CN106317069A

CN106317069A - 2-substituted-benzimidazole-4-formamide compound, preparation method, and application thereof

Info

Publication number: CN106317069A
Application number: CN201510363842.8A
Authority: CN
Inventors: 杨春皓; 缪泽鸿; 陈旭星; 宦霞娟; 王玉勤; 陈奕; 丁健
Original assignee: Shanghai Institute of Materia Medica of CAS
Current assignee: Shanghai Institute of Materia Medica of CAS
Priority date: 2015-06-26
Filing date: 2015-06-26
Publication date: 2017-01-11
Also published as: CN107922430B; WO2016206651A1; CN107922430A

Abstract

The invention relates to a 2-substituted-benzimidazole-4-formamide compound, a preparation method, and an application thereof. The structure of the 2-substituted-benzimidazole-4-formamide compound is represented as the general formula (I), wherein R1, R2, Y1, Y2, R3 and X are defined as the claims and the specifications. The invention also discloses a medicine composition comprising the 2-substituted-benzimidazole-4-formamide compound or pharmaceutically acceptable salts thereof. The 2-substituted-benzimidazole-4-formamide compound and the medicine composition can be used for preparing medicines for treating diseases related to Poly ADP-ribose polymerase, e.g., malignant tumor.

Description

2-substituted benzimidazole-4-Carbox amide and preparation method and application

Technical field

The invention belongs to pharmaceutical chemistry and pharmacotherapeutics field.It is specifically related to class 2-(thiophene or thiazole tetrahydropyridine-2- Base)-1H-h-benzimidazole-4-carboxamide compounds or its pharmaceutically acceptable salt, its preparation method and they control in preparation Treat the disease such as application in the medicine of malignant tumor relevant to Poly adenosine diphosphate-ribose polymerase-1 (PARP).

Background technology

Every day about 10 in human body¹³Individual cell by tens thousand of DNA damage, be exposed in rugged environment (such as: Ultraviolet radiation, ionizing radiation etc.), normal cellular metabolism by-product, cell toxicity medicament all can cause DNA damage.This A little damages include base modification, single-strand break (Single strand breaks, SSB), double-strand break (Double strand breaks, DSB), interconnection etc..DNA damage has a strong impact on the duplication of gene and transcribes, and even causes genome distortion and tumor Generation.In order to maintain the complete stability of genome, body system exist multiple approach detection DNA damage and per time every Carve and DNA damage is repaired.Main DNA repair approach include base excision repair (Base-excision repair, BER), Nucleotide Sequence Analysis (Nucleotide-excision repair, NER), mispairing reparation (Mismatch repair, MMR), homologous recombination (Homologous recombination, HR) and non-homologous end joining (Nonhomologous End joining, NHEJ) etc..Wherein BER, NER, MMR are that main DNA single-strand break (SSB) repairs approach, HR and NHEJ is that approach is repaired in main DNA double chain interruption (DSB), and HR is approach of reliably repairing accurately, and NHEJ And unstable, easily make mistakes.Additionally some DNA damage directly can be repaired by corresponding enzyme, such as guanine methyl Can be by O-6-methyl guanine-dnmt rna (MGMT) demethylation reparation.Cancer patient is using cell toxicant After medicine, DNA of tumor cell injury repairing increases, and causes apoptosis to tolerate, and is the important former of drug resistance of tumor generation One of because of.

Poly adenosine diphosphate-ribose polymerase-1 (Poly (ADP)-ribose polymerase, PARP) is that a class is present in majority Cell ribozyme in eukaryotic cell.At present, the most 17 PARP family members are found, all PARP families Albumen has similar catalyst structure domain sequence.It is true that PARP family member is only partially with nicotinoyl amine gland fast Nicotinamide adenine dinucleotide (Nicotinamide adenine dinucleotide, NAD⁺) it is that substrate synthesizes poly-adenosine diphosphate ribose (Poly (adenosine diphosphate-ribose), PAR), and transfer them to receptor protein, regulation and control associated protein Function.In PARP family, only PARP1/2 can be activated by DNA single-strand break and to mediate poly-ADP ribosylating, The reparation of DNA damage is participated in via base excision repair approach.Mankind PARP1 be a molecular weight be 113kDa Polypeptide chain, comprises 3 Functional domains.Be positioned at N-end DNA binding structural domain (DNA binding domain, DBD), containing two zinc fingerses (Zinc finger), identify DNA break.1st zinc refer to identify DNA break and The activation of PARP1 activity, its damage will thoroughly make PARP1 loss of activity, and it is mono-that the 2nd zinc refers to be only involved in DNA The identification of chain interruption.Mid portion certainly modify domain (Automodification domain, AMD), PARP1 passes through This district is combined with poly-ADP ribosyl, occurs self poly-ADP ribosylating, regulates the phase interaction of self and DNA or albumen With.The catalyst structure domain (Catalytic domain) of C-end, is the part of PARP1 strict conservation, catalyzes and synthesizes PAR And by poly-for target protein domain ribosylating for ADP.Mankind PARP2 be a molecular weight be the polypeptide chain of 62kDa. Different from PARP1, the N-end of PARP2 does not contains zinc fingers, and the effectiveness that therefore SSB is identified by PARP2 reduces, Then identify the space lost on DNA damage chain and formed due to nucleotide.The catalyst structure domain of PARP2 Yu PARP1 Basic simlarity, has the homology of 69%, but the nuance of structure still reflects the difference of they functions.In order to right The generation of antitumor drug resistance and reduce necrocytosis and the inflammatory reaction that produces, in early days, PARP1 inhibitor is used as Chemical sensitizer, kills tumor cell with Radiotherapy chemotherapy synergy.

Until 2005, two different seminars are simultaneously great prominent Nature delivered PARP research field one Broken: PARP1 inhibitor specificity suppresses/kill the cell that mastocarcinoma gene BRCA1 and BRCA2 lacks.As it was previously stated, DNA double chain reparation includes HR and NHEJ, but NHEJ likely there will be mistake, causes the instability of genome. HR is mainly by two key gene mediations of BRCA1 and BRCA2, and in the cell of BRCA disappearance, DNA plerosis is double Chain interruption needs, by NHEJ approach, to thus result in and be susceptible to tumor.BRCA1/2 disappearance the most easily shows as breast Adenocarcinoma, ovarian cancer, carcinoma of prostate, cancer of pancreas etc., the tumor of this HR approach inactivation is referred to as " BRCAness ".? In the cell of HR defect, suppression PARP1 will cause the accumulation of DNA single-strand break to increase, and be converted into lethal DSBs. The DSBs of lethal causes the unstability of chromosomal aberration and genome, produces cell death.That is, one gene merit Can disappearance will cause sensitizing cells (such as the disappearance of PARP1 or BRCA1/2), but the disappearance of two gene functions But it is fatal (as PARP1 and BRCA1/2 lacks simultaneously).BRCAness Yu PARP1/2 suppression is common to be killed carefully This phenomenon of born of the same parents is referred to as collaborative lethal (Synthetic lethality).The treatment that collaborative lethal theory is malignant tumor provides New Policy and new approaches, since then, the research of PARP1 inhibitor enters brand-new epoch.

Existing PARP1/2 inhibitor is mainly by simulating the substrate NAD of PARP⁺, competitive inhibition PARP activity. Large quantities of different skeleton PARP inhibitor X-ray eutectic researchs provide sufficiently support for PARP1 inhibitor structure activity relationship. Sum up PARP inhibitor structure activity relationship and mainly have the following aspects: maintain activity and adhesion must at least contain one The Methanamide structure that individual hydrogen is free, it can be fused in twin nuclei or " false dicyclo " system, become hydrogen bond receptor simultaneously And hydrogen-bond donor；For changing bigger hydrophobic group of the volume of activity and physicochemical property etc..At present, existing multiple PARP1/2 inhibitor enters clinical trial, and wherein AZD2281 has gone through to list.But, most of PARP1/2 press down It is low all to there is bioavailability in preparation, the shortcoming of subtype-selective difference.

To sum up, still need and research and develop PARP1/2 inhibitor further in this area.

Summary of the invention

It is an object of the invention to provide a kind of 2-substituted benzimidazole-4-Carbox amide and preparation method thereof and answer With.

A first aspect of the present invention, it is provided that the compound shown in a kind of formula I, R-isomer, S-isomer or its pharmacy Upper acceptable salt:

Wherein,

R¹For hydrogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 or the ring of substituted or unsubstituted C3-C6 Alkyl；

R²For nothing, halogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 or substituted or unsubstituted C3-C6 Cycloalkyl；

Y¹、Y²Independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylidene, replacement or unsubstituted Propylidene or substituted or unsubstituted butylidene；

R³For hydrogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 ,-C (=O) R⁴、-SO₂R⁵Replace or not The cycloalkyl of substituted C3-C6, wherein, R⁴、R⁵Independently be the straight or branched alkane of substituted or unsubstituted C1-C4 Base, the cycloalkyl of substituted or unsubstituted C3-C6 or C6-C10 aryl；

X is CR⁶Or N, wherein R⁶For hydrogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 or replacement or not The cycloalkyl of substituted C3-C6；

Wherein said each replacement refers to have 1-3 the substituent group selected from lower group independently: hydroxyl, halogen, C1-C6 straight chain or Branched alkyl, C1-C4 straight or branched alkoxyl, the cycloalkyl of C3-C6, C6-C10 aryl, carboxyl.

In another preference, R¹For hydrogen, the straight or branched alkyl of C1-C4 or the cycloalkyl of C3-C6.

In another preference, R²For hydrogen, fluorine, chlorine, bromine, the straight or branched alkyl of C1-C4 or the ring of C3-C6 Alkyl.

In another preference, R⁴、R⁵Independently be the straight or branched alkyl of substituted or unsubstituted C1-C4, C3-C6 Cycloalkyl or phenyl, described replacement refers to have 1-2 the substituent group selected from lower group: hydroxyl, halogen, C1-C4 straight chain Or branched alkoxy.

In another preference, R⁶For hydrogen, the straight or branched alkyl of C1-C4 or the cycloalkyl of C3-C6.

In another preference, Y¹For substituted or unsubstituted methylene, Y²Substituted or unsubstituted ethylidene；Or Y¹ For substituted or unsubstituted ethylidene, Y²For substituted or unsubstituted methylene, wherein

Described each replacement refers to have 1-2 the substituent group selected from lower group independently: hydroxyl, halogen, C1-C4 straight or branched Alkyl, C1-C4 straight or branched alkoxyl, the cycloalkyl of C3-C6.

In another preference, described compound is the arbitrary compound in I-1 to I-33 prepared by embodiment.

A second aspect of the present invention, it is provided that the preparation method of the compound described in first aspect, including the change described in formula II Compound generation ammonolysis obtains the step of the compound described in formula I:

Wherein, R¹、R²、Y¹、Y²、R³, X definition as described in relation to the first aspect.

A third aspect of the present invention, it is provided that a kind of pharmaceutical composition, including the compound described in first aspect, R-isomer, S-isomer or its pharmaceutically acceptable salt；And

Pharmaceutically acceptable carrier.

A fourth aspect of the present invention, it is provided that the compound described in first aspect or the use of the pharmaceutical composition described in the third aspect On the way, it is used for preparing:

(1) PARP1 inhibitor；

(2) PARP2 inhibitor；

(3) prevent and/or treat the medicine of tumor；

(4) anti-inflammatory drug；And/or

(5) prevent and/or treat the medicine with PARP relevant disease.

