Pyrimido five-membered nitrogen heterocyclic compound and its use

Abstract

The present invention relates to pyrimido five-membered nitrogen heterocyclic compounds and their uses. In particular, the present invention relates to pyrimido five-membered nitrogen heterocyclic derivatives represented by general formula (I), methods for preparing the derivatives, pharmaceutical compositions containing the derivatives, and their use as SHP2 inhibitors for preventing and / or treating tumors or cancers. The substituents in general formula (I) have the same definitions as in the specification. #imgabs0#

Description

Technical Field

[0001] The present invention discloses pyrimido five-membered nitrogen heterocyclic compounds, and pharmaceutically acceptable salts, hydrates, prodrugs, stereoisomers, solvates, or isotope-labeled compounds thereof. The present invention also provides methods for preparing such compounds and intermediate compounds thereof, compositions containing such compounds, and the use of such compounds in preparing medicaments for preventing and / or treating diseases or conditions associated with abnormal SHP2 activity. Background Art

[0002] Tyrosine phosphatase SHP2 is composed of two N-terminal Src homology 2 domains (N-SH2 and C-SH2) and a protein tyrosine phosphatase catalytic domain (PTP). In the basal state, N-SH2 binds to PTP to form a ring structure, thereby hindering the binding of PTP to the substrate and inhibiting the enzyme's catalytic activity. When the tyrosine of the upstream receptor protein is phosphorylated, NSH2 binds to it, releasing the PTP catalytic domain and exerting phosphatase activity.

[0003] At the cellular level, SHP2 participates in multiple tumor cell signaling pathways, such as RTK / Ras / MAPK, JAK / STAT, and PB3K / Akt, through its cytoplasmic downstream effects on numerous receptor tyrosine kinases. Through its regulatory effects on these kinases and signaling pathways, SHP2 is closely associated with many important cellular processes, such as cell proliferation, migration, differentiation, death, cytokine regulation, and tumorigenesis.

[0004] SHP2 also participates in programmed death receptor 1 (PD1)-mediated suppression of the immune system. Binding of PD-1 to PD-L1 on T cells recruits a significant amount of SHP2 within the cell. SHP2 dephosphorylates antigen receptor pathway proteins within T cells, thereby inhibiting T cell activation. Therefore, inhibiting SHP2 activity can reverse immune suppression within the tumor microenvironment.

[0005] As an important cell signaling factor, SHP-2 mutations are closely associated with a variety of diseases. Studies have found that SHP-2 mutations are present in neuroblastoma, AML (4%), breast cancer, NSCLC (10%), lung adenocarcinoma (30%), esophageal cancer, head and neck cancer, melanoma, and gastric cancer.

[0006] Currently, several allosteric inhibitors of SHP-2 have entered clinical development, including TNO-155 developed by Novartis, RMC-4630 developed by Revolution Medicine, and JAB-3068 developed by Beijing Jacobs. However, no SHP-2 inhibitor has been developed and marketed for the treatment of Noonan syndrome, Leopard syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, head and neck cancer, lung cancer, and colon cancer. Therefore, the development of a class of druggable SHP-2 inhibitors is urgently needed. Summary of the Invention

[0007] The pyrimido-five-membered nitrogen heterocyclic compounds provided by the present invention are a new class of SHP2 inhibitors that exhibit excellent inhibitory activity against tumor cells and good drugability, with broad prospects for drug development. Furthermore, the preparation method of these compounds is simple, making them suitable for industrial production.

[0008] According to the first aspect of the present invention, the present invention provides a compound as shown in Formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate thereof, or a solvate thereof, or a metabolite thereof, or a prodrug thereof,

[0009]

[0010] in,

[0011] X 1 、X 2 or X 3 Independently selected from CR 5 or N, and at least one of them is N;

[0012] X 4 or X 5 independently selected from C or N;

[0013] R 1 、R 2 、R 3 and R 4 independently selected from H, halogen, hydroxy, amino, cyano, C2-C8 alkenyl, C2-C8 alkynyl, C1-C8 aldehyde, C1-C8 alkyl, C1-C8 heteroalkyl, C3-C8 cycloalkyl, C3-C8 heterocycloalkyl, C1-C8 alkoxy or C1-C3 haloalkoxy, wherein the C1-C8 alkyl, C1-C8 heteroalkyl, C3-C8 cycloalkyl, C3-C8 heterocycloalkyl, C1-C8 alkoxy or C1-C3 haloalkoxy is optionally substituted by one or more R 5 When replaced by multiple R 5 When substituted, R 5 Can be the same or different;

[0014] R 5 is selected from H, halogen, hydroxy, amino, cyano, C1-C3 alkyl, C1-C3 heteroalkyl, C3-C8 cycloalkyl, C3-C8 heterocycloalkyl, C1-C3 alkoxy or C1-C3 haloalkoxy;

[0015] n is selected from 0, 1, 2, 3, 4 and 5;

[0016] m is selected from 0, 1, 2, 3, 4 and 5;

[0017] Ring A is independently selected from C3-C8 cycloalkyl, C6-C 12 Spiroalkyl, C3-C8 heterocycloalkyl, C6-C 10 Aryl or C5-C 12 heteroaryl;

[0018] Ring B is independently selected from C3-C8 cycloalkyl, C3-C8 heterocycloalkyl, C6-C 10 Aryl or C5-C 12 heteroaryl;

[0019] The heteroatoms or heteroatoms contained in the heteroalkyl, heterocycloalkyl and heteroaryl groups are independently selected from -C(=O)N(R 5 )-、-N(R 5 )-, -NH-, -N=, -O-, -S-, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O)2- and -N(R 5 )C(=O)N(R 5 )-; the number of the heteroatoms or heteroatom groups is independently selected from 1, 2 and 3.

[0020] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 1 and X 3 When N, X 2 For C.

[0021] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 2 and X 3 When N, X 1 For C.

[0022] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 1 and X 2 When it is C, X 3 is N.

[0023] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 4 When it is N, ring A is selected from a pyrrole ring, a spiro pyrrole ring, a piperidine ring, a spiro piperidine ring or a dihydropyrazole ring.

[0024] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 4 When N, the ring A is independently selected from

[0025] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 4 When it is C, ring A is selected from a benzene ring, a pyridine ring, a pyrimidine ring, a pyrazine ring or a pyridazine ring.

[0026] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 4 When C, the ring A is independently selected from

[0027] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 5 When is N, ring B is independently a pyrrole ring, a piperidine ring, a piperazine ring or a morpholine ring.

[0028] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 5 When N, ring B is independently

[0029] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 5 When C, ring B is independently a benzene ring, a pyridine ring, a pyrimidine ring, a pyrazine ring or a pyridazine ring.

