CN104140426B - Pyrimidoimidazole compound and pharmaceutical composition and preparation method and use thereof - Google Patents

Pyrimidoimidazole compound and pharmaceutical composition and preparation method and use thereof Download PDF

Info

Publication number
CN104140426B
CN104140426B CN201310165742.5A CN201310165742A CN104140426B CN 104140426 B CN104140426 B CN 104140426B CN 201310165742 A CN201310165742 A CN 201310165742A CN 104140426 B CN104140426 B CN 104140426B
Authority
CN
China
Prior art keywords
hydrogen
compound
base
formula
benzimidazole
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201310165742.5A
Other languages
Chinese (zh)
Other versions
CN104140426A (en
Inventor
樊兴
秦继红
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Yidian Medical Science and Technology Development Co., Ltd.
Original Assignee
SHANGHAI HUILUN TECHNOLOGY Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by SHANGHAI HUILUN TECHNOLOGY Co Ltd filed Critical SHANGHAI HUILUN TECHNOLOGY Co Ltd
Priority to CN201310165742.5A priority Critical patent/CN104140426B/en
Publication of CN104140426A publication Critical patent/CN104140426A/en
Application granted granted Critical
Publication of CN104140426B publication Critical patent/CN104140426B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention relates to a pyrimidoimidazole derivative, a preparation method and medical use thereof, and particularly relates to a new pyrimidoimidazole derivative as shown in a general formula (I) or formula (II), a preparation method thereof, a pharmaceutical composition containing the new pyrimidoimidazole derivative, and use thereof as a therapeutic agent in particular as a poly (ADP(adenosine diphosphate)-ribose) polymerase (PARP) inhibitor.

