CN116554150A - Fourth generation EGFR inhibitors - Google Patents

Abstract

The present invention provides a compound as a fourth generation EGFR inhibitor that is a compound or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate, or solvate thereof. The invention also provides pharmaceutical compositions comprising the compounds and their use in the treatment of cancer.

Description

Fourth-generation EGFR inhibitors

Technical Field

The present invention belongs to the field of medicine, and in particular relates to EGFR inhibitors.

Background Art

Lung cancer is one of the most common malignant tumors, with about 1.6 million new cases of lung cancer each year worldwide. It is divided into two types: small cell lung cancer and non-small cell lung cancer (NSCLC), of which non-small cell lung cancer accounts for about 85% of the total number of lung cancers (Nature Reviews Disease Primers, 2015, 1, 15009). Epidermal growth factor receptor (EGFR) is the most common driver gene for non-small cell lung cancer, with a positive rate of 17% in all non-small cell lung cancers, close to 30% to 40% in domestic patients, and as high as about 60% in lung adenocarcinoma.

EGFR is a transmembrane glycoprotein that belongs to the ErbB family of tyrosine kinase receptors. EGFR is abnormally activated by various mechanisms, such as receptor overexpression, mutation, ligand-dependent receptor dimerization, and ligand-independent activation. The continuous activation of its kinase activity initiates downstream signal transduction of cell proliferation, differentiation, and survival. Small molecule inhibitors of EGFR kinase can inhibit the activation of tyrosine kinase, inhibit the proliferation of tumor cells, promote the apoptosis of tumor cells and other biological effects, and are a hot area for lung cancer development.

Osimertinib is a drug developed for first- and second-generation drug-resistant mutations of EGFR (del19 or L858R) accompanied by T790M mutations. It has shown very significant efficacy in clinical practice, but as treatment continues, patients will also develop drug resistance. In 2015 (Nature Medicine, 2015, 21, 560–562), the first report on drug resistance data of 15 patients taking osimertinib was reported, among which EGFR C797S mutation was one of the main mechanisms leading to drug resistance to osimertinib, accounting for about 40%. In addition, the latest literature reports show that among patients with resistance after second-line treatment with osimertinib, 22-25% have C797S mutations (Nature Cancer, 2021, 377-391). Therefore, there is an urgent clinical need to develop new small molecule inhibitors targeting C797S mutations to provide patients with safer and more effective fourth-generation EGFR inhibitors.

Summary of the invention

In the present invention, we used the main mutation types such as del19, del19/T790M/C797S that appeared in the clinic, carried out cell-level evaluation, and verified it at the constructed Ba/F3 cell level, and finally found a series of new chemical entities with strong biological activity. And the inhibition on the wild type is weak, which reflects high selectivity and safety.

In one aspect, the present invention provides a compound, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate or solvate thereof:

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, and optionally a pharmaceutically acceptable excipient.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient, which may also contain other therapeutic agents.

In another aspect, the present invention provides the use of a compound of the present invention in the preparation of a medicament for treating and/or preventing EGFR kinase-mediated diseases.

In another aspect, the present invention provides a method for treating and/or preventing an EGFR kinase-mediated disease in a subject, comprising administering to the subject a compound or composition of the present invention.

In another aspect, the present invention provides a compound of the present invention or a composition of the present invention for use in the treatment and/or prevention of EGFR kinase-mediated diseases.

In a specific embodiment, the disease treated by the present invention includes cancer selected from the group consisting of lung cancer (including non-small cell lung cancer NSCLC, small cell lung cancer (SCLC), lung adenocarcinoma, and lung squamous cell carcinoma).

Other objects and advantages of the present invention will be apparent to those skilled in the art from the following detailed description, examples and claims.

definition

Chemical Definition

Definitions of specific functional groups and chemical terms are described in more detail below.

When a numerical range is listed, each value and subrange within the stated range is intended to be included. For example, "C _1-6 alkyl" includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C _1-6 , C _1-5 , C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-5 , C _{2-4 ,} C _2-3 , C 3-6, C _3-5 , C _3-4 , C _4-6 , C _4-5 , and C _5-6 alkyl.

"C _1-6 alkyl" refers to a straight or branched saturated hydrocarbon group having 1 to 6 carbon atoms. In some embodiments, C _1-4 alkyl and C _1-2 alkyl are preferred. Examples of C _1-6 alkyl include: methyl (C ₁ ), ethyl (C ₂ ), n-propyl (C ₃ ), isopropyl (C 3), n-butyl (C ₄ ), tert-butyl (C ₄ ), sec-butyl (C ₄ ), isobutyl (C ₄ ), n-pentyl (C ₅ ), 3-pentyl (C ₅ ), pentyl (C ₅ ), neopentyl (C ₅ ), 3-methyl-2-butyl (C ₅ ), tert-pentyl (C ₅ ) and n-hexyl (C ₆ ). The term "C _1-6 alkyl" also includes heteroalkyl groups in which one or more (e.g., ₁ , 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). An alkyl group may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, ₁ to 3 substituents, or 1 substituent. Conventional alkyl abbreviations include: Me _{( -CH3 ), Et(-CH2CH3), iPr(-CH(CH3)2), nPr(-CH2CH2CH3), n-Bu(-CH2CH2CH2CH3 ) , or i - Bu} ( _{-CH2CH ( CH3} ) _{2 )} .

"C _1-6 alkylene" refers to a divalent group formed by removing another hydrogen of a C _1-6 alkyl group, and may be substituted or unsubstituted. In some embodiments, C _1-4 alkylene, C _2-4 alkylene, and C _1-3 alkylene are preferred. The unsubstituted alkylene includes, but is not limited to, methylene (-CH ₂ -), ethylene (-CH ₂ CH ₂ -), propylene (-CH ₂ CH ₂ CH ₂ -), butylene (-CH _{2 CH 2 CH 2 CH 2 -), pentylene (-CH 2 CH 2 CH 2 CH 2 CH 2} -), hexylene (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -), and the like. Exemplary substituted alkylene groups, for example, substituted alkylene groups with one or more alkyl(methyl) groups, include, but are not limited to, substituted methylene groups (—CH(CH ₃ )—, —C(CH ₃ ) ₂ —), substituted ethylene groups (—CH(CH ₃ )CH ₂ —, —CH ₂ CH(CH ₃ )—, —C(CH ₃ ) ₂ CH ₂ —, —CH ₂ C(CH ₃ ) _{2 —} ), substituted propylene groups (—CH(CH ₃ )CH ₂ CH ₂ —, —CH ₂ CH(CH ₃ )CH ₂ —, —CH ₂ CH ₂ CH(CH ₃ )—, —C(CH ₃ ) ₂ CH ₂ CH ₂ —, —CH ₂ C(CH ₃ ) ₂ CH ₂ —, —CH ₂ CH ₂ C(CH ₃ ) ₂ —), and the like.

"Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

Therefore, "C _1-6 haloalkyl" refers to the above-mentioned "C _1-6 alkyl" substituted by one or more halogen groups. In some embodiments, C _1-4 haloalkyl is particularly preferred, more preferably C _1-2 haloalkyl. Exemplary haloalkyls include, but are not limited to: -CF ₃ , -CH ₂ F, -CHF ₂ , -CHFCH ₂ F, -CH ₂ CHF ₂ , -CF ₂ CF ₃ , -CCl ₃ , -CH ₂ Cl, -CHCl ₂ , 2,2,2-trifluoro-1,1-dimethyl-ethyl, and the like. The haloalkyl group may be substituted at any available attachment point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

" _C3-10 cycloalkyl" refers to a non-aromatic cyclic hydrocarbon group having 3 to 10 ring carbon atoms and zero heteroatoms. In some embodiments, _C4-10 cycloalkyl, _C3-7 cycloalkyl, _C3-6 cycloalkyl and _C3-5 cycloalkyl are particularly preferred, more preferably _C5-6 cycloalkyl. Cycloalkyl also includes a ring system in which the above cycloalkyl ring is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the cycloalkyl ring, and in such a case, the number of carbons continues to represent the number of carbons in the cycloalkyl system. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl (C ₃ ), cyclopropenyl (C ₃ ), cyclobutyl (C ₄ ), cyclobutenyl (C ₄ ), cyclopentyl (C ₅ ), cyclopentenyl (C ₅ ), cyclohexyl (C _{6 ), cyclohexenyl (C 6 )} , cyclohexadienyl (C ₆ ), cycloheptyl (C ₇ ), cycloheptenyl (C ₇ ), cycloheptadienyl (C ₇ ), cycloheptatrienyl (C ₇ ), etc. The cycloalkyl group may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

"3-10 membered heterocyclyl" refers to a group of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon. In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as the valence permits. In some embodiments, a 4-9 membered heterocyclyl is preferred, which is a 4-9 membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 5-8 membered heterocyclyl is preferred, which is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 3-8 membered heterocyclyl is preferred, which is a 3-8 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms; a 3-7 membered heterocyclyl is preferred, which is a 3-7 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; a 4-7 membered heterocyclyl is preferred, which is a 4-7 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; a 4-6 membered heterocyclyl is preferred, which is a 4-6 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; and a 5-6 membered heterocyclyl is more preferred, which is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms. Heterocyclyl also includes a ring system in which the above-mentioned heterocyclyl ring is fused to one or more cycloalkyl groups, wherein the point of attachment is on the cycloalkyl ring, or a ring system in which the above-mentioned heterocyclyl ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring; and in such a case, the number of ring members continues to represent the number of ring members in the heterocyclyl ring system. Exemplary 3-membered heterocyclyls containing one heteroatom include, but are not limited to, aziridine, oxirane, and thiorenyl. Exemplary 4-membered heterocyclyls containing one heteroatom include, but are not limited to, azetidine, oxirane, and thiorenyl. Exemplary 5-membered heterocyclyls containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to, hexahydrotriazinyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azepanyl, oxepanyl, and thianyl. Exemplary 5-membered heterocyclyl groups fused to a _C6 aryl ring (also referred to herein as 5,6-bicyclic heterocyclyl groups) include, but are not limited to, dihydroindolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl groups fused to a _C6 aryl ring (also referred to herein as 6,6-bicyclic heterocyclyl groups) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like. The heterocyclyl group may be optionally substituted with one or more substituents, for example, substituted with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

"C _6-10 aryl" refers to a monocyclic or polycyclic (e.g., bicyclic) 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having 6-10 ring carbon atoms and zero heteroatoms. In some embodiments, the aryl group has six ring carbon atoms ("C ₆ aryl"; e.g., phenyl). In some embodiments, the aryl group has ten ring carbon atoms ("C ₁₀ aryl"; e.g., naphthyl, e.g., 1-naphthyl and 2-naphthyl). Aryl also includes ring systems in which the above aryl ring is fused to one or more cycloalkyl or heterocyclic groups, and the point of attachment is on the aryl ring, in which case the number of carbon atoms continues to represent the number of carbon atoms in the aryl ring system. Aryl groups may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

"5-10 membered heteroaryl" refers to a group of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur. In heteroaryl containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as the valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings. Heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl or heterocyclic groups, and the point of attachment is on the heteroaryl ring, in which case the number of carbon atoms continues to represent the number of carbon atoms in the heteroaryl ring system. In some embodiments, 5-9 membered heteroaryl is preferred, which is a 5-9 membered monocyclic or bicyclic 4n+2 aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms. In other embodiments, 5-6 membered heteroaryls are particularly preferred, which are 5-6 membered monocyclic or bicyclic 4n+2 aromatic ring systems having ring carbon atoms and 1-4 ring heteroatoms. Exemplary 5 membered heteroaryls containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5 membered heteroaryls containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5 membered heteroaryls containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl), and thiadiazolyl. Exemplary 5 membered heteroaryls containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6 membered heteroaryls containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6 membered heteroaryls containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryls containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryls containing one heteroatom include, but are not limited to, azacycloheptatrienyl, oxacycloheptatrienyl, and thiacycloheptatrienyl. Exemplary 5,6-bicyclic heteroaryls include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indazinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryls include, but are not limited to, naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. A heteroaryl group can be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

Alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl etc. as defined herein are optionally substituted groups.

Exemplary substituents on carbon atoms _include , but are not limited to, halogen, -CN, _-NO2 , -N3, _-SO2H , _-SO3H , -OH, ^-ORaa , -ON( ^Rbb ) ₂ , -N( ^Rbb ) ₂ , -N( ^Rbb ) ₃ ^{+ X-} , -N( ^ORcc ) ^Rbb , -SH, ^-SRaa , ^-SSRcc , -C(= ^O ) ^Raa , _-CO2H , -CHO, -C( ^ORcc ) ₂ , -CO2Raa _, -OC(=O) ^Raa , _-OCO2Raa , -C(=O)N( ^Rbb ) ₂ , -OC( ⁼ O)N( ^Rbb ) ₂ ^{, -NRbbC} (=O) ^Raa _, ^-NRbbCO2Raa , ^-NRbb C(=O)N(R ^bb ) ₂ , -C(=NR ^bb )R ^aa , -C(=NR ^bb )OR ^aa , -OC(=NR ^bb )R ^aa , -OC(=NR ^bb )OR ^aa , -C(=NR ^bb )N(R ^bb ) ₂ , -OC(=NR ^bb )N(R ^bb ) ₂ , -NR ^bb C(=NR ^bb )N(R ^bb ) ₂ , -C(=O)NR ^bb SO ₂ R ^aa , -NR ^bb SO ₂ R ^aa , -SO ₂ N(R ^bb ) ₂ , -SO ₂ R ^aa , -SO ₂ OR ^aa , -OSO ₂ R ^aa , -S(=O)R ^aa , -OS(=O)R ^aa , -Si(R ^aa ) ₃ , -OSi(R ^aa ) ₃ , -C(=S)N(R ^bb ) ₂ , -C(=O)SR ^aa , -C(=S)SR ^aa , -SC(=S)SR ^aa , -SC(=O)SR ^aa , -OC(=O)SR ^aa , -SC(=O)OR ^aa , -P(=O) ₂ R ^aa , -OP(=O) ₂ R ^{a a ,} -P(=O)(R ^aa ) ₂ , -OP(=O)(R ^aa ) ₂ , -OP(=O)(OR ^cc ) ₂ , -P(=O) ₂ N(R ^bb ) ₂ , -OP(=O) ₂ N(R ^bb ) ₂ , -P(=O)(NR ^bb ) ₂ , -OP(=O)(NR ^bb ) ₂ , -NR ^bb P(＝O)( ^ORcc ) ₂ , ^-NRbbP (＝O)( ^NRbb ) ₂ , -P( ^Rcc ) ₂ , -P( ^Rcc ) ₃ , -OP( ^Rcc ) _{2, -OP(Rcc)3} ^, _-B ( ^Raa ) ₂ , -B( ^ORcc ) ₂ , ^-BRaa ( ^ORcc ), alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 ^Rdd groups;

