CN103421010A - Pteridinone derivative as EGFR inhibitor and application thereof - Google Patents

Abstract

The present invention relates to pteridinone derivatives as EGFR inhibitors and their application. Specifically, the present invention relates to the compound represented by the following formula I, the pharmaceutical composition containing the compound of the following formula I and the use of the compound in the preparation of medicaments for treating EGFR-mediated diseases or inhibiting EGFR.

Description

Pteridinone derivatives as EGFR inhibitors and applications thereof

technical field

The present invention relates to the synthesis of pteridinone compounds and their application in the fields of medicinal chemistry and drug therapeutics, in particular to the application of pteridinone compounds with different substituents in the preparation of medicines for tumor-related diseases. The compound has high activity of inhibiting epidermal growth factor receptor tyrosine kinase (EGFR), and thus has the potential to treat tumors and related diseases.

Background technique

Malignant tumors are cellular lesions characterized by loss of control of normal cell division, leading to uncontrolled differentiation and proliferation, and can invade local tissues to cause metastasis. Malignant tumors have become a common disease that seriously endangers human life and health. According to incomplete statistics, there are nearly 20 million new cases every year in the world. Therefore, the research and development of anticancer drugs is a very challenging and significant field in today's life sciences.

Traditional antineoplastic drugs are mainly cytotoxic drugs, which have unavoidable side effects, poor selectivity, and easy drug resistance. With the rapid development of life science and technology, various basic life processes such as signal transduction, cell cycle regulation, and angiogenesis in malignant tumor cells are gradually being elucidated. Taking certain key enzymes of signal transduction pathways related to tumor cell proliferation as drug screening targets, the development of anti-tumor drugs with good therapeutic effect and less toxic side effects has become an important direction of anti-tumor drug research today. Protein tyrosine kinase (protein tyrosine kinase) is a kind of protein that catalyzes the transfer of γ-phosphate on ATP to specific amino acid residues in proteins. It occupies a very important position in intracellular signal transduction pathways and regulates cell growth, A series of physiological processes such as differentiation and death. It has been shown that more than 50% of proto-oncogenes and their products have protein tyrosine kinase activity, and their abnormal expression will lead to the disorder of cell life cycle, and then lead to the occurrence of tumors. In addition, abnormal expression of tyrosine kinases is also closely related to tumor metastasis and chemotherapy resistance.

Epidermal growth factor receptor tyrosine kinase (EGFR) can mediate multiple signal transduction pathways, transmit extracellular signals into cells, and regulate the proliferation, differentiation and apoptosis of normal cells and tumor cells. Cells play an important regulatory role (Cell, 2000, 100, 113-127). Therefore, selectively inhibiting the signal transduction pathway mediated by EGFR can achieve the purpose of treating tumors and open up a feasible way for targeted therapy of tumors. Drugs targeting EGFR, such as Gefitinib, Erlotinib and Laptinib, have been marketed for the treatment of non-small cell lung cancer and breast cancer. However, clinical experience shows that most patients with non-small cell lung cancer develop drug resistance after repeated treatment with Gefitinib or Erlotinib. Drug resistance in 50% of cases is associated with an amino acid mutation in the kinase domain of EGFR (threonine residue at position 790 is mutated to methionine, T790M) (The New England Journal of Medicine, 2005, 352, 786-792) . In order to overcome the drug resistance associated with the T790M mutation, a series of irreversible ATP competitive inhibitors (such as CI-1033, BIBW2992, HKI-272, PF00299804, etc.) have entered clinical research. Irreversible inhibitors contain a Michael receptor fragment that can form a covalent bond with a conserved amino acid residue (Cys797) in the ATP binding site of EGFR, thereby obtaining a stronger EGFR binding affinity than reversible inhibitors (Journal of Medicinal Chemistry , 2009, 52, 1231-1246). Nevertheless, due to the toxic effects caused by off-target effects, side effects caused by low selectivity, and the inability to achieve sufficient drug concentrations in patients, the results of clinical trials of the above irreversible inhibitors are still not ideal (Nature, 2009, 462, 1070-1074 ). Therefore, the development of new irreversible EGFR inhibitors has great clinical significance and application prospects.

Contents of the invention

The inventors have established a virtual screening platform for EGFR-specific small molecule inhibitors by means of computer-aided drug design, comprehensively considering pharmacophore and molecular docking methods, and commercial compound databases (including ACD-3D (chemical library), ACD-3D) SC, MDDR-3D (Drug Activity Data Reporting Library) and CNPD) and found a batch of candidates with potential EGFR inhibitory activity.

The structure of the obtained candidate compounds was optimized, and a series of pteridinone compounds that had not been reported in the literature were designed and synthesized, and their structures were characterized. Molecular and cellular activity tests were carried out on this series of compounds, and a batch of compounds with high EGFR inhibitory activity were obtained. Among them, compound 006 has an IC ₅₀ inhibitory activity of EGF R ^T790M/L858 kinase of 8.4 nM, and an IC 50 inhibitory activity of HCC827 (non-small cell lung cancer cells, EGFR ^del E ^746-A750 ) cell proliferation of 0.03 _μM .

The pteridinone EGFR inhibitor involved in the present invention can block the phosphorylation process of EGFR, inhibit the growth, proliferation and differentiation of tumor cells, and thus can be developed into a new antitumor drug.

The pteridinone compound of the present invention has the structure shown in general formula I:

In the formula,

A and B are benzene rings or five-membered or six-membered heterocyclic rings with various substituents;

C is selected from any group shown below:

Wherein, X is selected from O, S and Se; R ¹ is hydrogen, a halogen atom, C1-C6 alkoxy (such as methoxy, ethoxy, etc.), optionally substituted C ₁ -C ₆ alkyl (such as halogen-substituted alkyl), optionally substituted aryl (such as halogen-substituted aryl), or optionally substituted aralkyl (such as arylmethyl);

^Each R is independently selected from hydrogen, halogen, C1-C6 alkoxy, hydroxyl, optionally substituted acyloxy, amino, optionally substituted amido, optionally substituted _C1 - _C6 alkyl, CN, Sulfonic acid, aminosulfonyl, carbamoyl, carboxyl, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, Optionally substituted piperidyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl; wherein, R _a and R _b can be selected from alkyl and chain Alkenyl;

Each ^R3 is independently selected from hydrogen, hydroxy, optionally substituted acyloxy, amino, optionally substituted amido, optionally substituted _C1 - _C4 alkyl, CN, sulfonic acid, aminosulfonyl, carboxyl , optionally substituted alkoxyformyl;

m and n are 0, 1, 2, 3 or 4 each.

In one embodiment, C is a group of the following formula:

In one embodiment, R ¹ is selected from H and alkyl.

In one embodiment, both A and B are optionally substituted phenyl.

In one embodiment, R is ^{independently} selected from H, alkoxy, morpholinyl, halogen, N-alkyl-piperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridinyl, -NR _a R _b , amido and carbamoyl (NH ₂ C(O)-), wherein, R _a and R _b can be selected from alkyl and alkenyl.

In one embodiment, R is ^{independently} selected from 4-N-methylpiperazinyl, N-morpholinyl, N-piperidinyl, N-pyrrolyl, N-pyrrolidinyl, N,N-ethyl Amino, N,N-methylmethylamino and 4-pyridyl.

In one embodiment, ^R3 is independently selected from amido, acyloxy and alkoxy.

In one embodiment, ^R3 is independently selected from the following groups:

In one embodiment, ^R is selected from the following groups:

In one embodiment, m is 1 or 2.

In one embodiment, n is 1, 2, 3 or 4.

