WO2024199430A1 - PI3Kα INHIBITOR COMPOUND, PHARMACEUTICAL COMPOSITION, AND USE THEREOF - Google Patents

Abstract

The present invention provides a PI3Kα inhibitor compound as shown in formula I, a pharmaceutical composition, and a use thereof. The compound has good PI3Kα inhibitory activity, can be used for treating diseases related to PI3Kα, has good biological activity and good safety, and improves transmembrane activity and drug bioavailability.

Description

PI3Kα inhibitor compounds, pharmaceutical compositions and applications thereof

The present invention claims:

Priority to the prior application filed with the State Intellectual Property Office of China on March 31, 2023, with patent application number 202310344084.X and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on May 6, 2023, with patent application number 202310503871.4 and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on June 25, 2023, with patent application number 202310751715.X and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on July 21, 2023, with patent application number 202310903126.9 and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on August 23, 2023, with patent application number 202311067799.1 and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on September 27, 2023, with patent application number 202311262154.3 and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application, patent application number 202311414985.8, filed with the State Intellectual Property Office of China on October 27, 2023, entitled “PI3Kα inhibitor compounds, pharmaceutical compositions and their applications”;

Priority to the prior application, patent application number 202311605325.8, filed with the State Intellectual Property Office of China on November 28, 2023, entitled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on December 4, 2023, with patent application number 202311649417.6 and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on December 21, 2023, with patent application number 202311779783.3 and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

Priority to the prior application filed with the State Intellectual Property Office of China on February 5, 2024, with patent application number 202410166443.1 and titled “PI3Kα inhibitor compounds, pharmaceutical compositions and their uses”;

The entire content of said prior application is incorporated herein by reference.

Technical Field

The present invention belongs to the field of pharmaceutical compounds, and in particular relates to PI3Kα inhibitor compounds, pharmaceutical compositions and applications thereof.

Background Art

Phosphatidylinositol 3-kinase (PI3K) is a unique and conserved family of intracellular lipid kinases that possess both phosphatidylinositol kinase activity and serine/threonine (Ser/Thr) kinase activity. The PI3K family includes 15 kinases, which can be divided into three major categories (class I, class II, and class III) based on their structure and substrate specificity.

The most widely studied of these is class I PI3K, which is a heterodimer consisting of a regulatory subunit p85 and a catalytic subunit p110. There are four types of catalytic subunits: α, β, δ, and γ. Among them, α, β, and δ correspond to p85α, p85β, or p55 regulatory subunits; while the γ class corresponds to p101 and p84/87 regulatory subunits. The regulatory subunit has an SH2 domain that can recognize the intracellular kinase domain of RTKs and trigger the activation of the catalytic subunit p110.

Usually, after class I PI3K is activated by tyrosine kinase or G protein-coupled receptor, it can catalyze PIP2 to generate PIP3 and trigger the activation of serine/threonine kinase AKT. AKT is regulated by PDK and mTOR2, and activation of AKT can promote cell cycle progression; in addition, activation of AKT can trigger the expression of a series of downstream molecules, thereby maintaining cell survival, promoting vascular survival, and promoting cell growth. Homologous phosphatase-tensin (PTEN), as a negative regulator of PI3K signal transduction, can dephosphorylate PIP3 and convert it into PIP2.

PI3K signaling is one of the most common abnormally activated pathways in cancer and is thought to be associated with a range of human cancers. Early studies have shown that pan-PI3K inhibitors LY294002 and Wortmannin can improve cancer cells' resistance to a variety of therapies, including chemotherapy, radiotherapy, and targeted therapy. In addition to targeting cancer cells, studies have also demonstrated the potential of PI3K inhibitors in cancer immunotherapy.

In the past two decades, the development of drugs related to the PI3K pathway has been a key research area, and some results have been achieved. Several PI3K inhibitors targeting individual isoforms have now been approved by regulatory authorities. These include inhibitors targeting leukocyte-enriched PI3Kδ, which is mainly found in B-cell malignancies, and alpelisib, a PI3Kα isoform selective inhibitor for the treatment of HR+/HER2-/PIK3CA-mutated advanced metastatic breast cancer and the treatment of pro-overgrowth syndrome (PROS).

Unlike PI3Kδ and PI3Kγ, which are mainly expressed in hematopoietic cells, PI3Kα is expressed in most tissues. PI3Kα plays a core role in regulating the body's glucose homeostasis, and PI3Kα inhibition in patients usually causes hyperglycemia or hyperinsulinemia. Studies have also shown that high levels of insulin may have mitogenic and anti-apoptotic effects on cancer cells, thereby offsetting the anti-proliferative effects of PI3Kα inhibitors. Therefore, the development of PI3Kα inhibitors faces challenges in tolerability and safety.

Alpelisib has an equivalent inhibitory effect on mutant and wild-type PI3Kα. Although the drug is classified as a PI3Kα-specific drug, severe concentration-dependent side effects and drug resistance are often observed. Clinical studies have also shown that the incidence of adverse events (AEs) of Alpelisib ≥ grade 3 (mainly hyperglycemia) is high, which limits patients' tolerance and acceptance of the drug. At the same time, the sensitivity of Alpelisib depends on the PIK3CA mutation. Therefore, there is an urgent need for more accurate, more efficient, and better tolerated inhibitors for mutant PI3Kα in clinical practice to change the current treatment status and meet clinical needs.

PIK3CA is the gene encoding the PI3Kα catalytic subunit p110α protein and is the most commonly mutated gene in solid tumors. The most common hotspot mutations of the PIK3CA gene occur mainly in the kinase domain of exon 20 (H1047R) and the helical domain of exon 9 (E545K, E542K). These mutations have a greater impact on PI3Kα activity and have been shown to be oncogenic gain-of-function mutations. Among them, approximately 15% of breast cancers will have H1047R mutations, which are relatively uncommon in other tumors. Compared with PIK3CA wild-type patients, patients with PIK3CA-mutated HR+/HER2- advanced breast cancer have a poorer prognosis, poor response to traditional treatment, and resistance to endocrine therapy and chemotherapy. Some small studies that mainly analyzed ER+ breast and adenocarcinoma tumors have shown that patients with H1047R show a reduced survival rate compared with tumors containing E545K.

Therefore, targeting PI3Kα mutations and developing inhibitors with enhanced selectivity for mutant PI3Kα may provide a valuable treatment opportunity for breast cancer patients carrying this mutation, overcoming the problem of compensatory production of insulin or glucose after systemic PI3Kα inhibition. This will create an increased window for drug dosing and selectively inhibit the pathological signaling of mutant PI3Kα in cancer cells. It is hoped that the research scope of PI3Kα inhibitors will be expanded from HR+/HER2- to HER2+ and TNBC, and from late stage to early stage, so that more patients can benefit.

Summary of the invention

The present invention provides a compound represented by formula I and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt;

wherein R ₁ and R ₂ are the same or different and are independently selected from H, deuterium, the following groups which are unsubstituted or optionally substituted by one, two or more R ₁₁ : C _1-12 alkyl, C _1-12 alkyloxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkyloxy, C _3-12 cycloalkyl; each R ₁₁ is the same or different and is independently selected from H, deuterium, halogen, CN, OH, C _1-12 alkyl;

each R ₃ is the same or different and is independently selected from H, deuterium, halogen, CN, OH, the following groups which are unsubstituted or optionally substituted with one, two or more R ₃₁ : C _1-12 alkyl, C _1-12 alkyloxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkyloxy, C _3-12 cycloalkyl, 3-14 membered heterocyclyl, C _6-14 aryl, 5-14 membered heteroaryl, N(R ₃₂ )(R ₃₃ ); each R ₃₁ is the same or different and is independently selected from H, deuterium, halogen, CN, C _1-12 alkyl, C _1-12 alkyloxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkyloxy, C _1-12 acyl; R ₃₂ and R ₃₃ are the same or different and are independently selected from H, deuterium, C _1-12 alkyl, S(＝O) ₂ R _311, S(=O)(=NH)R ₃₁₂ ; R _{311 and} R ₃₁₂ are the same or different and are independently selected from H, deuterium, and C _1-12 alkyl;

X is selected from N, NR _x1 or CR _x2 ; R _x1 and R _x2 are the same or different and are independently selected from H, deuterium, halogen, CN, C _1-12 alkyl;

Y is selected from O, S or N;

Ring A is selected from a C _3-12 carbocyclic ring, a 3-14 membered heterocyclic ring, a C _6-14 aromatic ring, and a 5-14 membered heteroaromatic ring;

Each _Ra is the same or different and is independently selected from H, deuterium, halogen, CN, OH, _C1-12 alkyl, _C1-12 alkyloxy, _C3-12 cycloalkyl, halogenated _C1-12 alkyl, halogenated _C1-12 alkyloxy, OH- _C1-12 alkyl;

E is absent or selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : _C3-12 cycloalkyl, 3-14 membered heterocyclyl, N( _Re1 )( _Re2 ), _C1-12 alkyl-CN; each _Re is the same or different and is independently selected from H, deuterium, halogen, CN, _C1-12 alkyl-CN, OH, _C1-12 alkyl, _C1-12 alkyloxy, _C3-12 cycloalkyl, halogenated _C1-12 alkyl, halogenated _C1-12 alkyloxy, halogenated _C3-12 cycloalkyl; _Re1 and _Re2 are the same or different and are independently selected from H, deuterium, _C1-12 alkyl, _C1-12 alkyl- _C3-12 cycloalkyl, _C3-12 cycloalkyl- _C1-12 alkyl;

m is selected from 0, 1, 2, 3 or 4;

n is selected from 0, 1, 2, 3 or 4;

p is selected from 0, 1, 2, 3 or 4.

According to some embodiments, R ₁ and R ₂ are the same or different and are independently selected from H, unsubstituted or the following groups optionally substituted by one, two or more R ₁₁ : C _1-6 alkyl, halogenated C _1-6 alkyl, C _3-6 cycloalkyl;

According to some embodiments, each R ₁₁ is the same or different and is independently selected from H, halogen, C _1-6 alkyl;

According to some embodiments, each R ₁₁ is the same or different and is independently selected from H, F, methyl;

According to some embodiments, R ₁ and R ₂ are the same or different and are independently selected from H, methyl, ethyl, isopropyl, trifluoromethyl, cyclopropyl, tert-butyl, methylcyclopropyl (such as 1-methyl-cyclopropyl-1-yl), fluorocyclopropyl (such as 1-fluoro-cyclopropyl-1-yl).

According to some embodiments, each R ₃ is the same or different and is independently selected from H, halogen, CN, unsubstituted or the following groups optionally substituted with one, two or more R ₃₁ : C _1-6 alkyl, C _1-6 alkyloxy, halo C _1-6 alkyl, halo C _1-6 alkyloxy, C _3-6 cycloalkyl, 3-8 membered heterocyclyl, C _6-10 aryl, 5-10 membered heteroaryl, N(R ₃₂ )(R ₃₃ );

According to some embodiments, each R ₃₁ is the same or different and is independently selected from H, CN, C _1-6 alkyl, C _1-6 acyl;

According to some embodiments, each R ₃₁ is the same or different and is independently selected from H, C _1-6 alkyl; for example methyl;

According to some embodiments, R ₃₂ and R ₃₃ are the same or different and are independently selected from H, C _1-6 alkyl, -S(=O) ₂ -C _1-6 alkyl, -S(=O)(=NH)C _1-6 alkyl;

According to some embodiments, R ₃₂ and R ₃₃ are the same or different and are independently selected from H, methyl, S(═O) ₂ CH ₃ , S(═O)(═NH)CH ₃ .