or two geminal hydrogens on a carbon atom are replaced by groups =0, =S, =NN( ^Rbb ) ₂ , = ^NNRbbC (=O) ^Raa , =NNRbbC(=O) _ORaa ^, = ^NNRbbS (=O) ^2Raa , = ^{NRbb or} = ^NORcc ;

each of ^Raa is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, or two ^Ra groups combine to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 ^Rdd groups;

Each of R ^bb is independently selected from: hydrogen, -OH, -OR ^aa , -N(R ^cc ) ₂ , -CN, -C(=O)R ^aa , -C(=O)N(R ^cc ) _2. -CO ₂ R ^aa , -SO ₂ R ^aa , -C(=NR ^cc )OR ^aa , -C(=NR ^cc )N(R ^cc ) ₂ , -SO ₂ N(R ^cc ) ₂ , -SO ₂ R ^cc , -SO ₂ OR ^cc , -SOR ^aa , -C(=S)N(R ^cc ) ₂ , -C(=O)SR ^cc , -C(=S)SR ^cc , -P(=O ) ₂ R ^aa , -P(=O)(R ^aa ) ₂ , -P(=O) ₂ N(R ^cc ) ₂ , -P(=O)(NR ^cc ) ₂ , alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R ^bb groups are combined to form A heterocyclic or heteroaryl ring in which each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl group is independently substituted by 0, 1, 2, 3, 4, or 5 R ^dd group substitution;

each of R ^cc is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R ^cc groups combine to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 R ^dd groups;

Each of R ^dd is independently selected from the group consisting of halogen, -CN, -NO ₂ , -N ₃ , -SO ₂ H, -SO ₃ H, -OH, -OR ^ee , -ON(R ^ff ) ₂ , -N(R ^ff ) ₂ , -N(R ^ff ) ₃ ⁺ X ^- , -N(OR ^ee )R ^ff , -SH, -SR ^ee , -SSR ^ee , -C(＝O)R ^ee , -CO ₂ H, -CO ₂ R ^ee , -OC(＝O)R ^ee , -OCO ₂ R ^ee , -C(＝O)N(R ^ff ) ₂ , -OC(＝O)N(R ^ff ) ₂ , -NR ^ff C(＝O)R ^ee , -NR ^ff CO ₂ R ^ee , -NR ^ff C(＝O)N(R ^ff ) ₂ , -C(=NR ^ff )OR ^ee , -OC(=NR ^ff )R ^ee , -OC(=NR ^ff )OR ^ee , -C(=NR ^ff )N(R ^ff ) ₂ , -OC(=NR ^ff )N(R ^ff ) ₂ , -NR ^ff C(=NR ^ff )N(R ^ff ) ₂ , -NR ^ff SO ₂ R ^ee , -SO ₂ N(R ^ff ) ₂ , -SO ₂ R ^ee , -SO ₂ OR ^ee , -OSO ₂ R ^ee , -S(=O)R ^ee , -Si(R ^ee ) ₃ , -OSi(R ^ee ) ₃ , -C(=S)N(R ^ff ) ₂ , -C(=O)SR ^ee , -C(=S)SR ^ee , -SC(=S)SR ^ee , -P(=O) _2Ree , -P(=O)( ^Ree ) ₂ , -OP(=O)( ^Ree ) ₂ , -OP( ⁼ O)( ^ORee ) ₂ , alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 ^Rgg groups, or two geminal ^Rdd substituents may combine to form =O or =S;

Each of R ^ee is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R ^gg groups;

each of ^Rff is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two ^Rff groups combine to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 ^Rgg groups;

Each of R ^gg is independently: halogen, -CN, -NO ₂ , -N ₃ , -SO ₂ H, -SO ₃ H, -OH, -OC _1-6 alkyl, -ON(C _1-6 alkyl) ₂ , -N(C _1-6 alkyl) ₂ , -N(C _1-6 alkyl) ₃ ⁺ X ^- , -NH(C _1-6 alkyl) ₂ ⁺ X ^- , -NH ₂ (C _1-6 alkyl) ⁺ X ^- , -NH ₃ ⁺ X ^- , -N(OC _1-6 alkyl)(C _1-6 alkyl), -N(OH)(C _1-6 alkyl), -NH(OH), -SH, -SC _1-6 alkyl, -SS(C _1-6 alkyl), -C(＝O)(C _1-6 alkyl), -CO ₂ H, -CO ₂ (C _1-6 alkyl), -OC(＝O)(C 1-6 alkyl), -OCO ₂ (C _1-6 alkyl) _-C ( _{＝NH)2 、} -C(＝NH)O(C _1-6 alkyl), -OC(＝NH)(C _1-6 alkyl), -C(＝NH)NH(C _1-6 alkyl), -C(＝NH) _NH2 、-C(＝NH)O(C _1-6 alkyl), _-OC ( _＝ NH)(C _1-6 alkyl), _-OC (＝NH)OC _1-6 alkyl, _-C (＝NH)N(C _1-6 alkyl) ₂ 、-C(＝NH)NH(C _1-6 alkyl) _, -C(＝NH) _NH2 、-OC(＝NH) _N (C _1-6 alkyl) _-OC (NH)NH(C _1-6 alkyl), -OC(NH)NH ₂ , -NHC(NH) _N (C _1-6 alkyl) ₂ , -NHC(═NH)NH ₂ , -NHSO ₂ (C _1-6 alkyl), -SO ₂ N(C _1-6 alkyl) ₂ , -SO ₂ NH(C _1-6 alkyl), -SO ₂ NH ₂ , -SO ₂ C _1-6 alkyl, -SO ₂ OC _1-6 alkyl, -OSO ₂ C _1-6 alkyl, -SOC _1-6 alkyl, -Si(C _1-6 alkyl) ₃ , -OSi(C _1-6 alkyl) ₃ , -C(═S)N(C _1-6 alkyl) ₂ , C(═S)NH(C 1-6 alkyl), C(═S)NH ₂ , -C(═O)S(C _1-6 alkyl) -C _1-6 alkyl), -C(＝S)SC _1-6 alkyl, -SC(＝S)SC _1-6 alkyl, -P(＝O) ₂ (C _1-6 alkyl), -P(＝O)(C _1-6 alkyl) ₂ , -OP(＝O)(C _1-6 alkyl) ₂ , -OP(＝O)(OC _1-6 alkyl) ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₇ cycloalkyl, C ₆ -C ₁₀ aryl, C ₃ -C ₇ heterocyclyl, C ₅ -C ₁₀ heteroaryl; or two geminal R ^gg substituents may be combined to form ＝O or ＝S; wherein X ^- is a counter ion.