A fifth aspect of the present invention, it is provided that the intermediate of the compound described in a kind of first aspect, structure is as shown in general formula III:

Wherein,

R¹⁰For hydrogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 or the ring of substituted or unsubstituted C3-C6 Alkyl；

R⁹For nothing, halogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 or substituted or unsubstituted C3-C6 Cycloalkyl；

Y³、Y⁴Independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylidene, replacement or unsubstituted Propylidene or substituted or unsubstituted butylidene；

Y⁵For S or O；

R⁸For hydrogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 ,-C (=O) R¹¹、-SO₂R¹², replace or not The cycloalkyl of substituted C3-C6, wherein, R¹¹、R¹²Independently be the straight or branched of substituted or unsubstituted C1-C4 Alkyl, the straight or branched alkoxyl of substituted or unsubstituted C1-C4, the cycloalkyl of substituted or unsubstituted C3-C6 or C6-C10 aryl；

X is CR⁷Or N, wherein R⁷For hydrogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 or replacement or not The cycloalkyl of substituted C3-C6；

A sixth aspect of the present invention, it is provided that the intermediate of the compound described in a kind of first aspect, structure is as shown in formula IV:

Wherein, Y³、Y⁴Independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylidene, replacement or Unsubstituted propylidene or substituted or unsubstituted butylidene；

Y⁵For S or O；

R⁸For hydrogen, the straight or branched alkyl of substituted or unsubstituted C1-C4 ,-C (=O) R¹¹、-SO₂R¹², replace or not The cycloalkyl of substituted C3-C6, wherein, R¹¹、R¹²Independently be the straight or branched of substituted or unsubstituted C1-C4 Alkyl, the straight or branched alkoxyl of substituted or unsubstituted C1-C4, the cycloalkyl of substituted or unsubstituted C3-C6 or C6-C10 aryl；It is preferred that R⁸For-BOC i.e. tertbutyloxycarbonyl；

R¹³For hydrogen, halogen ,-CHO, the straight or branched alkyl of substituted or unsubstituted C1-C4 or replace or do not take The cycloalkyl of the C3-C6 in generation；It is preferred that be hydrogen, halogen or-CHO；

In should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and the most specifically retouching Can be combined with each other between each technical characteristic stated, thus constitute new or preferred technical scheme.As space is limited, at this Tire out the most one by one and state.

Detailed description of the invention

Present inventor, through extensively and in depth studying, has designed and synthesized the benzimidazole of a class sulfur atom-containing first -4-benzamide type PARP1/2 inhibitor.Compound involved in the present invention has clear and definite structure activity relationship, and shows very well Inside and outside anti-tumor activity.Such novel PARP1/2 inhibitor is expected to become new type antineoplastic medicine.On this basis Complete the present invention.

Term

In the context of the present invention, term " alkyl " represents saturated linear or branched chain hydrocarbon moiety, such as-CH₃Or -CH(CH₃)₂.Term " alkoxyl " represents-O-(alkyl) group.Term " cycloalkyl " represents saturated cyclic hydrocarbyl moiety, Such as cyclopropyl, cyclohexyl.Term " aryl " represents the hydrocarbyl portion comprising one or more aromatic ring.The example of aryl moiety Including phenyl (Ph), naphthyl etc..

Unless otherwise indicated, alkyl as herein described, alkoxyl, cycloalkyl and aryl include substituted and unsubstituted simultaneously Part.Substituent group possible on alkyl, alkoxyl, cycloalkyl and aryl includes, but are not limited to: C1-C6 alkyl, C2-C6 Thiazolinyl, C2-C6 alkynyl, C3-C10 cycloalkyl, C1-C20 Heterocyclylalkyl, C1-C10 alkoxyl, C6-C10 aryl, ammonia Base, C1-C10 alkyl amino, C1-C20 dialkyl amido, cyano group, nitro and carboxyl.

Compound shown in formula I

The compound shown in formula I that the present invention provides, structure is as follows:

R¹、R²、Y¹、Y²、R³, X as defined above shown in.

The compound of the present invention has asymmetric center, chiral axis and chiral planes, and can be with racemic modification, R-isomery Presented in body or S-isomer.Those skilled in the art can use routine techniques means to be obtained by racemate resolution R-isomer and/or S-isomer.

The present invention provide also compound of Formula I pharmaceutically can and the salt that accepts, in particular compound of Formula I and mineral acid Or organic acid reaction forms the pharmaceutically acceptable salt of routine.Such as, conventional pharmaceutically acceptable salt can pass through formula I prepares with mineral acid or organic acid reaction, and described mineral acid includes hydrochloric acid, hydrobromic acid, sulphuric acid, nitric acid, amido Sulfonic acid and phosphoric acid etc., and described organic acid includes citric acid, tartaric acid, lactic acid, acetone acid, acetic acid, benzenesulfonic acid, right Toluenesulfonic acid, methanesulfonic acid, LOMAR PWA EINECS 246-676-2, ethyl sulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, amber Amber acid, propanoic acid, oxalic acid, trifluoroacetic acid, stearic acid, flutter acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, Glutamic acid, ascorbic acid, para-anilinesulfonic acid, Aspirin and isethionic acid etc.；Or compound of Formula I Sodium salt, potassium salt, calcium salt, aluminium salt or the ammonium salt formed with inorganic base；Or the methylamine that compound of Formula I is formed with organic base Salt, ethylamine salt or ethanolamine salt.

Preparation method

(1) synthetic schemes of the compound that formula I-1, I-4, I-6～I-8, I-15, I-17～I-29 represent is as follows.

O-phenylenediamine compounds 1 and aldehyde derivative 2 contract in the presence of sodium sulfite or other lewis acids Cyclization also aoxidizes generation compound 3；Compound 3 generates compound 4 through de-Boc protection group in the presence of trifluoracetic acid, Ammonolysis generates compound I-1, I-4, I-6～I-8, I-15, I-17～I-29 again.

Wherein intermediate 1 can be prepared by commercially-available or following steps:

Isatin compounds 8 nitrification generates compound 9, and compound 9 occurs in the presence of peroxide such as hydrogen peroxide Baeyer-Villiger oxidation hydrolyzed under basic conditions open loop generate compound 10, under the catalysis of acid, and compound 10 ester Metaplasia becomes compound 11, and through palladium hydrocarbonization or other reducing agents such as iron powder, in the presence of zinc powder etc., reduction generates compound 1；

Aldehydes intermediate 2 is prepared by following steps:

Compound 12 obtains compound 2 under conditions of n-BuLi or tert-butyl lithium/dry DMF/anhydrous THF；

Compound 12 can be prepared by following steps:

Wherein intermediate 12-1 Yu 12-2 is by thieno tetrahydropyridines 13-1 Yu 13-2 bromo after boc protects Prepare；

Intermediate 12-3～12-10 is prepared by following steps:

Compound 15 through be acylated after at POCl₃Effect under cyclization obtain compound 17-1～17-8, then through reduction Boc Last bromo is protected to obtain compound 12-3～12-10；

Compound 12-11 is prepared by following steps:

Compound 18 generates compound 19 with nitroethane generation aldol condensation, through reducing such as lithium aluminium hydride reduction etc., then Boc Generate compound 21 after protection, then cyclization occurs in the presence of acid with paraformaldehyde, finally obtain with NBS bromo Compound 12-11；

Compound 12-12 is prepared by following steps:

Compound 22 and cyan-acetic ester, sulfur powder three component are heated to reflux generating compound 23, and compound 23 is through deamination Base/reduce two steps generate alcohol 25, deaminizating process can with but be not limited to amyl nitrite backflow and realize, reduction process can be used But being not limited to lithium borohydride backflow realize, compound 25 chloro in the presence of mesyl chloride or phosphorus oxychloride etc., subsequently at thunder In the presence of the metallic catalysts such as Buddhist nun's nickel, hydrogenation-dehalogenation element obtains compound 14-12, and last bromo obtains compound 12-12；

Compound 12-13 is prepared by following steps:

Compound 22 generates compound 27 with cyanamide, the cyclisation of sulfur powder, then obtains compound 12-13 through Sandmeyer reaction；

(2) synthetic schemes of the compound that formula I-2, I-3, I-5, I-9～I-14, I-16, I-17～I-29 represent is as follows.

There is alkylation or acylation or the sulfonylating compound 5 that obtains in compound 4, then ammonolysis generates compound I-2, I-3, I-5, I-9～I-14, I-16, I-17～I-29.

(3) synthetic schemes of the compound that I-30～I-33 of the present invention represents is as follows.

Compound 3 occurs alkylation to obtain compound 6, generates compound 7 through de-Boc protection group, then ammonolysis generates chemical combination Thing I-30～I-33.

Wherein R¹, R², R³As defined above with X.

In the present invention, the reagent Chinese table that chemical formula or English alphabet abbreviation represent is as follows:

NaHSO₃: sodium sulfite；EtOH: ethanol；DCM: dichloromethane；TFA: trifluoroacetic acid；NH₃: ammonia Gas；MeOH: methanol；NaOH: sodium hydroxide；H₂O₂: hydrogen peroxide；TsOH·H₂O: one hydration p-methyl benzenesulfonic acid； Pd/C: palladium/charcoal；H₂: hydrogen；NBuLi: n-BuLi；TBuLi: tert-butyl lithium；DMF:N, N-dimethyl formyl Amine；THF: oxolane；(Boc)₂O: Bis(tert-butoxycarbonyl)oxide；Et₃N: triethylamine；NBS:N-bromo succinyl Imines；MeCN: acetonitrile；POCl₃: phosphorus oxychloride；NaBH₄: sodium borohydride；EtNO₂: nitroethane；LiAlH₄: Lithium aluminium hydride reduction；(CH₂O)_nParaformaldehyde；LiBH₄: lithium borohydride；MsCl: mesyl chloride；CuBr₂: copper bromide；K₂CO₃: Potassium carbonate；Cs₂CO₃: cesium carbonate.

Unless otherwise defined, the meaning phase that all specialties used in literary composition are familiar with one skilled in the art with scientific words With.Additionally, any method similar or impartial to described content and material all can be applicable in the inventive method.Institute in literary composition The preferable implementation stated only presents a demonstration with material and is used.

Pharmaceutical composition

Present invention also offers a kind of pharmaceutical composition, it comprises the active component in the range of safe and effective amount, and pharmaceutically Acceptable carrier.

" active component " of the present invention refers to the 2-substituted benzimidazole-4-Methanamide shown in formula I of the present invention Compounds.

2-substituted benzimidazole-4-the Carbox amide of the present invention and pharmaceutical composition, can serve as PARP1 inhibitor Or PARP2 inhibitor；May be used for preparation prevention and/or treat disease relevant to Poly adenosine diphosphate-ribose polymerase-1 PARP Sick medicine；May be used for preparation prevention and/or the medicine for the treatment of tumor；Can be also used for preparing anti-inflammatory drug.

In another preference, described with Poly adenosine diphosphate-ribose polymerase-1 PARP relevant disease be tumor, inflammation and The disease such as cardiovascular disease that ischemia-reperfusion is concurrent, diabetes, rheumatic arthritis, endotoxin shock, in Wind etc..

In another preference, described tumor is the tumor of homologous recombination repair defect, i.e. BRCA1 or BRCA2 lacks Or the tumor of sudden change, such as ovarian cancer, breast carcinoma, carcinoma of prostate, gastric cancer, cancer of pancreas, cervical cancer, glioma, outstanding Wen's sarcoma etc..

" safe and effective amount " refers to: the amount of active component be enough to be obviously improved the state of an illness, and is unlikely to produce serious side effect. Generally, pharmaceutical composition contains 1-2000mg active component/agent, more preferably, containing 10-200mg active component/agent.Relatively Goodly, described " potion " is a tablet.

" pharmaceutically acceptable carrier " refers to: one or more biocompatible solid or liquid filler or gelatinous mass, they It is suitable for people to use and it is necessary to have enough purity and of a sufficiently low toxicity." compatibility " referred to herein as in compositions respectively Component energy and the active component of the present invention and they between mutually admix, and significantly reduce the drug effect of active component.Pharmacy Upper acceptable carrier part example has cellulose and its derivates (such as sodium carboxymethyl cellulose, ethyl cellulose sodium, fibre Dimension element acetas etc.), gelatin, Talcum, kollag (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil is (such as bean Oil, Oleum sesami, Oleum Arachidis hypogaeae semen, olive oil etc.), polyhydric alcohol (such as propylene glycol, glycerol, mannitol, sorbitol etc.), emulsifying agent (such as tween), wetting agent (such as sodium lauryl sulphate), coloring agent, flavoring agent, stabilizer, antioxidant, preservative, Apirogen water etc..

In another preference, the 2-substituted benzimidazole-4-Carbox amide shown in formula I of the present invention can be with big point Sub-compound or macromolecule form complex by nonbonding effect.In another preference, shown in formula I of the present invention 2-substituted benzimidazole-4-Carbox amide as little molecule also by chemical bond and macromolecular compound or macromolecule It is connected.Described macromolecular compound can be the highest polysaccharide of biomacromolecule, albumen, nucleic acid, polypeptide etc..

The active component of the present invention or the method for application of pharmaceutical composition are not particularly limited, and representational method of application includes (but being not limited to): be administered orally, tumor is interior, rectum, parenteral (intravenous, intramuscular or subcutaneous) etc..

Solid dosage forms for oral administration includes capsule, tablet, pill, powder and granule.

