CN105985342A - Pyrimido pyrimidine dione derivative as EGFR inhibitor and application thereof - Google Patents

Pyrimido pyrimidine dione derivative as EGFR inhibitor and application thereof Download PDF

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CN105985342A
CN105985342A CN201510064374.4A CN201510064374A CN105985342A CN 105985342 A CN105985342 A CN 105985342A CN 201510064374 A CN201510064374 A CN 201510064374A CN 105985342 A CN105985342 A CN 105985342A
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compound
cancer
alkyl
egfr
substituted
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CN105985342B (en
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李洪林
徐玉芳
谢华
赵振江
郝永佳
王霞
丁健
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East China University of Science and Technology
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East China University of Science and Technology
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Priority to CN201680008995.3A priority patent/CN107428763B/en
Priority to PCT/CN2016/073726 priority patent/WO2016124160A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/541—Non-condensed thiazines containing further heterocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Abstract

The invention relates to a pyrimido pyrimidine dione derivative as an EGFR inhibitor and an application thereof. Concretely, the invention relates to a compound shown as a formula I, a pharmaceutical composition containing the compound shown as the formula I, and a purpose of the compound in preparation of a medicine for treating EGFR-related diseases or inhibiting EGFR.

Description

Pyrimido-pyrimidine derovatives and application thereof as EGFR inhibitor
Technical field
The present invention relates to medicinal chemistry art；Specifically, the present invention relates to novel pyrimido-pyrimidine diketone Derivant, its synthetic method and as EGFR inhibitor in the medicine preparing tumor-related illness should With.
Background technology
Cancer is also known as malignant tumor, is the big class disease as feature with abnormal cell proliferation and transfer, has The feature that sickness rate is high and mortality rate is high, is to threaten human health, causes one of malignant disease of death.Grind Studying carefully data to show, there are 12,700,000 cancer patients in the whole world in 2008, and wherein death toll is up to more than 700 ten thousand.And The new tumour patient of sending out in the whole world 20% is in China, and the tumor mortality patient of 24% is in China.If do not taked Effective measures are prevented, or take out more excellent therapeutic scheme, it is contemplated that to the year two thousand thirty, will go out every year in world wide Existing 26,000,000 increase cases of cancer newly, and number of cancer deaths is up to 17,000,000.In existing cancer, pulmonary carcinoma It is the malignant tumor that M & M is the highest in current world wide, wherein nonsmall-cell lung cancer (NSCLC) more than the 80% of patients with lung cancer is accounted for.Predict according to World Health Organization (WHO) (WHO), by 2025, I State increases cases of lung cancer every year newly will be more than 1,000,000.Once being diagnosed as pulmonary carcinoma, patient just only has remote life Depositing prospect, within 5 years, survival rate is less than 15%.
From the eighties in 20th century, along with going deep into of oncomolecularbiology research, tumorigenesis Molecular mechanism the most clear.In the factor of many induction cancers, cancerous cell is caused by gene mutation Some protein kinase of high expressed is one of principal element causing its signal transduction pathway exception.Protein cheese Histidine kinase is the important factor in signal transduction process, participates in a series of cellular activity, grow with cell, Differentiation, breed closely related.The tyrosine that the γ phosphate of its catalysis ATP transfers to many key proteins is residual On base, make phenolic hydroxyl group phosphorylation, thus transmit signal.Therefore, selective kinases inhibitor is developed Block or regulate and control and open owing to the abnormal disease produced of these signal paths has been considered as antitumor drug The effective research strategy sent out.In numerous tyrosine kinase, epidermal growth factor recipient tyrosine Kinases (epidermal growth factor receptor tyrosine kinase, EGFR) is indispensable important Ingredient.EGFR is made up of 1186 aminoacid, and encoding a molecular weight is the transmembrane glycopeptide of 170-kDa In vain.EGFR can mediate a plurality of signal transduction pathway, and extracellular signal is delivered to intracellular, to normal cell With the propagation of tumor cell, differentiation and apoptosis all play important regulation effect (Cell, 2000,100, 113-127).EGFR is constructive expression's composition of many normal epithelial tissues (such as skin and hair follicle), and In major part solid tumor, there is process LAN or high expressed in EGFR.Such as, in pulmonary carcinoma, the table of EGFR The rate of reaching reaches 40～80%.The most optionally suppress EGFR, disturb the signal transduction pathway that it mediates, can To reach to treat the purpose of pulmonary carcinoma, open a practical way for targeted therapy of lung cancer.
On clinical treatment, in conjunction with traditional radiotherapy, chemotherapy, with EGFR targeted drug such as gefitinib (Iressa), Erlotinib (Tarceva) etc. carries out first-line drug and is proved to be very effective in lung cancer therapy.But, Clinical practice shows: major part Patients with Non-small-cell Lung treats it at use gefitinib or Erlotinib After, within the 6-12 month, acquired drug-resistance can occur.The drug resistance of the most about 50% case is tied with EGFR kinases In structure territory, the sudden change (790 threonine residues sport methionine, T790M) of an amino acid residue is relevant (The New England Journal of Medicine,2005,352,786-792).T790M sudden change causes pressing down Preparation produce when EGFR is combined sterically hindered or increase EGFR Yu ATP affinity so that this kind of can The anticancer effect of the competitive inhibitor that inverse property combines weakens significantly.The generation of drug resistance not only reduces patient Sensitivity to medicine, has been greatly reduced the life quality of tumor patient.In order to overcome T790M sudden change to draw The drug resistance risen, a series of irreversible ATP competitive inhibitors are (such as CI-1033, HKI-272, PF00299804 Deng) have been enter into clinical investigation phase.Irreversible inhibitor contains a michael acceptor fragment, energy and EGFR Conservative amino acid residues (Cys797) of ATP-binding site form covalent bond, thus obtain ratio can The EGFR binding affinity that inverse property inhibitor is higher.While it is true, owing to this type of medicine to wild type and is dashed forward Modification EGFR selectivity is poor, and therefore its maximum tolerated dose (MTD) is relatively low, and clinical experiment effect is the most inconspicuous.
Therefore, research and development Selective depression T790M sudden change, overcome the third generation EGFR target of clinical drug-resistant To medicine, there is great clinical meaning and application prospect.
Summary of the invention
It is an object of the invention to provide can Selective depression T790M sudden change pyrimido-pyrimidine diketone derivatives Thing is as EGFR inhibitor.
It is a further object of the present invention to provide the pharmaceutical composition comprising above-claimed cpd.
Further object of the present invention is to provide above-claimed cpd in preparation treatment EGFR relevant disease or suppression Purposes in the medicine of EGFR.
In first aspect, the present invention provides the compound shown in formula I or its pharmaceutically acceptable salt:
In formula, A is phenyl ring, five yuan or hexa-member heterocycle, C3-C8Cycloalkyl；
R1It is each independently selected from hydrogen, halogen, C1-C3Alkoxyl, C1-C3Alkyl, C1-C4Alkylamidoalkyl, Substituted piperazinyl, substituted homopiperazine base, substituted morpholinyl, substituted thio-morpholinyl, 4-N-first Base piperazinyl, 4-N-acetylpiperazinyl, 4-N, N-lupetidine base, substituted piperidyl ,-NRaRb, Wherein, RaAnd RbAlkyl can be independently selected from and containing azanyl；
R2It is each independently selected from following group:
R3Selected from lower group: hydrogen, C1-C10Alkyl, substituted C1-C10Alkyl, C2-C6Alkenyl, C2-C6Alkynes Base, optionally substituted C3-C8Cycloalkyl, optionally substituted aryl, optionally substituted benzyl, optionally substituted Heterocyclic radical, optionally substituted aromatic heterocyclic；
B is selected from lower group:
M is the integer of 0-7.
In a particular embodiment, described compound is as shown in formula II:
In formula, B, R1、R2、R3As defined above；
M is the integer of 0-5.
In a particular embodiment, described compound is as shown in general formula III:
In formula,
R2It is selected from
R3Selected from C1-C6Alkyl, preferably methyl or isopropyl；Or the substituted C of phenyl1-C6Alkyl, preferably benzyl Base；
R4、R5、R6、R7And R8It is independently selected from lower group:
In a particular embodiment, R5And R8For H；Or, R5、R7And R8For H.
