CN114163457A - Pyrimido five-membered nitrogen heterocyclic compound and use thereof - Google Patents

Abstract

The present invention relates to pyrimido five-membered nitrogen heterocyclic compounds and uses thereof. In particular, the present invention relates to a pyrimido five-membered nitrogen heterocyclic derivative represented by the general formula (I), its preparation method, a pharmaceutical composition containing the derivative, and its use as a SHP2 inhibitor for prevention and/or Use in the treatment of tumors or cancer, wherein each substituent in the general formula (I) is as defined in the specification.

Description

Pyrimido five-membered nitrogen heterocyclic compound and use thereof

technical field

The invention discloses pyrimido five-membered nitrogen heterocyclic compounds, pharmaceutically acceptable salts, hydrates, prodrugs, stereoisomers, solvates or isotopically labeled compounds thereof. The present invention also provides preparation methods of such compounds and intermediate compounds thereof, compositions containing such compounds, and use of such compounds in preparing medicines for preventing and/or treating diseases or conditions related to abnormal SHP2 activity.

Background technique

The tyrosine phosphatase SHP2 consists 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 can combine with PTP to form a ring structure, thereby hindering the binding of PTP to the substrate, so that the catalytic activity of the enzyme is inhibited; when the tyrosine of the upstream receptor protein is phosphorylated, NSH2 and In combination, the PTP catalytic domain is released to exert phosphatase activity.

At the cellular level, SHP2 participates in multiple tumor cell signaling pathways, such as RTK/Ras/MAPK, JAK/STAT, and PB3K/Akt, by functioning downstream of many receptor tyrosine kinases in the cytoplasm. Through the regulation of these kinases and signaling pathways, SHP2 is closely related to many important cell life activities, such as cell proliferation, migration, differentiation, death, regulation of cytokines, and tumorigenesis.

Meanwhile, SHP2 is also involved in programmed death receptor 1 (PD1)-mediated suppression of the immune system. After the PD-1 of T cells binds to PD-L1, a large amount of SHP2 can be recruited in the cell. SHP2 can dephosphorylate antigen receptor pathway proteins in T cells, thereby inhibiting T cell activation. Therefore, inhibition of SHP2 activity could reverse immunosuppression in the tumor microenvironment.

As an important cell signaling factor, SHP-2 mutation is closely related to a variety of diseases. The study found that SHP- 2 mutations.

At present, several allosteric inhibitors of SHP-2 have entered the clinical research stage, such as TNO-155 developed by Novartis, RMC-4630 developed by Revolution Medicine, and JAB-3068 developed by Beijing Jiakesi. clinical research stage. However, there is no SHP-2 inhibitor developed and marketed for the preparation of Noonan syndrome, Leopard skin syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, head and neck cancer, lung cancer and other Colon cancer. Therefore, there is an urgent need to develop a class of SHIP-2 inhibitors with good druggability.

SUMMARY OF THE INVENTION

The pyrimidine five-membered nitrogen heterocyclic compound provided by the present invention is a new class of SHP2 inhibitors, exhibits good inhibitory activity on tumor cells and good druggability, and has broad drug development prospects. Moreover, the preparation method of the compound is simple, which is favorable for industrial production.

According to the first aspect of the present invention, the present invention provides a compound shown in formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer, or a hydrate thereof , or its solvate, or its metabolite, or its prodrug,

in,

X ¹ , X ² or X ³ are independently selected from CR ⁵ or N, and at least one of them is N;

X ⁴ or X ⁵ is independently selected from C or N;

R ¹ , R ² , R ³ and R ⁴ are independently selected from H, halogen, hydroxy, amino, cyano, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₁ -C ₈ aldehyde, C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₈ alkoxy or C ₁ -C ₃ Haloalkoxy, the C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₈ alkoxy Or C ₁ -C ₃ haloalkoxy is optionally substituted by one or more R ⁵ , when substituted by multiple R ⁵ , R ⁵ may be the same or different;

R ⁵ is selected from H, halogen, hydroxyl, amino, cyano, C ₁ -C ₃ alkyl, C ₁ -C ₃ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl , C ₁ -C ₃ alkoxy or C ₁ -C ₃ haloalkoxy;

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

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

Ring A is independently selected from C ₃ -C ₈ cycloalkyl, C ₆ -C ₁₂ spirocycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl or C ₅ -C ₁₂ heteroaryl base;

Ring B is independently selected from C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl or C ₅ -C ₁₂ heteroaryl;

The heteroatom or heteroatom group contained in the heteroalkyl group, heterocycloalkyl group and heteroaryl group is 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 said heteroatoms or heteroatomic groups is independently selected from 1, 2 and 3, respectively.

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by the formula (I) according to the present invention, when X ¹ and X ³ are N, X ² is C.

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by the formula (I) according to the present invention, when X ² and X ³ are N, X ¹ is C.

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by the formula (I) according to the present invention, when X ¹ and X ² are C, X ³ is N.

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by the formula (I) described in the present invention, when X ⁴ is N, ring A is selected from pyrrole ring, spirocyclic pyrrole ring, Piperidine ring, spirocyclic piperidine ring or dihydropyrazole ring.

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by the formula (I) according to the present invention, when X ⁴ is N, the ring A is independently selected from the group consisting of

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

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

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

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) according to the present invention, when X ⁵ is N, ring B is independently

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

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by formula (I) according to the present invention, when X ⁵ is C, ring B is independently selected from the group consisting of

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by the formula (I) of the present invention, R ¹ , R ² , R ³ and R ⁴ are selected from H, halogen, hydroxyl, Amino, cyano, C ₂ -C ₄ alkenyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ heteroalkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₃ haloalkoxy, the C ₁ -C ₄ alkyl, C ₁ -C ₄ heteroalkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ Heterocycloalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₃ haloalkoxy is optionally substituted by one or more R ⁵ , when substituted by multiple R ⁵ , R ⁵ may be the same or different; wherein R ⁵ is selected from H, F, Cl, Br, hydroxy, amino, cyano, methyl, ethyl, cyclopropyl, methoxy, ethoxy, trifluoromethoxy.