In these solid dosage formss, active component mixes, such as sodium citrate with at least one conventional inert excipients (or carrier) Or dicalcium phosphate, or mix with following compositions: (a) filler or bulking agent, such as, starch, lactose, sucrose, glucose, Mannitol and silicic acid；(b) binding agent, such as, hydroxymethyl cellulose, alginate, gelatin, polyvinyl pyrrolidone, Sucrose and arabic gum；(c) wetting agent, such as, glycerol；(d) disintegrating agent, such as, agar, calcium carbonate, Rhizoma Solani tuber osi Starch or tapioca, alginic acid, some composition silicate and sodium carbonate；(e) retarding solvent, such as paraffin；F () absorbs Accelerator, such as, quaternary ammonium compound；(g) wetting agent, such as spermol and glyceryl monostearate；(h) adsorbent, Such as, Kaolin；(i) lubricant, such as, Talcum, calcium stearate, magnesium stearate, solid polyethylene glycol, 12 Alkyl sodium sulfate, or its mixture.In capsule, tablet and pill, dosage form also can comprise buffer agent.

Described solid dosage forms also can use coating and shell material to prepare, such as casing and other material well known in the art.They can Comprise opacifying agent, and, in this compositions, the release of active component can a certain portion in digestive tract in a delayed fashion Release in point.The example of adoptable embedding component is polymeric material and Wax.

Liquid formulation for oral administration includes pharmaceutically acceptable emulsion, solution, suspension, syrup or tincture.Remove Outside active component, liquid dosage form can comprise the conventional inert diluent used in this area, such as water or other solvent, solubilising Agent and emulsifying agent, example knows, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3 butylene glycol, dimethyl Methanamide and oil, particularly Oleum Gossypii semen, Oleum Arachidis hypogaeae semen, maize embryo oil, olive oil, Oleum Ricini and Oleum sesami or these materials Mixture etc..In addition to these inert diluents, compositions also can comprise auxiliary agent, as wetting agent, emulsifying agent and suspending agent, Sweeting agent, correctives and spice.

In addition to active component, suspension can comprise suspending agent, such as, ethoxylation isooctadecane alcohol, polyoxyethylene Alcohol and Isosorbide Dinitrate, microcrystalline Cellulose, aluminium methoxide and agar or the mixture etc. of these materials.

Compositions for parenteral injection can comprise physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, suspension Liquid or emulsion, and for being again dissolved into the sterilized powder of aseptic Injectable solution or dispersion liquid.Suitable is aqueous and non-aqueous Carrier, diluent, solvent or excipient include water, ethanol, polyhydric alcohol and suitable mixture thereof.

The compounds of this invention can be individually dosed, or with other treatment medicine (such as chemotherapeutic) administering drug combinations.

When making pharmaceutical composition, be the compounds of this invention of safe and effective amount is applicable to treatment mammal (as People), when wherein using, dosage is the effective dosage pharmaceutically thought, for the people of 60kg body weight, day is to medicament Amount is usually 1～2000mg, preferably 20～500mg.Certainly, concrete dosage is it is also contemplated that route of administration, patient health The factors such as situation, within the scope of these are all skilled practitioners technical ability.

Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are merely to illustrate this Invention rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in the following example, generally According to normal condition such as Sambrook et al., molecular cloning: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or according to the condition proposed by manufacturer.Unless Additionally illustrating, otherwise percentage ratio and number are calculated by weight.

In all embodiments,¹H NMR is by Varian Mercury-300 or Varian Mercury-400 type nuclear magnetic resonance analyser Record,¹³C NMR is by Varian Mercury-400 or Varian Mercury-500 type or Varian Mercury-600 type Nuclear magnetic resonance analyser record, chemical shift represents with δ (ppm)；Mass spectrum is by Finnigan/MAT-95 (EI) and Finnigan LCQ/DECA and Micromass Ultra Q-TOF (ESI) type mass spectrograph record；Separation silica gel is 200-300 mesh.

Embodiment 1. 2-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, I-1's Preparation:

Step 1. 6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

Weighing 4,5,6,7-Tetramethylene sulfides [3,2-c] pyridine hydrochloride (13-1) (10g, 56.9mmol) is in reaction bulb (500mL) In, add methylene chloride (300mL).Add triethylamine (8.7mL) under ice bath, add Bis(tert-butoxycarbonyl)oxide (13.7 lentamente G, 62.8mmol) dichloromethane (20mL) solution.After dropping, within 1 hour, following the tracks of reaction, raw material reaction is complete Entirely.Reactant liquor is washed with water successively, saturated NaCl solution washing, retain organic layer, use anhydrous Na₂SO₄It is dried, Filtering concentrating under reduced pressure, thick product obtains water white transparency oily thing through column chromatography (petroleum ether: ethyl acetate=20:1), solid afterwards Turning to white solid, productivity is 99%.

¹H NMR (300MHz, Chloroform-d) δ 7.12 (d, J=5.1Hz, 1H), 6.78 (d, J=5.1Hz, 1H), 4.50 (s, 2H), 3.71 (t, J=5.3Hz, 2H), 2.84 (t, J=5.3Hz, 2H), 1.48 (s, 9H)；MS(ESI):m/z 240[M+H]⁺。

Step 2. 2-bromo-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

To 6, the acetonitrile of 7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (14-1) (2.39g, 10mmol) (40mL) solution is slowly added to N-bromo-succinimide (1.87g, 10.5mmol), continues stirring 1 hour. Reactant liquor is extracted with ethyl acetate (50mL × 3), and organic layer with water, saturated aqueous common salt washing, retains organic layer successively, Being dried with anhydrous sodium sulfate, filter concentrating under reduced pressure, thick product obtains nothing through column chromatography (petroleum ether: ethyl acetate=20:1) Color clear oil thing, productivity is 92%.

¹H NMR (300MHz, Chloroform-d) δ 6.74 (s, 1H), 4.41 (s, 2H), 3.69 (t, J=5.7Hz, 2H), 2.73 (t, J=5.8Hz, 2H), 1.48 (s, 9H)；MS(ESI):m/z 318[M+H]⁺。

Step 3. 2-formoxyl-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

-78 DEG C, under nitrogen protection, toward 2-bromo-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (12-1) Anhydrous tetrahydro furan (30mL) solution of (1g, 3.75mmol) is slowly added dropwise the tetrahydrochysene furan of 1.8mL n-BuLi Mutter solution (2.5M), continue stirring 1 hour.The anhydrous DMF of 330mg is added in reactant liquor, After stirring 1 hour, add 1mL shrend and go out.Reactant liquor is extracted with ethyl acetate (50mL × 3), and organic layer is used successively Washing, saturated common salt is washed, and retains organic layer, is dried with anhydrous sodium sulfate, filters concentrating under reduced pressure, and thick product is through column chromatography (petroleum ether: ethyl acetate=10:1) obtains water white transparency oily thing 841mg, and productivity is 84%.

¹H NMR (300MHz, Chloroform-d) δ 9.82 (s, 1H), 7.47 (s, 1H), 4.52 (s, 2H), 3.73 (t, J= 5.5Hz, 2H), 2.91 (t, J=5.4Hz, 2H), 1.49 (s, 9H)；MS(ESI):m/z 268[M+H]⁺。

Step 4. 2-(4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)- Carboxylic acid tert-butyl ester:

Toward 2-formoxyl-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (2-1) (830mg, 3.1mmol) Ethanol (15mL) solution in add 2,3-diamino-methyl benzoate (1-1) (540mg, 3.1mmol) and NaHSO₃(810mg, 7.8mmol), is heated to 60 DEG C, and reaction is overnight.Decompression boils off major part ethanol, residue Being extracted with ethyl acetate (50mL × 3), organic layer washes with water successively, and saturated common salt is washed, and retains organic layer, with anhydrous Sodium sulfate is dried, and filters concentrating under reduced pressure, and thick product obtains yellow powder through column chromatography (petroleum ether: ethyl acetate=4:1) 1.26g, productivity is 98%.

¹H NMR(300MHz,DMSO-d₆) δ 12.44 (s, 1H), 8.04 (s, 1H), 7.87 (d, J=7.8Hz, 1H), 7.80 (d, J=7.8Hz, 1H), 7.27 (t, J=7.8Hz, 1H), 4.47 (s, 2H), 3.96 (s, 3H), 3.66 (t, J=5.9 Hz, 2H), 2.84 (t, J=5.9Hz, 2H), 1.42 (s, 9H)；MS(ESI)414[M+H]⁺。

Step 5. 2-(4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

By 2-(4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid uncle Butyl ester (3-1) (4.6g, 11.1mmol) is dissolved in dichloromethane (20mL), is slowly added dropwise trifluoroacetic acid (5mL), It is stirred at room temperature 1 hour.Decompression boils off major part solvent, adds saturated Na in residue₂CO₃Solution (10mL), Being extracted with ethyl acetate (50mL × 3), organic layer washes with water successively, and saturated common salt is washed, and retains organic layer, with anhydrous Sodium sulfate is dried, and filters concentrating under reduced pressure, and thick product obtains yellow foam through column chromatography (dichloromethane: methanol=50:1) Solid 3.0g, productivity is 87%.

¹H NMR(300MHz,DMSO-d₆)δ12.63(brs,1H),9.24(brs,1H),8.07(s,1H),7.88(d,J =7.9Hz, 1H), 7.81 (d, J=7.8Hz, 1H), 7.30 (t, J=7.8Hz, 1H), 4.27 (s, 2H), 3.95 (s, 3H), 3.47 (t, J=5.9Hz, 2H), 3.11 (t, J=5.4Hz, 2H)；MS(ESI)314[M+H]⁺。

Step 6. 2-(4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide (I-1):

To 2-(4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate (4-1) (110 Mg, 0.35mmol) in add the saturated methanol solution (10mL) of ammonia, 80 DEG C of tube sealing reactions 20 hours.Will be anti- Answering liquid to be cooled to room temperature, decompression boils off major part solvent, and thick product obtains through column chromatography (dichloromethane: methanol=10:1) White powder 77mg, productivity is 74%.

¹H NMR(300MHz,DMSO-d₆) δ 9.09 (brs, 1H), 7.83 (dd, J=7.6,1.1Hz, 1H), 7.73 (brs, 1H), 7.68 (dd, J=8.0,1.1Hz, 1H), 7.63 (s, 1H), 7.31 (t, J=7.8Hz, 1H), 3.81 (s, 2H), 3.00 (t, J=5.6Hz, 2H), 2.76 (t, J=5.6Hz, 2H)；¹³C NMR(125MHz,DMSO-d₆)δ166.75,148.54, 138.68,136.96,135.53,128.84,126.66,126.66,123.10,122.49,122.04,115.62,45.30,43.40, 26.09；HRMS(ESI):m/z Calcd.For C₁₅H₁₅N₄OS[M+H]⁺:299.0967；Found:299.0959.

Embodiment 2. 2-(5-methyl-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, The preparation of I-2:

Step 1. 2-(5-methyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

By 2-(4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate (4-1) (198 Mg, 0.63mmol) it is dissolved in formic acid (3mL), add formalin (37%, 0.3mL).Reactant liquor is risen to 95 At DEG C react 4 hours complete to raw material reaction.Decompression boils off major part solvent, adds Na in residue₂CO₃Saturation water Solution regulation pH, to alkalescence, is extracted with ethyl acetate (50mL × 3), and organic layer washs with water, saturated aqueous common salt successively, Retain organic layer, be dried with anhydrous sodium sulfate, filter concentrating under reduced pressure, thick product through column chromatography (dichloromethane: methanol=80: 1) obtaining yellow bubble solid 128mg, productivity is 62%.