[0030] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, when X 5 When C, ring B is independently selected from

[0031] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, R 1 、R 2 、R 3 and R 4is selected from H, halogen, hydroxy, amino, cyano, C2-C4 alkenyl, C1-C4 alkyl, C1-C4 heteroalkyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, C1-C4 alkoxy or C1-C3 haloalkoxy, wherein the C1-C4 alkyl, C1-C4 heteroalkyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, C1-C4 alkoxy or C1-C3 haloalkoxy is optionally substituted by one or more R 5 When replaced by multiple R 5 When substituted, R 5 Can be the same or different; where R 5 Selected from H, F, Cl, Br, hydroxy, amino, cyano, methyl, ethyl, cyclopropyl, methoxy, ethoxy, trifluoromethoxy.

[0032] In one preferred embodiment, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) of the present invention, R 1 、R 2 、R 3 and R 4 Selected from H, halogen, hydroxy, amino, cyano, methyl, ethyl, isopropyl, cyclopropyl, vinyl, methoxy, ethoxy, isopropoxy, trifluoromethyl, difluoromethyl, trifluoromethoxy, -OCH2CHF2, -N(CH3)2, -NH(CH3) or -OCH2CF3.

[0033] In some embodiments, the compound represented by formula (I) of the present invention is selected from the compounds represented by the following formula (II), formula (III) or formula (IV):

[0034]

[0035]

[0036] Among them, R 1 、R 2 、R 3 、R 4 、X 4 、X 5 , Ring A and Ring B are as defined in claim 1, m is selected from 0, 1, 2, 3, 4 and 5, and n is selected from 0, 1, 2, 3, 4 and 5.

[0037] In some embodiments, the compound represented by formula (I) of the present invention is selected from the compounds represented by the following formulas (II-1) to (IV-3):

[0038]

[0039]

[0040] Among them, R1 、R 2 、R 3 、R 4 、X 4 、X 5 , Ring A and Ring B are as defined in claim 1, m is selected from 0, 1, 2, 3, 4 and 5, and n is selected from 0, 1, 2, 3, 4 and 5.

[0041] According to a specific embodiment of the present invention, the compound represented by formula I of the present invention is any of the following compounds:

[0042]

[0043] In the present invention, those skilled in the art can select the groups and substituents thereof in the compound represented by Formula I to provide a stable compound represented by Formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate thereof, or a solvate thereof, or a metabolite thereof, or a prodrug thereof, including but not limited to I-1 to I-6 described in the embodiments of the present invention.

[0044] The reaction solvents used in each reaction step described in the present invention are not particularly limited. Any solvent that can dissolve the starting materials to a certain extent and does not inhibit the reaction is included in the present invention. In addition, many similar modifications, equivalent substitutions, or solvents, solvent combinations, and different ratios of solvent combinations equivalent to those described in the present invention are considered to be within the scope of the present invention.

[0045] According to the second aspect of the present invention, the present invention provides a pharmaceutical composition comprising an effective dose of a compound represented by Formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate thereof, or a solvate thereof, or a metabolite thereof, or a prodrug thereof, and at least one pharmaceutical excipient.

[0046] The pharmaceutical excipients may be those widely used in the field of pharmaceutical production. Excipients are primarily used to provide a safe, stable, and functional pharmaceutical composition. They may also provide methods to dissolve the active ingredient at a desired rate after administration to a subject, or to promote effective absorption of the active ingredient after administration of the composition to a subject. The pharmaceutical excipients may be inert fillers or provide a function, such as stabilizing the overall pH of the composition or preventing degradation of the active ingredient. The pharmaceutical excipients may include one or more of the following: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

[0047] The pharmaceutical compositions of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0048] The pharmaceutical compositions of the present invention can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ophthalmic, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical compositions of the present invention can also be controlled-release or delayed-release dosage forms (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions. Examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols such as nasal sprays or inhalers; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

[0049] Oral administration of the compounds of the present invention is preferred. Intravenous administration of the compounds of the present invention is also preferred. Depending on the circumstances, other routes of administration may be applicable or even preferred. For example, transdermal administration may be highly desirable for patients who are forgetful or irritable with oral medications. In special circumstances, the compounds of the present invention may also be administered transdermally, intramuscularly, intranasally, or intrarectally. The route of administration may vary in any manner, subject to the physical properties of the drug, the convenience of the patient and caregiver, and other relevant circumstances.

[0050] According to a third aspect of the present invention, the present invention provides use of a compound represented by Formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a pharmaceutical composition thereof, for preparing a medicament for treating and / or preventing diseases caused by abnormal SHP2 mutations. The compounds provided by the present invention can be used to treat and / or prevent one or more diseases caused by abnormal SHP2 mutations and have promising clinical and medical applications.

[0051] According to a fourth aspect of the present invention, the present invention provides a compound as represented by Formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, metabolite, prodrug, or pharmaceutical composition thereof, for use in preparing a SHP2 inhibitor. The compound provided by the present invention has excellent SHP2 enzymatic activity and cell proliferation inhibition activity, and can be effectively used as a SHP2 inhibitor, and as a therapeutic drug for SHP2 inhibitors.

[0052] According to a fifth aspect of the present invention, the present invention provides a compound as shown in Formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate thereof, or a solvate thereof, or a metabolite thereof, or a prodrug thereof, or a pharmaceutical composition thereof for use in the preparation of a drug for treating and / or preventing cancer. The compound provided by the present invention can be used to prepare a drug for treating and / or preventing cancer, wherein the cancer includes but is not limited to Noonan syndrome, Leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, breast cancer, esophageal cancer, lung cancer, colon cancer, gastric cancer, anaplastic large cell lymphoma, glioblastoma, non-small cell lung cancer or head and neck tumors.

[0053] The present invention has the following advantages:

[0054] 1. The pyrimido-5-membered nitrogen heterocyclic compounds disclosed in this invention are a novel class of allosteric inhibitors that can inhibit SHP2 activity by binding to the non-catalytic region and "locking" it into its weak basal state. These pyrimido-5-membered nitrogen heterocyclic compounds overcome the common shortcomings of PTP catalytic region inhibitors, such as poor selectivity and drugability, and exhibit excellent biological activity and drugability, suggesting promising drug development prospects.

[0055] 2. In the evaluation systems such as SHP2 enzyme activity inhibition test, phosphorylated protein kinase (p-ERK) cell test, NCI-H358 cell and MV-4-11 cell proliferation inhibition test under the same conditions, the present invention showed superior activity and PK properties compared with the compounds RMC4550 and TNO155 with disclosed structures.

[0056] Terms and Definitions

[0057] Unless stated otherwise, the following terms used in the specification and claims have the following meanings.

[0058] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, for example, including straight and branched chain groups of 1 to 20 carbon atoms, for example, straight and branched chain groups of 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. "Alkyl" herein can be a monovalent, divalent, or trivalent group. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, and various branched chain isomers thereof. Non-limiting examples also include methylene, methine, ethylene, ethylene, propylene, propylene, butylene, butylene and various branched chain isomers thereof. Alkyl can be optionally substituted or unsubstituted.