In one embodiment, in the group represented by C, the wave shape of the part containing ^R1 is connected to C, and the other part is connected to NH.

A preferred version of the present invention is that the compound has a structure shown in general formula II:

In the formula,

Y is selected from N, CH;

Z is selected from N, ^CR6 ;

^R is hydrogen, halogen, C1-C6 alkoxy, optionally substituted _C1 - _C6 alkyl, optionally substituted aryl, optionally substituted aralkyl;

R ^is independently selected from optionally substituted acyloxy, amino, optionally substituted amido, optionally substituted C ₁ -C ₄ alkyl, CN, sulfonic acid, aminosulfonyl, carboxyl and optionally substituted Alkoxyformyl; R ⁴ , R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, halogen, C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted Amylamino, optionally substituted C ₁ -C ₆ alkyl, CN, sulfonic acid, aminosulfonyl, carbamoyl, carboxyl, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted Pyridyl; Wherein, _Ra and _R can be selected from alkyl and alkenyl; and

m is an integer of 0-3.

A more preferred version of the present invention is that the compound has a structure shown in general formula III:

In the formula,

^R is hydrogen, halogen, C1-C6 alkoxy, optionally substituted _C1 - _C6 alkyl, optionally substituted aryl, optionally substituted aralkyl

R ^is independently selected from optionally substituted acyloxy, amino, optionally substituted amido, optionally substituted C ₁ -C ₄ alkyl, CN, sulfonic acid, aminosulfonyl, carboxyl and optionally substituted Alkoxyformyl;

R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, halogen, C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted amido, optionally substituted C ₁ -C ₆ alkyl, CN, sulfonic acid, aminosulfonyl, carbamoyl, carboxyl, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl , optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl; wherein, R _a and R _can be selected from alkyl and alkenyl; and

m is 0, 1, 2 or 3. In a preferred embodiment of formula III, R ¹ is selected from H and alkyl.

In a preferred embodiment of formula III, R ^and R are ^{independently} selected from H, alkoxy, morpholinyl, halogen, N-alkyl-piperazinyl, piperidinyl, pyrrolidinyl, -NR _a R _b , amido and carbamoyl (NH ₂ C(O)—), wherein, R _a and R _b can be selected from alkyl and alkenyl.

In a preferred embodiment of formula III, R ^is selected from H, alkoxy, morpholinyl, halogen, N-alkyl-piperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, - NR _a R _b , amido and carbamoyl (NH ₂ C(O)—), wherein, R _a and R _b can be selected from alkyl and alkenyl.

In a preferred embodiment of formula III, R ^is selected from H, alkoxy, morpholinyl, halogen, N-alkyl-piperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, - NR _a R _b , amido and carbamoyl (NH ₂ C(O)-), wherein, R _a and R _b can be selected from alkyl and alkenyl; R ⁶ is H.

In a preferred embodiment of formula III, R is ^selected from halogen, 4-N-methylpiperazinyl, N-morpholinyl, N-piperidinyl, N-pyrrolyl, N-pyrrolidinyl, N , N-diethylamino, N,N-dimethylmethylamino and 4-pyridyl.

In a preferred embodiment of formula III, ^R5 and ^R6 are H, and ^R7 is amido.

In a preferred embodiment of formula III, ^R3 is independently selected from amido, acyloxy and alkoxy.

In a preferred embodiment of formula III, ^R3 is independently selected from the following groups:

In a preferred embodiment of formula III, ^R is selected from the following groups:

In a preferred embodiment of formula III, m is 1.

In a preferred embodiment of formula III, m is 1 and R is at the ⁴ position of the phenyl group.

Preferred compounds of the present invention are shown as compounds 001-032.

The present invention also includes the use of the compound of the present invention in the preparation of medicines for treating diseases mediated by epidermal growth factor receptor kinase (EGFR).

In one embodiment, the disease is cancer.

In one embodiment, the cancer is selected from non-small cell lung cancer, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, liver cancer, kidney cancer, pancreatic cancer , Colon cancer, skin cancer, leukemia, lymphoma, gastric cancer, multiple myeloma and solid tumors.

The present invention also includes the use of the compound of the present invention in the preparation of drugs for inhibiting epidermal growth factor receptor kinase (EGFR).

The above-mentioned uses of the compounds of the formula III, especially the above-mentioned uses of the compounds 002-004, 006, 008-011, 013-018, 020-027, 029-032 are particularly preferred in the present invention.

The present invention also includes pharmaceutical compositions containing the compounds of the present invention, which may optionally contain pharmaceutically acceptable carriers, excipients, diluents and the like.

Detailed ways

Some terms involved in this article are further explained as follows:

Herein, "alkyl" refers to a saturated branched or linear alkyl group with a carbon chain length of 1-10 carbon atoms. Preferred alkyl groups include 2-8 carbon atoms, 1-6, 1-4 3 carbon atoms, 3-8 carbon atoms, 1-3 carbon atoms ranging from alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, heptyl, and the like.

Alkyl groups may be substituted by one or more substituents, such as halogen or haloalkyl. For example, the alkyl group may be an alkyl group substituted with 1 to 4 fluorine atoms, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group.

Herein, "alkoxy" refers to an oxy group substituted with an alkyl group. Preferred alkoxy groups are those with a length of 1 to 6 carbon atoms, more preferably those with a length of 1 to 4 carbon atoms. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, and the like.

Herein, "alkenyl" generally means a monovalent hydrocarbon group having at least one double bond, usually containing 2-8 carbon atoms, preferably containing 2-6 carbon atoms, and may be straight or branched. Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, isopropenyl, butenyl, isobutenyl, hexenyl, and the like.

As used herein, "alkynyl" generally means a monovalent hydrocarbon group having at least one triple bond, usually containing 2-8 carbon atoms, preferably containing 2-6 carbon atoms, more usually containing 2-4 carbon atoms, which may be a straight chain or branched. Examples of alkenyl groups include ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, hexynyl, and the like.

Herein, "halogen atom" or "halogen" refers to fluorine, chlorine, bromine and iodine.

"Aryl" refers to a monocyclic, bicyclic or tricyclic aromatic group containing 6 to 14 carbon atoms, including phenyl, naphthyl, phenanthryl, anthracenyl, indenyl, fenyl, tetrahydronaphthyl, dihydronaphthyl, Hydrogenated indenyl etc. Aryl can be optionally substituted by 1-5 (for example, 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C1-4 aldehyde, C1-6 alkyl, cyano, Nitro, amino, hydroxy, hydroxymethyl, halogen-substituted alkyl (e.g. trifluoromethyl), halogen-substituted alkoxy (e.g. trifluoromethoxy), carboxy, C1-4 alkoxy, ethoxy Formyl, N(CH ₃ ) and C1-4 acyl, etc., heterocyclic or heteroaryl, etc.

Herein, "aralkyl" refers to an alkyl group substituted by an aryl group, such as a C1-C6 alkyl group substituted by a phenyl group. Examples of aralkyl groups include, but are not limited to, arylmethyl, arylethyl, and the like, such as benzyl, phenethyl, and the like.

For example, aryl can be substituted by 1-3 groups selected from: halogen, -OH, C1-4 alkoxy, C1-4 alkyl, -NO ₂ , -NH ₂ , -N(CH ₃ ) ₂ , carboxyl, and ethoxyformyl, etc.

The "5-membered or 6-membered heterocycle" used herein includes, but is not limited to, heterocyclic groups containing 1-3 heteroatoms selected from O, S and N, including but not limited to furyl, thienyl, pyrrolyl, Pyrrolidinyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, pyranyl, pyridyl, pyrimidinyl, pyrazinyl, piperidinyl, morpholinyl, etc.

