WO2017190637A1 - Fused pyrimidine compound for inhibiting protein tyrosine kinase activity - Google Patents

Abstract

Disclosed are a fused pyrimidine compound as shown in formula (I), a preparation thereof and a pharmaceutical composition comprising the compound, a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotope derivative thereof. The compound can be used for treating and/or preventing protein tyrosine kinase-related diseases, such as cell proliferative diseases, cancers and immunological diseases.

Description

Fused pyrimidine compound for inhibiting protein tyrosine kinase activity Technical field

The invention belongs to the field of medicine. In particular, the present invention relates to fused pyrimidine compounds which have an inhibitory effect on protein tyrosine kinase activity, pharmaceutical compositions containing them, and processes for their preparation and use.

Background technique

The epidermal growth factor receptor (ie, EGFR, ErbB-1 or HER1) is a member of the ErbB receptor family, and the ErbB receptor family includes four closely related receptor tyrosine kinase members: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her3 (ErbB-3) and Her4 (ErbB-4). EGFR is a cell surface receptor for members of the epidermal growth factor family (EGF family) of extracellular protein ligands. Mutations that affect EGFR expression or activity may result in cancer. It has been reported that EGFR is in an dysregulated state in most solid tumors such as lung cancer, breast cancer and brain tumors. It is estimated that 30% of epithelial cancers are associated with mutations, amplification or disorders of EGFR or family members.

Treatments based on inhibition of EGFR by antibody drugs or small molecule inhibitor drugs (eg, gefitinib and erlotinib) have been developed. In the case of non-small cell lung cancer (NSCLC), gefitinib and erlotinib are beneficial for 10% to 40% of patients. However, acquired resistance to gefitinib or erlotinib has become a major clinical problem after a period of treatment. Studies have confirmed that one of the main causes of drug resistance is due to the new mutation in T790M, which is the "gatekeeper" of EGFR. Researchers have then developed inhibitors for T790M, such as BIBW2992, and have shown advantages in clinical trials. However, these inhibitors targeting the T790M mutation of EGFR also have considerable inhibitory activity against wild-type EGFR, which leads to severe toxic side effects that limit its clinical application. Therefore, there is a need to further develop more effective types of selective inhibitors of EGFR that target only mutant but not wild type.

In the case of advanced non-small cell lung cancer with EGFR mutation, although EGFR kinase inhibitors (EGFR-TKI) such as gefitinib and erlotinib have achieved remarkable results, EGFR-TKI was subsequently found to treat non-small cells. Primary or secondary drug resistance in lung cancer is a new challenge for us in the treatment of advanced non-small cell lung cancer. It is necessary to carry out new explorations and find countermeasures.

Third-generation and subsequent EGFR inhibitors include compounds such as AZD9291 and CO-1686, which irreversibly inhibit EGFR and are more efficient in patients with T790M-resistant mutations, but still have an inhibitory effect on wild-type EGFR.

Therefore, it is necessary to further develop new EGFR inhibitors that not only effectively inhibit the T790M mutation, but also have high selectivity for the T790M mutation relative to the wild type.

Summary of the invention

The present invention provides a novel fused pyrimidine compound and a composition comprising the same, and a use thereof, which have better EGFR kinase inhibitory activity and high selectivity for the resistant mutations T790M, L858R and both thereof It can be used to treat, prevent, and alleviate EGFR kinase-mediated diseases.

In this regard, the technical solution adopted by the present invention is as follows:

In a first aspect of the invention, there is provided a fused pyrimidine compound of the formula (I),

among them,

R ^{1 is} selected from H, -OH, halogen, -CN, -NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy base;

L is selected from the group consisting of a bond, NR, O, CR ₂ or S; wherein R is independently selected from H, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy;

R ^{2 is} selected from a C ₁ -C ₆ alkyl group or a 3- to 6-membered heterocyclic group containing 1-2 hetero atoms selected from N and O, wherein the above group is unsubstituted or substituted by 1 to 3 substituents Substituents: halogen, -OH, -CN, -NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, NR ⁶ R ⁷ ; wherein R ⁶ , R ⁷ are each independently selected from C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy or C ₃ -C ₆ carbocyclyl;

R ³ , R ⁴ and R ^{5 are} independently selected from H, halogen, -CN, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl;

Ring A is selected from a C ₃ -C ₆ carbocyclic group, a 3 to 10 membered heterocyclic group, a C ₆ -C ₁₄ aryl group or a C ₅ -C ₁₀ heteroaryl group;

Ring B is selected from the following structures:

Wherein each X ^{1 is} independently selected from the group consisting of C, N, O and S atoms, each X ² and X ^{3 being} independently selected from C and N atoms, wherein the ring containing X ¹ , X ² and X ³ is optionally 1- 3 R ⁸ substituents substituted;

Ring C is selected from a C ₃ -C ₆ carbocyclic group, a 3 to 10 membered heterocyclic group, a C ₆ -C ₁₄ aryl group or a C ₅ -C ₁₀ heteroaryl group, which is optionally substituted by 1 to 3 R ⁸ Base substitution

表示单键或双键；

Represents a single or double bond;

R ^{8 is} independently selected from H, halogen, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy;

Or two R ⁸ on the ring together form -Z _m -; wherein Z is independently selected from CH ₂ , NH, O, S, SO or SO ₂ ; m = 2, 3, 4 or 5;

Or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative thereof.

In another aspect, the invention provides a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable excipient. In a particular embodiment, a compound of the invention is provided in the pharmaceutical composition in an effective amount. In a specific embodiment, the compounds of the invention are provided in a therapeutically effective amount. In a particular embodiment, the compounds of the invention are provided in a prophylactically effective amount.

In another aspect, the invention provides a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable excipient, further comprising other therapeutic agents.

In another aspect, the invention provides a compound, a pharmaceutically acceptable salt, a stereoisomer, a solvate, a hydrate, a crystal form, a prodrug or an isotopic derivative thereof, and other therapeutic agents, and a pharmaceutically acceptable compound, A kit of acceptable carriers, adjuvants or vehicles.

In another aspect, the invention provides a method of treating cancer caused by EGFR (including cancer caused by EGFR mutation, eg, a cancer with a T790M mutation, a L858R mutation, and a L858R/T790M double mutation) in a subject in need thereof A method comprising: administering to a subject an effective amount of a compound of the invention. In a specific embodiment, the EGFR-induced cancer is selected from the group consisting of: non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cancer, Gastrointestinal stromal tumors, leukemia, histiocytic lymphoma, nasopharyngeal carcinoma, etc. In a specific embodiment, the compound is administered orally, subcutaneously, intravenously or intramuscularly. In a specific embodiment, the compound is administered chronically.

Other objects and advantages of the present invention will be apparent to those skilled in the art from the <RTIgt;

definition

Chemical definition

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

When a range of values is recited, each value and sub-range within the range are intended to be included. For example, "C ₁ -C ₆ alkyl" includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₁ -C ₆ , C ₁ -C ₅ , C ₁ -C ₄ , C ₁ - C ₃ , C ₁ -C ₂ , C ₂ -C ₆ , C ₂ -C ₅ , C ₂ -C ₄ , C ₂ -C ₃ , C ₃ -C ₆ , C ₃ -C ₅ , C ₃ -C ₄ C ₄ -C ₆ , C ₄ -C ₅ and C ₅ -C ₆ alkyl.

It will be understood that any of the groups defined below, when described herein, may be substituted with a number of substituents, and the corresponding definitions are within the scope of their inclusion, including substituted groups. Unless otherwise stated, the term "substituted" is as defined below.

"C ₁ -C ₆ alkyl" refers to a straight or branched saturated hydrocarbon group having from 1 to 6 carbon atoms, also referred to herein as "lower alkyl." In some embodiments, a C ₁ -C ₄ alkyl group is particularly preferred. Examples of the alkyl group include, but are not limited to, methyl (C ₁ ), ethyl (C ₂ ), n-propyl (C ₃ ), isopropyl (C ₃ ), n-butyl (C ₄ ), uncle 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-amyl (C ₅ ) and n-hexyl (C ₆ ). Unless otherwise stated, each alkyl group is independently optionally substituted, ie, unsubstituted ("unsubstituted alkyl") or substituted with one or more substituents ("substituted alkyl") For example, 1 to 5 substituents, 1 to 3 substituents or 1 substituent. In some embodiments, the alkyl group is unsubstituted C ₁ -C ₆ alkyl (e.g., -CH _3). In some embodiments, the alkyl group is a substituted C ₁ -C ₆ alkyl.

"C ₁ -C ₆ alkoxy" refers to the group -OR wherein R is a substituted or unsubstituted C ₁ -C ₆ alkyl group. In some embodiments, a C ₁ -C ₄ alkoxy group is particularly preferred. Specific alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentyloxy, N-Hexyloxy and 1,2-dimethylbutoxy.

"Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). In some embodiments, the halo group is F, Cl or Br. In some embodiments, the halogen group is F or Cl. In some embodiments, the halogen group is F.

Thus, "C ₁ -C ₆ haloalkyl" and "C ₁ -C ₆ haloalkoxy" mean the above-mentioned "C ₁ -C ₆ alkyl" and "C ₁ -C ₆ alkoxy", which are one or Multiple halogen groups are substituted. In some embodiments, a C ₁ -C ₄ haloalkyl group is particularly preferred, more preferably a C ₁ -C ₂ haloalkyl group. In some embodiments, a C ₁ -C ₄ haloalkoxy group is particularly preferred, more preferably a C ₁ -C ₂ haloalkoxy group. Exemplary haloalkyl groups include, but the 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. Exemplary haloalkoxy groups include, but are not limited to, -OCH ₂ F, -OCHF ₂ , -OCF ₃ , and the like.

"C ₃ -C ₆ carbocyclyl" refers to a non-aromatic cyclic hydrocarbon group having 3 to 6 ring carbon atoms and zero heteroatoms. In some embodiments, a C ₅ -C ₆ carbocyclic group is preferred. Carbocyclyl also includes ring systems wherein the above carbocyclyl ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the carbocyclic ring, and in such cases, the number of carbons continues to be represented The number of carbons in the carbocyclic system. Exemplary such carbocyclic 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 ), cyclopentadienyl (C ₅ ), cyclohexyl (C ₆ ), cyclohexenyl (C ₆ ), cyclohexadienyl (C ₆ ), etc. Wait. Unless otherwise specified, each of the carbocyclic groups is independently optionally substituted, ie, unsubstituted ("unsubstituted carbocyclyl") or substituted with one or more substituents ("substituted carbocyclic ring"base"). In some embodiments, the carbocyclic group is unsubstituted C ₃ -C ₆ carbocyclyl. In some embodiments, the carbocyclyl is substituted with C ₃ -C ₆ carbocyclyl.

The "3- to 10-membered heterocyclic group" means a group of a 3- to 10-membered non-aromatic ring system having a ring carbon atom and 1 to 4 ring hetero atoms, wherein each hetero atom is independently selected from nitrogen, oxygen, Sulfur, boron, phosphorus and silicon. In the heterocyclic group 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 3- to 6-membered heterocyclic group is particularly preferred, which is a 3- to 6-membered non-aromatic ring system having a ring carbon atom and 1 to 3 ring heteroatoms; more preferably a 5- to 6-membered heterocyclic ring. a group which is a 5 to 6 membered non-aromatic ring system having a ring carbon atom and 1 to 3 ring hetero atoms. Unless otherwise stated, each of the heterocyclic groups is independently optionally substituted, ie, unsubstituted ("unsubstituted heterocyclic") or substituted with one or more substituents ("substituted hetero Ring base"). In some embodiments, the heterocyclyl is an unsubstituted 3-10 membered heterocyclyl. In some embodiments, a heterocyclic group is a substituted 3-10 membered heterocyclyl. The heterocyclic group further includes a ring system in which the above heterocyclic ring is fused to one or more carbocyclic groups, wherein the point of attachment is on the carbocyclic ring, or wherein the above heterocyclic ring is bonded to one or more aryl groups or A heteroaryl fused ring system wherein the point of attachment is on a heterocyclyl ring; and in such cases, the number of ring members continues to indicate the number of ring members in the heterocyclyl ring system. Exemplary 3-membered heterocyclic groups containing one hetero atom include, but are not limited to, aziridine, oxacyclopropane, thicyclopropyl. Exemplary 4-membered heterocyclic groups containing one hetero atom include, but are not limited to, azetidinyl, oxetanyl and thietane. Exemplary 5-membered heterocyclic groups containing one hetero atom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2, 5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolyl, oxathiolanyl, oxathiolanyl (1,2-oxothio) Cyclopentenyl, 1,3-oxathiolanyl, dithiolanyl, dihydropyrazolyl, dihydroimidazolyl, dihydrothiazolyl, dihydroisothiazolyl, dihydrogen Oxazolyl, dihydroisoxazolyl, dihydrooxadiazolyl and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolyl, and thiadiazolyl. Exemplary 6-membered heterocyclic groups containing one hetero atom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridyl and thiacyclohexane. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dihydropyrazinyl, piperazinyl, morpholinyl, dithianyl, dioxoalkyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to, hexahydrotriazinyl. Exemplary 7-membered heterocyclic groups containing one hetero atom include, but are not limited to, azepanyl, oxaheptyl and thiaheptanyl. Exemplary 8-membered heterocyclic groups containing one hetero atom include, but are not limited to, azacyclooctyl, oxacyclooctyl, and thicyclooctyl. Exemplary 5-membered heterocyclic groups (also referred to herein as 5,6-bicyclic heterocyclyl) fused to a _C6 aryl ring include, but are not limited to, indanyl, isoindoline , dihydrobenzofuranyl, dihydrobenzothiophenyl, benzoxazolinone, and the like. Exemplary 6-membered heterocyclic groups fused to a C ₆ aryl ring (also referred to herein as 6,6-bicyclic heterocyclyl) include, but are not limited to, tetrahydroquinolyl, tetrahydroisoquinolinyl, and many more.

"C ₆ -C ₁₄ aryl" refers to a monocyclic or polycyclic (eg, bicyclic or tricyclic) 4n+2 aromatic ring system having 6 to 14 ring carbon atoms and zero heteroatoms (eg, having A group of 6, 10 or 14 π electrons shared in a ring arrangement. In some embodiments, an aryl group having six ring carbon atoms ( "C ₆ aryl"; e.g., phenyl). In some embodiments, an aryl group has ten ring carbon atoms (" _C10 aryl"; for example, naphthyl, eg, 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has fourteen ring carbon atoms (" _C14 aryl"; for example, fluorenyl). In some embodiments, a C _6-10 aryl group is particularly preferred, more preferably a C ₆ aryl group. The aryl group also includes a ring system in which the above aryl ring is fused to one or more carbocyclic or heterocyclic groups, and the point of attachment is on the aryl ring, in which case the number of carbon atoms continues to be represented. The number of carbon atoms in the aryl ring system. Unless otherwise stated, each of the aryl groups is independently optionally substituted, that is, unsubstituted ("unsubstituted aryl") or substituted with one or more substituents ("substituted aryl"). In some embodiments, the aryl group is an unsubstituted C ₆ -C ₁₄ aryl group. In some embodiments, the aryl group is a substituted C ₆ -C ₁₄ aryl group.

"C ₅ -C ₁₀ heteroaryl" means a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms (for example, having a ring-like arrangement) a group of 6 or 10 π electrons, wherein each heteroatom is independently selected from the group consisting of nitrogen, oxygen, and sulfur. In a heteroaryl group 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 systems may include one or more heteroatoms in one or both rings. Heteroaryl also includes ring systems wherein the above heteroaryl ring is fused to one or more carbocyclic or heterocyclic groups, and the point of attachment is on the heteroaryl ring, in this case a carbon atom The number continues to indicate the number of carbon atoms in the heteroaryl ring system. In some embodiments, a C ₅ -C ₆ heteroaryl group is particularly preferred, which is a 5-6 membered monocyclic or bicyclic 4n+2 aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms. . Unless otherwise stated, each of the heteroaryl groups is independently optionally substituted, ie, unsubstituted ("unsubstituted heteroaryl") or substituted with one or more substituents ("substituted heteroaryl"base"). In some embodiments, the heteroaryl is an unsubstituted 5-10 membered heteroaryl. In some embodiments, the heteroaryl is a substituted 5-10 membered heteroaryl. Exemplary 5-membered heteroaryl groups containing one hetero atom include, but are not limited to, pyrrolyl, furyl and thienyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one hetero atom include, but are not limited to, pyridyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one hetero atom include, but are not limited to, azepandinyl, oxepanethylene, and thiephenylene. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, mercapto, isodecyl, oxazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuranyl , benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzooxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, Pyridazinyl and fluorenyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, acridinyl, quinolyl, isoquinolinyl, fluorenyl, quinoxalinyl, pyridazinyl and quinazolinyl .

Other definitions

The term "pharmaceutically acceptable salt" means that, within the scope of sound medical judgment, it is suitable for contact with tissues of humans and lower animals without excessive toxicity, irritation, allergies, etc., and with reasonable benefits/dangers. Those salts that are proportionate. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., pharmaceutically acceptable salts as described in detail in J. Pharmaceutical Sciences (1977) 66: 1-19. Pharmaceutically acceptable salts of the compounds of the invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino and inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or salts with organic acids, such as acetic acid, oxalic acid, Maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or a salt formed using methods used in the art, for example, an ion exchange method. Other pharmaceutically acceptable salts include: adipic acid salts, alginate, ascorbate, aspartate, besylate, benzoate, disulfate, borate, butyrate, camphor Acid salt, camphor sulfonate, citrate, cyclopentanoate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, gluconate, glycerol Phosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate , malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate Salt, pectin ester, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, Thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Pharmaceutically acceptable salts derived from suitable bases include the alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts, and the like. Further pharmaceutically acceptable salts, if appropriate, include non-toxic ammonium salts, quaternary ammonium salts and amine cations formed using counterions, counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, Nitrate, lower alkyl sulfonate and aryl sulfonate.

"Subjects" for administration include, but are not limited to, humans (ie, males or females of any age group, eg, pediatric subjects (eg, infants, children, adolescents) or adult subjects (eg, young Adults, middle-aged adults or older adults) and/or non-human animals, for example, mammals, for example, primates (eg, 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.

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

The term "treatment" as used herein, unless otherwise indicated, includes the effect of a subject having a particular disease, disorder, or condition that reduces the severity of the disease, disorder, or condition, or delays or slows the disease, disorder. Or the development of a condition ("therapeutic treatment"), but also the effect that occurs before the subject begins to have a particular disease, disorder or condition ("prophylactic treatment").

Generally, an "effective amount" of a compound refers to an amount sufficient to cause a target biological response. As will be understood by one of ordinary skill in the art, an effective amount of a compound of the invention can vary depending on, for example, the biological target, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age of the subject. Health conditions and symptoms. Effective amounts include therapeutically and prophylactically effective amounts.

Unless otherwise indicated, a "therapeutically effective amount" of a compound as used herein is in the course of treating a disease, disorder or condition. To provide or limit the likelihood of a therapeutic benefit, or to delay or minimize one or more symptoms associated with a disease, disorder, or condition. A therapeutically effective amount of a compound refers to the amount of a therapeutic agent used alone or in combination with other therapies that provides a therapeutic benefit in the treatment of a disease, disorder or condition. The term "therapeutically effective amount" can include an amount that improves overall treatment, reduces or avoids the symptoms or causes of a disease or condition, or enhances the therapeutic efficacy of other therapeutic agents.

A "prophylactically effective amount" of a compound, as used herein, is an amount sufficient to prevent a disease, disorder, or condition, or a quantity sufficient to prevent one or more symptoms associated with a disease, disorder, or condition, or to prevent disease, unless otherwise stated. The number of relapses of a disorder or condition. A prophylactically effective amount of a compound refers to the amount of a therapeutic agent used alone or in combination with other agents that provides a prophylactic benefit in the prevention of a disease, disorder or condition. The term "prophylactically effective amount" can include an amount that improves the overall amount of prevention, or enhances the prophylactic efficacy of other prophylactic agents.

"Combination" and related terms mean the simultaneous or sequential administration of a therapeutic agent of the invention. For example, a compound of the invention may be administered simultaneously or sequentially with another therapeutic agent in separate unit dosage forms, or together with another therapeutic agent in a single unit dosage form.

"Cancer-induced cancer" refers to a cancer characterized by an unsuitable high expression of the EGFR gene or a mutation of the EGFR gene that alters the biological activity of the EGFR nucleic acid molecule or polypeptide. Cancers caused by EGFR can occur in any tissue including brain, blood, connective tissue, liver, mouth, muscle, spleen, stomach, testes, and trachea. Cancers caused by EGFR include, but are not limited to, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, gastrointestinal stromal tumor, leukemia , tissue cell lymphoma, nasopharyngeal cancer.

An "EGFR mutation" or "EGFR mutant" includes one or more deletions, substitutions or additions in the amino acid or nucleotide sequence of the EGFR protein or EGFR coding sequence. The EGFR mutation may also include one or more deletions, substitutions or additions, or fragments thereof, so long as the mutant retains or increases tyrosine kinase activity relative to wild-type EGFR. In a particular EGFR mutation, the kinase or phosphorylation activity can be increased or decreased relative to wild-type EGFR (eg, at least 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70). %, 80%, 90% or even 100%). Exemplary EGFR mutations include, but are not limited to, the T790M mutation, the L858R mutation, and the L858R/T790M double mutation.

detailed description

Compound

As used herein, "a compound of the invention" refers to a compound of formula (I) below, a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative thereof.

In one embodiment, the invention relates to a compound of formula (I):

among them,

R ^{1 is} selected from H, halogen, -OH, -CN, -NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy base;

L is selected from the group consisting of a bond, NR, O, CR ₂ or S; wherein R is independently selected from H, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy;

R ^{2 is} selected from a C ₁ -C ₆ alkyl group or a 3- to 6-membered heterocyclic group containing 1-2 hetero atoms selected from N and O, wherein the above group is unsubstituted or substituted by 1 to 3 substituents Substituents: halogen, -OH, -CN, -NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, NR ⁶ R ⁷ ; wherein R ⁶ , R ⁷ are each independently selected from C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy or C ₃ -C ₆ carbocyclyl;

R ³ , R ⁴ and R ^{5 are} independently selected from H, halogen, -CN, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl;

Ring A is selected from a C ₃ -C ₆ carbocyclic group, a 3 to 10 membered heterocyclic group, a C ₆ -C ₁₄ aryl group or a C ₅ -C ₁₀ heteroaryl group;

Ring B is selected from the following structures:

Wherein each X ^{1 is} independently selected from the group consisting of C, N, O and S atoms, each X ² and X ^{3 being} independently selected from C and N atoms, wherein the ring containing X ¹ , X ² and X ³ is optionally 1- 3 R ⁸ substituents substituted;

Ring C is selected from a C ₃ -C ₆ carbocyclic group, a 3 to 10 membered heterocyclic group, a C ₆ -C ₁₄ aryl group or a C ₅ -C ₁₀ heteroaryl group, which is optionally substituted by 1 to 3 R ⁸ Base substitution

表示单键或双键；

Represents a single or double bond;

R ^{8 is} independently selected from H, halogen, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy;

Or two R ⁸ on the ring together form -Z _m -; wherein Z is independently selected from CH ₂ , NH, O, S, SO or SO ₂ ; m = 2, 3, 4 or 5;

Or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative thereof.

