ -C ８ heterocyclic groups; R ４ is hydrogen, optionally substituted C １ -C ６ alkyl groups (e.g., trifluoromethyl groups), optionally substituted C ３ -C ８ Cycloalkyl groups, optionally substituted C １ -C １０ alkylformyl group, optionally substituted C ５ -C １０ arylformyl group, optionally substituted C ５ -C １０ aryl groups, halogens (e.g., fluorine), and optionally substituted C ３ -C ８ A compound represented by formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of heterocyclic groups.

2. R １ is hydrogen, optionally substituted C １ -C １０ alkyl groups, optionally substituted C ３ -C ８ Cycloalkyl groups, optionally substituted C ５ -C １０ aryl group, optionally substituted C ３ -C ８ heterocyclic groups; R ２ is an optionally substituted C ５ -C １０ aryl group, optionally substituted C ３ -C ８ Heterocyclic groups, optionally substituted C ５ -C １０ selected from the group consisting of an aryl or heteroaryl group and a 5- or 6-membered heterocycle; R ３ is hydrogen, optionally substituted C １ -C １０ alkyl groups, optionally substituted C ３ -C ８ cycloalkyl groups, R ４ is hydrogen, optionally substituted C １ -C ６ alkyl groups (e.g., trifluoromethyl groups), optionally substituted C ３ -C ８ cycloalkyl groups, halogen (e.g., fluorine), and optionally substituted C ３ -C ８ heterocyclic groups, 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

3. The compound is a compound of formula II: 【Chemistry 2】 In the formula, R １ is hydrogen, optionally substituted C １ -C １０ alkyl groups, optionally substituted C ３ -C ８ cycloalkyl groups, R ２ is an optionally substituted C ５ -C １０ aryl group, optionally substituted C ３ -C ８ Heterocyclic groups, optionally substituted C ５ -C １０ selected from the group consisting of an aryl or heteroaryl group and a 5- or 6-membered heterocycle; R ４ is hydrogen, optionally substituted C １ -C ６ alkyl groups (e.g., trifluoromethyl groups), optionally substituted C ３ -C ８ cycloalkyl groups selected from the group consisting of 2. The compound according to claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

4. In the formula, R １ is an optionally substituted C ３ -C ８ Cycloalkyl groups, preferably one or more halogen-substituted C ３ -C ８ Cycloalkyl groups, more preferably one or more F-substituted C ３ -C ８ is a cycloalkyl group, R ２ is an optionally substituted C ５ -C １０ an aryl group (preferably a phenyl group), an optionally substituted C ５ -C １０ selected from the group consisting of an aryl group or a heteroaryl 5- or 6-membered heterocycle; Said C ５ -C １０ Aryl group or heteroaryl 5- or 6-membered heterocycle: 【Chemistry 3】 R ４ is hydrogen, optionally substituted C １ -C ６ alkyl groups selected from the group consisting of 4. The compound according to claim 3, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

5. A compound selected from the group consisting of: 【Chemistry 4】 Preferably the following compounds: 【Chemistry 5】

6. 1. A pharmaceutical composition comprising: The pharmaceutical composition comprises the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

7. Use of a compound according to any one of claims 1 to 5 in the preparation of a medicament for treating or preventing a RSK protein kinase mediated disease, or for inhibiting a RSK protein kinase, or for inhibiting one of the RSKs RSK1, RSK2, RSK3 and RSK4.

8. The RSK protein kinase-mediated disease is characterized in that it is cancer. The use according to claim 7.

9. The cancer is selected from the group consisting of esophageal cancer, renal cell carcinoma, pancreatic cancer, colon cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, endometrial cancer, head and neck squamous cell carcinoma, acute myeloid leukemia, and solid tumors, or breast cancer controlled by RSK1 and RSK4, ovarian cancer controlled by RSK3 and RSK4, prostate cancer controlled by RSK1 and RSK2, lung cancer controlled by RSK1, RSK2, and RSK4, head and neck squamous cell carcinoma and acute myeloid leukemia controlled by RSK2, and cancers and solid tumors such as esophageal cancer, renal cancer, endometrial cancer, and colon cancer controlled by RSK4. The use according to claim 8.

10. A method for treating or preventing an RSK-mediated disease using a compound according to any one of claims 1 to 5 or a pharmaceutical composition according to claim 6.

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