As used herein, "heteroaryl" means containing from 5 to 14 ring atoms and having 6, 10 or 14 electrons shared by the ring system. And the contained ring atoms are carbon atoms and optional 1-3 heteroatoms selected from oxygen, nitrogen and sulfur. Useful heteroaryl groups include piperazinyl, morpholinyl, piperidinyl, pyrrolidinyl, thienyl, furyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, including but not limited to 2-pyridyl, 3-pyridyl and 4-pyridyl, pyrazinyl, pyrimidyl, etc.

Heteroaryl or 5-membered or 6-membered heterocyclic ring can be optionally substituted by 1-5 (for example, 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C1-4 aldehyde, C1-6 straight or branched chain alkyl, cyano, nitro, amino, hydroxyl, hydroxymethyl, halogen substituted alkyl (such as trifluoromethyl), halogen substituted alkoxy (such as trifluoromethoxy group), carboxyl, C1-4 alkoxy, ethoxyformyl, N(CH ₃ ) and C1-4 acyl.

Herein, "acyloxy" refers to a group with the structural formula "-O-C(O)-R", wherein R can be selected from alkyl, alkenyl and alkynyl. Said R can be optionally substituted.

其中，R_a和R_b可选自烷基和链烯基。Herein, "amido" refers to a group with the structural formula "-R'-NH-C(O)-R", wherein R' can be selected from a bond or an alkyl group, and R can be selected from an alkyl group or an alkenyl group , alkynyl, alkyl substituted by NR _a R _b , alkenyl substituted by NR _a R _b and alkynyl substituted by NR _a R _b , alkyl substituted by halogen, alkenyl substituted by cyano,

Wherein, R _a and R _b can be selected from alkyl and alkenyl.

Herein, "optionally substituted" means that the substituent it modifies may be optionally substituted with 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C1 -4 aldehyde, C1-6 straight chain or branched alkyl, cyano, nitro, amino, hydroxyl, hydroxymethyl, halogen substituted alkyl (such as trifluoromethyl), halogen substituted alkoxy ( Examples include trifluoromethoxy), carboxy, C1-4 alkoxy, ethoxyformyl, N( _CH3 ) and C1-4 acyl.

The present invention includes a pharmaceutical composition, which contains a therapeutically effective amount of the compound of formula I, II or III of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

Examples of pharmaceutically acceptable salts of the compounds of this invention include, but are not limited to, inorganic and organic acid salts such as hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate , fumarate, mandelate, and oxalate; and inorganic and Organic base salt.

Although the needs of each individual vary, those skilled in the art can determine the optimum dosage of each active ingredient in the pharmaceutical compositions of the present invention. Generally, the compound of the present invention or a pharmaceutically acceptable salt thereof is orally administered to mammals daily in an amount of about 0.0025 to 50 mg/kg body weight. Preferably, however, about 0.01 to 10 mg/kg is administered orally. For example, a unit oral dose may contain from about 0.01 to 50 mg, preferably from about 0.1 to 10 mg, of a compound of the invention. The unit dose may be administered one or more times per day as one or more tablets, each containing about 0.1 to 50 mg, conveniently about 0.25 to 10 mg of a compound of the present invention or a solvate thereof.

The pharmaceutical composition of the present invention can be formulated into formulations suitable for various routes of administration, including but not limited to being formulated for parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, intrathecal, intracranial Internal, nasal or external route of administration for the treatment of tumors and other diseases. The administered amount is an amount effective to ameliorate or eliminate one or more conditions. For the treatment of a particular disease, an effective amount is that amount sufficient to ameliorate or in some way alleviate symptoms associated with the disease. Such amounts may be administered as a single dose, or may be administered according to an effective treatment regimen. The amount administered may be curative, but usually the administration is to ameliorate the symptoms of the disease. Repeated dosing will generally be required to achieve the desired amelioration of symptoms. The dosage of the medicine will be determined according to the patient's age, health and weight, the type of concurrent treatment, the frequency of treatment, and the desired therapeutic benefit.

The pharmaceutical preparation of the present invention can be administered to any mammals as long as they can obtain the therapeutic effect of the compound of the present invention. The most important of these mammals is the human being.

The compound of the present invention or its pharmaceutical composition can be used to treat various diseases mediated by epidermal growth factor receptor kinase (EGFR). Herein, diseases mediated by EGFR are various cancers. The cancers include but not limited to non-small cell lung cancer, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, liver cancer, kidney cancer, pancreatic cancer, colon cancer, Skin cancer, leukemia, lymphoma, gastric cancer, multiple myeloma and solid tumors.

It should be understood that in the present invention, EGFR includes its wild type and various variants that can cause diseases. These variants include, but are not limited to, variants comprising the following mutations: T790M, L858R, L861Q, or L858R/T790M. Disease-causing truncated forms of EGFR are also encompassed by the invention. Therefore, the compound of the present invention or its pharmaceutical composition can be used to treat various diseases mediated by EGFR wild type or various variants thereof that can cause diseases, including various cancers as described above, or inhibit EGFR wild type Or the biological activity of various variants that can cause disease.

The pharmaceutical preparations according to the invention can be manufactured in a known manner. For example, manufactured by conventional blending, granulating, tableting, dissolving, or freeze-drying processes. In the manufacture of oral dosage forms, solid excipients can be combined with the active compound and the mixture optionally milled. After adding appropriate amounts of auxiliaries, if desired or necessary, the mixture of granules is processed to obtain tablets or dragee cores.

Suitable excipients are especially fillers, such as sugars such as lactose or sucrose, mannitol or sorbitol; cellulose preparations or calcium phosphates, such as tricalcium phosphate or calcium hydrogen phosphate; and binders, such as starch pastes, including corn starch , wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone. If necessary, a disintegrant can be added, such as the starch mentioned above, as well as carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, or alginic acid or its salts, such as sodium alginate. Adjuvants especially flow regulators and Lubricants such as silica, talc, stearates such as magnesium calcium stearate, stearic acid or polyethylene glycol. Dragee cores may, if desired, be provided with a suitable coating resistant to gastric juices. For this purpose, concentrated sugar solutions can be used. This solution may contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. To produce coatings resistant to gastric juices, suitable cellulose solutions, for example cellulose acetate phthalate or hydroxypropylmethylcellulose phthalate, can be used. Dyestuffs or pigments may be added to the coating of the tablets or dragee cores. For example, for identification or to characterize combinations of active ingredient doses.

Therefore, the present invention also provides a method for treating EGFR-mediated diseases or inhibiting EGFR activity, the method comprising administering the compound or pharmaceutical composition of the present invention to a subject in need.

The administration methods include but are not limited to various administration methods known in the art, which can be determined according to the actual situation of the patient. These methods include, but are not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, nasal or topical routes of administration.

The present invention also includes the use of the compounds of the present invention in the preparation of medicaments for treating EGFR-mediated diseases or inhibiting EGFR activity.

EGFR inhibitor synthesis part

The invention is further illustrated in the following examples. These examples serve only to illustrate the invention, but do not limit the invention in any way.

Reagents and conditions: (a) ArNH ₂ , DIPEA, 1,4-dioxane, rt; (b) ArNH ₂ , DIPEA, 1,4-dioxane, rt; (c) Pd/C, H ₂ , EtOH; (d )R ² COCOOEt,HOAc,EtOH,reflux;(e)trifluoroacetic acid,CH ₂ Cl ₂ ,0°C to rt;(f)acyl chloride,Et ₃ N,CH ₂ Cl ₂ ,0°C to rtor acyl chloride ,1-Methyl-2-pyrrolidinone,CH ₃ CN,0°C to rt

In the above preparation process, the definitions of R ¹ -R ⁴ are as described above. Those skilled in the art can prepare the compounds of the present invention by using various starting compounds conventionally obtained in the art as raw materials according to actual preparation needs.