Preferably, in this embodiment, R ^{1 is} independently selected from halogen, -OH, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ Haloalkoxy. More preferably, R ^{1 is} independently selected from halogen, -OH, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy. More preferably, R ¹ is a C ₁ -C ₆ alkoxy group such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, N-pentyloxy, n-hexyloxy and 1,2-dimethylbutoxy. More preferably, R ¹ is a C ₁ -C ₄ alkoxy group such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy and sec-butoxy. More preferably, R ¹ is a halogen such as Cl or Br. More preferably, R ¹ is Cl or methoxy. Most preferably, R ¹ is methoxy.

Preferably, in this embodiment, L is selected from the group consisting of a bond, NR, O, CR ₂ , or S, wherein R is selected from H or C ₁ -C ₆ alkyl. More preferably, L is selected from NR, O, or S, wherein R is selected from C ₁ -C ₆ alkyl. Most preferably, L is selected from NR and R is a methyl group.

Preferably, in this embodiment, R ^{2 is} selected from a C ₁ -C ₆ alkyl group substituted with _{1 to} 3 substituents or a 3 to 6 membered heterocyclic ring having 1 to 2 hetero atoms selected from N and O. And wherein the substituent is halogen, -OH, -CN, -NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, NR ⁶ R ⁷ . More preferably, R ^{2 is} selected from C ₁ -C ₆ alkyl substituted with _{1 to} 3 substituents, wherein the substituent is NR ⁶ R ⁷ . More preferably, R ^{2 is} selected from a substituent-substituted C ₁ -C ₄ alkyl group, wherein the substituent is NR ⁶ R ⁷ . Most preferably, R ^{2 is} selected from the group consisting of a substituent substituted ethyl group wherein the substituent is N(CH ₃ ) ₂ .

Preferably, in this embodiment, R ³ , R ⁴ and R ^{5 are} independently selected from H, halogen or C ₁ -C ₆ alkyl. More preferably, R ³ , R ⁴ and R ⁵ are both H.

Preferably, in this embodiment, Ring A is selected from a C ₅ -C ₆ carbocyclic group, a 5 to 6 membered heterocyclic group, a C ₆ aryl group or a C ₅ -C ₆ heteroaryl group. More preferably, Ring A is selected from a 5 to 6 membered heterocyclic group or a C ₅ -C ₆ heteroaryl group. More preferably, ring A is selected from a C ₅ -C ₆ heteroaryl group. Most preferably, Ring A is selected from C ₅ heteroaryl groups.

In a preferred embodiment of Ring A, Ring A has the following structure:

among them,

Each Y ^{1 is} independently selected from C, N, O or S atoms, and each Y ^{2 is} independently selected from a C or N atom, which is optionally substituted with an R ⁹ substituent selected from the group consisting of H, halogen, -CN, C _1- C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy or C ₁ -C ₆ alkylsulfonyl, wherein said C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy or C ₁ -C ₆ alkylsulfonyl optionally substituted by one or more -OH .

In a more preferred embodiment of the above technical solution, Ring A has the following structure:

Wherein R ^{9 is} independently selected from H, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy or C ₁ -C ₆ alkyl a sulfonyl group, wherein the C ₁ -C ₆ alkyl group, a C ₁ -C ₆ haloalkyl group, a C ₁ -C ₆ alkoxy group, a C ₁ -C ₆ haloalkoxy group or a C ₁ -C ₆ alkylsulfonyl group The ground is replaced by one or more -OH.

In a more preferred embodiment of the above technical solution, Ring A has the following structure:

Wherein R ⁹ is independently selected from H, C ₁ -C ₆ alkyl, substituted with one or more -OH, C ₁ -C ₆ alkyl.

Preferably, in this embodiment, Ring B has the following structure:

Wherein each X ^{1 is} independently selected from the group consisting of C, N, O and S atoms, each X ² and X ^{3 being} independently selected from C and N atoms, wherein the ring containing X ¹ , X ² and X ³ is optionally 1- 3 R ⁸ substituents substituted;

Ring C is selected from a C ₅ -C ₆ carbocyclic group, a 5 to 6 membered heterocyclic group, a C ₆ -C ₁₀ aryl group or a C ₅ -C ₆ heteroaryl group, which is optionally substituted with from 1 to 3 R ⁸ Base substitution

表示单键或双键；

Represents a single or double bond;

R ^{8 is} independently selected from H, halogen, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy;

Or the two R ⁸ on the ring together form -(CH ₂ ) _m -, -NH(CH ₂ ) _n -, -O(CH ₂ ) _n -, -S(CH ₂ ) _n -, -SO(CH _{2 n} -, -SO ₂ (CH ₂ ) _n -, -CH ₂ NH(CH ₂ ) _p -, -CH ₂ O(CH ₂ ) _p -, -CH ₂ S(CH ₂ ) _p -, -CH ₂ SO(CH ₂ ) _p - or -CH ₂ SO ₂ (CH ₂ ) _p -; wherein m = 2, 3, 4 or 5; n = 1, 2 or 3; p = 1 or 2.

In a more preferred embodiment of the above technical solution, X ¹ is a C atom. In a more preferred embodiment of the above technical solution, X ¹ is an N atom. In a more preferred embodiment of the above technical solution, X ¹ is an O atom. In a more preferred embodiment of the above technical solution, X ¹ is an S atom.

In a more preferred embodiment of the above technical solution, X ² is a C atom. In a more preferred embodiment of the above technical solution, X ² is an N atom.

In a more preferred embodiment of the above technical solution, X ³ is a C atom. In a more preferred embodiment of the above technical solution, X ³ is an N atom.

In a more preferred embodiment of the above technical scheme, Ring C is selected from a C ₅ -C ₆ carbocyclic group, a 5- to 6-membered heterocyclic group, a C ₆ aryl group or a C ₅ -C ₆ heteroaryl group. In a more preferred embodiment of the above technical scheme, ring C is selected from the group consisting of cyclopentadienyl, cyclohexadienyl, phenyl, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, dihydropyridyl, pyrimidine Base, pyrazinyl, dihydropyrazinyl.

In a more preferred embodiment of the above technical solution, R ^{8 is} independently selected from the group consisting of H, F, oxo and C ₁ -C ₆ alkyl. In a more preferred embodiment of the above technical solution, two R ⁸ on the ring together form -(CH ₂ ) ₂ -, -(CH ₂ ) ₃ -, -OCH ₂ -, -O(CH ₂ ) ₂ - Or -CH ₂ OCH ₂ -.

In a more preferred embodiment of the above technical solution, the ring B has the following structure:

Wherein each ring is optionally substituted with from 1 to 3 R ⁸ substituents, and R ⁸ , X ¹ and X ² are as defined above.

In a more preferred embodiment of the above technical solution, two R ⁸ on the same ring together form -Z _m -. In a more preferred embodiment of the above technical solution, two R ⁸ on different rings together form -Z _m -.

In a more preferred embodiment of the above technical solution, the ring B has the following structure:

Wherein each ring is optionally substituted with 1-2 R ⁸ substituents, and R ⁸ , Z, m, X ¹ and X ² are as defined above.

More preferably, in some embodiments, Ring B is selected from the group consisting of:

Wherein R ^{8 is} as defined above.

In a more specific embodiment, the invention relates to the following compounds:

The compounds of the invention may include one or more asymmetric centers, and thus may exist in a variety of stereoisomeric forms, for example, enantiomeric and/or diastereomeric forms. For example, the compounds of the invention may be in the form of individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, A racemic mixture and a mixture rich in one or more stereoisomers are included. The isomers can be separated from the mixture by methods known to those skilled in the art, including: chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of a chiral salt; or preferred isomers can be passed Prepared by asymmetric synthesis.

The invention also includes all suitable isotopic derivatives of the compounds of the invention. An isotope derivative of a compound of the invention is defined as wherein at least one atom is replaced by an atom having the same atomic number but differing in atomic mass from the atomic mass typically found in nature. Examples of isotopes which may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine and chlorine, for example ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, respectively. ¹⁸ F, ³¹ P, ³² P, ³⁵ S and ³⁶ Cl. Some isotopic derivatives of the compounds of the invention, such as those in which a radioisotope such as ³ H or ¹⁴ C is incorporated, are useful in drug and/or substrate tissue distribution studies. Deuterated (i.e., ³ H) and carbon-14 (i.e., ¹⁴ C) isotopes are particularly preferred for their ease of preparation and detectability. Furthermore, substitution with isotopes (e.g., hydrazine, i.e., ² H) can provide some therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and thus may be preferred in some circumstances. . Isotopic derivatives of the compounds of the invention can generally be prepared by conventional procedures, for example by descriptive methods or by the preparations described in the Examples below, using appropriate isotopic derivatives of the appropriate reagents.

The compound of the present invention or a pharmaceutically acceptable salt thereof may be in an amorphous or crystalline form. Furthermore, the compounds of the invention may exist in one or more crystalline forms. Accordingly, the invention includes within its scope all amorphous or crystalline forms of the compounds of the invention.

Those skilled in the art will appreciate that many organic compounds can form complexes with solvents in which they react or precipitate or crystallize from the solvent. These complexes are referred to as "solvates." When the solvent is water, the complex is referred to as a "hydrate." The present invention encompasses all solvates of the compounds of the invention.

In addition, prodrugs are also included within the context of the present invention. The term "prodrug" as used herein refers to a compound which is converted in vivo to an active form thereof having a medical effect by, for example, hydrolysis in blood. Pharmaceutically acceptable prodrugs are described in T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, ACSSymposium Series Vol. 14, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, and D. Fleisher, S. Ramon, and H. Barbra "Improved oral drug delivery: Solubility limitations overcome by the use of prodrugs", Advanced Drug Delivery Reviews (1996) 19(2) 115-130, each This article is incorporated herein by reference.

A prodrug is any covalently bonded carrier which, when administered to a patient, releases the compound of formula (I) in vivo. Prodrugs are typically prepared by modifying functional groups in such a way that the modifications can be cleaved by routine manipulation or in vivo to yield the parent compound. Prodrugs include, for example, compounds of the invention wherein a hydroxy, amine or sulfhydryl group is bonded to any group which, when administered to a patient, can be cleaved to form a hydroxy, amine or sulfhydryl group. Thus, representative examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol, mercapto and amine functional groups of the compounds of formula (I). Further, in the case of a carboxylic acid (-COOH), an ester such as a methyl ester, an ethyl ester or the like can be used. The ester itself may be active and/or may hydrolyze under conditions in humans. Suitable pharmaceutically acceptable in vivo hydrolysable ester groups include those groups which readily decompose in the human body to release the parent acid or a salt thereof.

Pharmaceutical compositions, formulations and kits

In another aspect, the invention provides a pharmaceutical composition comprising a compound of the invention (also referred to as "active ingredient") and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises an effective amount of the active component. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the active component. In some embodiments, the pharmaceutical composition comprises a prophylactically effective amount of the active component.

A pharmaceutically acceptable excipient for use in the present invention refers to a non-toxic carrier, adjuvant or vehicle which does not destroy the pharmacological activity of the compound formulated together. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (eg, human serum white) Protein), buffer substances (such as phosphate), glycine, sorbic acid, potassium sorbate, a mixture of partial glycerides of saturated plant fatty acids, water, salt or electrolyte (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate , sodium chloride, zinc salt, silica gel, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene-polyoxypropylene - Block polymers, polyethylene glycol and lanolin.

The invention also includes kits (e.g., pharmaceutical packs). Kits provided may include a compound of the invention, other therapeutic agents, and first and second containers (eg, vials, ampoules, bottles, syringes, and/or dispersible packages or other materials containing the compounds of the invention, other therapeutic agents) Suitable container). In some embodiments, provided kits can also optionally include a third container containing a pharmaceutically acceptable excipient for diluting or suspending a compound of the invention and/or other therapeutic agent. In some embodiments, a compound of the invention provided in a first container and a second container is combined with other therapeutic agents to form a unit dosage form.

The following formulation examples illustrate representative pharmaceutical compositions that can be prepared in accordance with the present invention. However, the invention is not limited to the following pharmaceutical compositions.

Exemplary Formulation 1 - Tablet: The compound of the present invention in dry powder form can be mixed with the dried gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate was added as a lubricant. The mixture is shaped into 0.3-30 mg tablets (each tablet contains 0.1-10 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 2 - Tablet: The compound of the present invention in dry powder form can be mixed with the dried gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate was added as a lubricant. The mixture is formed into a 30-90 mg tablet (each tablet contains 10-30 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 3 - Tablet: The compound of the invention in dry powder form can be combined with the dried gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate was added as a lubricant. The mixture is shaped into 90-150 mg tablets (30-50 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 4-Tablet: The compound of the invention in dry powder form can be combined with the dried gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate was added as a lubricant. The mixture is formed into a 150-240 mg tablet (each tablet contains 50-80 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 5 - Tablet: The compound of the invention in dry powder form can be combined with the dried gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate was added as a lubricant. The mixture is shaped into 240-270 mg tablets (each tablet contains 80-90 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 6-Tablet: The compound of the invention in dry powder form can be combined with the dried gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate was added as a lubricant. The mixture is shaped into a 270-450 mg tablet (each tablet contains 90-150 mg of active compound) in a tablet press.

Exemplary Formulation 7-Tablet: The compound of the invention in dry powder form can be combined with the dried gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate was added as a lubricant. The mixture is shaped into 450-900 mg tablets (each tablet contains 150-300 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 8-Capsule: The compound of the present invention in dry powder form can be mixed with a starch diluent in a weight ratio of about 1:1. The mixture was filled into 250 mg capsules (each capsule containing 125 mg of active compound).

Exemplary Formulation 9-Liquid: The compound of the present invention (125 mg) can be mixed with sucrose (1.75 g) and xanthan gum (4 mg), and the resulting mixture can be blended, passed through a No. 10 mesh U.S. sieve, and then With previously prepared microcrystalline cellulose and sodium carboxymethylcellulose (11:89, 50 mg) of the aqueous solution was mixed. Sodium benzoate (10 mg), flavor and color are diluted with water and added with stirring. Then, sufficient water can be added to give a total volume of 5 mL.

Exemplary Formulation 10 - Injection: The compound of the invention may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of about 5 mg/mL.

Administration

The pharmaceutical composition provided by the present invention can be administered by a variety of routes including, but not limited to, oral administration, parenteral administration, inhalation administration, topical administration, rectal administration, nasal administration, oral administration, vaginal administration. Drug, administration by implant or other means of administration. For example, parenteral administration as used herein includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, intra-articular administration, intra-arterial administration, intrasynovial administration, intrasternal administration. , intracerebroventricular administration, intralesional administration, and intracranial injection or infusion techniques.

Generally, an effective amount of a compound provided herein is administered. Depending on the circumstances, including the condition being treated, the route of administration selected, the compound actually administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc., the amount of compound actually administered can be determined by the physician. .

When used to prevent a condition of the invention, the compound provided herein is administered to a subject at risk of developing the condition, typically based on a physician's recommendation and administered under the supervision of a physician, at the dosage level as described above. Subjects at risk of developing a particular condition typically include subjects with a family history of the condition, or those subjects that are particularly susceptible to developing the condition by genetic testing or screening.

The pharmaceutical compositions provided herein ("long-term administration") can also be administered chronically. Long-term administration refers to administration of a compound or a pharmaceutical composition thereof for a long period of time, for example, 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or can be continuously administered indefinitely, For example, the rest of the subject. In some embodiments, chronic administration is intended to provide a constant level of the compound in the blood over a prolonged period of time, for example, within a therapeutic window.

The pharmaceutical composition of the present invention can be further delivered using various methods of administration. For example, in some embodiments, a pharmaceutical composition can be administered by bolus injection, for example, to increase the concentration of the compound in the blood to an effective level. The bolus dose depends on the target systemic level of the active ingredient through the body, for example, an intramuscular or subcutaneous bolus dose that causes a slow release of the active ingredient, while a bolus that is delivered directly to the vein (eg, via IV IV drip) ) can be delivered more quickly, so that the concentration of the active ingredient in the blood is rapidly increased to an effective level. In other embodiments, the pharmaceutical composition can be administered in a continuous infusion form, for example, by IV intravenous drip to provide a steady state concentration of the active ingredient in the subject's body. Moreover, in other embodiments, a bolus dose of the pharmaceutical composition can be administered first, followed by continued infusion.

Oral compositions can be in the form of a bulk liquid solution or suspension or bulk powder. More generally, however, the composition is provided in unit dosage form for ease of precise dosing. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human patients and other mammals, each unit containing a predetermined quantity of active ingredient suitable to produce the desired therapeutic effect with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, pre-measured ampoules or syringes of liquid compositions, or in solids Pills, tablets, capsules and the like in the case of a body composition. In such compositions, the compound will generally be a minor component (about 0.1 to about 50% by weight, or preferably about 1 to about 40% by weight), with the remainder being useful for forming the desired form of administration. A carrier or excipient and a processing aid.

For oral doses, a representative regimen is one to five oral doses per day, especially two to four oral doses, typically three oral doses. Using these dosing modes, each dose provides from about 0.01 to about 20 mg/kg of a compound of the invention, each preferably providing from about 0.1 to about 10 mg/kg, especially from about 1 to about 5 mg/kg.

In order to provide a blood level similar to the use of an injectable dose, or a lower blood level than the injectable dose, a transdermal dose is generally selected in an amount of from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably about 0.1. To about 10% by weight, and more preferably from about 0.5 to about 15% by weight.

The injection dose level ranges from about 1 mg/kg/hr to at least 10 mg/kg/hr from about 1 to about 120 hours, especially 24 to 96 hours. In order to obtain a sufficient level of steady state, a preload bolus of about 0.1 mg/kg to about 10 mg/kg or more can also be administered. For a human patient of 40 to 80 kg, the maximum total dose cannot exceed about 2 g/day.

Liquid forms suitable for oral administration may include suitable aqueous or nonaqueous vehicles as well as buffers, suspending and dispersing agents, coloring agents, flavoring agents, and the like. The solid form may include, for example, any of the following components, or a compound having similar properties: a binder, for example, microcrystalline cellulose, tragacanth or gelatin; an excipient such as starch or lactose, a disintegrant, For example, alginic acid, Primogel or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silica; a sweetener such as sucrose or saccharin; or a flavoring agent such as mint, water Methyl salicylate or orange flavoring.

Injectable compositions are typically based on injectable sterile saline or phosphate buffered saline, or other injectable excipients known in the art. As previously mentioned, in such compositions, the active compound will typically be a minor component, often from about 0.05 to 10% by weight, with the remainder being injectable excipients and the like.

The transdermal compositions are typically formulated as topical ointments or creams containing the active ingredient. When formulated as an ointment, the active component is typically combined with a paraffin or water miscible ointment base. Alternatively, the active ingredient can be formulated as a cream with, for example, an oil-in-water cream base. Such transdermal formulations are well known in the art and generally include other ingredients for enhancing stable skin penetration of the active ingredient or formulation. All such known transdermal formulations and components are included within the scope of the invention.

The compounds of the invention may also be administered by transdermal means. Thus, transdermal administration can be accomplished using a reservoir or a porous membrane type, or a patch of a plurality of solid matrices.

The above components of the composition for oral administration, injection or topical administration are merely representative. Other materials and processing techniques, etc., are set forth in Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, part 8 of which is incorporated herein by reference.

The compounds of the invention may also be administered in sustained release form or from a sustained release delivery system. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

The invention further relates to pharmaceutically acceptable formulations of the compounds of the invention. In one embodiment, the formulation comprises water. In another In one embodiment, the formulation comprises a cyclodextrin derivative. The most common cyclodextrins are alpha-, beta- and gamma-cyclodextrins consisting of 6, 7 and 8 alpha-1,4-linked glucose units, respectively, optionally including one on the attached sugar moiety. Or a plurality of substituents including, but not limited to, methylated, hydroxyalkylated, acylated, and sulfoalkyl ether substituted. In some embodiments, the cyclodextrin is a sulfoalkyl ether beta-cyclodextrin, eg, sulfobutylether beta-cyclodextrin, also known as Captisol. See, for example, U.S. 5,376,645. In some embodiments, the formulation comprises hexapropyl-β-cyclodextrin (eg, 10-50% in water).

combination

The compounds of the invention or compositions thereof may be administered in combination with other therapeutic agents to treat the disease. Examples of known therapeutic agents include, but are not limited to, Adriamycin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan ( Topotecan), taxol, interferon, platinum derivatives, taxanes (eg, paclitaxel), vinca alkaloids (eg, vinblastine), anthracycline ( Anthracycline) (eg, doxorubicin), epipodophyllotoxin (eg, etoposide), cisplatin, mTOR inhibitor (eg, rapamycin), Methotrexate, actinomycin D, dolastatin 10, colchicine, emetine, trimetrexate, chlorobenzene Metoprine, cyclosporine, daunorubicin, teniposide, amphotericin, alkylating agent (eg chlorambucil) ), 5-fluorouracil, camptothecin, cisplatin, metronidazole (metro) nidazole) and Gleevec ^TM. In other embodiments, the compounds of the invention are administered in combination with a biological agent such as Avastin or VECTIBIX.

In some embodiments, a compound of the invention or a composition thereof can be administered in combination with any one or more anti-proliferative or chemotherapeutic agents selected from the group consisting of: abarelix, aldileukin (aldesleukin), alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, day Asparaginase, azacitidine, BCG Live, bevacuzimab, fluorouracil, bexarotene, bleomycin, bortezomib, white Busulfan, calbuterone, capecitabine, camptothecin, carboplatin, carmustine, celecoxib, cetuximab Antibiotic (cetuximab), chlorambucil, cladribine, clofarabine, cyclophosphamide, cytarabine, actinomycin D, darbepoetin ( (darbepoetin alfa), daunorubicin, denilukin , dexrazoxane, docetaxel, doxorubicin, doxorubicin hydrochloride, dromostanolone propionate, epirubicin, ebertin alpha Epoetin alfa), erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, filgrastim, fluururidine ), fludarabine, fulvestrant, gefitinib, gemcitabine, gemtuzumab, goserelin acetate, acetic acid ammonia Histrelin acetate, hydroxyurea, ibritumomab, idarubicin, ifosfamide, Imatinib mesylate, interferon alpha-2a, interferon alpha-2b, irinotecan, lenalidomide, letrozole, formyltetrahydrogen Leucovorin, leuprolide acetate, levamisole, lomustine, megestrol acetate, melphalan, mercaptopurine ), 6-MP, sodium mesaconate (mesna), methotrexate, methoxsalen, mitomycin C, mitotan, mitoxantrone Mitoxrolone, nandrolone, nelarabine, nofetumomab, oprelvekin, oxaliplatin, paclitaxel, palifermin, Pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, pentastatin (pemetreronate) Pentostatin), pipobroman, plicamycin, porfimer sodium, Procarbazine, quinacrine, rasburicase, rituximab, sargramostim, sorafenib, streptozotocin Streptozocin, sunitinib maleate, talc, tamoxifen, temozolomide, teniposide, VM-26, testolactone, Thioguanine, 6-TG, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, dimension Formic acid (tretinoin), ATRA, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, zoledronate or Zoledronic acid.