[0059] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, for example, comprising 3 to 12 ring atoms, for example, 3 to 12, 3 to 10 or 3 to 6 ring atoms, or a 3, 4, 5, or 6-membered ring. The non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Cyclic radicals can be optionally substituted or unsubstituted.

[0060] The term "spirocycloalkyl" refers to a cycloalkyl group in which two rings share one ring atom. The shared ring atom is also called a spiro atom, most commonly a quaternary carbon (spiro carbon).

[0061] The term "heteroalkyl" refers to a stable straight or branched chain consisting of a specified number of carbon atoms and one or more heteroatoms selected from the group consisting of O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2-S(O)-CH3, -CH2-CH2-S(O)2-CH3, or -CH2-CH=N-OCH3. Up to two heteroatoms may be consecutive, for example, -CH2-NH-O-CH3. In certain embodiments, heteroalkyl is optionally substituted as described elsewhere herein.

[0062] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, for example, comprising 3 to 20 ring atoms, for example 3 to 16, 3 to 12, 3 to 10 or 3 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m In some embodiments, the heteroatoms of heterocycloalkyl include pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. The limiting examples of heterocycloalkyl include pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. The limiting examples of heterocycloalkyl include spirocyclic, condensed ring or bridged ring.

[0063] The term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one double bond. Non-limiting examples of alkenyl include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1,3-butadienyl, and the like.

[0064] The term "alkynyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one triple bond. Non-limiting examples of alkynyl groups include, but are not limited to, ethynyl (-C≡CH), 1-propynyl (-C≡C-CH3), 2-propynyl (-CH2-C≡CH), 1,3-butadiynyl (-C≡C≡CH), and the like.

[0065] The term "alkoxy" refers to an -O-alkyl group.

[0066] The term "haloalkoxy" refers to an alkoxy group substituted by one or more halogen atoms which may be the same or different, such as trifluoromethoxy.

[0067] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. For example, an aryl group can have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and 1,2,3,4-tetrahydronaphthalene.

[0068] The term "heteroaryl" refers to the residue remaining after removing one hydrogen atom from a "heteroaromatic ring" molecule. A heteroaryl group may be unsubstituted or substituted, and the substituents include, but are not limited to, alkyl, alkyloxy, aryl, aralkyl, amino, halogen, hydroxy, cyano, nitro, carbonyl, and heteroalicyclic groups. Non-limiting examples of unsubstituted heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, quinolyl, isoquinolyl, tetrazolyl, and triazinyl.

[0069] The term "halogen" refers to fluorine, chlorine, bromine, and iodine, with fluorine, chlorine, and bromine being preferred in some embodiments.

[0070] The term "amino" refers to -NH2 group, -NH(alkyl) and -N(alkyl)2. Specific examples of amino include, but are not limited to, -NH2, -NHCH3, -NHCH(CH3)2, -N(CH3)2, -NHC2H5, -N(CH3)C2H5, etc.

[0071] The term "hydroxy" refers to an -OH group.

[0072] The term "cyano" refers to a -CN group.

[0073] The term "optional" or "optionally" means that the event or situation described subsequently may or may not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may or may not be present, i.e., including the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group.

[0074] "Substituted" means that one or more hydrogen atoms in the group are replaced by a substituent. For example, up to 5, more preferably 1-3 hydrogen atoms in the group are independently replaced by a corresponding number of substituents.

[0075] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids such as arginine, and organic acids such as glucuronic acid (see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts. Preferably, salt is contacted with alkali or acid in a conventional manner, then the parent compound is separated, thus regenerating the neutral form of the compound. The difference between the parent form of the compound and the form of its various salts is some physical properties, such as different solubility in polar solvents. According to embodiments of the present invention, it is preferred that the compound shown in the formula I of the present invention is hydrochloride, hydrobromide, phosphate or sulfate in a pharmaceutically acceptable salt, most preferably hydrochloride.

[0076] The term "solvate" refers to a physical association of a compound of the invention with one or more solvent molecules. This physical association includes various degrees of ionic and covalent bonding, including hydrogen bonding. "Solvate" includes both solution-phase and isolatable solvates. Non-limiting examples of solvates include ethanolates, methanolates, and the like. "Hydrate" is a solvate in which the solvent molecule is HO.

[0077] A "pharmaceutical composition" refers to a mixture containing one or more compounds, or pharmaceutically acceptable salts, stereoisomers, or tautomers thereof, and other chemical components, as well as other components such as pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0078] Any formula or structure given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have the structures shown by the formulas given herein, except that one or more atoms are replaced by atoms having a selected atomic mass or mass number. Examples of isotopes that can be introduced into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine, such as, but not limited to, 2 H (deuterium, D), 3 H (tritium), 11 C. 13 C. 14 C. 15 N. 18 F. 35 S. 36 Cl and 125 I. Various isotopically labeled compounds disclosed herein, for example, 3 H. 13 C and 14 C and other radioactive isotopes. Such isotope-labeled compounds can be used for metabolic studies, reaction kinetics studies, detection or imaging techniques, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT) including drug or substrate tissue distribution analysis, or for radiotherapy of patients. Isotope-labeled compounds of the present disclosure and prodrugs thereof can generally be prepared by carrying out the processes disclosed in the schemes or in the examples and preparations described below, by substituting readily available isotope-labeled reagents for non-isotope-labeled reagents.

[0079] The compound of the present invention can be in the form of a prodrug compound." prodrug compound" means by reacting with enzyme or gastric acid etc. under physiological conditions in an organism, for example, by oxidation, reduction or hydrolysis etc., and is converted into a derivative of the compound according to the present invention, and each of the reactions is carried out enzymatically. The example of a prodrug is the following compound, wherein the amino in the compound of the present invention is acylated, alkylated or phosphorylated to form, for example, eicosanoylamino, alanylamino, pivaloyloxymethylamino, or wherein hydroxyl is acylated, alkylated, phosphorylated or converted into boric acid ester, for example, acetyloxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaryloxy, alanyloxy, or wherein carboxyl is esterified or amidated. These compounds can be produced from the compound of the present invention according to known methods. Other examples of a prodrug are the following compound, wherein, for example, the carboxylate in the compound of the present invention is converted into alkyl-, aryl-, choline-, amino, acyloxymethyl ester, linolenoyl ester.

[0080] In the case of compounds of the present invention or their prodrugs, where tautomerism, such as, for example, keto-enol tautomerism, occurs, the individual forms, such as, for example, keto and enol forms, as well as mixtures thereof in any proportion, are each within the scope of the present invention. The same applies to stereoisomers, such as, for example, enantiomers, cis / trans isomers, and conformers.