Description

Pyrimido glyoxaline compound, its pharmaceutical composition and its production and use
Technical field
The present invention relates to pyrimido imidazole derivative, its preparation method and the pharmaceutical composition containing this derivant, And its purposes as therapeutic agent with as poly- (adp- ribose) polymerase (parp) inhibitor.
Background technology
Chemotherapeutics and ionizing radiation treatment are two kinds of common methods for the treatment of cancer.Both Therapeutic Method all can induce Dna is single-stranded and/or double-strand break and then produce cytotoxic effect, and target tumor is due to chromosome damage thus death. Important results as response dna damage signal are that cell cycle regulating site signal is activated, and its object is to protect Cell does not carry out mitosiss thus avoiding cell injury in the case of dna damage.In most of the cases, tumor cell exists While showing cell cycle regulating site signal defect, there is very high appreciation rate.It can therefore be concluded that in tumor cell There is specific dna repair mechanism, with quick response and the chromosome damage related to breeding regulation can be repaired, so that it is certainly Body survives cytotoxic effect the maintenance survival of some medicines.
In clinical practice, the valid density of chemotherapeutics or treatment radiant intensity can resist these dna repair mechanisms, Ensure the fragmentation effect to target tumor.However, tumor cell can be to treatment by strengthening its dna damage repair mechanism Produce tolerance effect, be allowed to survive from fatal dna damages.In order to overcome the toleration of generation, it usually needs increase The dosage of medicine or raising radiant intensity, this way will have a negative impact to the normal structure near focus, thus Make with serious untoward reaction in therapeutic process, and then increase Operative risk.Meanwhile, ever-increasing toleration will drop Low therapeutic effect, it can therefore be concluded that by the regulation to dna damage signal repair mechanism, can be with tumor cell specific Mode realize the raising of the cytotoxicity to dna damaging agents.
The parps(poly(adp-ribose being characterized with poly- adenosine diphosphate-ribosylating activity) Polymerases), constitute the superfamily of 18 kinds of cell ribozyme nucleuss matter enzymes.This poly- adenosine diphosphate-ribosyl is turned into With catalysis activity and the protein-protein interaction of destination protein can be adjusted, and many basic bioprocesss are adjusted Control, repairs including dna, cell death, Genome stability is also associated.
Parp-1 activity accounts for the 80% of total cell parp activity, and it and the parp-2 the most close with it become jointly Possesses the member repairing dna lesion capability in parp family.The induction apparatuss damaging as dna and signal protein, parp-1 is permissible Quick detection is simultaneously bonded directly to dna injury site, and induced aggregation dna repairs required multiple protein afterwards, and then so that dna is damaged Wound is repaired.When the parp-1 in cell lacks, parp-2 can substitute the reparation that parp-1 realizes dna damage.Research Show, compared with normal cell, expression in solid tumor for the parps albumen generally strengthens.Additionally, repairing dependency basis for dna Because lacking the tumor (as breast tumor and ovarian cancer) of (as brca-1 or brca-2), show extreme to parp-1 inhibitor Sensitivity, this show parp inhibitor as single dose treating this potential use being known as in terms of triple negative breast cancer.Meanwhile, Because dna damage repair mechanism is tumor cell reply chemotherapeutics and the ionizing radiation treatment resistance to main machine the affected of generation System, therefore parp-1 is considered as an Effective target site exploring new cancer treatment method.
The parp inhibitor of early development design is using the nicotiamide of the nad as parp catalytic substrate as template, opens Send out its analog.These inhibitor, as the competitive inhibitor of nad, compete the catalytic site of parp with nad, and then stop poly- The synthesis of (adp- ribose) chain.Parp under not having poly- (adp- ribosylation) to modify cannot disintegrate down from dna injury site, The protein leading to other to participate in repairing be cannot be introduced into injury site, and then repair process can not be executed.Therefore, in cell toxicant Property medicine or radiation in the presence of, the tumor cell that the presence of parp inhibitor makes dna impaired is finally dead.
Additionally, the nad being consumed as parp catalytic substrate, it is requisite during cell synthesis atp, because This, under high parp activity level, intracellular nad level can be remarkably decreased, and then affects the atp level of intracellular.Due to thin The atp content of intracellular is not enough, and cell cannot realize the programmed cell death process of atp dependence, and this special withers can only to turn to necrosis Die process.During necrosis, substantial amounts of inflammatory factor can be released, thus producing toxicity to other organs and tissue Effect.Therefore, parp inhibitor can be used for treating the multiple diseases relevant with this mechanism, including neurodegenerative diseases (such as senile dementia, Huntington chorea, parkinson disease), diabetes, the complication in ischemia or Ischemia-Reperfusion Injury, As myocardial infarction and acute renal failure, blood circulation diseasess, such as septic shock, and diseases associated with inflammation, such as chronic rheumatism etc..
Content of the invention
An object of the present invention is to provide a kind of new pyrimido glyoxaline compound and its derivant, and they Tautomer, enantiomer, diastereomer, raceme and pharmaceutically useful salt, and metabolite and metabolite precursor or Prodrug.
The second object of the present invention is to provide the medicine group using described pyrimido glyoxaline compound as active component Compound.
The third object of the present invention is to provide the preparation method of above-mentioned pyrimido glyoxaline compound.
The fourth object of the present invention is to provide application in medicine for the above-mentioned pyrimido glyoxaline compound.
As the pyrimido glyoxaline compound of first aspect present invention, its be formula (i) or the change shown in formula () Compound:
Wherein: in formula () or formula (), r is hydrogen or halogen；
X, y, z is one of to be nitrogen, remaining be hydrocarbon or x, y, z one of for hydrocarbon, remaining is nitrogen；
M is nitrogen or cr1;
R1 is hydrogen, oxygen, c1-c6Alkyl, methoxyl group, trifluoromethyl and nr2r3；
R2 is hydrogen or c1-c4Alkyl；
R3 is hydrogen or c1-c4Alkyl.
It is further preferred that compound as shown in formula () or formula () for the structure of present invention offer, wherein:
R is hydrogen or fluorine；
X, y, z is one of to be nitrogen, and remaining is hydrocarbon；
M is nitrogen or cr1;
R1 is hydrogen, oxygen, c1-c4Alkyl, methoxyl group, trifluoromethyl and nr2r3；
R2 is hydrogen or c1-c4Alkyl；
R3 is hydrogen or c1-c4Alkyl.
Most preferably, formula () of the present invention or the compound shown in formula () are included for following compound (1)～(16) One of:
Described formula () or formula () compound are enantiomer, in diastereomer, conformer The mixture of any one or arbitrarily both or three.
Described formula () or formula () compound are pharmaceutically acceptable derivant.
Formula () of the present invention or formula () compound can exist as a pharmaceutically acceptable salt form.
Pharmaceutically acceptable salt of the present invention is the hydrochlorate of formula () or formula () compound, sulfate, phosphorus Hydrochlorate, acetate, trifluoroacetate, mesylate, fluoroform sulphonate, tosilate, tartrate, maleate, Fumarate, succinate or malate.
In a preferred embodiment of the invention, the pyrimido glyoxaline compound of described formula () or formula () For 2- (piperazine -1- base) -1 hydrogen-h-benzimidazole-4-carboxamide class compound and its officinal salt.
As the preparation method of the compound shown in the formula () of second aspect present invention, its reaction equation is as follows:
Wherein, r, x, y, z and m are as defined above；Specifically comprise the following steps that
Step 1): substituted 2,3- diamidogen yl benzoic acid methyl ester and carbonyl dimidazoles ring-closure reaction, obtain the 2- oxygen replacing Generation -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate ()；
Step 2): substituted 2- oxo -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate () that step 1) obtains Carry out chlorination reaction with phosphorus oxychloride, obtain chloro- 1 hydrogen of the 2--benzimidazole -4- methyl formate () replacing；
Step 3): in the presence of a base, by step 2) substituted chloro- 1 hydrogen of 2--benzimidazole -4- methyl formate of obtaining () and piperazine carry out nucleophilic substitution, obtain 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate replacing ()；
Step 4): substituted 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate () that step 3) is obtained There is nucleophilic substitution with polysubstituted pyrimidine compound, obtain intermediate ()；
Step 5): the intermediate () that step 4) is obtained occurs catalytic hydrogen reduction nitro, obtains intermediate ()；
Step 6): the intermediate () that step 5) is obtained by with acetic anhydride, trifluoro-acetic anhydride, trimethyl orthoformate or There is ring-closure reaction in Hydrazoic acid,sodium salt, obtain intermediate ()；
Step 7): the intermediate () that step 6) is obtained passes through the aminolysis reaction of ester group in methanolic ammonia solution, Generate the compound shown in formula ().
As the preparation method of the compound shown in the formula () of second aspect present invention, its reaction equation is as follows:
Wherein, r, x, y, z and m are as defined above；Comprise the following steps that
Step (1): substituted 2,3- diamidogen yl benzoic acid methyl ester and carbonyl dimidazoles ring-closure reaction, obtain the 2- oxygen replacing Generation -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate ()；
Step (2): substituted 2- oxo -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate that step (1) obtains () and phosphorus oxychloride carry out chlorination reaction, obtain chloro- 1 hydrogen of the 2--benzimidazole -4- methyl formate () replacing；
Step (3): in the presence of a base, substituted chloro- 1 hydrogen of 2--benzimidazole -4- methyl formate that step (2) is obtained () and piperazine carry out nucleophilic substitution, obtain 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate replacing ()；
Step (4): substituted 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate that step (3) is obtained There is nucleophilic substitution with polysubstituted pyrimidine compound in (), obtain intermediate ()；
Step (5): the intermediate () that step (4) is obtained occurs catalytic hydrogen reduction nitro, obtains intermediate ()；
Step (6): the intermediate () that step (5) is obtained by with acetic anhydride, trifluoro-acetic anhydride, trimethyl orthoformate Or Hydrazoic acid,sodium salt occurs ring-closure reaction, obtain intermediate ()；
Step (7): the intermediate () that step (6) is obtained passes through to occur the amine solution of ester group anti-in methanolic ammonia solution Should, generate the compound shown in formula ().
As the Pharmaceutical composition of third aspect present invention, comprise to constitute the formula () of the therapeutically effective amount of active component Or formula () compound and one or more medicinal carrier substance and/or diluent.Or comprise to constitute controlling of active component Treat formula () or formula () compound and pharmaceutically acceptable carrier, excipient or the diluent of effective dose.
As the Pharmaceutical composition of third aspect present invention, comprise to constitute the formula () of the therapeutically effective amount of active component Or the pharmaceutically acceptable derivant of formula () compound and one or more medicinal carrier substance and/or diluent.Or Comprise the formula () of therapeutically effective amount or the pharmaceutically acceptable derivant of formula () compound and the medicine constituting active component Acceptable carrier, excipient or diluent on.