In one of the preferred examples, in the structure of the pyrimido five-membered nitrogen heterocyclic compound represented by the formula (I) of the present invention, R ¹ , R ² , R ³ and R ⁴ are selected from H, halogen, hydroxyl, Amino, cyano, methyl, ethyl, isopropyl, cyclopropyl, vinyl, methoxy, ethoxy, isopropoxy, trifluoromethyl, difluoromethyl, trifluoromethoxy , -OCH ₂ CHF ₂ , -N(CH ₃ ) ₂ , -NH(CH ₃ ) or -OCH ₂ CF ₃ .

In some embodiments, the compound of formula (I) described in the present invention is selected from the following compounds of formula (II), formula (III) or formula (IV):

wherein 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.

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

wherein 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.

According to a specific embodiment of the present invention, the compound shown in formula I of the present invention is any of the following compounds:

In the present invention, those skilled in the art can select the group and its substituent in the compound shown in formula I to provide stable compound shown in formula I, or a pharmaceutically acceptable salt thereof, or its stereoisomer, or its tautomer, or its hydrate, or its solvate, or its metabolite, or its prodrug, including but not limited to the I- 1 to I-6.

The reaction solvent used in each reaction step of the present invention is not particularly limited, and 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 in the art, equivalent replacements, or equivalents to the solvents, solvent combinations, and different ratios of solvent combinations described in the present invention are all deemed to be within the scope of the present invention.

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

The pharmaceutical excipients can be those widely used in the field of pharmaceutical production. Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide a method to enable the subject to dissolve the active ingredient at a desired rate after administration, or to facilitate the subject to receive the composition after administration. Active ingredients are effectively absorbed. The pharmaceutical excipients can be inert fillers, or provide some function, such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. Described pharmaceutical adjuvants may include one or more of the following adjuvants: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrating agents, lubricants, anti-sticking agents Agents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavors and sweeteners.

The pharmaceutical compositions of the present invention can be prepared in light of the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, attenuating, encapsulating, entrapping or lyophilizing processes.

The pharmaceutical compositions of the present invention may be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ophthalmic, rectal, topical or parenteral (infusion, injection, implant Intradermal, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical compositions of the present invention may also be in controlled release or delayed release dosage forms (eg, liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, caplets, softgels, 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, creams, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum formulations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry formulations 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 compositions include, but are not limited to, eye drops and other ophthalmic formulations; aerosols: such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories and lozenges agent.

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 situation, other modes of administration may be applied or even preferred. For example, transdermal administration may be highly desirable in patients who are forgetful or irritable with oral medications. In particular cases, the compounds of the present invention may also be administered by transdermal, intramuscular, intranasal or intrarectal routes. The route of administration may vary in any manner limited by the physical properties of the drug, convenience of the patient and caregiver, and other relevant circumstances.

According to the third aspect of the present invention, the present invention provides a compound represented by formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer or a tautomer thereof, or a pharmaceutical composition thereof in Use in the preparation of a medicament for treating and/or preventing diseases caused by abnormal mutation of SHP2. The compounds provided by the present invention can be used for the treatment and/or prevention of one or more diseases caused by abnormal mutation of SHP2, and have good clinical and medical applications.

According to the fourth aspect of the present invention, the present invention provides a compound represented by formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate thereof, Use of its solvate, its metabolite, or its prodrug, or its pharmaceutical composition in the preparation of SHP2 inhibitor medicines. The compounds provided by the present invention have excellent SHP2 enzymatic activity and cell proliferation inhibitory activity, and can be effectively used as SHP2 inhibitors and as therapeutic drugs for SHP2 inhibitors.

According to the fifth aspect of the present invention, the present invention provides a compound shown in formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate thereof, Use of its solvate, or its metabolite, or its prodrug, or its pharmaceutical composition in the preparation of a drug for treating and/or preventing cancer. The compounds provided by the present invention can be used to prepare medicines for the treatment and/or prevention of cancer, wherein the cancers include but are not limited to Noonan syndrome, Leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, Melanoma, acute myeloid leukemia, breast cancer, esophageal cancer, lung cancer, colon cancer, stomach cancer, anaplastic large cell lymphoma, glioblastoma, non-small cell lung cancer, or head and neck tumors.

The present invention has the following advantages:

1. The pyrimido five-membered nitrogen heterocyclic compound disclosed in the present invention is a new type of allosteric inhibitor, which can inhibit the SHP2 by combining with the non-catalytic region of SHP2 and "locking" the weak basic state of SHP2 activity. active purpose. The pyrimido five-membered nitrogen heterocyclic compound disclosed by the invention overcomes the disadvantages of general selectivity and poor druggability of PTP catalytic region inhibitors, exhibits good biological activity and druggability, and has good drug development prospects.

2. In the evaluation system of SHP2 enzyme activity inhibition experiment, phosphorylated protein kinase (p-ERK) cell experiment, NCI-H358 cell, MV-4-11 cell proliferation inhibition experiment and other evaluation systems under the same conditions, the present invention is different from the one with the disclosed structure. Compared with RMC4550 and TNO155, compound RMC4550 showed superior activity and PK properties.

Terms and Definitions

Unless stated to the contrary, the following terms used in the specification and claims have the following meanings.