Example 1

The concrete synthetic method of above-mentioned steps a-f is as follows:

Synthesis of tert-butyl (4-(2-chloro-5-nitropyrimidine-4-amino)phenyl)carbamate (step a)

Weigh 2,4-dichloro-5-nitropyrimidine (95mg, 0.49mmol) into a 10mL round bottom flask, add 3mL 1,4-oxane, stir at room temperature, and take another (4-aminophenyl ) tert-butyl carbamate (100mg, 0.48mmol) and N,N-diisopropylethylamine (69mg, 0.53mmol) were dissolved in 2mL of 1,4-dioxane and added dropwise to the above reaction solution, After the dropwise addition was completed, the mixture was stirred at room temperature for 0.5 hour, followed by TLC until the starting material was completely converted. The solvent was removed by rotary evaporation, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=10:1, v/v) to obtain (4-(2-chloro-5-nitropyrimidine-4-amino)phenyl ) tert-butyl carbamate orange solid 144mg, yield 82%. ¹ H NMR(400MHz,DMSO-d ₆ ):δ10.38(s,1H),9.46(s,1H),9.12(s,1H),7.49(d,J=8.6Hz,2H),7.39(d ,J=8.6Hz,2H),1.49(s,9H).

Synthesis of tert-butyl (4-(2-(4-methoxyphenylamino)-5-nitropyrimidine-4-amino)phenyl)carbamate (step b)

Weigh (4-(2-chloro-5-nitropyrimidine-4-amino)phenyl) tert-butyl carbamate (50mg, 0.14mmol), p-methoxyaniline (17mg, 0.14mmol), N,N - Isopropylethylamine (18mg, 0.18mmol) was placed in a 10mL round bottom flask, 5mL of 1,4-dioxane was added, stirred at room temperature for 4 hours, followed by TLC until complete conversion of the raw material. The solvent was removed by rotary evaporation, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=4:1, v/v) to obtain (4-(2-(4-methoxyphenylamino)-5-nitrate 51 mg of yellow solid tert-butyl pyrimidin-4-amino)phenyl)carbamate, yield 82%. ¹ H NMR(400MHz,DMSO-d ₆ ):δ10.30(s,1H),10.26(s,1H),9.45(s,1H),9.04(s,1H),7.49(d,J=8.8Hz ,2H),7.45(d,J=8.8Hz,2H),7.40(d,J=8.6Hz,2H),6.75(d,J=8.6Hz,2H),3.73(s,3H),1.50(s ,9H).

Synthesis of tert-butyl (4-(5-amino-2-(4-methoxyphenylamino)pyrimidine-4-amino)phenyl)carbamate (step c)

Weigh (4-(2-(4-methoxyphenylamino)-5-nitropyrimidine-4-amino)phenyl)carbamate tert-butyl ester (45mg, 0.10mmol) into a 50mL round bottom flask , add 20mL of ethanol, 5mg of palladium carbon (10%Pd), flow in hydrogen, and stir overnight at room temperature. After the reaction, suction filtration, the filtrate was spin-dried, and the crude product was purified by silica gel column chromatography (dichloromethane/methanol=5:1, v/v) to obtain (4-(5-amino-2-(4-methyl Oxyphenylamino)pyrimidine-4-amino)phenyl)carbamate tert-butyl carbamate 30mg pale pink solid, yield 83%. ¹ H NMR(400MHz,DMSO-d ₆ ):δ9.23(s,1H),8.42(s,1H),8.10(s,1H),7.62(d,J=9.2Hz,2H),7.56(s ,1H),7.53(d,J=9.2Hz,2H),7.40(d,J=8.8Hz,2H),6.77(d,J=8.8Hz,2H),3.70(s,3H),1.48(s ,9H).

Synthesis of tert-butyl (4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)carbamate (step d)

Weigh (4-(5-amino-2-(4-methoxyphenylamino)pyrimidine-4-amino)phenyl)carbamate tert-butyl ester (30mg, 0.07mmol) into a 10mL round bottom flask, Add 0.29 mL of glacial acetic acid, 5 mL of absolute ethanol, and then add ethyl glyoxylate (50% toluene solution) (16 mg, 0.08 mmol), heat to reflux and stir overnight. After the reaction, solids were precipitated and filtered with suction, the filter cake was washed with ethanol, ammonia water and deionized water, and dried. Obtained (4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)carbamate tert-butyl yellow solid 18 mg, yield 76%. ¹ H NMR(400MHz,DMSO-d ₆ ):δ10.08(s,1H),9.64(s,1H),8.84(s,1H),8.03(s,1H),7.65(d,J=8.4Hz ,2H), 7.30-7.28(m,4H), 6.61(br,2H), 3.67(s,3H), 1.52(s,9H).

Synthesis of 8-(4-aminophenyl)-2-(4-methoxyphenyl)-7(8H)-pteridinone (compound 001) (step e)

Weigh (4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)carbamate tert-butyl ester (18mg, 0.04mmol) into 5mL In a round bottom flask, add 2 mL of dichloromethane, stir at 0°C, and add 0.5 mL of trifluoroacetic acid. Stirring was then continued for 1 hour at 0°C and 1 hour at room temperature. After the reaction, add saturated sodium bicarbonate solution to neutralize until the solution is slightly alkaline, extract with dichloromethane (3×50mL), wash the organic phase with deionized water and saturated sodium chloride solution, dry over anhydrous sodium sulfate, and Solvent spin dry. 14 mg of 8-(4-aminophenyl)-2-(4-methoxyphenyl)-7(8H)-pteridinone as a yellow solid was obtained with a yield of 99%. ¹ H NMR (400MHz,DMSO-d ₆ ):δ10.04(br,1H),8.81(s,1H),8.00(s,1H),7.40(d,J=7.6Hz,2H),6.98(d , J=8.4Hz, 2H), 6.73(d, J=8.4Hz, 2H), 6.67(b r, 2H), 5.44(s, 2H), 3.70(s, 3H). ¹³ C NMR (100MHz, DMSO- _{d 6} ) _{: δ159.19,158.53,157.17,154.95,151.76,149.66,146.68,133.17,129.22,122.66,121.04,120.70,114.37,113.87,55.55 .} +H] ⁺ 361.1413, experimental value 361.1414.

Synthesis of N-(4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (compound 002) (step f)

Weigh 8-(4-aminophenyl)-2-(4-methoxyphenyl)-7(8H)-pteridinone (100mg, 0.28mmol) into a 100mL round bottom flask, add 50mL of dichloro Methane, triethylamine (28mg, 0.28mmol), stirred at 0°C, another acryloyl chloride (29mg, 0.31mmol) was dissolved in 5mL of dichloromethane, and added dropwise to the above reaction solution, after the dropwise addition was completed, under room temperature Stir overnight. The solvent was removed by rotary evaporation, and the crude product was purified by silica gel column chromatography (dichloromethane/ethyl acetate=5:1, v/v) to obtain N-(4-(2-(4-methoxyphenylamino)- 7-oxo-8(7H)-pteridinyl)phenyl)acrylamide yellow solid 34mg, yield 30%. ¹ H NMR(400MHz,DMSO-d ₆ ):δ10.42(s,1H),10.07(br,1H),8.84(s,1H),8.04(s,1H),7.87(d,J=8.8Hz ,2H),7.38(d,J=8.8Hz,2H),7.30(br,2H),6.59(br,2H),6.52(dd,J=17.0,10.0Hz,1H),6.33(dd,J= 17.0,1.8Hz,1H),5.82(dd,J=10.0,1.8Hz,1H),3.62(s,3H). ¹³ C NMR(100MHz,DMSO-d ₆ ):δ163.90,159.28,158.51,156.68,155.02 , 151.44, 146.65, 139.72, 133.00, 130.18, 129.50, 127.72, 121.02, 120.61, 113.77, 55.40. HRMS (ESI) calculated for C ₂₂ H ₁₉ N ₆ O ₃ [M+H] ⁺ 415.1519, found 415.1515.