According to some embodiments, each R ₃ is the same or different and is independently selected from H, F, Cl, CN, methyl, methoxy, difluoromethoxy, trifluoromethoxy, methylamino, -NH-S(=O) ₂ CH ₃ , -NH-S(=O)(=NH)CH ₃ , morpholinyl (e.g. ), tetrahydropyrrolyl (such as ), phenyl, methylpyrazolyl (such as ), cyclopropyl;

According to some embodiments, X is selected from N, NR _x1 or CR _x2 ; R _x1 and R _x2 are the same or different and are independently selected from H, halogen, CN, C _1-6 alkyl;

According to some embodiments, X is selected from N, NR _x1 or CR _x2 ; R _x1 and R _x2 are the same or different and are independently selected from H, F, Cl, CN, methyl, ethyl.

According to some embodiments, Ring A is selected from a C _3-8 carbocyclic ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring, a 5-10 membered heteroaromatic ring;

According to some embodiments, ring A is selected from a pyrimidine ring, a benzene ring, a pyridine ring, a pyrazine ring, a pyridazine ring, a thiophene ring, a furan ring, a pyrazole ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, a triazole ring, a quinoline ring, a quinazoline ring, a pyrrolopyridine ring (such as ), tetrahydroquinoline ring (such as ), cyclopentadienylpyridine ring (such as ), pyrazolopyrimidine ring (such as ), naphthyridine ring (such as ), pyrazolopyridine ring (such as ), Imidazolopyridine ring (such as ), quinoline ring (such as ), indazole ring (such as ), benzimidazole ring (such as ), triazolopyridine ring (such as ).

According to some embodiments, each _Ra is the same or different and is independently selected from H, deuterium, halogen, CN, OH, _C1-6 alkyl, _C1-6 alkyloxy, _C3-6 cycloalkyl, halo- _C1-6 alkyl, halo- _C1-6 alkyloxy, OH- _C1-6 alkyl;

According to some embodiments, each _Ra is the same or different and is independently selected from H, deuterium, halogen, CN, OH, _C1-3 alkyl, _C1-3 alkyloxy, _C3-6 cycloalkyl, halogenated _C1-3 alkyl, halogenated _C1-3 alkyloxy, OH- _C1-3 alkyl;

According to some embodiments, each _Ra is the same or different and is independently selected from H, _C1-6 alkyl, halo- _C1-6 alkyl, OH- _C1-6 alkyl; for example H, methyl, 2-hydroxyethyl.

According to some embodiments, E is absent and p is selected from 0;

According to some embodiments, E is selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, N( _Re1 )( _Re2 ), _C1-6 alkyl-CN,;

According to some embodiments, E is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Re : 3-10 membered heterocyclyl, N( _Re1 )( _Re2 );

According to some embodiments, E is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Re : 3-8 membered heterocyclyl, N( _Re1 )( _Re2 );

According to some embodiments, E is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Re : _NH2 , azetidinyl, piperidinyl, tetrahydropyrrolyl, morpholinyl, _{CH2CH2CN ,}

According to some embodiments, E is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Re : _NH2 , azetidinyl, piperidinyl, tetrahydropyrrolyl, morpholinyl,

According to some embodiments, each _Re is the same or different and is independently selected from H, halogen, OH, _C1-6 alkyl-CN, _C1-6 alkyl, _C1-6 alkyloxy, halogenated _C1-6 alkyl, halogenated _C1-6 alkyloxy, _C3-6 cycloalkyl;

According to some embodiments, each _Re is the same or different and is independently selected from H, F, OH, _CH2- CN, methyl, ethyl, n-propyl, isopropyl, tert-butyl, methoxy, difluoromethoxy, cyclopropyl;

According to some embodiments, each _Re is the same or different and is independently selected from H, halogen, OH, _C1-6 alkyl, _C1-6 alkyloxy, halogenated _C1-6 alkyl, halogenated _C1-6 alkyloxy, _C3-6 cycloalkyl;

According to some embodiments, each _Re is the same or different and is independently selected from H, F, OH, methyl, ethyl, n-propyl, isopropyl, tert-butyl, methoxy, difluoromethoxy, and cyclopropyl.

According to some embodiments, _Re1 and _Re2 are the same or different and are independently selected from H, C _1-6 alkyl, C _1-6 alkyl-C _3-8 cycloalkyl, C _3-8 cycloalkyl-C _1-6 alkyl.

According to some embodiments, the compound represented by Formula I may have the structure shown below:

wherein R ₁ , R ₂ , R ₃ , X, Y, Ring A, E, _Ra , _Re , m, n, and p have the meanings described herein.

According to some embodiments, the compound represented by Formula I may have the structure shown below:

wherein R ₁ , R ₂ , R ₃ , X, Y, Ring A, E, _Ra , _Re , m, n, and p have the meanings described herein.

According to some embodiments, the compound represented by Formula I may have the structure shown below:

wherein E, _Re , _R3 , _Ra , n, and p have the definitions given herein.

According to some embodiments, the compound represented by Formula I may have the structure shown below:

Wherein, _Re and p have the definitions described herein.

According to some embodiments, the compound represented by Formula I may have the structure shown below:

The present invention also provides a method for preparing the compound represented by formula I, comprising the following steps:

wherein R ₁ , R ₂ , R ₃ , _Ra , _Re , ring A, E, X, Y, m, n, p have the definitions described herein; Z is selected from a leaving group such as halogen; and R is selected from C _6-10 aryl, C _6-10 aryl-C _1-3 alkyl, C _1-3 alkyl-C _6-10 aryl, such as phenyl, tolyl, benzyl.

The present invention further provides a pharmaceutical composition comprising the compound of formula I described in the present invention and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof.

In some embodiments, the pharmaceutical composition described in the present invention further comprises a therapeutically effective amount of the compound of formula I described in the present invention and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

The carrier in the pharmaceutical composition is "acceptable" in that it is compatible with the active ingredient of the composition (and preferably, capable of stabilizing the active ingredient) and not deleterious to the subject being treated. One or more pharmaceutical excipients may be used for delivery of the active compound.

The present invention further provides the use of the compound of formula I and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of PI3Kα inhibitors.

The present invention further provides the use of the compound of formula I and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a drug for preventing and/or treating cancer, such as lung cancer, gastric cancer, endometrial cancer, ovarian cancer, bladder cancer, breast cancer, colon cancer, brain cancer, prostate cancer, skin cancer and/or benign overgrowth syndrome.

The present invention further provides the use of the compound of formula I and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a drug for preventing and/or treating PIK3CA-associated overgrowth (PROS).

The present invention also provides a method for preventing and/or treating a PI3Kα-mediated disease or condition, which comprises administering to a patient in need of such treatment a therapeutically effective amount of at least one compound or pharmaceutical composition of the present invention alone, or, optionally, in combination with another compound of the present invention and/or at least one other type of therapeutic agent.

According to some embodiments, the PI3Kα-mediated disease or condition is selected from cancer, such as lung cancer, gastric cancer, endometrial cancer, cancer, ovarian cancer, bladder cancer, breast cancer, colon cancer, brain cancer, prostate cancer, skin cancer, and/or benign overgrowth syndrome;

According to some embodiments, the PI3Kα-mediated disease or condition is PIK3CA-associated overgrowth (PROS).

Beneficial Effects

The compounds provided by the present invention have good PI3Ka inhibitory activity and can be used to treat diseases related to PI3Kα. The compounds of the present invention not only have good biological activity, but also improve the in vivo pharmacokinetic properties of such compounds, enhance the drugability of the compounds, and have good in vivo efficacy and good safety.

Definition and explanation of terms

Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and combined with each other. The definitions of groups and compound structures after such combinations and combinations should be understood to be within the scope of the specification and/or claims of this application.

The term "optional" (or "optionally", "optionally") in the general formula definitions of the present application means the situation of being substituted by zero, one or more substituents, for example, "optionally substituted by one, two or more R" means that it may not be substituted by R (unsubstituted) or may be selectively substituted by one, two or more R.

"More" means three or more.

Unless otherwise specified, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-12" is equivalent to describing each integer value in the numerical range "1-12", i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

The term "C _1-12 alkyl" is understood to mean straight-chain and branched alkyl groups having 1 to 12 carbon atoms, "C _1-8 alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and "C _1-6 alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl or the like or isomers thereof.

The term "C _3-12 cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (such as condensed, bridged, spiro) hydrocarbon ring or tricyclic alkane having 3 to 12 carbon atoms, preferably a "C _3-10 cycloalkyl", more preferably a "C _3-8 cycloalkyl". The term "C _3-12 cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (such as bridged, spiro) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The _C3-12 cycloalkyl group may be a monocyclic hydrocarbon group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group, such as borneol, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonanyl, 2,6-diazaspiro[3,4]octanyl, or a tricyclic hydrocarbon group, such as adamantyl.

The term "C _6-14 aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic ring having 6 to 14 carbon atoms, A bicyclic (eg, fused, bridged, spiro) or tricyclic hydrocarbon ring, which may be a single aromatic ring or multiple aromatic rings fused together, is preferably a "C _6-10 aryl group". The term "C _6-14 aryl" is to be understood as preferably meaning a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C _6-14 aryl") having 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms, in particular a ring having 6 carbon atoms ("C ₆ aryl"), such as phenyl; or biphenyl, or a ring having 9 carbon atoms ("C ₉ aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C ₁₃ aryl"), such as fluorenyl, or a ring having 14 carbon atoms ("C ₁₄ aryl"), such as anthracenyl. When the C _6-20 aryl is substituted, it may be mono- or polysubstituted. Furthermore, there is no limitation on the substitution site, and for example, substitution may be at the ortho, para or meta position.

The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic (e.g. fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 14 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, for example "5-10 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3 heteroatoms each independently selected from N, O and S and, in each case, furthermore, may be benzo-fused. "Heteroaryl" also refers to a radical in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3- , 4-, 5-, 6-, 7- or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolylcarbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-phenanthridinyl, -acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-oxadiazole, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthroline, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl , 2-, 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzoisoquinolyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]pyranyl , 2-, 3-, 4-, 5-, 7-, or 8-5H-pyrido[2,3-d]-oxazinyl, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or 54H-imidazo[4,5-d]thiazolyl, 3-, 5-, or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5-, or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8-, or 9-imidazo[2,1-b]thiazolyl, -furano[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl, 2-, 4- , 4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-4H-pyrrolo[1,2-b][2]benzazepinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When other groups are connected to form the compounds of the present invention, the carbon atoms on the 5-14 membered heteroaryl ring may be connected to other groups, or the heteroatoms on the 5-14 membered heteroaryl ring may be connected to other groups. When the 5-14 membered heteroaryl is substituted, it may be monosubstituted or polysubstituted. In addition, there is no limitation on the substitution site, for example, the hydrogen connected to the carbon atom on the heteroaryl ring may be substituted, or the hydrogen connected to the heteroatom on the heteroaryl ring may be substituted.