The term "alkyl" refers to saturated aliphatic hydrocarbon groups, eg, straight and branched chain groups comprising 1 to 20 carbon atoms, eg, may be 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 20 carbon atoms Straight and branched chain groups of 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. As used herein, "alkyl" may 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 isomers thereof, etc. Non-limiting examples also include methylene, methine, ethylene, ethylene, propylene, propylene, butylene, butylene, and various branched chain isomers thereof. Alkyl groups can be optionally substituted or unsubstituted.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, eg, comprising 3 to 12 ring atoms, such as may be 3 to 12, 3 to 10, or 3 to 12 6 ring atoms, or can be a 3, 4, 5, 6 membered ring. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptyl Alkenyl, cyclooctyl, etc. Cyclic groups can be optionally substituted or unsubstituted.

The term "spirocycloalkyl" refers to a cycloalkyl group in which two rings share a single ring atom. Shared ring atoms are also referred to as spiro atoms, most often referred to as quaternary carbons (spiro carbons).

The term "heteroalkyl" refers to a stable straight or branched chain consisting of the specified number of carbon atoms and one or more heteroatoms selected from the group consisting of O, N, and S, and wherein the nitrogen and sulfur atoms are optionally is 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 -CH ₂ -CH ₃ , -CH ₂ -CH ₂ -S(O)-CH ₃ , -CH ₂ -CH ₂ -S(O) ₂ -CH ₃ or -CH ₂ -CH=N-OCH ₃ . Up to two heteroatoms may be consecutive, eg _-CH2 -NH-O- _CH3 . In certain embodiments, heteroalkyl groups are optionally substituted as described elsewhere herein.

The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, eg, comprising 3 to 20 ring atoms, eg, may be 3 to 16, 3 to 12, 3 to 12 10 or 3 to 6 ring atoms, one or more of which is selected from nitrogen, oxygen, or a heteroatom of S(O) _m (where m is 0, 1, or 2), excluding -OO-, The ring moiety of -OS- or -SS-, the remaining ring atoms being carbon. In some embodiments, it is preferred to include 3 to 12 ring atoms, of which 1-4 are heteroatoms, more preferably the heterocycloalkyl ring includes 3 to 10 ring atoms, most preferably a 5-membered ring or a 6-membered ring, wherein 1-4 are heteroatoms, more preferably 1-3 are heteroatoms, and most preferably 1-2 are heteroatoms. Non-limiting examples of monocyclic heterocycloalkyl groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Non-limiting examples of polycyclic heterocycloalkyl groups include spirocyclic, fused ring, or bridged ring heterocycloalkyl groups.

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

The term "alkynyl" refers to a straight or branched chain unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms 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≡CC≡CH) and the like.

The term "alkoxy" refers to -O-alkyl.

The term "haloalkoxy" means that an alkoxy group is substituted with one or more halogen atoms, the same or different, such as trifluoromethoxy and the like.

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated pi 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-tetralin, and the like.

The term "heteroaryl" refers to the group remaining after the removal of one hydrogen atom from the "heteroaromatic ring" molecule. Heteroaryl may be unsubstituted or substituted. The substituents include but are not limited to alkyl, alkyl Oxy, aryl, aralkyl, amino, halogen, hydroxy, cyano, nitro, carbonyl and heteroalicyclic. Non-limiting examples of unsubstituted heteroaryl groups include, but are not limited to, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolyl, iso Quinolinyl, tetrazolyl, triazinyl.

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

The term "amino" refers to _-NH2 groups, -NH(alkyl) and -N(alkyl) ₂ , specific examples of amino groups include but are not limited to _-NH2 , -NHCH3, -NHCH( _{CH3 )2 ,} -N(CH ₃ ) ₂ , -NHC ₂ H ₅ , -N(CH ₃ )C ₂ H ₅ and the like.

The term "hydroxy" refers to the -OH group.

The term "cyano" refers to the -CN group.

The terms "optional" or "optionally" mean that the subsequently described event or circumstance may or may not occur. For example: "heterocyclic group optionally substituted by alkyl" means that the alkyl group may or may not be present, that is, including the case where the heterocyclic group is substituted by the alkyl group and the case where the heterocyclic group is not substituted by the alkyl group .

"Substituted" refers to the replacement of one or more hydrogen atoms in a group with a substituent. For example, up to 5, more preferably 1 to 3, hydrogen atoms in the group are substituted independently of each other by the corresponding number of substituents.

"Pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic 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 forms 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 ammonia 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 including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts including, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, and methanesulfonic acids; also include salts of amino acids such as arginine, etc. , and salts of 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 thus can be converted into either base or acid addition salts. Preferably, the neutral form of the compound is regenerated by contacting the salt with a base or acid in a conventional manner and isolating the parent compound. The parent form of a compound differs from its various salt forms by certain physical properties, such as solubility in polar solvents. According to an embodiment of the present invention, preferably the pharmaceutically acceptable salt of the compound represented by formula I of the present invention is hydrochloride, hydrobromide, phosphate, or sulfate, most preferably hydrochloride.

The term "solvate" refers to a physical association of a compound of the present invention with one or more solvent molecules. This physical association includes various degrees of ionic and covalent bonding, including hydrogen bonding. "Solvate" includes solution phases and isolatable solvates. Non-limiting examples of solvates include ethanolates, methanolates, and the like. A "hydrate" is a solvate in which the solvent molecule is _H2O .

"Pharmaceutical composition" means a mixture containing one or more compounds, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, and other chemical components, as well as other components such as Pharmaceutically acceptable excipients. The purpose of the pharmaceutical composition is to facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.

Any formula or structure given herein is also intended to represent unlabeled as well as isotopically labeled forms of the compounds. Isotopically-labeled compounds have the structures shown by the formulae 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 of the present disclosure, such as those into which radioactive isotopes such ^as3H , ^13C , ^and14C have been incorporated. Such isotopically labeled compounds can be used in metabolic studies, reaction kinetic studies, detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography including drug or substrate tissue distribution analysis (SPECT), or radiotherapy for patients. Isotopically-labeled compounds of the present disclosure and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the Schemes or in the Examples and Preparations described below, by substituting a readily available isotopically-labeled reagent for a non-isotopically-labeled reagent.