The following compounds are all synthesized according to the method of the above-mentioned steps a-f:

N-(4-(2-(4-morpholinophenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (compound 003)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.44(s,1H),10.00(s,1H),8.82(s,1H),8.02(s,1H),7.88(d,J=8.0Hz ,1H),7.36(d,J=8.4Hz,1H),7.22(br,2H),6.59(br,2H),6.52(dd,J=17.2,10.2Hz,1H),6.33(d,J= 17.2Hz,1H),5.85(d,J=10.2Hz,1H),3.67(br,4H),2.92(br,4H).HRMS(ESI)C ₂₅ H ₂₄ N ₇ O ₃ [M+H ] ⁺ 470.1941, experimental value 470.1932.

N-(4-(2-(4-methoxyphenylamino)-6-methyl-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (compound 004)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.44(s,1H),9.90(br,1H),8.77(s,1H),7.87(d,J=8.8Hz,2H),7.50(d ,J=8.8Hz,2H),7.29(br,2H),6.59(br,2H),6.52(dd,J=17.0,10.0Hz,1H),6.33(dd,J=17.0,1.9Hz,1H) ,5.82(dd,J=10.0,1.9Hz,1H),3.61(s,3H),2.42(s,3H).HRMS(ESI)C ₂₃ H ₂₁ N ₆ O ₃ [M+H] ⁺ 429.1675 , the experimental value is 429.1671.

8-(3-aminophenyl)-2-(4-methoxyphenyl)-7(8H)-pteridinone (compound 005)

¹ H NMR (400MHz,DMSO-d ₆ ):δ10.06(br,1H),8.83(s,1H),8.01(s,1H),7.41(d,J=8.0Hz,2H),7.22(t ,J=8.0Hz,1H),6.75(d,J=7.6Hz,1H),6.67(br,2H),6.53(s,1H),6.48(d,J=7.6Hz,1H),5.35(s , 2H), 3.69 (s, 3H). HRMS (ESI) calcd for C ₁₉ H ₁₇ N ₆ O ₂ [M+H] ⁺ 361.1413, found 361.1413.

N-(3-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (Compound 006)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.42(s,1H),10.10(br,1H),8.85(s,1H),8.05(s,1H),7.84(d,J=8.0Hz ,1H),7.78(s,1H),7.56(t,J=8.0Hz,1H),7.31(br,2H),7.13(d,J=8.0Hz,1H),6.58(br,2H),6.45 (dd,J=16.8,10.4Hz,1H),6.26(dd,J=16.8,1.6Hz,1H),5.77(dd,J=10.4,1.6Hz,1H),3.65(s,3H).HRMS( _ESI ) calcd for _C22H19N6O3 [M+H] ⁺ _415.1519 , found _415.1516 .

N-(3-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)propanamide (compound 007)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.13(s,1H),10.09(s,1H),8.85(s,1H),8.04(s,1H),7.74(d,J=8.0Hz ,1H),7.71(s,1H),7.53(t,J=8.0Hz,1H),7.31(br,2H),7.07(d,J=8.0Hz,1H),6.59(br,2H),3.67 (s,3H),2.33(q,J=7.6Hz,2H),1.07(t,J=7.6Hz,3H).HRMS(ESI)C ₂₂ H ₂₁ N ₆ O ₃ [M+H] ⁺ 417.1675, the experimental value is 417.1678.

N-(4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)propanamide (Compound 008)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.15(s,1H),10.08(br,1H),8.85(s,1H),8.04(s,1H),7.80(d,J=8.4Hz ,2H),7.35-7.33(m,4H),6.61(br,2H),3.67(s,3H),2.41(q,J=7.6Hz,2H),1.14(t,J=7.6Hz,3H) . HRMS ( _ESI ) calcd. for _C22H21N6O3 [M+H] ⁺ _417.1675 , found _417.1674 .

4-(Dimethylamino)-N-(4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)-2-butene Amide (Compound 009)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.45(s,1H),10.10(br,1H),8.85(s,1H),8.05(s,1H),7.87(d,J=8.8Hz ,2H),7.37(d,J=8.8Hz,2H),7.30(br,2H),6.82(td,J=15.4,6.0Hz,1H),6.60(br,2H),6.40(d,J= 15.4Hz, 1H), 3.63(s, 3H), 3.27(d, J=5.2Hz, 2H), 2.33(s, 6H).HRMS(ESI) calculated value C ₂₅ H ₂₆ N ₇ O ₃ [M+H ] ⁺ 472.2097, experimental value 472.2095.

4-(Dimethylamino)-N-(3-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)-2-butene Amide (Compound 010)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.33(s,1H),10.08(br,1H),8.86(s,1H),8.05(s,1H),7.83(d,J=8.0Hz ,1H),7.78(s,1H),7.55(t,J=8.0Hz,1H),7.32(br,2H),7.11(d,J=8.0Hz,1H),6.74(td,J=15.2, 5.6Hz,1H),6.59(br,2H),6.30(d,J=15.2Hz,1H),3.66(s,3H),3.06(d,J=5.6Hz,2H),2.17(s,6H). HRMS ( _ESI ) calcd. for _C25H24N7O3 [M+H] ⁺ _472.2097 , found _472.2094 .

4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl acrylate (compound 011)

¹ H NMR (400MHz,DMSO-d ₆ ):δ10.15(s,1H),8.87(s,1H),8.06(s,1H),7.51(d,J=8.8Hz,2H),7.45(d ,J=8.8Hz,2H),7.31(br,2H),6.69(br,2H),6.60(dd,J=17.2,1.6Hz,1H),6.51(dd,J=17.2,9.9Hz,1H) , 6.22 (dd, J=9.9, 1.6 Hz, 1H), 3.67 (s, 3H). HRMS (ESI) calculated for C ₂₂ H ₁₈ N ₅ O ₄ [M+H] ⁺ 416.1359, found 416.1359.

4-(Dimethylamino)-N-(4-(7-oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)-2-butenamide (Compound 012)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.37(s,1H),10.19(br,1H),8.90(s,1H),8.08(s,1H),7.87(d,J=8.4Hz ,2H),7.42(d,J=7.6Hz,2H),7.38(d,J=8.4Hz,2H),7.03(br,1H),6.88(t,J=7.6Hz,1H),6.82(td ,J=15.4,5.6Hz,1H),6.37(d,J=15.4Hz,1H),3.14(d,J=5.6Hz,2H),2.24(s,6H).HRMS(ESI) calculated value C ₂₄ H ₂₄ N ₇ O ₂ [M+H] ⁺ 442.1991, found 442.1989.