As the Pharmaceutical composition of third aspect present invention, comprise to constitute the formula () of the therapeutically effective amount of active component Or the pharmaceutically acceptable salt of formula () compound and one or more medicinal carrier substance and/or diluent.Or bag Containing constituting the formula () of therapeutically effective amount of active component or the pharmaceutically acceptable salt of formula () compound and pharmaceutically Acceptable carrier, excipient or diluent.
Described pharmaceutical composition make tablet, capsule, aqueous suspension, Oil suspensions, dispersible powder, Granule, lozenge, Emulsion, syrup, ointment, ointment, suppository or injection.
In described pharmaceutical composition, described formula () or formula () compound exist in a free form.
As the application of fourth aspect present invention, it is wherein described formula () or formula () compound in preparation treatment Application in the disease medicament improving because of parp activity suppression.
As the application of fourth aspect present invention, it is wherein that the pharmacy of described formula () or formula () compound can connect The derivant being subject to applying in the disease medicament that preparation treatment improves because of parp activity suppression.
As the application of fourth aspect present invention, it is wherein the pharmaceutically useful of described formula () or formula () compound Application in the disease medicament that preparation treatment improves because of parp activity suppression for the salt.
As the application of fourth aspect present invention, it is wherein that described pharmaceutical composition is treated because of parp activity suppression in preparation And the application in the disease medicament improving.
The described disease improved because of parp activity suppression is angiopathy, septic shock, ischemic injuries, Nervous toxicity Property, hemorrhagic shock, inflammatory diseasess or multiple sclerosis.
As the application of fourth aspect present invention, it is wherein described formula () or formula () compound is used in preparation Application in the ancillary drug of oncotherapy.
As the application of fourth aspect present invention, it is wherein that the pharmacy of described formula () or formula () compound can connect The derivant being subject to is used for applying in the ancillary drug of oncotherapy in preparation.
As the application of fourth aspect present invention, it is wherein the pharmaceutically useful of described formula () or formula () compound Salt is used for the application in the ancillary drug of oncotherapy in preparation.
As the application of fourth aspect present invention, it is wherein that described pharmaceutical composition is used for the auxiliary of oncotherapy in preparation Application in medicine.
As the application of fourth aspect present invention, it is wherein described formula () or formula () compound is used in preparation Tumor strengthens the application in the medicine of radiotherapy.
As the application of fourth aspect present invention, it is wherein that the pharmacy of described formula () or formula () compound can connect The derivant being subject to strengthens applying in the medicine of radiotherapy in preparation for tumor.
As the application of fourth aspect present invention, it is wherein the pharmaceutically useful of described formula () or formula () compound Salt strengthens the application in the medicine of radiotherapy in preparation for tumor.
As the application of fourth aspect present invention, it is wherein that described pharmaceutical composition strengthens radiotherapy in preparation for tumor Application in medicine.
As the application of fourth aspect present invention, it is wherein described formula () or formula () compound is used in preparation Application in the medicine of chemotherapy of tumors.
As the application of fourth aspect present invention, it is wherein that the pharmacy of described formula () or formula () compound can connect The derivant being subject to is used for applying in the medicine of chemotherapy of tumors in preparation.
As the application of fourth aspect present invention, it is wherein the pharmaceutically useful of described formula () or formula () compound Salt is used for the application in the medicine of chemotherapy of tumors in preparation.
As the application of fourth aspect present invention, it is wherein that described pharmaceutical composition is used for the medicine of chemotherapy of tumors in preparation In application.
As the application of fourth aspect present invention, it is wherein that described formula () or formula () compound lack in preparation Application in the medicine of individuation treatment of cancer that the dependent dna double-strand break of homologous recombination (hr) (dsb) is repaired.
As the application of fourth aspect present invention, it is wherein that the pharmacy of described formula () or formula () compound can connect The derivant being subject to lacks, in preparation, the individuation treatment of cancer that the dependent dna double-strand break (dsb) of homologous recombination (hr) is repaired Medicine in application.
As the application of fourth aspect present invention, it is wherein the pharmaceutically useful of described formula () or formula () compound Salt lacks in the medicine of the individuation treatment of cancer that the dependent dna double-strand break (dsb) of homologous recombination (hr) is repaired in preparation Application.
As the application of fourth aspect present invention, be wherein described pharmaceutical composition preparation lack homologous recombination (hr) according to Application in the medicine of individuation treatment of cancer that the dna double-strand break (dsb) of bad property is repaired.
Preferably, described cancer is the ability of the dsb repairing dna containing one or more by hr with respect to normal Cell and the cancer of cancerous cell that lowers or lose.
Preferably, described cancer is the cancer with brca-1 or brca-2 defect, mutant phenotype.
Preferably, described cancer is breast carcinoma, ovarian cancer, cancer of pancreas or carcinoma of prostate.
In order to check the exposure level for parp enzyme for the compound of present invention offer, using the test of biochemistry level enzymatic activity To determine the various compounds of the present invention activity to parp enzyme.
Parp is a kind of posttranscriptional modification enzyme, and dna damages and can activate this enzyme, and parp catalytic process in vivo is mainly The poly(adp-ribose that a kind of nad relies on) process, its substrate some nucleoprotein mainly including parp, Histone is one of which, and the present invention passes through to measure parp under nad effect to being coated in histone poly in 96 orifice plates (adp-ribose) degree, measures parp activity, correspondingly measures parp activity after the effect of parp inhibitor, thus evaluating such The inhibition level to parp activity for the compound.
Specific embodiment
Unless stated to the contrary, following term in the specification and in the claims has following implications.
In the present invention, term " c1-c6Alkyl " refers to there is straight or branched part the saturation containing 1 to 6 carbon atom Monovalent hydrocarbon.The example of such group includes but is not limited to methyl, ethyl, propyl group, isopropyl, normal-butyl, isobutyl group and tertiary fourth Base.
Term " halogen " and " halo " refer to f, cl, br, i.
" pharmaceutically acceptable salt " represents the reservation biological effectiveness of parent compound and those salt of property.This kind of salt Including:
(1) become salt with acid, obtained, mineral acid bag by the reaction of the free alkali of parent compound and mineral acid or organic acid Include hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, Metaphosphoric acid, sulphuric acid, sulfurous acid and perchloric acid etc., organic acid includes acetic acid, propanoic acid, propylene Acid, oxalic acid, (d) or (l) malic acid, fumaric acid, maleic acid, hydroxy benzoic acid, gamma-hydroxybutyric acid, methoxybenzoic acid, adjacent benzene Dioctyl phthalate, methanesulfonic acid, ethyl sulfonic acid, naphthalene -1- sulfonic acid, naphthalene-2-sulfonic acid, p-methyl benzenesulfonic acid, salicylic acid, tartaric acid, citric acid, breast Acid, mandelic acid, succinic acid or malonic acid etc..
(2) acid proton being present in parent compound is replaced or given birth to organic base ligand compound by metal ion Become salt, metal ion such as alkali metal ion, alkaline-earth metal ions or aluminium ion, organic bases for example ethanolamine, diethanolamine, Triethanolamine, trometamol, n- methylglucosamine etc..
" pharmaceutical composition " refers to one or more of compound in the present invention or its pharmaceutically acceptable salt, molten Agent compound, hydrate or prodrug and other chemical composition, such as pharmaceutically acceptable carrier, mixing.The mesh of pharmaceutical composition Be promote administration to animal process.
" pharmaceutical carrier " refers to the biology not causing obvious zest to organism and not disturbing given compound Non-active ingredient in the pharmaceutical composition of activity and property, such as but not limited to: Calcium Carbonate, calcium phosphate, various sugar (such as breast Sugar, Mannitol etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylate copolymer or methacrylic polymeric Thing, gel, water, Polyethylene Glycol, propylene glycol, ethylene glycol, Oleum Ricini or castor oil hydrogenated or many ethoxy aluminium Oleum Ricini, Semen Sesami Oil, Semen Maydis oil, Oleum Arachidis hypogaeae semen etc..
In aforesaid pharmaceutical composition, in addition to including pharmaceutically acceptable carrier, it is additionally may included in medicine (agent) and learns Upper conventional adjuvant, for example: antibacterial agent, antifungal, antimicrobial, preservative, toner, solubilizing agent, thickening agent, table Face activating agent, chelating agent, protein, aminoacid, fat, saccharide, vitamin, mineral, trace element, sweeting agent, pigment, perfume (or spice) Essence or their combination etc..
The invention discloses a kind of compound and this compound are as the application of poly- (adp- ribose) AG14361, this Skilled person can use for reference present disclosure, be suitably modified technological parameter and realize.Specifically, all similar Replace and change apparent to those skilled in the art, they are considered as including in the present invention.The present invention's Method and application are described by preferred embodiment, and related personnel substantially can be without departing from present invention, spirit With in scope, method described herein and application are modified or suitably change and combine, to realize and to apply skill of the present invention Art.
With reference to embodiment, the present invention be expanded on further:
Preparation embodiment
Embodiment 1
Compound (1): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
The preparation of step 1:2- oxo -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate
In an anhydrous tetrahydrofuran solution (20ml) dissolved with 2,3- diamidogen yl benzoic acid methyl ester (0.8g, 4.8mmol) Middle addition carbonyl dimidazoles (1.56g, 9.6mmol), is warming up to backflow, and reaction cooled down after 8 hours, and removal of solvent under reduced pressure is remaining Thing separates (petroleum ether: ethyl acetate=5:1) through rapid column chromatography and obtains faint yellow solid compound a: 2- oxo -2,3- dihydro - 1 hydrogen-benzimidazole -4- methyl formate (0.3g, yield 33%).ms(esi)m/z:[m+h]+=193.
The preparation of chloro- 1 hydrogen of step 2:2--benzimidazole -4- methyl formate
Compound a (1.1g, 5.7mmol) is added phosphorus oxychloride (8ml), is warming up to backflow, reaction cooled down after 8 hours, Removal of solvent under reduced pressure, residue separates (petroleum ether: ethyl acetate=5:1) through rapid column chromatography and obtains compound as white solid b: Chloro- 1 hydrogen of 2--benzimidazole -4- methyl formate (1.5g, yield 100%).ms(esi)m/z:[m+h]+=211.