The compounds of the present invention may be in the form of prodrug compounds. "Prodrug compound" means a derivative converted into a compound according to the invention by reaction with an enzyme or gastric acid etc. under physiological conditions in an organism, for example by oxidation, reduction or hydrolysis etc., each of said reactions species are carried out enzymatically. Examples of prodrugs are compounds wherein an amino group in a compound of the invention is acylated, alkylated or phosphorylated to form, for example, eicosanoylamino, alanylamino, pivaloyloxymethylamino, or Where hydroxyl groups are acylated, alkylated, phosphorylated or converted to boronate esters such as acetyloxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaryloxy, alanyloxy oxy, or in which the carboxyl group is esterified or amidated. These compounds can be produced from the compounds of the present invention according to known methods. Other examples of prodrugs are compounds wherein, for example, the carboxylate in the compounds of the invention is converted to an alkyl-, aryl-, choline-, amino, acyloxymethyl ester, linolenyl ester.

Where tautomerism such as, for example, keto-enol tautomerism, of the compounds of the invention or their prodrugs would occur, individual forms such as, for example, keto and enol forms and their in any ratio mixtures of each are within the scope of the present invention. The same applies to stereoisomers such as, for example, enantiomers, cis/trans isomers, conformation isomers, and the like.

If desired, the isomers can be separated by methods well known in the art, such as by liquid chromatography. The same applies to enantiomers by using eg chiral stationary phases. Furthermore, enantiomers can be separated by converting them to diastereomers by coupling with an enantiomerically pure auxiliary compound, followed by separation of the resulting diastereomers and cleavage of the auxiliary residue base. Alternatively, any enantiomer of the compounds of the present invention can be obtained from stereoselective synthesis using optically pure starting materials.

The term "treating" means administering a compound or formulation of the invention to prevent, ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:

(i) preventing the emergence of a disease or disease state in mammals, particularly when such mammals are susceptible to, but have not been diagnosed with, the disease state;

(ii) inhibiting the disease or disease state, i.e. arresting its development;

(iii) Alleviation of the disease or disease state, even if the disease or disease state resolves.

The term "effective dose" means (i) treatment or prevention of a particular disease, condition or disorder, (ii) alleviation, amelioration or elimination of one or more symptoms of a particular disease, condition or disorder, or (iii) prevention or delay herein The amount of a compound of the present invention to be used for the onset of one or more symptoms of a particular disease, condition or disorder described in . The amount of a compound of the invention that constitutes an "effective dose" will vary 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 of ordinary skill in the art according to the own knowledge and this disclosure.

Additional aspects and advantages of the present invention will be set forth, in part, from the following description, and in part will be apparent from the following description, or may be learned by practice of the invention.

The specific reaction conditions are described as follows:

Compound A and compound B are heated and reacted in DMF, DMSO, NMP and other solvents under basic conditions such as potassium carbonate, DIEA, cesium carbonate, sodium tert-butoxide, etc. to obtain compound C. Compound C and Boc acid anhydride are in methylene chloride, obtain compound D with DMAP catalysis, compound D and compound E make catalyst through Pd ₂ (dba) ₃ , take Xantphos etc. as ligand, add organic base or inorganic base, such as DIEA, cesium carbonate, etc., to obtain compound F, and compound F is deprotected with TFA or ethyl acetate hydrochloride to obtain the compound represented by general formula I in the form of free or hydrochloride.

Detailed ways

The compound represented by the formula I of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate, or a solvate thereof, or a metabolite thereof, The preparation of or its prodrugs can be accomplished by the exemplary methods described in the following examples and the relevant published literature operations used by those skilled in the art, but these examples do not limit the scope of the present invention.

The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR was measured using Bruker AVANCE-400 or Varian Oxford-300 nuclear magnetometer, and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDC1 ₃ ), deuterated methanol (CD ₃ OD), The internal standard is tetramethylsilane (TMS) and chemical shifts are given in units of ^10-6 (ppm).

The 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).

The determination of HPLC used an Agilent 1200DAD high pressure liquid chromatograph (Sunfirc C18, 150X4.6mm, 5wn, column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18, 150X4.6mm, 5μm column).

The thin layer chromatography silica gel plate uses Qingdao Ocean GF254 silica gel plate, the size of the silica gel plate used for thin layer chromatography (TLC) is 0.15mm-0.2mm, and the size of the thin layer chromatography separation and purification products is 0.4mm-0.5mm silicone board.

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

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

Unless otherwise specified in the examples, the reactions were all carried out in an argon atmosphere or a nitrogen atmosphere. The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times.

Unless otherwise specified in the examples, the reaction temperature is room temperature, and the temperature range is 5°C-35°C.

The monitoring of the reaction progress in the embodiment adopts thin layer chromatography (TLC), and the system of the developing agent used in the reaction has A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system, and the volume ratio of the solvent is according to The polarity of the compounds can be adjusted.

The eluent system for column chromatography and the developing solvent system for thin layer chromatography used to purify the compound include A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system, and the volume ratio of the solvent is based on the compound Different polarities can be adjusted, and a small amount of triethylamine and acidic or basic reagents can also be added for adjustment.

The present invention will be described in detail by the following examples, but it does not mean any unfavorable limitation of 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 enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, 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 invention. The following synthetic schemes describe the steps for the preparation of the compounds disclosed herein. Unless otherwise stated, each substituent has the meaning as described herein.