¹H NMR(300MHz,DMSO-d₆) δ 7.79 (s, 1H), 7.77 (d, J=7.1Hz, 1H), 7.69 (d, J=7.0 Hz, 1H), 7.16 (t, J=7.5Hz, 1H), 3.91 (s, 3H), 3.42 (s, 2H), 2.84 (t, J=5.9Hz, 2H), 2.66 (t, J=5.9Hz, 2H), 2.36 (s, 3H)；MS(ESI)328[M+H]⁺。

Step 2. 2-(5-methyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with 2-(5-methyl-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (5-1) (128mg, 0.39mmol) is that raw material ammonia solution obtains white powder 94mg, productivity is 77% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆) δ 9.49 (brs, 1H), 7.74 (dd, J=7.5,1.2Hz, 1H), 7.65 (dd, J=7.9,1.2Hz, 1H), 7.59 (brs, 1H), 7.57 (s, 1H), 7.18 (t, J=7.7Hz, 1H), 3.47 (s, 2H), 2.90 (t, J=5.6Hz, 2H), 2.71 (t, J=5.6Hz, 2H), 2.41 (s, 3H)；¹³C NMR(125MHz,DMSO-d₆)δ 167.58,151.40,143.15,140.55,135.75,135.59,133.04,125.06,121.69,121.20,120.59,116.90, 54.71,52.43,45.66,25.74；HRMS(ESI):m/z Calcd.For C₁₆H₁₇N₄OS[M+H]⁺:313.1123； Found:313.1114。

Embodiment 3. 2-(5-ethyl-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, The preparation of I-3:

Step 1. 2-(5-ethyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

By 2-(4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate (4-1) (160 Mg, 0.51mmol) it is dissolved in dry DMF (5mL), add Cs₂CO₃(183mg, 0.56mmol), bromine second After alkane (67mg, 0.61mmol), it is warming up at 80 DEG C reaction 1 hour.Reactant liquor is poured into water, ethyl acetate Extraction (50mL × 3), organic layer washes with water successively, and saturated common salt is washed, and retains organic layer, is dried with anhydrous sodium sulfate, Filtering concentrating under reduced pressure, thick product obtains light yellow gum thing 150mg through column chromatography (dichloromethane: methanol=80:1), Productivity is 86%.

¹H NMR(300MHz,DMSO-d₆) δ 12.42 (brs, 1H), 7.97 (s, 1H), 7.86 (d, J=7.9Hz, 1H), 7.78 (d, J=7.7Hz, 1H), 7.28 (t, J=7.8Hz, 1H), 3.96 (s, 3H), 3.54 (s, 2H), 2.86 (brs, 2H), 2.77 (brs, 2H), 2.58 (q, J=6.9Hz, 2H), 1.10 (t, J=7.1Hz, 3H)；MS(ESI)342[M+H]⁺。

Step 2. 2-(5-ethyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with 2-(5-ethyl-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2- Base) to be that raw material ammonia solution obtains light yellow solid for-1H-benzo [d] imidazoles-4-methyl formate (5-2) (150mg, 0.44mmol) Body 87mg, productivity is 61% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆) δ 13.44 (brs, 1H), 9.14 (brs, 1H), 7.84 (d, J=7.6Hz, 1 H), 7.78 (brs, 1H), 7.68 (d, J=8.0Hz, 1H), 7.64 (s, 1H), 7.32 (t, J=7.7Hz, 1H), 3.66 (s, 2 H), 2.92 (s, 2H), 2.89 (brs, 2H), 2.67 (brs, 2H), 1.14 (t, J=7.2Hz, 3H)；¹³C NMR(125MHz, DMSO-d₆)δ166.02,147.64,141.29,137.31,135.04,129.40,126.13,122.83,122.23,121.93, 114.62,51.36,50.76,49.46,24.89,11.86；HRMS(ESI):m/z Calcd.For C₁₇H₁₉N₄OS[M+H]⁺: 327.1280；Found:327.1273.

Embodiment 4. 2-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, I-4's Preparation:

Step 1. 4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 14-1, with 4,5,6,7-Tetramethylene sulfides [2,3-c] pyridine hydrochloride (13-2) (1.00 G, 5.7mmol) be raw material through protecting to obtain white solid 1.32g, productivity is 97% (petroleum ether: ethyl acetate=20:1).

¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.0Hz, 1H), 6.79 (d, J=5.1Hz, 1H), 4.62 (s, 2H), 3.67 (t, J=5.4Hz, 2H), 2.70 (t, J=5.4Hz, 2H), 1.8 (s, 9H)；MS(ESI)240 [M+H]⁺。

Step 2. 2-bromo-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 10-1, with 4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (14-2) (1.32g, 5.5mmol) is that raw material obtains water white transparency oily thing 1.65g through bromo, and productivity is 94% (oil Ether: ethyl acetate=20:1).

¹H NMR (300MHz, Chloroform-d) δ 6.74 (s, 1H), 4.50 (s, 2H), 3.64 (t, J=5.7Hz, 2H), 2.63 (t, J=5.8Hz, 2H), 1.47 (s, 9H)；MS(ESI):m/z 318[M+H]⁺。

Step 3. 2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-1, with 2-bromo-4,7-dihydro-thiophene also [2,3-c] pyridine-6 tertiary fourth of (5H)-carboxylic acid Ester (12-2) (1.65g, 5.2mmol) is that raw material prepares brown gum 1.12g, and productivity is 81% (oil Ether: ethyl acetate=10:1).

¹H NMR (300MHz, Chloroform-d) δ 9.82 (s, 1H), 7.47 (s, 1H), 4.67 (s, 2H), 3.68 (t, J= 5.6Hz, 2H), 2.75 (d, J=5.3Hz, 2H), 1.47 (s, 9H)；MS(ESI):m/z 268[M+H]⁺。

Step 4. 2-(4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid The tert-butyl ester (2-2) (1.12g, 4.2mmol) and 2,3-diamino-methyl benzoate (1-1) (0.70g, 4.2mmol) Preparing yellow powder 1.53g for raw material, productivity is 88% (petroleum ether: ethyl acetate=3:1).

¹H NMR (300MHz, Chloroform-d) δ 10.46 (brs, 1H), 7.95 (d, J=7.8Hz, 1H), 7.87 (d, J =7.7Hz, 1H), 7.37 (s, 1H), 7.29 (t, J=7.8Hz, 1H), 4.68 (brs, 2H), 4.01 (s, 3H), 3.73 (t, J= 5.6Hz, 2H), 2.76 (t, J=5.6Hz, 2H), 1.50 (s, 9H)；MS(ESI)414[M+H]⁺。

Step 5. 2-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with 2-(4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-bis- Hydrogen thieno [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-2) (1.53g, 3.7mmol) is that raw material deprotection base obtains Buff powder 0.93g, productivity is 80% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.46 (brs, 1H), 7.94 (d, J=8.0Hz, 1H), 7.85 (d, J =7.8Hz, 1H), 7.37 (s, 1H), 7.28 (t, J=7.9Hz, 1H), 4.08 (brs, 2H), 4.00 (s, 3H), 3.15 (t, J= 5.8Hz, 2H), 2.69 (t, J=5.8Hz, 2H)；MS(ESI)314[M+H]⁺。

Step 6. 2-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with 2-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate (4-2) (158mg, 0.50mmol) is that raw material ammonia solution obtains white solid 130mg, produces Rate is 87% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆) δ 9.12 (brs, 1H), 7.82 (d, J=7.5Hz, 1H), 7.75 (s, 1H), 7.69 (dd, J=7.9,0.9Hz, 1H), 7.66 (brs, 1H), 7.31 (t, J=7.8Hz, 1H), 3.92 (s, 2H), 2.96 (t, J =5.7Hz, 2H), 2.62 (t, J=5.4Hz, 2H)；¹³C NMR(125MHz,DMSO-d₆)δ166.71,148.52, 141.84,139.75,136.01,128.92,123.15,122.53,115.26,44.83,42.98,26.57；HRMS(ESI):m/z Calcd.For C₁₅H₁₅N₄OS[M+H]⁺:299.0967；Found:299.0959.

Embodiment 5. 2-(6-methyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, The preparation of I-5:

Step 1. 2-(6-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

According to preparing the operational approach of compound 5-1, with 2-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate (4-2) (158mg, 0.50mmol) is raw material, through methylate brown color cystose is solid Body 134mg, productivity is 82% (dichloromethane: methanol=30:1).

¹H NMR (400MHz, Chloroform-d) δ 10.47 (brs, 1H), 7.96 (d, J=7.9Hz, 1H), 7.88 (d, J =7.7Hz, 1H), 7.41 (s, 1H), 7.31 (t, J=7.8Hz, 1H), 4.04 (s, 3H), 3.71 (s, 2H), 2.87-2.82 (m, 2H),2.81-2.76(m,2H),2.54(s,3H)；MS(ESI)328[M+H]⁺。

Step 2. 2-(6-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with 2-(6-methyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base) to be that raw material ammonia solution obtains canescence solid for-1H-benzo [d] imidazoles-4-carboxylate methyl ester (5-3) (134mg, 0.41mmol) Body 102mg, productivity is 80% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆) δ 9.15 (brs, 1H), 7.84 (d, J=6.3Hz, 1H), 7.77 (brs, 1H), 7.69 (d, J=7.9Hz, 1H), 7.65 (brs, 1H), 7.32 (t, J=7.4Hz, 1H), 3.60 (s, 2H), 2.73 (t, J=5.0 Hz, 2H), 2.67 (t, J=5.5Hz, 2H), 2.39 (s, 3H)；¹³C NMR(125MHz,DMSO-d₆)δ166.54, 148.28,141.82,137.62,135.55,135.14,129.45,128.26,123.31,122.71,122.43,115.12,53.74, 52.01,45.42,25.67；HRMS(ESI):m/z Calcd.For C₁₆H₁₇N₄OS[M+H]⁺:313.1123；Found: 313.1118。

The chloro-2-of embodiment 6. 6-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, I-6 Preparation:

Step 1. 5-chloro-7-nitro indoline-2,3-diketone:

At-5 DEG C, slow in concentrated sulphuric acid (6mL) solution dissolved with 5-chlorisatide (8-1) (1.81g, 10mmol) Slowly dripping fuming nitric aicd (1mL), stir 1 hour, reactant liquor is poured in frozen water, separates out solid, filters washing, dry Dry yellow solid 1.77g, productivity is 78%.

¹H NMR(300MHz,DMSO-d₆) δ 11.80 (s, 1H), 8.32 (d, J=2.2Hz, 1H), 8.02 (d, J=2.3 Hz,1H),4.48(s,1H)；MS(ESI)227[M+H]⁺。

Step 2. 2-amino-5-chloro-3-nitrobenzoic acid:

At 0 DEG C, by chloro-for 5-7-nitro indoline-2,3-diketone (9-1) (1.77g, 7.8mmol) adds to sodium hydroxide Aqueous solution (5N, 15mL) is formed suspension, then is slowly added dropwise 30% hydrogenperoxide steam generator (2mL) in suspension, Move to room temperature after dropping and continue stirring 5 hours, be neutralized to pH=4.0 with 2N hydrochloric acid, separate out precipitation and filter, washing, Being dried to obtain yellow solid 1.17g, productivity is 69%.

¹H NMR(300MHz,DMSO-d₆) δ 13.92 (brs, 1H), 8.46 (brs, 2H), 8.28 (d, J=2.7Hz, 1 H), 8.11 (d, J=2.6Hz, 1H)；MS(ESI)217[M+H]⁺。

Step 3. 2-amino-5-chloro-3-nitrobenzene methyl:

In methanol (60mL) solution of 2-amino-5-chloro-3-nitrobenzoic acid (10-1) (1.17g, 5.38mmol) Add hydration p-methyl benzenesulfonic acid (1.02g, 5.38mmol), be heated to reflux 36 hours.Cooling reactant liquor, decompression is steamed Except solvent, thick product obtains yellow solid 0.53g through column chromatography (petroleum ether: ethyl acetate=8:1), and productivity is 43%.

¹H NMR (300MHz, Chloroform-d) δ 8.43 (brs, 2H), 8.38 (d, J=2.6Hz, 1H), 8.20 (d, J =2.6Hz, 1H), 3.93 (s, 3H)；MS(ESI)231[M+H]⁺。

Step 4. 2,3-diaminourea-5-chloro benzoic ether:

Methanol (30mL) toward 2-amino-5-chloro-3-nitrobenzene methyl (11-1) (530mg, 2.31mmol) Adding the palladium charcoal 50mg of 10% in solution, displacement nitrogen three times, then replacing hydrogen three times, in the nitrogen atmosphere of an atmospheric pressure Enclose lower stirring 5 hours, filter, decompression boil off major part solvent, thick product through column chromatography (petroleum ether: ethyl acetate=4: 1) obtaining pale solid 51mg, productivity is 11%.

¹H NMR (300MHz, Chloroform-d) δ 7.27 (d, J=1.9Hz, 1H), 6.70 (d, J=1.9Hz, 1H), 4.46(brs,4H),3.86(s,3H),2.21(s,3H)；MS(ESI)201[M+H]⁺。

Step 5. 2-(6-chloro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters:

According to preparing the operational approach of compound 3-1, with 2, and 3-diaminourea-5-chloro benzoic ether (1-2) (51mg, 0.25 And 2-formoxyl-4 mmol), and 7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (2-2) (67mg, 0.25 Mmol) being that raw material prepares pale yellow powder 73mg, productivity is 65% (petroleum ether: ethyl acetate=3:1).