Unless otherwise defined, the term "3-14 membered heterocyclyl" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, 6- or 7-membered monocyclic ring, a 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring (such as a fused ring, a bridged ring, a spirocyclic ring) or a 10-, 11-, 12-, 13- or 14-membered tricyclic ring system, and contains at least one, for example 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S may also be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)- or -S(O) ₂ -. For example, the "3-14 membered heterocyclyl" may be a 3-14 membered N-containing heterocyclyl (containing at least one N). Preferably, the heterocyclyl may be selected from a "3-10 membered heterocyclyl". The term "3-10 membered heterocyclyl" means a saturated or unsaturated non-aromatic ring or ring system, and contains at least one heteroatom selected from O, S and N. The heterocyclyl may be connected to the rest of the molecule through any one of the carbon atoms or the nitrogen atom (if present). The heterocyclyl may include fused or bridged rings and spirocyclic rings. In particular, the heterocyclyl may include, but is not limited to: a 4-membered ring, such as azetidinyl, oxetanyl; a 5-membered ring, such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a 7-membered ring, such as diazepanyl. Optionally, the heterocyclyl may be benzo-fused. The heterocyclic group may be bicyclic, for example but not limited to a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl ring. The heterocyclic group may be partially unsaturated, i.e., it may contain one or more double bonds, for example but not limited to dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, for example but not limited to dihydroisoquinolinyl. When the 3-14-membered heterocyclic group is connected to other groups to form the compound of the present invention, the carbon atoms on the 3-14-membered heterocyclic group may be connected to other groups, or the heterocyclic atoms on the 3-14-membered heterocyclic group may be connected to other groups. For example, when the 3-14 membered heterocyclic group is selected from piperazinyl, the nitrogen atom on the piperazinyl group may be connected to other groups. Or when the 3-14 membered heterocyclic group is selected from piperidinyl group, the nitrogen atom on the piperidinyl ring and the carbon atom at the para position thereof may be connected to other groups.

The term "spirocyclic" refers to a ring system in which two rings share one ring-forming atom.

The term "fused ring" refers to a ring system in which two rings share two ring atoms.

The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

The term "halogen" refers to fluorine, chlorine, bromine and iodine.

"Halo" means substituted with one or more halogens.

The term "oxo" refers to a substituent in which a carbon atom, a nitrogen atom or a sulfur atom is oxidized to form an oxy group (=O).

The term "alkylamino" refers to -NH-(alkyl) or -N-(alkyl)2, wherein alkyl is as defined above. Non-limiting examples of alkylamino include methylamino, ethylamino, propylamino, isopropylamino, butylamino, dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, and the like.

The term "alkyloxy" refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy. Alkoxy may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkyloxy, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy or heterocycloalkyloxy.

The terms "alkyleneoxy" and "oxyalkylene" refer to -alkylene-O- or -O-alkylene-, wherein alkylene represents a straight or branched saturated divalent hydrocarbon group. The definition of the number of carbon atoms of "alkylene" applies to the definition of "alkyl" above. It will be understood by those skilled in the art that alkyleneoxy or oxyalkylene can be attached to the rest of the molecule containing it in any direction, that is, the two can be used interchangeably.

The term "nitrogen oxide" refers to a compound formed by oxidation of a nitrogen atom in a tertiary amine or nitrogen-containing (aromatic) heterocyclic compound structure.

Unless otherwise specified, heterocyclic groups, heteroaryls or heteroarylene groups include all possible isomeric forms thereof, such as positional isomers thereof. Thus, for some illustrative non-limiting examples, forms substituted or bonded to other groups at 1, 2 or more positions in the 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, etc. (if present) may include pyridin-2-yl, pyridin-2-ylene, pyridin-3-yl, pyridin-3-ylene, pyridin-4-ylene and pyridin-4-ylene; thienyl or thienylene groups include thien-2-yl, thien-2-ylene, thien-3-ylene and thien-3-ylene; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl.

Wavy lines intersecting chemical bonds Used to indicate the connection position between a group and other atoms in a molecular structure. Indicates connection to the 3-position of the pyridyl group. When the group connection position is not fixed, taking the pyridyl group as an example, The above-mentioned method shows that it can be connected to any connectable position on the pyridyl group. Unless otherwise specified, similar expressions in this application are interpreted the same as above.

In the chemical structure of the compound described in the present invention, the bond Indicates that the configuration is not specified. or Indicates the absolute configuration, that is, if there are stereoisomers in the chemical structure, the bond Can be or or include both and Two configurations.

In the present invention, the compounds involved also include isotopically labeled compounds, which are the same as those shown in Formula I, but one or more atoms are replaced by atoms having atomic masses or mass numbers different from the atomic masses or mass numbers usually occurring in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F and Cl, such as ² H, ³ H, ¹³ C, 11 C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl, respectively. Compounds of the present invention, prodrugs thereof, or pharmaceutically acceptable salts of the compounds or prodrugs containing the above-mentioned isotopes and/or other isotopes of other atoms are within the scope of ^the present invention. Certain isotopically labeled compounds of the present invention, for example compounds incorporating radioactive isotopes (such as ³ H and ¹⁴ C), can be used for drug and/or substrate tissue distribution assays. Tritiated (i.e., ³ H) and carbon 14 (i.e., ¹⁴ C) isotopes are particularly preferred for ease of preparation and detectability. Furthermore, substitution with heavier isotopes (such as deuterium, i.e., ² H or D) may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and therefore may be preferred in some circumstances. The presence of hydrogen in a substituent of the present invention without the term deuterium or tritium being separately listed does not exclude deuterium or tritium, but may also include deuterium or tritium.

Those skilled in the art will appreciate that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form To form base addition salts; if these compounds contain both an acidic center (for example a carboxyl group) and a basic center (for example an amino group), they can also form internal salts.

The compounds of the present invention may exist in the form of solvates (e.g., hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of the polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

According to its molecular structure, the compounds of the present invention may be chiral, and therefore various enantiomeric forms may exist. Thus, these compounds may exist in racemic form or optically active form. The compounds of the present invention encompass isomers or mixtures, racemates in which each chiral carbon is in R or S configuration. The compounds of the present invention or their intermediates can be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolution agent. Examples of suitable resolution agents are optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids in R and S forms. Chromatographic enantiomer resolution can also be advantageously carried out with the aid of optically active resolving agents such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers immobilized on silica gel. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example, hexane/isopropanol/acetonitrile.

The corresponding stable isomers can be separated according to known methods, for example by extraction, filtration or column chromatography.

The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

The term "therapeutically effective amount" refers to the amount of an active compound or drug that elicits the biological or medical response that a researcher, veterinarian, physician or other clinician is seeking in a tissue, system, animal, individual or human, and includes one or more of the following: (1) Preventing disease: e.g., preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but does not yet experience or develop the pathology or symptoms of the disease. (2) Inhibiting disease: e.g., inhibiting a disease, disorder or condition (i.e., preventing further development of the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition. (3) Alleviating disease: e.g., alleviating a disease, disorder or condition (i.e., reversing the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1: Efficacy of the test compounds in the xxT47D subcutaneous tumor model established in Balb/c nude mice.

Figure 2: Changes in insulin levels after administration of the test compounds.

DETAILED DESCRIPTION

The technical solution of the present disclosure will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary illustrations and explanations of the present disclosure and should not be construed as limiting the scope of protection of the present disclosure. All technologies implemented based on the above content of the present disclosure are included in the scope of protection intended by the present disclosure.

Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

Example 1

3-[1-(5,7-difluoro-3-methyl-1-benzothiophen-2-yl)-2,2,2-trifluoroethyl]-1-[2-(3-fluoroazetidin-1-yl)pyrimidin-5-yl]urea 001

The first step 1-(3,5-difluoro-2-mercaptophenyl)ethanone 001b

Under nitrogen protection, aluminum chloride (4 g) was added to a toluene (50 mL) solution of 001a (7 g, prepared with reference to "WO2023288242A1") at 0°C. After the addition was complete, it was stirred at 0°C for half an hour. The reaction mixture was diluted with ethyl acetate (100 mL) and extracted with sodium hydroxide solution (2M, 2×100 mL). The aqueous phase was acidified with dilute hydrochloric acid (6N, 100 mL), extracted with ethyl acetate (3×100 mL), and the organic phase was dried over anhydrous sodium sulfate. After filtration, the residue was concentrated under reduced pressure to give compound 001b (4 g).

LC-MS (ESI, m/z)=186.90[MH] ^-

Step 2 5,7-Difluoro-3-methyl-1-benzothiophene-2-carboxylic acid ethyl ester 001c

To a solution of 001b (4 g) and potassium carbonate (5.9 g) in acetonitrile (60 mL) was added dropwise ethyl bromoacetate (3.6 g) at 0°C. After the addition was complete, the mixture was stirred at 80°C overnight. The resulting mixture was filtered, the filter cake was washed with ethyl acetate (2×20 mL), the organic phase was dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-17%) to obtain compound 001c (4 g).

¹ H NMR (400MHz, DMSO-d ₆ ) δ7.83-7.76 (m, 1H), 7.64-7.55 (m, 1H), 4.36 (q, 2H), 2.71 (s, 3H), 1.34 (t, 3H) ).

Step 3 5,7-difluoro-3-methyl-1-benzothiophene-2-methanol 001d

Under nitrogen protection, lithium aluminum tetrahydride (0.5 g) was added to tetrahydrofuran (30 mL) at 0°C, and then 001c (3.5 g) was added dropwise. After the addition was complete, stirring was continued at room temperature for 1 hour. The reaction mixture was quenched by adding sodium sulfate decahydrate (5 g) at 0°C. The mixture was filtered, the filter cake was washed with ethyl acetate (30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 001d (2.8 g).

¹ H NMR (400MHz, DMSO-d ₆ ) δ7.49-7.45 (m, 1H), 7.32-7.25 (m, 1H), 5.77 (t, 1H), 4.76 (t, 2H), 2.28 (s, 3H ).

Step 4: 5,7-difluoro-3-methyl-1-benzothiophene-2-carbaldehyde 001e

To a solution of 001d (2.8 g) in dichloromethane (20 mL) at 0°C, Dess-Martin periodinane (5.5 g) was added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate solution (50 mL). Extracted with dichloromethane (3×50 mL). The organic phases were combined and backwashed with saturated aqueous sodium chloride solution (50 mL), and the organic phases were dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-20%) to give compound 001e (2.3 g).

LC-MS (ESI, m/z):=213.00[M+H] ⁺

Step 5: N-[(1E)-(5,7-difluoro-3-methyl-1-benzothiophen-2-yl)methylene]-2-methylpropane-2-sulfenamide 001f

At room temperature, tetraethyl titanate (7.5 g) was added to tetrahydrofuran (40 mL) of 001e (2.3 g) and tert-butylsulfenamide (2 g). After the addition was complete, the atmosphere was replaced with nitrogen three times and stirred at 80°C overnight. The resulting mixture was quenched with a saturated aqueous sodium chloride solution (20 mL) at room temperature. The resulting mixture was filtered and the filter cake was washed with ethyl acetate (1×20 mL). The filtrate was extracted with ethyl acetate (3×40 mL). The organic phases were combined and then backwashed with a saturated sodium chloride solution (40 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-50%) to obtain compound 001f (3 g).