[0081] If desired, isomers can be separated by methods known in the art, such as by liquid chromatography. The same applies to enantiomers by using, for example, a chiral stationary phase. In addition, enantiomers can be separated by converting them into diastereomers, i.e., coupling with an enantiomerically pure auxiliary compound, subsequently separating the resulting diastereomers and cleaving the auxiliary residue. Alternatively, any enantiomer of the compounds of the present invention can be obtained from stereoselective synthesis using optically pure starting materials.

[0082] The term "treatment" means administering the compound or formulation of the present invention to prevent, improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0083] (i) preventing a disease or disease state from occurring in a mammal, particularly where such mammal is susceptible to the disease state but has not yet been diagnosed as having the disease state;

[0084] (ii) inhibiting the disease or disease state, i.e., curbing its development;

[0085] (iii) ameliorating the disease or condition, i.e., causing regression of the disease or condition.

[0086] The term "effective dose" means an amount of a compound of the present invention that (i) treats or prevents a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of a compound of the present invention that constitutes an "effective dose" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art based on their own knowledge and this disclosure.

[0087] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention.

[0088]

[0089] The specific reaction conditions are described as follows:

[0090] Compound A and Compound B react under basic conditions, such as potassium carbonate, DIEA, cesium carbonate, and sodium tert-butoxide, in a solvent such as DMF, DMSO, or NMP, to produce Compound C. Compound C and Boc anhydride react in dichloromethane with DMAP as a catalyst to produce Compound D. Compound D and Compound E react using Pd2(dba)3 as a catalyst, Xantphos as a ligand, and an organic or inorganic base, such as DIEA or cesium carbonate, to produce Compound F. Compound F is deprotected with TFA or ethyl hydrochloride to produce the compound of Formula I in free or hydrochloride form. DETAILED DESCRIPTION

[0091] The compound of formula I of the present invention, or its pharmaceutically acceptable salt, or its stereoisomer, or its tautomer, or its hydrate, or its solvate, or its metabolite, or its prodrug can be prepared by the exemplary methods described in the following examples and the relevant public literature operations used by those skilled in the art, but these examples do not limit the scope of the present invention.

[0092] The structures of the compounds of the present invention were determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker AVANCE-400 or Varian Oxford-300 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDC13), or deuterated methanol (CD3OD) as the solvent, tetramethylsilane (TMS) as the internal standard, and chemical shifts measured in 10 -6 The units are given in ppm.

[0093] MS was measured using an Agilent SQD (ESI) mass spectrometer (manufacturer: Agilent, model: 6110) or a Shimadzu SQD (ESI) mass spectrometer (manufacturer: Shimadzu, model: 2020).

[0094] HPLC analysis was performed using an Agilent 1200DAD high pressure liquid chromatograph (Sunfirc C18, 150×4.6 mm, 5 μm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18, 150×4.6 mm, 5 μm column).

[0095] The thin layer chromatography silica gel plate used was Qingdao Ocean GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.15mm-0.2mm, and the specification used for thin layer chromatography separation and purification products was 0.4mm-0.5mm silica gel plate.

[0096] Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as the carrier.

[0097] The known starting materials of the present invention can be synthesized by methods known in the art, or purchased from companies such as Accela ChemBio Inc. and Beijing Coupling Chemicals.

[0098] Unless otherwise specified in the examples, the reactions were carried out under an argon or nitrogen atmosphere. The hydrogenation reaction was usually carried out by evacuating the air and then filling with hydrogen, and the operation was repeated three times.

[0099] Unless otherwise specified in the examples, the reaction temperature is room temperature, which ranges from 5°C to 35°C.

[0100] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent systems used in the reactions were A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compounds.

[0101] The eluent system for column chromatography and the developing solvent system for thin-layer chromatography used for purifying compounds include A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvents is adjusted according to the polarity of the compound, and a small amount of triethylamine and acidic or alkaline reagents can also be added for adjustment.

[0102] The present invention is described in detail below by way of examples, but this does not imply any adverse limitations on the present invention. The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. The following synthetic schemes describe the steps for preparing the compounds disclosed herein. Unless otherwise indicated, each substituent has the definition as described herein.

[0103] Synthesis of intermediates:

[0104] Synthesis of intermediate Q1

[0105]

[0106] Step 1: Synthesis of Q1-2

[0107] Compound Q1-1 (100 g, 495 mmol) was added to DME (1000 ml). The temperature was cooled to -10°C, and isobutyl chloroformate (67.6 g, 495 mmol) and 4-methylmorpholine (50 g, 495 mmol) were added. The mixture was allowed to react at room temperature for 5 hours. After completion of the reaction by TLC, the mixture was filtered and the solid was washed with DME (250 ml). The filtrate was then added to a 2 L three-necked flask and treated with sodium borohydride (37.6 g, 990 mmol). The mixture was stirred at room temperature for 30 minutes, and methanol (250 ml) was slowly added dropwise. The reaction was continued at room temperature for 3 hours. After completion of the reaction by TLC, the reaction solution was dried by rotary evaporation, and water (1500 ml) was added. The aqueous phase was extracted with DCM (300 ml x 3). The organic phases were combined, washed sequentially with water and saturated sodium chloride, and then dried and rotary evaporation to obtain compound Q1-2 (86.5 g, white solid) in a 93% yield. This was used directly in the next step.

[0108] Step 2: Synthesis of Q1-3

[0109] Compound Q1-2 (25 g, 133 mmol) was added to DCM (250 ml), followed by TEA (26.9 g, 266 mmol). MsCl (18.3 g, 156 mmol) was added dropwise in an ice bath, and the mixture was allowed to react at room temperature for 1 h. After TLC indicated completion of the reaction, the reaction solution was diluted with DCM, washed sequentially with water and then with saturated sodium chloride. The organic phase was dried, concentrated, and the resulting residue was purified by silica gel chromatography (eluent: petroleum ether:ethyl acetate = 3:1 to 1:1) to afford compound Q1-3 (33.6 g, colorless, transparent liquid) in a 95% yield.

[0110] MS m / z(ESI):266[M+1].

[0111] Step 3: Synthesis of Q1-4

[0112] N-Boc-4-cyanopiperidine (28 g, 130 mmol) was added to THF (300 ml), cooled to -78 ° C, and 2.0 M LDA (75 ml, 150 mmol) was slowly added dropwise. After the addition was complete, the reaction was continued at -78 ° C for 1.5 h. Then, a THF solution (150 ml) of the compound Q1-3 (26.6, 100 mmol) obtained in the first step was added dropwise. After the addition was complete, the reaction was continued at -78 ° C for 3 h. TLC After the reaction was completed, saturated ammonium chloride solution (50 ml) was added dropwise to quench the reaction, and saturated sodium chloride aqueous solution (500 ml) was added to separate the organic phase, and the aqueous phase was extracted with ethyl acetate (150 ml * 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was filtered through a silica gel chromatography column (eluent petroleum ether: ethyl acetate = 10:1 to 1:1) to obtain compound Q1-4 (25.7 g, white solid) with a yield of 67.6%.