Other examples of therapeutic agents to which the compounds of the invention may also be combined include, but are not limited to, therapeutic agents for Alzheimer's Disease, such as donepezil hydrochloride and rivastigmine. ; therapeutic agents for Parkinson's Disease, such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole ( Pramipexole), bromocriptine, pergolide, trihexephendyl, and amantadine; a therapeutic agent for multiple sclerosis (MS), Such as beta interferon, glatiramer acetate and mitoxantrone; therapeutic agents for asthma, such as albuterol and montelukast; therapeutic agents for schizophrenia, such as Zyprexa, risperdal, seroquel, and haloperidol; anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1RA, azathioprine , cyclophosphamide and sulfasalazine (sulfasalazi) Ne); immunomodulators and immunosuppressive agents, such as cyclosporin, tacrolimus, rapamycin, mycophenolate mofetil, interferon, corticosteroids, Cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, riluzole, and anti-par a drug for the treatment of cardiovascular diseases, such as a beta blocker, an ACE inhibitor, a diuretic, a nitrate, a calcium channel blocker, and a statin; a therapeutic agent for liver diseases, Such as corticosteroids, cholestyramine, interferon and antiviral agents; therapeutic agents for blood disorders such as corticosteroids, anti-leukemia agents and growth factors; and therapeutic agents for immunodeficiency disorders, such as gamma spheres protein.

Those other active agents can be administered separately from the compositions containing the compounds of the invention as part of a multiple dosing regimen. Alternatively, those active agents may be part of a single dosage form, mixed with a compound of the invention in a single composition. If administered as multiple doses If a portion of the regimen is administered, then the two active agents can be provided simultaneously, sequentially, or at intervals from one another (usually within 5 hours of each other).

treatment

The invention provides a method for inhibiting a protein tyrosine kinase (such as EGFR kinase) or treating a disease (such as cancer, cell proliferative disease, inflammation, infection, immune disease, organ transplantation, viral disease, cardiovascular disease or A method of metabolic disease comprising the steps of administering a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug thereof, to a subject in need of treatment Or an isotope derivative, or a pharmaceutical composition of the invention.

The compounds of the invention are useful in the treatment of cancer caused by EGFR. In particular, the compounds are useful for treating cancer caused by EGFR expressing an EGFR mutant and for treating cancer caused by EGFR that is refractory to RTKI therapy (eg, erlotinib or gefitinib).

The compounds of the invention are inhibitors of at least one mutant of EGFR and are therefore suitable for treatment with one or more EGFR mutants (eg, deletion mutations, activating mutations, resistance mutations, or combinations thereof, specific examples include T790M mutations, One or more disorders associated with the activity of the L858R mutation and the L858R/T790M double mutation. Accordingly, in a particular embodiment, the invention provides a method of treating a mutant EGFR mediated disorder comprising administering a compound of the invention, or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, to a patient in need thereof a solvate, hydrate, crystal form, prodrug or isotope derivative, or a step of administering a pharmaceutical composition of the invention.

Cancers treatable by the compounds of the invention include, but are not limited to, non-small cell lung cancer (NSCLS), small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, Hyperproliferative diseases such as gastrointestinal stromal tumors, leukemia, histiocytic lymphoma, and nasopharyngeal carcinoma. In addition, the compounds of the invention may also be used to maintain the maintenance of cancer recurrence in patients in need of such treatment.

An effective amount of a compound of the invention will generally be administered in a single or multiple doses at an average daily dose of from 0.01 mg to 50 mg of compound per kilogram of patient body weight, preferably from 0.1 mg to 25 mg of compound per kilogram of patient body weight. In general, the compounds of the invention may be administered to a patient in need of such treatment in a daily dosage range of from about 1 mg to about 3500 mg per patient, preferably from 10 mg to 1000 mg. For example, the daily dose per patient can be 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 500, 600, 700, 800, 900 or 1000 mg. It can be administered one or more times daily, weekly (or several days apart) or on an intermittent schedule. For example, the compound can be administered one or more times per day on a weekly basis (e.g., every Monday), continually or for several weeks, such as 4-10 weeks. Alternatively, the administration may be continued for several days (e.g., 2-10 days), followed by a few days (e.g., 1-30 days) without administration of the compound, and the cycle may be repeated indefinitely or repeated for a given number of times, such as 4-10 Cycles. For example, the compounds of the invention may be administered daily for 5 days, then intermittently for 9 days, then administered daily for 5 days, then intermittent for 9 days, and so on, optionally repeating the cycle or repeating 4-10 times.

When EGFR-TKI (e.g., erlotinib or gefitinib) is used in combination with a compound of the invention, the individual components of the combination therapy can be administered at their dosage level and regimen for monotherapy. For example, erlotinib has been orally administered at a dose of 150 mg per day for the treatment of non-small cell lung cancer, and has been orally administered at a dose of 100 mg per day for pancreatic cancer. In another example, gefitinib is used to treat non- Small cell lung cancer has been administered orally at 250 mg per day.

Preferably, when EGFR-TKI (e.g., erlotinib or gefitinib) is used in combination with a compound of the invention, the dosage level of one or both components is reduced compared to when used alone.

Example

The following examples are provided to provide those skilled in the art with a complete disclosure and description of how to make, prepare, and evaluate the methods and compounds claimed herein.

resolve resolution

The compounds of the present invention can be prepared according to conventional methods in the art and using suitable reagents, starting materials, and purification methods known to those skilled in the art.

The preparation of the structural compound of the formula I of the present invention is more specifically described below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention may also be conveniently prepared by combining various synthetic methods described in the specification or known in the art, and such combinations are readily made by those skilled in the art to which the present invention pertains.

Usually, in the preparation, each reaction is usually carried out in an inert solvent at room temperature to reflux temperature (e.g., 0 ° C to 100 ° C, preferably 0 ° C to 80 ° C). The reaction time is usually from 0.1 to 60 hours, preferably from 0.5 to 24 hours.

Example 1 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1-H-indol-2-yl)thieno[3 ,2-d]pyrimidine-2- Amino}phenyl)prop-2-enamide (Compound T-1)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 3.

Under a nitrogen atmosphere, 2,4-dichlorothieno[3,2-d]pyrimidine (Compound 2, 393 mg, 1.9 mmol), Na ₂ CO ₃ (201 mg, 1.9 mmol), Pd(PPh ₃ ) ₄ ( 44 mg, 0.038 mmol) was added to a solution of 1-tert-butoxycarbonyl-1H-indole-2-boronic acid (0.5 g, 1.9 mmol) in ethanol (7 mL), and the mixture was stirred at 70 ° C for 2 hr then cooled The solvent was removed to room temperature and purified by column chromatography to yield a white solid product (yield: 96%). LC-MS (APCI): m / z = 386.1 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 8.28-8.24 (m, 1H), 8.13 (d, J = 5.7Hz , 1H), 7.70-7.67 (m, 1H), 7.58 (d, J = 5.7 Hz, 1H), 7.51-7.45 (m, 1H), 7.37-7.31 (m, 1H), 7.25 (s, 1H), 1.32 (s, 9H).

Step 2 Synthesis of Compound 5.

Compound 4 (186 mg, 1.2 mmol) and p-toluenesulfonic acid monohydrate (228 mg, 1.44 mmol) were added to a solution of compound 3 (385 mg, 1.2 mmol) in 2-pentanol (6 mL) and heated to 105 ° C. After reacting overnight, the reaction mixture was cooled to room temperature. LC-MS (APCI): m / z = 436.1 (M + 1) +.

Step 3 Synthesis of Compound 7.

Compound 6 (280 mg, 2.75 mmol) and potassium carbonate (507 mg, 3.68 mmol) were added to a solution of compound 5 (400 mg, 0.92 mmol) in N,N-dimethylformamide (DMF, 5 mL) After stirring at room temperature for 3 hours, it was filtered, and extracted with dichloromethane. LC-MS (APCI): m / z = 518.1 (M + 1) +, 1 H NMR (400MHz, CDCl 3) (δ / ppm) 9.94 (br s, 1H), 9.59 (s, 1H), 7.94 ( d, J = 5.6 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.60 (s, 1H), 7.46 (d, J = 1.6 Hz, 1H), 7.38-7.33 (m, 2H), 7.18 (t, J = 15.2 Hz, 1H), 6.67 (s, 1H), 3.99 (s, 3H), 3.32 (t, J = 6.8 Hz, 2H), 2.94 (s, 3H), 2.61 (t, J = 6.8 Hz, 2H), 2.29 (s, 6H).

Step 4 Synthesis of Compound 8.

Iron powder (276 mg, 4.9 mmol) and ammonium chloride (66 mg, 1.23 mmol) were added to a solution of compound 7 (426 mg, 0.82 mmol) in ethanol (6 mL) and water (2 mL). ℃ after reaction for 2 hours, cooled to room temperature, the solvent was removed under reduced pressure and filtered with _{CH 3 Cl: i-PrOH (} 3: 1) extraction, the organic phase was collected to give a brown solid product was used directly in the next step.

Step 5 Synthesis of Compound T-1.

The reaction product of the previous step was dissolved in dichloromethane (25 mL), triethylamine (0.1 g, 1 mmol) was added, cooled to -20 ° C, and a solution of acryloyl chloride (compound 9, 81 mg, 0.9 mmol) in dichloromethane was added. The reaction was carried out at this temperature for 2 hours, and the reaction was quenched with water, and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to yield white solid product (yield: 45%). LC-MS (APCI): m / z = 542.2 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 11.61 (br s, 1H), 10.45 (br s, 1H), 10.01 (s, 1H), 7.93-7.90 (m, 2H), 7.82 (s, 1H), 7.75 (d, J = 8.1 Hz, 1H), 7.53 (d, J = 1.5 Hz, 1H), 7.34 - 7.28 ( m, 2H), 7.12 (t, J = 8.1 Hz, 1H), 6.83 (s, 1H), 6.70-6.64 (m, 1H), 5.51-6.48 (m, 1H), 5.86-5.82 (m, 1H) , 3.91 (s, 3H), 2.92 (t, J = 5.4 Hz, 2H), 2.75 (s, 3H), 2.31-2.29 (m, 8H).

Example 2 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1-methylindol-3-yl)thieno[3 ,2-d]pyrimidine -2-yl]amino}phenyl)prop-2-enamide (Compound T-2)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 10.

To a continuously stirred 3-neck round bottom flask was added 2,4-dichlorothieno[3,2-d]pyrimidine (compound 2, 1.0 g, 4.88 mmol) and 1,2-dichloroethane under a nitrogen atmosphere. (13 mL), after heating to 60 ° C, FeCl ₃ (0.78 g, 4.81 mmol) was added, and 1-methyl hydrazine (0.72 g, 5.49 mmol) of 1,2-dichloroethane (3 mL) was added dropwise to the reaction system. After the completion of the dropwise addition, the reaction was carried out at 60 ° C overnight, diluted with 20 mL of methanol, filtered, and the organic phase was collected, and the mixture was stirred for 20 minutes, washed with water, ethyl acetate, dichloromethane, and dried The yield was 65%. LC-MS (APCI): m / z = 300 (M + 1) +, 1 H NMR (300MHz, DMSO-d 6) (δ / ppm) 8.64 (d, J = 4.8Hz, 1H), 8.57 (d , J = 4.8 Hz, 1H), 8.48 (s, 1H), 7.62-7.60 (m, 2H), 7.36-7.32 (m, 2H), 4.00 (s, 3H).

Step 2 Synthesis of Compound 11.

Compound 4 (186 mg, 1.2 mmol) and p-toluenesulfonic acid monohydrate (205 mg, 1.2 mmol) were added to a solution of compound 10 (300 mg, 1.0 mmol) in 2-pentanol (6 mL) and heated to 105 ° C. After reacting overnight, the reaction mixture was cooled to room temperature. LC-MS (APCI): m / z = 450 (M + 1) +.

Step 3 Synthesis of Compound 12.

Compound 6 (102 mg, 1.0 mmol) and potassium carbonate (507 mg, 3.68 mmol) were added to a solution of compound 11 (385 mg, 0.78 mmol) in N,N-dimethylformamide (10 mL) After stirring for 3 hours, it was filtered, and the filter cake was dissolved in water, filtered, washed with water, dried and purified by column chromatography to yield 371 mg of product as a yellow solid. LC-MS (APCI): m / z = 532 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 9.36 (s, 1H), 8.66 (d, J = 7.5Hz, 1H ), 8.00 (s, 1H), 7.83 (d, J = 5.4 Hz, 1H), 7.62 (s, 1H), 7.40-7.34 (s, 4H), 6.70 (s, 1H), 4.01 (s, 3H) , 3.94 (s, 3H), 3.27 (t, J = 6.9 Hz, 2H), 2.90 (s, 3H), 2.57 (t, J = 6.9 Hz, 2H), 2.27 (s, 6H).

Step 4 Synthesis of Compound 13.

Iron powder (235 mg, 4.2 mmol) and ammonium chloride (37 mg, 0.7 mmol) were added to a solution of compound 12 (371 mg, 0.7 mmol) in ethanol (10 mL) and water (2 mL). After reacting for 2 hours at ° C, the mixture was cooled to room temperature, filtered, evaporated, evaporated, evaporated, evaporated. LC-MS (APCI): m / z = 502 (M + 1) +.

Step 5 Synthesis of Compound T-2.

Compound 13 (350 mg, 0.7 mmol), tetrahydrofuran (10 mL), water (1 mL) was added to a continuously stirred three-necked round bottom flask. The reaction mixture was cooled to 0 ° C and compound 14 (107 mg, 0.84 mmol) was added. After stirring for 15 minutes at room temperature, sodium hydroxide (112 mg, 2.8 mmol) was added, and the mixture was heated to 65 ° C and reacted for 15 hours, extracted with water and ethyl acetate, washed with water, dried and purified by column chromatography The yield was 41%. LC-MS (APCI): m / z = 556 (M + 1) +, 1 H NMR (300MHz, DMSO-d 6) (δ / ppm) 10.09 (s, 1H), 8.82 (s, 1H), 8.50 (d, J = 8.1 Hz, 1H), 8.31 (s, 1H), 8.26 (d, J = 5.4 Hz, 1H), 8.18 (s, 1H), 7.53 (d, J = 8.1 Hz, 1H), 7.23 -7.30 (m, 2H), 7.11 (t, J = 6.9 Hz, 1H), 7.04 (s, 1H), 6.36-6.42 (m, 1H), 6.19 (dd, J = 17.1 Hz, 2.4 Hz, 1H) , 5.70-5.75 (m, 1H), 3.97 (s, 3H), 3.80 (s, 3H), 2.92 (s, 2H), 2.73 (s, 3H), 2.37 (s, 2H), 2.23 (s, 6H) ).

Example 3 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(pyrazolo[1,5-a]pyridin-3-yl) )thieno[3,2-d] Pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-3)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 16.

2,4-Dichlorothieno[3,2-d]pyrimidine (Compound 2, 1.5 g, 7.32 mmol), vinyl n-butyl ether (Compound 15, 2.2 g, 21.95 mmol) and triethylamine (777 mg) , 7.68 mmol) was dissolved in 20 mL of polyethylene glycol (PEG200). After adding Pd(OAc) ₂ (82 mg, 366 μmol), the reaction solution was heated to 80 ° C, and reacted at this temperature for 16 hours, and poured into water. After extraction with ethyl acetate, the organic phase was collected and used directly in the next step.

Step 2 Synthesis of Compound 18.

Compound 16 (1.9 g, 7.07 mmol), 1-aminopyridine iodide (Compound 17, 1.57 g, 7.07 mmol) and potassium carbonate (2.44 g, 17.67 mmol) were dissolved in 10 mL of DMF solution and heated to 110 ° C. After an hour, it was cooled to room temperature, filtered, and the filter cake was washed with water and evaporated to give a white solid product (yield: 20%). LC-MS (APCI): m / z = 287.3 (M + 1) +, 1 HNMR (300MHz, CDCl 3) (δ / ppm) 8.89 (d, 1H, J = 9Hz), 8.79 (s, 1H), 8.63 (d, 1H, J = 6.9 Hz), 8.04 (d, 1H, J = 4.5 Hz), 7.58 - 7.55 (m, 2H), 7.12 - 7.06 (m, 1H).

Step 3 Synthesis of Compound 19.

Compound 18 (400 mg, 1.4 mmol) and compound 4 (1.04 g, 5.58 mmol) in 3 mL of trifluoroacetic acid were heated to 150 ° C and reacted for 4 hours, cooled to room temperature and filtered, and the filtrate was collected and washed with saturated sodium hydrogen carbonate solution. After drying, the product was obtained as a yellow solid, 330 mg, yield 50%. LC-MS (APCI): m / z = 437.4 (M + 1) +.

Step 4 Synthesis of Compound 20.

After stirring a solution of compound 6 (140 mg, 1.37 mmol), potassium carbonate (285 mg, 2.06 mmol) and compound 19 (300 mg, 0.69 mmol) in N, N-dimethylformamide (4 mL), After extracting the organic phase by water and dichloromethane, the crude brown solid was purified by thin layer chromatography (yield: 66%). LC-MS (APCI): m / z = 519.3 (M + 1) +.

Step 5 Synthesis of Compound T-3.

Heat a solution of iron powder (35 mg, 0.64 mmol), ammonium chloride (41 mg, 0.7 mmol) and compound 20 (66 mg, 0.1 mmol) in ethanol (6 mL) and water (3 mL) to 90 ° C under a nitrogen atmosphere and react 2 After a few hours, it was cooled to room temperature, and the solvent was evaporated under reduced pressure. The yellow solid and triethylamine (23 mg, 225 μmol) were dissolved in 10 mL of dichloromethane, cooled to -15 ° C, then acryloyl chloride (10 mg, 112 μmol) was added, stirring was continued at -10 ° C for 15 minutes, and 10 mL of NaHCO was poured. _The solution was extracted with dichloromethane, and the organic phase was collected. LC-MS (APCI): m / z = 543.4 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 10.06 (s, 1H), 9.67 (s, 1H), 8.79-8.77 (m, 2H), 5.58-8.56 (m, 1H), 7.86-7.85 (m, 1H), 7.56 (s, 1H), 7.48 (d, J = 4.2 Hz, 1H), 7.40-7.35 (m, 1H) ), 6.97-6.94 (m, 1H), 6.82 (s, 1H), 6.38-6.34 (m, 2H), 2.70-5.67 (m, 1H), 3.91 (s, 3H), 2.94-2.90 (m, 2H) ), 2.70 (s, 3H), 2.38-2.33 (m, 2H), 2.29 (s, 6H).

Example 4 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1H-benzo[d]imidazol-1-yl)thiophene [3,2-d] Pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-4)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 22.

1H-benzo[d]imidazole (0.56 g, 0.5 mmol) and potassium carbonate (0.83 g, 0.6 mmol) were sequentially added to 2,4-dichlorothieno[3,2-d]pyrimidine under a nitrogen atmosphere. A solution of the compound 2, 1.0 g, 0.5 mmol) in DMF (5 mL) was stirred at room temperature overnight, poured into water (80 mL), a white solid was formed, filtered and washed with water to give the desired product (1.1 g). LC-MS (APCI): m / z = 287.0 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 8.86 (s, 1H), 8.49-8.46 (m, 1H), 8.18 (d, J = 6.0 Hz, 1H), 7.92 - 7.89 (m, 1H), 7.66 (d, J = 6.0 Hz, 1H), 7.52 - 7.46 (m, 2H).

Step 2 Synthesis of Compound 23.

Compound 22 (290 mg, 1.0 mmol), Compound 4 (223 mg, 1.2 mmol), p-toluenesulfonic acid (205 mg, 1.2 mmol) and 2-pentanol were added to a continuously stirred 3-neck round bottom flask under nitrogen. It was heated to 105 ° C and allowed to react at this temperature overnight, cooled to room temperature, filtered and dried to give a brown solid product (yield: 94%). LC-MS (APCI): m / z = 437.0 (M + 1) +.

Step 3 Synthesis of Compound 24.

Compound 23 (350 mg, 0.78 mmol), compound 6 (102 mg, 1 mmol), potassium carbonate (138 mg, 1 mmol) and DMF (10 mL) were added to a three-neck round bottom flask, and stirred at room temperature overnight. After 20 mL of water, the mixture was filtered and washed with water, and dried and purified by column chromatography to yield 371 mg of product as a yellow solid. LC-MS (APCI): m / z = 519 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 9.20 (s, 1H), 8.79 (s, 1H), 8.30-8.27 (m, 1H), 7.99 (d, J = 5.4 Hz, 1H), 7.92-7.89 (m, 1H), 7.65 (s, 1H), 7.48-7.43 (m, 3H), 6.71 (s, 1H), 4.01 (s, 3H), 3.30 (t, J = 6.9 Hz, 2H), 2.90 (s, 3H), 2.59 (t, J = 6.9 Hz, 2H), 2.29 (s, 6H).

Step 4 Synthesis of Compound 25.

An iron powder (235 mg, 4.2 mmol), ammonium chloride (37 mg, 0.7 mmol) and a solution of compound 24 (371 mg, 0.7 mmol) in ethanol (10 mL) and water (3 mL) were heated to reflux and react for 2 hours under a nitrogen atmosphere. Thereafter, the mixture was cooled to room temperature, and the solvent was removed by filtration under reduced pressure. LC-MS (APCI): m / z = 489 (M + 1) +.

Step 5 Synthesis of Compound T-4.

Compound 25 (342 mg, 0.7 mmol) was dissolved in 50 mL of dichloromethane under a nitrogen atmosphere, and triethylamine (106 mg, 1.05 mmol) was added dropwise, and after completion of the addition, the reaction system was cooled to -20 ° C. The acryloyl chloride (76 mg, 0.84 mmol) was slowly added at a temperature, stirred for 15 minutes, water was added, and the organic phase was separated and washed with water and a saturated aqueous sodium hydrogen carbonate solution, and the organic phase was collected and purified by column chromatography to yield 156 mg of product as a yellow solid. It is 41%. LC-MS (APCI): m / z = 543 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 9.97 (br s, 1H), 9.72 (s, 1H), 8.88 ( s, 1H), 8.26-8.23 (m, 1H), 7.95 (d, J = 5.4 Hz, 1H), 7.90-7.87 (m, 1H), 7.69 (s, 1H), 7.55 (d, J = 5.4 Hz) , 1H), 7.42-7.36 (m, 2H), 6.81 (s, 1H), 6.44 (d, J = 6.0 Hz, 2H), 5.70 (t, J = 6.0 Hz, 1H), 3.91 (s, 3H) , 2.96 (t, J = 6.0 Hz, 2H), 2.71 (s, 3H), 2.44 (t, J = 6.0 Hz, 2H), 2.35 (s, 6H).