LC-MS (ESI, m/z):=316.00[M+H] ⁺

Step 6: N-[1-(5,7-difluoro-3-methyl-1-benzothiophene-2-yl)-2,2,2-trifluoroethyl]-2-methylpropane-2-sulfenamide 001g

Under nitrogen protection, tetrabutylammonium difluorotriphenyl silicate (5g) was added to a solution of 001f (3g) in tetrahydrofuran (30mL) at room temperature, the temperature was lowered to -60°C, and stirring was continued for 1 hour. Then (trifluoromethyl)trimethylsilane (5.5g) was added dropwise. After the addition was completed, stirring was continued at -60°C for 1 hour. Then the temperature was raised to -30°C and stirred for 1 hour. Then saturated aqueous ammonium chloride solution (50mL) was added at -30°C to quench. The reaction mixture was extracted with ethyl acetate (3×50mL). The organic phases were combined, backwashed with saturated sodium chloride solution (50mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, ethyl acetate/petroleum ether (0-19%) to obtain compound 001g (2.2g).

LC-MS (ESI, m/z):=385.95[M+H] ⁺

Step 7 1-(5,7-difluoro-3-methyl-1-benzothiophene-2-yl)-2,2,2-trifluoroethylamine 001h

At room temperature, 001g (2.2g) was added to a solution of 1,4-dioxane (4M, 20mL) in hydrogen chloride and stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and then the pH was adjusted to 8 with a saturated sodium bicarbonate aqueous solution, extracted with dichloromethane (2×50mL), and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure to obtain compound 001h (1.4g).

LC-MS (ESI, m/z):=264.95[M-16] ⁺

Step 8 3-[1-(5,7-difluoro-3-methyl-1-benzothiophene-2-yl)-2,2,2-trifluoroethyl]-1-[2-(3-fluoroazetidine-1-yl)pyrimidin-5-yl]urea 001

At 80°C, a solution of 001i (35.88 mg) and phenyl chloroformate (3 mL) in pyridine (3 mL) was stirred for 1 hour. The reaction solution was concentrated, and the resulting residue was dissolved in N, N-dimethylformamide solution (3 mL), and then 001h (50 mg) and N, N-diisopropylethylamine (68.93 mg) were added. After the addition was complete, the mixture was stirred at 60°C for 1 hour. The reaction mixture was filtered, and the filter cake was washed with N, N-dimethylformamide (1 mL). The crude product was purified by high-performance liquid chromatography (chromatographic column specifications: Kinetex EVO C18 column, 30 mm*150 mm, 5 μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; gradient: 10% B to 39% B in 8 minutes) to obtain compound 001 (13.51 mg).

LC-MS (ESI, m/z):=475.80[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.41 (s, 2H), 8.32 (s, 1H), 7.90 (d, 1H), 7.66-7.60 (m, 1H), 7.51-7.43 (m, 1H) ), 6.26-6.15(m, 1H), 5.58-5.35(m, 1H), 4.37-4.25(m, 2H), 4.08-3.96(m, 2H), 2.46(s, 3H).

Synthesis of intermediate 001i

2-(3-Fluoroazetidin-1-yl)-5-nitropyrimidine 001k

Under nitrogen protection, potassium carbonate (12.99 g) was added to a mixed solution of compound 001j (5 g) and 3-fluoroazetidine hydrochloride (4.20 g) in N, N-dimethylformamide (50 mL) at room temperature. The temperature was then raised to 90°C and the reaction was stirred for 3 hours. The reaction solution was then cooled to room temperature. The reaction mixture was diluted with water (200 mL) and then extracted with ethyl acetate (3×200 mL). The organic phases were combined and backwashed with saturated brine (1×300 mL), and the organic phases were dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, ethyl acetate/petroleum ether (0-40%) to obtain compound 001k (2.5 g).

LC-MS: (ESI, m/z)=199.35[M+H] ⁺

2-(3-Fluoroazetidin-1-yl)pyrimidin-5-amine 001i

Palladium carbon (1.15 g) was added to a solution of compound 001k (2.3 g) in methanol (25 mL) at room temperature, and then stirred in a hydrogen atmosphere for 3 hours. The mixture was filtered, the filter cake was washed with methanol (3×30 mL), and the filtrate was concentrated under reduced pressure to obtain compound 001i (1.5 g).

LC-MS: (ESI, m/z)=169.10[M+H] ⁺

Example 2

3-[1-(5,7-difluoro-3-methyl-1-benzothiophen-2-yl)-2,2,2-trifluoroethyl]-1-{2-[(2-hydroxy-2-methylpropyl)amino]pyrimidin-5-yl}urea 002

Step 1 2-Methyl-1-[(5-nitropyrimidin-2-yl)amino]propan-2-ol 002b

Under nitrogen protection, 1-amino-2-methylpropan-2-ol (670.50 mg) and potassium carbonate (2.60 g) were added to a solution of 002a (1 g) in N, N-dimethylformamide (10 mL) at room temperature, and the mixture was stirred at 80°C overnight. The reaction mixture was cooled to room temperature, diluted with water (30 mL), and extracted with ethyl acetate (3×30 mL). The organic phases were combined and backwashed with saturated brine (1×60 mL), the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-30%) to obtain compound 002b (1 g).

LC-MS: (ESI, m/z)=213.05[M+H] ⁺

Step 2 1-[(5-aminopyrimidin-2-yl)amino]-2-methylpropan-2-ol 002c

Palladium carbon (500 mg) was added to a solution of compound 002b (1 g) in methanol (10 mL). The reaction was stirred at room temperature for 3 hours under hydrogen protection. The mixture was filtered, the filter cake was washed with methanol (3×10 mL), and the filtrate was concentrated under reduced pressure to give compound 002c (800 mg). The crude product was used directly in the next step without purification.

LC-MS: (ESI, m/z)=183.25[M+H] ⁺

Step 3 3-[1-(5,7-difluoro-3-methyl-1-benzothiophene-2-yl)-2,2,2-trifluoroethyl]-1-{2-[(2-hydroxy-2-methylpropyl)amino]pyrimidin-5-yl}urea 002

To a solution of 002c (39 mg) in N, N-dimethylformamide (3 mL) was added phenyl chloroformate (33 mg) at room temperature, stirred for 1 h, and then compound 001h (50 mg) was added, and the reaction was stirred at 60°C for 2 h. The reaction mixture was filtered, and the filter cake was washed with N, N-dimethylformamide (1 mL). The crude product was purified by high-performance liquid chromatography (chromatographic column specifications: Kinetex EVO C18 column, 30 mm*150 mm, 5 μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; gradient: 40% B to 58% B in 8 minutes) to obtain compound 002 (13.20 mg).

LC-MS(ESI, m/z)=489.80[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.24 (s, 2H), 8.12 (s, 1H), 7.85 (d, 1H), 7.65-7.60 (m, 1H), 7.50-7.43 (m, 1H) ), 6.58(t, 1H), 6.24-6.13(m, 1H), 4.51(brs, 1H), 3.25(d, 2H), 2.45(s, 3H), 1.08(s, 6H).

Example 3

3-[1-(5-fluoro-3-methyl-1-benzothiophen-2-yl)-2-methylpropyl]-1-[2-(3-fluoroazetidin-1-yl)pyrimidin-5-yl]urea 003

The first step 1-(5-fluoro-3-methyl-1-benzothiophen-2-yl)-2-methyl-1-propanone 003b

1-Bromo-3-methylbutane-2-one (1.75 g) was added to a solution of 003a (1.8 g, prepared by reference to "CN115403579A") and potassium carbonate (2.92 g) in acetonitrile (40 mL). After the addition was complete, the mixture was stirred at 80°C overnight. The mixture was filtered and the filter cake was washed with ethyl acetate (1*10 mL). The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-30%) to obtain compound 003b (2 g).

LC-MS (ESI, m/z):=236.95[M+H] ⁺

Step 2 1-(5-Fluoro-3-methyl-1-benzothiophen-2-yl)-2-methylpropan-1-amine 003c

At room temperature, add ammonium acetate (1631 mg) and methanol (5 mL) solution to 003b (500 mg), and add anhydrous sodium sulfate (300 mg). After the addition is complete, stir at room temperature for 1 hour. Then add sodium cyanoborohydride (133 mg). After the addition is complete, stir at 80°C overnight. The reaction solution is concentrated under reduced pressure, and the resulting residue is purified by silica gel column chromatography, methanol/dichloromethane (0-20%) to obtain compound 003c (400 mg).

LC-MS (ESI, m/z):=221.00[M-16] ⁺

Step 3 3-[1-(5-fluoro-3-methyl-1-benzothiophene-2-yl)-2-methylpropyl]-1-[2-(3-fluoroazetidin-1-yl)pyrimidin-5-yl]urea 003

At room temperature, N, N-diisopropylethylamine (82 mg) was added to a solution of phenyl chloroformate (40 mg) and 001i (43 mg) in N, N-dimethylformamide (3 mL). After the addition was complete, the mixture was stirred for 1 hour. Then 003c (50 mg) was added. The temperature was raised to 60°C and stirring was continued for 1 hour. The mixture was filtered, the filter cake was washed with N, N-dimethylformamide (1 mL), and the filtrates were combined. The crude product was purified by high-performance liquid chromatography (chromatographic column specifications: Kinetex EVO C18 column, 30 mm*150 mm, 5 μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; gradient: 30% B to 70% B in 8 minutes) to obtain compound 003 (13.51 mg, 15.9%)

LC-MS (ESI, m/z):=431.90[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.38 (s, 2H), 8.20 (s, 1H), 7.94-7.88 (m, 1H), 7.55-7.49 (m, 1H), 7.23-7.16 (m , 1H), 6.86 (d, 1H), 5.56-5.35 (m, 1H), 4.90 (t, 1H), 4.36-4.23 (m, 2H), 4.07-3.94 (m, 2H), 2.34 (s, 3H ), 2.07-1.96 (m, 1H), 1.02 (d, 3H), 0.87 (d, 3H).

Example 4

1-(2-(azetidin-1-yl)pyrimidin-5-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 050

The first step is (2-chloropyrimidin-5-yl) phenyl carbamate 050b

At room temperature, compound 050a (1 g) and phenyl chloroformate (1.45 g) were dissolved in tetrahydrofuran (15 mL), stirred at room temperature for 1 hour, and the resulting mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-20%) to obtain compound 050b (1.2 g).

LC-MS: (ESI, m/z)=250.10[M+H] ⁺

Step 2 1-(2-chloropyrimidin-5-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 050c

Under nitrogen protection, compound 050b (466 mg) was added to a solution of compound 001h (500 mg) in pyridine (5 mL) at room temperature, and the reaction was stirred at 80°C overnight. The reaction mixture was cooled to room temperature. The obtained residue was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-20%) to obtain compound 050c (400 mg).

LC-MS: (ESI, m/z)=437.00[M+H] ⁺

Step 3 1-(2-(azetidin-1-yl)pyrimidin-5-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 050

Compound 050c (40 mg) and azetidine (11 mg) were dissolved in anhydrous ethanol (1 mL), heated to 80°C and stirred overnight. The resulting mixture was filtered and the filter cake was washed with anhydrous ethanol (2×1 mL). The resulting mixture was purified by high performance liquid chromatography (chromatographic column specifications: XBridge BEH C18 OBD Prep Column 130, 5 μm column, 30*150 mm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; gradient: 40% B to 60% B in 8 minutes) to obtain compound 050 (18.44 mg).

LC-MS: (ESI, m/z)=457.80[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.34 (s, 2H), 8.24 (s, 1H), 7.85 (d, 1H), 7.65-7.60 (m, 1H), 7.50-7.42 (m, 1H) ), 6.25-6.14(m, 1H), 4.01-3.93(m, 4H), 2.45(s, 3H), 2.32-2.22(m, 2H).