[0113] MS m / z(ESI):380[M+1].

[0114] Step 4: Synthesis of Q1-5

[0115] Compound Q1-4 (25 g, 65.8 mmol) obtained in the previous step was added to a mixed solvent of DMA (200 ml) and water (20 ml). Triethylamine (33.2 g, 329 mmol) and the catalyst dichlorodi-tert-butyl-(4-dimethylaminophenyl)phosphine palladium(II) (4.6 g, 6.6 mmol, CAS: 887919-35-9) were then added. The mixture was reacted at 130°C under nitrogen for 4 hours. After TLC indicated completion of the reaction, the reaction solution was cooled and diluted with water (800 ml). The aqueous phase was extracted with ethyl acetate (200 mL × 3). The organic phases were combined and washed with saturated brine (200 mL × 2). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The resulting residue was purified by silica gel chromatography (eluent: petroleum ether:ethyl acetate = 3:1 to 1:1) to afford compound Q1-5 (14.6 g, white solid) in a 73% yield.

[0116] MS m / z(ESI):303[M+1].

[0117] Step 5: Synthesis of Q1-6

[0118] Compound Q1-5 (10 g, 33 mol) obtained in the previous step was added to tetraethyl titanate (100 ml), and then (R)-(+)-tert-butylsulfenamide (4.8 g, 40 mml) was added. The temperature was raised to 90°C and the reaction was carried out for 3 hours. After TLC showed that the reaction was complete, the mixture was cooled to room temperature and the reaction solution was slowly added to ice water (500 ml). The obtained aqueous phase was extracted with dichloromethane (150 mL×3), and the organic phases were combined and washed with saturated brine (100 mL×2). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was filtered through a silica gel chromatography column (eluent petroleum ether: ethyl acetate = 3:1 to 1:1) to obtain compound Q1-6 (12 g, yellow solid) with a yield of 89.6%.

[0119] MS m / z(ESI):406[M+1].

[0120] Step 6: Synthesis of Q1-7

[0121] Compound Q1-6 (10 g, 24.6 mmol) obtained in the previous step was added to tetrahydrofuran (100 ml). After cooling to -78°C, DIBAL-H (30 ml, 30 mmol, 1 M toluene solution) was slowly added dropwise, and the reaction was continued at -78°C for 0.5 h. After TLC showed that the reaction was complete, saturated Rochelle salt solution (300 ml) was added at -50°C to quench the reaction. The mixture was stirred at room temperature for 30 min, and the obtained aqueous phase was extracted with ethyl acetate (150 mL×3). The organic phases were combined and washed with saturated brine (100 mL×2). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was filtered through a silica gel chromatography column (eluent petroleum ether: ethyl acetate = 3:1 to 1:1) to obtain compound Q1-7 (8.6 g, white solid) with a yield of 85.3%.

[0122] MS m / z(ESI):408[M+1].

[0123] Step 7: Synthesis of intermediate Q1

[0124] Compound Q1-7 (5 g, 12.2 mmol) obtained in the previous step was added to ethyl acetate (50 ml), and then 4 M HCl / ethyl acetate solution (25 ml) was added. The reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was complete, the mixture was filtered, and the solid was washed with ethyl acetate and dried to obtain the hydrochloride salt of compound Q1 (3.2 g, white solid) with a yield of 94%.

[0125] MS m / z(ESI):204[M+1].

[0126] Synthesis of intermediate Q2

[0127]

[0128] Step 1: Synthesis of Q2-2

[0129] 5-Bromo-2,4-dichloropyrimidine (5.0 g, 21.94 mmol) was added to an ethanol solution (100 mL), followed by triethylamine (3.7 mL, 26.33 mmol) and 2,2-dimethoxyethylamine (2.53 mL, 24.14 mmol). The reaction mixture was stirred at room temperature for 24 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure. 200 mL of water was added, and the resulting aqueous phase was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and washed with saturated brine (70 mL × 2). The organic phases were then dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The resulting residue was purified by silica gel chromatography (eluent: petroleum ether:ethyl acetate = 3:1 to 1:1) to obtain compound Q2-2 (5.4 g, white solid) in 83% yield.

[0130] MS m / z(ESI):296[M+1].

[0131] Step 2: Synthesis of Q2-3

[0132] Compound Q2-2 (5.4 g, 18.2 mmol) obtained in the previous step was dissolved in concentrated sulfuric acid (50 mL) and then heated to 75°C. After 2 h of reaction, TLC indicated the reaction was complete. After cooling to room temperature, the reaction solution was slowly poured into ice water and then slowly basified to approximately pH 6 with 5 M aqueous sodium hydroxide solution. A large amount of solid precipitated, which was filtered, the filter cake was washed with water, and the solid was dried to obtain compound Q2-3 (3.2 g, gray solid) in an 82.5% yield.

[0133] MS m / z(ESI):214[M+1].

[0134] Step 3: Synthesis of intermediate Q2

[0135] Phosphorus oxychloride (50 mL) and then DIEA (5.8 g, 45 mmol) were added to compound Q2-3 (3.2 g, 15 mmol), and the mixture was heated to 115°C. After 3 h of reaction, TLC indicated the reaction was complete. The mixture was concentrated in vacuo, and the resulting residue was diluted with EtOAc (100 mL). The pH was then slowly adjusted to 7 with saturated sodium bicarbonate solution. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated brine (70 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The resulting residue was then purified by flash silica gel chromatography (eluent: petroleum ether:ethyl acetate = 3:1 to 1:1) to afford compound Q2 (2.7 g, white solid) in a 78% yield.

[0136] MS m / z(ESI):232[M+1].

[0137] Synthesis of intermediate Q3

[0138]

[0139] Step 1: Synthesize Q3-2

[0140] 5-Iodo-2,4-dichloropyrimidine (16.3 g, 59.2 mmol) and hydrazine hydrate (8.8 mL, 181 mmol) were added to anhydrous ethanol (300 mL) and heated under reflux for 12 hours. After TLC indicated completion of the reaction, the mixture was cooled to 0°C, whereupon a large amount of solid precipitated. The solid was filtered, and the filter cake was washed with icy ethanol (100 mL) and dried to afford Q3-2 (15.1 g, yellow solid) in a 94.3% yield. This solid was used in the next step without further purification.

[0141] MS m / z(ESI):271[M+1].

[0142] Step 2: Synthesis of intermediate Q3

[0143] Compound Q3-2 (15 g, 55.3 mmol) obtained in the previous step was added to trimethyl orthoformate (100 ml), heated under reflux for 12 hours, and TLC showed that the reaction was completed. The mixture was cooled to 0°C, and a large amount of solid precipitated. The filter cake was filtered and washed with ice 50% ethanol (50 mL) and dried to obtain compound Q3 (11.3 g, yellow solid) with a yield of 88%. It can be used in the next step without further purification.