The preparation of step 3:2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate
Will be dissolved with compound b(59mg, 0.28mmol) dimethylformamide (5ml) in add piperazine (110mg, 1.12mmol), it is warming up to 100 DEG C, reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue separates (two through rapid column chromatography Chloromethanes: methanol=10:1) obtain compound as white solid c:2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (100mg, yield 100%).ms(esi)m/z:[m+h]+=261.
Step 4:2- (4- (2- amino -3- nitropyridine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- formic acid first The preparation of ester
Will be dissolved with compound c(100mg, 0.41mmol) dimethylformamide (5ml) in add the chloro- 3 nitro -2- ammonia of 4- Yl pyridines (102mg, 0.59mmol) and diisopropyl ethyl amine (95mg, 0.74mmol), room temperature reaction was removed under reduced pressure after 8 hours Solvent, residue separates (dichloromethane: methanol=10:1) through rapid column chromatography and obtains compound as white solid d:2- (4- (2- ammonia Base -3- nitropyridine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate (100mg, yield 54%).ms(esi) m/z:[m+h]+=398.
Step 5:2- (4- (2,3 diamino pyridine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate Preparation
By 10% palladium carbon (20mg) add dissolved with compound d(100mg, 0.26mmol) methanol (10ml) solution in, room temperature Lower hydrogenation 7 hours, filters, and residue separates (dichloromethane: methanol=5:1) through rapid column chromatography and obtains yellow solid compound E:2- (4- (2,3- diamino-pyridine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate (88mg, yield 96%).ms(esi)m/z:[m+h]+=353.
Step 6:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- base) -1 The preparation of hydrogen-benzimidazole -4- methyl formate
At one dissolved with compound e(88mg, 0.25mmol) anhydrous tetrahydrofuran solution (10ml) in add carbonyl diurethane Imidazoles (162mg, 1mmol), is warming up to backflow, and reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue is through rapid column chromatography Separate (dichloromethane: methanol=5:1) and obtain faint yellow solid compound f:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (72mg, yield 74%).ms(esi)m/z: [m+h]+=394.
Step 7:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- base) -1 The preparation of hydrogen-h-benzimidazole-4-carboxamide
Will be dissolved with compound f(72mg, 0.18mmol) tetrahydrofuran solution (5ml) in add ammonia (5ml), be warming up to 70 DEG C, tube sealing reaction cooled down after 8 hours, removal of solvent under reduced pressure, residue through rapid column chromatography separate (dichloromethane: methanol= 10:1) obtain compound as white solid (1): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) Piperazine -1- base) -1 hydrogen-h-benzimidazole-4-carboxamide (7mg, yield 11%).lc-ms(esi):m/z379(m+1)+.1h nmr (300mhz,dmso-d6):δ11.25(br,1h),10.94(br,1h),7.93(s,1h),7.76-7.74(m,1h),7.65- 7.57(m,3h),7.40-7.37(m,1h),7.12-7.06(m,1h),6.64-6.60(m,1h),3.78(br,8h).
Embodiment 2
Compound (2): 2- (4- (8- oxo -8,9- dihydro -7 hydrogen-purine -6- base) piperazine -1- base) -1 hydrogen-benzo miaow The preparation of azoles -4- Methanamide, concrete reaction equation is as follows:
Step 1:2- (4- (6- amino -5- nitro-pyrimidine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- formic acid first The preparation of ester
The method being similar to using embodiment 1 prepare compound d, by compound c and 6- chloro- 5- nitro -4- aminopyrimidine Occur nucleophilic substitution that compound g:2- (4- (6- amino -5- nitro-pyrimidine -4- base) piperazine -1- base) -1- hydrogen-benzo is obtained Imidazoles -4- methyl formate (150mg, yield 93%).ms(esi)m/z:[m+h]+=399.
Step 2:2- (4- (5,6- di-amino-pyrimidine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate Preparation
, there is catalytic hydrogenation system by compound g with palladium carbon in the method being similar to using embodiment 1 prepare compound e Obtain compound h:2- (4- (5,6- di-amino-pyrimidine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate (126mg, yield 69%).ms(esi)m/z:[m+h]+=354.
Step 3:2- (4- (8- oxo -8,9- dihydro -7 hydrogen-purine -6- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- The preparation of methyl formate
, there is ring-closure reaction by compound h with carbonyl dimidazoles in the method being similar to using embodiment 1 prepare compound f Prepared compound i:2- (4- (8- oxo -8,9- dihydro -7 hydrogen-purine -6- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- first Sour methyl ester (72mg, yield 63%).ms(esi)m/z:[m+h]+=395.
Step 4:2- (4- (8- oxo -8,9- dihydro -7 hydrogen-purine -6- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- The preparation of Methanamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound i and ammonia generation ester Prepared compound (2): 2- (4- (8- oxo -8,9- dihydro -7 hydrogen-purine -6- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- Methanamide (44mg, yield 33%).lc-ms(esi):m/z380(m+1)+.1h nmr(300mhz,dmso-d6):δ11.85 (br,1h),11.54(br,1h),10.90(br,1h),9.14(br,1h),8.13(s,1h),7.61(d,1h,j=7.8hz), 7.54(br,1h),7.33(d,1h,j=7.8hz),6.99(t,1h,j=7.8hz),3.68(br,8h).
Embodiment 3
Compound (3): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridin-4-yl) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
Step 1:2- (4- (6- amino -5- nitropyridine -4- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- formic acid first The preparation of ester
The method being similar to using embodiment 1 prepare compound d, by compound c and 2- chloro- 3- nitro -4- aminopyrimidine Occur nucleophilic substitution that compound j:2- (4- (6- amino -5- nitropyridine -4- base) piperazine -1- base) -1- hydrogen-benzo is obtained Imidazoles -4- methyl formate (150mg, yield 92%).ms(esi)m/z:[m+h]+=398.
Step 2:2- (4- (3,4- diamino-pyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate Preparation
, there is catalytic hydrogenation system by compound j with palladium carbon in the method being similar to using embodiment 1 prepare compound e Compound k:2- (4- (3,4- diamino-pyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate (94mg, Yield 100%).ms(esi)m/z:[m+h]+=353.
Step 3:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridin-4-yl) piperazine -1- base) -1 The preparation of hydrogen-benzimidazole -4- methyl formate
, there is ring-closure reaction by compound k with carbonyl dimidazoles in the method being similar to using embodiment 1 prepare compound f Prepared compound l:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridin-4-yl) piperazine -1- base) -1 hydrogen - Benzimidazole -4- methyl formate (82mg, yield 83%).ms(esi)m/z:[m+h]+=394.
Step 4:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridin-4-yl) piperazine -1- base) -1 The preparation of hydrogen-h-benzimidazole-4-carboxamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound l and ammonia generation ester Prepared compound (3): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridin-4-yl) piperazine -1- base) -1 Hydrogen-h-benzimidazole-4-carboxamide (72mg, yield 53%).lc-ms(esi):m/z379(m+1)+.1h nmr(300mhz, dmso-d6):δ11.86(br,1h),11.04(br,1h),10.93(br,1h),9.17(br,1h),7.79(d,1h,j= 5.7hz),7.65-7.52(m,2h),7.32(d,1h,j=8.7hz),7.01-6.98(m,1h),6.66(d,1h,j=5.7hz), 3.72(br,8h).
Embodiment 4
Compound (4): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
Step 1:2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- formic acid first The preparation of ester
The method being similar to using embodiment 1 prepare compound d, by compound c and 6- chloro- 3- nitro -2- aminopyrimidine Occur nucleophilic substitution that compound m:2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -1- hydrogen-benzo is obtained Imidazoles -4- methyl formate (690mg, yield 91%).ms(esi)m/z:[m+h]+=398.
Step 2:2- (4- (5,6- diamino-pyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate Preparation
, there is catalytic hydrogenation system by compound m with palladium carbon in the method being similar to using embodiment 1 prepare compound e Obtain compound n:2- (4- (5,6- diamino-pyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate (290mg, yield 69%).ms(esi)m/z:[m+h]+=353.
Step 3:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 The preparation of hydrogen-benzimidazole -4- methyl formate
, there is ring-closure reaction by compound n with carbonyl dimidazoles in the method being similar to using embodiment 1 prepare compound f Prepared compound o:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen - Benzimidazole -4- methyl formate (73mg, yield 69%).ms(esi)m/z:[m+h]+=394.
Step 4:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 The preparation of hydrogen-h-benzimidazole-4-carboxamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound o and ammonia generation ester Prepared compound (4): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 Hydrogen-h-benzimidazole-4-carboxamide (15mg, yield 22%).lc-ms(esi):m/z379(m+1)+.1h nmr(300mhz, dmso-d6):δ11.85(br,1h),10.99(br,1h),10.40(br,1h),9.15(br,1h),7.60(d,1h,j= 7.5hz),7.51(br,1h),7.33(d,1h,j=7.5hz),7.12(d,1h,j=8.1hz),6.99(t,1h,j=7.5hz), 6.45(d,1h,j=8.1hz),3.67(br,4h),3.48(br,4h).
Embodiment 5
Compound (5): 2- (4- (1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- The preparation of Methanamide, concrete reaction equation is as follows:
Step 1:2- (4- (1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- formic acid The preparation of methyl ester
At one dissolved with compound n(200mg, 0.55mmol) trimethyl orthoformate solution (645mg) in add to first Benzenesulfonic acid (10mg, 0.05mmol), is warming up to backflow, and reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue is through quick post Chromatography (dichloromethane: methanol=5:1) obtain faint yellow solid compound p:2- (4- (1 hydrogen-imidazo [4,5-b] pyridine- 5- yl) piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (97mg, yield 48%).ms(esi)m/z:[m+h]+=378.
Step 2:2- (4- (1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- formyl The preparation of amine
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound p and ammonia generation ester Prepared compound (5): 2- (4- (1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- formyl Amine (32mg, yield 34%).lc-ms(esi):m/z363(m+1)+.1h nmr(300mhz,dmso-d6):δ12.54(br, 1h),9.17(br,1h),8.04(br,1h),7.85-7.60(m,2h),7.61-7.59(m,1h),7.52(br,1h),7.39- 7.32(m,1h),7.01-6.96(m,1h),6.89-6.85(m,1h),3.70-3.68(m,1h).
Embodiment 6
Compound (6): 2- (4- (2- methyl isophthalic acid hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo The preparation of imidazoles -4- Methanamide, concrete reaction equation is as follows:
Step 1:2- (4- (2- methyl isophthalic acid hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo miaow The preparation of azoles -4- methyl formate
At one dissolved with compound n(200mg, 0.55mmol) acetic acid solution (3ml) in add acetic anhydride (61mg, 0.6mmol), it is warming up to backflow, reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue separates (dichloro through rapid column chromatography Methane: methanol=5:1) obtain faint yellow solid compound q:2- (4- (2- methyl isophthalic acid hydrogen-imidazo [4,5-b] pyridine -5- base) Piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (95mg, yield 32%).ms(esi)m/z:[m+h]+=392.
Step 2:2- (4- (2- methyl isophthalic acid hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo miaow The preparation of azoles -4- Methanamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound q and ammonia generation ester Prepared compound (6): 2- (4- (2- methyl isophthalic acid hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo miaow Azoles -4- Methanamide (20mg, yield 23%).lc-ms(esi):m/z377(m+1)+.1h nmr(300mhz,dmso-d6):δ 11.87(br,1h),9.13(br,1h),7.70(d,1h,j=9.3hz),7.60(d,1h,j=7.5hz),7.52(br,1h), 7.33(d,1h,j=7.5hz),6.99(t,1h,j=7.5hz),6.79(d,1h,j=9.3hz),3.69(br,4h),3.62(br, 4h),2.42(s,3h).Embodiment 7
Compound (7): 2- (4- (1 hydrogen-[1,2,3] triazol [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo The preparation of imidazoles -4- Methanamide, concrete reaction equation is as follows:
Step 1:2- (4- (1 hydrogen-[1,2,3] triazol [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo miaow The preparation of azoles -4- methyl formate
At one dissolved with compound n(250mg, 0.68mmol) acetic acid solution (3ml) in add sodium nitrite (47mg, 0.68mmol), it is warming up to backflow, reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue separates (dichloro through rapid column chromatography Methane: methanol=5:1) obtain faint yellow solid compound r:2- (4- (1 hydrogen-[1,2,3] triazol [4,5-b] pyridine -5- base) Piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (230mg, yield 90%).ms(esi)m/z:[m+h]+=379.