Example 5 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1H-benzo[d]triazol-1-yl) Thio[3,2-d] Pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-5)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 27.

1H-benzo[d]triazole (0.56 g, 0.5 mmol) and potassium carbonate (0.83 g, 0.6 mmol) were sequentially added to 2,4-dichlorothieno[3,2-d] under a nitrogen atmosphere. A solution of the pyrimidine (Compound 2, 1.0 g, 0.5 mmol) in DMF (5 mL) was stirred at 60 ° C overnight, and poured into water (80 mL) to give a white solid, which was filtered and washed with water to give the desired product 1.1 g. LC-MS (APCI): m / z = 288.1 (M + 1) +.

Step 2 Synthesis of Compound 28.

Compound 27 (290 mg, 1.0 mmol), compound 4 (223 mg, 1.2 mmol), p-toluenesulfonic acid (205 mg, 1.2 mmol) and 2-pentanol (6 mL) were added to a continuously stirred three-necked circle under nitrogen. The bottom flask was heated to 105 ° C and allowed to react at this temperature overnight, cooled to room temperature, filtered and dried to give a brown solid product (yield: 94%). LC-MS (APCI): m / z = 438 (M + 1) +.

Step 3 Synthesis of Compound 29.

Compound 28 (350 mg, 0.78 mmol), compound 6 (102 mg, 1 mmol), potassium carbonate (138 mg, 1 mmol) and DMF (10 mL) were added to a three-necked round bottom flask, and stirred at room temperature overnight. After 20 mL of water, the mixture was filtered and washed with water, and dried and purified by column chromatography to yield 280 mg of product as a yellow solid. LC-MS (APCI): m / z = 520 (M + 1) +, 1 H NMR (300MHz, CDCl 3) (δ / ppm) 9.09 (s, 1H), 8.68 (d, J = 8.1Hz, 1H ), 8.18 (d, J = 8.4 Hz, 1H), 8.06 (d, J = 6.0 Hz, 1H), 7.72 - 7.66 (m, 1H), 7.56 - 7.51 (m, 2H), 7.40 (d, J = 5.4 Hz, 1H), 6.72 (s, 1H), 4.02 (s, 3H), 3.32 (t, J = 6.9 Hz, 2H), 2.92 (s, 3H), 2.30 (s, 6H).

Step 4 Synthesis of Compound 30.

Iron powder (170 mg, 3 mmol), ammonium chloride (27 mg, 0.5 mmol) was added to a solution of compound 29 (260 mg, 0.5 mmol) in ethanol (10 mL) and water (3 mL). after cooling to room temperature, the solvent was removed under reduced pressure and was filtered, dried and extracted with dichloromethane after addition to give a yellow solid 245mg, LC-MS (APCI) : m / z = 490.2 (m + 1) +.

Step 5 Synthesis of Compound T-5.

Under a nitrogen atmosphere, compound 30 (245 mg, 0.5 mmol) was dissolved in 100 mL of dichloromethane, and triethylamine (75 mg, 0.7 mmol) was added dropwise, and after completion of the addition, the reaction system was cooled to -20 ° C. The acryloyl chloride (54 mg, 0.6 mmol) was slowly added at a temperature, stirred for 15 minutes, water was added, and the organic phase was separated and washed with water and a saturated sodium hydrogen carbonate solution, and the organic phase was collected and purified by column chromatography to yield 166 mg of product as a yellow solid. It is 61%. LC-MS (APCI): m / z = 544 (M + 1) +, 1 H NMR (300MHz, DMSO-d 6) (δ / ppm) 10.08 (s, 1H), 9.02 (s, 1H), 8.55 (s, 2H), 8.43 (d, J = 5.1 Hz, 1H), 7.63 - 7.58 (m, 2H), 7.36 (d, J = 6.0 Hz, 1H), 7.07 (s, 1H), 6.51 (br s , 1H), 6.21-6.16 (m, 1H), 5.74-5.70 (m, 1H), 3.77 (s, 3H), 3.02-2.97 (m, 2H), 2.76 (s, 3H), 2.55-2.53 (m , 2H), 2.32 (s, 6H).

Example 6 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(5,6-dihydro-4H-pyrrolo[3,2] , 1-ij]quinoline-1- Thio[3,2-d]pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-6)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 32.

Under the protection of nitrogen, ferric chloride (1.5 g, 9 mmol) and 5,6-dihydro-4Hpyrrolo[3,2,1-ij]quinoline (1 g, 6.4 mmol) were sequentially added to 2,4-di A solution of chlorothieno[3,2-d]pyrimidine (Compound 2, 1.13 g, 6.97 mmol) in 1,2-dichloroethane (20 mL), heated to 60 ° C and stirred overnight, then cooled to room temperature and poured 100 mL of water was stirred for 30 minutes, and a white solid was formed, which was filtered and washed with water to give 1.5 g of desired product. LC-MS (APCI): m / z = 326.1 (M + 1) +.

Step 2 Synthesis of Compound 33.

Compound 4 (230 mg, 1.2 mmol), p-toluenesulfonic acid (230 mg, 1.4 mmol) was added to a solution of compound 32 (400 mg, 1.2 mmol) in 2-pentanol (6 mL) and heated to 105 ° C. The reaction was allowed to proceed at this temperature overnight, cooled to room temperature, stirred in a solution of petroleum ether and ethanol (1:1) for 15 minutes, filtered, and dried to give a green solid product 350 mg, yield 61%. LC-MS (APCI): m / z = 476.2 (M + 1) +.

Step 3 Synthesis of Compound 34.

Compound 6 (300 mg, 3 mmol) and potassium carbonate (550 mg, 4 mmol) were added to a solution of compound 33 (475 mg, 1.0 mmol) in DMF (10 mL), and then stirred at room temperature for 3 hr. The filter cake was washed with water, dried and purified by column chromatography to yield 260 mg of product as a red solid, yield 47%. LC-MS (APCI): m / z = 558.2 (M + 1) +, 1 H NMR (400MHz, CDCl 3) (δ / ppm) 9.39 (s, 1H), 8.40 (d, J = 8.0Hz, 1H ), 8.05 (s, 1H), 7.84 (d, J = 5.6 Hz, 1H), 7.64 (s, 1H), 7.40 (d, J = 5.6 Hz, 1H), 7.25 (t, J = 8.0 Hz, 1H) ), 7.07 (d, J = 6.8 Hz, 1H), 6.75 (s, 1H), 4.33 (t, J = 5.6 Hz, 2H), 4.04 (s, 3H), 3.33 (t, J = 7.2 Hz, 2H) ), 3.07 (t, J = 9.6 Hz, 2H), 2.91 (s, 3H), 2.66 (t, J = 7.2 Hz, 2H), 2.36-2.32 (m, 8H).

Step 4 Synthesis of Compound 35.

Iron powder (80 mg, 1.4 mmol) and ammonium chloride (23 mg, 0.44 mmol) were added to a solution of compound 34 (160 mg, 0.29 mmol) in ethanol (3 mL) and water (1 mL). After reacting for 2 hours at ° C, the reaction mixture was cooled to room temperature, and the ethanol was removed under reduced pressure.

Step 5 Synthesis of Compound T-6.

The product obtained in the previous step was dissolved in dichloromethane (15 mL), triethylamine (40 mg, 1.3 eq) was added and cooled to -20 ° C, acryloyl chloride (32 mg, 0.35 mmol) was slowly added at this temperature and stirred. After 1 hour, the reaction was quenched with water, and the organic layer was separated, washed with dichloromethane, and then washed with EtOAc. LC-MS (APCI): m / z = 582.2 (M + 1) +, 1 H NMR (300MHz, CDCl 3 + MeOH-d 4) (δ / ppm) 8.94 (s, 1H), 8.27 (d, J = 4.8 Hz, 1H), 8.06 (s, 1H), 7.87 (d, J = 3.6 Hz, 1H), 7.33 (d, J = 3.0 Hz, 1H), 7.13 (t, J = 4.2 Hz, 1H), 6.98 (d, J = 4.2 Hz, 1H), 6.76 (s, 1H), 6.68-6.63 (m, 1H), 6.37-6.33 (m, 1H), 5.77-4.75 (m, 1H), 4.30 (t, J=3.3 Hz, 2H), 3.95 (s, 3H), 3.35-3.33 (m, 2H), 3.17-3.14 (m, 2H), 3.02 (t, J = 3.3 Hz, 2H), 2.77 (s, 6H) ), 2.68 (s, 3H), 2.30-2.28 (m, 2H).

Example 7 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1H-indazol-1-yl)thieno[3,2 -d]pyrimidine-2- Amino}phenyl)prop-2-enamide (Compound T-7)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 37.

Compound 36 (2.44 g, 10 mmol), Compound 2 (2.05 g, 10 mmol), potassium carbonate (1.66 g, 12 mmol) and DMF (30 mL) were sequentially added to a continuously stirred single-necked round bottom flask under nitrogen. After the completion of the addition, the mixture was stirred at room temperature overnight, and 20 mL of water was added to the reaction mixture. After filtration, the cake was washed with water and dried to give a white solid product (3.2 g, yield: 78%). LC-MS (APCI): m / z = 413 (M + 1) +.

Step 2 Synthesis of Compound 38.

Compound 37 (2.06 g, 5 mmol), THF (50 mL), tetramethylethylenediamine (2 g, 17 mmol), Pd(dppf)Cl ₂ were sequentially added to a continuously stirred single-necked round bottom flask under a nitrogen atmosphere. (204.2 g, 0.25 mmol) and NaBH ₄ (0.65 g, 17 mmol) were stirred at room temperature for 2 hr, and water (20 mL) was added. After filtration, the cake was washed with water and dried to give a white solid. LC-MS (APCI): m / z = 287 (M + 1) +.

Step 3 Synthesis of Compound 39.

Compound 38 (900 mg, 3.15 mmol), compound 4 (700 mg, 3.78 mmol), p-toluenesulfonic acid (651 mg, 3.78 mmol) and 2-pentanol (6 mL) were added to a single-neck round under nitrogen. After reacting for 3 hours at 105 ° C in a bottom flask, the cake was filtered and washed with 2-pentanol and water, and dried to give a brown solid product (1 g, yield: 73%). LC-MS (APCI): m / z = 437 (M + 1) +.

Step 4 Synthesis of Compound 40.

Compound 39 (436 mg, 1 mmol), Compound 6 (120 mg, 1.2 mmol), potassium carbonate (200 mg, 1.5 mmol) and DMF (10 mL) were added to a three-necked round bottom flask and stirred at room temperature under nitrogen. After an hour, it was diluted with 20 mL of water, filtered, washed with water, and the cake was dissolved in dichloromethane, and then extracted, and the organic phase was collected and purified by column chromatography to give a white solid product (yield: 77%). LC-MS (APCI): m / z = 519 (M + 1) +. ¹ H NMR (300 MHz, CDCl ₃ ) (δ / ppm) 9.14 (s, 1H), 8.83 (d, J = 8.4 Hz, 1H), 8.30 (s, 1H), 7.94 (d, J = 5.7 Hz, 1H) ), 7.78 (d, J = 7.2 Hz, 1H), 7.59 - 7.53 (m, 1H), 7.45 (s, 1H), 7.32 - 7.37 (m, 2H), 6.97 (s, 1H), 4.00 (s, 3H), 3.30 (t, J = 6.9 Hz, 2H), 2.91 (s, 3H), 2.59 (t, J = 7.5 Hz, 2H), 2.29 (s, 6H).

Step 5 Synthesis of Compound 41.

Compound 40 (371 mg, 0.7 mmol), iron powder (235 mg, 4.2 mmol) and ammonium chloride (37 mg, 0.7 mmol) were added to a mixture of water/ethanol (10 mL / 2 mL) in a mixture. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed and water was added, and extracted with dichloromethane, and the organic phase was collected to give a product as a yellow solid (230 mg, yield: 72%). LC-MS (APCI): m / z = 489 (M + 1) +.

Step 6 Synthesis of Compound T-7.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 50 mL of dichloromethane was added sequentially to compound 41 (245 mg, 0.5 mmol). After it was completely dissolved, triethylamine (75 mg, 0.75 mmol) was added dropwise. The system was cooled to -20 ° C, and the compound 9 (54 mg, 0.6 mmol) was slowly added, and the stirring was continued for 15 minutes. The reaction was quenched with water and the organic phase was separated and washed with water and saturated sodium hydrogen carbonate solution. Chromatography gave 156 mg of product as a yellow solid. LC-MS (APCI): m / z = 543 (M + 1) +. ^{1 H NMR (300MHz, DMSO-} d 6) (δ / ppm) 10.12 (s, 1H), 9.67 (s, 1H), 8.91 (d, J = 8.4Hz, 1H), 8.33 (s, 1H), 7.93 (d, J = 5.7 Hz, 1H), 7.79 (d, J = 7.8 Hz, 1H), 7.45 - 7.52 (m, 3H), 7.32 (t, J = 7.8 Hz, 1H), 6.85 (s, 1H) , 6.33-6.36 (m, 2H), 5.65-5.69 (m, 1H), 3.93 (s, 3H), 2.93 (t, J = 6 Hz, 2H), 2.75 (s, 3H), 2.32 - 2.36 (m, 2H), 2.29 (s, 6H).

Example 8 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1H-indol-1-yl)thieno[3,2 -d]pyrimidine-2- Amino}phenyl}prop-2-enamide (Compound T-8)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 42.

Under a nitrogen atmosphere, a continuously stirred single-necked round bottom flask was sequentially charged with hydrazine (1.18 g, 10 mmol), compound 2 (2 g, 10 mmol), potassium carbonate (1.5 g, 11 mmol) and DMF (20 mL). After completion, the mixture was stirred at room temperature for 1 hour, and 20 mL of water was added to the reaction system. After filtration, the cake was washed with water and dried to give a white solid product (yield: 70%). LC-MS (APCI): m / z = 286 (M + 1) +.

Step 2 Synthesis of Compound 43.

Compound 42 (900 mg, 3.15 mmol), compound 4 (700 mg, 3.78 mmol), p-toluenesulfonic acid (651 mg, 3.78 mmol) and 2-pentanol (6 mL) were added to a single-neck round under nitrogen. After reacting for 3 hours at 105 ° C in a bottom flask, the cake was filtered and washed with 2-pentanol and water, and dried to give a brown solid product (1 g, yield: 73%). LC-MS (APCI): m / z = 436 (M + 1) +.

Step 3 Synthesis of Compound 44.

Under a nitrogen atmosphere, compound 43 (436 mg, 1 mmol), compound 6 (120 mg, 1.2 mmol), potassium carbonate (200 mg, 1.5 mmol) and DMF (10 mL) were added to a three-neck round bottom flask and stirred at room temperature. After an hour, it was diluted with 20 mL of water, filtered, washed with water, and the cake was dissolved in dichloromethane, and then extracted, and the organic phase was collected and purified by column chromatography to give a white solid product (yield: 77%). ¹ H NMR (300 MHz, CDCl ₃ ) (δ / ppm) 9.24 (s, 1H), 8.43 (d, J = 4.8 Hz, 1H), 7.95 (d, J = 2.4 Hz, 1H), 7.89 (d, J) =3.3 Hz, 1H), 7.66 (d, J = 4.8 Hz, 1H), 7.59 (s, 1H), 7.39 (d, J = 3 Hz, 1H), 7.33 - 7.35 (m, 1H), 7.26-7.29 ( m, 1H), 6.82 (d, J = 2.1 Hz, 1H), 6.69 (s, 1H), 3.97 (s, 3H), 3.28 (t, J = 4.2 Hz, 2H), 2.89 (s, 3H), 2.57 (t, J = 4.5 Hz, 2H), 2.27 (s, 6H). LC-MS (APCI): m / z = 518 (M + 1) +.

Step 4 Synthesis of Compound 45.

Compound 44 (371 mg, 0.7 mmol), iron powder (235 mg, 4.2 mmol) and ammonium chloride (37 mg, 0.7 mmol) were added to a mixture of water/ethanol (10 mL / 2 mL) in a nitrogen atmosphere, and then heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed and water was added, and extracted with dichloromethane. The organic phase was collected to give 245 mg of product as a yellow solid. LC-MS (APCI): m / z = 488 (M + 1) +.

Step 5 Synthesis of Compound T-8.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 50 mL of dichloromethane was added to the compound 45 (245 mg, 0.5 mmol), and after it was completely dissolved, triethylamine (75 mg, 0.75 mmol) was added dropwise. The system was cooled to -20 ° C, and the compound 9 (54 mg, 0.6 mmol) was slowly added, and the stirring was continued for 15 minutes. The reaction was quenched with water and the organic phase was separated and washed with water and saturated sodium hydrogen carbonate solution. Chromatography gave 166 mg of product as a yellow solid, yield 61%. LC-MS (APCI): m / z = 542 (M + 1) +. ¹ H NMR (300 MHz, DMSO-d ₆ ) (δ/ppm) 10.08 (s, 1H), 9.75 (s, 1H), 8.44 (d, J = 8.4 Hz, 1H), 8.02 (d, J = 3.6 Hz) , 1H), 7.89 (d, J = 5.4 Hz, 1H), 7.65-7.68 (m, 2H), 7.55 (d, J = 5.4 Hz, 1H), 7.25-7.35 (m, 2H), 6.82 (d, J=4.2 Hz, 2H), 6.40-6.42 (m, 2H), 5.67-5.72 (m, 1H), 3.91 (s, 3H), 2.92 (t, J = 5.1 Hz, 2H), 2.73 (s, 3H) ), 2.32 - 2.36 (m, 2H), 2.30 (s, 6H).

Example 9 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(3-methyl-1H-indazol-1-yl)thiophene And [3,2-d] Pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-9)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 47.

Compound 46 (0.66 g, 5 mmol), compound 2 (1 g, 5 mmol), potassium carbonate (0.83 g, 6 mmol) and DMF (10 mL) were added sequentially to a continuously stirred single-necked round bottom flask under nitrogen. After completion, the mixture was stirred at room temperature for 1 hour, and 20 mL of water was added to the reaction system. After filtration, the cake was washed with water and dried to give a white solid product (1.2 g, yield: 80%). LC-MS (APCI): m / z = 301 (M + 1) +.

Step 2 Synthesis of Compound 48.

Compound 47 (1.2 g, 4 mmol), Compound 4 (700 mg, 3.78 mmol), p-toluenesulfonic acid (827 mg, 4.8 mmol) and 2-pentanol (6 mL) were added to a single-neck round under nitrogen. After reacting for 3 hours at 105 ° C in a bottom flask, the cake was filtered and washed with 2-pentanol and water, and dried to give a brown solid product (1.1 g, yield 61%). LC-MS (APCI): m / z = 451 (M + 1) +.

Step 3 Synthesis of Compound 49.

Compound 48 (450 mg, 1 mmol), compound 6 (120 mg, 1.2 mmol), potassium carbonate (200 mg, 1.5 mmol) and DMF (10 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After an hour, it was diluted with 20 mL of water, filtered, washed with water, and the filter cake was dissolved in dichloromethane, and then extracted, and the organic phase was collected and purified by column chromatography to yield product (yield: LC-MS (APCI): m / z = 532 (M + 1) +.

Step 4 Synthesis of Compound 50.

Compound 49 (160 mg, 0.3 mmol), iron powder (100 mg, 1.8 mmol) and ammonium chloride (16 mg, 0.3 mmol) were added to a mixture of water/ethanol (10 mL / 2 mL) in a nitrogen atmosphere, and then heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and then the organic phase was collected to give 150 mg of product as a yellow solid. LC-MS (APCI): m / z = 503 (M + 1) +.

Step 5 Synthesis of Compound T-9.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 50 mL of dichloromethane was added with a compound 50 (150 mg, 0.3 mmol). After it was completely dissolved, triethylamine (50 mg, 0.45 mmol) was added dropwise. The system was cooled to -20 ° C, and the compound 9 (33 mg, 0.36 mmol) was slowly added and stirring was continued for 15 minutes. The reaction was quenched with water, and the organic phase was separated and washed with water and saturated sodium hydrogen carbonate solution. Chromatography gave a yellow solid product (yield: 86). LC-MS (APCI): m / z = 557 (M + 1) +. ^{1 H NMR (300MHz, DMSO-} d 6) (δ / ppm) 10.05 (s, 1H), 9.66 (s, 1H), 8.85 (d, J = 5.1Hz, 1H), 7.89 (d, J = 3.6Hz , 1H), 7.68 (d, J = 4.8 Hz, 1H), 7.42 - 7.48 (m, 3H), 7.30 (t, J = 4.2 Hz, 1H), 6.83 (s, 1H), 6.36 (d, J = 3.3 Hz, 2H), 5.65-5.68 (m, 1H), 3.91 (s, 3H), 2.93 (t, J = 3.3 Hz, 2H), 2.74 (s, 3H), 2.69 (s, 3H), 2.36 ( s, 2H), 2.31 (s, 6H).

Example 10 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(3-methyl-1H-indol-1-yl)thiophene And [3,2-d] Pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-10)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 52.

Compound 51 (1.28 g, 9.75 mmol), Compound 2 (2 g, 9.75 mmol), potassium carbonate (2.02 g, 14.63 mmol) and DMF (20 mL) were sequentially added to a continuously stirred single-necked round bottom flask under nitrogen. After the completion of the addition, the mixture was stirred at room temperature overnight, and 20 mL of water was added to the reaction system. After filtration, the cake was washed with water and dried to give a brown solid product (2.2 g, yield: 75%). LC-MS (APCI): m / z = 300 (M + 1) +. ¹ H NMR (300 MHz, CDCl ₃ ) (δ / ppm) 8.70 (d, J = 8.1 Hz, 1H), 8.05 (d, J = 5.4 Hz, 1H), 7.77 (d, J = 1.2 Hz, 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.55 (d, J = 5.4 Hz, 1H), 7.46-7.41 (m, 1H), 7.37-7.32 (m, 1H), 2.41 (d, J = 1.2 Hz, 3H).

Step 2 Synthesis of Compound 53.

Compound 52 (966 mg, 3.2 mmol), Compound 4 (500 mg, 2.7 mmol), p-toluenesulfonic acid (555 mg, 3.2 mmol) and 2-pentanol (6 mL) were added to a single-neck round under nitrogen. After reacting at 105 ° C for 3 hours in a bottom flask, the cake was filtered and washed sequentially with 2-pentanol and water, and dried to give a brown solid product 950 mg (yield: 79%). LC-MS (APCI): m / z = 450 (M + 1) +.