[0144] MS m / z(ESI):281[M+1].

[0145] Synthesis of intermediate Q4

[0146]

[0147] Step 1: Synthesis of compound Q4-2

[0148] Compound Q4-1 (40 g, 192 mmol) was added to N,N-dimethylformamide dimethyl acetal (200 ml), and the temperature was raised to 120°C for 3 hours. After TLC showed completion of the reaction, most of the solvent was removed under reduced pressure, and then water (200 ml) was added, resulting in the precipitation of a large amount of solid. The solid was collected by filtration, and the filter cake was washed with dichloromethane and dried to obtain compound Q4-2 (42 g, white solid) in an 82% yield. This was used in the next step without further purification.

[0149] MS m / z(ESI):263[M+1].

[0150] Step 2: Synthesis of compound Q4-3

[0151] Compound Q4-2 (42 g, 160 mmol) obtained in the previous step was added to methanol (250 mL), followed by the addition of hydroxylamine hydrochloride (13.4 g, 192 mmol). The reaction solution was stirred at room temperature for 3 hours. After TLC showed that the reaction was complete, the reaction solution was cooled to 0°C, and a large amount of solid precipitated. The solid was filtered, and the filter cake was washed with 100 mL of ice-cold methanol. The solid was dried to obtain compound Q4-3 (33.6 g, white solid) with a yield of 84%. It was used in the next step without further purification.

[0152] MS m / z(ESI):251[M+1].

[0153] Step 3: Synthesis of compound Q4-4

[0154] Compound Q4-3 (7.5 g, 29.6 mmol) obtained in the previous step was added to polyphosphoric acid (50 ml), the reaction temperature was raised to 120 ° C, and the reaction was stirred for 5 hours. After TLC showed that the reaction was completed, 150 mL of ice water was added to the reaction system, and the pH was adjusted to about 8 with 2N sodium hydroxide aqueous solution. The obtained aqueous phase was then extracted with n-butanol (150 mL*3), and the organic phases were combined. The combined organic phases were washed with saturated brine (70 mL×2), and then the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain compound Q4-4 (3.5 g, light yellow solid) with a yield of 50%.

[0155] MS m / z(ESI):215[M+1].

[0156] Step 4: Synthesis of intermediate Q4

[0157] Phosphorus oxychloride (50 mL) and then DIEA (5.8 g, 45 mmol) were added to compound Q4-4 (3.2 g, 15 mmol), and the mixture was heated to 115°C. After 3 h of reaction, TLC indicated the reaction was complete. The mixture was concentrated in vacuo, and the resulting residue was diluted with EtOAc (100 mL). The pH was then slowly adjusted to 7 with saturated sodium bicarbonate solution. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated brine (70 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The resulting residue was then purified by flash silica gel chromatography (eluent: petroleum ether:ethyl acetate = 3:1 to 1:1) to afford compound Q4 (2.5 g, white solid) in a 71.5% yield.

[0158] MS m / z(ESI):233[M+1].

[0159] Synthesis of intermediate Q5

[0160]

[0161] Step 1: Synthesis of compound Q5-2

[0162] Compound Q5-1 (10 g, 88.4 mmol) was added to a THF solution (100 ml), followed by butyl 2-hydroxymethylpyrrolidine-1-carboxylate (21.4 g, 106.1 mmol, CAS: 170491-63-1) and triphenylphosphine (34.8 g, 132.6 mmol). The reaction solution was cooled to below 5°C in an ice-water bath, and then DEAD (23.1 g, 132.6 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 16 hours. TLC indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by flash silica gel chromatography (eluent: petroleum ether:ethyl acetate = 10:1) to afford compound Q5-2 (25 g, light yellow oil) in a 95.4% yield.

[0163] MS m / z(ESI):297[M+1].

[0164] Step 2: Synthesis of compound Q5-3

[0165] Compound Q5-2 (5 g, 16.8 mmol) obtained in the previous step was added to ethyl acetate (50 ml), and then 4 M HCl / ethyl acetate solution (25 ml) was added. The reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, the mixture was filtered, and the solid was washed with ethyl acetate and dried to obtain the hydrochloride salt of compound Q5-3 (4.4 g, white solid) with a yield of 96.8%.

[0166] MS m / z(ESI):197[M+1].

[0167] Step 3: Synthesis of compound Q5-4

[0168] The hydrochloride salt of compound Q5-3 (2.20 g, 8.17 mmol) obtained in the previous step was added to ethanol (50 mL), followed by potassium carbonate (5.64 g, 40.87). After stirring at room temperature for half an hour, the mixture was heated to 65°C and stirred for 12 hours. TLC indicated the reaction was complete. The mixture was then filtered and the filtrate was concentrated under reduced pressure. The resulting residue was then purified by flash silica gel chromatography (eluent: petroleum ether:ethyl acetate = 10:1) to afford compound Q5-4 (1.22 g, colorless oil) in an 84.7% yield.

[0169] MS m / z(ESI):177[M+1].

[0170] Step 4: Synthesis of compound Q5-5

[0171] Compound Q5-4 (1.2 g, 6.8 mmol) obtained in the previous step was dissolved in THF (20 mL) and then cooled to -78°C. Under a nitrogen atmosphere, n-butyllithium solution (5.4 mL, 13.6 mmol, 2.5 M in hexane) was slowly added dropwise. After the addition was complete, the mixture was naturally warmed to 0°C and stirred for 1.5 hours. The reaction mixture was then cooled to -78°C again, and a solution of elemental iodine (2 g, 8.16 mmol) in THF (10 mL) was added dropwise to the mixture. After the addition was complete, the reaction mixture was naturally warmed to room temperature and stirred at room temperature for 2 hours. After TLC showed that the reaction was completed, saturated NH4Cl aqueous solution (100 ml) was added to the reaction solution, and the obtained aqueous phase was extracted with ethyl acetate (100 mL×3). The organic phases were combined, and the organic phase was washed with saturated brine (70 mL×2). The organic phase was then dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was passed through a silica gel chromatography column (eluent petroleum ether: ethyl acetate = 3:1 to 1:1) to obtain compound Q5-5 (1.5 g, yellow solid) with a yield of 72.8%.

[0172] MS m / z(ESI):303[M+1].

[0173] Step 5: Synthesis of compound Q5-6

[0174] Compound Q5-5 (303 mg, 1 mmol) obtained in the previous step was dissolved in dioxane (5 ml), followed by the addition of methyl 3-mercaptopropionate (180 mg, 1.5 mmol), Pd(dba) (22.73 mg, 0.025 mmol, 0.05 equiv), Xantphos (14.36 mg, 0.025 mmol), and DIEA (387 mg, 3 mmol). The mixture was heated to 90°C and stirred for 1 hour under nitrogen. The reaction mixture was then concentrated under reduced pressure. The resulting residue was dried and purified by silica gel chromatography (eluent: petroleum ether:ethyl acetate = 3:1 to 1:1) to afford compound Q5-6 (239 mg, pale yellow solid) in an 81% yield.