Example 5

1-(2-(5-oxa-2-azaspiro[3.4]octan-2-yl)pyrimidin-5-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 058

At 80°C, a solution of compound 050c (40 mg), 5-oxa-2-azaspiro[3.4]octane hydrochloride (27.40 mg) and N,N-diisopropylethylamine (59.18 mg) in ethanol (1 mL) was stirred and reacted overnight. The crude reaction solution was purified by high performance liquid chromatography (chromatographic column specifications: YMC Triart C18 ExRs, 30 mm × 150 mm, 5 μm, mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; gradient: 45% B to 65% B) to obtain compound 058 (15.24 mg).

LC-MS(ESI): m/z=514.05[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.37 (s, 2H), 8.28 (s, 1H), 7.88 (d, 1H), 7.66-7.60 (m, 1H), 7.50-7.43 (m, 1H) ), 6.25-6.14(m, 1H), 4.03-3.90(m, 4H), 3.79-3.72(m, 2H), 2.45(s, 3H), 2.11-2.05(m, 2H), 1.91-1.82(m ,2H).

Example 6

1-(2-(6,6-difluoro-2-azaspiro[3.3]heptane-2-yl)pyrimidin-5-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 067

At room temperature, diisopropylethylamine (35.51 mg) was added dropwise to a solution of compound 050c (40 mg) and 6,6-difluoro-2-azaspiro[3.3]heptane hydrochloride (31.06 mg) in anhydrous ethanol (2 mL). After the addition was completed, the system was stirred at 80°C overnight. The crude product was purified by HPLC (chromatographic column specifications: YMC Triart C18 ExRs column, 5 μm, 30 mm×150 mm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; gradient: 50% B to 74% B) to obtain compound 067 (13.91 mg).

LC-MS: (ESI, m/z)=533.85[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.37 (s, 2H), 8.29 (s, 1H), 7.89 (d, 1H), 7.66-7.60 (m, 1H), 7.51-7.43 (m, 1H) ), 6.26-6.14(m, 1H), 4.07(s, 4H), 2.90-2.80(m, 4H), 2.45(s, 3H).

Example 7

1-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)-3-(2-(3,3-difluoropyrrolidin-1-yl)pyrimidin-5-yl)urea 068

Compound 050c (25 mg) and 3,3-difluoropyrrolidine (13 mg) were dissolved in anhydrous ethanol (1 mL), heated to 80°C and stirred overnight. The resulting mixture was filtered and the filter cake was washed with anhydrous ethanol (2×1 mL). The crude product was purified by high performance liquid chromatography (chromatographic column specifications: XBridge BEH C18 OBD Prep Column 130, 5 μm, 30 mm×150 mm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution gradient: from 47% B to 63% B in 10 minutes) to obtain compound 068 (4.44 mg).

LC-MS: (ESI, m/z)=507.85[M+H] ⁺

¹ H NMR (400MHz, Methanol-d ₄ ) δ8.28 (s, 1H), 7.36-7.30 (m, 1H), 7.04-6.96 (m, 1H), 6.13-6.04 (m, 1H), 3.82-3.72 (m, 2H), 3.69-3.63 (m, 2H), 2.46-2.32 (m, 5H).

Example 8

(S)-1-(2-aminopyrimidin-5-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 049-1

The first step (S)-N-((5,7-difluoro-3-methylbenzo[b]thiophene-2-yl)methylene)-2-methylpropane-2-sulfinamide 049-1a was added to a tetrahydrofuran (90 mL) solution of 001e (9.1 g) and (S)-2-methylpropane-2-sulfinamide (7.8 g) at room temperature. Tetraethyl titanate (29.4 g) was added. After the addition was completed, the system was heated to 80°C and stirred to react overnight. LCMS monitored the completion of the reaction. The reaction system was cooled to room temperature and quenched with saturated aqueous sodium chloride solution (200 mL), filtered, and the filter cake was washed with ethyl acetate (200 mL). The filtrate was extracted with ethyl acetate (3×200 mL). The organic phases were combined and backwashed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, ethyl acetate/petroleum ether (0-15%) to obtain compound 049-1a (7.5 g).

LC-MS(ESI, m/z)=316.00[M+H] ⁺

Step 2 (S)-N-((S)-1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)-2-methylpropane-2-yl Sulfonamide 049-1b

At room temperature, 049-1a (7.5 g) was dissolved in tetrahydrofuran (80 mL) solution, tetrabutylammonium difluorotriphenyl silicate (19.3 g) was added, and then nitrogen was replaced three times and the temperature was lowered to -60 ° C and stirred for 1 hour. Then (trifluoromethyl) trimethylsilane (13.5 g) was slowly added dropwise. After the addition was completed, the reaction system was stirred at -60 ° C for 1 hour. Then the temperature was raised to -30 ° C and stirred for 1 hour, and then saturated ammonium chloride aqueous solution (100 mL) was added at -30 ° C to quench the reaction. The reaction mixture was extracted with ethyl acetate (3×100 mL), the organic phases were combined, and backwashed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, ethyl acetate/petroleum ether (0-10%) to obtain compound 049-1b (6.5 g).

LC-MS(ESI, m/z)=386.00[M+H] ⁺

Step 3 (S) -1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethane-1-amine 049-1c

At room temperature, a solution of hydrogen chloride in 1,4-dioxane (4M, 80 mL) was added to 049-1b (6.5 g) and stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in water (100 mL), adjusted to pH = 8 with saturated sodium bicarbonate solution, extracted with dichloromethane (2×100 mL), backwashed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure to obtain compound 049-1c (4.5 g).

LC-MS(ESI, m/z)=264.85[M-16] ⁺

Step 4 (S)-1-(2-aminopyrimidin-5-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 049-1

N, N-diisopropylethylamine (367.65 mg) was added to a solution of 049-1c (400 mg, 1.422 mmol, 1 eq) and 049-1d (360.19 mg) in N, N-dimethylformamide (4 mL), and stirred at 60°C for 1 h. The reaction solution was poured into water (50 mL), the reaction mixture was extracted with ethyl acetate (3×30 mL), the organic phase was backwashed with saturated sodium chloride solution (30 mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by Prep_HPLC (chromatographic column specifications: Kinetex 5μm EVO C18, 30mm*150mm; mobile phase A: water (10mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/min; elution gradient: 34% B to 46% B in 7 minutes; detection wavelength: UV 254nm/220nm; retention time (minutes): 6.99) to obtain compound 049-1 (63mg).

LC-MS: (ES, m/z)=417.85[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.21 (s, 2H), 8.11 (s, 1H), 7.85 (d, 1H), 7.66-7.60 (m, 1H), 7.50-7.43 (m, 1H) ), 6.39(s, 2H), 6.24-6.13(m, 1H), 2.45(s, 3H).

(2-aminopyrimidin-5-yl)phenylcarbamate 049-1d

Under nitrogen protection, phenyl chloroformate (853.09 mg) was added to a solution of 2,5-diaminopyrimidine (500 mg) in tetrahydrofuran (5 mL) at room temperature, and the mixture was stirred for 2 hours. The residue was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain compound 049-1d (400 mg).

LC-MS: (ESI, m/z)=231.10[M+H] ⁺

Embodiment 9

(S)-1-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)-3-(1-(2-hydroxyethyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)urea 092-1

The first step 2-(5-bromo-1H-pyrrolo[2,3-b]pyridin-1-yl)ethan-1-ol 092-1b

Potassium hydroxide (2.3 g) was added to a solution of 5-bromo-1H-pyrrolo[2,3-b]pyridine (2 g) in dimethyl sulfoxide (20 mL) at 0°C. After the addition was completed, the system was stirred at room temperature for 1 hour. 2-bromoethanol (1.4 g) was added dropwise at 0°C. After the addition was completed, the system was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate (50 mL), and the reaction mixture was acidified to pH = 6 with hydrochloric acid solution (1N). The reaction mixture was extracted with ethyl acetate (3×100 mL). The organic phases were combined, backwashed with sodium chloride (1×100 mL), and dried over sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0%-50%) to obtain compound 092-1b (400 mg).

LC-MS (ESI, m/z):=240.85[M+H] ⁺

Step 2 (1-(2-hydroxyethyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)carbamic acid tert-butyl ester 092-1c

Under nitrogen protection, at room temperature, cesium carbonate (1.4 g), tris(dibenzylideneacetone)dipalladium (133 mg) and 2-dicyclohexylphosphine-2′, 4′, 6′-triisopropylbiphenyl (138 mg) were added to a solution of 092-1b (350 mg) and tert-butyl carbamate (680 mg) in 1,4-dioxane (2 mL), the nitrogen atmosphere was replaced, and the system was stirred at 100° C. for 2 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic phases were combined, backwashed with sodium chloride (1×50 mL), and dried over sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, ethyl acetate/petroleum ether (0%-40%) to obtain compound 092-1c (200 mg).

LC-MS (ESI, m/z):=278.00[M+H] ⁺

Step 3 2-(5-amino-1H-pyrrolo[2,3-b]pyridin-1-yl)ethan-1-ol 092-1d

At room temperature, hydrogen chloride in 1,4-dioxane (4M, 8 mL) was added to 092-1c (200 mg). After the addition was completed, the system was stirred at room temperature for 1 hour. The residue was concentrated under reduced pressure to obtain the hydrochloride salt of compound 092-1d (120 mg, Crude product).

LC-MS (ESI, m/z):=178.00[M+H] ⁺

Step 4 (S)-1-(1-(5,7-difluoro-3-methylbenzo[b]thiophene-2-yl)-2,2,2-trifluoroethyl)-3-(1-(2-hydroxyethyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)urea 092-1

At room temperature, phenyl chloroformate (59 mg) was added to a solution of compound 049-1c (53 mg) in pyridine (2 mL). After the addition was completed, the system was stirred at room temperature for 2 hours. The residue was concentrated under reduced pressure. The mixture was dissolved in 1,4-dioxane (2 mL), and N,N-diisopropylethylamine (146 mg) and the hydrochloride of 092-1d (40 mg, crude product) were added. After the addition was completed, the system was stirred at 60°C for 2 hours. The residue was concentrated under reduced pressure. The reaction mixture was dissolved with N,N-dimethylformamide (2 mL), and the crude product was purified by HPLC (chromatographic column specifications: Sunfire C18 5 μm, 30 mm*150 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 mL/min; gradient: 45% B to 65% B in 8 minutes; wavelength: 254 nm/220 nm) to obtain compound 092-1 (57 mg).

LC-MS (ESI, m/z):=484.75[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ8.53 (s, 1H), 8.14 (d, 1H), 8.04 (d, 1H), 7.73 (d, 1H), 7.66-7.61 (m, 1H ), 7.52-7.42(m, 2H), 6.36(d, 1H), 6.29-6.18(m, 1H), 4.92-4.87(m, 1H), 4.27-4.22(m, 2H), 3.75-3.68(m ,2H),2.48(s,3H).

Example 10

(S)-1-(2-amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 107-1

First step 6-((diphenylmethylene)amino)-[1,2,4]triazolo[1,5-a]pyridin-2-amine 107-1b

Under nitrogen protection, benzophenone imine (9.36 g), cesium carbonate (45.88 g), tris(dibenzylideneacetone)dipalladium (4.30 g) and 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (5.43 g) were added to a solution of 107-1a (10 g) in dioxane (100 mL) at 25°C, and the mixture was stirred at 100°C for 16 hours. LCMS showed that the product was generated. The mixture was cooled, then filtered and washed with ethyl acetate (100 mL×2). The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography, ethyl acetate/petroleum ether=(0-30%) to obtain compound 107-1b (11.9 g).