In a particular embodiment, the present invention provides selected from the compound of lower group or it is pharmaceutically acceptable Salt:
In second aspect, the present invention provides a kind of pharmaceutical composition, and described pharmaceutical composition contains the present invention the Compound described in Yi Fangmian or its pharmaceutically acceptable salt, and pharmaceutically acceptable carrier or figuration Agent.
In a preferred embodiment, described pharmaceutical composition is adapted for the dosage form being administered orally, and includes but not limited to Tablet, solution, suspension, capsule, granule, powder.
In the third aspect, the present invention provides the compound described in first aspect present invention in preparation treatment or prevention Purposes in the disease of EGFR mediation, or the medicine of suppression EGFR.
In a particular embodiment, the disease of described EGFR mediation is cancer.
In a particular embodiment, described cancer be selected from lower group: nonsmall-cell lung cancer, small cell lung cancer, Adenocarcinoma of lung, lung squamous cancer, breast carcinoma, carcinoma of prostate, neurogliocytoma, ovarian cancer, G. cephalantha, Cervical cancer, the esophageal carcinoma, hepatocarcinoma, renal carcinoma, cancer of pancreas, colon cancer, skin carcinoma, leukemia, lymphoma, Gastric cancer, multiple bone marrow cancer and solid tumor.
In fourth aspect, the present invention provides and utilizes the compounds for treating described in first aspect present invention or prevention The disease method of EGFR mediation.
In a preferred embodiment, the disease of described EGFR mediation is cancer；Preferably, described cancer Selected from lower group: nonsmall-cell lung cancer, small cell lung cancer, adenocarcinoma of lung, lung squamous cancer, breast carcinoma, carcinoma of prostate, Neurogliocytoma, ovarian cancer, G. cephalantha, cervical cancer, the esophageal carcinoma, hepatocarcinoma, renal carcinoma, pancreas Cancer, colon cancer, skin carcinoma, leukemia, lymphoma, gastric cancer, multiple bone marrow cancer and solid tumor.
In should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and below (such as embodiment) Can be combined with each other between each technical characteristic of middle specific descriptions, thus constitute new or preferred technical side Case.As space is limited, the most tired at this state.
Detailed description of the invention
Inventor is through extensively in-depth study, it was unexpectedly found that the brand-new pyrimido of a collection of structure is phonetic Pyridine derovatives, these derivants can be suddenlyd change by Selective depression EGFR T790M, right EGFRT790M/L858RThe IC of kinase inhibiting activity50Value reaches nM rank；To cancerous cell (EGFRL858R/T790M Sudden change) inhibitory activity IC bred50Value reaches a μM rank.Complete the present invention on this basis.
The present inventor has synthesized the candidate compound with EGFR inhibitory activity.To the candidate compound obtained Carry out structure optimization, design and synthesize a series of pyrimido [4,5-d] pyrimidine having no that document is reported -2,4 (1H, 3H)-cyclohexadione compounds, and carry out finishing structure sign.This series compound has been carried out molecular water The active testing of gentle cellular level, obtains a collection of chemical combination with Selective depression EGFR T790M sudden change Thing.Wherein compound 007 is to EGFRT790M/L858Kinase inhibiting activity IC50(non-little for 30.0nM, H1975 Cell lung cancer cell, EGFRL858R/T790M) cell inhibitory effect activity IC50It it is 0.23 μM.
Term defines
Some group definition arrived referred to herein are as follows:
Herein, " alkyl " refers to the saturated branched-chain or straight-chain alkyl that carbon chain lengths is 1-10 carbon atom, excellent The alkyl of choosing includes long 2-8,1-6,1-4,3-8,1-3 carbon atom alkyl not etc..Alkyl Example include but not limited to: methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group, heptyl Deng.Alkyl can be replaced by one or more substituent groups, such as, replaced by halogen or haloalkyl.Such as, The alkyl that alkyl can be replaced by 1-4 fluorine atom, or the alkane that alkyl can be replaced by fluoro-alkyl Base.
Herein, " alkoxyl " refers to the epoxide replaced by alkyl.Preferably alkoxyl is long 1-6 carbon atom Alkoxyl, the alkoxyl of the longest 1-4 carbon atom, the alkoxyl of the longest 1-3 carbon atom. The example of alkoxyl includes but not limited to: methoxyl group, ethyoxyl, propoxyl group etc..Alkoxyl can be by 1 Or multiple substituent group replaces, such as, replaced by halogen or haloalkyl.Such as, alkoxyl can be by 1-4 The substituted alkyl of individual fluorine atom, or the alkyl that alkyl can be replaced by fluoro-alkyl.
Herein, " alkenyl " generally represents the univalence hydrocarbyl with at least one double bond, usually contains 2-8 Individual carbon atom, preferably comprises 2-6 carbon atom, can be straight or branched.The example of alkenyl include but not It is limited to vinyl, acrylic, isopropenyl, cyclobutenyl, isobutenyl, hexenyl etc..
Herein, " alkynyl " generally represents the univalence hydrocarbyl with at least one three key, usually contains 2-8 carbon Atom, preferably comprises 2-6 carbon atom, more generally contains 2-4 carbon atom, can be straight or branched. The example of alkenyl includes acetenyl, propinyl, isopropynyl, butynyl, butynyl, hexin base etc..
Herein, " halogen " refers to fluorine, chlorine, bromine or iodine.
Herein, " aryl " refers to the monocycle containing 6 to 14 carbon atoms, dicyclo or tricyclic aromatic group, including Phenyl, naphthyl, phenanthryl, anthryl, indenyl, base, tetrahydro naphthyl, indanyl etc..Aryl can Optionally replaced selected from following substituent group by 1-5 (such as, 1,2,3,4 or 5): halogen, C1-4 Aldehyde radical, C1-6Alkyl, cyano group, nitro, amino, hydroxyl, methylol, halogen substiuted alkyl (such as Trifluoromethyl), the alkoxyl (such as trifluoromethoxy) of halogen substiuted, carboxyl, C1-4Alkoxyl, second Oxygen formoxyl, N (CH3) and C1-4Acyl group etc., heterocyclic radical or heteroaryl etc..
" heterocyclic radical " used herein includes but not limited to containing 1-3 selected from heteroatomic 5 yuan or 6 of O, S or N Unit's heterocyclic group, includes but not limited to furyl, thienyl, pyrrole radicals, pyrrolidinyl, pyrazolyl, miaow Oxazolyl, triazolyl, oxazolyl, pyranose, pyridine radicals, pyrimidine radicals, pyrazinyl, piperidyl, morpholinyl Deng.
" aromatic heterocyclic " used herein refers to containing 5-14 annular atoms, and has 6,10 or 14 electronics Member ring systems shares.And contained annular atoms be carbon atom and 1-3 optional from oxygen, nitrogen, sulfur miscellaneous former Son.Useful aromatic heterocyclic includes piperazinyl, morpholinyl, piperidyl, pyrrolidinyl, thienyl, furan Base, pyranose, pyrrole radicals, imidazole radicals, pyrazolyl, pyridine radicals, include, but are not limited to 2-pyridine radicals, 3-pyridine radicals and 4-pyridine radicals, pyrazinyl, pyrimidine radicals etc..
Aromatic heterocyclic is optionally selected from following substituent group by 1-5 (such as, 1,2,3,4 or 5) Replace: halogen, C1-4Aldehyde radical, C1-6Straight or branched alkyl, cyano group, nitro, amino, hydroxyl, hydroxyl first Base, the alkyl (such as trifluoromethyl) of halogen substiuted, the alkoxyl (such as trifluoromethoxy) of halogen substiuted, Carboxyl, C1-4Alkoxyl, ethoxycarbonyl, N (CH3) and C1-4Acyl group.
Herein, " acyloxy " refers to the group that structural formula is "-O-C (O)-R ", wherein, R be selected from alkyl, Alkenyl and alkynyl.Described R is optionally replaced.