¹H NMR (300MHz, Chloroform-d) δ 10.49 (brs, 1H), 7.88 (d, J=1.8Hz, 1H), 7.82 (d, J =1.7Hz, 1H), 7.38 (s, 1H), 4.67 (s, 2H), 4.00 (s, 3H), 3.71 (t, J=5.0Hz, 2H), 2.75 (t, J= 5.3Hz,2H),1.50(s,9H)；MS(ESI)448[M+H]⁺。

The chloro-2-of step 6. 6-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with 2-(6-chloro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7- Dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-3) (73mg, 0.16mmol) is that raw material deprotection base obtains To light brown powder 48mg, productivity is 86% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.50 (brs, 1H), 7.89 (d, J=1.9Hz, 1H), 7.82 (d, J =1.9Hz, 1H), 7.41 (s, 1H), 6.86 (s, 1H), 4.10 (s, 2H), 4.02 (s, 3H), 3.16 (t, J=5.8Hz, 2H), 2.72 (t, J=5.6Hz, 2H)；MS(ESI)348[M+H]⁺。

The chloro-2-of step 7. 6-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with the chloro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-3) (48mg, 0.14mmol) is that raw material ammonia solution obtains Light brown solid 30mg, Productivity is 65% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ8.96(brs,1H),7.94(brs,1H),7.83(s,1H),7.74(d,J =2.2Hz, 1H) 7.73 (d, J=2.2Hz, 1H), 4.13 (s, 2H), 3.15 (t, J=6.2Hz, 2H), 2.76 (t, J=6.2 Hz,2H)；¹³C NMR(125MHz,DMSO-d₆)δ165.49,149.53,136.28,135.26,129.80,129.21, 126.88,122.73,115.09,43.52,41.95,29.47；HRMS(ESI):m/z Calcd.For C₁₅H₁₄ClN₄OS [M+H]⁺:333.0577；Found:333.0570.

Embodiment 7. 6-methyl-2-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, The preparation of I-7:

Step 1. 5-methyl-7-nitro indoline-2,3-diketone:

According to preparing the operational approach of compound 9-1, with 5-methylisatin (8-2) (2.49g, 20mmol) for raw material, Nitrification prepares yellow solid 1.50g, and productivity is 37%.

¹H NMR(300MHz,DMSO-d₆)δ11.58(brs,1H),8.13(s,1H),7.77(s,1H),2.35(s,3 H)；MS(ESI)207[M+H]⁺。

Step 2. 2-amino-5-methyl-3-nitro benzoic acid:

According to preparing the operational approach of compound 10-1, with 5-methyl-7-nitro indoline-2,3-diketone (9-2) (1.5g, Being 7.4mmol) that raw material prepares yellow solid 1.23g, productivity is 85%.

¹H NMR(300MHz,DMSO-d₆)δ13.50(brs,1H),8.33(brs,2H),8.14(s,1H),8.07(s,1 H),2.24(s,3H)；MS(ESI)197[M+H]⁺。

Step 3. 2-amino-5-methyl-3-nitro essence of Niobe:

According to preparing the operational approach of compound 11-1, with 2-amino-5-methyl-3-nitro benzoic acid (10-2) (1.23g, Being 6.3mmol) that raw material prepares yellow solid 0.50g, productivity is 38% (petroleum ether: ethyl acetate=8:1).

¹H NMR(300MHz,Chloroform-d)δ8.21(s,1H),8.08(s,1H),7.71(brs,2H),3.91(s,3 H),2.29(s,3H)；MS(ESI)211[M+H]⁺。

Step 4. 2,3-diaminourea-5-methyl toluate:

According to preparing the operational approach of compound 1-2, with 2-amino-5-methyl-3-nitro essence of Niobe (11-2) (500 Mg, 2.4mmol) it is that raw material prepares brown solid 90mg, productivity is 21% (petroleum ether: ethyl acetate=4:1).

¹H NMR (300MHz, Chloroform-d) δ 7.27 (d, J=1.9Hz, 1H), 6.70 (d, J=1.9Hz, 1H), 4.46(brs,4H),3.86(s,3H),2.21(s,3H)；MS(ESI)181[M+H]⁺。

Step 5. 2-(4-(methoxycarbonyl)-6-methyl isophthalic acid H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters:

According to preparing the operational approach of compound 3-1, with 2,3-diaminourea-5-methyl toluate (1-3) (90mg, 0.5mmol) with 2-formoxyl-4, and 7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (2-2) (134mg, 0.5 Mmol) being that raw material prepares pale yellow powder 96mg, productivity is 45% (petroleum ether: ethyl acetate=3:1).

¹H NMR(300MHz,Chloroform-d)δ10.40(brs,1H),7.72(s,1H),7.69(s,1H),7.35(s, 1H), 4.66 (s, 2H), 3.98 (s, 3H), 3.70 (t, J=5.3Hz, 2H), 2.73 (t, J=4.9Hz, 2H), 2.48 (s, 3 H),1.49(s,9H)；MS(ESI)428[M+H]⁺。

Step 6. 6-methyl-2-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with 2-(4-(methoxycarbonyl)-6-methyl isophthalic acid H-benzo [d] imidazoles-2- Base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-4) (96mg, 0.23mmol) is that raw material goes Protection group obtains light brown powder 59mg, and productivity is 79% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.34 (s, 1H), 7.74 (dd, J=1.5,0.8Hz, 1H), 7.69 (dd, J=1.5,0.7Hz, 1H), 7.36 (s, 1H), 4.09 (s, 2H), 4.00 (s, 3H), 3.16 (t, J=5.8Hz, 2H), 2.70 (t, J=5.9Hz, 2H), 2.50 (s, 3H)；MS(ESI)328[M+H]⁺。

Step 7. 6-methyl-2-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with 6-methyl-2-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-carboxylate methyl ester (4-4) (59mg, 0.15mmol) is that raw material ammonia solution obtains brown solid 48 Mg, productivity is 67% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ9.09(brs,1H),7.79(s,1H),7.75(brs,1H),7.64(s,1 H), 7.49 (s, 1H), 4.22 (s, 2H), 3.22 (t, J=5.8Hz, 2H), 2.84 ((t, J=5.8Hz, 2H), 2.44 (s, 3H)；¹³C NMR(125MHz,DMSO-d₆)δ166.75,147.45,139.87,135.96,134.71,133.37,132.29, 130.91,128.18,124.58,121.66,115.19,42.77,41.46,23.69,21.67；HRMS(ESI):m/z Calcd. For C₁₆H₁₇N₄OS[M+H]⁺:313.1123；Found:313.1119.

The fluoro-2-of embodiment 8. 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, I-8 Preparation:

Step 1. 5-fluoro-7-nitro indoline-2,3-diketone:

According to preparing the operational approach of compound 9-1, with 5-fluoro indigo red (8-3) (10.0g, 60.6mmol) for raw material, Nitrification prepares yellow solid 10.6g, and productivity is 83%.

¹H NMR(300MHz,DMSO-d₆) δ 11.71 (s, 1H), 8.21 (dd, J=9.1,2.7Hz, 1H), 7.97 (dd, J=6.4,2.7Hz, 1H)；MS(ESI)211[M+H]⁺。

Step 2. 2-amino-5-fluorine-3-nitrobenzoic acid:

According to preparing the operational approach of compound 10-1, with 5-fluoro-7-nitro indoline-2,3-diketone (9-3) (10.6g, Being 50.0mmol) that raw material prepares yellow solid 7.0g, productivity is 70%.

¹H NMR(300MHz,DMSO-d₆) δ 8.33 (brs, 2H), 8.17 (dd, J=8.8,3.3Hz, 1H), 8.04 (dd, J=8.6,3.3Hz, 1H)；MS(ESI)201[M+H]⁺。

Step 3. 2-amino-5-fluorine-3-nitrobenzene methyl:

According to preparing the operational approach of compound 11-1, with 2-amino-5-fluorine-3-nitrobenzoic acid (10-3) (6.0g, 30.0 Mmol) being that raw material prepares yellow solid 2.5g, productivity is 39% (petroleum ether: ethyl acetate=4:1).

¹H NMR (300MHz, Chloroform-d) δ 8.31 (brs, 2H), 8.14 (dd, J=8.4,3.2Hz, 1H), 8.02 (dd, J=8.3,3.2Hz, 1H)；MS(ESI)215[M+H]⁺。

Step 3. 2,3-diaminourea-5-fluorophenyl carbamate:

According to preparing the operational approach of compound 1-2, with 2-amino-5-fluorine-3-nitrobenzene methyl (11-3) (2.5g, Being 11.7mmol) that raw material prepares pale solid 1.5g, productivity is 70% (petroleum ether: ethyl acetate=4:1).

¹H NMR (300MHz, Chloroform-d) δ 7.11 (dd, J=9.8,2.9Hz, 1H), 6.62 (dd, J=9.1,2.9 Hz,1H),5.32(brs,2H),3.87(s,3H),3.51(brs,2H)；MS(ESI):185[M+H]⁺。

Step 4. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-6,7-dihydro-thiophene also [3,2-c] pyridine -5 (4H)-carboxylic acid tert-butyl esters:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid The tert-butyl ester (2-1) (726mg, 2.72mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (500mg, Being 2.72mmol) that raw material prepares white powder 1.06g, productivity is 90% (petroleum ether: ethyl acetate=3:1).

¹H NMR (400MHz, Chloroform-d) δ 10.40 (brs, 1H), 7.63 (dd, J=8.8,2.4Hz, 1H), 7.59 (dd, J=9.6,2.4Hz, 1H), 7.38 (s, 1H), 4.55 (s, 2H), 4.02 (s, 3H), 3.77 (t, J=5.1Hz, 2H), 2.92 (t, J=5.2Hz, 2H), 1.50 (s, 9H)；MS(ESI):432[M+H]⁺。

The fluoro-2-of step 5. 6-(4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-6,7- Dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (3-5) (1.0g, 2.3mmol) is that raw material deprotection base obtains Yellow foamy solid 670mg, productivity is 88% (dichloromethane: methanol=50:1).

¹H NMR(300MHz,DMSO-d₆) δ 7.97 (s, 1H), 7.74 (d, J=9.2Hz, 1H), 7.54 (d, J=9.2 Hz, 1H), 3.99 (s, 3H), 3.82 (s, 2H), 3.00 (t, J=4.8Hz, 2H), 2.76 (t, J=4.8Hz, 2H)；MS (ESI):332[M+H]⁺。

The fluoro-2-of step 6. 6-(4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-5) (100mg, 0.30mmol) is that raw material ammonia solution obtains brown solid 78mg, Productivity is 82% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ9.02(brs,1H),7.88(brs,1H),7.61(s,1H),7.52(dd, J=10.0,2.5Hz, 1H), 7.50 (dd, J=8.0,2.5Hz, 1H), 3.79 (s, 2H), 2.99 (t, J=5.2Hz, 2H), 2.74 (t, J=5.2Hz, 2H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.69,158.58 (d, J=236.0Hz, C-F), 149.91,138.63,137.79,136.69,128.92,126.80,122.55,110.05 (d, J=26.5Hz, CH-CF), 102.57 (d, J=25.2Hz, CH-CF), 79.63,45.19,43.30,25.93；HRMS(ESI):m/z Calcd.For C₁₅H₁₄FN₄OS[M+H]⁺:317.0872；Found:317.0867.

The fluoro-2-of embodiment 9. 6-(5-methyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-9:

The fluoro-2-of step 1. 6-(5-methyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 5-1, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-5) (198mg, 0.60mmol) is raw material, through the yellow solid 200 that methylates to obtain Mg, productivity is 97% (dichloromethane: methanol=30:1).

¹H NMR (400MHz, Chloroform-d) δ 7.63 (dd, J=8.9,2.4Hz, 1H), 7.58 (dd, J=9.7,2.4 Hz, 1H), 7.34 (s, 1H), 4.02 (s, 3H), 3.55 (s, 2H), 2.99 (t, J=5.7Hz, 2H), 2.79 (t, J=5.7Hz, 2H),2.51(s,3H)；MS(ESI)346[M+H]⁺。

The fluoro-2-of step 2. 6-(5-methyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(5-methyl-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (4-5) (100mg, 0.29mmol) is that raw material ammonia solution obtains yellow powder End 58mg, productivity is 61% (dichloromethane: methanol=10:1).