LCMS: (ESI, m/z)=314.2[M+H] ⁺

Step 2 (Benzyl (6-((diphenylmethylene)amino)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)carbamate 107-1c

Under nitrogen protection, benzyl chloroformate (2.18 g) and pyridine (1.01 g) were added to a solution of 107-1b (2 g) in dichloromethane (30 mL) at 0°C, and the mixture was stirred at 0°C for 1 hour. LCMS showed that a product was generated. Water was added to the reaction mixture at 0°C. (80mL) was quenched and extracted with dichloromethane (3×60mL). The organic phases were combined, backwashed with saturated brine (1×80mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-70%) to obtain compound 107-1c (0.75g).

LCMS: (ESI, m/z)=448.2[M+H] ⁺

Step 3 (6-amino-[1,2,4]triazolo[1,5-a]pyridin-2-yl)benzyl carbamate 107-1d

To a solution of 107-1c (750 mg) in dichloromethane (12 mL) was added a solution of hydrochloric acid in ethyl acetate (5.88 mL, 4 N) at room temperature, and the reaction system was stirred at room temperature for 2 hours. LCMS showed that a product was generated. The reaction solution was then spin-dried, diluted with water (4 mL), and then adjusted to pH = 10 with a saturated sodium bicarbonate aqueous solution, extracted with dichloromethane (15 mL × 3), the organic phases were combined and spin-dried, and the resulting residue was purified by silica gel column chromatography, dichloromethane/methanol = (10: 1), to give compound 107-1d (410 mg).

LCMS: (ESI, m/z)=284.1[M+H] ⁺

Step 4: Benzyl phenyl[1,2,4]triazolo[1,5-a]pyridine-2,6-diyldiaminobenzyl ester 107-1e

Under nitrogen protection, phenyl chloroformate (208.79 mg) was added to a solution of 107-1d (360 mg) in tetrahydrofuran (8 mL) at room temperature, and the reaction system was stirred at room temperature for 2 hours. LCMS showed that a product was generated. The reaction solution was spin-dried to obtain a crude product of compound 107-1e (534 mg), which was used directly in the next step.

LCMS: (ESI, m/z)=404.2[M+H] ⁺

Step 5 (S)-(6-(3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)carbamic acid benzyl ester 107-1f

Under nitrogen protection, the crude product of 049-1c (290 mg) and 107-1e (498.59 mg) was dissolved in pyridine (6 mL) at room temperature, and the mixture was stirred at 80 ° C for 2 hours. LCMS monitored the formation of products. The reaction solution was diluted with water (30 mL), and then extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated citric acid solution (3×30 mL) and saturated brine (2×30 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (2:1) to obtain compound 107-1f (640 mg).

LCMS: (ESI, m/z)=591.1[M+H] ⁺

Step 6 (S)-1-(2-amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 107-1

Pd/C (106 mg, Pd content 10%) was added to a solution of 107-1f (590 mg) in methanol (10 mL) at room temperature, and the reaction was stirred at 25°C for 1 hour in a hydrogen atmosphere. LCMS showed that the product was generated. The mixture was filtered and washed with methanol (20 mL×2). The filtrate was concentrated and purified by HPLC (Column: Xbridge C18 19×250 mm, 10 μm; mobile phase A: 10 mmol NH ₄ HCO ₃ /H ₂ O, mobile phase B: ACN; gradient: 44% B to 49% B) to obtain compound 107-1 (93.45 mg).

LCMS: (ESI, m/z): 457.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.80-8.78 (m, 1H), 8.58 (s, 1H), 7.89 (d, 1H), 7.66-7.61 (m, 1H), 7.50-7.43 (m , 1H), 7.31-7.22(m, 2H), 6.28-6.18(m, 1H), 5.87(s, 2H), 2.48(s, 3H).

Embodiment 11

1-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)-3-(2-(3-hydroxy-3-methylazetidin-1-yl)pyrimidin-5-yl)urea 052

The first step is 1-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)-3-(2-(3-hydroxy-3-methylazetidin-1-yl)pyrimidin-5-yl)urea 052

050c (40 mg) and 3-methylazetidin-3-ol hydrochloride (20 mg) were dissolved in anhydrous ethanol (2 mL), followed by addition of N, N-diisopropylethylamine (0.25 mL) and heating to 80 ° C and continued stirring overnight. The resulting mixture was cooled to room temperature and then filtered, and the filter cake was washed with anhydrous ethanol (1×1 mL). The crude product was purified by high performance liquid chromatography (chromatographic column specifications: Sunfire C18 5μm, 30*150mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml per minute; gradient: 32% B to 58% B in 8 minutes) to obtain compound 052 (9.13 mg).

LC-MS: (ESI, m/z)=488.00[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.35 (s, 2H), 8.25 (s, 1H), 7.86 (d, 1H), 7.66-7.60 (m, 1H), 7.50-7.42 (m, 1H ), 6.25-6.13(m, 1H), 5.55(s, 1H), 3.87-3.79(m, 4H), 2.45(s, 3H), 1.41(s, 3H).

Example 12

(S)-1-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)-3-(2-(3-hydroxy-3-methylazetidin-1-yl)pyrimidin-5-yl)urea 052-1

The first step (S)-1-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)-3-(2-(3-hydroxy-3-methylazetidin-1-yl)pyrimidin-5-yl)urea 052-1

Dissolve 049-1c and phenyl chloroformate (84 mg) in pyridine (2 mL) and stir at room temperature for 1 hour. Concentrate the reaction mixture under reduced pressure and dissolve it in N, N-dimethylacetamide (2 mL), then add N, N-diisopropylmethylamine (205 mg) and compound 052-1a (96 mg, refer to the synthesis steps in the patent "WO2022265993A1") and heat to 50 ° C and continue stirring for 1 hour. Cool the reaction mixture to room temperature and filter it, and wash the filter cake with N, N-dimethylformamide (2×1 mL). Crude product Compound 052-1 (52.78 mg) was purified by HPLC (chromatographic column specifications: XBridge BEH C18 OBD Prep Column 130, 5 μm, 30 mm*150 mm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; gradient: 30% B to 56% B in 8 minutes).

LC-MS: (ESI, m/z)=487.75[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.35 (s, 2H), 8.23 (s, 1H), 7.84 (d, 1H), 7.65-7.60 (m, 1H), 7.49-7.41 (m, 1H) ), 6.25-6.14(m, 1H), 5.54(s, 1H), 3.87-3.79(m, 4H), 2.45(s, 3H), 1.41(s, 3H).

Example 13

(S)-1-(1-(cyanomethyl)-1H-pyrazol-4-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 109-1

First step (1-(cyanomethyl)-1H-pyrazol-4-yl)phenyl carbamate 109-1b

Under nitrogen protection, phenyl chloroformate (134.81 mg) was added to a solution of 109-1a (100 mg) in tetrahydrofuran (5 mL) at room temperature, and the reaction system was stirred at room temperature for 16 hours. The reaction solution was then spin-dried to obtain compound 109-1b (160 mg, crude product). The crude product was used directly in the next step.

LCMS: (ESI, m/z): 243.1[M+H] ⁺ ;

Step 2 (S)-1-(1-(cyanomethyl)-1H-pyrazol-4-yl)-3-(1-(5,7-difluoro-3-methylbenzo[b]thiophen-2-yl)-2,2,2-trifluoroethyl)urea 109-1

Under nitrogen protection, 109-1b (150 mg) and N,N-diisopropylethylamine (205.49 mg) were added to a solution of 049-1c (150 mg) in dimethyl sulfoxide (6 mL) at room temperature, and the mixture was stirred for 2 hours at room temperature. Water (50 mL) was added to the reaction mixture and extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated brine (2×50 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by prep-HPLC (Waters 2767/QDA, Column: Xbridge C18, 19*250 mm, 10 μm; mobile phase A: 10 mmol NH ₄ HCO ₃ /H ₂ O, mobile phase B: acetonitrile; flow rate: 20 ml/min; gradient: 53% to 53%) to obtain compound 109-1 (99.81 mg).

LCMS: (ESI, m/z): 430.0[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.37 (s, 1H), 7.88 (s, 1H), 7.75 (d, 1H), 7.65-7.60 (m, 1H), 7.51 (d, 1H), 7.49-7.42(m, 1H), 6.24-6.14(m, 1H), 5.39(s, 2H), 2.46(s, 3H).

Using conditions similar to those in the above examples, the compounds listed in Table 1 were prepared, and the structural characterization data of these compounds are listed in Table 1.

Table 1

Biological evaluation

The structure of the control compound used in the test is as follows, and its synthesis steps are shown in WO2022265993A1:

Test Example 1: The purpose of this test is to determine the inhibitory effect of the test compound on PI3Kα and PI3Kα(H1047R) using the ADP-Glo luciferase luminescence detection method.

Test materials and instruments

A. Reagent Information

B. Consumables Information

Experimental Procedure

a. Preparation of reaction buffer

Take 10 ml as an example, prepare and use it on the day of the experiment:

b. Compound Preparation

The test compound was prepared into a 100x concentration stock solution with DMSO, and the compound was graded diluted using a multichannel electronic pipette. 50 nL of the test compound was transferred to a 384-well microvolume assay plate using the automated micropipette system Echo550, where only 50 nL of 100% DMSO solution was added to the wells in the negative control and positive control areas.

c. Experimental steps

The PI3Kα and PI3Kα(H1047R) enzyme solutions and substrate mixed solutions were prepared using reaction buffer. The final concentrations of enzyme and substrate in the reaction solution were as follows:

After the compound transfer is completed, add 2.5μL PI3Ks enzyme solution (compound well and ZPE well) or 2.5μL reaction buffer (HPE well) to each well of the above 384-well experimental plate as shown in the figure, centrifuge at 1000rpm for 1 minute, and then place the 384-well plate in a constant temperature incubator and pre-incubate at 25°C for 10 minutes. After the incubation is completed, add 2.5μL of substrate mixed solution to each well to start the reaction (total reaction volume is 5μL), and place the 384-well plate in a constant temperature incubator for 60 minutes at 25°C. After the reaction, 5 μL of ADP-Glo reagent (Promega, #V9102, thawed and equilibrated to room temperature) was added to stop the reaction. After incubation at 25°C in a constant temperature incubator for 60 minutes, 10 μL of Kinase Detection Substrate (Promega, #V9102, thawed and equilibrated to room temperature) was added to each well, centrifuged and mixed, and then incubated at 25°C in a constant temperature incubator for 30 minutes. The fluorescence value was read on the Envision 2104 multi-function plate reader. The reaction signal value read in each well was the raw data, and the raw data was used to analyze the half-inhibitory concentration of the compound on PI3K kinase.

Experimental Results

This experiment uses XLfit, a software written by IDBS and integrated into the Microsoft Excel environment, for test data processing and analysis. First, calculate the average reaction signal of the positive control well and the negative control well, and then calculate the reaction inhibition rate percentage of each compound well according to the formula "single well inhibition rate = (negative control signal average value-single well signal value)/(negative control signal average value-positive control signal average value)*100%". Then import the concentration and corresponding inhibition rate data into the XLfit software, use the Dose Response One Site 205 model in the software, use the four-parameter method to fit the inhibition rate-concentration curve, and calculate the half-inhibitory concentration (IC ₅₀ value) of the compound.