Step 2:2- (4- (1 hydrogen-[1,2,3] triazol [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo miaow The preparation of azoles -4- Methanamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound r and ammonia generation ester Prepared compound (7): 2- (4- (1 hydrogen-[1,2,3] triazol [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo miaow Azoles -4- Methanamide (10mg, yield 5%).lc-ms(esi):m/z364(m+1)+.1h nmr(300mhz,dmso-d6):δ 11.86(br,1h),9.14(br,1h),8.17(d,1h,j=9.0hz),7.62-7.53(m,3h),7.33(d,1h,j= 7.8hz),7.13(d,1h,j=9.0hz),6.99(t,1h,j=7.8hz),3.83(br,4h),3.71(br,4h).
Embodiment 8
Compound (8): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
Step 1:2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- formic acid first The preparation of ester
The method being similar to using embodiment 1 prepare compound d, by compound c and 2- chloro- 5- nitro -4- aminopyrimidine Occur nucleophilic substitution that compound s:2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -1- hydrogen-benzo is obtained Imidazoles -4- methyl formate (140mg, yield 90%).ms(esi)m/z:[m+h]+=398.
Step 2:2- (4- (4,5- diamino-pyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate Preparation
, there is catalytic hydrogenation system by compound s with palladium carbon in the method being similar to using embodiment 1 prepare compound e Obtain compound t:2- (4- (4,5- diamino-pyridine -2- base) piperazine -1- base) -1- hydrogen-benzimidazole -4- methyl formate (100mg, yield 100%).ms(esi)m/z:[m+h]+=353.
Step 3:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- base) -1 The preparation of hydrogen-benzimidazole -4- methyl formate
, there is ring-closure reaction by compound t with carbonyl dimidazoles in the method being similar to using embodiment 1 prepare compound f Prepared compound u:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- base) -1 hydrogen - Benzimidazole -4- methyl formate (68mg, yield 62%).ms(esi)m/z:[m+h]+=394.
Step 4:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- base) -1 The preparation of hydrogen-h-benzimidazole-4-carboxamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound u and ammonia generation ester Prepared compound (8): 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- base) -1 Hydrogen-h-benzimidazole-4-carboxamide (32mg, yield 33%).lc-ms(esi):m/z379(m+1)+.1h nmr(300mhz, dmso-d6):δ11.85(br,1h),10.88(br,1h),10.48(br,1h),9.15(br,1h),7.73(s,1h),7.61- 7.59(m,1h),7.51(br,1h),7.33-7.30(m,1h),7.03-6.95(m,1h),6.46(s,1h),3.67-3.65 (m,4h),3.52-3.49(m,4h).
Embodiment 9
Compound (9): 2- (4- (2- trifluoromethyl -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen - The preparation of h-benzimidazole-4-carboxamide, concrete reaction equation is as follows:
Step 1:2- (4- (2- Trifluoromethyl-1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo The preparation of imidazoles -4- methyl formate
At one dissolved with compound n(180mg, 0.49mmol) trifluoroacetic acid solution (5ml) in add trifluoroacetic anhydride (103mg, 0.49mmol), is warming up to backflow, and reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue divides through rapid column chromatography Obtain faint yellow solid compound v:2- (4- (2- Trifluoromethyl-1 hydrogen-imidazo [4,5-b] from (dichloromethane: methanol=5:1) Pyridine -5- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (80mg, yield 37%).ms(esi)m/z:[m+h]+= 446.
Step 2:2- (4- (2- Trifluoromethyl-1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo The preparation of imidazoles -4- Methanamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound v and ammonia generation ester Prepared compound (9): 2- (4- (2- Trifluoromethyl-1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzo Imidazoles -4- Methanamide (18mg, yield 24%).lc-ms(esi):m/z431(m+1)+.1h nmr(300mhz,dmso-d6):δ 13.99(br,1h),11.87(br,1h),9.16(br,1h),7.99-7.96(m,1h),7.63-7.60(m,1h),7.55 (br,1h),7.33(d,1h,j=9.0hz),7.09-6.96(m,2h),3.74-3.72(m,8h).
Embodiment 10
Compound (10): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine Piperazine -1- base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
Fluoro- 3- nitro -2- (2,2,2- trifluoroacetamide) the benzoic preparation of step 1:5-
Under ice bath, fluoro- for 2- trifluoroacetamide -5- benzoic acid (2.5g, 10mmol) is slowly added fuming nitric aicd (14ml), in, reaction is poured in frozen water after continuing stirring under ice bath 1 hour, is filtrated to get compound as white solid w:5- fluoro- 3- nitro -2- (2,2,2- trifluoroacetamide) benzoic acid (1.9g, yield 65%).ms(esi)m/z:[m-h]-=295.
The preparation of step 2:2- amido -5- fluoro- 3- nitrobenzoic acid
By 10% sodium hydrate aqueous solution (20ml) add dissolved with compound w(1.18g, 4mmol) ethanol solution (20ml) In, reaction is warming up to 80 DEG C and stirs 3 hours.Ethanol is removed under reduced pressure, residue adjusts ph to 4 with hydrochloric acid, filters, obtain yellow solid The fluoro- 3- nitrobenzoic acid of compound x:2- amido -5- (0.72g, yield 90%) ms (esi) m/z:[m-h] -=199.
The preparation of step 3:2- amido -5- fluoro- 3- nitrobenzene methyl
Under ice bath, thionyl chloride (2.38g) is slowly added dropwise into dissolved with compound x(0.8g, 4mmol) methanol solution (20ml) in, it is heated to flowing back, reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue separates (oil through rapid column chromatography Ether: ethyl acetate=5:1) obtain yellow solid compound y:2- amido -5- fluoro- 3- nitrobenzene methyl (0.5g, yield 58%).ms(esi)m/z:[m+h]+=215.
The preparation of step 4:2,3- bis- amido -5- fluorophenyl carbamate
The method being similar to using embodiment 1 prepare compound e, by the prepared compound z of compound y generation catalytic hydrogenation: 2,3- bis- amido -5- fluorophenyl carbamates (812mg, yield 46%).ms(esi)m/z:[m+h]+=185.
The preparation of step 5:6- fluoro- 2- oxo -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate
The method being similar to using embodiment 1 prepare compound a, is obtained with carbonyl dimidazoles (cdi) cyclization by compound z Compound a ': 6- fluoro- 2- oxo -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate (711mg, yield 37%).ms(esi) m/z:[m+h]+=211.
The preparation of fluoro- 1 hydrogen of the chloro- 6- of step 6:2--benzimidazole -4- methyl formate
The method being similar to using embodiment 1 prepare compound b, by compound a ' there is chlorination system with phosphorus oxychloride Obtain fluoro- 1 hydrogen of the chloro- 6- of compound b ': 2--benzimidazole -4- methyl formate (681mg, yield 94%).ms(esi)m/z:[m+h]+ =229.
The preparation of the fluoro- 2- of step 7:6- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate
, there is nucleophilic substitution system by compound b ' with piperazine in the method being similar to using embodiment 1 prepare compound c Obtain the fluoro- 2- of compound c ': 6- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (430mg, yield 65%).ms(esi) m/z:[m+h]+=279.
Step 8:2- (4- (2- amino -3- nitropyridine -4- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- first The preparation of sour methyl ester
The method being similar to using embodiment 1 prepare compound d, by compound c ' and chloro- 3 nitros of 4--PA Occur nucleophilic substitution that compound d ': 2- (4- (2- amino -3- nitropyridine -4- base) piperazine -1- base) -6- fluoro- 1- is obtained Hydrogen-benzimidazole -4- methyl formate (310mg, yield 95%).ms(esi)m/z:[m+h]+=416.
Step 9:2- (4- (2,3 diamino pyridine -4- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- formic acid The preparation of methyl ester
The method being similar to using embodiment 1 prepare compound e, occurs catalytic hydrogenation that compound is obtained by compound d ' E ': 2- (4- (2,3- diamino-pyridine -4- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- methyl formate (120mg, Yield 100%).ms(esi)m/z:[m+h]+=386.
The fluoro- 2- of step 10:6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- Base) -1 hydrogen-benzimidazole -4- methyl formate preparation
The method being similar to using embodiment 1 prepare compound f, by compound e ' and carbonyl dimidazoles (cdi) cyclization system Obtain the fluoro- 2- of compound f ': 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- base) -1 Hydrogen-benzimidazole -4- methyl formate (76mg, yield 64%).ms(esi)m/z:[m+h]+=412.
The fluoro- 2- of step 11:6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound f ' and ammonia generation ester Prepared compound (10): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridin-7-yl) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide (28mg, yield 34%).lc-ms(esi):m/z397(m+1)+.1h nmr (300mhz,dmso-d6):δ12.07(br,1h),11.19(br,1h),10.92(br,1h),9.11(br,1h),7.73(d, 1h,j=5.4hz),7.33-7.29(m,1h),7.19-7.16(m,1h),7.02(br,1h),6.58(d,1h,j=5.4hz), 3.72(br,4h),3.21(br,4h).
Embodiment 11
Compound (11): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine Piperazine -1- base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
Step 1:2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- first The preparation of sour methyl ester
The method being similar to using embodiment 1 prepare compound d, by compound c ' and 2- chloro- 5- nitro -4- aminopyrimidine Occur nucleophilic substitution that compound g ': 2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -6- fluoro- 1- is obtained Hydrogen-benzimidazole -4- methyl formate (100mg, yield 73%).ms(esi)m/z:[m+h]+=416.
Step 2:2- (4- (4,5- diamino-pyridine -2- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- formic acid The preparation of methyl ester
, there is catalytic hydrogenation system by compound g ' with palladium carbon in the method being similar to using embodiment 1 prepare compound e Obtain compound h ': 2- (4- (4,5- diamino-pyridine -2- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- methyl formate (40mg, yield 100%).ms(esi)m/z:[m+h]+=371.
Step 3:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- base) -6- The preparation of fluoro- 1 hydrogen-benzimidazole -4- methyl formate
, there is ring-closure reaction by compound h ' with carbonyl dimidazoles in the method being similar to using embodiment 1 prepare compound f Prepared compound i ': 2- (4- (4,5- diamino-pyridine -2- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- formic acid first Ester (48mg, yield 72%).ms(esi)m/z:[m+h]+=412.
The fluoro- 2- of step 4:6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound i ' and ammonia generation ester Prepared compound (11): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-c] pyridine -6- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide (9mg, yield 32%).lc-ms(esi):m/z397(m+1)+.1h nmr(300mhz, dmso-d6):δ12.03(br,1h),10.89(br,1h),10.50(br,1h),9.11(br,1h),7.72(br,2h), 7.32-7.28(m,1h),7.18-7.14(m,1h),6.45(s,1h),3.65(br,4h),3.50(br,4h).
Embodiment 12
Compound (12): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine Piperazine -1- base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
Step 1:2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- first The preparation of sour methyl ester
The method being similar to using embodiment 1 prepare compound d, by compound c ' and 6- chloro- 3- nitro -2- aminopyrimidine Occur nucleophilic substitution that compound j ': 2- (4- (6- amino -5- nitropyridine -2- base) piperazine -1- base) -6- fluoro- 1- is obtained Hydrogen-benzimidazole -4- methyl formate (110mg, yield 91%).ms(esi)m/z:[m+h]+=416.
Step 2:2- (4- (5,6- diamino-pyridine -2- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- formic acid The preparation of methyl ester
, there is catalytic hydrogenation system by compound j ' with palladium carbon in the method being similar to using embodiment 1 prepare compound e Obtain compound k ': 2- (4- (5,6- diamino-pyridine -2- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- methyl formate (90mg, yield 66%).ms(esi)m/z:[m+h]+=371.
Step 3:2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -6- The preparation of fluoro- 1 hydrogen-benzimidazole -4- methyl formate
, there is ring-closure reaction by compound k ' with carbonyl dimidazoles in the method being similar to using embodiment 1 prepare compound f Prepared compound l ': 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -6- Fluoro- 1 hydrogen-benzimidazole -4- methyl formate (73mg, yield 78%).ms(esi)m/z:[m+h]+=412.