Synthesis of Intermediates:

Synthesis of Intermediate Q1

Step 1: Synthesize Q1-2

Compound Q1-1 (100g, 495mmol) was added to DME (1000ml), cooled to -10°C and then added isobutyl chloroformate (67.6g, 495mmol) and 4-methylmorpholine (50g, 495mmol), and the reaction was carried out at room temperature 5 hours, after the TLC reaction finished, filter, filter and wash the solid with DME (250ml), then add the filtrate into a 2L there-necked flask, add sodium borohydride (37.6g, 990mmol) to process the filtrate, stir at room temperature for 30min, then slowly dropwise Add methanol (250ml), continue to react at room temperature for 3 hours, the TLC reaction is completed, the reaction solution is spin-dried, water (1500ml) is added, the aqueous phase is extracted with DCM (300ml*3), after the organic phases are combined, the organic phases are followed by water and After washing with saturated sodium chloride, the organic phase was dried and spin-dried to obtain compound Q1-2 (86.5 g, white solid) with a yield of 93%. used directly in the next step.

Step 2: Synthesize Q1-3

Compound Q1-2 (25 g, 133 mmol)) was added to DCM (250 ml), then TEA (26.9 g, 266 mmol) was added, and MsCl (18.3 g, 156 mmol) was added dropwise under ice bath, and then reacted at room temperature for 1 h. After TLC showed that the reaction was over, the reaction solution was diluted with DCM, washed with water and saturated sodium chloride successively, and then the organic phase was dried, concentrated to dryness, and the obtained residue was subjected to silica gel chromatography (eluent petroleum ether). : ethyl acetate=3:1～1:1) to obtain compound Q1-3 (33.6 g, colorless transparent liquid) in a yield of 95%.

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

Step 3: Synthesize Q1-4

N-Boc-4-cyanopiperidine (28g, 130mmol) was added to THF (300ml), cooled to -78°C, 2.0M LDA (75ml, 150mmol) was slowly added dropwise, after the dropwise addition, continued to maintain - The reaction was carried out at 78°C for 1.5h, and then the THF solution (150ml) of compound Q1-3 (26.6, 100mmol) obtained in one step was added dropwise. After the addition was completed, the reaction was kept at -78°C for 3h. Saturated ammonium chloride solution (50ml) was used to quench the reaction, and saturated aqueous sodium chloride solution (500ml) was added, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (150ml*3), the organic phases were combined, and the organic Dry over anhydrous sodium sulfate, filter, spin dry and concentrate the obtained residue and pass through silica gel column chromatography (eluent petroleum ether: ethyl acetate=10:1～1:1) to obtain compound Q1-4 (25.7g, white solid) ) in 67.6% yield.

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

Step 4: Synthesize Q1-5

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

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

Step 5: Synthesis of Q1-6

The compound Q1-5 (10g, 33mol) obtained in the previous step was added to tetraethyl titanate (100ml), then (R)-(+)-tert-butylsulfinamide (4.8g, 40mml) was added, and the temperature was increased. The reaction was carried out at 90 °C for 3 hours. After TLC showed that the reaction was completed, it was cooled to room temperature. The reaction solution was slowly added to ice water (500 ml), and the obtained aqueous phase was extracted with dichloromethane (150 mL×3). The phase was washed with saturated brine (100 mL×2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the residue obtained by spin-drying and concentration was passed 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%.

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

Step 6: Synthesis of Q1-7

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

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

Step 7: Synthesis of Intermediate Q1

The compound Q1-7 (5g, 12.2mmol) obtained in the previous step was added to ethyl acetate (50ml), then 4M HCl/ethyl acetate solution (25ml) was added, and the reaction was carried out at room temperature for 1h. After TLC showed that the reaction was completed, After filtration, 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%.

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

Synthesis of Intermediate Q2

Step 1: Synthesize Q2-2

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

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

Step 2: Synthesis of Q2-3

Compound Q2-2 (5.4 g, 18.2 mmol) obtained in the previous step was dissolved in concentrated sulfuric acid (50 mL), and then the temperature was raised to 75°C. After 2 hours of reaction, TLC showed that the reaction was completed. After cooling to room temperature, the reaction solution was slowly poured into ice water, and then slowly alkalized to about pH 6 with 5M aqueous sodium hydroxide solution, a large amount of solid was precipitated, filtered, and the filter cake was washed with water, The solid was dried to give compound Q2-3 (3.2 g, grey solid) in 82.5% yield.

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

The third step: synthesis of intermediate Q2

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

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

Synthesis of Intermediate Q3

Step 1: Synthesis of Q3-2

5-Iodo-2,4-dichloropyrimidine (16.3 g, 59.2 mmol) and hydrazine hydrate (8.8 mL, 181 mmol) were added to absolute ethanol (300 mL), followed by heating under reflux for 12 hours. TLC showed that after the reaction was completed, it was cooled to 0°C, a large amount of solid was precipitated, filtered, and the filter cake was washed with iced ethanol (100 mL) and dried to obtain Q3-2 (15.1 g, yellow solid) in a yield of 94.3%, which did not require Further purification was used for the next step.

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

Step 2: Synthesis of Intermediate Q3

The compound Q3-2 (15g, 55.3mmol) obtained in the previous step was added to trimethyl orthoformate (100ml), heated to reflux for 12 hours, TLC showed that the reaction was over, cooled to 0°C, a large amount of solid was precipitated, filtered, and filtered. The cake was washed with iced 50% ethanol (50 mL) and dried to give compound Q3 (11.3 g, yellow solid) in 88% yield, which was used in the next step without further purification.

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

Synthesis of Intermediate Q4

Step 1: Synthesis of compound Q4-2

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

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

Step 2: Synthesis of compound Q4-3

Compound Q4-2 (42 g, 160 mmol) obtained in the previous step was added to methanol (250 mL), followed by 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 completed, the reaction solution was cooled to 0 °C, a large amount of solid was precipitated, filtered, and the filter cake was washed with 100 mL of iced methanol, and the solid was dried to obtain compound Q4-3. (33.6 g, white solid) in 84% yield, which was used in the next step without further purification.