[0175] MS m / z(ESI):295[M+1].

[0176] Step 6: Synthesis of intermediate Q5

[0177] Compound Q5-6 (250 mg, 0.85 mmol) obtained in the previous step was dissolved in THF (2 ml) and then cooled to 0° C. Sodium tert-butoxide (96 mg, 1 mmol) was added, and after stirring at 0° C. for 0.5 hour, TLC showed that the reaction was complete. The reaction mixture was diluted with PE to precipitate a large amount of solid. The precipitated solid was collected by filtration and washed with ethyl acetate to give the intermediate (164 mg, light yellow solid) with a yield of 84%.

[0178] MS m / z(ESI):209[M+1].

[0179] Referring to the synthesis of intermediate Q5, commercially available raw materials were used instead of 2-hydroxymethylpyrrolidine-1-carboxylic acid butyl ester to synthesize the following intermediates:

[0180]

[0181] Example 1: Preparation of the compound represented by formula I-1

[0182]

[0183] Synthesis route:

[0184]

[0185] Step 1: Synthesis of compound 1A

[0186] Compound Q2 (220 mg, 0.95 mmol), Q1 (267 mg, 0.95 mmol), and N,N-diisopropylethylamine (0.47 mL, 2.84 mmol) were dissolved in 5 mL of dimethyl sulfoxide and reacted at 90°C for 1.5 hours. After the reaction, 20 mL of ethyl acetate and 40 mL of water were added, and the mixture was extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated sodium hydroxide solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure, and the residue was passed through a silica gel chromatography column (eluent: dichloromethane:methanol = 50:1 to 10:1) to obtain compound 1A (244 mg, light yellow solid) in a 64.3% yield.

[0187] MS m / z(ESI):399[M+1].

[0188] Step 2: Synthesis of Compound 1B

[0189] Compound 1A (240 mg, 0.6 mmol) obtained in the previous step was added to dichloromethane (5 ml), followed by the addition of DIEA (154 mg, 1.2 mmol) and (Boc)2 (262 mg, 1.2 mmol). The mixture was stirred at room temperature for 12 hours. TLC showed that the reaction was complete. The reaction solution was diluted with 10 ml of dichloromethane, and the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure, and the residue was passed through a silica gel chromatography column (eluent: dichloromethane: methanol = 50:1 to 10:1) to give compound 1B (266 mg, light yellow solid) in an 89% yield.

[0190] MS m / z(ESI):499[M+1].

[0191] Step 3: Synthesis of Compound 1C

[0192] Compound 1B (250 mg, 0.5 mmol) obtained in the previous step was dissolved in dioxane (5 ml), and intermediate Q5 (230 mg, 1 mmol), Pd2(dba)3 (22.73 mg, 0.025 mmol), Xantphos (14.36 mg, 0.025 mmol), and DIEA (190 mg, 1.5 mmol) were added. After stirring at 90°C under nitrogen for 1 hour, the reaction mixture was concentrated under reduced pressure. The resulting residue was spin-dried and purified by silica gel chromatography (eluent: dichloromethane:methanol = 50:1 to 10:1) to obtain compound 1C (241 mg, light yellow solid) in a 77% yield.

[0193] MS m / z(ESI):627[M+1].

[0194] Step 4: Synthesis of Compound I-1

[0195] Compound 1C (200 mg, 0.32 mmol) obtained in the previous step was added to ethyl acetate (5 ml), and then 4 M HCl / ethyl acetate solution (2 ml) was added. The reaction was allowed to react at room temperature for 1 h. After TLC showed the reaction was complete, the mixture was filtered, and the solid was washed with ethyl acetate and dried to obtain the hydrochloride salt of compound I-1 (157 mg, yellow solid) with a yield of 87%.

[0196] MS m / z(ESI):527[M+1].

[0197] HNMR: (400MHz, DMSO)8.55-8.54(m,1H),8.48(brs,2H),8.14(s,1H),8.03-7.94 (m,1H),7.92(s,1H),7.84(s,1H),7.37-7.34(m,2H),5.92-5.90(m,1H),4.75-4 .72(m,1H),4.54(brs,1H),4.04-3.93(m,2H),3.77-3.75(m,2H),3.66-3.57(m, 5H).3.25-3.22(m,2H),3.16-3.11(m,1H),2.17-1.93(m,5H),1.74-1.52(m,3H).

[0198] Example 2: Preparation of the compound represented by formula I-2

[0199]

[0200] The synthesis steps of Example 2 refer to Example 1, wherein in the third step, compound Q5 is replaced by compound Q6 to synthesize compound I-2.

[0201] MS m / z(ESI):527[M+1].

[0202] HNMR: (400MHz, DMSO-d6)9.39(s,1H),8.56-8.55(m,1H),8.54(brs,2H),7.95 (s,1H),7.93-7.91(m,1H),7.40-7.34(m,2H),5.92-5.90(m,1H),4.67-4.65( m,1H),4.64(brs,1H),4.52-4.10(m,2H),3.70-3.68(m,1H),3.61-3.43(m,6H ),3.16-3.12(m,1H),2.15-2.12(m,1H),2.03-1.93(m,4H),1.71-1.53(m,3H).

[0203] Example 3: Preparation of the compound represented by formula I-3

[0204]

[0205] The synthesis steps of Example 3 refer to Example 1, wherein in the third step, compound Q5 is replaced by compound Q7 to synthesize compound I-3.

[0206] MS m / z(ESI):527[M+1].

[0207] HNMR: (400MHz, DMSO-d6)9.39(s,1H),8.56-8.55(m,1H),8.54(brs,2H),7.95 (s,1H),7.93-7.91(m,1H),7.40-7.34(m,2H),5.92-5.90(m,1H),4.67-4.65( m,1H),4.64(brs,1H),4.52-4.10(m,2H),3.70-3.68(m,1H),3.61-3.43(m,6H ),3.16-3.12(m,1H),2.15-2.12(m,1H),2.03-1.93(m,4H),1.71-1.53(m,3H).

[0208] Example 4: Preparation of the compound represented by formula I-4

[0209]

[0210] The synthesis steps of Example 4 refer to Example 1, wherein the intermediate compound Q2 is replaced by Q3 in the first step, and the compound Q5 is replaced by compound Q7 in the third step to synthesize compound I-4.

[0211] MS m / z(ESI):528[M+1].