The fluoro- 2- of step 4:6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound l ' and ammonia generation ester Prepared compound (4): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide (10mg, yield 32%).lc-ms(esi):m/z397(m+1)+.1h nmr (300mhz,dmso-d6):δ12.05(br,1h),11.00(br,1h),10.42(br,1h),9.10(br,1h),7.72(br, 1h),7.32(br,1h),7.18-7.10(m,2h),6.43(d,1h,j=8.7hz),3.66(br,4h),3.48(br,4h).
Embodiment 13
Compound (13): the fluoro- 2- of 6- (4- (2- trifluoromethyl -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
The fluoro- 2- of step 1:6- (4- (2- trifluoromethyl -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 The preparation of hydrogen-benzimidazole -4- methyl formate
Trifluoroacetic anhydride is added in a trifluoroacetic acid solution (5ml) dissolved with compound k ' (180mg, 0.49mmol) (103mg, 0.49mmol), is warming up to backflow, and reaction cooled down after 8 hours, removal of solvent under reduced pressure, and residue divides through rapid column chromatography Obtain the fluoro- 2- of faint yellow solid compound m ': 6- (4- (2- trifluoromethyl -3 hydrogen-imidazo from (dichloromethane: methanol=5:1) [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate (65mg, yield 31%).ms(esi)m/z: [m+h]+=464.
The fluoro- 2- of step 2:6- (4- (2- trifluoromethyl -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 The preparation of hydrogen-h-benzimidazole-4-carboxamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound m ' and ammonia generation ester Prepared compound (13): the fluoro- 2- of 6- (4- (2- trifluoromethyl -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 Hydrogen-h-benzimidazole-4-carboxamide (10mg, yield 16%).lc-ms(esi):m/z449(m+1)+.1h nmr(300mhz, dmso-d6):δ14.07(br,1h),12.56(br,1h),9.12(br,1h),7.98(br,1h),7.73(br,1h),7.58- 7.24(m,3h),3.73(br,8h).
Embodiment 14
Compound (14): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -6- base) piperazine Piperazine -1- base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation, concrete reaction equation is as follows:
Step 1:2- (4- (5- amino -6- nitropyridine -3- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- first The preparation of sour methyl ester
The method being similar to using embodiment 1 prepare compound d, by compound c ' and 5- bromo- 2- nitro -3- aminopyrimidine Occur nucleophilic substitution that compound n ': 2- (4- (5- amino -6- nitropyridine -3- base) piperazine -1- base) -6- fluoro- 1- is obtained Hydrogen-benzimidazole -4- methyl formate (210mg, yield 73%).ms(esi)m/z:[m+h]+=416.
Step 2:2- (4- (5,6- diamino-pyridine -3- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- formic acid The preparation of methyl ester
, there is catalytic hydrogenation system by compound n ' with palladium carbon in the method being similar to using embodiment 1 prepare compound e Obtain compound o ': 2- (4- (5,6- diamino-pyridine -3- base) piperazine -1- base) -6- fluoro- 1- hydrogen-benzimidazole -4- methyl formate (91mg, yield 68%).ms(esi)m/z:[m+h]+=371.
The fluoro- 2- of step 3:6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -6- base) piperazine -1- Base) -1 hydrogen-benzimidazole -4- methyl formate preparation
, there is ring-closure reaction by compound o ' with carbonyl dimidazoles in the method being similar to using embodiment 1 prepare compound f Fluoro- 2- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -6- base) piperazine -1- of prepared compound p ': 6- Base) -1 hydrogen-benzimidazole -4- methyl formate (63mg, yield 73%).ms(esi)m/z:[m+h]+=412.
The fluoro- 2- of step 4:6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -6- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide preparation
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound p ' and ammonia generation ester Prepared compound (14): the fluoro- 2- of 6- (4- (2- oxo -2,3- dihydro -1 hydrogen-imidazo [4,5-b] pyridine -6- base) piperazine -1- Base) -1 hydrogen-h-benzimidazole-4-carboxamide (12mg, yield 31%).lc-ms(esi):m/z449(m+1)+.1h nmr (300mhz,dmso-d6):δ12.04(br,1h),10.95(br,1h),10.48(br,1h),9.10(br,1h),7.71(br, 1h),7.31-7.28(m,2h),7.15(s,1h),6.44(s,1h),3.67(br,4h),3.49(br,4h).
Embodiment 15
Compound (15): the fluoro- 2- of 6- (4- (2- methoxyl group -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) - The preparation of 1 hydrogen-h-benzimidazole-4-carboxamide, concrete reaction equation is as follows:
Step 1:2- (4- (chloro- 1 hydrogen of 2--imidazo [4,5-b] pyridine -5- base) piperazine -1- base) fluoro- 1 hydrogen of -6--benzo The preparation of imidazoles -4- methyl formate
, there is chlorination system by compound l ' with phosphorus oxychloride in the method being similar to using embodiment 1 prepare compound b Obtain compound q ': 2- (4- (chloro- 1 hydrogen of 2--imidazo [4,5-b] pyridine -5- base) piperazine -1- base) fluoro- 1 hydrogen of -6--benzo miaow Azoles -4- methyl formate (381mg, yield 74%).ms(esi)m/z:[m+h]+=430.
The fluoro- 2- of step 2:6- (4- (2- methoxyl group -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen - The preparation of h-benzimidazole-4-carboxamide
, there is nucleophilic substitution by compound q ' with Feldalat NM in the method being similar to using embodiment 1 prepare compound d Prepared compound r ': 6- fluoro- 2- (4- (2- methoxyl group -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen-benzene And imidazoles -4- Methanamide (132mg, yield 51%).ms(esi)m/z:[m+h]+=426.
The fluoro- 2- of step 3:6- (4- (2- methoxyl group -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen - The preparation of h-benzimidazole-4-carboxamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound r ' and ammonia generation ester Prepared compound (15): the fluoro- 2- of 6- (4- (2- methoxyl group -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -1 hydrogen - H-benzimidazole-4-carboxamide (13mg, yield 33%).lc-ms(esi):m/z411(m+1)+.1h nmr(300mhz,dmso- d6):δ12.03(br,1h),9.09(br,1h),7.96(br,1h),7.72-7.61(m,2h),7.32(d,1h,j=7.5hz), 6.80(d,1h,j=7.5hz),3.82(s,3h),3.67(br,4h),3.61(br,4h).
Embodiment 16
Compound (16): 2- (4- (2- dimethylamino -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -6- The preparation of fluoro- 1 hydrogen-h-benzimidazole-4-carboxamide, concrete reaction equation is as follows:
Step 1:2- (4- (2- dimethylamino -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -6- fluoro- 1 The preparation of hydrogen-benzimidazole -4- methyl formate
, there is nucleophilic substitution by compound q ' with dimethylamine in the method being similar to using embodiment 1 prepare compound d Prepared compound s ': 2- (4- (2- dimethylamino -1 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) fluoro- 1 hydrogen of -6- - Benzimidazole -4- methyl formate (72mg, yield 33%).ms(esi)m/z:[m+h]+=439.
Step 2:2- (4- (2- dimethylamino -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -6- fluoro- 1 The preparation of hydrogen-h-benzimidazole-4-carboxamide
The method being similar to using embodiment 1 prepare compound (1), by the ammonolysis reaction of compound s ' and ammonia generation ester Prepared compound (16): 2- (4- (2- dimethylamino -3 hydrogen-imidazo [4,5-b] pyridine -5- base) piperazine -1- base) -6- fluoro- 1 Hydrogen-h-benzimidazole-4-carboxamide (15mg, yield 31%).lc-ms(esi):m/z424(m+1)+.1h nmr(300mhz, dmso-d6):δ12.05(br,1h),9.06(br,1h),7.95(br,1h),7.73-7.63(m,2h),7.31(d,1h,j= 7.2hz),6.82(d,1h,j=7.2hz),3.69(br,4h),3.60(br,4h),3.14(s,6h).
Biological assessment
Experimental principle:
The poly adp of nucleoprotein is ribosylating be the translation occurring when dna damages response after.Parp, full name is poly- adenosine Diphosphonic acid ribose polymerase, in the presence of having nad, catalysis poly (adp- ribose) is connected on the nucleoprotein closing on, thus drawing Send out the dna repair mechanism via base excision repair path.The ht universal that trevigen company produces Chemiluminescent parp assay kit can measure this adp- ribose by biotin labeling and histone In conjunction with level.
Reagent and consumptive material
1.ht universal chemiluminescent parp assay kit with histone-coated Strip wells, U.S. trevigen, article No.: 4676-096-k
2. plate reader, U.S. perkin elmer, envision multilabel plate reader
Solution and buffer
1. washing liquid contains the pbs solution of 0.1%triton x-100
The dilution of 20x parp buffer is obtained 1x buffer for 20 times by 2.20x parp buffer deionized water, should Buffer is used to dilution restructuring parp enzyme, parp cocktail and tested compound.
3.10x parp cocktail prepares 1x parp cocktail:10x parp in accordance with the following methods Cocktail2.5 μ l/well, 10x activate dna2.5 μ l/well, 1x parp buffer 20 μ l/well.
4.parp enzyme only before use, carefully dilutes recombinase with 1x parp buffer, the enzymatic solution having diluted To use as early as possible, unspent will discard.
5.strep-hrp only before use, obtains 1x solution with 1x strep diluted strep-hrp500 times.
6. chemical luminous substrate only before use, the peroxyglow a of same volume and b solution mix homogeneously is obtained The substrate of horseradish peroxidase.
Experimental technique
Compound is prepared
1. with dmso, each for 10mm test compound mother solution is diluted to 10um, 1um.
2., only before experiment starts, the gradient concentration solution 1x parp of each compound being dissolved in dmso delays Rush liquid and dilute 20 times, obtain the compound solution of 5x, you can for being detected, positive control (positive) and negative control (negative) hole is 1x parp buffer (dmso content 5%)
Operating procedure
1. every hole adds 50 μ l1x parp buffer moistening histones, in incubated at room orifice plate 30 minutes, then by hole 1x parp buffer suction out, and on napkin, residual liquid is patted dry only.
2., according to compound (1)～(16), the 5x having diluted compound solution is added in corresponding hole, every hole 10 μ l, Positive control (positive) and negative control (negative) hole are 1x parp buffer (dmso content 5%)
3. with 1x parp buffer, parp enzyme is diluted to every 15 μ l solution and contains 0.5unit, then except negative right Add 15 μ l enzymatic solution according to other holes beyond hole, negative control hole only adds 1x parp buffer, 10 points of incubated at room orifice plate Clock.
4. the 1x parp cocktail continuously adding 25 μ l is in each hole.
5.27 DEG C are incubated orifice plate 60 minutes.
6., after incubation terminates, the reactant liquor in hole is suctioned out, and on napkin, residual liquid is patted dry only.Then with containing The pbs solution of 0.1%triton x-100 rinses orifice plate 4 times, and every hole is with 200 μ l every time, and pats dry residual liquid on napkin Only.
7., next, adding the 1x strep-hrp solution having diluted in every hole, then it is incubated 60 points of orifice plate at 27 DEG C Clock.
8., after incubation terminates, the reactant liquor in hole is suctioned out, and on napkin, residual liquid is patted dry only.Then with containing The pbs solution of 0.1%triton x-100 rinses orifice plate 4 times, and every hole is with 200 μ l every time, and pats dry residual liquid on napkin Only.
9., after washing hardened bundle, by the peroxyglow a of same volume and b solution mix homogeneously, every hole adds 100 μ l, stands Put into plate reader record chemiluminescence signal.
Data processing
Reading in every hole needs to be converted into suppression ratio.The suppression ratio of compound can be calculated using following equation Go out:
Note: Positive control wells reading is positive hole reading, meaning is enzyme 100% activity；Negative control hole reading is Negative hole reading, meaning is enzyme 0%；Active x is the reading of each each concentration of sample.
The inhibitory activity to parp-1 enzyme for table 1 compound
Compound number The suppression ratio (%) of 100nm concentration The suppression ratio (%) of 30nm concentration ic50Value
(1) 78 65 24nm
(2) 64 38 64nm
(3) 55 31 94nm
(4) 83 71 8nm
(5) 72 57 26nm
(6) 68 50 37nm
(7) 77 56 28nm
(8) 76 63 21nm
(9) 47 33 195nm
(10) 85 74 13nm
(11) 82 71 14nm
(12) 86 76 9nm
(13) 50 30 93nm
(14) 81 73 17nm
(15) 63 47 62nm
(16) 58 36 84nm
The activity data listed in table 1 fully shows, the compound of the present invention is all the inhibitor of parp-1, wherein implements In example, compound (1), (2), (3), (4), (5), (6), (7), (8), (10), (11), (12), (13), (14), (15), (16) Ic50 value be not more than 100nm, compound (4), the ic50 value of (13) no more than 10nm.