Exemplary substituents on the nitrogen atom include, but are not limited to, hydrogen, -OH, ^-ORaa , -N ^{( Rcc} ) ₂ , -CN, -C( ⁼ O) ^Raa , -C(=O)N( ^Rcc ) ₂ , -CO2Raa, -SO2Raa, -C( ₌ ^NRbb ) ^Raa , -C(= ^{NRcc ) ORaa} , -C(= ^NRcc )N ^{( Rcc} ) ₂ , -SO2N ₍ ^Rcc _{)2, -SO2Rcc, -SO2ORcc, -SORaa, -C(=S)N(Rcc)2 ,} ^-C _{( =} ^O ₎ ^SRcc , -C(=S) ^SRcc , -P(=O) _2Raa , -P(=O)( ^Raa ₎ , -P( ⁼ O) ₂ N( ^Rcc ) ₂ , -P(=O)( ^NRcc ) ₂ , alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two ^Rcc groups attached to the nitrogen atom combine to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 ^Rdd groups, and wherein ^Raa , ^Rbb , ^Rcc and ^Rdd are as described above.

Other definitions

The term "pharmaceutically acceptable salt" as used herein refers to those carboxylates, amino acid addition salts of the compounds of the present invention which are suitable for use in contact with patient tissues within the scope of sound medical judgment, do not produce undue toxicity, irritation, allergic response, etc., are commensurate with a reasonable benefit/risk ratio, and are effective for their intended use, including (where possible) zwitterionic forms of the compounds of the present invention.

"Subjects" for administration include, but are not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or older adults)) and/or non-human animals, e.g., mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and/or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein.

"Disease," "disorder," and "condition" are used interchangeably herein.

Generally, the "effective amount" of a compound refers to an amount sufficient to cause a target biological response. As will be appreciated by those of ordinary skill in the art, the effective amount of the compounds of the invention may vary depending on factors such as the biological target, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and symptoms of the subject. An effective amount includes a therapeutically effective amount and a prophylactically effective amount.

"Combination" and related terms refer to the simultaneous or sequential administration of a compound of the invention and other therapeutic agents. For example, a compound of the invention can be administered simultaneously or sequentially with the other therapeutic agents in separate unit dosage forms, or can be administered simultaneously with the other therapeutic agents in a single unit dosage form.

Example 1

Preparation of key intermediates

Notes on commonly used abbreviations:

Abbreviations: PE = petroleum ether; EA = ethyl acetate; MeOH = methanol; DCM = dichloromethane; DCE = dichloroethane; CH ₃ CN = acetonitrile; 1,4-dioxane = 1,4-dioxane; DMSO = dimethyl sulfoxide; HFIP = hexafluoroisopropanol; DMF = N,N-dimethylformamide; Hex = n-hexane; IPA = isopropanol; NMP = N-methylpyrrolidone; NMO = N-methylmorpholine-N-oxide; TEA = triethylamine; DIEA = diisopropylethylamine; CuI = cuprous iodide; CuCN = cuprous cyanide; triphosgene = triphosgene; p-TsOH = p-toluenesulfonic acid; T ₃ P = 1-propylphosphoric acid cyclic anhydride; TsN ₃ = p-toluenesulfonyl azide; PPA = polyphosphoric acid; BINAP = 1,1'-binaphthyl-2,2'-bisdiphenylphosphine.

Synthesis of intermediates a1-a2, a7-a10

Step 1: In an ice bath, dissolve the raw material 2-amino-3-fluoro-5-iodobenzoic acid methyl ester a1-1 (3.0 g, 10.2 mmol) and copper bromide (225 mg, 1.02 mmol) in 50 mL of acetonitrile, slowly add nitrous acid (1.05 g, 10.2 mmol), dropwise, react at room temperature for 4 hours, and stop the reaction. Add 200 mL of ice water to the reaction solution, extract with ethyl acetate, dry with anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography to obtain a white solid a1-2 (2.1 g), yield: 58%, LCMS: ESI-MS (m/z): 358.8 [M+H] ⁺ .

Step 2: Under nitrogen protection, at -10°C, the intermediate a1-2 (2.1 g, 5.85 mmol) was dissolved in 18 mL of anhydrous THF, and diisobutylaluminum hydride DIBAL-H (17.5 mL, 1 M) was slowly added dropwise, and the temperature was raised to room temperature for 12 hours to stop the reaction. 1 M dilute hydrochloric acid (20 mL) was added to the system, filtered, extracted with ethyl acetate, and the solvent was evaporated under reduced pressure to obtain an oily intermediate a1-3 (1.8 g), LCMS: ESI-MS (m/z): 330.9 [M+H] ⁺ .

Step 3: Dissolve the crude product a1-3 (1.8 g) and triethylamine (1.74 g, 17.2 mmol) in 20 mL of dichloromethane, slowly add methanesulfonic anhydride (1.5 g, 8.61 mmol), react at room temperature for 1 hour, and stop the reaction. Place the reaction solution in an ice bath, add 40 mL of ice water to the system, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate to obtain intermediate a1-4 (2.0 g), which is directly used in the next step. LCMS: ESI-MS (m/z): 408.8 [M+H] ⁺ .

Step 4: In an ice bath, dissolve the raw material 2,2-diethoxyacetamide a1-5 (1.44 g, 9.78 mmol) in 20 mL of anhydrous tetrahydrofuran, slowly add NaH (390 mg, 9.78 mmol), stir for 15 minutes, add the intermediate a1-4 (1.8 g) in the previous step, heat to 50 ° C and react for 2 hours, and stop the reaction. Add 60 mL of ice water to the system, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and separate the crude product by flash column chromatography to obtain intermediate a1-6 (1.2 g), three-step yield: 45%, LCMS: ESI-MS (m/z): 459.9 [M+H] ⁺ .

Step 5: Under nitrogen protection, the intermediate a1-6 (1.2 g, 2.61 mmol), potassium carbonate (1.08 g, 7.83 mmol) and the raw material isopropenylboronic acid pinacol ester a1-7 (460 mg, 2.74 mmol) were dissolved in 11 mL of a mixed solution of 1,4-dioxane and water (v/v, 10/1), and the catalyst Pd(dppf)Cl ₂ (95 mg, 0.13 mmol) was added. After stirring for 5 minutes, the temperature was raised to 80°C for reaction for 5 hours to stop the reaction. 60 mL of ice water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography to obtain intermediate a1-8 (900 mg), yield: 92%, LCMS: ESI-MS (m/z): 374.1 [M+H] ⁺ .