¹H NMR(400MHz,DMSO-d₆)δ9.08(brs,1H),7.95(s,1H),7.65(s,1H),7.58–7.50 (m, 2H), 3.57 (s, 1H), 2.92 (t, J=5.1Hz, 2H), 2.79 (t, J=5.0Hz, 2H), 2.45 (s, 3H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.35,158.78 (d, J=237.0Hz, C-F), 149.18,138.48,137.68, (136.04,135.24,129.62,126.82,123.20,110.38 d, J=24.0Hz, CH-CF), 101.78 (d, J=25.0 Hz,CH-CF),54.18,52.00,45.11,25.31；HRMS(ESI):m/z Calcd.For C₁₆H₁₆FN₄OS[M+H]⁺: 331.1029；Found:331.1022.

Embodiment 10. 2-(5-acetyl group-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methanamide, the preparation of I-10:

Step 1. 2-(5-acetyl group-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Acid methyl ester:

At 0 DEG C, to dissolved with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester (4-5) (198mg, 0.60mmol) and the dry DMF (4mL) of triethylamine (121mg, 1.20mmol) Solution is slowly added dropwise chloroacetic chloride (57mg, 0.72mmol), after 30 minutes, reactant liquor is poured in mixture of ice and water, Separating out white solid, filter, be vacuum dried to obtain 197mg, productivity is 88%.

¹H NMR (400MHz, Chloroform-d) conformer 1: δ 10.50 (brs, 1H), 7.65 7.57 (m, 2H), (7.40 s, 1H), 4.71 (s, 2H), 4.02 (s, 3H), 3.79 (t, J=5.7Hz, 2H), 2.99 (t, J=5.6Hz, 2H), 2.22 (s,3H)；Conformer 2: δ 10.43 (brs, 1H), 7.65 7.57 (m, 2H), 7.40 (s, 1H), 4.59 (s, 2H), 4.02 (s, 3H), 3.96 (t, J=5.7Hz, 2H), 2.93 (t, J=5.6Hz, 2H), 2.20 (s, 3H)；MS(ESI)374 [M+H]⁺。

Step 2. 2-(5-acetyl group-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Amide

According to preparing the operational approach of compound I-1, with 2-(5-acetyl group-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2- Base)-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (5-6) (197mg, 0.53mmol) is that raw material ammonia solution obtains white Powder 182mg, productivity is 96% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆) conformer 1: δ 9.05 (brs, 1H), 7.92 (s, 1H), 7.71 (brs, 1H),7.60–7.50(m,2H),4.63(s,2H),3.75(m,2H),2.82(brs,2H),2.10(s,3H)；Conformational isomerism Body 2: δ 9.05 (brs, 1H), 7.92 (s, 1H), 7.71 (brs, 1H), 7.60 7.50 (m, 2H), 4.58 (s, 2H), 7.82–7.71(m,2H),2.95(brs,2H),2.12(s,3H)；¹³C NMR(125MHz,DMSO-d₆) conformational isomerism Body 1: δ 169.44,165.52,158.68 (d, J=229.0Hz, C-F), 149.23,138.18,137.75,134.39,134.06, 130.01,126.65 (d, J=25.2Hz, CH-CF), 110.38 (d, J=25.2Hz, CH-CF), 45.90,43.75,25.04, 22.31；Conformer 2: δ 169.38,165.52,158.68 (d, J=229.0Hz, C-F), 149.23,138.18,137.75, (134.39,134.06,130.01,126.65 d, J=25.2Hz, CH-CF), 110.38 (d, J=25.2Hz, CH-CF), 45.90,41.93,25.77,21.87；HRMS(ESI):m/z Calcd.For C₁₇H₁₆FN₄O₂S[M+H]⁺:359.0978； Found:359.0973。

Embodiment 11. 2-(5-(cyclopropyl carbonyl)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] Imidazoles-4-Methanamide, the preparation of I-11:

Step 1. 2-(5-(cyclopropyl carbonyl)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles -4-methyl formate:

According to preparing the operational approach of compound I-11, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-5) (182mg, 0.60mmol) and Cyclopropyl carbonyl chloride (75mg, 0.72mmol) Preparing white powder 182mg for raw material, productivity is 76%.

¹H NMR(300MHz,CD₃OD) conformer 1: δ 7.89 (s, 1H), 7.73 (s, 1H), 7.59 (dd, J= 9.8,2.3Hz, 1H), 7.52 (dd, J=8.5,2.3Hz, 1H), 4.92 (s, 2H), 4.02 (s, 3H), 3.93 (t, J=5.9Hz, 2H), 2.90 (t, J=5.7Hz, 2H), 2.10 (m, 1H), 0.96 0.85 (m, 4H)；Conformer 2: δ 7.89 (s, 1 H), 7.73 (s, 1H), 7.59 (dd, J=9.8,2.3Hz, 1H), 7.52 (dd, J=8.5,2.3Hz, 1H), 4.68 (s, 2H), 4.09 (t, J=5.9Hz, 2H), 4.02 (s, 3H), 3.04 (t, J=5.8Hz, 2H), 2.10 (m, 1H), 0.96 0.85 (m, 4 H)；MS(ESI)400[M+H]⁺。

Step 2. 2-(5-(cyclopropyl carbonyl)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles -4-Methanamide:

According to preparing the operational approach of compound I-1, with 2-(5-(cyclopropyl carbonyl)-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine -2-base) to be that raw material ammonia solution obtains shallow for-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (5-7) (182mg, 0.46mmol) Brown solid 106mg, productivity is 60% (dichloromethane: methanol=10:1).

¹H NMR(400MHz,DMSO-d₆) conformer 1: δ 9.06 (brs, 1H), 7.94 (s, 1H), 7.72 (brs, 1H),7.60–7.52(m,2H),4.88(s,2H),3.82(brs,2H),2.83(brs,2H),2.10(m,1H),0.75(m, 4H)；Conformer 2: δ 9.06 (brs, 1H), 7.94 (s, 1H), 7.72 (brs, 1H), 7.60 7.52 (m, 2H), 4.60 (s,2H),4.01(brs,2H),2.98(brs,2H),2.10(m,1H),0.77(m,4H)；¹³C NMR(125MHz, DMSO-d₆) conformer 1: δ 172.17,165.39,158.74 (d, J=236.9Hz, C-F), 149.18,138.31, (137.84,136.01,134.55,134.17,130.10,126.66 d, J=25.2Hz, CH-CF), 123.41,110.54 (d, J= 26.0Hz,CH-CF),102.01,45.21,43.07,26.17,11.34,7.86；Conformer 2: δ 172.17,165.39, 158.74 (d, J=236.9Hz, C-F), 149.18,138.31,137.84,136.01,134.55,134.17,130.10,126.66 (d, J=25.2Hz, CH-CF), 123.41,110.54 (d, J=26.0Hz, CH-CF), 102.01,49.05,42.75,25.01, 11.06,7.49；HRMS(ESI):m/z Calcd.For C₁₉H₁₈FN₄O₂S[M+H]⁺:385.1134；Found: 385.1125。

The fluoro-2-of embodiment 12. 6-(5-(2-propoxyl group acetyl group)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzene And [d] imidazoles-4-Methanamide, the preparation of I-12:

The fluoro-2-of step 1. 6-(5-(2-propoxyl group acetyl group)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] Imidazoles-4-methyl formate:

According to preparing the operational approach of compound I-11, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-5) (182mg, 0.60mmol) and 2-propoxyl group chloroacetic chloride (98mg, 0.72mmol) Preparing white powder 228mg for raw material, productivity is 88%.

¹H NMR(300MHz,DMSO-d₆) δ 8.09 (s, 1H), 7.77 (d, J=8.7Hz, 1H), 7.56 (d, J=9.3 Hz, 1H), 4.60 (s, 2H), 4.23 (s, 2H), 3.99 (s, 3H), 3.78 (t, J=5.1Hz, 2H), 3.41 (t, J=5.1Hz, 2H),2.90(brs,2H),1.60–1.46(m,2H),0.92–0.82(m,3H)；MS(ESI)432[M+H]⁺。

The fluoro-2-of step 2. 6-(5-(2-propoxyl group acetyl group)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] Imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-, (5-(2-propoxyl group acetyl group)-4,5,6,7-Tetramethylene sulfides are also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate (5-8) (228mg, 0.53mmol) is raw material ammonolysis Obtaining light tan solid 207mg, productivity is 94% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ8.96(brs,1H),7.90(s,1H),7.77–7.43(m,3H),4.57 (s, 2H), 4.21 (s, 2H), 3.85 3.69 (m, 2H), 3.39 (s, 2H), 3.05 2.75 (m, 2H), 1.51 (q, J=6.2Hz, 2H), 0.85 (t, J=6.5Hz, 3H)；¹³C NMR(125MHz,DMSO-d₆) conformer 1: δ 168.55, 165.64,158.67 (d, J=236.9Hz, C-F), 149.38,138.57,137.54,136.29,133.99,130.21,126.70 (d, J=27.7Hz, CH-CF), 110.29 (d, J=25.2Hz, CH-CF), 102.68,72.65,70.10,44.51,42.49, 25.85,22.85,10.99；Conformer 2: δ 8.96 (brs, 1H), 7.90 (s, 1H), 7.77 7.43 (m, 3H), 4.57 (s, 2H), 4.21 (s, 2H), 3.85 3.69 (m, 2H), 3.39 (s, 2H), 3.05 2.75 (m, 2H), 1.51 (q, J=6.2Hz, 2 H), 0.85 (t, J=6.5Hz, 3H)；HRMS(ESI):m/z Calcd.For C₂₀H₂₂FN₄O₃S[M+H]⁺:417.1397； Found:417.1398。

Embodiment 13. 2-(5-benzoyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles -4-Methanamide, the preparation of I-13:

Step 1. 2-(5-benzoyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methyl formate:

According to preparing the operational approach of compound I-11, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-5) (182mg, 0.60mmol) and Benzenecarbonyl chloride. (101mg, 0.72mmol) Preparing white powder 238mg for raw material, productivity is 91%.

¹H NMR (400MHz, Chloroform-d) conformer 1: δ 10.5 (brs, 1H), 8.12 (d, J=8.1Hz, 1H),7.69–7.57(m,2H),7.50–7.43(m,5H),4.87(s,2H),4.01(s,3H),3.75(brs,2H),2.97 (brs,2H)；Conformer 2: δ 10.5 (brs, 1H), 8.12 (d, J=8.1Hz, 1H), 7.69 7.57 (m, 2H), 7.50–7.43(m,5H),4.57(s,2H),4.12(brs,2H),4.01(s,3H),3.06(brs,2H)；MS(ESI)436 [M+H]⁺。

Step 2. 2-(5-benzoyl-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methanamide:

According to preparing the operational approach of compound I-1, with 2-(5-benzoyl-4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2- Base)-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (5-9) (238mg, 0.55mmol) is that raw material ammonia solution obtains white Solid 159mg, productivity is 69% (dichloromethane: methanol=10:1).

¹H NMR(400MHz,DMSO-d₆) conformer 1: δ 9.07 (brs, 1H), 7.96 (s, 1H), 7.75 (brs, 1H),7.60–7.52(m,2H),7.52–7.43(m,5H),4.75(s,2H),3.64(brs,2H),2.95(brs,2H)；Structure As isomer 2: δ 9.07 (brs, 1H), 7.96 (s, 1H), 7.75 (brs, 1H), 7.60 7.52 (m, 2H), 7.52 7.43 (m, 5 H),4.55(s,2H),3.98(brs,2H),2.95(brs,2H)；¹³C NMR(125MHz,DMSO-d₆)δ170.42, (165.32,158.82 d, J=238.0Hz, C-F), 149.01,138.47,137.70,136.46,135.96,133.85,130.24, (129.02,127.22,126.75,123.38,110.52 d, J=23.0Hz, CH-CF), 101.88 (d, J=23.0Hz, CH-CF),45.12,42.69,25.83；HRMS(ESI):m/z Calcd.For C₂₂H₁₈FN₄O₂S[M+H]⁺:421.1134； Found:421.1128。

The fluoro-2-of embodiment 14. 6-(5-(mesyl)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] miaow Azoles-4-Methanamide, the preparation of I-14:

The fluoro-2-of step 1. 6-(5-(mesyl)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles -4-methyl formate:

According to preparing the operational approach of compound I-11, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [3,2-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-5) (182mg, 0.60mmol) and mesyl chloride (82mg, 0.72mmol) Preparing white powder 135mg for raw material, productivity is 55%.