4-(Dimethylamino)-N-(3-(7-oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)-2-butenamide (compound 013)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.32(s,1H),10.17(s,1H),8.90(s,1H),8.08(s,1H),7.81-7.79(m,2H) ,7.55(t,J=8.0Hz,1H),7.41(d,J=7.2Hz,2H),7.12(d,J=8.0Hz,1H),7.01(br,2H),6.87(t,J= 7.2Hz,1H),6.73(td,J=15.2,5.6Hz,1H),6.28(d,J=15.2Hz,1H),3.05(d,J=5.6Hz,2H),2.16(s,6H) . HRMS (ESI) calcd. for C ₂₄ H ₂₄ N ₇ O ₂ [M+H] ⁺ 442.1991, found 442.1996.

N-(4-(7-oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)acrylamide (Compound 014)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.44(s,1H),10.19(br,1H),8.90(s,1H),8.09(s,1H),7.88(d,J=8.4Hz ,2H),7.41-7.38(m,4H),7.03(br,2H),6.88(t,J=7.2Hz,1H),6.53(dd,J=16.8,10.4Hz,1H),6.35(dd, J=16.8, 1.6Hz, 1H), 5.84 (dd, J=10.4, 1.6Hz, 1H). HRMS (ESI) calculated for C ₂₁ H ₁₇ N ₆ O ₂ [M+H] ⁺ 385.1413, found 385.1405.

N-(3-(7-oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)acrylamide (Compound 015)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.42(s,1H),10.19(s,1H),8.91(s,1H),8.09(s,1H),7.84-7.81(m,2H) ,7.57(t,J=8.0Hz,1H),7.41(br,2H),7.15(d,J=7.6Hz,1H),7.02(br,2H),6.87(t,J=7.6Hz,1H) ,6.45(dd,J=16.8,10.4Hz,1H),6.26(dd,J=16.8,1.6Hz,1H),5.77(dd,J=10.4,1.6Hz,1H).HRMS(ESI)C ₂₁ H ₁₇ N ₆ O ₂ [M+H] ⁺ 385.1413, found 385.1413.

N-(4-(2-(4-chlorophenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (Compound 016)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.46(s,1H),10.34(s,1H),8.92(s,1H),8.11(s,1H),7.88(d,J=8.8Hz ,2H),7.41-7.36(m,4H),7.06(br,2H),6.53(dd,J=16.8,10.4Hz,1H),6.36(dd,J=16.8,1.6Hz,1H),5.84( dd, J=10.4, 1.6 Hz, 1H). HRMS (ESI) calcd for C ₂₁ H ₁₆ N ₆ O ₂ Cl [M+H] ⁺ 419.1023, found 419.1031.

N-(3-(2-(4-chlorophenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (Compound 017)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.44(s,1H),10.34(br,1H),8.93(s,1H),8.11(s,1H),7.84(s,1H),7.81 (d,J=8.4Hz,1H),7.59(t,J=8.0Hz,1H),7.43(d,J=7.2Hz,2H),7.15(d,J=7.6Hz,1H),6.46(dd ,J=16.8,10.4Hz,1H),6.26(dd,J=16.8,1.8Hz,1H),5.77(dd,J=10.12,1.8Hz,1H).HRMS(ESI)C ₂₁ H ₁₆ N ₆ O ₂ Cl [M+H] ⁺ 419.1023, experimental value 419.1027.

N-(3-(2-(4-morpholinophenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (Compound 018)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.43(s,1H),10.06(s,1H),8.84(s,1H),8.03(s,1H),7.92(br,1H),7.72 (s,1H),7.56(t,J=7.6Hz,1H),7.27(br,2H),7.12(d,J=7.2Hz,1H),6.58(br,2H),6.45(dd,J= 16.8,10.4Hz,1H),6.26(d,J=16.8Hz,1H),5.78(d,J=10.4Hz,1H),3.71(br,4H),2.94(br,4H).

N-(4-(2-(4-(4-methyl-1-piperazinyl)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (compound 019 )

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.51(s,1H),10.06(s,1H),8.83(s,1H),8.03(s,1H),7.89(d,J=8.4Hz ,2H),7.37(d,J=8.4Hz,2H),7.17(d,J=6.4Hz,1H),6.56-6.49(m,3H),6.34(d,J=16.8Hz,1H),5.85 (d, J=10.8Hz, 1H), 2.94(br, 4H), 2.37(br, 4H), 2.20(s, 3H).

N-(3-(2-(4-(4-methyl-1-piperazinyl)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (compound 020 )

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.45(s,1H),10.06(s,1H),8.84(s,1H),8.04(s,1H),7.93(br,1H),7.73 (s,1H),7.56(t,J=8.0Hz,1H),7.25(br,2H),7.12(d,J=8.0Hz,1H),6.57(br,2H),6.46(dd,J= 16.8,10.4Hz,1H),6.27(dd,J=16.8,1.8Hz,1H),5.78(dd,J=10.4,1.8Hz,1H),2.98(br,4H),2.42(br,4H), 2.22(s,3H).

N-(3-(7-oxo-2-(4-(1-piperidinyl)phenylamino)-8(7H)-pteridinyl)phenyl)acrylamide (Compound 021)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.44(s,1H),10.03(s,1H),8.83(s,1H),8.02(s,1H),7.94(br,1H),7.73 (s,1H),7.55(t,J=8.0Hz,1H),7.24(br,2H),7.11(d,J=8.0Hz,1H),6.57(br,2H),6.46(dd,J= 17.0,10.2Hz,1H),6.26(dd,J=17.0,1.8Hz,1H),5.77(dd,J=10.2,1.8Hz,1H),2.95(br,4H),1.57(br,4H), 1.49 (br,2H).

N-(3-(7-oxo-2-(4-(1-pyrrolidinyl)phenylamino)-8(7H)-pteridinyl)phenyl)acrylamide (compound 022)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.40(s,1H),9.92(s,1H),8.79(s,1H),7.99(s,1H),7.90(br,1H),7.74 (br,1H),7.54(t,J=8.0Hz,1H),7.20(br,2H),7.10(d,J=8.0Hz,1H),6.46(dd,J=17.0,10.2Hz,1H) ,6.26(dd,J=17.0,1.8Hz,1H),6.20(br,2H),5.77(dd,J=10.2,1.8Hz,1H),3.10(br,4H),1.91(br,4H).

N-(3-(2-(4-(diethylamino)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (Compound 023)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.42(s,1H),9.92(s,1H),8.80(s,1H),8.00(s,1H),7.92(br,1H),7.73 (s,1H),7.53(t,J=8.0Hz,1H),7.19(br,2H),7.09(d,J=8.0Hz,1H),6.46(dd,J=17.0,10.2Hz,1H) ,6.32(br,2H),6.27(dd,J=17.0,1.8Hz,1H),5.76(dd,J=10.2,1.8Hz,1H),3.20(br,4H),1.00(t,J=6.8 Hz, 6H).

N-(3-(2-(4-(acetylamino)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (Compound 024)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.43(s,1H),10.16(br,1H),9.78(s,1H),8.87(s,1H),8.06(s,1H),7.82 (d,J=8.0Hz,1H),7.79(s,1H),7.56(t,J=8.0Hz,1H),7.32(br,2H),7.23(br,2H),7.15(d,J= 8.0Hz,1H),6.46(dd,J=17.0,10.2Hz,1H),6.25(dd,J=17.0,1.8Hz,1H),5.76(dd,J=10.2,1.8Hz,1H),1.98( s, 3H).