Step 6: Under hydrogen atmosphere (1 atm), the intermediate a1-8 (1.0 g, 2.67 mmol) and palladium carbon (100 mg) were dissolved in 11 mL of ethanol, reacted at room temperature for 12 hours, stopped the reaction, and filtered. The solvent was evaporated under reduced pressure to obtain the intermediate a1-9 (900 mg), yield: 90%, LCMS: ESI-MS (m/z): 376.1 [M+H] ⁺ .

Step 7: Dissolve the intermediate a1-9 (900 mg, 2.39 mmol) in 5 mL of concentrated sulfuric acid, stir for 5 minutes, heat to 50 °C and react for 2 hours to stop the reaction. Slowly pour the reaction solution into 50 mL of ice water, extract with dichloromethane, dry with anhydrous sodium sulfate, filter and concentrate to obtain intermediate a1-10 (300 mg), yield: 44%, LCMS: ESI-MS (m/z): 284.0 [M+H] ⁺ .

Step 8: In an ice bath, dissolve the intermediate a1-9 (250 mg, 0.88 mmol) and diisopropylethylamine DIEA (340 mg, 2.64 mmol) in 5 mL of dichloromethane, slowly add trifluoromethanesulfonic anhydride (320 mg, 1.14 mmol), stir and warm to room temperature for 1 hour, and stop the reaction. Add 30 mL of ice water to the system, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and separate the crude product by flash column chromatography to obtain intermediate a1 (200 mg), yield: 55%, LCMS: ESI-MS (m/z): 415.9 [M+H] ⁺ .

Referring to the synthetic route of compound a1, the following intermediates were synthesized using similar raw materials/skeleton structures.

Synthesis of intermediates a3-a5

Step 1: In an ice bath, dissolve the raw material 1H-4-pyrazole boronic acid pinacol ester a3-1 (25.0 g, 130 mmol) in 300 mL of anhydrous DMF, slowly add NaH (7.7 g, 190 mmol), stir for 5 minutes, add cyclopropylsulfonyl chloride (19.9 g, 140 mmol), react at room temperature for 12 hours, and stop the reaction. Add 1 L of ice water to the system, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate to obtain intermediate a3-2 (35.0 g), yield: 91%, LCMS: ESI-MS (m/z): 299.2 [M+H] ⁺ .

Step 5: Under nitrogen protection, the intermediate a3-2 (27.5 g, 92.2 mmol), cesium carbonate (90.2 g, 280 mmol) and the raw material 2-chloro-4-aminopyrimidine a3-3 (11.9 g, 92.2 mmol) were dissolved in 250 mL of a mixed solution of 1,4-dioxane and water (v/v, 5/1), and the catalyst Pd(dppf)Cl ₂ (3.4 g, 4.6 mmol) was added. After stirring for 5 minutes, the temperature was raised to 90°C for reaction for 10 hours, the reaction was stopped, and the mixture was filtered. 1 L of ice water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography to obtain intermediate a3 (8.7 g), with a yield of 36%, LCMS: ESI-MS (m/z): 266.3 [M+H] ⁺ .

Referring to the synthetic route of compound a3, the following intermediates were synthesized using similar raw materials/skeleton structures.

Synthesis of intermediate a6

Step 1: Under nitrogen protection, the raw material 3-bromothiophene a6-1 (2.0 g, 12.3 mmol), N,N'-dimethyl-1,2-cyclohexanediamine (870 mg, 6.13 mmol), potassium iodide (1.02 g, 6.13 mmol) and the raw material sodium cyclopropanesulfite a6-2 (3.14 g, 24.5 mmol) were added into a 35 mL microwave reactor, and the catalyst [Cu(OTf)] ₂ -toluene (1.27 g, 2.45 mmol) was added. After stirring for 5 minutes, the microwave temperature was raised to 100°C and reacted for 2 hours. The reaction was stopped and filtered. 100 mL of ice water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography to obtain intermediate a6-3 (2.2 g). Yield: 95%, LCMS: ESI-MS (m/z): 189.0 [M+H] ⁺ .

Step 2: In an ice bath, under nitrogen protection, the intermediate a6-3 (1.8 g, 9.56 mmol) and aluminum chloride (1.53 g, 11.5 mmol) were dissolved in 20 mL of dichloromethane, and bromine (1.68 g, 10.5 mmol) was slowly added dropwise, and the temperature was raised to room temperature for 2 hours to stop the reaction. 50 mL of saturated sodium thiosulfate aqueous solution was added to the reaction solution, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography to obtain intermediate a6-4 (1.6 g), yield: 63%, LCMS: ESI-MS (m/z): 266.9 [M+H] ⁺ .

Step 3: Under nitrogen protection, the intermediate a6-4 (1.0 g, 3.74 mmol), potassium acetate (730 mg, 7.48 mmol) and bis(boronic acid) pinacol ester (1.42 g, 5.61 mmol) were dissolved in 10 mL of toluene, and the catalyst Pd(dppf)Cl ₂ (310 mg, 0.37 mmol) was added. After stirring for 5 minutes, the temperature was raised to 80°C for reaction for 10 hours, the reaction was stopped, and the reaction was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography to obtain intermediate a6 (900 mg), with a yield of 77%, LCMS: ESI-MS (m/z): 315.1 [M+H] ⁺ .

Synthesis of intermediate b1

Step 1: Under nitrogen protection, raw material b1-1 (7.0 g, 27.7 mmol) and triethylamine (4.2 g, 41.5 mmol) were dissolved in 40 mL of dichloromethane, and methylsulfonyl chloride (3.21 g, 28.0 mmol) was added, and the mixture was reacted at room temperature for 4 hours. The reaction was stopped, and the solvent was removed under reduced pressure to obtain a crude product b1-2 (10.8 g).

Step 2: Dissolve the crude product b1-2 (10.8 g) and the raw material b1-3 (4.21 g, 27.7 mmol) in 30 mL DMF, add NaH (1.11 g, 27.7 mmol), slowly heat to 80 ° C and react for 12 hours, and monitor the reaction completion by LC-MS. Add 100 mL of saturated saline solution to the system to quench the reaction, extract with dichloromethane, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate to obtain a crude product b1-4 (15 g). LC-MS: ESI-MS (m/z): 388.3 [M+H] ⁺ .

Step 3: Dissolve the crude product b1-4 (15 g) and LiCl (2.52 g, 60.0 mmol) in 40 mL N,N-dimethylacetamide DMA, heat to 150°C for 2 hours, and stop the reaction. Add 100 mL of ice water to the reaction solution, extract with dichloromethane, dry with anhydrous sodium sulfate, filter, concentrate, and separate by flash column chromatography to obtain a yellow solid b1-5 (6.42 g). The total yield of the three steps is 71%. LC-MS: ESI-MS (m/z): 330.1 [M+H] ⁺ .

Step 4: Under hydrogen atmosphere (2 atm), the intermediate b1-5 (6.42 g, 19.5 mmol) and Pd/C (1.28 g, 20%) were mixed in 40 mL of methanol, and 2 mL of trifluoroacetic acid was added. The reaction was allowed to proceed at room temperature for 16 hours, and the reaction was stopped. The mixture was filtered and concentrated to obtain a yellow oil b1 (4.5 g), which was directly used in the next step. LC-MS: ESI-MS (m/z): 164.2 [M+H] ⁺ .