Step 3 Synthesis of Compound 54.

Compound 53 (500 mg, 1.1 mmol), compound 6 (170 mg, 1.7 mmol), potassium carbonate (461 mg, 3.3 mmol) and DMF (10 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After 3 hours, it was diluted with 20 mL of water, filtered, washed with water and the cake was dissolved in dichloromethane, and then extracted, and the organic phase was collected and purified by column chromatography to give the product as a yellow solid, 220 mg, yield 37%. LC-MS (APCI): m / z = 532 (M + 1) +. ¹ H NMR (300 MHz, CDCl ₃ ) (δ / ppm) 9.26 (s, 1H), 8.46 (d, J = 5.1 Hz, 1H), 7.92 (d, J = 3.3 Hz, 1H), 7.77 (d, J = 0.6 Hz, 1H), 7.61 (d, J = 4.8 Hz, 1H), 7.58 (s, 1H), 7.41 (d, J = 3.3 Hz, 1H), 7.37-7.35 (m, 1H), 7.32-7.30 ( m, 1H), 6.71 (s, 1H), 4.01 (s, 3H), 3.29 (t, J = 3.9 Hz, 2H), 2.90 (s, 3H), 2.58 (t, J = 4.2 Hz, 2H), 2.42 (d, J = 0.9 Hz, 3H), 2.28 (s, 6H).

Step 4 Synthesis of Compound 55.

Under a nitrogen atmosphere, compound 54 (210 mg, 0.4 mmol), iron powder (110 mg, 2.0 mmol) and ammonium chloride (42 mg, 0.8 mmol) were added to a mixture of water/ethanol (10 mL / 2 mL) and heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, and the organic phase was collected by dichloromethane. LC-MS (APCI): m / z = 502 (M + 1) +.

Step 5 Synthesis of Compound T-10.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 30 mL of dichloromethane was successively charged with compound 55 (198 mg, 0.4 mmol). After it was completely dissolved, triethylamine (80 mg, 0.8 mmol) was added dropwise to react. The system was cooled to -10 ° C, and a solution of compound 9 (39 mg, 0.4 mmol) in dichloromethane (1 mL) was slowly added, and stirring was continued for 30 minutes. The reaction was quenched with water and the organic phase was separated, and water and saturated sodium hydrogen carbonate solution After washing, the organic phase was collected and purified by column chromatography toield of pale yellow solid product (yield: 39%). LC-MS (APCI): m / z = 556 (M + 1) +. ^{1 H NMR (300MHz, DMSO-} d 6) (δ / ppm) 10.13 (s, 1H), 9.82 (s, 1H), 8.48 (s, 1H), 8.45 (d, J = 4.5Hz, 1H), 8.32 (d, J = 5.1 Hz, 1H), 7.90 (s, 1H), 7.59-7.58 (m, 1H), 7.33 (d, J = 3 Hz, 1H), 7.23-7.19 (m, 2H), 6.99 (s , 1H), 6.23 (dd, J = 1.2 Hz, 10.5 Hz, 1H), 5.71 (dd, J ==1.2 Hz, 7.2 Hz, 1H), 3.82 (s, 3H), 3.30 (s, 2H), 2.68 -2.61 (m, 5H), 2.49 (s, 6H), 2.35 (s, 3H).

Example 11 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(imidazo[1,2-a]pyridin-3-yl) Thiophene [3,2-d]pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-11)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 58.

A mixed solution of compound 56 (400 mg, 1.49 mmol) and N-bromosuccinimide (NBS, 318 mg, 1.79 mmol) in 1,4-dioxane/water (5 mL / 1 mL) was stirred at room temperature After 1 hour, the compound 57 (140 mg, 1.49 mmol) was added, and the mixture was heated to 85 ° C and reacted at this temperature for 2 hours, cooled to room temperature, filtered, washed with water and dried to give a yellow solid product (yield: 73%). LC-MS (APCI): m / z = 287.3 (M + 1) +. ¹ H NMR (300 MHz, CDCl ₃ ) (δ / ppm) 8.89 (d, J = 9 Hz, 1H), 8.79 (s, 1H), 8.63 (d, J = 6.9 Hz, 1H), 8.04 (d, J = 4.5) Hz, 1H), 7.58-7.55 (m, 2H), 7.12-7.06 (m, 1H).

Step 2 Synthesis of Compound 59.

Compound 58 (308 mg, 1.1 mmol), Compound 4 (200 mg, 1.1 mmol), p-toluenesulfonic acid (555 mg, 3.2 mmol) and 2-pentanol (10 mL) were added to a single-neck round under nitrogen. After reacting at 105 ° C for 16 hours in a bottom flask, after cooling to room temperature, 10 mL of a saturated sodium hydrogencarbonate solution was added, and the mixture was stirred for 30 minutes, then filtered and washed with water, and dried to give a brown solid product (yield: 64%). LC-MS (APCI): m / z = 437 (M + 1) +.

Step 3 Synthesis of Compound 60.

Compound 59 (300 mg, 0.7 mmol), compound 6 (84 mg, 0.8 mmol), potassium carbonate (285 mg, 2.1 mmol) and DMF (10 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and filtered, and then filtered, and the organic phase was collected and purified by column chromatography to give 148 mg of product as an orange solid. LC-MS (APCI): m / z = 519 (M + 1) +.

Step 4 Synthesis of Compound 61.

Compound 60 (148 mg, 0.3 mmol), iron powder (159 mg, 2.8 mmol) and ammonium chloride (76 mg, 1.4 mmol) were sequentially added to a mixture of water/ethanol (6 mL / 3 mL) under a nitrogen atmosphere, and heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, extracted with dichloromethane, and the organic phase was collected to give 139 mg of solid product, yield 99%. LC-MS (APCI): m / z = 488 (M + 1) +. ¹ H NMR (300 MHz, CDCl ₃ ) (δ / ppm) 9.91 (d, J = 6.6 Hz, 1H), 9.10 (s, 1H), 8.64 (s, 1H), 7.93 (d, J = 5.4 Hz, 1H) , 7.79 (d, J = 9 Hz, 1H), 7.51 (s, 1H), 7.48-7.41 (m, 1H), 7.40 (d, J = 5.4 Hz, 1H), 7.08-7.03 (m, 1H), 6.73 (s, 1H), 4.02 (s, 3H), 3.32 (t, J = 6.9 Hz, 2H), 2.91 (s, 3H), 2.62 (t, J = 7.2 Hz, 2H), 2.31 (s, 6H) .

Step 5 Synthesis of Compound T-11.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 30 mL of dichloromethane was added sequentially to compound 61 (139 mg, 0.3 mmol). After it was completely dissolved, triethylamine (57 mg, 0.6 mmol) was added dropwise. The system was cooled to -10 ° C. Compound 9 (31 mg, 0.3 mmol) was slowly added and stirring was continued for 15 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 75 mg, the yield was 49%. LC-MS (APCI): m / z = 543 (M + 1) +. ¹ H NMR (300 MHz, DMSO-d ₆ ) (δ / ppm) 9.98 (d, J = 6.9 Hz, 1H), 8.49 - 8.47 (m, 2H), 8.10 (d, J = 5.7 Hz, 1H), 7.70 (d, J = 6.3 Hz, 1H), 7.54 - 7.49 (m, 1H), 7.29 (d, J = 5.7 Hz, 1H), 7.08 (t, J = 6.6 Hz, 1H), 6.97 (s, 1H) , 6.62-6.53 (m, 1H), 6.47-6.41 (m, 1H), 5.87-5.83 (m, 1H), 3.98 (s, 3H), 3.44 (t, J = 5.7 Hz, 2H), 3.19 (t , J = 5.4 Hz, 2H), 2.81 (s, 6H), 2.71 (s, 3H).

Example 12 N-(2-((2-(Dimethylamino)ethyl)(methyl)amino)-5-((7-(hydroxymethyl)-4-(1-methyl-1H-)吲哚-3-yl)-7H-pyridyl [2,3-d]pyrimidin-2-yl)amino)-4-methoxyphenyl)prop-2-enamide (Compound T-12)

Example 13 N-(2-((2-(Dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-((4-(1-methyl-1H-indole) -3-yl)-7H-pyridyl [2,3-d]pyrimidin-2-yl)amino)phenyl)prop-2-enamide (Compound T-13)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 63.

Anhydrous aluminum trichloride (4.0 g, 30.0 mmol) was added portionwise to a mixture of compound 62 (1.9 g, 10.0 mmol) and 1-methylindole (2.6 g, 20.0 mmol) under nitrogen. Water in 1,2-dichloroethane (DCE, 100 mL). The reaction was stirred at reflux overnight and the reaction was cooled to room temperature. The reaction was quenched with EtOAc EtOAc (EtOAc m. It is 32%. LC-MS (APCI): m/z = </ ^RTI>

Step 2 Synthesis of Compound 64.

Sodium hydride (38 mg, 1.60 mmol) was added to a solution of compound 63 (300 mg, 1.1 mmol) in anhydrous N,N-dimethylformamide (5mL), and the reaction was warmed to room temperature and stirred for 30 minutes. Further, 2-(trimethylsilyl)ethoxymethyl chloride (SEMCl, 211 mg, 1.3 mmol) was added dropwise to the reaction mixture under ice-cooling, and the mixture was stirred at room temperature for 2 hours. The reaction was extracted with water (25 mL), EtOAc (EtOAc)EtOAc. The solid was 270 mg in a yield of 62%. LC-MS (APCI): m / z = 413 (M + 1) +.

Step 3 Synthesis of Compound 66.

Anhydrous tert-butanol (3 mL) was added to compound 64 (50 mg, 0.12 mmol) and compound 65 (45 mg, 0.15 mmol), potassium carbonate (50 mg, 0.36 mmol), Pd ₂ (dba) ₃ (10 mg, In a mixture of 0.01 mmol) and X-Phos (10 mg, 0.02 mmol), the reaction was stirred under a nitrogen atmosphere and stirred at 90 ° C overnight. The mixture was cooled to room temperature, filtered over Celite, and filtered, and filtered, and the filtrate was concentrated under reduced pressure, and concentrated to give a pale yellow solid, 45 mg, yield: 56.1%. LC-MS (APCI): m/z = </ ^RTI>

Step 4 Synthesis of the compounds T-12 and T-13.

Trifluoroacetic acid (3 mL) was added to a solution of compound 66 (160 mg, 0.24 mmol) in dichloromethane (6 mL), and the reaction was allowed to react at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure. EtOAc EtOAc m. The reaction mixture was concentrated under reduced pressure, diluted with water (20 mL), dichloromethane (30 mL?? The mixture was further purified by thin layer chromatography (DCM/MeOH v/v = 15/1) to give N-(2-((2-dimethylamino)ethyl) (methyl)amino)-5- (( 7-(Hydroxymethyl)-4-(1-methyl-1H-indol-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)-4-methoxy Phenyl phenyl) acrylamide (Compound T-12) 50 mg, yield 37%. LC-MS (APCI): m / z = 569 (M + 1) +. ¹ H NMR (300 MHz, DMSO-d ₆ ) (δ/ppm) 10.07 (s, 1H), 9.44 (s, 1H), 8.59 (d, J = 7.7 Hz, 1H), 8.44 (s, 1H), 7.72 (s, 1H), 7.51 (d, J = 8.0 Hz, 1H), 7.33 (d, J = 3.7 Hz, 1H), 7.28 - 7.13 (m, 2H), 7.01 (s, 1H), 6.92 (d, J = 3.7 Hz, 1H), 6.58 (br, 1H), 6.44 (t, J = 7.3 Hz, 1H), 6.24 (d, J = 15.3 Hz, 1H), 5.77 - 5.71 (m, 1H), 5.64 ( d, J = 7.2 Hz, 2H), 3.93 (s, 3H), 3.91 (s, 3H), 3.11 - 2.93 (m, 2H), 2.68 (s, 3H), 2.51 - 2.49 (m, 2H), 2.38 (s, 6H).

A yellow solid mixture was obtained which was further purified by thin layer chromatography (DCM/MeOH v/v=15/1) to afford N-(2-((2-dimethylamino)ethyl)(methyl)amino)- 4-methoxy-5-((6-(1-methyl-1H-indol-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)phenyl) Acrylamide (Compound T-13) 30 mg, yield 23.%. LC-MS (APCI): m/z = </ ^{RTI> 1 H NMR (300MHz, DMSO-} d 6) (δ / ppm) 11.39 (s, 1H), 10.03 (s, 1H), 8.79 (s, 1H), 8.58 (d, J = 7.9Hz, 1H), 8.41 (s, 1H), 7.65 (s, 1H), 7.49 (d, J = 8.1 Hz, 1H), 7.25 - 7.18 (m, 1H), 7.17 - 7.09 (m, 2H), 6.98 (s, 1H), 6.83 (d, J = 1.9 Hz, 1H), 6.54 (s, 1H), 6.20 (d, J = 16.7 Hz, 1H), 5.72 (d, J = 10.6 Hz, 1H), 3.92 (s, 3H), 3.86 (s, 3H), 3.09 - 2.96 (m, 2H), 2.67 (s, 3H), 2.50 - 2.48 (m, 2H), 2.38 (s, 6H).

Example 14 N-(2-((2-(Dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-((4-(1-methyl-1H-indole) -3-yl)-1H-pyridyl Zoxa[3,4-d]pyrimidin-6-yl)amino)phenyl)prop-2-enamide (Compound T-14)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 68.

Anhydrous aluminum trichloride (4.00 g, 30.0 mmol) was added portionwise to compound 67 (1.89 g, 10.0 mmol) and 1-methyl-1H-indole (1.38 g, 1.05 mmol). ) in anhydrous DCE (70 mL). The reaction was stirred at reflux overnight and the reaction was cooled to room temperature. The reaction was quenched with EtOAc EtOAc (EtOAc m. It is 32%. LC-MS (APCI): m / z = 284 (M + 1) +.

Step 2 Synthesis of Compound 69.

Sodium hydride (101 mg, 4.20 mmol) was added to a solution of compound 68 (900 mg, 2.8 mmol) in anhydrous N, N-dimethylformamide (15 mL), and the mixture was warmed to room temperature and stirred for 1 hour. Further, 2-(trimethylsilyl)ethoxymethyl chloride (570 mg, 3.40 mmol) was added dropwise to the reaction mixture under ice-cooling, and the mixture was stirred at room temperature overnight. The reaction was extracted with water (25 mL), EtOAc (EtOAc)EtOAc. The solid was 300 mg in a yield of 26%. LC-MS (APCI): m / z = 414 (M + 1) +.

Step 3 Synthesis of Compound 70.

Anhydrous tert-butanol (6 mL) was added to compound 69 (270 mg, 0.65 mmol) and compound 65 (210 mg, 0.78 mmol), potassium carbonate (278 mg, 2.00 mmol), Pd ₂ (dba) ₃ (60 mg, In a mixture of 0.06 mmol) and X-Phos (60 mg, 0.12 mmol), the reaction was stirred under nitrogen and the mixture was stirred at 90 ° C overnight. The mixture was cooled to room temperature, filtered over Celite, and filtered, and the filtrate was washed with methylene chloride. The filtrate was concentrated under reduced pressure, and the mixture was concentrated on a column (eluent: methylene chloride/methanol as a pale yellow solid to afford 180 mg, yield 41%. - MS (APCI): m/z = 670.3 (M + 1) ⁺ .

Step 4 Synthesis of Compound T-14.

Trifluoroacetic acid (3 mL) was added to a solution of compound 70 (180 mg, 0.27 mmol) in dichloromethane (6 mL). The reaction mixture was concentrated under reduced pressure. EtOAc EtOAc m. The reaction mixture was concentrated under reduced pressure, diluted with water (20 mL), dichloromethane (30 mL, EtOAc) The solid 50mg, yield 34%, LC-MS (APCI ): m / z = 540 (m + 1) +. ¹ H NMR (300 MHz, DMSO-d ₆ ) (δ/ppm) 13.12 (s, 1H), 9.97 (s, 1H), 8.72 - 8.58 (m, 2H), 8.55 - 8.37 (m, 2H), 8.28 ( s, 1H), 7.53 (d, J = 8.2 Hz, 1H), 7.30 - 7.12 (m, 2H), 6.98 (s, 1H), 6.78 (br, 1H), 6.21 (d, J = 16.8 Hz, 1H) ), 5.71 (d, J = 11.8 Hz, 1H), 3.94 (s, 3H), 3.83 (s, 3H), 3.16 (d, J = 4.6 Hz, 2H), 2.66 (s, 3H), 2.57 - 2.53 (m, 2H), 2.49 (s, 6H).

Example 15 N-(2-((2-(Dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-((6-(1-methyl-1H-indole) -3-yl)-9H-嘌 Indole-2-yl)amino)phenyl)prop-2-enamide (Compound T-15)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 72.

Anhydrous aluminum trichloride (4.00 g, 30.0 mmol) was added portionwise to a mixture of compound 71 (1.89 g, 10.0 mmol) and 1-methylindole (1.57 g, 12.0 mmol) under nitrogen. Water in DCE (100 mL). The reaction was stirred at reflux overnight and the reaction was cooled to room temperature. The reaction was quenched with EtOAc EtOAc (EtOAc m. The rate is 71%. LC-MS (APCI): m / z = 284.1 (M + 1) +.

Step 2 Synthesis of Compound 73.

3,4-Dihydro-2H-pyran (THP, 444 mg, 5.28 mmol) was added to compound 72 (300 mg, 1.05 mmol) and anhydrous p-toluenesulfonic acid (30 mg, 0.16 mmol) of tetrahydrofuran. In a 10 mL) solution, the reaction was stirred under reflux overnight. The organic layer was concentrated under reduced pressure and the solvent was evaporated. LC-MS (APCI): m / z = 368.1 (M + 1) +.

Step 3 Synthesis of Compound 74.

Anhydrous tert-butanol (10 mL) was added to compound 73 (210 mg, 0.57 mmol) and compound 65 (200 mg, 0.68 mmol), potassium carbonate (190 mg, 1.4 mmol), Pd ₂ (dba) ₃ (55 mg, In a mixture of 0.06 mmol) and X-Phos (56 mg, 0.12 mmol), the reaction was stirred under nitrogen and the mixture was stirred at 90 ° C overnight. The mixture was cooled to room temperature, filtered over Celite, and filtered, and filtered, and the filtrate was concentrated under reduced pressure, and concentrated to give a pale yellow solid (yield: 56%). LC-MS (APCI): m / z = 624.3 (M + 1) +.

Step 4 Synthesis of Compound T-15.

Under N ₂ protection, 4M Hydrogen chloride in methanol (2mL) was added to the compound in tetrahydrofuran (5mL) 74 (200mg, 0.3mmol ) was, reaction at room temperature for 1 hour. The pH was adjusted to basic with saturated sodium bicarbonate, and extracted with dichloromethane. The organic phase was combined and dried over anhydrous sodium sulfate. The solvent was removed, and the concentrate was subjected to column separation to obtain a yellow solid 120 mg, yield 75%, LC-MS (APCI) ): m/z = 540.3 (M + 1) ⁺ . ¹ H NMR (300 MHz, DMSO-d ₆ ) (δ/ppm) 12.76 (s, 1H), 10.11 (s, 1H), 8.95 (s, 1H), 8.77 (s, 1H), 8.69 (d, J = 7.8 Hz, 1H), 8.10 (s, 1H), 7.93 (s, 1H), 7.51 (d, J = 8.1 Hz, 1H), 7.23 (t, J = 7.2 Hz, 1H), 7.12 (t, J = 7.3 Hz, 1H), 7.02 (s, 1H), 6.42 (dd, J = 17.0, 10.0 Hz, 1H), 6.18 (dd, J = 16.9, 2.0 Hz, 1H), 5.72 (dd, J = 10.1, 2.0 Hz, 1H), 3.94 (s, 3H), 3.83 (s, 3H), 2.98 - 2.86 (m, 2H), 2.72 (s, 3H), 2.44 - 2.30 (m, 2H), 2.24 (s, 6H) .

Example 16 N-(2-((2-(Dimethylamino)ethyl)(methyl)amino)-4-methoxy-5-((9-methyl-6-(1-methyl) -1H-吲哚-3- Base)-9H-indol-2-yl)amino)phenyl)prop-2-enamide (Compound T-16)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 75.

Iodomethane (960 mg, 6.76 mmol) was added to a solution of compound 72 (300 mg, 1.06 mmol) and potassium carbonate (30 mg, 0.16 mmol) in anhydrous N,N-dimethylformamide (5 mL). The reaction was carried out in an ice bath for 3 hours. The reaction was extracted with water (25 mL), EtOAc (EtOAc (EtOAc) The yellow solid was 190 mg in a yield of 61%. LC-MS (APCI): m / z = 98.1 (M + 1) +.

Step 2 Synthesis of Compound T-16.

Anhydrous tert-butanol (6 mL) was added to compound 75 (190 mg, 0.64 mmol) and compound 65 (210 mg, 0.7 mmol), potassium carbonate (278 mg, 2.0 mmol), Pd ₂ (dba) ₃ (60 mg, In a mixture of 0.06 mmol) and X-Phos (0.12 mg, 0.12 mmol), the reaction was stirred under nitrogen atmosphere and stirred at 90 ° C overnight. After cooling to room temperature, the mixture was filtered through Celite, and filtered, and filtered, and the filtrate was concentrated under reduced pressure, and concentrated to give a pale yellow solid (yield: 35%). LC-MS (APCI): m / z = 554.3 (M + 1) +. ¹ H NMR (300 MHz, DMSO-d ₆ ) (δ/ppm) 9.95 (s, 1H), 9.28 (s, 1H), 8.95 (s, 1H), 8.68 (d, J = 7.8 Hz, 1H), 8.16 (s, 1H), 7.94 (s, 1H), 7.55 - 7.49 (m, 1H), 7.28 - 7.13 (m, 2H), 7.01 (s, 1H), 6.84 - 6.63 (m, 1H), 6.28 - 6.10 (m, 1H), 5.76–5.67 (m, 1H), 3.94 (s, 3H), 3.88 (s, 3H), 3.82 (s, 3H), 3.15–2.99 (m, 2H), 2.67 (s, 3H) ), 2.54–2.46 (m, 8H).

Example 17 N-(5-((6-(5,6-Dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-9H-indol-2-yl)amino )-2-((2-(dimethylamide) Ethyl)ethyl (meth)amino)-4-methoxyphenyl)prop-2-enamide (Compound T-17)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 76.

Anhydrous aluminum trichloride (4.00 g, 30.0 mmol) was added portionwise to compound 71 (1.89 g, 10.0 mmol) and 5,6-dihydro-4H-pyrrolo[3,2 at room temperature under nitrogen. , 1-ij] quinoline (1.65 g, 10.5 mmol) in anhydrous DCE (100 mL). The reaction was stirred at reflux overnight and the reaction was cooled to room temperature. The reaction was quenched with EtOAc EtOAc (EtOAc m. The rate is 55%. LC-MS (APCI): m / z = 310.1 (M + 1) +.