Herein, " acylamino-" refers to the group that structural formula is "-R '-NH-C (O)-R ", and wherein, R ' is selected from hydrogen Or alkyl, R be selected from alkyl, alkenyl, alkynyl, by NRcRdSubstituted alkyl, by NRcRdSubstituted Alkenyl and NRcRdSubstituted alkynyl, the alkyl being optionally substituted by halogen, the alkenyl that replaced by cyano group, wherein, RcAnd RdIt is selected from alkyl and alkenyl.
Herein, " optionally substituted " refers to that its substituent group modified is optionally by 1-5 (such as, 1,2, 3,4 or 5) replace selected from following substituent group: halogen, C1-4Aldehyde radical, C1-6Straight or branched alkyl, Cyano group, nitro, amino, hydroxyl, methylol, the alkyl (such as trifluoromethyl) of halogen substiuted, halogen take The alkoxyl (such as trifluoromethoxy) in generation, carboxyl, C1-4Alkoxyl, ethoxycarbonyl, N (CH3) and C1-4 Acyl group.
The compound of the present invention
The compound of the present invention is the compound shown in below general formula I or its pharmaceutically acceptable salt:
In formula, A is phenyl ring, five yuan or hexa-member heterocycle, C3-C8Cycloalkyl；
R1It is each independently selected from hydrogen, halogen, C1-C3Alkoxyl, C1-C3Alkyl, C1-C4Alkylamidoalkyl, Substituted piperazinyl, substituted homopiperazine base, substituted morpholinyl, substituted thio-morpholinyl, 4-N-first Base piperazinyl, 4-N-acetylpiperazinyl, 4-N, N-lupetidine base, substituted piperidyl ,-NRaRb, Wherein, RaAnd RbAlkyl can be independently selected from and containing azanyl；
R2It is each independently selected from following group:
R3Selected from lower group: hydrogen, C1-C10Alkyl, substituted C1-C10Alkyl, C2-C6Alkenyl, C2-C6Alkynes Base, optionally substituted C3-C8Cycloalkyl, optionally substituted aryl, optionally substituted benzyl, optionally substituted Heterocyclic radical, optionally substituted aromatic heterocyclic；
B is selected from lower group:
M is the integer of 0-7.
In a particular embodiment, A ring is phenyl ring, thus the compound of the present invention is as shown in below general formula II:
In formula, B, R1、R2、R3As mentioned above；It is the integer of 0-5 with m.
In a preferred embodiment, the above-mentioned phenyl ring in the compound of the present invention can be to replace or unsubstituted , such as, the compound of the present invention can be as shown in below general formula III:
In formula,
R2It is selected from
R3Selected from C1-C6Alkyl, preferably methyl or isopropyl；Or the substituted C of phenyl1-C6Alkyl, preferably benzyl Base；
R4、R5、R6、R7And R8It is independently selected from lower group:
In further embodiment, the above-mentioned phenyl ring in the compound of the present invention can make ortho position replace, Meta replaces and/or para-orientation.In a preferred embodiment, the above-mentioned phenyl ring in the compound of the present invention It is that ortho position replaces and para-orientation.In another preferred embodiment, upper in the compound of the present invention Stating phenyl ring is that ortho position replaces, meta replaces and para-orientation.In a particular embodiment, formula above R in III5And R8For H；Or, R5、R7And R8For H.
The present inventor's synthesis has obtained pyrimido [4,5-d] pyrimidine that a series of structure has no that document is reported -2,4 (1H, 3H)-cyclohexadione compounds, concrete compound is as follows:
On the basis of the compound of the present invention, the present invention provides a kind of pharmaceutical composition, and said composition contains The compound of the present invention of therapeutically effective amount or its pharmaceutically acceptable salt, and pharmaceutically acceptable load Body or excipient.
The example of the pharmaceutically acceptable salt of the compounds of this invention includes but not limited to inorganic and acylate, Such as hydrochlorate, hydrobromate, sulfate, citrate, lactate, tartrate, maleate, Fumarate, mandelate and oxalates；And with alkali such as sodium hydroxyl, three (hydroxymethyl) aminomethane Inorganic and the organic alkali salt that (TRIS, amine butantriol) and N-METHYL-ALPHA-L-GLUCOSAMINE are formed.
Although each Man's Demands is different, those skilled in the art can determine that in pharmaceutical composition of the present invention The optimal dose of every kind of active component.Generally, the compound of the present invention or it is pharmaceutically acceptable Salt, to mammal oral administration every day, dose is according to about 0.0025 to 50 mg kg of body weight.It is desirable that It it is per kilogram oral administration about 0.01 to 10 milligrams.Such as, unit oral doses can include about 0.01 to 50 Milligram, the compounds of this invention of preferably about 0.1 to 10 milligrams.Unit dose can give one or many, often It is one or more pieces, and every contains about 0.1 to 50 milligrams, of the present inventionization of the most about 0.25 to 10 milligrams Compound or its solvate.
The pharmaceutical composition of the present invention can be formulated into the dosage form of applicable various route of administration, including but not It is limited to be formulated for parenteral, subcutaneous, vein, muscle, intraperitoneal, transdermal, oral cavity, in sheath, cranium In, the form that nasal cavity or topical route are administered, it is used for treating tumor and other diseases.Dosage is effectively Improve or eliminate the dose of one or more disease.For the treatment of specified disease, effective dose is to be enough to improve Or alleviate the dose of the symptom relevant with disease in some manner.Such dose can be executed as single dose With, or can be administered according to effective therapeutic scheme.Dosage also permits cure diseases, but is administered generally It it is the symptom in order to improve disease.Improve it is generally required to repetitively administered realizes required symptom.The dosage of medicine By the age according to patient, healthy and body weight, the kind of concurrent treatment, the frequency for the treatment of, and needed for control Curative effect benefit determines.
The pharmaceutical preparation of the present invention can give any mammal, as long as they can obtain the compounds of this invention Therapeutic effect.The most importantly mankind in these mammals.
The compound of the present invention or its pharmaceutical composition can be used for treating and various swashed by EGF-R ELISA The disease that enzyme (EGFR) mediates.Herein, EGFR the disease mediated is various cancer.Described cancer bag Include but be not limited to: nonsmall-cell lung cancer, small cell lung cancer, adenocarcinoma of lung, lung squamous cancer, breast carcinoma, prostate Cancer, neurogliocytoma, ovarian cancer, G. cephalantha, cervical cancer, the esophageal carcinoma, hepatocarcinoma, renal carcinoma, Cancer of pancreas, colon cancer, skin carcinoma, leukemia, lymphoma, gastric cancer, multiple bone marrow cancer and solid tumor.
The pharmaceutical preparation of the present invention can manufacture in a known manner.Such as, by traditional mixing, pelletize, system Ingot, dissolves, or freezing dry process manufacture.When manufacturing oral formulations, can be in conjunction with solid adjuvant material and activation Compound, selectivity milled mixtures.If after needing or adding appropriate amount of addition agent if desired, processing granule mixing Thing, it is thus achieved that tablet or lozenge core.
Suitably adjuvant particularly filler, such as saccharide such as lactose or sucrose, mannitol or sorbitol；Fiber Element preparation or calcium phosphate, such as tricalcium phosphate or calcium hydrogen phosphate；And binding agent, such as gelatinized corn starch, bag Include corn starch, wheaten starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, Hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, or polyvinylpyrrolidone.If it is required, can increase Disintegrating agent, ratio starch as mentioned above, and carboxymethyl starch, crospolyvinylpyrrolidone, agar, Or alginic acid or its salt, such as sodium alginate.Adjuvant particularly flowing regulator and lubricant, such as, silicon Stone, Talcum, stearates, such as magnesium calcium stearate, stearic acid or Polyethylene Glycol.It is possible if desired to Lozenge nuclear core provides the suitable coating that can resist gastric juice.To this end, concentration saccharide solution can be applied.This Individual solution can contain Radix Acaciae senegalis, Talcum, polyvinylpyrrolidone, Polyethylene Glycol and/or titanium dioxide, Paint solution and suitable organic solvent or solvent mixture.In order to prepare the coating of resistant to gastric juice, can use suitably Cellulose solution, such as cellulose acetate phthalic acid or hydroxypropyl methyl cellulose phthalic acid. Dyestuff or pigment can be added to the coating of tablet or lozenge nuclear core.Such as, for identifying or in order to characterize activity The combination of component dose.