Synthesis of intermediates b2-b5

Step 1: Add intermediate b1-2 (70.0 g, 210 mmol) to a 500 mL reaction bottle, slowly add methylamine isopropanol solution (200 mL, concentration 28%), heat to 70 ° C for 4 hours, and monitor the reaction to be complete. Evaporate the solvent under reduced pressure, separate the crude product by flash column chromatography, and obtain oily intermediate b2-1 (39.0 g), yield: 69%, LCMS: ESI-MS (m/z): 267.4 [M+H] ⁺ .

Step 2: Dissolve the intermediate b2-1 (39.0 g, 150 mmol) and triethylamine (44.5 g, 440 mmol) in 400 mL of dichloromethane, slowly add methanesulfonic anhydride (30.6 g, 180 mmol), react at room temperature for 1 hour, and stop the reaction. Place the reaction solution in an ice bath, add 1 L of ice water to the system, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate to obtain intermediate b2-2 (40 g), which is directly used in the next step reaction. LCMS: ESI-MS (m/z): 345.5 [M+H] ⁺ .

Step 4: Under hydrogen atmosphere (1 atm), the intermediate b2-2 (40 g, 120 mmol) and trifluoroacetic acid (53.0 g, 460 mmol) were dissolved in 200 mL of methanol, and palladium carbon (4.0 g) was slowly added. The reaction was allowed to react for 12 hours at room temperature, and the reaction was stopped, filtered, and the solvent was evaporated under reduced pressure. 100 mL of water was added to the mixture, and impurities were removed by extraction with dichloromethane. The aqueous phase was freeze-dried by a freeze dryer to obtain intermediate b3 (27.0 g), with a yield of 84%, LCMS: ESI-MS (m/z): 179.1 [M+H] ⁺ .

Referring to the synthetic route of compound b2, the following intermediates were synthesized using similar raw materials/skeleton structures.

Synthesis of intermediate c1

Step 1: Under nitrogen protection, intermediate a2 (3.25 g, 8.15 mmol), cesium carbonate (5.31 g, 16.3 mmol) and intermediate a4 (2.0 g, 8.15 mmol) were dissolved in 25 mL 1,4-dioxane, and the catalyst XantphosPd-G ₃ (420 mg, 0.41 mmol) was added. After stirring for 5 minutes, the temperature was raised to 80°C for reaction for 2 hours, the reaction was stopped, and the mixture was filtered. 100 mL of water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography to obtain intermediate c1-1 (2.7 g), yield: 67%, LCMS: ESI-MS (m/z): 493 [M+H] ⁺ .

Step 2: In an ice bath, dissolve the intermediate c1-1 (2.7 g, 5.47 mmol) in 60 mL of hydrogen chloride in 1,4-dioxane (2 M concentration), react at room temperature for 1 hour, and stop the reaction. Add 100 mL of saturated sodium bicarbonate aqueous solution to the system to adjust the pH to about 9, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate to obtain intermediate c1 (2.2 g), which is directly used in the next step. LCMS: ESI-MS (m/z): 409 [M+H] ⁺ .

Synthesis of intermediates C2-C7

Step 1: Under nitrogen protection, the intermediate a8 (2.0 g, 3.96 mmol), sodium tert-butoxide (1.14 g, 11.9 mmol) and the raw material c2-1 (784 mg, 7.92 mmol) were dissolved in 20 mL of toluene, and the catalyst Pd ₂ dba ₃ (733 mg, 0.8 mmol) and the ligand BINAP (498 mg, 0.8 mmol) were added. After stirring for 5 minutes, the temperature was raised to 110°C for reaction for 12 hours, the reaction was stopped, and the mixture was filtered. 100 mL of water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography (PE/EA, 4/1) to obtain the intermediate c2-2 (830 mg), yield: 44%, LCMS: ESI-MS (m/z): 479 [M+H] ⁺ .

Step 2: Under nitrogen protection, the intermediate c2-2 (600 mg, 1.26 mmol), cesium carbonate (1.44 g, 4.41 mmol) and intermediate b2 (640 mg, 2.3 mmol) were dissolved in 6 mL 1,4-dioxane, and the catalyst XantPhosPdG ₃ (120 mg, 0.13 mmol) was added. After stirring for 5 minutes, the temperature was raised to 110°C for reaction for 4 hours, the reaction was stopped, and the mixture was filtered. 100 mL of water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography (PE/EA, 2/1) to obtain intermediate c2 (150 mg), yield: 21%, LCMS: ESI-MS (m/z): 575 [M+H] ⁺ .

Step 3: Dissolve the intermediate c2 (150 mg, 0.26 mmol) in 6 mL of tetrahydrofuran, add TBAF (120 mg, 0.34 mmol), react at room temperature for 1.5 hours, stop the reaction, and filter. Add 100 mL of water to the mixture, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate to obtain intermediate c3 (100 mg), LCMS: ESI-MS (m/z): 419 [M+H] ⁺ .

Referring to the synthetic route of compound c2/c3, the following intermediates were synthesized using similar raw materials/skeleton structures.

Example 2

Preparation of target molecules P1-P5, P13-P14

Step 1: Under nitrogen protection, the intermediate c1 (500 mg, 1.22 mmol) and the raw material P1-1 (350 mg, 1.22 mmol) were dissolved in 5 mL of anhydrous DMF, and NaH (78 mg, 1.95 mmol) was added. After stirring for 5 minutes, the mixture was heated to 80°C for 2 hours, the reaction was stopped, and the mixture was filtered. 30 mL of water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried with anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography to obtain compound P1-2 (270 mg), yield: 39%, LCMS: ESI-MS (m/z): 564 [M+H] ⁺ .

Step 2: In an ice bath, dissolve the intermediate P1-2 (270 mg, 0.48 mmol) in 6 mL of dichloromethane, add 2 mL of trifluoroacetic acid, react at room temperature for 2 hours, and stop the reaction. Add saturated sodium bicarbonate aqueous solution to the system to adjust the pH to about 9, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate to obtain compound P1-3 (150 mg), which is directly used in the next step. LCMS: ESI-MS (m/z): 464 [M+H] ⁺ .

Step 3: Dissolve the intermediate P1-3 (150 mg, 0.32 mmol) and formaldehyde aqueous solution (0.6 mL) in a mixed solution of 12 mL of dichloromethane and tetrahydrofuran (v/v, 1/1), add sodium triacetyl borohydride (750 mg, 3.54 mmol), react at room temperature for 2 hours, and stop the reaction. Add 50 mL of water to the system, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and separate the crude product by flash column chromatography to obtain compound P1-4 (26 mg), yield: 17%, LCMS: ESI-MS (m/z): 478 [M+H] ⁺ .