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

The third step: synthesis of compound Q4-4

The compound Q4-3 (7.5g, 29.6mmol) obtained in the previous step was added to polyphosphoric acid (50ml), the reaction temperature was raised to 120°C, and the reaction was stirred for 5 hours. 150mL ice water, adjust the pH to about 8 with 2N aqueous sodium hydroxide solution, then extract the obtained aqueous phase with n-butanol (150mL*3), combine the organic phases, and wash the combined organic phases with saturated brine (70mL× 2), and then the organic phase was dried with anhydrous sodium sulfate, filtered, and concentrated by spin drying to obtain compound Q4-4 (3.5 g, pale yellow solid) with a yield of 50%.

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

Step 4: Synthesis of Intermediate Q4

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

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

Synthesis of Intermediate Q5

Step 1: Synthesis of compound Q5-2

Compound Q5-1 (10 g, 88.4 mmol) was added to THF solution (100 ml), followed by addition of 2-hydroxymethylpyrrolidine-1-carboxylate butyl ester (21.4 g, 106.1 mmol, CAS: 170491-63- 1) and triphenylphosphine (34,8g, 132.6mmol), then the reaction solution was cooled to below 5°C with an ice-water bath, and then DEAD (23.1g, 132.6mmol) was slowly added dropwise. After the completion of the dropwise addition and stirring at room temperature for 16 hours, TLC indicated that the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue obtained by spin-drying was passed through a flash silica gel column (eluent petroleum ether: ethyl acetate = 10:1) to obtain compound Q5-2 (25 g, pale yellow oil) in a yield of 95.4%.

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

Step 2: Synthesis of compound Q5-3

The compound Q5-2 (5g, 16.8mmol) obtained in the previous step was added to ethyl acetate (50ml), and then 4M HCl/ethyl acetate solution (25ml) was added, and the reaction was carried out at room temperature for 1h. After TLC showed that the reaction was completed, filter , 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%.

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

The third step: synthesis of compound Q5-4

The hydrochloride (2.20 g, 8.17 mmol) of compound Q5-3 obtained in the previous step was added to ethanol (50 mL), then potassium carbonate (5.64 g, 40.87) was added, and after stirring at room temperature for half an hour, the temperature was increased. Stir at 65°C for 12h. TLC showed that the reaction was completed, then the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue obtained by spin-drying and concentration was passed through a flash silica gel column (eluent petroleum ether:ethyl acetate=10:1) to obtain compound Q5- 4 (1.22 g, colorless oil), 84.7% yield.

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

Step 4: Synthesis of compound Q5-5

Compound Q5-4 (1.2 g, 6.8 mmol) obtained in the previous step was dissolved in THF (20 ml), then cooled to -78 °C, and under the protection of _N2 atmosphere, n-butyllithium solution (5.4 mL, 13.6mmol, 2.5M hexane solution), after the dropwise addition, the temperature was naturally raised to 0°C and stirred for 1.5h. The reaction 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 above mixture. After the dropwise addition, the reaction solution was naturally warmed to room temperature and stirred at room temperature for 2 hours. After TLC showed that the reaction was completed, saturated aqueous NH ₄ Cl solution (100 ml) was added to the reaction solution, the obtained aqueous phase was extracted with ethyl acetate (100 mL×3), the organic phases were combined, and the organic phases were washed with saturated brine (70 mL×2 ), then the organic phase was dried with anhydrous sodium sulfate, filtered, and the residue obtained by spin-drying and concentration was passed through a silica gel chromatography column (eluent petroleum ether: ethyl acetate=3:1～1:1) to obtain compound Q5- 5 (1.5 g, yellow solid), 72.8% yield.

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

Step 5: Synthesis of compound Q5-6

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 ₂ (dba) ₃ (22.73 mg) , 0.025 mmol, 0.05 equiv) and Xantphos (14.36 mg, 0.025 mmol) and DIEA (387 mg, 3 mmol). After warming to 90°C under nitrogen and stirring for 1 hour, the reaction mixture was concentrated under reduced pressure. The residue obtained by spin-drying and concentration was passed through a silica gel chromatography column (eluent petroleum ether: ethyl acetate=3:1-1:1) to obtain compound Q5-6 (239 mg, pale yellow solid) in 81% yield.

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

Step 6: Synthesis of Intermediate Q5

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 hours, TLC showed that the reaction was complete, the reaction mixture was diluted with PE, and a large amount of solid was precipitated. The precipitated solid was collected by filtration, and washed with ethyl acetate, The intermediate (164 mg, pale yellow solid) was obtained in 84% yield.

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

Referring to the synthesis of intermediate Q5, using commercially available raw materials instead of butyl 2-hydroxymethylpyrrolidine-1-carboxylate, the following intermediates can be synthesized:

Example 1: Preparation of compound represented by formula I-1

synthetic route:

Step 1: Synthesis of Compound 1A

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 1.5 hours. After the reaction, 20 mL of ethyl acetate and 40 mL of water were added, extracted with ethyl acetate (20 mL*3), the organic phases were combined, washed with saturated sodium oxide solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. It was concentrated under pressure, and the residue was passed through a silica gel chromatography column (eluent dichloromethane:methanol=50:1-10:1) to obtain compound 1A (244 mg, pale yellow solid) in 64.3% yield.

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

Step 2: Synthesis of Compound 1B

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

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

Step 3: Synthesis of Compound 1C

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), Pd ₂ (dba) ₃ (22.73 mg, 0.025 mmol) and Xantphos ( 14.36 mg, 0.025 mmol) and DIEA (190 mg, 1.5 mmol). After warming to 90°C under nitrogen and stirring for 1 hour, the reaction mixture was concentrated under reduced pressure. The residue obtained by spin-drying and concentration was passed through a silica gel chromatography column (eluent dichloromethane:methanol=50:1-10:1) to obtain compound 1C (241 mg, pale yellow solid) in 77% yield.