Based on above-claimed cpd and pharmaceutical composition, the present invention further provides and a kind of treat EGFR mediation The method of disease, the method includes that the object giving needs is with the compound of the present invention or pharmaceutical composition.
Medication includes but not limited to various medications well known in the art, can be according to the actual feelings of patient Condition is determined.These methods include but not limited to parenteral, subcutaneous, vein, muscle, intraperitoneal, transdermal, Oral cavity, in sheath, intracranial, nasal cavity or topical route be administered.
The present invention also includes that the compounds of this invention in preparation prevention or treats disease or the suppression that EGFR mediates Purposes in the medicine of activity of EGFR.
Advantages of the present invention:
1. the compound that the present invention provides be brand-new pyrimido [4,5-d] pyrimidine-2,4 (1H, 3H) of a kind of structure- Cyclohexadione compounds；
2. the compound that the present invention provides has excellence to the cancerous cell that mutant egf R or EGFR suddenlys change Inhibitory activity；
3. the compound that the present invention provides is that exploitation can suddenly change by Selective depression T790M, can overcome clinical resistance to The EGFR targeted drug of medicine is laid a good foundation, and possesses great industrialization and commercialization prospect and market price Value, remarkable in economical benefits.
Below in conjunction with being embodied as case, technical scheme is further described, but following case study on implementation It is not construed as limiting the invention, the various sides of using that all principles according to the present invention and technological means use Method, belongs to the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, generally according to Normal condition, or according to the condition proposed by manufacturer.Unless otherwise indicated, otherwise percentage ratio and number Calculate by weight.
Materials and methods
The synthesis of pyrimido [4,5-d] pyrimidine-2,4 (1H, the 3H)-cyclohexadione compounds of the present invention is as follows:
Reagent and condition: (a) DIPEA, CH3CN, backflow；(b)ArNH2,CH3CN, backflow；(c)NaOH, THF, backflow；(d)R3NH2,HATU,DIPEA,DMF；(e)K2CO3, CDI, THF, backflow；(f) three Fluoroethanoic acid, CH2Cl2, 0 DEG C-room temperature；(g) acyl chlorides, Et3N,CH2Cl2, 0 DEG C of room temperature.
Embodiment 1
The concrete synthetic method of above-mentioned steps a-g is as follows:
The synthesis of 1.4-((3-((tert-butoxycarbonyl) amino) phenyl) amino)-2-chloropyrimide-5-Ethyl formate
Weigh 2,4-bis-chloro-5-nitro-pyrimidine (2.210g, 10mmol), DIPEA (1.290g, 10mmol) in 50mL single port flask, adds 15mL acetonitrile and dissolves.Separately take (3-aminophenyl) t-butyl carbamate (2.080 G, 10mmol) it is dissolved in 10mL acetonitrile, it is added drop-wise in above-mentioned reactant liquor, drips backflow 2h.TLC follows the tracks of Converting to raw material, be cooled to room temperature, sucking filtration, acetonitrile washs, and filter cake is dried, and obtains 4-((3-((tert-butoxy carbonyl Base) amino) phenyl) amino)-2-chloropyrimide-5-Ethyl formate 3.371g, productivity 86%.1H NMR(400MHz, DMSO-d6) δ 10.23 (s, 1H), 9.50 (s, 1H), 8.80 (s, 1H), 7.70 (s, 1H), 7.35 (d, J=8.0 Hz, 1H), 7.29 (t, J=8.0Hz, 1H), 7.23 (d, J=8.0Hz, 1H), 4.38 (q, J=7.2Hz, 2H), 1.49 (s, 9H), 1.36 (t, J=7.2Hz, 3H) .LC-MS:m/z:393.1 (M+H)+.
2.4-((3-((tert-butoxycarbonyl) amino) phenyl) amino)-2-(phenyl amino) pyrimidine-5-Ethyl formate Synthesis
Weigh 4-((3-((tert-butoxycarbonyl) amino) phenyl) amino)-2-chloropyrimide-5-Ethyl formate (1.960g, 5mmol), aniline (0.558g, 6mmol) is in 100mL single port flask, addition 40mL acetonitrile dissolving, liter Temperature backflow 3h.TLC follows the tracks of raw material and converts, and is cooled to room temperature, sucking filtration, washing, and filter cake is dried, obtained 4-((3-((uncle Butoxy carbonyl) amino) phenyl) amino)-2-(phenyl amino) pyrimidine-5-Ethyl formate 1.930g, productivity 86%.1H NMR(400MHz,CDCl3) δ 10.46 (s, 1H), 8.76 (s, 1H), 7.57 (d, J=8.0Hz, 2H), 7.36 (t, J=7.6Hz, 3H), 7.23 (d, J=8.0Hz, 1H), 7.16 (t, J=7.2Hz, 1H), 7.04 (d, J=7.6Hz, 1H), 6.36 (s, 1H), 4.37 (q, J=7.2Hz, 2H), 1.54 (s, 9H), 1.41 (t, J=7.2 Hz,3H).LC-MS:m/z:450.4(M+H)+.
The synthesis of 3.4-((3-((tert-butoxycarbonyl) amino) phenyl) amino)-2-(phenyl amino) pyrimidine-5-formic acid
Weigh 4-((3-((tert-butoxycarbonyl) amino) phenyl) amino)-2-(phenyl amino) pyrimidine-5-Ethyl formate (1.796g, 4mmol), in 100mL single port flask, adds 30mL oxolane and dissolves, drip 1M NaOH Solution 10mL, is warming up to 50 DEG C, stirs 3h.TLC tracks to raw material and converts, and is cooled to room temperature, adds Dilute hydrochloric acid regulation pH is to acid, and sucking filtration, washing, filter cake is dried, obtained 4-((3-((tert-butoxycarbonyl) amino) Phenyl) amino)-2-(phenyl amino) pyrimidine-5-formic acid 1.499g, productivity 89%.1H NMR(400MHz, DMSO-d6) δ 10.44 (s, 1H), 9.82 (s, 1H), 9.36 (s, 1H), 8.70 (s, 1H), 7.69 (d, J=8.0 Hz, 2H), 7.63 (s, 1H), 7.43 (s, 1H), 7.26-7.21 (m, 4H), 6.98 (t, J=6.8Hz, 1H), 1.47 (s, 9H) .HRMS (ESI): value of calculation C22H24N5O4(M+H)+422.1828, experiment value 422.1823.
4.3-(5-(methylcarbamoyl-2-(phenyl amino) pyrimidine-4-amino) phenyl) t-butyl carbamate Synthesis
Weigh 4-((3-((tert-butoxycarbonyl) amino) phenyl) amino)-2-(phenyl amino) pyrimidine-5-formic acid (1.263g, 3mmol), DIPEA (0.774g, 6mmol), in 50mL flask, add 10mL dry DMF Dissolve.Separately take HATU (1.368g, 3.6mmol), be dividedly in some parts in above-mentioned reactant liquor, 1h is stirred at room temperature. Add methylamine hydrochloride (0.396g, 6mmol), stirred overnight at room temperature.TLC follows the tracks of raw material and converts, and adds Frozen water, sucking filtration, washing, obtain 3-(5-(methylcarbamoyl-2-(phenyl amino) pyrimidine-4-amino) phenyl) ammonia Base t-butyl formate 0.612g, productivity 47%.1H NMR(400MHz,DMSO-d6)δ11.33(s,1H), 9.59 (s, 1H), 9.31 (s, 1H), 8.64 (s, 1H), 8.50 (d, J=4.8Hz, 1H), 7.68 (d, J=7.8Hz, 2H), 7.62 (s, 1H), 7.49 (br, 1H), 7.23 (t, J=7.8Hz, 2H), 7.19 (d, J=8.0Hz, 1H), 7.15 (d, J=8.0Hz, 1H), 6.96 (t, J=7.2Hz, 1H), 2.79 (d, J=4.4Hz, 3H), 1.47 (s, 9H).LC-MS:m/z:435.2(M+H)+.