Step 4: Under nitrogen protection, the compound P1-4 (26 mg, 0.054 mmol), cesium carbonate (53 mg, 0.16 mmol) and intermediate b2 (24 mg, 0.086 mmol) were dissolved in 2 mL 1,4-dioxane, and the catalyst XantphosPd-G ₃ (8.4 mg, 0.008 mmol) was added. After stirring for 5 minutes, the temperature was raised to 110°C for reaction for 4 hours, the reaction was stopped, and the reaction was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography to obtain the target molecule P1 (7.0 mg), with a yield of 22%, LCMS: ESI-MS (m/z): 576 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ10.25 (s, 1H), 9.15 (s, 1H), 8.76 (s, 1H), 8.47 (s, 1H), 8.39 (d, J = 1.5Hz, 1H),8.16(s,1H),7.50(d,J＝7.8Hz,1H),7.21(d,J＝5.1Hz,1H),6.71(d,J＝7.7Hz,1H),5.07(t, J＝7.1Hz,1H),4.69(t,J＝7.8Hz,1H), 4.51–4.46(m,1H),4.25(t,J＝6.6Hz,1H),3.86(t,J＝7.2Hz,1H),3.76(t,J＝6.6Hz,2H),3.63(t,J ＝6.5Hz,1H),2.98(s,3H),2.89–2.83(m,6H),2.38(s,3H),1.44(d,J＝6.0Hz,3H),1.39(dd,J＝8.3, 7.0Hz,6H).

Referring to the synthetic route of compound P1, the following target molecules were synthesized using a similar skeleton structure.

Example 3

Preparation of target molecule P6

Step 1: Under nitrogen protection, the intermediate c1 (400 mg, 0.98 mmol) and the raw material P4-1 (300 mg, 1.1 mmol) were dissolved in 5 mL of anhydrous DMF, and NaH (51 mg, 1.3 mmol) was added. After stirring for 5 minutes, the temperature was raised to 80°C for reaction for 8 hours, the reaction was stopped, and the mixture was filtered. 30 mL of water was added to the mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by flash column chromatography to obtain compound P6-1 (400 mg), yield: 69%, LCMS: ESI-MS (m/z): 592 [M+H] ⁺ .

Step 4: Under nitrogen protection, the compound P6-1 (400 mg, 0.68 mmol), cesium carbonate (774 mg, 2.37 mmol) and intermediate b2 (281 mg, 1.36 mmol) were dissolved in 3 mL 1,4-dioxane, and the catalyst XantphosPd-G ₃ (76 mg, 0.068 mmol) was added. After stirring for 5 minutes, the temperature was raised to 110°C for reaction for 8 hours, the reaction was stopped, and the mixture was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography to obtain compound P6-2 (200 mg), with a yield of 73%, LCMS: ESI-MS (m/z): 690 [M+H] ⁺ .

Step 2: In an ice bath, dissolve the compound P6-2 (200 mg, 0.29 mmol) in 3 mL of dichloromethane, add 2 mL of trifluoroacetic acid, react at room temperature for 2 hours, and stop the reaction. Add saturated sodium bicarbonate aqueous solution to the system to adjust the pH to about 9, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and separate the crude product by HPLC preparative chromatography to obtain the target molecule P6 (100 mg), yield: 59%, LCMS: ESI-MS (m/z): 590 [M+H] ⁺ .

1H NMR (400MHz, DMSO-d ₆ ) δ10.19 (s, 1H), 9.12 (s, 1H), 8.77 (s, 1H), 8.34 (d, J = 5.8Hz, 1H), 8.29 (s, 1H) ),8.09(s,1H),7.47(d,J＝8.0Hz,1H),7.12(d,J＝5.6Hz,1H),6.68(d,J＝8.0Hz,1H),4.66(t,J ＝7.4Hz,1H),4.45(t,J＝6.0Hz,1 H),4.32–4.18(m,2H),3.83(t,J＝7.1Hz,1H),3.62(dd,J＝13.5,6.6Hz,1H),3.11–3.05(m,2H),2.95(s ,3H),2.83(s,3H),2.60(t,J＝11.6Hz,2H),2.08–1.95(m,3H),1.89–1.78(m,2H),1.45–1.32(m,9H).

Example 4

Preparation of target molecules P7, P15-P17

Step 1: Under nitrogen protection, intermediate a1 (200 mg, 0.48 mmol), cesium carbonate (470 mg, 1.44 mmol) and intermediate a3 (130 mg, 0.48 mmol) were dissolved in 5 mL 1,4-dioxane, and catalyst XantphosPd-G ₃ (50 mg, 0.048 mmol) was added. After stirring for 5 minutes, the temperature was raised to 80°C for reaction for 5 hours, the reaction was stopped, and the mixture was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography to obtain compound P7-1 (200 mg), with a yield of 78%, LCMS: ESI-MS (m/z): 531.1 [M+H] ⁺ .

Step 4: Under nitrogen protection, the compound P7-1 (240 mg, 0.45 mmol), cesium carbonate (440 mg, 1.35 mmol) and intermediate b2 (170 mg, 0.63 mmol) were dissolved in 3 mL 1,4-dioxane, and the catalyst XantphosPd-G ₃ (46 mg, 0.045 mmol) was added. After stirring for 5 minutes, the temperature was raised to 110°C for reaction for 5 hours, the reaction was stopped, and the reaction was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography to obtain the target molecule P7 (110 mg), with a yield of 39%, LCMS: ESI-MS (m/z): 629.2 [M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ )δ10.42(s,1H),9.25(s,1H),8.80(s,1H),8.69(s,1H),8.50(s,1H),8.44(d,J＝5.9Hz,1H),7.40(d,J＝15.5Hz,1H),7.22(d,J＝5.3Hz,1H),4.86–4.77( m,1H),4.56(t,J＝7.1Hz,1H),4.14(q,J＝6.8Hz,1H),4.04(t,J＝7.1Hz,1H),3.69–3.58(m,1H),2.95(s,3H),2.86(s,3H),1.47–1.39(m,10H),1.38–1. 24(m,4H).

Referring to the synthetic route of compound P7, the following target molecules were synthesized using a similar skeleton structure.

Example 5

Preparation of target molecules P8-P9

Step 1: Under nitrogen protection, intermediate a2 (100 mg, 0.25 mmol), cesium carbonate (165 mg, 0.5 mmol) and intermediate a5 (59 mg, 0.25 mmol) were dissolved in 3 mL 1,4-dioxane, and catalyst XantphosPd-G ₃ (3 mg, 0.003 mmol) was added. After stirring for 5 minutes, the temperature was raised to 80°C for reaction for 4 hours, the reaction was stopped, and the mixture was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography to obtain compound P8-1 (40 mg), with a yield of 33%, LCMS: ESI-MS (m/z): 482.0 [M+H] ⁺ .