¹H NMR(400MHz,DMSO-d₆) δ 12.69 (brs, 1H), 8.08 (s, 1H), 7.78 (dd, J=9.0,2.0Hz, 1H), 7.56 (dd, J=10.0,2.3Hz, 1H), 4.36 (s, 2H), 3.99 (s, 3H), 3.54 (t, J=5.6Hz, 2H), 3.00 (brs,5H)；MS(ESI)436[M+H]⁺。

The fluoro-2-of step 2. 6-(5-(mesyl)-4,5,6,7-Tetramethylene sulfide also [3,2-c] pyridine-2-base)-1H-benzo [d] imidazoles -4-Methanamide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(5-(mesyl)-4,5,6,7-Tetramethylene sulfides also [3,2-c] Pyridine-2-base) to be that raw material ammonia solution obtains white for-1H-benzo [d] imidazoles-4-methyl formate (5-10) (135mg, 0.33mmol) Color powder 105mg, productivity is 81% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆)δ8.98(brs,1H),7.92(s,1H),7.73(brs,1H),7.62–7.48 (m, 2H), 4.36 (s, 2H), 3.53 (t, J=5.6Hz, 2H), 2.99 (brs, 5H)；¹³C NMR(125MHz, DMSO-d₆) δ 165.45,158.73 (d, J=236.0Hz, C-F), 149.15,144.29,137.33,133.00,132.99, 130.30,126.58,123.27 (d, J=24.2Hz, CH-CF), 110.44 (d, J=25.2Hz, CH-CF), 102.10,45.39, 43.35,35.96,25.37；HRMS(ESI):m/z Calcd.For C₁₆H₁₆FN₄O₃S₂[M]⁺:395.0648；Found: 395.0639。

The fluoro-2-of embodiment 15. 6-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, The preparation of I-15:

Step 1. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid The tert-butyl ester (2-2) (726mg, 2.72mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (500mg, 2.72mmol) be that raw material prepares yellow foamy solid 973mg, productivity be 83% (petroleum ether: ethyl acetate= 3:1).

¹H NMR(300MHz,DMSO-d₆) δ 12.60 (brs, 1H), 8.05 (s, 1H), 7.75 (dd, J=9.0,2.1Hz, 1H), 7.53 (dd, J=9.5,2.1Hz, 1H), 4.61 (s, 2H), 3.96 (s, 3H), 3.62 (t, J=5.7Hz, 2H), 2.69 (t, J=5.7Hz, 2H), 1.42 (s, 9H)；MS(ESI)432[M+H]⁺。

The fluoro-2-of step 2. 6-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7- Dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-6) (500mg, 1.16mmol) is that raw material deprotects base Obtaining pale yellow foam solid 303mg, productivity is 79% (dichloromethane: methanol=50:1).

¹H NMR(300MHz,DMSO-d₆) δ 8.00 (brs, 1H), 7.72 (dd, J=9.4,2.1Hz, 1H), 7.52 (dd, J=9.9,2.1Hz, 1H), 3.96 (s, 3H), 3.90 (s, 2H), 2.94 (t, J=5.7Hz, 2H), 2.59 (t, J=5.6Hz, 2 H)；MS(ESI)332[M+H]⁺。

The fluoro-2-of step 3. 6-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-6) (100mg, 0.30mmol) is that raw material ammonia solution obtains white powder 72mg, Productivity is 76% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ9.05(brs,1H),7.94(s,1H),7.72(brs,1H),7.61–7.54 (m,2H),3.96(s,2H),2.99(brs,2H),2.65(brs,2H)；¹³C NMR(125MHz,DMSO-d₆)δ (165.62,158.60 d, J=234.0Hz, C-F), 149.79,139.68,135.98,129.11,128.95 (d, J=26.0Hz, CH-CF), 110.12 (d, J=25.8Hz, CH-CF), 44.77,42.92,26.47；HRMS(ESI):m/z Calcd.For C₁₅H₁₄FN₄OS[M+H]⁺:317.0872；Found:317.0861.

The fluoro-2-of embodiment 16. 6-(6-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-16:

The fluoro-2-of step 1. 6-(6-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 5-1, with the fluoro-2-of 6-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-6) (100mg, 0.30mmol) is raw material, through the light yellow solid that methylates to obtain 77mg, productivity is 74% (dichloromethane: methanol=30:1).

¹H NMR(300MHz,CD₃OD) δ 7.68 (s, 1H), 7.54 (dd, J=9.9,2.5Hz, 1H), 7.48 (dd, J= 8.7,2.5Hz,1H),4.01(s,3H),3.76(s,2H),2.86(brs,4H),2.54(s,3H)；MS(ESI)346 [M+H]⁺。

The fluoro-2-of step 2. 6-(6-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(6-methyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (5-11) (77mg, 0.22mmol) is that raw material ammonia solution obtains canescence powder End 36mg, productivity is 55% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ9.08(brs,1H),7.94(s,1H),7.68(brs,1H),7.59–7.52 (m, 2H), 3.60 (s, 2H), 2.73 (t, J=5.3Hz, 2H), 2.67 (t, J=5.6Hz, 2H), 2.39 (s, 3H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.49,158.72 (d, J=235.6Hz, C-F), 149.43,137.74,136.05, (135.16,129.23,128.59,122.98,110.32 d, J=26.5Hz, CH-CF), 101.83,53.72,51.99,45.40, 25.65；HRMS(ESI):m/z Calcd.For C₁₆H₁₆FN₄OS[M+H]⁺:331.1029；Found:331.1020.

The fluoro-2-of embodiment 17. 6-(7-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-17:

Step 1. N-(2-(thiene-3-yl) ethyl) acetamide:

At 0 DEG C, to dissolved with 2-(thiene-3-yl) ethylamine hydrochloride (15) (492mg, 3.0mmol) and triethylamine (9.0 Mmol) dichloromethane solution is slowly added dropwise chloroacetic chloride (283mg, 3.6mmol), is stirred at room temperature 30 minutes.Instead Answering liquid dichloromethane to extract (50mL × 3), organic layer washes with water successively, and saturated common salt is washed, and retains organic layer, uses Anhydrous sodium sulfate is dried, and filters concentrating under reduced pressure give light yellow oil 497mg, and productivity is 98%, is directly entered next step Reaction.

¹H NMR (300MHz, Chloroform-d) δ 7.32 7.32 (m, 1H), 7.01 (s, 1H), 6.95 (d, J=4.6 Hz, 1H), 5.02 (brs, 1H), 3.55 3.46 (m, 2H), 2.85 (t, J=6.9Hz, 2H), 1.95 (s, 3H)；MS(ESI) 170[M+H]⁺。

Step 2. 7-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

N-(2-(thiene-3-yl) ethyl) acetamide (16-1) (497mg, 2.9mmol) is dissolved in 3mL phosphorus oxychloride, It is heated to 60 DEG C, reacts two hours, cooling, reactant liquor is poured in frozen water, uses saturated Na₂CO₃Solution is neutralized to Neutrality, is extracted with ethyl acetate (50mL × 3), and organic layer washes with water successively, and saturated common salt is washed, and retains organic layer, It is dried with anhydrous sodium sulfate, filters and be concentrated under reduced pressure to give yellow oil, this grease is dissolved in 5mL absolute methanol, At 0 DEG C, add NaBH₄(220mg, 5.8mmol), is stirred at room temperature 1 hour, and reactant liquor is extracted with ethyl acetate (50mL × 3), organic layer washes with water successively, and saturated common salt is washed, and retains organic layer, is dried with anhydrous sodium sulfate, mistake Filter is concentrated under reduced pressure to give yellow oil, is dissolved in 5mL dichloromethane by this grease, at 0 DEG C, adds two carbonic acid two The tert-butyl ester (632mg, 2.9mmol), is stirred at room temperature 30 minutes, and concentrating under reduced pressure obtains thick product, through column chromatography (oil Ether: ethyl acetate=30:1) obtain colorless oil 506mg, three step gross production rates are 69%.

¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.0Hz, 1H), 6.75 (d, J=5.1Hz, 1H), 5.28 (s, 1H), 4.30 (s, 1H), 3.03 (t, J=12.6Hz, 1H), 2.77 2.54 (m, 2H), 1.48 (s, 9H), 1.46 (d, J=6.7Hz, 3H)；MS(ESI)276[M+Na]⁺。

Step 3. 2-bromine-7-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 10-1, with 7-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (14-3) (506mg, 2.0mmol) is that raw material prepares 465mg grease through bromo, productivity be 70% (petroleum ether: Ethyl acetate=30:1).

¹H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.16(brs,1H),4.25(brs,1H),3.00(t, J=11.0Hz, 1H), 2.73 2.43 (m, 2H), 1.48 (s, 9H), 1.41 (d, J=6.7Hz, 3H)；MS(ESI)354 [M+Na]⁺。

Step 4. 2-formoxyl-7-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-1, with 2-bromine-7-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-3) (465mg, 1.4mmol) is raw material, with tert-butyl lithium hexane solution (1.6M) generation For n-BuLi tetrahydrofuran solution, preparing pale yellow oil 295mg, productivity is 75% (petroleum ether: acetic acid Ethyl ester=8:1).

¹H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.33(brs,1H),4.33(brs, 1H), 3.02 (t, J=12.8Hz, 1H), 2.70 (m, 2H), 1.49 (d, J=6.7Hz, 3H), 1.49 (s, 9H)；MS(ESI) 304[M+Na]⁺。

Step 5. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7-methyl-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-7-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-3) (295mg, 1.05mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (193mg, 1.05mmol) is that raw material prepares pale yellow powder 262mg, and productivity is 59% (petroleum ether: acetic acid Ethyl ester=3:1).

¹H NMR (300MHz, Chloroform-d) δ 7.63 (dd, J=8.9,2.5Hz, 1H), 7.59 (dd, J=9.5,2.5 Hz, 1H), 7.35 (s, 1H), 5.34 (brs, 1H), 4.34 (brs, 1H), 4.02 (s, 3H), 3.06 (t, J=11.2Hz, 1H), 2.86 2.54 (m, 1H), 1.51 (d, J=6.8Hz, 3H), 1.51 (s, 9H)；MS(ESI)446[M+H]⁺。

The fluoro-2-of step 6. 6-(7-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7- Methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-7) (262mg, 0.59mmol) is raw material Deprotection base obtains white powder 123mg, and productivity is 60% (dichloromethane: methanol=50:1).

¹H NMR(300MHz,Methanol-d₄) δ 7.64 (s, 1H), 7.53 (dd, J=9.9,2.4Hz, 1H), 7.47 (dd, J=8.8,2.5Hz, 1H), 4.14 (q, J=6.9Hz, 1H), 4.00 (s, 3H), 3.32 3.28 (m, 1H), 2.97 (ddd, J= 12.8,9.8,5.1Hz, 1H), 2.86 2.60 (m, 2H), 1.49 (d, J=6.7Hz, 3H)；MS(ESI)346[M+H]⁺。

The fluoro-2-of step 7. 6-(7-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the fluoro-2-of methyl 6-(7-methyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyrrole Pyridine-2-base) to be that raw material ammonia solution obtains yellowish for-1H-benzo [d] imidazoles-4-methyl formate (4-7) (123mg, 0.35mmol) Color powder 102mg, productivity is 87% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆)δ8.96(brs,1H),7.90(brs,1H),7.68(s,1H),7.54(dd,J =6.3,3.0Hz, 1H), 7.51 (dd, J=4.4,3.0Hz, 1H), 4.17 (q, J=6.6Hz, 2H), 3.28-3.17 (m, 1H), 2.99-2.86 (m, 1H), 2.73-2.58 (m, 2H), 1.41 (d, J=6.6Hz, 3H)；¹³C NMR(125MHz, DMSO-d₆) δ 165.88,158.92 (d, J=237.4Hz, C-F), 149.34,143.87,135.76,129.20,129.02, 122.86,110.32 (d, J=26.9Hz, CH-CF), 102.04 (d, J=25.6Hz, CH-CF), 50.37,41.89,25.74, 22.74；HRMS(ESI):m/z Calcd.For C₁₆H₁₆FN₄OS[M+H]⁺:331.1029；Found:331.1023.

Embodiment 18. 2-(7-ethyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-18:

Step 1. N-(2-(thiene-3-yl) ethyl) propionic acid amide.:

According to preparing the operational approach of 16-1, with 2-(thiene-3-yl) ethylamine hydrochloride (492mg, 3.0mmol), Propionyl chloride (333mg, 3.6mmol) is that raw material prepares light yellow oil 543mg, and productivity is 99%, is directly entered Next step reaction.