5. (3-(3-methyl-2,4-dioxo-7-(phenyl amino)-3,4-dihydro-pyrimidin also [4,5-d] pyrimidine-1 (2H)- Base) phenyl) t-butyl carbamate
Weigh 3-(5-(methylcarbamoyl-2-(phenyl amino) pyrimidine-4-amino) phenyl) the tertiary fourth of carbamic acid Ester (0.434g, 1mmol), K2CO3(0.276g, 2mmol), in 25mL flask, adds 5mL anhydrous four Hydrogen furan dissolves, and temperature rising reflux is overnight.TLC follows the tracks of raw material and converts, and is cooled to room temperature, adds frozen water, EA Extraction, collected organic layer, rotary evaporation remove solvent, thick product through silica gel column chromatography separate (EA/PE=2:1, V/v), 193mg, productivity 42% are obtained.1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.92(s, 1H), 7.66 (s, 1H), 7.52 (d, J=8.0Hz, 1H), 7.45 (t, J=8.0Hz, 1H), 7.36-7.32 (m, 2H), 7.02 (d, J=8.0Hz, 3H), 6.90 (d, J=7.6Hz, 1H), 3.29 (s, 3H), 1.45 (s, 9H). HRMS (ESI): value of calculation C24H25N6O4(M+H)+461.1937, experiment value 461.1939.
6.1-(3-aminophenyl)-3-methyl-7-(phenyl amino) pyrimido [4,5-d] pyrimidine-2,4 (1H, 3H)-diketone Synthesis
Weigh (3-(3-methyl-2,4-dioxo-7-(phenyl amino)-3,4-dihydro-pyrimidin also [4,5-d] pyrimidine -1 (2H)-yl) phenyl) t-butyl carbamate (0.193g, 0.42mmol) in 50mL flask, add 5mL Dichloromethane dissolves, slowly dropping 1mL trifluoroacetic acid, stirred overnight at room temperature.TLC follows the tracks of raw material and converts, Add saturated NaHCO3Solution neutralizes, and dichloromethane extracts, collected organic layer, and rotary evaporation removes solvent, Obtain 1-(3-aminophenyl)-3-methyl-7-(phenyl amino) pyrimido [4,5-d] pyrimidine-2,4 (1H, 3H)-diketone 0.136g, productivity 90%, product is the most purified is directly used in the next step.
7.N-(3-(3-methyl-2,4-dioxo-7-(phenyl amino)-3,4-dihydro-pyrimidin also [4,5-d] pyrimidine -1 (2H)-yl) phenyl) synthesis (compound 001) of acrylamide
Weigh 1-(3-aminophenyl)-3-methyl-7-(phenyl amino) pyrimido [4,5-d] pyrimidine-2,4 (1H, 3H)- Diketone (0.136g, 0.38mmol), DIPEA (0.097g, 0.76mmol) and 25mL flask, add 3mL Dichloromethane dissolves, and stirs 15 minutes under ice bath.Separately take acryloyl chloride (0.041g, 0.46mmol) and be dissolved in 1 ML dichloromethane, is added drop-wise in above-mentioned reactant liquor, stirred overnight at room temperature.TLC follows the tracks of raw material and converts, and is spin-dried for, Crude product separates (EA/PE=1.5:1, v/v) through silica gel column chromatography, obtains N-(3-(3-methyl-2,4-dioxo-7-(benzene Base amino)-3,4-dihydro-pyrimidin also [4,5-d] pyrimidine-1 (2H)-yl) phenyl) acrylamide 83mg, productivity 53%.1H NMR(400MHz,DMSO-d6) δ 10.38 (s, 1H), 8.93 (s, 1H), 7.82 (d, J=3.2Hz, 1H), 7.80 (s, 1H), 7.53 (t, J=8.0Hz, 1H), 7.33-7.29 (m, 2H), 7.15 (d, J=7.8Hz, 1H), 7.02-6.96 (m, 2H), 6.89 (t, J=6.0Hz, 1H), 6.44 (dd, J=17.2Hz, J=10.4Hz, 1H), 6.26 (dd, J=17.2Hz, J=1.6Hz, 1H), 5.76 (dd, J=10.4Hz, J=2.0Hz, 1H), 3.30 (s, 3H) .HRMS (ESI): value of calculation C22H19N6O3(M+H)+415.1519, experiment value 415.1512.
Following compound (compound 002-010) all obtains according to the method synthesis of above-mentioned steps a-g:
((7-((4-methoxyphenyl) amino)-3-methyl-2,4-dioxo-3,4-dihydro-pyrimidin also [4,5-d] is phonetic for 3-for N- Pyridine-1 (2H)) phenyl) acrylamide (compound 002)
1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),10.23(s,1H),8.88(s,1H), 7.82 (d, J=8.0Hz, 1H), 7.79 (s, 1H), 7.53 (t, J=8.0Hz, 1H), 7.22 (d, J=7.6Hz, 2H), 7.12 (d, J=8.0Hz, 1H), 6.56 (d, J=7.6Hz, 2H), 6.44 (dd, J=16.8Hz, J= 10.0Hz, 1H), 6.24 (dd, J=16.8Hz, J=1.6Hz, 1H), 5.76 (dd, J=10.0Hz, J= 1.6Hz, 1H), 3.65 (s, 3H), 3.29 (s, 3H) .HRMS (ESI) value of calculation C23H21N6O4[M+H]+445.1624, experiment value 445.1627.
((7-((2-methoxyphenyl) amino)-3-methyl-2,4-dioxo-3,4-dihydro-pyrimidin also [4,5-d] is phonetic for 3-for N- Pyridine-1 (2H)) phenyl) acrylamide (compound 003)
1H NMR(400MHz,DMSO-d6)δ10.34(s,1H),8.89(s,1H),8.68(s,1H), 7.80 (s, 1H), 7.76 (d, J=8.0Hz, 1H), 7.50 (t, J=8.0Hz, 1H), 7.41 (d, J=8.0Hz, 1H), 7.12 (d, J=7.6Hz, 1H), 6.95 (d, J=7.6Hz, 2H), 6.44 (dd, J=16.8Hz, J= 10.0Hz, 1H), 6.25 (dd, J=17.2Hz, J=2.0Hz, 1H), 5.76 (dd, J=10.4Hz, J= 2.0Hz, 1H), 3.78 (s, 3H), 3.29 (s, 3H) .HRMS (ESI) value of calculation C23H21N6O4[M+H]+445.1624, experiment value 445.1631.
((-3-methyl-2,4-dioxo-3,4-dihydro-pyrimidin is also for 7-((2-methoxyl group-4-morphlinophenyl) amino) for 3-for N- [4,5-d] pyrimidine-1 (2H)) phenyl) acrylamide (compound 004)
1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.80(s,1H),8.44(s,1H),8.03 (s, 1H), 7.85 (d, J=8.0Hz, 1H), 7.69 (t, J=2.0Hz, 1H), 7.52 (t, J=8.4Hz, 1H), (7.31 d, J=8.8Hz, 1H), 7.09 (d, J=7.6Hz, 1H), 6.57 (d, J=2.0Hz, 1H), 6.45 (dd, J=16.8Hz, J=10.0Hz, 1H), 6.26 (dd, J=17.2Hz, J=2.0Hz, 1H), 5.77 (dd, J =10.4Hz, J=2.0Hz, 1H), 3.77 (s, 3H), 3.58-3.54 (m, 4H), 3.06-2.99 (m, 4H), 2.05 (s, 3H) .HRMS (ESI) value of calculation C27H28N7O5[M+H]+528.1995, experiment value 528.1993.