[0212] HNMR: (400MHz, DMSO)9.39(s,1H),8.56-8.55(m,1H),8.54(brs,2H),7.95(s ,1H),7.93-7.91(m,1H),7.40-7.34(m,2H),5.92-5.90(m,1H),4.67-4.65(m, 1H),4.64(brs,1H),4.52-4.10(m,2H),3.70-3.68(m,1H),3.61-3.43(m,6H) ,3.16-3.12(m,1H),2.15-2.12(m,1H),2.03-1.93(m,4H),1.71-1.53(m,3H).

[0213] Example 5: Preparation of the compound represented by formula I-5

[0214]

[0215] The synthesis steps of Example 5 refer to Example 1, wherein in the third step, compound Q5 is replaced by compound Q8 to synthesize compound I-5.

[0216] MS m / z(ESI):528[M+1].

[0217] HNMR: (400MHz, DMSO)8.57-8.56(m,1H),8.44(brs,3H),8.12(s,1H),7.96-7.94(m, 1H),7.89-7.88(m,1H),7.69-7.68(m,1H),7.39-7.35(m,2H),6.05(d,J=6.4MHz,1H ),4.82-4.78(m,1H),4.55-4.54(m,1H),4.11-3.96(m,5H),3.78-3.73(m,3H),2.03 -1.93(m,4H).1.77-1.72(m,2H),1.64(m,1H),0.77-0.76(m,2H),0.70-0.67(m,2H).

[0218] Example 6: Preparation of the compound represented by formula I-6

[0219]

[0220] The synthesis steps of Example 6 refer to Example 1, wherein the intermediate compound Q2 is replaced by Q4 in the first step, and the compound Q5 is replaced by compound Q7 in the third step to synthesize compound I-6.

[0221] MS m / z(ESI):528[M+1].

[0222] HNMR: (400MHz, DMSO-d6)8.73(brs,3H),8.14(s,1H),8.56-8.55(m,2H),8.27(s,1H),8,06-8.05(m,1H),7.40-7.38(m,1H),7.33-7.31(m,1H) ,6.21-6.20(m,1H),5.07-4.98(m,2H),4.85-4.83(m,1H),4.52(m,1H) ,3.73-3.49(m,7H),2.17(m,2H).2.03-1.99(m,4H),1.74-1.43(m,4H).

[0223] Effect Example 1: Inhibition of Cell Proliferation Experiment

[0224] Experimental Materials:

[0225] RPMI-1640 medium and penicillin / streptomycin antibiotics were purchased from Vicente, and fetal bovine serum was purchased from Biosera. 3D CellTiter-Glo (a chemiluminescent cell viability assay) reagent was purchased from Promega. Staurosporine was purchased from Taosu Biochemical. NCI-H358 or MV-4-11 cell lines were purchased from Wuhan Punosai Life Science Co., Ltd. Nivo multi-label analyzer was purchased from PerkinElmer.

[0226] Experimental methods:

[0227] NCI-H358 cells or MV-4-11 cells were seeded into 96-well ultra-low attachment U-shaped plates, with 80 μL of cell suspension per well containing 2,000 NCI-H358 cells. The plates were incubated overnight in a CO2 incubator.

[0228] The test compound was diluted three-fold using a dispenser to the ninth concentration, from 600 μM to 100 nM, in duplicate. 78 μL of culture medium was added to the middle plate. 2 μL of the serially diluted compound was then transferred to each well of the middle plate, mixed thoroughly, and 20 μL was transferred to each well of the cell plate. The concentration of the compound transferred to the cell plate ranged from 3 μM to 0.5 nM. The cell plate was incubated in a CO2 incubator for 5 days.

[0229] On the day of reading, add 100 μL of cell viability chemiluminescent detection reagent to each well of the cell plate and incubate at room temperature for 10 minutes to allow the luminescent signal to stabilize. Read using a multi-label analyzer.

[0230] Max well: DMSO solvent well

[0231] Min wells: staurosporine-treated wells

[0232] Data Analysis:

[0233] The raw data were converted into inhibition rate, IC, using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by four-parameter curve fitting (obtained in the "log (inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Table 1 provides the inhibitory activity of the compounds of the present invention on the proliferation of NCI-H358 cells or MV-4-11 cells.

[0234] Table 1: Cell proliferation inhibition activity data of the compounds of the present invention (IC 50 )

[0235]

[0236] From the experimental results in Table 1, we can see that the example compounds of the present invention have good activity in inhibiting the proliferation of NCI-H358 or MV-4-11 cells. The activity of some compounds far exceeds that of the control, showing extremely important anti-tumor potential.

[0237] Effect Example 2: Drug Metabolism Experiment

[0238] 1) The compound of the present invention prepared in the above examples was prepared as a 0.3 mg / mL clear solution (2% DMSO + 30% PEG 300 + 2% Tween 80 + 66% H 2 O) for oral administration and a 0.2 mg / mL clear solution (2% DMSO + 30% PEG 300 + 2% Tween 80 + 66% H 2 O) for intravenous administration.

[0239] 2) Male CD-1 mice, 3 per group, weighing 27-28 g, were provided by Shanghai Slake Laboratory Animal Co., Ltd. The mice were given a 2-4 day acclimatization period before the experiment and fasted for 8-12 hours before dosing. Water was given 2 hours after dosing and food was given 4 hours after dosing.

[0240] 3) After the mice were fasted for 12 hours but had free access to water, blank plasma was collected at time 0;

[0241] 4) Take the mice prepared in step 2) and administer the test compound at 3 mg / kg orally (PO) or 1 mg / kg intravenously (IV);

[0242] 5) 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 10 h, and 24 h after oral administration, continuously draw blood from the retinal venous plexus and place it in heparinized EP tubes. Centrifuge at 8000 rpm / min for 5 min, then collect the upper plasma layer and freeze at -20°C for LC-MS / MS analysis.

[0243] 6) Based on the blood drug concentration-time data obtained in step 5), pharmacokinetic parameters were calculated using WinNonlin software. Specific data are shown in Table 2.

[0244] Table 2: Pharmacokinetic data of the compounds of the present invention

[0245]

[0246] Pharmacokinetic experimental data are shown in Table 2. The results indicate that oral or intravenous administration of some of the exemplary compounds of the present invention to mice resulted in very high exposure in animal plasma and excellent half-life, tissue distribution, area under the curve, and bioavailability. Some of the compounds performed better than the reference substance, TNO-155, and have promising prospects for clinical application.

[0247] Throughout this specification, references to terms such as "one embodiment," "some embodiments," "examples," "specific examples," or "some examples" indicate that the specific features, structures, materials, or characteristics described in conjunction with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0248] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention without departing from the principles and purpose of the present invention.

Claims

1. The following compound or a pharmaceutically acceptable salt thereof:

2. A pharmaceutical composition, characterized in that The pharmaceutical composition contains an effective dose of the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

3. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 2, in the preparation of a medicament for treating and / or preventing cancer, wherein the cancer is selected from acute myeloid leukemia and non-small cell lung cancer.

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