Experimental Conclusion

The above data show that the representative compounds of the present invention have good PI3Kα inhibitory activity.

Test Example 2 Inhibitory activity of the test compound on E545K mutated MCF7 cells

Materials and reagents

Experimental Procedure

1) Preheat PBS, 0.25% trypsin, and cell culture medium in a 37°C water bath.

2) Observe the cells under a microscope to assess the degree of cell confluence and confirm that there is no bacterial and fungal contamination.

3) Remove the culture medium, wash the cells with 5 mL PBS and aspirate. Add 2 mL 0.25% Trypsin/EDTA reagent to the 100 mm dish. Place the dish in the incubator for a few minutes, or until the cells detach. Add 5 mL fresh cell culture medium, rinse the cells and transfer to a 15 mL centrifuge tube.

4) The collected cells were centrifuged at 1000 rpm for 5 minutes.

5) After centrifugation, discard the supernatant and resuspend the cell pellet in 5 mL of cell culture medium.

6) Take 20μL of resuspended cells and count them. Use Cell Counter Star to add 20μL of cell suspension to 20μL of dye to count cells.

7) Adjust the volume of the suspension using cell culture medium to achieve the desired cell concentration.

8) According to the plate map, transfer 45 μL of cell suspension (cell number see the table below) to each well of a 384-well microplate. As a positive control, add 45 μL of culture medium.

9) Incubate the plate overnight at 37°C/5% _CO2 .

Day 1:

1) Dilute the compound. The stock concentration of the compound is 10 mM, and the starting concentration point of the compound is 10 μM. Use a multichannel pipette to dilute the compound 3 times (5 μl to 10 μL), 10 concentration points.

2) Dilute the compound to 10x the final concentration with growth medium, add 2 ml of the compound at 1000x the final concentration to 198 ml of medium. DMSO% concentration is 1%.

3) Add 5 μL of 10x compound prepared in culture medium to the 384-cell plate (1X).

4) Centrifuge at 1000 rpm for 1 minute and place the cell plate in a cell culture incubator at 37°C and 5% CO2.

Day 8: Readings

1) Add 25 μL/well of CTG reagent to the cell assay plate. Incubate at RT for 10 minutes on a plate shaker at 300 rpm, away from light.

2) Read the luminescent signal on a plate reader (Envision).

Experimental Results

This experiment uses XLfit, a software written by IDBS and integrated into the Microsoft Excel environment, for test data processing and analysis. First, calculate the average reaction signal of the positive control well and the negative control well, and then calculate the reaction inhibition rate percentage of each compound well according to the formula "single well inhibition rate = (negative control signal average value - single well signal value) / (negative control signal average value - positive control signal average value) * 100%". Then import the concentration and corresponding inhibition rate data into the XLfit software, use the Dose Response One Site 205 model in the software, use the four-parameter method to fit the inhibition rate-concentration curve, and calculate the half-inhibitory concentration (IC ₅₀ value) of the compound.

Experimental conclusion:

The compounds of the present invention are significantly better than the control compounds in inhibiting the activity of E545K mutated MCF7 cells. It can be seen that the present invention significantly improves the inhibitory activity of this type of compounds on E545K mutated MCF7 tumor cells through structural optimization, and expands the application scope of this type of compounds.

Test Example 3 Pharmacokinetic study of the test compound in BALB/c female nude mice

Experimental methods

On the day of administration, the test substance was prepared using a solvent formulation of 20% PEG400 + 10% VE-TPGS + 70% HP-β-CD aqueous solution (10% HP-β-CD), and the dosing solution was prepared and ready for use.

The oral gavage PO dose was 100 mg/kg, and the concentration was 10 mg/mL. The animals were fasted overnight, had free access to water, and returned to food 4 hours after administration.

Weigh the animals before administration, calculate the dosage based on the body weight, and administer once by oral gavage on the day of administration. After administration, venous blood was collected at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24h. The blood volume was about 0.03mL/time point, and K2-EDTA was anticoagulated in the blood collection tube. The blood sample was centrifuged within 1h after collection to obtain plasma (centrifugation conditions: 6800g, 6 minutes, 2-8℃), and the samples to be tested were stored in a -80℃ refrigerator before analysis.

Sample preparation for LC-MS/MS assay: 12 μL plasma sample was protein precipitated with 240 μL methanol containing 10 ng/mL internal standard (internal standard is verapamil). The mixture was vortexed for 1 minute and then centrifuged at 4000 rpm for 10 minutes. 200 μL supernatant was transferred to a 96-well plate. 1 μL supernatant was subjected to LC-MS/MS analysis.

The concentration of the test substance in the plasma of BALB/c female nude mice was detected using a validated LC-MS/MS method.

Experimental Results

The pharmacokinetic study of the compounds of the present invention and others in BALB/c female nude mice was determined by the above experiment, and the results are shown in the table below.

Experimental Conclusion

The C _max , AUC and T _1/2 of the compounds of the present invention in BALB/c female nude mice are significantly better than those of the control compounds. It can be seen that the present invention significantly improves the in vivo pharmacokinetic properties of such compounds and enhances the drugability of the compounds through structural optimization.

Test Example 4 Efficacy of the Test Compound in the xxT47D Subcutaneous Tumor Model Established in Balb/c Nude Mice

Experimental methods

Cell culture: Human breast cancer xxT47D cells were cultured in vitro in a suitable culture medium supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 μg/mL streptomycin, and 1.5 μg/mL Blasticidin, and cultured in a 5% CO ₂ incubator at 37°C. When the cell saturation reached 80%-90%, cells in the logarithmic growth phase were collected, counted, and inoculated.

Animals: Balb/c nude mice, 6-8 weeks old, weighing about 18-22 grams. A total of 30 mice (18 plus surplus mice) were provided by Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

Animal husbandry: The experiment can only begin after the animals have been raised in the experimental environment for 3-7 days after arrival. The animals are raised in IVC (independent ventilation system) cages (6 per cage) in an SPF animal room. All cages, bedding and drinking water must be sterilized before use. All experimenters should wear protective clothing and latex gloves when operating in the animal room. The animal information card for each cage should indicate the number of animals in the cage, gender, strain, receipt date, dosing regimen, experiment number, group and start date of the experiment. Cages, feed and drinking water are changed twice a week. The breeding environment and lighting conditions are as follows:

√ Temperature: 20～26℃

√Humidity: 40～70%

Feed ingredients: Feed meets the standards for laboratory animal food identification. The maximum content of pollutants is within the controllable range and is regularly inspected by the manufacturer. High-pressure sterilized drinking water is used for drinking water.

Tumor inoculation: 2-3 days before inoculation, all mice will be subcutaneously implanted with estrogen tablets (0.36mg/60-day release 17β-Estradiol). On the day of inoculation, 0.2mL (10×10 ⁶ cells + Matrigel) xxT47D cells will be subcutaneously inoculated into the right back of the mouse. When the average tumor volume reaches about 150-200mm ³ , grouping and dosing will begin. Weigh the animals and measure the tumor volume before dosing. Randomly group the animals according to the tumor volume (randomized block design). The experimental groups and dosing schedule are shown in Table 2.

Table 2 Grouping and dosing regimen of xxT47D in vivo efficacy animal experiment

Note: 1. N: number of mice in each group; 2. Blank control and compound solvent is 20% PEG400 + 10% VE-TPGS + 70% HP-β-CD aqueous solution (10% HP-β-CD)

Observation: The use and welfare of experimental animals will be carried out in accordance with the rules of the International Association for the Assessment and Accreditation of Laboratory Animal Care (AAALAC). The health status and mortality of animals will be monitored daily. Routine examinations include observing the effects of tumor growth and drug treatment on the daily behavior of animals, such as behavioral activities, food and water intake, weight changes (twice a week), physical signs or other abnormal conditions. The number of deaths and side effects of animals in each group will be recorded based on the number of animals in each group.

Experimental index: The experimental index is to examine whether the tumor growth is inhibited, delayed or cured. The tumor diameter is measured with a vernier caliper twice a week. The calculation formula of tumor volume is: V = 0.5a × ^b2 , a and b represent the long diameter and short diameter of the tumor respectively.

The anti-tumor efficacy of the compound is evaluated by TGI (%) or relative tumor proliferation rate T/C (%). TGI (%) reflects the tumor growth inhibition rate. Calculation of TGI (%): TGI (%) = [1-(average tumor volume at the end of drug administration in a certain treatment group - average tumor volume at the beginning of drug administration in the treatment group)/(average tumor volume at the end of treatment in the solvent control group - average tumor volume at the beginning of treatment in the solvent control group)] × 100%.

Relative tumor proliferation rate T/C (%): The calculation formula is as follows: T/C% = _TRTV / _CRTV × 100% ( _TRTV : RTV of the treatment group; _CRTV : RTV of the negative control group). The relative tumor volume (RTV) was calculated based on the results of tumor measurement. The calculation formula is RTV = _Vt / _V0 , where _V0 is the tumor volume measured at the time of group administration (i.e., d0), _Vt is the tumor volume at a certain measurement, and _TRTV and _CRTV are taken from the same day.

After the experiment, the tumor weight was measured and the percentage of T _weight /C _weight was calculated. T _weight and C _weight represent the tumor weights of the drug administration group and the vehicle control group, respectively.

Termination of the experiment: If the animal's health condition continues to deteriorate, or the tumor volume exceeds 2,000 mm ³ , or there is a serious illness, or pain, it must be euthanized. In the following cases, notify the veterinarian and euthanize:

√ Obvious weight loss, weight loss greater than 20%;

√ Not able to freely access food and water;

√ The experiment was terminated when the average tumor volume of the control group reached 2,000 mm ³ .

√ The animal has the following clinical manifestations and continues to deteriorate:

○ Hair erection

○ Arched back

○ White ears, nose, eyes or feet

○ Shortness of breath

○ Twitching

○Continuous diarrhea

○Dehydration

○ Slow movement

○Voice

Data analysis: T-test was used for comparison between two groups. One-way ANOVA was used for comparison between three or more groups. Two-way ANOVA was used to compare possible differences between different dosing groups. All data were analyzed using Graphpad Prism. p < 0.05 was considered to be significantly different.

Experimental Results

1) Accompanying PK and TGI results

2) Insulin results

Experimental Conclusion

Based on the above results and with reference to Figures 1 and 2, it can be seen that the compound of the new structure obtained by structural optimization of the present invention has a good effect of inhibiting tumor growth at a dose of 100mpk in the xxT47D subcutaneous tumor model established on Balb/c nude mice, and the TGI is better than Alpeilisb when the exposure is lower than Alpelisib. At the same time, during the administration of the present invention, the insulin level did not change significantly, while Alpelisib had a significant increase in insulin after administration, indicating that the present invention is superior to the first-generation PI3Kα drug Alpelisib in terms of safety. In summary, the compound of the present invention has better in vivo efficacy and safety than Alpelisib.

The above is an exemplary description of the implementation of the technical solution of the present disclosure. It should be understood that the protection scope of the present disclosure is not limited to the above implementation. Any modification, equivalent substitution, improvement, etc. made by those skilled in the art within the spirit and principle of the present disclosure shall be included in the protection scope of the claims of this application.