((7-((2-methoxyl group-4-thiomorpholine is for phenyl) amino)-3-methyl-2,4-dioxo-3,4-dihydro is phonetic for 3-for N- Pyridine also [4,5-d] pyrimidine-1 (2H)) phenyl) acrylamide (compound 005)
1H NMR(400MHz,DMSO-d6)δ10.35(s,1H),8.84(s,1H),8.60(s,1H),8.85 (s, 1H), 7.70 (s, 1H), 7.48 (t, J=8.0Hz, 1H), 7.17 (d, J=8.4Hz, 1H), 7.09 (d, J= 7.6Hz, 1H), 6.44 (dd, J=17.2Hz, J=10.0Hz, 1H), 6.27 (dd, J=16.8Hz, J=2.0 Hz, 1H), 5.97 (s, 1H), 5.77 (dd, J=10.0Hz, J=1.6Hz, 1H), 3.75 (s, 3H), 3.42-3.39 (m, 4H), 3.28 (s, 3H), 2.66-2.63 (m, 4H) .HRMS (ESI) value of calculation C27H28N7O4S[M+H]+546.1923, experiment value 546.1924.
N-(3-(7-((4-(4-acetyl group-3-)-2-methoxyphenyl) amino)-3-methyl-2,4-dioxo-3,4-dihydro Pyrimido [4,5-d] pyrimidine-1 (2H)) phenyl) acrylamide (compound 006)
1H NMR(400MHz,DMSO-d6)δ10.34(s,1H),8.83(s,1H),8.59(s,1H),7.86 (s, 1H), 7.69 (s, 1H), 7.48 (t, J=8.4Hz, 1H), 7.17 (d, J=8.8Hz, 1H), 7.09 (d, J= 8.0Hz, 1H), 6.55 (s, 1H), 6.44 (dd, J=17.2Hz, J=10.4Hz, 1H), 6.26 (dd, J= 17.2Hz, J=1.6Hz, 1H), 5.99 (s, 1H), 5.76 (dd, J=10.0Hz, J=1.2Hz, 1H), 3.76(s,3H),3.58-3.54(m,4H),3.28(s,3H),3.05-2.99(m,4H),2.05(s,3H). HRMS (ESI) value of calculation C29H31N8O5[M+H]+571.2417, experiment value 571.2417.
N-(3-(7-((4-(4-(dimethylamino) piperidines)-2-methoxyphenyl) amino)-3-methyl-2,4-dioxo -3,4-dihydro-pyrimidin also [4,5-d] pyrimidine-1 (2H)) phenyl) acrylamide (compound 007)
1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.83(s,1H),8.54(s, 1H), 7.88-7.86 (m, 1H), 7.70 (s, 1H), 7.47 (t, J=8.0Hz, 1H), 7.16 (d, J=8.4Hz, 1H), 7.08 (d, J=8.0Hz, 1H), 6.50-6.43 (m, 2H), 6.26 (dd, J=16.8Hz, J=1.6Hz, 1H), 5.98 (s, 1H), 5.76 (dd, J=10.0Hz, J=1.2Hz, 1H), 3.75 (s, 3H), 3.59-3.58 (m,2H),3.28(s,3H),2.60-2.55(m,2H),2.22(s,6H),1.82-1.79(m,2H),1.47-1.40 (m, 2H) .HRMS (ESI) value of calculation C30H35N8O4[M+H]+571.2781, experiment value 571.2780.
N-(3-(7-((4-(4-acetylpiperazine base)-2-methoxyphenyl) amino)-3-ethyl-2,4-dioxo-3,4-two Hydrogen pyrimido [4,5-d] pyrimidine-1 (2H)) phenyl) acrylamide (compound 008)
1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.85(s,1H),8.64(s,1H), 7.88 (d, J=4.0Hz, 1H), 7.69 (s, 1H), 7.49 (t, J=8.0Hz, 1H), 7.18 (d, J=8.4Hz, 1H), 7.12 (d, J=7.6Hz, 1H), 6.55 (s, 1H), 6.46 (dd, J=17.2Hz, J=10.0Hz, 1H), 6.27 (dd, J=16.8Hz, J=1.6Hz, 1H), 6.00-5.97 (m, 1H), 5.78 (dd, J=10.0Hz, J =2.0Hz, 1H), 3.94 (q, J=6.4Hz, 2H), 3.77 (s, 3H), 3.58-3.54 (m, 4H), 3.05-2.98 (m, 4H), 2.06 (s, 3H), 1.19 (t, J=6.8Hz, 3H) .HRMS (ESI) value of calculation C30H33N8O5 [M+H]+585.2574, experiment value 585.2572.
N-(3-(7-((4-(4-acetylpiperazine base)-2-methoxyphenyl) amino)-3-isopropyl-2,4-dioxo-3,4- Dihydro-pyrimidin also [4,5-d] pyrimidine-1 (2H)) phenyl) acrylamide (compound 009)
1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),8.83(s,1H),8.61(s,1H), 7.86 (d, J=8.0Hz, 1H), 7.67 (s, 1H), 7.48 (t, J=8.0Hz, 1H), 7.17 (d, J=7.6Hz, 1H), 7.10 (d, J=7.6Hz, 1H), 6.55 (s, 1H), 6.45 (dd, J=16.8Hz, J=10.0Hz, 1H), 6.27 (dd, J=16.8Hz, J=1.6Hz, 1H), 6.00-5.97 (m, 1H), 5.78 (dd, J=10.0Hz, J =2.0Hz, 1H), 5.15-5.11 (m, 1H), 3.76 (s, 3H), 3.57-3.54 (m, 4H), 3.05-2.97 (m, 4H), 2.05 (s, 3H), 1.44 (d, J=6.4Hz, 6H) .HRMS (ESI) value of calculation C31H35N8O5 [M+H]+599.2730, experiment value 599.2731.
N-(3-(7-((4-(4-Acetylpiperazine-1-base)-2-methoxyphenyl) amino)-3-benzyl-2,4-dioxo -3,4-dihydro-pyrimidin also [4,5-d] pyrimidine-1 (2H)-yl) phenyl) acrylamide (compound 010)
1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.87(s,1H),8.69(s,1H),7.87 (d, J=5.6Hz, 1H), 7.68 (s, 1H), 7.48 (t, J=8.0Hz, 1H), 7.38 (d, J=6.8Hz, 2H), 7.33 (t, J=7.2Hz, 2H), 7.26 (t, J=7.2Hz, 1H), 7.17 (d, J=8.0Hz, 1H), 7.12 (d, J=7.8Hz, 1H), 6.55 (s, 1H), 6.44 (dd, J=16.8Hz, J=10.0Hz, 1H), 6.26 (dd, J =17.2Hz, J=2.0Hz, 1H), 5.98 (s, 1H), 5.77 (dd, J=10.0Hz, J=1.6Hz, 1H), 5.09(s,2H),3.76(s,3H),3.58-3.55(m,4H),3.05-2.97(m,4H),2.05(s,3H). HRMS (ESI) value of calculation C35H35N8O5[M+H]+647.2730, experiment value 647.2735.
The concrete synthetic method of compound 011 is as follows:
Reagent and condition: (a)-(e) is as previously mentioned；(f) N, N, N'-trimethyl ethylenediamine, K2CO3,DMSO； (n)Zn,NH4Cl,CH3OH；(o) acyl chlorides, Et3N,CH2Cl2, 0 DEG C of room temperature.
7-((4-((2-(dimethylamino) ethyl) (methyl) amino-2-methoxyl group-5-nitrobenzophenone) amino)-3-methyl The synthesis of-1-phenyl pyrimidine also [4,5-d] pyrimidine-2,4 (1H, 3H)-diketone
Weigh 7-((4-fluoro-2-methoxyl group-5-nitrobenzophenone) amino)-3-methyl isophthalic acid-phenyl pyrimidine also [4,5-d] pyrimidine -2,4 (1H, 3H)-diketone (438mg, 1mmol), K2CO3(207mg, 1.5mmol), N, N, N'-front three Base ethylenediamine (153mg, 1.5mmol), in 10mL single port flask, adds 4mL DMSO and dissolves, be warming up to 90 DEG C, stir 4h.TLC tracks to raw material and converts, and is cooled to room temperature, adds frozen water, and ethyl acetate extracts, Collecting organic facies, be spin-dried for, crude product separates (DCM/CH through silica gel column chromatography3OH=20:1, v/v), 7-((4-((2-(dimethylamino) ethyl) (methyl) amino-2-methoxyl group-5-nitrobenzophenone) amino)-3-methyl isophthalic acid- Phenyl pyrimidine also [4,5-d] pyrimidine-2,4 (1H, 3H)-diketone 358mg, productivity 69%.1H NMR(400MHz, CDCl3)δ8.97(s,1H),7.88(s,1H),7.63(s,1H),7.46-7.43(m,3H),7.19-7.16(m, 2H), 6.52 (s, 1H), 3.83 (s, 3H), 3.39 (s, 3H), 3.08 (t, J=6.4Hz, 2H), 2.71 (s, 3H), 2.41 (t, J=6.8Hz, 2H), 2.17 (s, 6H) .LC-MS:m/z:521.3 (M+H)+.