Step 2 Synthesis of Compound 77

3,4-Dihydro-2H-pyran (165 mg, 1.95 mmol) was added to a solution of compound 76 (500 mg, 1.62 mmol) in tetrahydrofuran (10 mL), and the reaction was stirred under reflux overnight. The organic layer was concentrated under reduced pressure, and the solvent was evaporated. LC-MS (APCI): m / z = 394.2 (M + 1) +.

Step 3 Synthesis of Compound 78.

Anhydrous tert-butanol (10 mL) was added to compound 77 (215 mg, 0.55 mmol) and compound 65 (180 mg, 0.62 mmol), potassium carbonate (230 mg, 1.65 mmol), Pd ₂ (dba) ₃ (50 mg, In a mixture of 0.05 mmol) and X-Phos (52 mg, 0.11 mmol), the reaction was stirred under a nitrogen atmosphere and stirred at 90 ° C overnight. The mixture was cooled to room temperature, filtered over Celite, and filtered, and filtered, and the filtrate was concentrated under reduced pressure, and concentrated to give a pale yellow solid (yield: 50%). LC-MS (APCI): m / z = 650.4 (M + 1) +.

Step 4 Synthesis of Compound T-17.

Under N ₂ protection, 4M Hydrogen chloride in methanol (2mL) was added in tetrahydrofuran (5mL) compound 78 (200mg, 0.3mmol) was, reaction at room temperature for 1 hour. The pH was adjusted to basic with saturated sodium bicarbonate, and extracted with dichloromethane (30 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. The solvent was removed and concentrated to give a beige solid (yield: . LC-MS (APCI): m / z = 566.4 (M + 1) +. ^{1 H NMR (300MHz, DMSO-} d 6) (δ / ppm) 12.74 (s, 1H), 10.12 (s, 1H), 8.94 (s, 1H), 8.80 (s, 1H), 8.36 (d, J = 7.8 Hz, 1H), 8.09 (s, 1H), 7.88 (s, 1H), 7.05–6.98 (m, 2H), 6.96–6.90 (m, 1H), 6.42 (dd, J = 16.9, 10.1 Hz, 1H) ), 6.19 (dd, J = 17.0, 1.6 Hz, 1H), 5.72 (d, J = 11.7 Hz, 1H), 4.44 - 4.22 (m, 2H), 3.83 (s, 3H), 3.02 - 2.84 (m, 4H), 2.72 (s, 3H), 2.42 - 2.30 (m, 2H), 2.24 (s, 6H), 2.20 - 2.11 (m, 2H).

Example 18 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(imidazo[1,2-a]pyrimidin-3-yl) Thiophene [3,2-d]pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-18)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 81.

A mixed solution of compound 56 (400 mg, 1.49 mmol) and N-bromosuccinimide (NBS, 318 mg, 1.79 mmol) in 1,4-dioxane/water (5 mL / 1 mL) was stirred at room temperature After 1 hour, the compound 80 (140 mg, 1.49 mmol) was added, and the mixture was heated to 85 ° C and reacted at this temperature for 2 hours, cooled to room temperature and then filtered, washed with water and dried to give a yellow solid product (yield: 70.1%). LC-MS (APCI): m / z = 288.3 (M + 1) +.

Step 2 Synthesis of Compound 82.

Compound 81 (161 mg, 559 umol), Compound 4 (104 mg, 559 umol), p-toluenesulfonic acid (425 mg, 2.23 mmol) and 2-pentanol (5 mL) were added to a single-necked round bottom flask under nitrogen. After reacting at 105 ° C for 16 hours, after cooling to room temperature, 10 mL of a saturated sodium hydrogencarbonate solution was added, and after stirring for 30 minutes, the cake was filtered and washed with water, and dried to give a brown solid product (yield: 40.9%). LC-MS (APCI): m / z = 438 (M + 1) +.

Step 3 Synthesis of Compound 83.

Under a nitrogen atmosphere, compound 82 (100 mg, 0.229 mmol), compound 6 (70 mg, 0.686 mmol), potassium carbonate (95 mg, 0.686 mmol) and DMF (3 mL) were added to a three-neck round bottom flask and stirred at room temperature. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and the cake was dissolved in dichloromethane, and then extracted. The organic phase was collected and purified by column chromatography to give 40 mg of the product as an orange solid. LC-MS (APCI): m / z = 520.4 (M + 1) +.

Step 4 Synthesis of Compound 84.

Compound 83 (40 mg, 0.077 mmol), iron powder (43 mg, 0.77 mmol) and ammonium chloride (21 mg, 0.0385 mmol) were added to a mixture of water/ethanol (3 mL / 1 mL) in a nitrogen atmosphere, and then heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and the organic phase was collected to give a solid product (yield: 82.2%). LC-MS (APCI): m / z = 490.2 (M + 1) +.

Step 5 Synthesis of Compound T-18.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 5 mL of dichloromethane was added sequentially to compound 84 (31 mg, 0.063 mmol). After it was completely dissolved, triethylamine (9.6 mg, 0.095 mmol) was added dropwise. The reaction system was cooled to -10 ° C, and the compound 9 (7 mg, 0.076 mmol) was slowly added and stirring was continued for 15 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid. The product was 11 mg in a yield of 32.0%. LC-MS (APCI): m / z = 544.2 (M + 1) +. ^{1 H NMR (300MHz, MeOH-} D4) (δ / ppm) 9.14 (s, 1H), 9.08 (s, 1H), 8.69 (dd, J = 9.9Hz, J = 0.9Hz, 1H), 8.53 (s, 1H), 7.14 (d, J = 3.3 Hz, 1H), 7.20 (d, J = 3.3 Hz, 1H), 7.04 (d, J = 4.2 Hz, 1H), 6.90 (s, 1H), 6.57-6.51 ( m,1H), 6.37 (dd, J=13.6 Hz, J=1.2 Hz, 1H), 6.16 (s, 1H), 5.79-5.77 (m, 1H), 3.95 (s, 3H), 3.23 (t, J =3.3 Hz, 2H), 2.90 (t, J = 3.3 Hz, 2H), 2.72 (s, 3H), 2.62 (s, 6H).

Example 19 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(imidazo[1,2-a]pyrazin-3-yl) Thiophene [3,2-d]pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-19)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 86.

A mixed solution of compound 56 (400 mg, 1.49 mmol) and N-bromosuccinimide (NBS, 318 mg, 1.79 mmol) in 1,4-dioxane/water (5 mL / 1 mL) was stirred at room temperature After 1 hour, the compound 85 (140 mg, 1.49 mmol) was added, and the mixture was heated to 85 ° C and reacted at this temperature for 2 hours, cooled to room temperature, filtered, and washed with water and dried to yield 255 mg of product as a yellow solid. LC-MS (APCI): m / z = 288.3 (M + 1) +.

Step 2 Synthesis of Compound 87.

Compound 86 (255 mg, 886 umol), Compound 4 (150 mg, 806 umol), p-toluenesulfonic acid (28 mg, 3.3 mmol) and 2-pentanol (5 mL) were sequentially added to a single-necked round bottom flask under nitrogen. After reacting at 105 ° C for 16 hours, after cooling to room temperature, 10 mL of a saturated sodium hydrogencarbonate solution was added, and after stirring for 30 minutes, the cake was filtered and washed with water, and dried to give a brown solid product (yield: 28.4%). LC-MS (APCI): m / z = 438 (M + 1) +.

Step 3 Synthesis of Compound 88.

Under a nitrogen atmosphere, compound 82 (100 mg, 0.229 mmol), compound 6 (70 mg, 0.686 mmol), potassium carbonate (95 mg, 0.686 mmol) and DMF (3 mL) were added to a three-neck round bottom flask and stirred at room temperature. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and the cake was dissolved in dichloromethane, and then extracted. The organic phase was collected and purified by column chromatography to give 40 mg of the product as an orange solid. LC-MS (APCI): m / z = 520.4 (M + 1) +.

Step 4 Synthesis of Compound 89.

Compound 83 (40 mg, 0.077 mmol), iron powder (43 mg, 0.77 mmol) and ammonium chloride (21 mg, 0.0385 mmol) were added to a mixture of water/ethanol (3 mL / 1 mL) in a nitrogen atmosphere, and then heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and the organic phase was collected to give a solid product (yield: 82.2%). LC-MS (APCI): m / z = 490.2 (M + 1) +.

Step 5 Synthesis of Compound T-19.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 5 mL of dichloromethane was added sequentially to compound 84 (31 mg, 0.063 mmol). After it was completely dissolved, triethylamine (9.6 mg, 0.095 mmol) was added dropwise. The reaction system was cooled to -10 ° C, and the compound 9 (7 mg, 0.076 mmol) was slowly added and stirring was continued for 15 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid. The product was 11 mg in a yield of 32.0%. LC-MS (APCI): m / z = 544.2 (M + 1) +. ^{1 H NMR (500MHz, MeOH-} D4) (δ / ppm) 9.76 (d, J = 4.0Hz, 1H), 9.07 (s.1H), 8.56 (s, 1H), 8.50 (s, 1H), 8.05 ( d, J = 5.5 Hz, 1H), 7.94 (d, J = 5.0 Hz, 1H), 7.18 (d, J = 5.5 Hz, 1H), 6.96 - 6.91 (m, 1H), 6.60 - 6.55 (m, 1H) ), 6.42-6.38 (m, 1H), 5.85-5.82 (m, 1H), 3.90 (s, 3H), 3.32-3.28 (m, 2H), 3.27-3.24 (m, 2H), 2.72 (s, 3H) ), 2.62 (s, 6H).

Example 20 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(2-methyl-1-H-benzene[d]-imidazole -1-) Thiophene [3,2-d]pyrimidin-2-yl]amino}phenyl)prop-2-enamide (Compound T-20)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 91.

Potassium carbonate (2.7 g, 19.51 mmol), and compound 2 (2.0 g, 9.75 mmol) were sequentially added to a solution of Compound 90 (1.29 g, 9.75 mmol) in DMF (20M1). It was poured into water (200 mL), and a large amount of solid was precipitated, filtered, and dried in vacuo to give a white solid (1.1 g, yield: 37.5%). LC-MS (APCI): m / z = 301.1 (M + 1) +.

Step 2 Synthesis of Compound 92.

Compound 91 (614 mg, 2.04 mmol), Compound 4 (380 mg, 2.04 mmol), Cs ₂ CO ₃ (798 mg, 2.45 mmol) were added to 1,4-dioxane (10 mL) in vacuo. ₂ (dba) ₃ (93 mg, 102 umol) and xantphos (228 mg, 408 umol), and the reaction mixture was reacted at 100 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, ethyl acetate (30 mL) was added, filtered, and the filter cake was washed with water (30mL) and dried to give a yellow solid (500 mg, yield: 54.4%). LC-MS (APCI): m / z = 451.2M + 1) +, 1 H NMR (400MHz, DMSO-D6) (δ / ppm) 9.00 (s, 1H), 8.96 (d, J = 8.0Hz, 1H ), 8.47 (d, J = 5.6 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.52 (d, J = 5.2 Hz, 1H), 7.34 - 7.22 (M, 4h), 3.99 (s) , 3H), 2.62 (s, 3H).

Step 3 Synthesis of Compound 93.

Compound 92 (500 mg, 1.11 mmol), compound 6 (170 mg, 1.67 mmol), potassium carbonate (307 mg, 2.22 mmol) and DMF (5 mL) were added to a three-neck round bottom flask and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and filtered, and then filtered, and the organic phase was collected and purified by column chromatography to give an orange solid product (yield: 59.2%). LC-MS (APCI): m / z = 533.4 (M + 1) +, 1 H NMR (400MHz, CDCl3) (δ / ppm) 9.26 (s, 1H), 7.98 (d, J = 5.6Hz, 1H) , 7.80 (d, J = 8.0 Hz, 1H), 7.73 (s, 1H), 7.50 (d, J = 5.6 Hz, 1H), 7.36-7.32 (m, 1H), 7.26-7.24 (m, 2H), 6.70 (s, 1H), 3.99 (s, 3H), 3.29 (t, J = 6.8 Hz, 2H), 2.90 (s, 3H), 2.74 (s, 3H), 2.57 (t, J = 5.6 Hz, 2H) ), 2.27 (s, 6H).

Step 4 Synthesis of Compound 94.

Compound 83 (350 mg, 657 umol), iron powder (367 mg, 6.57 mmol) and ammonium chloride (176 mg, 3.29 mmol) were sequentially added to a mixture of ethanol/water (10 mL/5 mL) under a nitrogen atmosphere, and heated to reflux. After reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and then the organic phase was collected to obtain a solid product (yield: 99. LC-MS (APCI): m / z = 503.3 (M + 1) +.

Step 5 Synthesis of Compound T-20.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 5 mL of dichloromethane was added with compound 84 (330 mg, 656 umol). After it was completely dissolved, triethylamine (100 mg, 985 umol) was added dropwise to cool the reaction system. To a temperature of -10 ° C, compound 9 (71 mg, 788 um) was added slowly and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The rate is 43.8%. LC-MS (APCI): m / z = 557.2 (M + 1) +. ¹ H NMR (500 MHz, CDCl 3 ) (δ / ppm) 10.08 (br s, 1H), 9.72 (s, 1H), 7.92 (d, J = 5.5 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H) ), 7.75 (s, 1H), 7.57 (d, J = 6.0 Hz, 1H), 7.33-7.30 (m, 1H), 7.26-7.21 (m, 2H), 6.81 (s, 1H), 6.48-6.42 ( m, 1H), 6.38-6.32 (m, 1H), 5.69 (dd, J = 10.0 Hz, J = 1.5 Hz, 1H), 3.89 (s, 3H), 2.90 (t, J = 5.5 Hz, 2H), 2.73 (s, 3H), 2.71 (s, 3H), 2.32 (t, J = 5.5 Hz, 2H), 2.28 (s, 6H).

Example 21 N-(2-{2-Dimethylaminoethyl-methylamino}-5-(4-(4-fluoro-1-isopropyl-2-methyl-1Hbenzo[d]imidazole -6-) thiophene Benzo[3,2-d]pyrimidin-2-amino)-4-methoxyphenyl)acrylamide (T-21)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 96.

Under a nitrogen atmosphere, compound 95 (5.5 g, 20.37 mmol), decyl alcohol borate (10.3 g, 40.74 mmol), potassium acetate (6 g, 61.11 mmol), and palladium acetate (457 mg, 2.04) were sequentially added to DMSO (100 mL). Methyl), tricyclohexylphosphine (950 mg, 3.05 mmol), warmed to 100 ° C, and stirred at room temperature for 3 h. The mixture was cooled to room temperature, poured into water (300 mL), ethyl acetate (100 mL×3), washed with water (200mL×3), dried over anhydrous sodium sulfate, filtered, concentrated and evaporated The rate is 83.4%. LC-MS (APCI): m / z = 319.2 (M + 1) +.

Step 2 Synthesis of Compound 97.

Under a nitrogen atmosphere, compound 96 (2 g, 6.29 mmol), compound 2 (1.3 g, 6.29 mmol), sodium carbonate (2 g, 18.87 mmol), tetratriphenylphosphine palladium, and acetonitrile/water (20 mL/10 mL) were sequentially added. (400 mg, 0.173 mmol), warmed to reflux, and stirred at room temperature for 3 h. After cooling to room temperature, the insoluble material was filtered off, the acetonitrile was evaporated, ethyl acetate was evaporated (EtOAc, EtOAc (EtOAc) 75.0%. LC-MS (APCI): m / z = 361.0 (M + 1) +.

Step 3 Synthesis of Compound 99.

Compound 97 (322 mg, 0.896 mmol), Compound 98 (200 mg, 0.746 mmol), cesium carbonate (608 mg, 1.865 mmol), Pd(OAc) ₂ (18 mg, 0.075 mmol) were added to DMF (20 mL). Xantphos (86 mg, 0.149 mmol) was warmed to 105 ° C and stirred to react overnight. After cooling to room temperature, ethyl acetate (60 mL) was added, and the insoluble solid was filtered, and concentrated, and the residue was passed through silica gel column to yield a white solid (200 mg, yield: 45%). LC-MS (APCI): m / z = 593.2 (M + 1) +.

Step 4 Synthesis of Compound 100.

Compound 99 (200 mg, 0.338 mmol), iron powder (113 mg, 2.03 mmol) and ammonium chloride (35 mg, 0.67 mmol) were sequentially added to a mixture of ethanol/water (6 mL / 2 mL) under a nitrogen atmosphere, and heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and the organic phase was collected to give 189 mg of solid product, yield 99.0%. LC-MS (APCI): m / z = 563.2 (M + 1) +.

Step 5 Synthesis of Compound T-21.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 5 mL of dichloromethane was sequentially added with compound 100 (189 mg, 0.338 mol). After it was completely dissolved, triethylamine (69 mg, 686 umol) was added dropwise to the reaction system. After cooling to -10 ° C, compound 9 (30 mg, 0.338 mmol) was added slowly and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography The yield was 38.4%. LC-MS (APCI): m/z = 617.2 (M+1). 1H NMR (500MHz, CDCl3) (δ/ppm): 9.79 (s, 1H), 9.58 (br s, 1H), 8.15 (s, 1H), 7.96 (d, J = 11.5 Hz, 1H), 7.91 (d, J = 5.5 Hz, 1H), 7.75 (s, 1H), 7.49 (d, J = 5.5 Hz, 1H), 6.74 (s, 1H), 6.48 (dd, J = 17.0 Hz, J = 1.5 Hz, 1H) , 5.71 (d, J = 10.0 Hz, 1H), 4.78 - 4.74 (m, 1H), 3.90 (s, 3H), 3.16 - 3.09 (m, 2H), 2.74 - 2.72 (m, 2H), 2.71 (s , 3H), 2.69 (s, 3H), 2.56 (s, 6H), 1.72 (s, 3H), 1.70 (s, 3H).

Example 22 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-(4-(1-methyl-1-H-indole-5-)thiophene And [2,3-d] Pyrimidine-2-amino)acrylamide (T-22)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 102.

Under a nitrogen atmosphere, a solution of compound 2 (1.9 g, 9.22 mmol) and compound 101 (1.5 g, 8.38 mmol) in acetonitrile / water (2 / 1) (45 mL) was added Na ₂ CO ₃ (1.8 g, 16.76 mmol) ), Pd(pph ₃ ) ₄ (0.484 g, 0.42 mmol), heated to reflux, and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was evaporated, evaporated, evaporated, evaporated, evaporated LC-MS (APCI): m / z = 300.2 (M + 1) +.

Step 2 Synthesis of Compound 103.

Compound 102 (1.0 g, 3.34 mmol), compound 4 (0.62 g, 3.34 mmol), dissolved in 2-pentanol (8 mL), and added p-toluenesulfonic acid monohydrate (0.76 g, 4.0 mmol). The reaction mixture was reacted at 105 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, aq. EtOAc EtOAc (EtOAc m. %. LC-MS (APCI): m / z = 450.1 (M + 1) +.

Step 3 Synthesis of Compound 104.

Compound 103 (780 mg, 1.79 mmol), compound 6 (219 mg, 2.15 mmol), potassium carbonate (495 mg, 3.58 mmol) and DMF (5 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and filtered, and then filtered, and the organic phase was collected and purified by column chromatography to afford 560 mg of product as an orange solid. LC-MS (APCI): m / z = 532.2 (M + 1) +. 1H NMR (400MHz, CDCl3) (δ / ppm) 9.60 (s, 1H), 8.22 (dd, J = 8.4 Hz, J = 1.6 Hz, 1H), 7.93 (d, J = 5.6 Hz, 1H), 7.71 (s) , 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.40 (d, J = 1.6 Hz, 1H), 7.15 (d, J = 3.2 Hz, 1H), 6.71-6.69 (m, 2H), 4.00 (s, 3H), 3.88 (s, 3H), 3.28 (t, J = 7.2 Hz, 2H), 2.90 (s, 3H), 2.57 (t, J = 7.2 Hz, 2H), 2.27 (s, 6H) .

Step 4 Synthesis of Compound 105.

Compound 104 (560 mg, 1.06 mmol), iron powder (354 mg, 6.3 mmol) and ammonium chloride (226 mg, 4.22 mmol) were added to a mixture of ethanol/water (15 mL/5 mL) in a nitrogen atmosphere, and then heated. After refluxing and reacting for 1 hour, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and the organic phase was collected to give 530 mg of solid product, yield 99.9%. LC-MS (APCI): m / z = 502.1 (M + 1) +.

Step 5 Synthesis of Compound T-22.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 10 mL of dichloromethane was added with a compound 105 (530 mg, 1.06 mmol), and after it was completely dissolved, triethylamine (120 mg, 1.16 mmol) was added dropwise. The system was cooled to -10 ° C. Compound 9 (100 mg, 1.16 mol) was slowly added and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 200 mg, the yield was 34.0%. LC-MS (APCI): m / z = 556.2 (M + 1) +. 1H NMR (500MHz, CDCl3) (δ/ppm) 10.04 (br s, 1H), 9.91 (s, 1H), 8.67 (s, 1H), 8.25 (d, J = 7.5 Hz, 1H), 7.88 (d, J) =5.0 Hz, 1H), 7.80 (s, 1H), 7.49 (t, J = 7.0 Hz, 2H), 7.12 (d, J = 3.0 Hz, 1H), 6.80 (s, 1H), 6.68 (dd, J =3.0 Hz, J=1.0 Hz, 1H), 6.52-6.48 (m, 1H), 6.39-6.35 (m, 1H), 5.70 (dd, J = 10.5 Hz, J = 1.5 Hz, 1H), 3.90 (s) , 3H), 3.86 (s, 3H), 2.90 (t, J = 5.5 Hz, 2H), 2.71 (s, 3H), 2.29 (t, J = 5.5 Hz, 2H), 2.26 (s, 6H).

Example 23 N-(2-{2-Dimethylaminoethyl-methylamino}-5-(4-(4-fluoro-1-isopropyl-2-methyl-1Hbenzo[d]imidazole -6-) thiophene Benzo[2,3-d]pyrimidin-2-amino)-4-methoxyphenyl)acrylamide (T-23)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 106.

Under a nitrogen atmosphere, compound 96 (2 g, 6.29 mmol), compound 95 (1.3 g, 6.29 mmol), sodium carbonate (2 g, 18.87 mmol), tetratriphenylphosphine palladium, and the like, were sequentially added to acetonitrile/water (20 mL / 10 mL). (400 mg, 0.173 mmol), warmed to reflux, and stirred at room temperature for 3 h. After cooling to room temperature, the insoluble material was filtered off, the acetonitrile was evaporated, ethyl acetate was evaporated (EtOAc, EtOAc (EtOAc) 75.0%. LC-MS (APCI): m / z = 361.0 (M + 1) +.

Step 3 Synthesis of Compound 107.