4-(8-(3-acrylamidophenyl)-7-oxo-7,8-dihydropteridine-2-amino)benzamide (compound 025)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.43(s,1H),10.40(s,1H),8.95(s,1H),8.13(s,1H),7.86(s,1H),7.79 (d,J=8.0Hz,1H),7.71(br,1H),7.61(t,J=8.0Hz,1H),7.55(d,J=7.6Hz,2H),7.47(br,2H),7.18 (d,J=7.6Hz,2H),6.44(dd,J=17.0,10.2Hz,1H),6.25(dd,J=17.0,1.8Hz,1H),5.76(dd,J=10.2,1.8Hz, 1H)

N-(3-(2-(4-methoxyphenylamino)-6-methyl-7-oxo-8(7H)-pteridinyl)phenyl)acrylamide (compound 026)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.42(s,1H),9.93(br,1H),8.78(s,1H),7.83(d,J=8.0Hz,1H),7.77(s ,1H),7.56(t,J=8.0Hz,1H),7.31(br,2H),7.11(d,J=8.0Hz,1H),6.58(br,2H),6.45(dd,J=17.0, 10.2Hz, 1H), 6.26(d, J=17.0Hz, 1H), 5.77(d, J=10.2Hz, 1H), 3.65(s, 3H), 2.42(s, 3H).

N-(3-(8-(4-methoxyphenyl)-7-oxo-7,8-dihydropteridine-2-amino)phenyl)acrylamide (Compound 027)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.17(s,1H),10.01(s,1H),8.89(s,1H),8.07(s,1H),7.63(br,1H),7.33 (d,J=8.8Hz,2H),7.27(d,J=8.0Hz,1H),7.23(d,J=8.0Hz,1H),7.11(d,J=8.8Hz,2H),6.89(br ,1H),6.46(dd,J=17.0,10.2Hz,1H),6.25(dd,J=17.0,1.8Hz,1H),5.74(dd,J=10.2,1.8Hz,1H),3.85(s, 3H).

2-(3-Aminophenylamino)-8-(4-methoxyphenyl)-7(8H)-pteridinone (Compound 028)

¹ H NMR(400MHz,DMSO-d ₆ ):δ9.91(s,1H),8.85(s,1H),8.04(s,1H),7.35(d,J=8.8Hz,2H),7.16(d ,J=8.8Hz,2H),6.68-6.65(m,3H),6.16(d,J=7.2Hz,1H),4.63(s,2H),3.86(s,3H).

N-(4-(8-(4-methoxyphenyl)-7-oxo-7,8-dihydropteridine-2-amino)phenyl)acrylamide (Compound 029)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.19(br,1H),10.03(s,1H),8.87(s,1H),8.05(s,1H),7.36-7.34(m,6H) ,7.16(d,J=8.4Hz,2H),6.41(dd,J=17.0,10.2Hz,1H),6.23(dd,J=17.0,1.6Hz,1H),5.72(dd,J=10.2,1.6 Hz,1H), 3.92(s,1H).

2-(4-Aminophenylamino)-8-(4-methoxyphenyl)-7(8H)-pteridinone (compound 030)

¹ H NMR (400MHz,DMSO-d ₆ ):δ9.87(s,1H),8.77(s,1H),7.97(s,1H),7.32(d,J=8.8Hz,2H),7.13(d ,J=8.8Hz,2H),7.08(br,2H),6.24(br,2H),4.84(s,2H),3.88(s,3H).

N-(4-(2-(2-methoxy)-4-(4-methoxy-1-piperazinyl)phenylamino)-7-oxo-8(7H)-pteridinyl) Phenyl)acrylamide (Compound 031)

¹ H NMR(400MHz,DMSO-d ₆ ):δ10.43(s,1H),8.80(s,1H),8.42(s,1H),8.03(s,1H),7.85(d,J=8.6Hz ,2H),7.34(d,J=8.6Hz,2H),7.25(d,J=8.8Hz,1H),6.54-6.48(m,2H),6.33(dd,J=17.0,1.6Hz,1H) ,6.02(br,1H),5.84(dd,J=10.2,1.6Hz,1H),3.76(s,3H),3.02(br,4H),2.43(br,4H),2.23(s,3H).

N-(3-(2-(2-methoxy-4-(4-methyl-1-piperazinyl)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl ) acrylamide (compound 032)

¹ H NMR (400MHz,DMSO-d ₆ ):δ10.41(s,1H),8.80(s,1H),8.44(br,1H),8.02(s,1H),7.86(br,1H),7.71 (s,1H),7.52(t,J=8.0Hz,1H),7.30(d,J=7.6Hz,1H),7.09(d,J=8.0Hz,1H),6.53(s,1H),6.46 (dd,J=17.0,10.2Hz,1H),6.26(dd,J=17.0,1.8Hz,1H),6.02(br,1H),5.78(dd,J=10.2,1.8Hz,1H),3.76( s,3H), 3.04(br,4H), 2.44(br,4H), 2.23(s,3H).

Example 2

Biological Activity Test Section

The compound provided by the invention is carried out as follows to the in vitro inhibitory effect experiment of EGFR kinase activity:

In vitro enzyme activity analysis: wild type and various mutant types (T790M, L858R, L861Q, L858R/T790M) EGFR, Z'-Lyte Kinase Assay Kit were purchased from Invitrogen. Ten concentration gradients from 5.1×10 ^-11 mol/L to 1.0×10 ^-6 mol/L were set up for all the compounds to be tested.

The concentrations of different kinases are determined by optimization experiments, and the corresponding concentrations are: EGFR (PV3872, Invitrogen) 0.287 μg/μL, EGFR-T790M (PV4803, Invitrogen) 0.174 μg/μL, EGFR-L858R (PV4128, Invitrogen) 0.054 μg/μL , EGFR-L858R/T790M (PV4879, Invitrogen) 0.055 μg/μL. Compounds were diluted three-fold from 5.1x10 ^-9 M to 1x10 ^-4 M in DMSO. 4 μL of compound was dissolved in 96 μL of water to obtain a 4x compound solution. 40 μM ATP is dissolved in 1.33x Kinase Buffer, the Kinase/Peptide Mix contains 2x Kinase, 4 μM Tyrosine 4 Peptide is ready for use. A 10 μL kinase reaction consists of 2.5 μL compound solution, 5 μL kinase/peptide mix, 2.5 μL ATP solution. 5 µL of phosphorylated peptide solution was used instead of the kinase/peptide mix as a 100% phosphorylated control. 2.5 μL of 1.33x kinase buffer instead of ATP solution was used as 100% inhibition control, and 2.5 μL of 4% DMSO instead of compound solution was used as 0% inhibition control. The solution in the plate was mixed well and incubated at room temperature for 1.5 hours. Add 5 μL DevelopmentSolution to each well and continue to incubate at room temperature for 1 hour, during which time the non-phosphorylated peptides are cleaved. Finally, 5 μL of Stop Reagent was added to end the reaction. Pore plates were measured with an EnVision Multilabel Reader (Perkin Elmer). Experimental data were calculated using GraphPad Prism version 4.0. Each experiment was repeated more than 3 times.

Cell proliferation and growth inhibition analysis: H1975 (non-small cell lung cancer cells, EGFR ^L858R/T790M ), HCC827 (non-small cell lung cancer cells, EGFR ^{del E746-A750} ), A549 (non-small cell lung cancer cells, EGFR wild type), BT474 (Breast cancer cells, overexpressing Her2), SK-BR-3 (breast cancer cells, overexpressing Her2), MCF-7 (breast cancer cells, overexpressing Her2) cells were obtained from ATCC. Cell proliferation activity was assessed by MTS assay. Cells were exposed to the treatment conditions for 72 hours, and the number of cells used in each experiment was adjusted for each cell line according to the absorbance value (1.3-2.2 at 490 nm). Six concentration gradients (0.1nM-10μM) were set up for the compounds to be tested, and at least six groups of parallel controls were used for each concentration value.