Claims (27)

1. pyrimido glyoxaline compound, it is formula () or the compound shown in formula () or it is pharmaceutically acceptable Salt:
Wherein, in formula () or formula ():
R is hydrogen or halogen；
X, y, z is one of to be nitrogen, remaining be hydrocarbon or x, y, z one of for hydrocarbon, remaining is nitrogen；
M is nitrogen or cr1；
R1 is hydrogen, oxygen, c1-c6 alkyl, methoxyl group, trifluoromethyl and nr2r3；
R2 is hydrogen or c1-c4 alkyl；
R3 is hydrogen or c1-c4 alkyl.
2. pyrimido glyoxaline compound according to claim 1 is it is characterised in that described formula () or formula () Shown compound or its pharmaceutically acceptable salt, wherein:
R is hydrogen or fluorine；
X, y, z is one of to be nitrogen, and remaining is hydrocarbon；
M is nitrogen or cr1；
R1 is hydrogen, oxygen, c1-c4 alkyl, methoxyl group, trifluoromethyl and nr2r3；
R2 is hydrogen or c1-c4 alkyl；
R3 is hydrogen or c1-c4 alkyl.
3. pyrimido glyoxaline compound according to claim 1 or its pharmaceutically acceptable salt are it is characterised in that institute Stating the compound shown in formula () is following compound (1)～(3) and (10):
4. pyrimido glyoxaline compound according to claim 1 or its pharmaceutically acceptable salt are it is characterised in that institute Stating the compound shown in formula () is following compound:
5. pyrimido glyoxaline compound as claimed in claim 1 or its pharmaceutically acceptable salt are it is characterised in that described Pharmaceutically acceptable salt is the hydrochlorate of formula () or formula () compound, sulfate, phosphate, acetate, trifluoro second Hydrochlorate, mesylate, fluoroform sulphonate, tosilate, tartrate, maleate, fumarate, succinate or Malate.
6. the preparation method of the compound shown in formula () described in claim 1, its reaction equation is as follows:
Wherein, r, x, y, z and m are as defined above；Specifically comprise the following steps that
Step 1): substituted 2,3- diamidogen yl benzoic acid methyl ester and carbonyl dimidazoles ring-closure reaction, obtain 2- oxo -2 replacing, 3- dihydro -1 hydrogen-benzimidazole -4- methyl formate ()；
Step 2): step 1) substituted 2- oxo -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate () and three of obtaining Chlorethoxyfos carry out chlorination reaction, obtain chloro- 1 hydrogen of the 2--benzimidazole -4- methyl formate () replacing；
Step 3): in the presence of a base, by step 2) substituted chloro- 1 hydrogen of 2--benzimidazole -4- methyl formate () of obtaining with Piperazine carries out nucleophilic substitution, obtains 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate () replacing；
Step 4): by step 3) substituted 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate () of obtaining with many There is nucleophilic substitution in the pyrimidine compound replacing, obtain intermediate ()；
Step 5): by step 4) there is catalytic hydrogen reduction nitro in the intermediate () that obtains, obtain intermediate ()；
Step 6): by step 5) intermediate () that obtains by with acetic anhydride, trifluoro-acetic anhydride, trimethyl orthoformate or nitrine Change sodium and ring-closure reaction occurs, obtain intermediate ()；
Step 7): by step 6) intermediate () that obtains passes through the aminolysis reaction of ester group in methanolic ammonia solution, generates Compound shown in formula ().
7. the preparation method of the compound shown in formula () described in claim 1, its reaction equation is as follows:
Wherein, r, x, y, z and m are as defined above；Comprise the following steps that
Step (1): substituted 2,3- diamidogen yl benzoic acid methyl ester and carbonyl dimidazoles ring-closure reaction, obtain the 2- oxo that replaces- 2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate ()；
Step (2): substituted 2- oxo -2,3- dihydro -1 hydrogen-benzimidazole -4- methyl formate () that step (1) obtains with Phosphorus oxychloride carries out chlorination reaction, obtains chloro- 1 hydrogen of the 2--benzimidazole -4- methyl formate () replacing；
Step (3): in the presence of a base, substituted chloro- 1 hydrogen of 2--benzimidazole -4- methyl formate () that step (2) is obtained Carry out nucleophilic substitution with piperazine, obtain 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate () replacing；
Step (4): substituted 2- (piperazine -1- base) -1 hydrogen-benzimidazole -4- methyl formate () that step (3) is obtained with There is nucleophilic substitution in polysubstituted pyrimidine compound, obtain intermediate ()；
Step (5): the intermediate () that step (4) is obtained occurs catalytic hydrogen reduction nitro, obtains intermediate ()；
Step (6): the intermediate () that step (5) is obtained by with acetic anhydride, trifluoro-acetic anhydride, trimethyl orthoformate or folded There is ring-closure reaction in sodium nitride, obtain intermediate ()；
Step (7): the intermediate () that step (6) is obtained passes through the aminolysis reaction of ester group in methanolic ammonia solution, raw Become the compound shown in formula ().
8. Pharmaceutical composition, comprises the formula () of therapeutically effective amount or formula () compound or its medicine constituting active component Acceptable salt and pharmaceutically acceptable carrier, diluent on.
9. pharmaceutical composition as claimed in claim 8, its described pharmaceutical composition makes tablet, capsule, aqueouss suspension Agent, Oil suspensions, dispersible powder, granule, lozenge, Emulsion, syrup, ointment, ointment, suppository or injection Agent.
10. formula () described in any one of claim 1 to 5 claim or formula () compound or it is pharmaceutically acceptable Application in the preparation disease medicament that improves because of parp activity suppression for the treatment of for the salt.
11. apply as claimed in claim 10, and the described disease improved because of parp activity suppression is angiopathy, septic Shock, ischemic injuries, neurotoxicity, hemorrhagic shock, inflammatory diseasess or multiple sclerosis.
12. claim 8 or 9 described pharmaceutical compositions are in the disease medicament that preparation treatment improves because of parp activity suppression Application.
13. apply as claimed in claim 12, and the described disease improved because of parp activity suppression is angiopathy, septic Shock, ischemic injuries, neurotoxicity, hemorrhagic shock, inflammatory diseasess or multiple sclerosis.
Formula () described in 14. any one of claim 1 to 5 or formula () compound or its pharmaceutically acceptable salt are in system It is ready for use on the application in the ancillary drug of oncotherapy.
15. claim 8 or 9 described pharmaceutical compositions are used for the application in the ancillary drug of oncotherapy in preparation.
Formula () described in 16. claim 1 to 5 any one claim or formula () compound are strong for tumor in preparation Application in the medicine of chemoradiotherapy.
17. claim 8 or 9 described pharmaceutical compositions strengthen the application in the medicine of radiotherapy in preparation for tumor.
Formula () described in 18. claim 1 to 5 any one claim or formula () compound or it is pharmaceutically acceptable Salt preparation for chemotherapy of tumors medicine in application.
19. claim 8 or 9 described pharmaceutical compositions are used for the application in the medicine of chemotherapy of tumors in preparation.
Formula () described in 20. claim 1 to 5 any one claim or formula () compound or it is pharmaceutically acceptable Salt preparation lack homologous recombination dependent dna double-strand break repair individuation treatment of cancer medicine in application.
21. apply as claimed in claim 20, and wherein said cancer is to pass through homologous recombination repair dna containing one or more Double-strand break the ability cancer of cancerous cell that lowers with respect to normal cell or lose.
22. apply as claimed in claim 20, and wherein said cancer is to have brca-1 or brca-2 defect, mutant phenotype Cancer.
23. apply as claimed in claim 20, and described cancer is breast carcinoma, ovarian cancer, cancer of pancreas or carcinoma of prostate.
24. claim 8 or 9 described pharmaceutical compositions lack what homologous recombination dependent dna double-strand break was repaired in preparation Application in the medicine of individuation treatment of cancer.
25. apply as claimed in claim 24, and described cancer is to pass through the double of homologous recombination repair dna containing one or more The cancer of cancerous cell that the ability of chain interruption lowers with respect to normal cell or loses.
26. apply as claimed in claim 24, and described cancer is the cancer with brca-1 or brca-2 defect, mutant phenotype Disease.
27. apply as claimed in claim 24, and described cancer is breast carcinoma, ovarian cancer, cancer of pancreas or carcinoma of prostate.
CN201310165742.5A 2013-05-07 2013-05-07 Pyrimidoimidazole compound and pharmaceutical composition and preparation method and use thereof Active CN104140426B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201310165742.5A CN104140426B (en) 2013-05-07 2013-05-07 Pyrimidoimidazole compound and pharmaceutical composition and preparation method and use thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201310165742.5A CN104140426B (en) 2013-05-07 2013-05-07 Pyrimidoimidazole compound and pharmaceutical composition and preparation method and use thereof