7-((5-amino-4-((2-(dimethylamino) ethyl) (methyl) amino)-2-methoxyphenyl) amino)-3-first The synthesis of base-1-phenyl pyrimidine also [4,5-d] pyrimidine-2,4 (1H, 3H)-diketone
Weigh 7-((4-((2-(dimethylamino) ethyl) (methyl) amino-2-methoxyl group-5-nitrobenzophenone) amino)-3- Methyl isophthalic acid-phenyl pyrimidine also [4,5-d] pyrimidine-2,4 (1H, 3H)-diketone (312mg, 0.6mmol) is mono-in 50mL Mouth flask, adds 8mL methanol and dissolves, instillation 1mL saturated ammonium chloride solution, and addition zinc powder (390mg, 6 Mmol) stirred overnight at room temperature.TLC tracks to raw material and converts, and filters out zinc powder, and filtrate is spin-dried for, crude product warp Silica gel column chromatography separates (DCM/CH3OH=15:1, v/v), obtain 7-((5-amino-4-((2-(dimethylamino) second Base) (methyl) amino)-2-methoxyphenyl) amino)-3-methyl isophthalic acid-phenyl pyrimidine also [4,5-d] pyrimidine -2,4 (1H, 3H)-diketone 180mg, productivity 61%.1H NMR(400MHz,CDCl3)δ8.94(s,1H), 8.11 (s, 1H), 7.60 (t, J=7.6Hz, 2H), 7.52 (t, J=7.2Hz, 1H), 7.33 (d, J=7.6Hz, 2H), 6.89 (s, 1H), 6.51 (s, 1H), 3.70 (s, 3H), 3.40 (s, 3H), 2.81 (t, J=6.4Hz, 2H), 2.52 (s, 3H), 2.29 (t, J=6.8Hz, 2H), 2.19 (s, 6H) .LC-MS:m/z:491.3 (M+H)+.
N-(2-((2-(dimethylamino) ethyl) (methyl) amino)-4-methoxyl group-5-((6-methyl-5,7-dioxo-8- Phenyl-5,6,7,8-tetrahydropyrimidine also [4,5-d] pyrimidine-2-base) amino) phenyl) synthesis (compound of acrylamide 011)
Weigh 7-((5-amino-4-((2-(dimethylamino) ethyl) (methyl) amino)-2-methoxyphenyl) ammonia Base)-3-methyl isophthalic acid-phenyl pyrimidine also [4,5-d] pyrimidine-2,4 (1H, 3H)-diketone (147mg, 0.3mmol), DIPEA (77mg, 0.6mmol), in 25mL flask, adds 3mL dichloromethane and dissolves, stir under ice bath 15 minutes.Separately take acryloyl chloride (33mg, 0.36mmol) and be dissolved in 1mL dichloromethane, be added drop-wise to above-mentioned instead Answer in liquid, stirred overnight at room temperature.TLC follows the tracks of raw material and converts, and is spin-dried for, and crude product separates through silica gel column chromatography (DCM/CH3OH=15:1, v/v), obtain N-(2-((2-(dimethylamino) ethyl) (methyl) amino)-4-methoxyl group -5-((6-methyl-5,7-dioxo-8-phenyl-5,6,7,8-tetrahydropyrimidine also [4,5-d] pyrimidine-2-base) amino) phenyl) Acrylamide 80mg, productivity 49%.1H NMR(400MHz,CDCl3)δ9.36(s,1H),9.08(s, 1H), 7.88 (s, 1H), 7.50-7.46 (m, 5H), 6.58 (dd, J=17.2Hz, J=10.0Hz, 1H), 6.48 (dd, J=17.2Hz, J=1.6Hz, 1H), 5.83 (dd, J=10.0Hz, J=2.0Hz, 1H), 3.83 (s, 3H), 3.48 (s, 3H), 3.23 (t, J=6.0Hz, 2H), 3.06 (t, J=6.4Hz, 2H), 2.77 (s, 6H), 2.68 (s, 3H) .HRMS (ESI): value of calculation C28H33N8O4(M+H)+545.2625, experiment value 545.2630。
Embodiment 2. biological activity test
The extracorporeal extracorporeal suppression of EGFR kinase activity is tested and is carried out as follows by the compound that the present invention provides:
Vitro enzyme activity analysis: wild type and various saltant type (T790M, L858/T790M) EGFR all purchase From in Invitrogen.It is provided with from 5.1 × 10 for all of compound to be tested-11Mol/L to 1.0 × 10-610 Concentraton gradient of mol/L.
Different kinase whose concentration are determined by optimization experiment, and corresponding concentration is: EGFR (PV3872, Invitrogen)0.287μg/μL,EGFR-T790M(PV4803,Invitrogen)0.174μg/μL, EGFR-L858R/T790M(PV4879,Invitrogen)0.055μg/μL.Compound in DMSO from 5.1x10-9M to 1x10-4M dilutes three times.4 μ L compounds are dissolved in 96 μ L water, obtain the compound solution of 4x. 40 μMs of ATP are dissolved in 1.33x kinase buffer liquid, and kinases/peptide mixer comprises 2x kinases, 4 μMs of trorsine 14 peptides It is ready to stand-by.10 μ L kinase reactions include 2.5 μ L compound solutions, 5 μ L kinases/peptide mixer, 2.5 μ L ATP solution.5 μ L Phosphorylated Peptide solution replace kinases/peptide mixer to be used as 100% phosphorylation comparison.2.5μL 1.33x kinase buffer liquid replaces ATP solution to be used as 100% suppression comparison, and 2.5 μ L 4%DMSO replace chemical combination Thing solution is used as 0% suppression comparison.At room temperature cultivate after in plate, solution is sufficiently mixed 1.5 hours.Every hole adds Continue at room temperature to cultivate 1 hour after entering 5 μ L Development Solution, non-phosphorylating peptide at this moment between Interior cleaved.Finally, add 5 μ L and terminate preparation (Stop Reagent) end reaction.Orifice plate is used EnVisionMultilabel Reader (Perkin Elmer) measures.Experimental data uses GraphPad Prism version 4.0 calculates.Experiment is all repeated 3 times above every time.
Cell proliferation and growth inhibited analysis: H1975 (non-small cell lung cancer cell, EGFRL858R/T790M)、 A431 (non-small cell lung cancer cell, EGFR wild type), cell all obtains from ATCC.Cell-proliferation activity MTS analytic process is used to be estimated.Cell exposes 72 hours under processing conditions, and each cell line is tested every time The cell number used is adjusted according to absorbance (absorbance at 490nm is 1.3-2.2).For treating Test compound is provided with 6 Concentraton gradient (0.1nM-10 μM), and each concentration value at least uses 6 groups to put down Row comparison.
H1975, A431 cell is cultivated in corresponding culture medium, and cell at least passes on twice after recovery, It is subsequently used for experiment to use.The cell of logarithmic (log) phase is by trypsin acting settling flux in the medium.H1975 (every hole 1000 cell), A431 (every hole 2000 cell) are seeded in 96 orifice plates, volume 100 μ L；Arrange 6 Organize parallel and 7 row.Orifice plate is put in the incubator of 37 DEG C of 5% carbon dioxide overnight.Compound is dissolved in DMSO, compound concentration is 10 μMs every liter, compound concentration progressively dilutes compound subsequently that obtain dense Degree is respectively 10 μMs every liter, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM.2μL Compound solution is added in the culture medium of 998 μ L, and mixture is adequately mixed.The mixture of 100 μ L adds In 96 orifice plates.2 μ L DMSO replace compound solution to be used as 0% suppression comparison.After cultivating 68 hours, add Enter 20 μ L MTT (5mg/mL).4 as a child, abandons supernatant and adds 150 μ L DMSO.Shake 10 After minute, orifice plate Synergy HT (Bio TeK) (OD490) reads data.Data use GraphPad Prism version 4.0 calculates, IC50It is worth and is adjusted by the nonlinear regression model (NLRM) of using dosage response curve Whole obtain.