Compound 97 (322 mg, 0.896 mmol), Compound 98 (200 mg, 0.746 mmol), cesium carbonate (608 mg, 1.865 mmol), Pd(OAc) ₂ (18 mg, 0.075 mmol) were added to DMF (20 mL). Xantphos (86 mg, 0.149 mmol) was warmed to 105 ° C and stirred to react overnight. After cooling to room temperature, ethyl acetate (60 mL) was added, and the insoluble solid was filtered, and concentrated, and the residue was passed through silica gel column to yield a white solid (200 mg, yield: 45%). LC-MS (APCI): m / z = 593.2 (M + 1) +.

Step 4 Synthesis of Compound 108.

Compound 99 (200 mg, 0.338 mmol), iron powder (113 mg, 2.03 mmol) and ammonium chloride (35 mg, 0.67 mmol) were sequentially added to a mixture of ethanol/water (6 mL / 2 mL) under a nitrogen atmosphere, and heated. After refluxing and reacting for 2 hours, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and the organic phase was collected to give 189 mg of solid product, yield 99.0%. LC-MS (APCI): m / z = 563.2 (M + 1) +.

Step 5 Synthesis of Compound T-23.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 5 mL of dichloromethane was sequentially added with compound 100 (189 mg, 0.338 mol). After it was completely dissolved, triethylamine (69 mg, 686 umol) was added dropwise to the reaction system. After cooling to -10 ° C, compound 9 (30 mg, 0.338 mmol) was added slowly and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography The yield was 38.4%. LC-MS (APCI): m / z = 617.2 (M + 1) +. ¹ H NMR (300 MHz, CDCl ₃ ) (δ / ppm) 10.08 (br s, 1H), 9.76 (s, 1H), 7.92 (d, J = 1.2 Hz, 1H), 7.77 (s, 1H), 7.72 ( d, J = 11.8 Hz, 1H), 7.42 (d, J = 6.0 Hz, 1H), 7.19 (d, J = 6.0 Hz, 1H), 6.81 (s, 1H), 6.52-6.45 (m, 1H), 6.39-6.36 (m, 1H), 5.73-5.69 (m, 1H), 4.83-4.76 (m, 1H), 3.90 (s, 3H), 2.90 (t, J = 6.0 Hz, 2H), 2.72 (s, 3H), 2.71 (s, 3H), 2.32 (t, J = 6.0 Hz, 2H), 2.29 (s, 6H), 1.70 (s, 3H), 1.67 (s, 3H).

Example 24 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-(4-(1-methyl-1-H-indole-5-)thiophene And [2,3-d] Pyrimidine-2-amino)acrylamide (T-24)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 110.

Under a nitrogen atmosphere, a solution of compound 95 (1.9 g, 9.22 mmol) and compound 101 (1.5 g, 8.38 mmol) in acetonitrile / water (2 / 1) (45 mL) was added Na ₂ CO ₃ (1.8 g, 16.76 mmol) ), Pd(pph ₃ ) ₄ (0.484 g, 0.42 mmol), heated to reflux, and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was evaporated, evaporated, evaporated, evaporated, evaporated LC-MS (APCI): m / z = 300.2 (M + 1) +.

Step 2 Synthesis of Compound 111.

Under the protection of nitrogen, compound 110 (1.0 g, 3.34 mmol), compound 4 (0.62 g, 3.34 mmol) was dissolved in 2-pentanol (8 mL), and p-toluenesulfonic acid monohydrate (0.76 g, 4.0 mmol) was added. The reaction mixture was reacted at 105 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, aq. EtOAc EtOAc (EtOAc m. %. LC-MS (APCI): m / z = 450.1 (M + 1) +.

Step 3 Synthesis of Compound 112.

Compound 11 (780 mg, 1.79 mmol), compound 6 (219 mg, 2.15 mmol), potassium carbonate (495 mg, 3.58 mmol) and DMF (5 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and filtered, and then filtered, and the organic phase was collected and purified by column chromatography to afford 560 mg of product as an orange solid. LC-MS (APCI): m / z = 532.2 (M + 1) +.

Step 4 Synthesis of Compound 113.

Compound 104 (560 mg, 1.06 mmol), iron powder (354 mg, 6.3 mmol) and ammonium chloride (226 mg, 4.22 mmol) were added to a mixture of ethanol/water (15 mL/5 mL) in a nitrogen atmosphere, and then heated. After refluxing and reacting for 1 hour, it was cooled to room temperature, ethanol was removed, water was added, and the mixture was extracted with dichloromethane, and the organic phase was collected to give 530 mg of solid product, yield 99.9%. LC-MS (APCI): m / z = 502.1 (M + 1) +.

Step 5 Synthesis of Compound T-24.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 10 mL of dichloromethane was added with a compound 105 (530 mg, 1.06 mmol), and after it was completely dissolved, triethylamine (120 mg, 1.16 mmol) was added dropwise. The system was cooled to -10 ° C. Compound 9 (100 mg, 1.16 mol) was slowly added and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 200 mg, the yield was 34.0%. LC-MS (APCI): m / z = 556.2 (M + 1) +. ^{1 H NMR (500MHz, DMSO-} D6) (δ / ppm) 9.92 (br s, 1H), 8.63 (s, 1H), 8.46 (s, 1H), 8.40 (d, J = 7.5Hz, 1H), 8.30 (s, 1H), 7.78 (d, J = 6.5 Hz, 1H), 7.52 (d, J = 8.0 Hz, 1H), 7.44 (d, J = 6.5 Hz, 1H), 7.24 (t, J = 7.0 Hz) , 1H), 7.11 (t, J = 7.0 Hz, 1H), 7.01 (s, 1H), 6.78-6.67 (m, 1H), 6.23 (d, J = 16.5 Hz, 1H), 5.73 (d, J = 16.5 Hz, 1H), 3.92 (d, 3H), 2.83 (s, 3H), 3.34-3.32 (m, 2H), 3.21-3.18 (m, 2H), 2.67 (s, 3H), 2.52 (s, 6H) ).

Example 25 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-(4-(1-H-indol-5-)thieno[2, 3-d]pyrimidin-2- Amino)acrylamide (T-25)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 115.

Under a nitrogen atmosphere, a solution of compound 95 (1.9 g, 9.22 mmol) and compound 114 (2.2 g, 8.38 mmol) in acetonitrile / water (2 / 1) (45 mL) was added Na ₂ CO ₃ (1.8 g, 16.76 mmol) ), Pd(pph ₃ ) ₄ (0.484 g, 0.42 mmol), heated to reflux, and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was evaporated, evaporated, evaporated, evaporated. LC-MS (APCI): m / z = 386.1 (M + 1) +.

Step 2 Synthesis of Compound 116.

To a solution of Compound 116 (410 mg, 1.06 mmol) in DCM (10 mL). Methylene chloride and excess trifluoroacetic acid were spun off, ethyl acetate (30 mL) was added and the mixture was stirred for half an hour. The resulting solid was filtered, washed with ethyl acetate and dried to give 265 mg of a yellow solid. LC-MS (APCI): m / z = 286.1 (M + 1) +.

Step 3 Synthesis of Compound 117.

Compound 116 (266 mg, 0.886 mmol), compound 4 (165 mg, 0.886 mmol), dissolved in 2-pentanol (5 mL), and then added p-toluenesulfonic acid monohydrate (381 g, 2.22 mmol) was reacted under nitrogen. The mixture was reacted at 105 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, aq. EtOAc EtOAc (EtOAc m. %. LC-MS (APCI): m / z = 436.2 (M + 1) +.

Step 4 Synthesis of Compound 118.

Compound 117 (230 mg, 0.53 mmol), compound 6 (81 mg, 0.79 mmol), potassium carbonate (109 mg, 0.79 mmol) and DMF (3 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water and then filtered, and then filtered, and the organic phase was collected and purified by column chromatography to give 136 mg of the product as an orange solid. LC-MS (APCI): m / z = 518.2 (M + 1) +.

Step 5 Synthesis of Compound 119.

Under a nitrogen atmosphere, compound 118 (136 mg, 0.263 mmol), iron powder (88 mg, 1.582 mmol) and ammonium chloride (7 mg, 0.132 mmol) were added to a mixture of ethanol/water (6 mL / 2 mL) and heated. After refluxing and reacting for 1 hour, it was cooled to room temperature, ethanol was removed, water was added, and extracted with dichloromethane, and the organic phase was collected to give a solid product (128 mg, yield: 99.9%). LC-MS (APCI): m / z = 488.1 (M + 1) +.

Step 6 Synthesis of Compound T-25.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 10 mL of dichloromethane was added sequentially to compound 119 (128 mg, 0.263 mmol). After it was completely dissolved, triethylamine (30 mg, 0.29 mmol) was added dropwise. The system was cooled to -10 ° C, and the compound 9 (23 mg, 0.263 mol) was slowly added and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 40 mg, the yield was 28.0%. LC-MS (APCI): m/z = 542.2 (M+1 ⁾ . ^{1 H NMR (500MHz, DMSO-} D6) (δ / ppm) 11.90 (s, 1H), 10.08 (s, 1H), 8.74 (s, 1H), 8.36 (d, J = 2.5Hz, 1H), 8.35 ( d, J = 8.0 Hz, 1H), 8.30 (s, 1H), 7.73 (d, J = 6.0 Hz, 1H), 7.44 (d, J = 7.5 Hz, 1H), 7.90 (d, J = 5.5 Hz, 1H), 7.15 (t, J = 8.0 Hz, 1H), 7.03-7.01 (m, 2H), 6.43-6.37 (m, 1H), 6.21-6.16 (m, 1H), 5.73-5.71 (m, 1H) , 3.79 (s, 3H), 2.92-2.88 (m, 2H), 2.72 (s, 3H), 2.36-2.32 (m, 2H), 2.22 (s, 6H).

Example 26 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-(4-(1-H-indol-5-)thieno[3, 2-d]pyrimidine-2- Amino)acrylamide (T-26)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 120.

Under a nitrogen atmosphere, a solution of compound 2 (1.9 g, 9.22 mmol) and compound 114 (2.2 g, 8.38 mmol) in acetonitrile / water (2 / 1) (45 mL) was added Na ₂ CO ₃ (1.8 g, 16.76 mmol) ), Pd(pph ₃ ) ₄ (0.484 g, 0.42 mmol), heated to reflux, and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was evaporated, evaporated, evaporated, evaporated, evaporated LC-MS (APCI): m / z = 386.1 (M + 1) +.

Step 2 Synthesis of Compound 121.

To a solution of Compound 120 (410 mg, 1.06 mmol) in DCM (10 mL) Methylene chloride and excess trifluoroacetic acid were spun off, ethyl acetate (30 mL) was added and the mixture was stirred for half an hour. The resulting solid was filtered, washed with ethyl acetate and dried to give 265 mg of a yellow solid. LC-MS (APCI): m / z = 286.1 (M + 1) +.

Step 3 Synthesis of Compound 122.

Compound 121 (266 mg, 0.886 mmol), Compound 4 (165 mg, 0.886 mmol), dissolved in 2-pentanol (5 mL), and then added p-toluenesulfonic acid monohydrate (381 g, 2.22 mmol) was reacted under nitrogen. The mixture was reacted at 105 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, aq. EtOAc EtOAc (EtOAc m. %. LC-MS (APCI): m / z = 436.2 (M + 1) +.

Step 4 Synthesis of Compound 123.

Under a nitrogen atmosphere, compound 122 (230 mg, 0.53 mmol), compound 6 (81 mg, 0.79 mmol), potassium carbonate (109 mg, 0.79 mmol) and DMF (3 mL) were added to a three-neck round bottom flask and stirred at room temperature. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and filtered, and then filtered, and the organic phase was collected and purified by column chromatography to yield 136 mg of the product as an orange solid. LC-MS (APCI): m / z = 518.2 (M + 1) +.

Step 5 Synthesis of Compound 124.

Under a nitrogen atmosphere, compound 123 (136 mg, 0.263 mmol), iron powder (88 mg, 1.582 mmol) and ammonium chloride (7 mg, 0.132 mmol) were added to a mixture of ethanol/water (6 mL / 2 mL) and heated. After refluxing and reacting for 1 hour, it was cooled to room temperature, ethanol was removed, water was added, and extracted with dichloromethane, and the organic phase was collected to give a solid product (128 mg, yield: 99.9%). LC-MS (APCI): m / z = 488.1 (M + 1) +.

Step 6 Synthesis of Compound T-26.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 10 mL of dichloromethane was added sequentially to compound 119 (128 mg, 0.263 mmol). After it was completely dissolved, triethylamine (30 mg, 0.29 mmol) was added dropwise. The system was cooled to -10 ° C, and the compound 9 (23 mg, 0.263 mol) was slowly added and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 40 mg, the yield was 28.0%. LC-MS (APCI): m / z = 542.2 (M + 1) +. ^{1 H NMR (500MHz, CDCl3)} (δ / ppm): 9.91 (s, 1H), 8.67 (s, 1H), 8.25 (d, J = 7.5Hz, 1H), 7.88 (d, J = 5.0Hz, 1H ), 7.80 (s, 1H), 7.49 (t, J = 7.0 Hz, 2H), 7.12 (d, J = 3.0 Hz, 1H), 6.80 (s, 1H), 6.68 (dd, J = 3.0 Hz, J =1.0 Hz, 1H), 6.52-6.48 (m, 1H), 6.39-6.35 (m, 1H), 5.70 (dd, J = 10.5 Hz, J = 1.5 Hz, 1H), 3.90 (s, 3H), 3.86 (s, 3H), 2.90 (t, J = 5.5 Hz, 2H), 2.71 (s, 3H), 2.29 (t, J = 5.5 Hz, 2H), 2.26 (s, 6H).

Example 27 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-(4-(1-H-indazole)-3-)thieno[3, 2-d]pyrimidine-2- Amino)acrylamide (T-27)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 127.

Under a nitrogen atmosphere, compound 125 (2.4 g, 10 mmol) was dissolved in anhydrous THF (30 mL), cooled to 0 ° C, NaH (60%, 0.52 g, 13 mmol) was added, and the reaction was stirred at 0 ° C for 20 minutes, slowly added dropwise. A solution of trityl chloride (2.78 g, 10 mmol) in THF (5 mL). Ethyl acetate (60 mL) was added, and the obtained solid was filtered, and the filtrate was concentrated. LC-MS (APCI): m / z = 487.1 (M + 1) +.

Step 2 Synthesis of Compound 128.

Under a nitrogen atmosphere, compound 127 (0.97 g, 2.0 mmol), bispinacol borate (1.03 g, 4.0 mmol), potassium acetate (0.588 g, 6.0 mmol), and potassium acetate (0.588 g, 6.0 mmol) were added to dioxane (5 mL). Pd(dppf)Cl ₂ -DCM (81 mg, 0.1 mmol), warmed to 100 ° C, and stirred at room temperature overnight. After cooling to room temperature, ethyl acetate (30 mL) was added, filtered, evaporated and evaporated. LC-MS (APCI): m / z = 487.1 (M + 1) +.

Step 3 Synthesis of Compound 129.

To a solution of compound 128 (0.48 g, 1.0 mmol) and compound 2 (0.2 g, 1.0 mmol) in acetonitrile / water (2 / 1) (10 mL), Na ₂ CO ₃ (0.2 g, 2 mmol) , Pd(PPh ₃ ) ₄ (57 mg, 0.05 mmol), heated to reflux, and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was evaporated, evaporated, evaporated, evaporated, evaporated LC-MS (APCI): m / z = 529.2 (M + 1) +.

Step 4 Synthesis of Compound 130.

Compound 19 (350 mg, 0.663 mmol), compound 4 (120 mg, 0.663 mmol), dissolved in 2-pentanol (5 mL), and then added p-toluenesulfonic acid monohydrate (250 mg, 1.3 mmol). The mixture was reacted at 105 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, aq. EtOAc (EtOAc (EtOAc) (EtOAc) %. LC-MS (APCI): m / z = 679.2 (M + 1) +.

Step 5 Synthesis of Compound 131.

Compound 122 (300 mg, 0.44 mmol), compound 6 (81 mg, 0.79 mmol), potassium carbonate (109 mg, 0.79 mmol) and DMF (3 mL) were added to a three-neck round bottom flask and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and filtered, and then filtered, and the organic phase was collected and purified by column chromatography to give an orange solid product (240 mg, yield: 71.8%). LC-MS (APCI): m / z = 761.3 (M + 1) +. ¹ H NMR (400 MHz, CDCl 3 ) (δ / ppm) 9.27 (s, 1H), 8.01 (d, J = 8.0 Hz, 1H), 7.81 (d, J = 5.6 Hz, 1H), 7.63 (s, 1H) , 7.32-7.27 (m, 17H), 7.10 (t, J = 8.0 Hz, 1H), 6.75 (s, 1H), 6.63 (d, J = 8.0 Hz, 1H), 4.03 (s, 3H), 3.31 ( t, J = 6.8 Hz, 2H), 2.92 (s, 3H), 2.62 (t, J = 6.8 Hz, 2H), 2.31 (s, 6H).

Step 6 Synthesis of Compound 132.

Compound 131 (240 mg, 0.316 mmol), iron powder (114 mg, 1.89 mmol) and ammonium chloride (33 mg, 0.62 mmo) were sequentially added to a mixture of ethanol/water (6 mL/2 mL) under a nitrogen atmosphere, and heated. After refluxing and reacting for 1 hour, it was cooled to room temperature, ethanol was removed, water was added, extracted with dichloromethane, and the organic phase was collected to give a solid product (230 mg, yield: 99.9%). LC-MS (APCI): m / z = 731.1 (M + 1) +.

Step 7 Synthesis of Compound 133.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 10 mL of dichloromethane was added to the compound 132 (230 mg, 0.316 mmol). After it was completely dissolved, triethylamine (33 mg, 0.32 mmol) was added dropwise. The system was cooled to -10 ° C, and compound 9 (29 mg, 0.32 mol) was added slowly. Stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 100 mg, the yield was 40.40%. LC-MS (APCI): m / z = 785.1 (M + 1) +.

Step 8 Synthesis of Compound T-27.

Triethylsilane (0.29 g, 2.54 mmol) and trifluoroacetic acid (1.0 g, 8.89 mmol) were added dropwise to a solution of Compound 133 (100 mg, 0.127 mmol) in dichloromethane (3 mL). The reaction was stirred for 2 h. Saturated NaHCO ₃ (20mL) The reaction was quenched, (30mL x 3) and extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated and the residue was subjected to silica gel column to give a white solid 30mg, yield 43.6%. LC-MS (APCI): m / z = 543.2 (M + 1) +. ^{1 H NMR (400MHz, CDCl3)} (δ / ppm): 11.70 (s, 1H), 10.19 (s, 1H), 9.76-9.71 (m, 1H), 8.72 (d, J = 8.0Hz, 1H), 7.75 -7.73(m,1H), 7.65(s,1H),7.34-7.19(m,4H), 6.85-6.84(m,1H),6.52-6.51(m,1H),5.80-5.77(m,1H) , 3.94 (s, 3H), 3.31-3.29 (m, 2H), 2.77 (s, 3H), 2.57-2.55 (m, 2H), 2.42 (s, 6H).

Example 28 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-(4-(1-methyl-1-H-indazole)-3-) Thio[3,2-d] Pyrimidine-2-amino)acrylamide (T-28)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 134.

Triethylsilane (1.16 g, 10 mmol) and trifluoroacetic acid (3.42 g, 30 mmol) were added dropwise to a solution of Compound 129 (528 mg, 1.0 mmol) in dichloromethane (5 mL). 2h. Saturated NaHCO ₃ (20mL) The reaction was quenched, (30mL x 3) and extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated and the residue was subjected to silica gel column to give a white solid 200mg, yield 69.9%. LC-MS (APCI): m / z = 287.1 (M + 1) +.

Step 2 Synthesis of Compound 135.

The compound 134 (200 mg, 0.699 mmol) was dissolved in anhydrous THF (8 mL), cooled to 0 ° C, NaH (60%, 36 mg, 0.91 mmol), and the mixture was stirred at 0 ° C for 20 minutes, slowly added dropwise. A solution of methyl p-toluenesulfonate (0.17 g, 0.91 mmol) in THF (1 mL) Ethyl acetate (60 mL) was added, and the obtained solid was filtered, and the filtrate was concentrated. LC-MS (APCI): m / z = 301.1 (M + 1) +.

Step 3 Synthesis of Compound 136.

Under the protection of nitrogen, the compound 135 (180 mg, 0.60 mmol), the compound 4 (120 mg, 0.663 mmol) was dissolved in 2-pentanol (5 mL), and p-toluenesulfonic acid monohydrate (250 mg, 1.3 mmol) was added. The mixture was reacted at 105 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, aq. EtOAc EtOAc (EtOAc m. %. LC-MS (APCI): m / z = 451.1 (M + 1) +.

Step 4 Synthesis of Compound 137.

Compound 136 (130 mg, 0.29 mmol), compound 6 (81 mg, 0.79 mmol), potassium carbonate (109 mg, 0.79 mmol) and DMF (3 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and then filtered, and then filtered, and the organic phase was collected and purified by column chromatography to give an orange solid product (yield: 77.8%). LC-MS (APCI): m / z = 533.2 (M + 1) +.

Step 5 Synthesis of Compound 138.

Compound 131 (120 mg, 0.226 mmol), iron powder (114 mg, 1.89 mmol) and ammonium chloride (33 mg, 0.62 mmol) were added to a mixture of ethanol/water (6 mL / 2 mL) in a mixture. After refluxing and reacting for 1 hour, it was cooled to room temperature, ethanol was removed, water was added, extracted with dichloromethane, and the organic phase was collected to give a solid product (100 mg, yield: 88.1%). LC-MS (APCI): m / z = 503.1 (M + 1) +.

Step 6 Synthesis of Compound T-28.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 10 mL of dichloromethane was added to the compound 138 (100 mg, 0.20 mmol). After it was completely dissolved, triethylamine (23 mg, 0.22 mmol) was added dropwise. The system was cooled to -10 ° C, and the compound 9 (19 mg, 0.20 mol) was slowly added and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 40 mg, the yield was 36.0%. LC-MS (APCI): m / z = 557.1 (M + 1) +. ^{1 H NMR (400MHz, DMSO-} D6) (δ / ppm): 10.24 (br s, 1H), 9.56 (br s, 1H), 8.57-8.53 (m, 2H), 8.39 (s, 1H), 8.29 ( d, J = 4.8 Hz, 1H), 7.76 (d, J = 6.8 Hz, 1H), 7.48 (t, J = 6.0 Hz, 1H), 7.30-7.26 (m, 2H), 7.00 (s, 1H), 6.22-6.18(m,1H), 5.71-05.69(m,1H), 4.23(s,3H),3.83(s,3H),3.33-3.31(m,2H),2.77(s,3H),2.57- 2.55 (m, 2H), 2.42 (s, 6H).