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

Step 4: Synthesis of Compound I-1

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

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

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).

Example 2: Preparation of compound represented by formula I-2

For the synthesis steps of Example 2, refer to Example 1, wherein in the third step, compound Q5 is replaced by compound Q6, and compound I-2 is synthesized.

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

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).

Example 3: Preparation of compound represented by formula I-3

For the synthesis steps of Example 3, refer to Example 1, wherein in the third step, compound Q5 is replaced by compound Q7, and compound I-3 is synthesized.

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

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).

Example 4: Preparation of compound represented by formula I-4

For the synthesis steps of Example 4, refer to Example 1, wherein in the first step, the intermediate compound Q2 is replaced by Q3, and the third step is replaced by the compound Q7, and the compound I-4 is synthesized.

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

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).

Example 5: Preparation of compound represented by formula I-5

Refer to Example 1 for the synthesis steps of Example 5, wherein in the third step, compound Q5 is replaced by compound Q8, and compound I-5 is synthesized.

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

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).

Example 6: Preparation of compound represented by formula I-6

For the synthesis steps of Example 6, refer to Example 1, wherein in the first step, the intermediate compound Q2 is replaced by Q4, and the third step is replaced by the compound Q7 to replace the compound Q5, and the compound I-6 is synthesized.

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

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).

Effect Example 1: Inhibition of Cell Proliferation Experiment

Experimental Materials:

RPMI-1640 medium, penicillin/streptomycin antibiotics were purchased from Vicente, and fetal bovine serum was purchased from Biosera. 3D CellTiter-Glo (Cell Viability Chemiluminescence Detection Reagent) reagent was purchased from Promega. Staurosporine was purchased from Ceramic Biochemical. NCI-H358 cell line or MV-4-11 cell line was purchased from Wuhan Proceeds Life Technology Co., Ltd. Nivo Multilabel Analyzer (PerkinElmer).

experimental method:

NCI-H358 cells or MV-4-11 cells were seeded in a 96-well ultra-low adsorption U-shaped plate, 80 μL of cell suspension per well, which contained 2000 NCI-H358 cells. Cell plates were incubated overnight in a carbon dioxide incubator.

The compound to be tested was diluted 3-fold to the ninth concentration, that is, from 600 μM to 100 nM, and a double-well experiment was set up. Add 78 μL of medium to the middle plate, and then transfer 2 μL of each well of the compound to the middle plate according to the corresponding position. After mixing, transfer 20 μL of each well to the cell plate. The concentration of compounds transferred to the cell plate ranged from 3 [mu]M to 0.5 nM. The cell plates were placed in a carbon dioxide incubator for 5 days.

On the day of reading, 100 μL of cell viability chemiluminescence detection reagent was added to the cell plate and incubated at room temperature for 10 minutes to stabilize the luminescence signal. Read using a multi-label analyzer.

Max hole: DMSO solvent hole

Min well: Staurosporine treated well

data analysis:

Using the equation (Sample-Min)/(Max-Min)*100% to convert the raw data into inhibition rate, the IC ₅₀ value can be obtained by curve fitting with four parameters (“log(inhibitor) vs. response--Variable slope" mode). 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.

Table 1: Cell proliferation inhibitory activity data (IC ₅₀ ) of the compounds of the examples of the present invention

From the experimental results in Table 1, we can see that the example compounds shown in 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 reference substance, showing extremely important anti-tumor prospects.

Effect Example 2: Drug Metabolism Experiment

1) Using the compound of the present invention prepared in the above example, the oral drug was formulated into a 0.3 mg/mL clear solution (2% DMSO+30% PEG300+2% Tween 80+66% H ₂ O), and the intravenous drug was formulated into 0.2 mg /mL clear solution (2% DMSO + 30% PEG 300 + 2% Tween 80 + 66% _H2O ).

2) Male CD-1 mice, 3 mice in each group, weighing 27-28 g, provided by Shanghai Slack Laboratory Animal Co., Ltd. The tested mice were given a 2-4 day environmental adaptation period before the experiment, fasted for 8-12 hours before administration, given water 2 hours after administration, and given food after 4 hours.

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

4) Take the mice in step 2), and administer 3 mg/kg of the compound to be tested orally (PO); 1 mg/kg of the compound to be tested is administered intravenously (IV);

5) At 5min, 15min, 30min, 1h, 2h, 4h, 8h, 10h, 24h after oral administration, blood was continuously collected from the fundus venous plexus and placed in an EP tube distributed with heparin, and the upper plasma was collected after centrifugation at 8000rpm/min for 5min. Freeze at -20°C until LC-MS/MS analysis;

6) According to the blood drug concentration-time data obtained in step 5), use WinNonlin software to calculate the pharmacokinetic parameters, and the specific data are shown in Table 2.

Table 2: Pharmacokinetic data of the compounds of the examples of the present invention

Pharmacokinetic experimental data are shown in Table 2. The results show that after oral or intravenous administration of some of the compounds of the present invention to mice, there are very high exposures in animal plasma, very good half-life, tissue distribution, In terms of area under the curve and bioavailability, some compounds are better than the reference substance TNO-155, and have good clinical application prospects.

In the description of this specification, reference to the terms "one embodiment," "some embodiments," "example," "specific example," or "some examples", etc., means a specific feature described in connection with the embodiment or example, A structure, material, or feature is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

Although the embodiments of the present invention have been shown and described above, it should be understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and those of ordinary skill in the art will not depart from the principles and spirit of the present invention Variations, modifications, substitutions, and alterations to the above-described embodiments are possible within the scope of the present invention without departing from the scope of the present invention.