H1975, HCC827, A549, BT474, MCF-7, SK-BR-3 cells were cultured in the corresponding medium, and the cells were passaged at least twice after recovery, and then used in experiments. Cells in log phase were trypsinized and resuspended in medium. H1975 (1000 cells per well), BT474 (1500 cells per well), MCF-7 (1500 cells per well), HCC827 (2000 cells per well), SK-BR-3 (2000 cells per well), A549 (1000 cells per well), 2000 cells) were seeded in a 96-well plate with a volume of 100 μL; 6 groups of parallel and 7 columns were set up. The plate was placed in an incubator with 5% carbon dioxide at 37°C overnight. The compounds were dissolved in DMSO to prepare a concentration of 10 μM per liter, and then the compound concentrations were gradually diluted to obtain compound concentrations of 10 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, and 0.0001 μM per liter, respectively. 2 μL of compound solution was added to 998 μL of culture medium, and the mixture was mixed well. 100 μL of the mixture was added to a 96-well plate. 2 μL of DMSO instead of the compound solution was used as a 0% inhibition control. After 68 hours of incubation, 20 μL of MTT (5 mg/mL) was added. After 4 hours, discard the supernatant and add 150 μL DMSO. After shaking for 10 minutes, the plate was read with Synergy HT (Bio TeK) (OD490). Data were calculated using GraphPad Prism version 4.0, and _IC50 values were adjusted by non-linear regression models using dose-response curves.

The test results are shown in Tables 1 and 2 below.

Table 2

Claims (10)

1. the compound that there is structure shown in general formula I:

In formula,

A and B are for being with various substituent phenyl ring or five yuan or hexa-member heterocycle;

C is selected from arbitrary group as follows:

Wherein, X is selected from O, S and Se; R ¹For hydrogen, halogen atom, C1-C6 alkoxyl group, the optional C replaced ₁-C ₆Alkyl, the optional aryl replaced or the optional aralkyl replaced;

R ²Independently be selected from separately hydrogen, halogen, C1-C6 alkoxyl group, hydroxyl, the optional acyloxy replaced, amino, the optional amido replaced, the optional C replaced ₁-C ₆The pyrrolidyl of the phenyl of alkyl, CN, sulfonic group, amino-sulfonyl, formamyl, carboxyl, the optional alkoxyl formyl replaced, optional replacement, the optional N-alkylpiperazinyl replaced, the optional morpholinyl replaced, the piperidyl optionally replaced, the optional pyrryl replaced, optional replacement ,-NR _aR _b, the optional pyridyl replaced;

R ³Independently be selected from separately hydrogen, hydroxyl, the optional acyloxy replaced, amino, the optional amido replaced, the optional C replaced ₁-C ₄Alkyl, CN, sulfonic group, amino-sulfonyl, carboxyl, the optional alkoxyl formyl replaced;

R _aAnd R _bIndependently be selected from separately alkyl and alkenyl; With

M and n respectively do for oneself 0,1,2,3 or 4.

2. compound as claimed in claim 1, is characterized in that, described compound has structure shown in general formula I I:

In formula,

Y is selected from N, CH;

Z is selected from N, CR ⁶

R ¹For hydrogen, halogen, C1-C6 alkoxyl group, the optional C replaced ₁-C ₆Alkyl, the optional aryl replaced, the optional aralkyl replaced;

R ³Independently be selected from acyloxy, amino, the optional amido replaced, the optional C replaced of optional replacement ₁-C ₄Alkyl, CN, sulfonic group, amino-sulfonyl, carboxyl and the optional alkoxyl formyl replaced;

R ⁴, R ⁵, R ⁶And R ⁶Independently be selected from separately hydrogen, halogen, C ₁-C ₆Alkoxyl group,, hydroxyl, the optional acyloxy replaced, amino, the optional amido replaced, the optional C replaced ₁-C ₆The pyrrolidyl of the phenyl of alkyl, CN, sulfonic group, amino-sulfonyl, formamyl, carboxyl, the optional alkoxyl formyl replaced, optional replacement, the optional N-alkylpiperazinyl replaced, the optional morpholinyl replaced, the piperidyl optionally replaced, the optional pyrryl replaced, optional replacement ,-NR _aR _b, the optional pyridyl replaced;

R _aAnd R _bIndependently be selected from separately alkyl and alkenyl; With

M and n respectively do for oneself 0,1,2,3 or 4.

3. compound as claimed in claim 1, is characterized in that, described compound has structure shown in general formula III:

In formula,

R ¹For hydrogen, halogen, C1-C6 alkoxyl group, the optional C replaced ₁-C ₆Alkyl, the optional aryl replaced, the optional aralkyl replaced;

R ³Independently be selected from acyloxy, amino, the optional amido replaced, the optional C replaced of optional replacement ₁-C ₄Alkyl, CN, sulfonic group, amino-sulfonyl, carboxyl and the optional alkoxyl formyl replaced;

R ⁵, R ⁶And R ⁷Independently be selected from separately hydrogen, halogen, C ₁-C ₆Alkoxyl group,, hydroxyl, the optional acyloxy replaced, amino, the optional amido replaced, the optional C replaced ₁-C ₆The pyrrolidyl of the phenyl of alkyl, CN, sulfonic group, amino-sulfonyl, formamyl, carboxyl, the optional alkoxyl formyl replaced, optional replacement, the optional N-alkylpiperazinyl replaced, the optional morpholinyl replaced, the piperidyl optionally replaced, the optional pyrryl replaced, optional replacement ,-NR _aR _b, the optional pyridyl replaced;

R _aAnd R _bIndependently be selected from separately alkyl and alkenyl; With

M and n respectively do for oneself 0,1,2,3 or 4.

In a preferred embodiment of formula III, R ¹Be selected from H and alkyl.

4. compound as claimed in claim 3, is characterized in that,

R ¹Be selected from H and C1-C6 alkyl;

R ³Be selected from amido, acyloxy and alkoxyl group;

R ⁵Be selected from H, alkoxyl group, morpholinyl, halogen, N-alkyl-piperazinyl, piperidyl, pyrryl, pyrrolidyl, pyridyl ,-NR _aR _b, amido and formamyl, wherein, R _aAnd R _bCan be selected from alkyl and alkenyl;

R ⁶Be selected from H; With

R ⁷Be selected from H or amido.

5. compound as claimed in claim 4, is characterized in that,

R ⁵Be selected from halogen, 4-N-methylpiperazine base, N-morpholinyl, N-piperidyl, N-pyrryl, N-pyrrolidyl, N, N-ethylamino, N, N-methyl methylamino and 4-pyridyl;

R ³Be selected from any in following group:

R ⁷For H.

6. be selected from following compound:

7. a pharmaceutical composition, is characterized in that, described pharmaceutical composition contains the described compound of any one or its pharmacy acceptable salt in claim 1-6, and pharmaceutically acceptable carrier or vehicle.

8. the purposes in the disease that in claim 1-6, the described compound of any one is mediated by epidermal growth factor receptor kinase in the preparation treatment or the medicine that suppresses epidermal growth factor receptor kinase.

9. purposes as claimed in claim 8, is characterized in that, described disease is cancer.

10. purposes as claimed in claim 9, is characterized in that, described cancer is selected from nonsmall-cell lung cancer, mammary cancer, prostate cancer, neurogliocytoma, ovarian cancer, incidence squama cancer, cervical cancer, the esophageal carcinoma, liver cancer, kidney, carcinoma of the pancreas, colorectal carcinoma, skin carcinoma, leukemia, lymphoma, cancer of the stomach, multiple marrow cancer and solid tumor.