Claims (11)

A compound of formula I and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt;
wherein R ₁ and R ₂ are the same or different and are independently selected from H, deuterium, the following groups which are unsubstituted or optionally substituted by one, two or more R ₁₁ : C _1-12 alkyl, C _1-12 alkyloxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkyloxy, C _3-12 cycloalkyl; each R ₁₁ is the same or different and is independently selected from H, deuterium, halogen, CN, OH, C _1-12 alkyl; each R ₃ is the same or different and is independently selected from H, deuterium, halogen, CN, OH, the following groups which are unsubstituted or optionally substituted with one, two or more R ₃₁ : C _1-12 alkyl, C _1-12 alkyloxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkyloxy, C _3-12 cycloalkyl, 3-14 membered heterocyclyl, C _6-14 aryl, 5-14 membered heteroaryl, N(R ₃₂ )(R ₃₃ ); each R ₃₁ is the same or different and is independently selected from H, deuterium, halogen, CN, C _1-12 alkyl, C _1-12 alkyloxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkyloxy, C _1-12 acyl; R ₃₂ and R ₃₃ are the same or different and are independently selected from H, deuterium, C _1-12 alkyl, S(＝O) ₂ R ₃₁₁ , S(═O)(═NH)R ₃₁₂ ; R ₃₁₁ and R ₃₁₂ are the same or different, and are independently selected from H, deuterium, and C _1-12 alkyl; X is selected from N, NR _x1 or CR _x2 ; R _x1 and R _x2 are the same or different and are independently selected from H, deuterium, halogen, CN, C _1-12 alkyl; Y is selected from O, S or N; Ring A is selected from a C _3-12 carbocyclic ring, a 3-14 membered heterocyclic ring, a C _6-14 aromatic ring, and a 5-14 membered heteroaromatic ring; Each _Ra is the same or different and is independently selected from H, deuterium, halogen, CN, OH, _C1-12 alkyl, _C1-12 alkyloxy, _C3-12 cycloalkyl, halogenated _C1-12 alkyl, halogenated _C1-12 alkyloxy, OH- _C1-12 alkyl; E is absent or selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : _C3-12 cycloalkyl, 3-14 membered heterocyclyl, N( _Re1 )( _Re2 ), _C1-12 alkyl-CN; each _Re is the same or different and is independently selected from H, deuterium, halogen, CN, _C1-12 alkyl-CN, OH, _C1-12 alkyl, _C1-12 alkyloxy, _C3-12 cycloalkyl, halogenated _C1-12 alkyl, halogenated _C1-12 alkyloxy, halogenated _C3-12 cycloalkyl; _Re1 and _Re2 are the same or different and are independently selected from H, deuterium, _C1-12 alkyl, _C1-12 alkyl- _C3-12 cycloalkyl, _C3-12 cycloalkyl- _C1-12 alkyl; m is selected from 0, 1, 2, 3 or 4; n is selected from 0, 1, 2, 3 or 4; p is selected from 0, 1, 2, 3 or 4. The compound according to claim 1 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvent A pharmaceutically acceptable compound, polymorph, metabolite, ester, prodrug or a pharmaceutically acceptable salt thereof, characterized in that R ₁ and R ₂ are the same or different and are independently selected from H, unsubstituted or optionally substituted by one, two or more R ₁₁ : C _1-6 alkyl, halogenated C _1-6 alkyl, C _3-6 cycloalkyl; Preferably, each R ₁₁ is the same or different and is independently selected from H, halogen, C _1-6 alkyl; Preferably, each R ₁₁ is the same or different and is independently selected from H, F, methyl; Preferably, R ₁ and R ₂ are the same or different and are independently selected from H, methyl, ethyl, isopropyl, trifluoromethyl, cyclopropyl, tert-butyl, methylcyclopropyl (such as 1-methyl-cyclopropyl-1-yl), fluorocyclopropyl (such as 1-fluoro-cyclopropyl-1-yl). The compound according to claim 1 or 2 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that each R ₃ is the same or different and is independently selected from H, halogen, CN, the following groups which are unsubstituted or optionally substituted with one, two or more R ₃₁ : C _1-6 alkyl, C _1-6 alkyloxy, halogenated C 1-6 alkyl, halogenated C _1-6 alkyloxy, C _3-6 cycloalkyl, _3-8 membered heterocyclyl, C _6-10 aryl, 5-10 membered heteroaryl, N(R ₃₂ )(R ₃₃ ); Preferably, each R ₃₁ is the same or different and is independently selected from H, CN, C _1-6 alkyl, C _1-6 acyl; Preferably, each R ₃₁ is the same or different and is independently selected from H, C _1-6 alkyl; for example methyl; Preferably, R ₃₂ and R ₃₃ are the same or different and are independently selected from H, C _1-6 alkyl, -S(=O) ₂ -C _1-6 alkyl, -S(=O)(=NH)C _1-6 alkyl; Preferably, R ₃₂ and R ₃₃ are the same or different and are independently selected from H, methyl, S(═O) ₂ CH ₃ , S(═O)(═NH)CH ₃ ; Preferably, each R ₃ is the same or different and is independently selected from H, F, Cl, CN, methyl, methoxy, difluoromethoxy, trifluoromethoxy, methylamino, -NH-S(=O) ₂ CH ₃ , -NH-S(=O)(=NH)CH ₃ , morpholinyl (eg ), tetrahydropyrrolyl (such as ), phenyl, methylpyrazolyl (such as ), cyclopropyl; Preferably, X is selected from N, NR _x1 or CR _x2 ; R _x1 and R _x2 are the same or different and are independently selected from H, halogen, CN, C _1-6 alkyl; Preferably, X is selected from N, NR _x1 or CR _x2 ; R _x1 and R _x2 are the same or different and are independently selected from H, F, Cl, CN, methyl and ethyl. The compound according to any one of claims 1 to 3 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that ring A is selected from a C _3-8 carbocyclic ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring, and a 5-10 membered heteroaromatic ring; Preferably, ring A is selected from a pyrimidine ring, a benzene ring, a pyridine ring, a pyrazine ring, a pyridazine ring, a thiophene ring, a furan ring, a pyrazole ring, Pyrrole ring, thiazole ring, oxazole ring, imidazole ring, triazole ring, quinoline ring, quinazoline ring, pyrrolopyridine ring (such as ), tetrahydroquinoline ring (such as ), cyclopentadienylpyridine ring (such as ), pyrazolopyrimidine ring (such as ), naphthyridine ring (such as ), pyrazolopyridine ring (such as ), imidazopyridine ring (such as ), quinoline ring (such as ), indazole ring (such as ), benzimidazole ring (such as ), triazolopyridine ring (such as ); Preferably, each _Ra is the same or different and is independently selected from H, deuterium, halogen, CN, OH, _C1-6 alkyl, _C1-6 alkyloxy, _C3-6 cycloalkyl, halogenated _C1-6 alkyl, halogenated _C1-6 alkyloxy, OH- _C1-6 alkyl; Preferably, each _Ra is the same or different and is independently selected from H, deuterium, halogen, CN, OH, _C1-3 alkyl, _C1-3 alkyloxy, _C3-6 cycloalkyl, halogenated _C1-3 alkyl, halogenated _C1-3 alkyloxy, OH- _C1-3 alkyl; Preferably, each _Ra is the same or different and is independently selected from H, _C1-6 alkyl, halogenated _C1-6 alkyl, OH- _C1-6 alkyl; for example H, methyl, 2-hydroxyethyl. The compound according to any one of claims 1 to 4 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that E is absent, p is selected from 0; or, E is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Re : 3-10 membered heterocyclic group, N( _Re1 )( _Re2 ); Preferably, E is selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, N( _Re1 )( _Re2 ), _C1-6 alkyl-CN,; Preferably, E is selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : 3-8 membered heterocyclyl, N( _Re1 )( _Re2 ); Preferably, E is selected from the following groups which _are unsubstituted or optionally substituted by one, two or more _Re : _NH2 , azetidinyl, piperidinyl, tetrahydropyrrolyl, morpholinyl, _CH2CH2CN , Preferably, E is selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : _NH2 , azetidinyl, piperidinyl, tetrahydropyrrolyl, morpholinyl, Preferably, each _Re is the same or different and is independently selected from H, halogen, OH, _C1-6 alkyl-CN, _C1-6 alkyl, _C1-6 alkyloxy, halogenated _C1-6 alkyl, halogenated _C1-6 alkyloxy, _C3-6 cycloalkyl; Preferably, each _Re is the same or different and is independently selected from H, F, OH, _CH2- CN, methyl, ethyl, n-propyl, isopropyl, tert-butyl, methoxy, difluoromethoxy, cyclopropyl; Preferably, each _Re is the same or different and is independently selected from H, halogen, OH, _C1-6 alkyl, _C1-6 alkyloxy, halogenated _C1-6 alkyl, halogenated _C1-6 alkyloxy, _C3-6 cycloalkyl; Preferably, each _Re is the same or different and is independently selected from H, F, OH, methyl, ethyl, n-propyl, isopropyl, tert-butyl, methoxy, difluoromethoxy, cyclopropyl; Preferably, _Re1 and _Re2 are the same or different and are independently selected from H, C _1-6 alkyl, C _1-6 alkyl-C _3-8 cycloalkyl, C _3-8 cycloalkyl-C _1-6 alkyl. The compound according to any one of claims 1 to 5 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that the compound has the structure shown below:

Wherein, R ₁ , R ₂ , R ₃ , X, Y, ring A, E, _Ra , _Re , m, n, and p have the definitions as described in any one of claims 1 to 5; Preferably, the compound has the structure shown below:

Wherein, R ₁ , R ₂ , R ₃ , X, Y, ring A, E, _Ra , _Re , m, n, and p have the definitions as described in any one of claims 1 to 5; Preferably, the compound has the structure shown below:

Wherein, E, _Re , _R3 , _Ra , n, and p have the definitions as described in any one of claims 1 to 5; Preferably, the compound represented by formula I may have the structure shown below:
Wherein, _Re and p have the definitions as described in any one of claims 1-5. The compound according to any one of claims 1 to 6 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that the compound has the structure shown below:










A method for preparing the compound according to any one of claims 1 to 7, comprising the following steps:
Wherein, R ₁ , R ₂ , R ₃ , _Ra , _Re , ring A, E, X, Y, m, n, p have the definitions as described in any one of claims 1 to 7; Z is selected from a leaving group, such as halogen, and R is selected from C _6-10 aryl, C _6-10 aryl-C _1-3 alkyl, C _1-3 alkyl-C _6-10 aryl, such as phenyl, tolyl, benzyl. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof. Use of the compound according to any one of claims 1 to 7 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 9 in the preparation of a PI3Kα inhibitor; and/or, use in the preparation of a medicament for preventing and/or treating cancer, such as lung cancer, gastric cancer, endometrial cancer, ovarian cancer, bladder cancer, breast cancer, colon cancer, brain cancer, prostate cancer, skin cancer and/or benign overgrowth syndrome; And/or, use in the preparation of a medicament for preventing and/or treating PIK3CA-associated overgrowth (PROS). A method for preventing and/or treating a disease or condition mediated by PI3Kα, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of at least one compound according to any one of claims 1 to 7 and its racemate, stereoisomer, tautomer, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 9; Preferably, the PI3Kα-mediated disease or symptom is selected from cancer, such as lung cancer, gastric cancer, endometrial cancer, ovarian cancer, bladder cancer, breast cancer, colon cancer, brain cancer, prostate cancer, skin cancer and/or benign overgrowth syndrome; Preferably, the PI3Kα-mediated disease or symptom is PIK3CA-associated overgrowth (PROS).