Claims (17)

1. a compound as shown in formula I or a pharmaceutically acceptable form thereof, the pharmaceutically acceptable form is a pharmaceutically acceptable salt, or a stereoisomer, or a tautomer, or hydrate, or solvate, or metabolite, or prodrug;

in, X ¹ , X ² or X ³ are independently selected from CR ⁵ or N, and at least one of X ¹ , X ² or X ³ is N; X ⁴ or X ⁵ is independently selected from C or N; R ¹ , R ² , R ³ and R ⁴ are independently selected from H, halogen, hydroxy, amino, cyano, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₁ -C ₈ aldehyde, C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₈ alkoxy or C ₁ -C ₃ Haloalkoxy, the C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₈ alkoxy Or C ₁ -C ₃ haloalkoxy is optionally substituted by one or more R ⁵ , when substituted by multiple R ⁵ , R ⁵ may be the same or different; R ⁵ is selected from H, halogen, hydroxyl, amino, cyano, C ₁ -C ₃ alkyl, C ₁ -C ₃ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl , C ₁ -C ₃ alkoxy or C ₁ -C ₃ haloalkoxy; n is selected from 0, 1, 2, 3, 4 and 5; m is selected from 0, 1, 2, 3, 4 and 5; Ring A is independently selected from C ₃ -C ₈ cycloalkyl, C ₆ -C ₁₂ spirocycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl or C ₅ -C ₁₂ heteroaryl base; Ring B is independently selected from C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl or C ₅ -C ₁₂ heteroaryl; The heteroatom or heteroatom group contained in the heteroalkyl group, heterocycloalkyl group and heteroaryl group is 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 said heteroatoms or heteroatoms being independently selected from 1, 2 and 3, respectively. 2. The compound of claim 1, or a pharmaceutically acceptable form thereof, wherein when ^X4 is N, the ring A is independently selected from the group consisting of a pyrrole ring, a cycloalkylspiropyrrole ring, a piperine pyridine ring, cycloalkylspiropiperidine ring or dihydropyrazole ring. 3. The compound of claim 2, or a pharmaceutically acceptable form thereof, wherein when X is N, the ring ^A is independently selected from

4. The compound of claim 1, or a pharmaceutically acceptable form thereof, wherein when X is C, the ring ^A is independently selected from a benzene ring, a pyridine ring, a pyrimidine ring, a pyrazine ring or pyridazine ring. 5. The compound of claim ⁴ , or a pharmaceutically acceptable form thereof, wherein when X is C, the ring A is independently selected from

6. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable form thereof, wherein when X ⁵ is N, the ring B is independently selected from pyrrole ring, piperidine ring, piperazine ring or morpholine ring. 7. The compound of claim ⁶ , or a pharmaceutically acceptable form thereof, wherein when X is N, the ring B is independently selected from

8. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable form thereof, wherein when X ⁵ is C, the ring B is independently selected from a benzene ring, a pyridine ring , pyrimidine ring, pyrazine ring or pyridazine ring. 9. The compound of claim 8, or a pharmaceutically acceptable form thereof, wherein when X ⁵ is C, the ring B is independently selected from

10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable form thereof, wherein R ¹ , R ² , R ³ and R ⁴ are each independently selected from H, Halogen, hydroxy, amino, cyano, C ₂ -C ₄ alkenyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ heteroalkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycle Alkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₃ haloalkoxy, said C ₁ -C ₄ alkyl, C ₁ -C ₄ heteroalkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₃ haloalkoxy is optionally substituted by one or more R ⁵ , when substituted by multiple R ⁵ , R ⁵ can be same or different; R ⁵ is selected from H, F, Cl, Br, hydroxy, amino, cyano, methyl, ethyl, cyclopropyl, methoxy, ethoxy, trifluoromethoxy. 11. The compound of claim 10, or a pharmaceutically acceptable form thereof, wherein R ¹ , R ² , R ³ and R ⁴ are each independently selected from H, halogen, hydroxyl, amino, cyano, methyl, ethyl, isopropyl, cyclopropyl, vinyl, methoxy, ethoxy, isopropoxy, trifluoromethyl, difluoromethyl, trifluoromethoxy, - _OCH2CHF2 , -N( _{CH3 ) 2} , -NH( _{CH3 )} or _-OCH2CF3 . 12. The compound of claim 1, or a pharmaceutically acceptable form thereof, wherein the compound of formula I is selected from the group consisting of the following formula (II), formula (III) or formula (IV) Compound:

wherein 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. 13. The compound of claim 12, or a pharmaceutically acceptable form thereof, wherein the compound represented by formula I is selected from the group consisting of compounds represented by the following formulae (II-1) to (IV-3) :

wherein 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. 14. The compound of claim 13, or a pharmaceutically acceptable form thereof, wherein the compound of formula I is any of the following compounds:

15. A pharmaceutical composition, characterized in that the pharmaceutical composition contains an effective dose of a compound according to any one of claims 1 to 14, or a pharmaceutically acceptable form thereof, and at least one drug With excipients, the pharmaceutically acceptable form is a pharmaceutically acceptable salt, or stereoisomer, or tautomer, or hydrate, or solvate, or metabolite, or prodrug. 16. A compound according to any one of claims 1 to 14, or a pharmaceutically acceptable form thereof, or the use of the pharmaceutical composition according to claim 15 in the preparation of a SHP-2 inhibitor medicine, The pharmaceutically acceptable forms are pharmaceutically acceptable salts, or stereoisomers, or tautomers, or hydrates, or solvates, or metabolites, or prodrugs. 17. A compound according to any one of claims 1 to 14, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition according to claim 15 in the manufacture of a medicament for the treatment and/or prevention of cancer the use in, the pharmaceutically acceptable form is a pharmaceutically acceptable salt, or a stereoisomer, or a tautomer, or a hydrate, or a solvate, or a metabolite, or a prodrug, so These cancers include 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, preferably non-small cell lung cancer, esophageal cancer and head and neck tumors.