Step 2: Under nitrogen protection, the compound P8-1 (40 mg, 0.08 mmol), cesium carbonate (81 mg, 0.25 mmol) and intermediate b2 (32 mg, 0.11 mmol) were dissolved in 2 mL 1,4-dioxane, and the catalyst XantphosPd-G ₃ (8.5 mg, 0.008 mmol) was added. After stirring for 5 minutes, the temperature was raised to 110°C for reaction for 12 hours, the reaction was stopped, and the reaction was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography to obtain the target molecule P8 (14.7 mg), with a yield of 31%, LCMS: ESI-MS (m/z): 579.6 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ9.13 (s, 1H), 8.56 (s, 1H), 8.39 (d, J = 5.9Hz, 1H), 8.30 (s, 1H), 8.22 (s, 1H) ,7.83(brs,1H),7.46(d,J＝7.9Hz,1H),6.95(d,J＝5.8Hz,1H),6.62(d,J＝8.0Hz,1H),4.60(t,J＝ 7.4Hz,1H),4.41(p ,J＝6.1Hz,1H),4.30(dd,J＝13.5,6.9Hz,1H),4.15(s,2H),3.85(t,J＝7.2Hz,1H),3.65(dt,J＝13.6, 6.8Hz,1H),2.93(s,3H),2.86(s,3H),1.51(d,J＝6.1Hz,3H),1.45–1.40(m,6H),1.23(s,6H).

Referring to the synthetic route of compound P8, the following target molecules were synthesized using a similar skeleton structure.

Example 6

Preparation of target molecules P10-P12

Step 1: In an ice bath, under nitrogen protection, the intermediate c3 (100 mg, 0.24 mmol) and triethylamine (102 mg, 1.0 mmol) were dissolved in 3 mL of anhydrous dichloromethane, trifluoromethanesulfonic anhydride (102 mg, 0.36 mmol) was added, and after stirring for 5 minutes, the temperature was raised to room temperature for 2 hours, and 30 mL of water was added to the reaction solution to quench the reaction. The mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (DCM/MeOH, 20/1) to obtain compound P10-1 (100 mg), yield: 76%, LCMS: ESI-MS (m/z): 551.1 [M+H] ⁺ .

Step 4: Under nitrogen protection, the compound P10-1 (100 mg, 0.18 mmol), cesium carbonate (117 mg, 0.36 mmol) and intermediate a3 (54 mg, 0.20 mmol) were dissolved in 3 mL 1,4-dioxane, and the catalyst XantphosPd-G ₃ (19 mg, 0.02 mmol) was added. After stirring for 5 minutes, the temperature was raised to 110°C for reaction for 4 hours, the reaction was stopped, and the reaction was filtered. The solvent was evaporated under reduced pressure, and the crude product was separated by flash column chromatography (DCM/MeOH, 20/1) to obtain the target molecule P10 (50 mg), with a yield of 42%, LCMS: ESI-MS (m/z): 666.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ10.39(s,1H),9.10(s,1H),8.66(s,1H),8.60(s,1H),8.49(s,1H),8.44( d,J＝5.9Hz,1H),7.16(d,J＝8.3Hz,1H),6.65(d,J＝8.4Hz,1H),4.58(t,J＝7.3Hz,1H),4.53(s, 1H),4.39(dd,J＝11.5,5.7Hz,1H),4.20(s,1H),4.19–4.13(m,1H), 3.98(d,J＝7.5Hz,1H),3.70(dd,J＝14.5,7.4Hz,2H),3.55–3.40(m,1H),3.39–3.35(m,1H),3.30–3.22(m, 1H),2.95(s,3H),2.83(s,3H),2.00–1.92(m,1H),1.82–1.72(m,1H),1.38(d,J＝6.0Hz,3H),1.36–1.31 (m,2H),1.27–1.22(m,2H).

Following the synthetic route of compound P10 and using a similar skeleton structure, the following target molecules were synthesized.

Example 7

Promega CellTiter-Glo reagent was used to detect the effects of small molecule inhibitors on the proliferation of four Ba/F3 cell lines (Ba/F3-EGFR-del19, Ba/F3-FL-EGFR, Ba/F3-EGFR-del19-T790M-C797S and Ba/F3-EGFR-L858R-C797S).

(Ba/F3-EGFR-del19, Ba/F3-EGFR-del19-T790M-C797S and Ba/F3-EGFR-L858R-C797S) culture medium: 1640 medium, 10% FBS, Glutamax and penicillin-streptomycin.

(Ba/F3-FL-EGFR) culture medium: 1640 medium, 10% FBS, Glutamax, 100 ng/mL EGF and penicillin-streptomycin.

The cell line was cultured in an incubator at 37°C and 5% _CO2 . Cells were subcultured regularly and cells in the logarithmic growth phase were used for plating. 95 μL of cell suspension was added to each well of the cell plate, and culture medium without cells (containing 0.1% DMSO) was added to the Min control well. Compound detection cell plate addition: 5 μL of 20× compound working solution was added to the cell culture plate. 5 μL of DMSO-cell culture medium mixture was added to the Max control, and the final DMSO concentration was 0.1%. 5 μL of DMSO-cell culture medium mixture was added to the Max control. The final DMSO concentration was 0.1%.

The culture plate was incubated in a 37°C, 5% CO ₂ incubator for 72 hours. CellTiter-Glo luminescence assay for cell activity. Data analysis:

The cell proliferation inhibition rate (Inhibition Rate) data were processed using the following formula:

Inhibition Rate (Inh%)=100-(RLU _Drug -RLU _Min )/(RLU _Max -RLU _Min )*100%.

Among them: RLU _Drug represents the relative luminescence unit of the cells with the addition of drug, RLU _Min represents the luminescence unit of the culture medium, and RLU _Max represents the relative luminescence unit of the cells with the addition of DMSO.

The inhibition rates corresponding to different concentrations of compounds were calculated in EXCEL, and then the inhibition rate curve was drawn using GraphPad Prism software and related parameters were calculated, including the maximum and minimum inhibition rates of cells and IC ₅₀ values.

Table 1: Antiproliferative effects of representative compounds on BaF3 cells transfected with wild-type EGFR (wt), mutant EGFR (del19) and mutant EGFR (del19/T790M/C797S and Ba/F3-EGFR-L858R-C797S)

N.D. = Not Tested

The above results show that the molecule of the present invention has a good anti-proliferative effect on osimertinib-resistant cell lines (containing C797S mutations), reflecting the effect of the present invention on solving osimertinib-resistant tumors. It also has a good inhibitory effect on the primary cell line of EGFR mutant (EGFR del19), and has a weaker inhibition on the wild type, reflecting the high selectivity of the molecule of the present invention.

Claims (6)

1. A compound, or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from the group consisting of: 2. A pharmaceutical composition comprising the compound of any one of claim 1, or a pharmaceutically acceptable salt, enantiomer, diastereoisomer, solvate, hydrate or isotope thereof variant, and a pharmaceutically acceptable excipient; preferably, it also contains other therapeutic agents. 3. The compound according to any one of claim 1 or its pharmaceutically acceptable salt, enantiomer, diastereoisomer, solvate, hydrate or isotopic variant in the preparation for treatment and/or Or use in medicines for preventing EGFR kinase-mediated diseases. 4. According to claim 3, said EGFR kinase-mediated disease is cancer. 5. According to claim 4, said cancer is lung cancer, further selected from: non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), lung adenocarcinoma, lung squamous cell carcinoma. 6. According to claim 5, said lung cancer is selected from: non-small cell lung cancer NSCLC.