Test result is as shown in table 1 below.
Table 1
*: > 100 represent substantially can't detect the inhibitory action to cell.
Discuss:
Inventor, through extensively in-depth study, designs and synthesizes and has obtained a series of having no what document was reported Pyrimido [4,5-d] pyrimidine-2,4 (1H, 3H)-cyclohexadione compounds, the compound obtained has been carried out molecular level With the active testing of cellular level, obtain a collection of can Selective depression EGFR T790M sudden change compound. Present inventors have further discovered that, the compound of the present invention is to EGFR saltant type cancerous cell (H1975) and EGFR The Proliferation Ability difference of ability of wild type cancerous cell (A431) swashs higher than to mutant egf R and Wild type EGFR Inhibition of enzyme activity difference of ability, thus point out the compound of the present invention to have more preferable difference toxicity in vivo, have It is likely to become Selective depression T790M sudden change, overcomes the third generation EGFR targeted drug of clinical drug-resistant, or Person is as obtaining activity more preferably and/or the basis of difference toxicity more preferably compound through modifying further.
The all documents mentioned in the present invention are incorporated as reference the most in this application, just as each document It is individually recited as with reference to like that.In addition, it is to be understood that after the above-mentioned teachings having read the present invention, The present invention can be made various changes or modifications by those skilled in the art, and these equivalent form of values fall within this Shen equally Please appended claims limited range.

Claims (10)

1. the compound shown in formula I or its pharmaceutically acceptable salt:
In formula, A is phenyl ring, five yuan or hexa-member heterocycle, C3-C8Cycloalkyl；
R1It is each independently selected from hydrogen, halogen, C1-C3Alkoxyl, C1-C3Alkyl, C1-C4Alkylamidoalkyl, Substituted piperazinyl, substituted homopiperazine base, substituted morpholinyl, substituted thio-morpholinyl, 4-N-first Base piperazinyl, 4-N-acetylpiperazinyl, 4-N, N-lupetidine base, substituted piperidyl ,-NRaRb, Wherein, RaAnd RbAlkyl can be independently selected from and containing azanyl；
R2It is each independently selected from following group:
R3Selected from lower group: hydrogen, C1-C10Alkyl, substituted C1-C10Alkyl, C2-C6Alkenyl, C2-C6Alkynes Base, optionally substituted C3-C8Cycloalkyl, optionally substituted aryl, optionally substituted benzyl, optionally substituted Heterocyclic radical, optionally substituted aromatic heterocyclic；
B is selected from lower group:
M is the integer of 0-7.
2. compound as claimed in claim 1 or its pharmaceutically acceptable salt, it is characterised in that described Compound is as shown in formula II:
In formula, B, R1、R2、R3As defined in claim 1；
M is the integer of 0-5.
3. compound as claimed in claim 2 or its pharmaceutically acceptable salt, it is characterised in that described Compound is as shown in general formula III:
In formula,
R2It is selected fromOr
R3Selected from C1-C6Alkyl, preferably methyl or isopropyl；Or the substituted C of phenyl1-C6Alkyl, preferably benzyl Base；
R4、R5、R6、R7And R8It is independently selected from lower group:
4. compound as claimed in claim 3 or its pharmaceutically acceptable salt, it is characterised in that R5With R8For H；Or, R5、R7And R8For H.
5. selected from compound or its pharmaceutically acceptable salt of lower group:
6. a pharmaceutical composition, described pharmaceutical composition contains the change according to any one of claim 1-5 Compound or its pharmaceutically acceptable salt, and pharmaceutically acceptable carrier or excipient.
7. the compound according to any one of claim 1-5 is in preparation treatment or the disease of prevention EGFR mediation Purposes in disease, or the medicine of suppression EGFR.
8. purposes as claimed in claim 7, it is characterised in that the disease of described EGFR mediation is cancer.
9. purposes as claimed in claim 8, it is characterised in that described cancer is selected from lower group: non-small cell Pulmonary carcinoma, small cell lung cancer, adenocarcinoma of lung, lung squamous cancer, breast carcinoma, carcinoma of prostate, neurogliocytoma, Ovarian cancer, G. cephalantha, cervical cancer, the esophageal carcinoma, hepatocarcinoma, renal carcinoma, cancer of pancreas, colon cancer, skin Cancer, leukemia, lymphoma, gastric cancer, multiple bone marrow cancer and solid tumor.
10. utilize the compounds for treating according to any one of claim 1-5 or the disease of prevention EGFR mediation Method.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112321814A (en) * 2020-12-30 2021-02-05 广州初曲科技有限公司 Preparation and application of gefitinib idebenone conjugate
JP2022546375A (en) * 2019-08-23 2022-11-04 北京泰徳製薬股▲フン▼有限公司 Compounds that inhibit EGFR and ALK to inhibit their degradation

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105985342B (en) * 2015-02-06 2018-07-24 华东理工大学 As the pyrimido-pyrimidine derovatives of EGFR inhibitor and its application
CN107129506B (en) * 2016-02-26 2019-10-22 华东理工大学 As pyrimido [4,5-d] [1,3] oxazines -2- ketone derivatives of EGFR inhibitor and its application
CN108727382B (en) * 2017-04-19 2022-07-19 华东理工大学 Heterocyclic compounds as BTK inhibitors and uses thereof
US10449186B2 (en) * 2017-06-21 2019-10-22 University Of Kentucky Research Foundation Phenylethynyl-substituted benzenes and heterocycles for the treatment of cancer
US20240124464A1 (en) * 2020-12-02 2024-04-18 Impact Therapeutics (Shanghai), Inc Substituted fused bicyclic compound as kinase inhibitor and use thereof
WO2023239165A1 (en) * 2022-06-08 2023-12-14 한국화학연구원 Phenylsulfonamide derivatives, preparation method therefor, and pharmaceutical composition for prevention or treatment of cancer containing same as active ingredient

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010067888A1 (en) * 2008-12-12 2010-06-17 Banyu Pharmaceutical Co.,Ltd. Dihydropyrimidopyrimidine derivatives
CN102816162A (en) * 2011-06-10 2012-12-12 中国科学院广州生物医药与健康研究院 Pyrimidine, pyrimidone compound, and medical compound and application thereof
WO2015006492A1 (en) * 2013-07-09 2015-01-15 Dana-Farber Cancer Institute, Inc. Kinase inhibitors for the treatment of disease

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105985342B (en) * 2015-02-06 2018-07-24 华东理工大学 As the pyrimido-pyrimidine derovatives of EGFR inhibitor and its application

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010067888A1 (en) * 2008-12-12 2010-06-17 Banyu Pharmaceutical Co.,Ltd. Dihydropyrimidopyrimidine derivatives
CN102816162A (en) * 2011-06-10 2012-12-12 中国科学院广州生物医药与健康研究院 Pyrimidine, pyrimidone compound, and medical compound and application thereof
WO2015006492A1 (en) * 2013-07-09 2015-01-15 Dana-Farber Cancer Institute, Inc. Kinase inhibitors for the treatment of disease

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
任晓岚等: "表皮生长因子受体(EGFR)酪氨酸激酶抑制剂的研究进展", 《中国新药杂志》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2022546375A (en) * 2019-08-23 2022-11-04 北京泰徳製薬股▲フン▼有限公司 Compounds that inhibit EGFR and ALK to inhibit their degradation
CN112321814A (en) * 2020-12-30 2021-02-05 广州初曲科技有限公司 Preparation and application of gefitinib idebenone conjugate
CN112321814B (en) * 2020-12-30 2021-03-23 广州初曲科技有限公司 Preparation and application of gefitinib idebenone conjugate

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