Publications (2)

Publication Number Publication Date
CN104140426A CN104140426A (en) 2014-11-12
CN104140426B true CN104140426B (en) 2017-02-01

Family

ID=51849776

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201310165742.5A Active CN104140426B (en) 2013-05-07 2013-05-07 Pyrimidoimidazole compound and pharmaceutical composition and preparation method and use thereof

Country Status (1)

Country Link
CN (1) CN104140426B (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106146492A (en) * 2015-04-17 2016-11-23 上海汇伦生命科技有限公司 Heterocycle glyoxaline compound, its pharmaceutical composition and its production and use
CN106146504B (en) * 2015-04-17 2018-09-07 上海汇伦生命科技有限公司 A kind of heterocycle and glyoxaline compound, its pharmaceutical composition and its preparation method and application
WO2017013237A1 (en) 2015-07-23 2017-01-26 Institut Curie Use of a combination of dbait molecule and parp inhibitors to treat cancer
WO2018162439A1 (en) 2017-03-08 2018-09-13 Onxeo New predictive biomarker for the sensitivity to a treatment of cancer with a dbait molecule
US11472788B2 (en) 2017-11-25 2022-10-18 Beigene, Ltd. Benzoimidazoles as selective inhibitors of indoleamine 2,3-dioxygenases
EP3765613A1 (en) 2018-03-13 2021-01-20 Onxeo A dbait molecule against acquired resistance in the treatment of cancer
CN113891880A (en) * 2019-05-22 2022-01-04 百济神州有限公司 Amide substituted imidazo compounds as selective inhibitors of indoleamine 2, 3-dioxygenase
EP3976612A4 (en) * 2019-05-28 2023-04-26 Mankind Pharma Ltd Novel compounds for inhibition of janus kinase 1
WO2021148581A1 (en) 2020-01-22 2021-07-29 Onxeo Novel dbait molecule and its use

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1195985A (en) * 1995-08-02 1998-10-14 钮卡斯尔大学风险投资有限公司 Benzimidazole compounds
CN1353695A (en) * 1999-05-07 2002-06-12 巴斯福股份公司 Heterocyclically substituted benzimidazoles, production and application thereof
WO2003106430A1 (en) * 2002-06-14 2003-12-24 Pfizer Inc. Benzimidazole inhibitors of poly(adp-ribosyl) polymerase
CN101163706A (en) * 2005-04-21 2008-04-16 贝林格尔·英格海姆国际有限公司 Dihydrothienopyrimidines for the treatment of inflammatory diseases
CN101754967A (en) * 2007-07-16 2010-06-23 雅培制药有限公司 Benzoglyoxaline gathers (ADP-ribose) AG14361
WO2012122383A2 (en) * 2011-03-09 2012-09-13 Avila Therapeutics, Inc. Pi3 kinase inhibitors and uses thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1195985A (en) * 1995-08-02 1998-10-14 钮卡斯尔大学风险投资有限公司 Benzimidazole compounds
CN1353695A (en) * 1999-05-07 2002-06-12 巴斯福股份公司 Heterocyclically substituted benzimidazoles, production and application thereof
WO2003106430A1 (en) * 2002-06-14 2003-12-24 Pfizer Inc. Benzimidazole inhibitors of poly(adp-ribosyl) polymerase
CN101163706A (en) * 2005-04-21 2008-04-16 贝林格尔·英格海姆国际有限公司 Dihydrothienopyrimidines for the treatment of inflammatory diseases
CN101754967A (en) * 2007-07-16 2010-06-23 雅培制药有限公司 Benzoglyoxaline gathers (ADP-ribose) AG14361
WO2012122383A2 (en) * 2011-03-09 2012-09-13 Avila Therapeutics, Inc. Pi3 kinase inhibitors and uses thereof

Also Published As

Publication number Publication date
CN104140426A (en) 2014-11-12

Similar Documents

Publication Publication Date Title
CN104140426B (en) Pyrimidoimidazole compound and pharmaceutical composition and preparation method and use thereof
EP2604610B1 (en) Phthalazinone ketone derivative, preparation method thereof, and pharmaceutical use thereof
EP3438094A1 (en) Selective c-kit kinase inhibitor
TWI359142B (en) Quinazolinedione derivatives as parp inhibitors
CN104981468B (en) The piperazine heterocycle compound of benzimidazole 2, its pharmaceutical composition and its production and use
CN103242341A (en) Thieno 2,4-substituted pyrimidine compound, and pharmaceutical composition and application thereof
CN108026095A (en) Noval chemical compound
CN104003979B (en) Benzimidazolyl-2 radicals-piperazine compounds, its pharmaceutical composition and its production and use
ES2927959T3 (en) Heterocyclic imidazole compounds, pharmaceutical compositions thereof, method of preparation thereof and use thereof
CN106146504B (en) A kind of heterocycle and glyoxaline compound, its pharmaceutical composition and its preparation method and application
CN103923088B (en) 2,5-diazabicyclo [2.2.1] heptane class compound and preparation method, its Pharmaceutical composition and in application pharmaceutically
AU2017342887B2 (en) Pharmaceutical salt of antitumor heterocyclic imidazole compound
CN104230897B (en) Benzimidazolyl-2 radicals-piperazine heterocycle compound, its pharmaceutical composition and its production and use
CN104230898B (en) Benzimidazolyl-2 radicals-piperazine heterocycle compound, its pharmaceutical composition and its production and use
EP3878841A1 (en) Indazole kinase inhibitor and use thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
TR01 Transfer of patent right

Effective date of registration: 20190515

Address after: Room 304-12, 665 Zhangjiang Road, Shanghai Pudong New Area Free Trade Pilot Area, 201210

Patentee after: Shanghai Huilun Pharmaceutical Technology Co., Ltd.

Address before: Room 650-10, Building No. 2, 351 Guoshoujing Road, Zhangjiang High-tech Park, Pudong New Area, Shanghai, 201203

Patentee before: Shanghai Huilun Technology Co., Ltd.

TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20190814

Address after: Room 650-10, Building No. 2, 351 Guoshoujing Road, Pudong New Area Free Trade Pilot Area, Shanghai, 201203

Patentee after: Shanghai Huilun Technology Co., Ltd.

Address before: Room 304-12, 665 Zhangjiang Road, Shanghai Pudong New Area Free Trade Pilot Area, 201210

Patentee before: Shanghai Huilun Pharmaceutical Technology Co., Ltd.

TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20191226

Address after: 201108 Room 102, floor 1, building 12, No. 1835, Duhui Road, Minhang District, Shanghai

Patentee after: Shanghai Yidian Medical Science and Technology Development Co., Ltd.

Address before: Room 650-10, Building No. 2, 351 Guoshoujing Road, Pudong New Area Free Trade Pilot Area, Shanghai, 201203

Patentee before: Shanghai Huilun Technology Co., Ltd.

TR01 Transfer of patent right