Example 29 N-(2-{2-Dimethylaminoethyl-methylamino}-4-methoxy-5-(4-(1-H-吲哚)-3-)thieno[2, 2-d]pyrimidine-2- Amino)acrylamide (T-25)

The specific synthesis steps are as follows:

Step 1 Synthesis of Compound 139.

Under a nitrogen atmosphere, a solution of compound 2 (1.9 g, 9.22 mmol) and compound 114 (2.2 g, 8.38 mmol) in acetonitrile / water (2 / 1) (45 mL) was added Na ₂ CO ₃ (1.8 g, 16.76 mmol) ), Pd(pph ₃ ) ₄ (0.484 g, 0.42 mmol), heated to reflux, and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was evaporated, evaporated, evaporated, evaporated, evaporated LC-MS (APCI): m / z = 386.1 (M + 1) +.

Step 2 Synthesis of Compound 140.

To a solution of the compound 139 (410 mg, 1.06 mmol) in DCM (10 mL), trifluoroacetic acid (10 mL) was added dropwise, and the mixture was stirred at room temperature for 2 hours. Methylene chloride and excess trifluoroacetic acid were spun off, ethyl acetate (30 mL) was added and the mixture was stirred for half an hour. The resulting solid was filtered, washed with ethyl acetate and dried to give 265 mg of a yellow solid. LC-MS (APCI): m / z = 286.1 (M + 1) +.

Step 3 Synthesis of Compound 141.

Under the protection of nitrogen, compound 140 (266 mg, 0.886 mmol), compound 4 (165 mg, 0.886 mmol), and then dissolved in 2-pentanol (5 mL) were added, and p-toluenesulfonic acid monohydrate (381 g, 2.22 mmol) was reacted. The mixture was reacted at 105 ° C overnight under a liquid nitrogen atmosphere. After cooling to room temperature, aq. EtOAc EtOAc (EtOAc m. %. LC-MS (APCI): m / z = 436.2 (M + 1) +.

Step 4 Synthesis of Compound 142.

Compound 141 (230 mg, 0.53 mmol), compound 6 (81 mg, 0.79 mmol), potassium carbonate (109 mg, 0.79 mmol) and DMF (3 mL) were added to a three-neck round bottom flask, and stirred at room temperature under nitrogen. After 16 hours, it was diluted with 20 mL of water, filtered, washed with water, and filtered, and then filtered, and the organic phase was collected and purified by column chromatography to yield 136 mg of the product as an orange solid. LC-MS (APCI): m / z = 518.2 (M + 1) +.

Step 5 Synthesis of Compound 143.

Compound 142 (136 mg, 0.263 mmol), iron powder (88 mg, 1.582 mmol) and ammonium chloride (7 mg, 0.132 mmol) were sequentially added to a mixture of ethanol/water (6 mL / 2 mL) under a nitrogen atmosphere, and then heated. After refluxing and reacting for 1 hour, it was cooled to room temperature, ethanol was removed, water was added, and extracted with dichloromethane, and the organic phase was collected to give a solid product (128 mg, yield: 99.9%). LC-MS (APCI): m / z = 488.1 (M + 1) +.

Step 6 Synthesis of Compound T-29.

Under a nitrogen atmosphere, a single-necked round bottom flask containing 10 mL of dichloromethane was added to the compound 143 (128 mg, 0.263 mmol). After it was completely dissolved, triethylamine (30 mg, 0.29 mmol) was added dropwise. The system was cooled to -10 ° C, and the compound 9 (23 mg, 0.263 mol) was slowly added and stirring was continued for 30 minutes. The reaction was quenched with saturated sodium hydrogen carbonate solution and extracted with dichloromethane. The organic phase was collected and purified by column chromatography to give a yellow solid product. 40 mg, the yield was 28.0%. LC-MS (APCI): m / z = 542.2 (M + 1) +. ^{1 H NMR (500MHz, DMSO-} D6) (δ / ppm): 11.90 (s, 1H), 10.08 (s, 1H), 8.74 (s, 1H), 8.47 (d, J = 2.5Hz, 1H), 8.29 -8.23(m,2H), 8.13(s,1H), 7.48(d,J=6.0Hz,1H), 7.30-7.28(m,1H), 7.20-7.17(m,1H),7.12-7.08(m , 1H), 7.00 (s, 1H), 6.67-6.61 (m, 1H), 6.22-6.18 (m, 1H), 5.73-5.69 (m, 1H), 3.79 (s, 3H), 2.92-2.88 (m , 2H), 2.72 (s, 3H), 2.36-2.32 (m, 2H), 2.22 (s, 6H).

Biological activity test

(1) Kinase inhibition experiment

Reagents and consumables:

Eu-anti-GST抗体(Life Technologies,目录号PV5594)。WT EGFR (Carna, Cat. No. 08-115), EGFR [L858R] (Carna, Cat. No. 08-502) EGFR [L858R/T790M] (Carna, Cat. No. 08-510), ATP (Sigma, Cat. No. A7699-1G) ), DMSO (Sigma, Cat. No. D2650), 96-well plate (Corning, Cat. No. 3365), 384-well plate (Greiner, Cat. No. 784076), HTRF Kinase TK Kit (Cisbio, Cat. No. 62TK0PEJ), Erlotinib (Selleckchem, Cat. No. S7787), EGFR [d746-750] (Life Technologies, Cat. No. PV6178), 5x Kinase Buffer A (Life Technologies, Cat. No. PV3186), Kinase Tracer 199 (Life Technologies, Cat. No. PV5830) ,

Eu-anti-GST antibody (Life Technologies, catalog number PV5594).

Specific experimental methods:

Compound formulation: The test compound was dissolved in DMSO to make a 20 mM mother liquor. Then, it was diluted 3 times in a medium gradient of DMSO and diluted 10 times. When dosing, dilute with buffer for 10 times.

WT EGFR, EGFR [L858R] and EGFR [L858R/T790M] kinase assays: WT EGFR, EGFR [L858R] or EGFR [L858R/T790M] kinases in pre-diluted compounds at 5x kinase buffer A Mix for 10 minutes, double wells per concentration. The corresponding substrate and ATP in the kit were added and reacted at room temperature for 20 minutes (where a positive control was set: negative for blank control and positive for erlotinib). After the reaction, the detection reagent (the reagent in the HTRF Kinase TK kit) was added, and after incubation at room temperature for 30 minutes, the enzyme activity in the presence of the compound of the present invention at each concentration was measured by an Evnvision microplate reader, and the compound concentrations of different concentrations were calculated. The inhibitory activity of the enzyme activity, and then according to the four-parameter equation, according to Graphpad 5.0 software, the inhibitory activity of the enzyme activity under different concentrations of compounds was fitted, and the IC ₅₀ value was calculated, wherein A means IC ₅₀ ≤ 1 nM, B means IC ₅₀ It is 1-10 nM, C means that the IC ₅₀ is 10-50 nM, and D means that the IC ₅₀ is 50-100 nM.

The compounds of the present invention were tested in the above kinase inhibition assay, and the compounds of the present invention were found to have potent activity against EGFR [L858R] and EGFR [L858R/T790M] and superior selectivity to WT EGFR. The results for the example compounds are summarized in Table 1 below.

Table 1

(2) Cytotoxicity experiment

The anti-proliferative activity of the compound of the present invention against three tumor cells cultured in vitro was examined by MTS method. The experimental results show that the compound of the present invention has an inhibitory effect on the in vitro proliferation of cancer cells cultured in vitro; wherein the inhibition of proliferation of lung cancer cells in vitro is stronger than that of skin cancer cells in vitro.

Cell line:

Skin cancer cell line A431 (purchased from the American Standard Collection of Biological Products (ATCC)); lung cancer cells NCI-H1975 (purchased from the American Standard Collection of Biological Products (ATCC)) and HCC827 (purchased from the American Standard Collection of Biological Products (ATCC) Both were cultured in RPMI1640 medium containing 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin.

Reagents and consumables:

RPMI-1640 (GIBCO, catalog number A10491-01); fetal bovine serum (GIBCO, catalog number 10099141); 0.25% trypsin-EDTA (GIBCO, catalog number 25200); penicillin-streptomycin, liquid (GIBCO, catalog number 15140-122); DMSO (Sigma, Cat. No. D2650); MTS Test Kit (Promega, Cat. No. G3581), 96-well plate (Corning, Cat. No. 3365).

Specific experimental methods:

Compound preparation: The test compound was dissolved in DMSO to prepare a 20 mM mother liquor and stored at -20 °C. When used, it was diluted 3 times with a gradient of DMSO and diluted 10 times. The drug medium was further diluted 4 times with the cell culture medium RPMI-1640.

MTS cell viability assay: 0.25% trypsin-EDTA digested logarithmic growth phase cells, inoculated with 150 μl in 96-well plates at an optimized density, and added to the medium RPMI-1640 diluted 4 times after 24 hours, 50 μl/well (general Ten concentrations were selected: 100, 33.3, 11.1, 3.70, 1.23, 0.412, 0.137, 0.0457, 0.0152, 0.00508 μM). A well of the same volume of 0.5% DMSO was added as a control. After the cells were cultured for 72 hours, MTS was assayed for cell viability.

Specific procedure: adherent cells, discard the medium, and add a mixture containing 20 μL of MTS and 100 μL of medium per well. OD490 was detected after being placed in an incubator for 1-4 hours, with the OD650 value as a reference. Produced dose-response curve calculated by GraphPad Prism software and IC _50, wherein A means a IC ₅₀ ≤100nM, B means an IC ₅₀ of 100-500nM, C means an IC ₅₀ of 500-1000 nm, and D means an IC ₅₀ ≥1000nM .

The compounds of the invention were tested in the above cytotoxicity assay and found to be compounds of the invention against lung cancer cells NCI-H1975 and HCC827 has potent activity and superior selectivity to skin cancer cells A431. The results of the inhibition of in vitro proliferation of cancer cells by the examples are summarized in Table 2 below.

Table 2

(3) Metabolic stability experiment

Microsomal experiments: human liver microsomes: 0.5 mg/mL, BD Gentest; rat liver microsomes: 0.5 mg/mL, Xenotech; mouse liver microsomes: 0.5 mg/mL, Xenotech; coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

Preparation of stock solution: A certain amount of the compound powder of the example was accurately weighed and dissolved to 5 mM with DMSO.

Preparation of phosphate buffer (100 mM, pH 7.4): Mix 150 mL of pre-formed 0.5 M potassium dihydrogen phosphate and 700 mL of 0.5 M potassium dihydrogen phosphate solution, and adjust the mixture with 0.5 M potassium dihydrogen phosphate solution. The pH was adjusted to 7.4, diluted 5 times with ultrapure water before use, and magnesium chloride was added to obtain a phosphate buffer (100 mM) containing 100 mM potassium phosphate, 3.3 mM magnesium chloride, and a pH of 7.4.

A solution of NADPH regeneration system (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was prepared and placed on wet ice before use.

Formulation stop solution: acetonitrile solution containing 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard). Take 25057.5 μL of phosphate buffer (pH 7.4) into a 50 mL centrifuge tube, add 812.5 μL of human liver microsomes, and mix to obtain a liver microsome dilution with a protein concentration of 0.625 mg/mL. Take 25057.5 μL of phosphate buffer (pH 7.4) into a 50 mL centrifuge tube, add 812.5 μL of rat liver microsomes, and mix to obtain a liver microsome dilution with a protein concentration of 0.625 mg/mL. 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, and 812.5 μL of mouse liver microsomes were added and mixed to obtain a liver microsome dilution having a protein concentration of 0.625 mg/mL.

Incubation of the sample: The stock solution of the corresponding compound was diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile as a working solution, and was used. 398 μL of human liver microsomes or rat liver microsomes or mouse liver microsome dilutions were added to 96-well incubation plates (N=2), and 2 μL of 0.25 mM working solution was added and mixed.

Determination of metabolic stability: 300 μL of pre-cooled stop solution was added to each well of a 96-well deep well plate and placed on ice as a stop plate. The 96-well incubation plate and the NADPH regeneration system were placed in a 37 ° C water bath, shaken at 100 rpm, and pre-incubated for 5 min. 80 μL of the incubation solution was taken from each well of the incubation plate, added to the stopper plate, and mixed, and 20 μL of the NADPH regeneration system solution was added as a sample of 0 min. Then, 80 μL of the NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and start timing. The corresponding compound had a reaction concentration of 1 μM and a protein concentration of 0.5 mg/mL. 100 μL of the reaction solution was taken at 10, 30, and 90 min, respectively, and added to the stopper, and the reaction was terminated by vortexing for 3 min. The plate was centrifuged at 5000 x g for 10 min at 4 °C. 100 μL of the supernatant was taken into a 96-well plate to which 100 μL of distilled water was previously added, mixed, and sample analysis was performed by LC-MS/MS.

Data analysis: The peak area of the corresponding compound and the internal standard was detected by LC-MS/MS system, and the ratio of the peak area of the compound to the internal standard was calculated. The slope is measured by the natural logarithm of the percentage of the remaining amount of the compound versus time, and t _1/2 and CL _{int are} calculated according to the following formula, where V/M is equal to 1/protein concentration.

The compounds of the present invention were tested in the above microsomal experiments and found to have superior metabolic stability. The results of the human liver microsome experiment and the rat liver microsome experiment of the representative examples are summarized in Table 3 below.

table 3:

(4) Pharmacokinetic experiments in rats

Eight male Sprague-Dawley rats, 7-8 weeks old, weighing 210 g, were divided into 2 groups, 4 in each group, and a single oral dose of 5 mg/kg (a) control group: (R)-3-( 4-(2-(2-methyltetrazol-5-yl)pyridin-5-yl)-3-fluorophenyl)-5-hydroxymethyloxazolidin-2-one; (b) Test group: Example compounds were compared for differences in pharmacokinetics.

Rats were fed a standard diet and given water. Fasting began 16 hours before the test. The drug was dissolved with PEG400 and dimethyl sulfoxide. Blood was collected from the eyelids at a time point of 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours after administration.

Rats were briefly anesthetized after inhalation of ether, and 300 μL of blood samples were collected from the eyelids in test tubes. There was 30 μL of 1% heparin salt solution in the test tube. The tubes were dried overnight at 60 ° C before use. After the blood sample collection was completed at a later time point, the rats were anesthetized with ether and sacrificed.

Immediately after the blood sample is collected, gently invert the tube at least 5 times to ensure adequate mixing and place on ice. Blood samples were centrifuged at 5000 rpm for 5 minutes at 4 ° C to separate plasma from red blood cells. Pipette 100 μL of plasma into a clean plastic centrifuge tube, indicating the name and time of the compound. Plasma was stored at -80 °C prior to analysis. The concentration of the compound of the invention in plasma was determined by LC-MS/MS. Pharmacokinetic parameters were calculated based on the plasma concentration of each animal at different time points.

The compounds of the invention were tested in the above pharmacokinetic experiments in rats and found to have certain pharmacokinetic properties in animals. The results of the pharmacokinetic experiments of the representative Example 2 and the rat of Example 29 are summarized in Table 4 below.

Table 4:

It is to be understood that the examples are merely illustrative of the invention and are not intended to limit the scope of the invention, and the experimental methods in which the specific conditions are not indicated in the examples, usually in accordance with conventional conditions or in accordance with the conditions suggested by the manufacturer. Parts and percentages are parts by weight and percentage by weight unless otherwise stated.

The above is a further detailed description of the present invention in connection with the specific preferred embodiments, and the specific embodiments of the present invention are not limited to the description. It will be apparent to those skilled in the art that the present invention may be made without departing from the spirit and scope of the invention.

Claims (19)

Compound of formula (I):

among them, R ^{1 is} selected from H, halogen, -OH, -CN, -NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy base; L is selected from the group consisting of a bond, NR, O, CR ₂ or S; wherein R is independently selected from H, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; R ^{2 is} selected from a C ₁ -C ₆ alkyl group or a 3- to 6-membered heterocyclic group containing 1-2 hetero atoms selected from N and O, wherein the above group is unsubstituted or substituted by 1 to 3 substituents Substituents: halogen, -OH, -CN, -NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, NR ⁶ R ⁷ ; wherein R ⁶ , R ⁷ are each independently selected from C ₁ - C ₆ alkyl, C ₁ -C ₆ alkoxy or C ₃ -C ₆ carbocyclyl; R ³ , R ⁴ and R ^{5 are} independently selected from H, halogen, -CN, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl; Ring A is selected from a C ₃ -C ₆ carbocyclic group, a 3 to 10 membered heterocyclic group, a C ₆ -C ₁₄ aryl group or a C ₅ -C ₁₀ heteroaryl group; Ring B is selected from the following structures:

Wherein each X ^{1 is} independently selected from the group consisting of C, N, O and S atoms, each X ² and X ^{3 being} independently selected from C and N atoms, wherein the ring containing X ¹ , X ² and X ³ is optionally 1- 3 R ⁸ substituents substituted; Ring C is selected from a C ₃ -C ₆ carbocyclic group, a 3 to 10 membered heterocyclic group, a C ₆ -C ₁₄ aryl group or a C ₅ -C ₁₀ heteroaryl group, which is optionally substituted by 1 to 3 R ⁸ Base substitution

Represents a single or double bond; R ^{8 is} independently selected from H, halogen, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy; Or two R ⁸ on the ring together form -Z _m -; wherein Z is independently selected from CH ₂ , NH, O, S, SO or SO ₂ ; m = 2, 3, 4 or 5; Or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative thereof. The compound according to claim 1, wherein R ^{1 is} selected from the group consisting of -OH, halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkane O, OH, L is selected from NR, O or S, wherein R is selected from C ₁ -C ₆ alkyl, and R ³ , R ⁴ and R ⁵ are H. The compound according to claim 1 or 2, wherein R ¹ is a methoxy group, L is NR, wherein R is a methyl group, and R ³ , R ⁴ and R ⁵ are H. The compound according to any one of claims 1 to 3, wherein R ² is a C ₁ -C ₆ alkyl group substituted by 1 to 3 substituents, wherein the substituent is a C ₁ -C ₆ alkane Further, C ₁ -C ₆ alkoxy, NR ⁶ R ⁷ ; more preferably, R ² is CH ₂ CH ₂ N(CH ₃ ) ₂ . The compound according to any one of claims 1 to 4, wherein Ring A is selected from a C ₅ -C ₆ carbocyclic group, a 5- to 6-membered heterocyclic group, a C ₆ aryl group or a C ₅ -C ₆ heteroaryl group. More preferably, ring A is selected from a 5 to 6 membered heterocyclic group or a C ₅ -C ₆ heteroaryl group; more preferably, ring A is selected from a C ₅ -C ₆ heteroaryl group; most preferably, ring A is selected From C ₅ heteroaryl. The compound according to any one of claims 1 to 5, wherein ring A has the following structure:

among them, Each Y ^{1 is} independently selected from a C, N, O or S atom, each Y ^{2 is} independently selected from a C or N atom, and Y ¹ and Y ^{2 are} optionally substituted with an R ⁹ substituent selected from the group consisting of H, halogen, -CN, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy or C ₁ -C ₆ alkylsulfonyl, wherein C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy or C ₁ -C ₆ alkylsulfonyl optionally with one or more -OH substitution. The compound according to any one of claims 1 to 6, wherein ring A has the following structure:

Wherein R ⁹ is independently selected from H, C ₁ -C ₆ alkyl, substituted with one or more -OH, C ₁ -C ₆ alkyl. The compound according to any one of claims 1 to 7, wherein Ring B has the following structure:

Wherein each X ^{1 is} independently selected from the group consisting of C, N, O and S atoms, each X ² and X ^{3 being} independently selected from C and N atoms, wherein the ring containing X ¹ , X ² and X ³ is optionally 1- 3 R ⁸ substituents substituted; Ring C is selected from a C ₅ -C ₆ carbocyclic group, a 5 to 6 membered heterocyclic group, a C ₆ -C ₁₀ aryl group or a C ₅ -C ₆ heteroaryl group, which is optionally substituted with from 1 to 3 R ⁸ Base substitution

Represents a single or double bond; R ^{8 is} independently selected from H, halogen, oxo, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy; Or the two R ⁸ on the ring together form -(CH ₂ ) _m -, -NH(CH ₂ ) _n -, -O(CH ₂ ) _n -, -S(CH ₂ ) _n -, -SO(CH _{2 n} -, -SO ₂ (CH ₂ ) _n -, -CH ₂ NH(CH ₂ ) _p -, -CH ₂ O(CH ₂ ) _p -, -CH ₂ S(CH ₂ ) _p -, -CH ₂ SO(CH ₂ ) _p - or -CH ₂ SO ₂ (CH ₂ ) _p -; wherein m = 2, 3, 4 or 5; n = 1, 2 or 3; p = 1 or 2. The compound according to claim 8, wherein the ring C is selected from a C ₅ -C ₆ carbocyclic group, a 5- to 6-membered heterocyclic group, a C ₆ aryl group or a C ₅ -C ₆ heteroaryl group; more preferably, the ring C is selected from the group consisting of cyclopentadienyl, cyclohexadienyl, phenyl, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, dihydropyridyl, pyrimidinyl, pyrazinyl, dihydropyrazinyl. The compound according to any one of claims 8 or 9, wherein R ^{8 is} independently selected from H, F, oxo and C ₁ -C ₆ alkyl; or the two R ⁸ on the ring together form -(CH ₂ ) ₂ -, -(CH ₂ ) ₃ -, -OCH ₂ -, -O(CH ₂ ) ₂ - or -CH ₂ OCH ₂ -. The compound according to any one of claims 1 to 10, wherein Ring B has the following structure:

Wherein each ring is optionally substituted with from 1 to 3 R ⁸ substituents, and R ⁸ , X ¹ and X ^{2 are} as defined in any one of claims 1-10. The compound according to any one of claims 1 to 11, wherein the ring B is selected from the group consisting of:

Wherein R ^{8 is} as defined in any one of claims 1-10. A compound according to claim 1 which may be selected from the group consisting of:

A pharmaceutical composition comprising a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative thereof, And a pharmaceutically acceptable excipient. The pharmaceutical composition of claim 14 further comprising an additional therapeutic agent. Kit, which includes a first container comprising a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative thereof; Optionally, a second container containing other therapeutic agents; and Optionally, a third container containing a pharmaceutically acceptable excipient for diluting or suspending the compound and/or other therapeutic agent. A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, crystal form, prodrug or isotopic derivative thereof, for use in the treatment of cancer caused by EGFR Use of the drug. A method of treating cancer caused by EGFR in a subject, the method comprising administering to the subject a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, stereoisomerism A pharmaceutically, solvate, hydrate, crystal form, prodrug or isotope derivative, or a pharmaceutical composition according to any one of claims 14-15. The method of claim 17, or the method of 18, wherein the cancer caused by the EGFR is selected from the group consisting of: non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer , epithelial cell carcinoma, gastrointestinal stromal tumor, leukemia, histiocytic lymphoma, nasopharyngeal carcinoma.