¹H NMR (300MHz, Chloroform-d) δ 7.29 (dd, J=5.0,2.9Hz, 1H), 7.02 6.99 (m, 1H), 6.95 (dd, J=4.9,1.3Hz, 1H), 5.46 (brs, 1H), 3.52 (dd, J=12.8,6.8Hz, 2H), 2.85 (t, J=6.9 Hz, 2H), 2.17 (q, J=7.6Hz, 2H), 1.13 (t, J=7.6Hz, 3H)；MS(ESI)184[M+H]⁺。

Step 2. 7-ethyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of 14-3, with N-(2-(thiene-3-yl) ethyl) propionic acid amide. (16-2) (543mg, 3.0 Mmol) being that raw material prepares colorless oil 529mg, three step gross production rates are 66% (petroleum ether: ethyl acetate=30:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.0Hz, 1H), 6.76 (d, J= 5.0Hz,1H),5.22(brs,1H),4.28–4.07(m,1H),3.15–2.89(m,2H),2.57–2.51(m,1H), 1.90 1.74 (m, 2H), 1.47 (s, 9H), 1.03 (t, J=7.4Hz, 3H)；Conformer 2:¹H NMR(300MHz, Chloroform-d) δ 7.13 (d, J=5.0Hz, 1H), 6.76 (d, J=5.0Hz, 1H), 5.10 (brs, 1H), 4.49 4.28 (m, 1H), 2.86 2.64 (m, 2H), 2.62 2.58 (m, 1H), 1.90 1.74 (m, 2H), 1.47 (s, 9H), 1.03 (t, J= 7.4Hz,3H)；MS(ESI)290[M+Na]⁺。

Step 3. 2-bromo-7-ethyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 7-ethyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (14-4) (529mg, 2.0mmol) is that raw material prepares 408mg white solid through bromo, and productivity is 60% (oil Ether: ethyl acetate=30:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ6.72(s,1H),5.06(brs,1H), 4.23 4.00 (m, 1H), 3.19 2.86 (m, 2H), 2.45 (d, J=3.9Hz, 1H), 1.77 (p, J=7.3Hz, 2H), (1.47 s, 9H), 1.00 (t, J=7.4Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ 6.72(s,1H),4.97(brs,1H),4.46–4.23(m,1H),4.23–4.00(m,1H),2.80–2.55(m,2H), (2.51 d, J=4.1Hz, 1H), 1.77 (p, J=7.3Hz, 2H), 1.47 (s, 9H), 1.00 (t, J=7.4Hz, 3H)；MS (ESI)368[M+Na]⁺。

Step 4. 7-ethyl-2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with 2-bromo-7-ethyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-4) (408mg, 1.2mmol) is that raw material prepares pale yellow oil 329mg, and productivity is 93% (petroleum ether: ethyl acetate=8:1).

¹H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.45(s,1H),5.40–5.01(m,1H), 4.60 4.10 (m, 1H), 3.17 2.51 (m, 3H), 1.96 1.75 (m, 2H), 1.48 (d, J=1.6Hz, 9H), 1.04 (t, J =7.4Hz, 3H)；MS(ESI)318[M+Na]⁺。

Step 5. 7-ethyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 7-ethyl-2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-4) (329mg, 1.12mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (206mg, 1.12mmol) is that raw material prepares pale yellow powder 329mg, and productivity is 64% (petroleum ether: acetic acid Ethyl ester=3:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 10.41 (s, 1H), 7.63 (dd, J=8.9,2.5 Hz, 1H), 7.59 (dd, J=9.7,2.4Hz, 1H), 7.36 (s, 1H), 5.30 (brs, 1H), 4.33 4.20 (m, 1H), 4.01 (s,3H),3.13–2.94(m,2H),2.64–2.55(m,1H),1.94–1.78(m,2H),1.74–1.56(m,2H),1.07 (t, J=7.4Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ10.41(s,1H),7.63 (dd, J=8.9,2.5Hz, 1H), 7.59 (dd, J=9.7,2.4Hz, 1H), 7.36 (s, 1H), 5.16 (brs, 1H), 4.52–4.36(m,1H),4.01(s,3H),2.86–2.71(m,2H),2.68–2.64(m,1H),1.94–1.78(m,2H), 1.74 1.56 (m, 2H), 1.07 (t, J=7.4Hz, 3H)；MS(ESI)460[M+H]⁺。

Step 6. 2-(7-ethyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 7-ethyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles -2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-8) (329mg, 0.72mmol) is raw material Deprotection base obtains white powder 144mg, and productivity is 56% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.40 (brs, 1H), 7.62 (dd, J=9.2,1.3Hz, 1H), 7.59 (dd, J=9.5,1.3Hz, 1H), 7.37 (s, 1H), 4.01 (s, 3H), 3.36 (ddd, J=12.7,5.3,3.3Hz, 1H), 3.09 2.93 (m, 1H), 2.82 2.59 (m, 2H), 2.07 1.85 (m, 2H), 1.10 (t, J=7.3Hz, 3H)；MS(ESI) 360[M+H]⁺。

Step 7. 2-(7-ethyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with 2-(7-ethyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2-base)-6- Fluoro-1H-benzo [d] imidazoles-4-methyl formate (4-8) (144mg, 0.40mmol) is that raw material ammonia solution obtains pale yellow powder 119mg, productivity is 86% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ9.00(brs,1H),7.97(brs,1H),7.78(s,1H),7.56(dd,J =10.7,2.7Hz, 1H), 7.53 (dd, J=8.3,2.7Hz, 1H), 4.23 (t, J=6.0Hz, 1H), 3.34 (dt, J=12.4, 4.1Hz, 1H), 3.06 (dt, J=12.4,4.1Hz, 1H), 2.87 2.71 (m, 2H), 1.99-1.90 (m, 1H), 1.89 1.76 (m, 1H), 1.08 (t, J=7.3Hz, 3H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.56,158.71 (d, J= 238.1Hz, C-F), 149.09,140.19,135.63,130.15,128.89,122.95,110.39 (d, J=26.3Hz, CH-CF), 102.48 (d, J=27.1Hz, CH-CF), 55.63,41.39,28.94,24.58,10.69；HRMS(ESI):m/z Calcd.For C₁₇H₁₈FN₄OS[M]⁺:345.1185；Found:345.1178.

The fluoro-2-of embodiment 19. 6-(7-propyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-19:

Step 1. N-(2-(thiene-3-yl) ethyl) butyramide:

According to preparing the operational approach of 16-1, with 2-(thiene-3-yl) ethylamine hydrochloride (492mg, 3.0mmol), Butyl chloride (383mg, 3.6mmol) is that raw material prepares light yellow oil 585mg, and productivity is 99%, is directly entered Next step reaction.

¹H NMR (300MHz, Chloroform-d) δ 7.32 7.28 (m, 1H), 7.02 6.98 (m, 1H), 6.95 (d, J= 4.8Hz, 1H), 5.43 (brs, 1H), 3.57 3.48 (m, 2H), 2.85 (t, J=6.8Hz, 2H), 2.11 (dd, J=8.3,6.7 Hz, 2H), 1.70 1.60 (m, 2H), 0.92 (t, J=7.4Hz, 3H)；MS(ESI)198[M+H]⁺。

Step 2. 7-propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of 14-3, with N-(2-(thiene-3-yl) ethyl) butyramide (16-3) (585mg, 3.0 Mmol) being that raw material prepares colorless oil 532mg, three step gross production rates are 63% (petroleum ether: ethyl acetate=30:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.1Hz, 1H), 6.76 (d, J= 5.1Hz, 1H), 5.29 (brs, 1H), 4.25 4.09 (m, 1H), 3.18 2.91 (m, 2H), 2.54 (d, J=4.2Hz, 1H), 1.84 1.64 (m, 2H), 1.55 1.42 (m, 2H), 1.47 (s, 9H), 0.96 (t, J=7.3Hz, 3H)；Conformer 2:¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.1Hz, 1H), 6.76 (d, J=5.1Hz, 1H), 5.17 (brs, 1H), 4.44 4.30 (m, 1H), 2.86 2.65 (m, 2H), 2.59 (d, J=3.9Hz, 1H), 1.84 1.64 (m, 2 H), 1.55 1.42 (m, 2H), 1.47 (s, 9H), 0.96 (t, J=7.3Hz, 3H)；MS(ESI)304[M+Na]⁺。

Step 3. 2-bromo-7-propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 7-propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (14-5) (532mg, 1.9mmol) is that raw material prepares 512mg clear crystal through bromo, and productivity is 76% (oil Ether: ethyl acetate=30:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.24–5.10(m,1H), 4.21–4.03(m,1H),3.18–2.84(m,2H),2.49–2.37(m,1H),1.85–1.61(m,2H),1.57–1.39(m, 2H), 1.47 (s, 9H), 0.95 (t, J=7.3Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d) δ6.71(s,1H),5.11–4.95(m,1H),4.42–4.26(m,1H),2.84–2.56(m,2H),2.54–2.47(m,1H), 1.85 1.61 (m, 2H), 1.57 1.39 (m, 2H), 1.47 (s, 9H), 0.95 (t, J=7.3Hz, 3H)；MS(ESI)382 [M+Na]⁺。

Step 4. 2-formoxyl-7-propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with 2-bromo-7-propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-5) (512mg, 1.4mmol) is that raw material prepares pale yellow oil 426mg, and productivity is 96% (petroleum ether: ethyl acetate=8:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.49–5.29 (m,1H),4.30–4.16(m,1H),3.18–2.90(m,2H),2.63–2.52(m,1H),1.91–1.64(m,2H),1.48 (s, 9H), 0.97 (t, J=7.3Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ9.83(s, 1H),7.44(s,1H),5.29–5.10(m,1H),4.50–4.34(m,1H),2.87–2.69(m,2H),2.68–2.61(m, 1H), 1.91 1.64 (m, 2H), 1.48 (s, 9H), 0.97 (t, J=7.3Hz, 3H)；MS(ESI)332[M+Na]⁺。

Step 5. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7-propyl group-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-7-propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-5) (426mg, 1.38mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (254mg, 1.38mmol) is that raw material prepares pale yellow powder 562mg, and productivity is 86% (petroleum ether: acetic acid Ethyl ester=3:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 10.42 (s, 1H), 7.63 (dd, J=9.3,2.4 Hz, 1H), 7.58 (dd, J=9.4,2.4Hz, 1H), 7.35 (s, 1H), 5.36 (brs, 1H), 4.31 4.16 (m, 1H), 4.01 (s,3H),2.88–2.68(m,2H),2.64–2.55(m,1H),1.90–1.72(m,4H),1.64–1.39(m,2H),1.49 (s, 9H), 0.98 (t, J=7.3Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ10.42 (s, 1H), 7.63 (dd, J=9.3,2.4Hz, 1H), 7.58 (dd, J=9.4,2.4Hz, 1H), 7.35 (s, 1H), 5.21 (brs, 1H),4.50–4.34(m,1H),3.21–2.93(m,2H),2.69–2.61(m,1H),1.90–1.72(m,4H), 1.64 1.39 (m, 2H), 1.49 (s, 9H), 0.98 (t, J=7.3Hz, 3H)；MS(ESI)474[M+H]⁺。

The fluoro-2-of step 6. 6-(7-propyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7- Propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-9) (329mg, 0.72mmol) is raw material Deprotection base obtains faint yellow solid 237mg, and productivity is 88% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.39 (brs, 1H), 7.62 (dd, J=8.8,2.4Hz, 1H), 7.58 (dd, J=9.7,2.4Hz, 1H), 7.38 (s, 1H), 4.12 4.03 (m, 1H), 4.02 (s, 3H), 3.40 3.30 (m, 1H), 3.09 2.93 (m, 1H), 2.79 2.60 (m, 2H), 1.95 1.69 (m, 2H), 1.63 1.47 (m, 2H), 1.00 (t, J= 7.3Hz,3H)；MS(ESI)374[M+H]⁺。

The fluoro-2-of step 7. 6-(7-propyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(7-propyl group-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (4-9) (237mg, 0.63mmol) is that raw material ammonia solution obtains white powder 137mg, productivity is 61% (dichloromethane: methanol=10:1).

¹H NMR(400MHz,DMSO-d₆)δ8.99(brs,1H),7.93(brs,1H),7.69(s,1H),7.54(dd,J =10.9,2.5Hz, 1H), 7.51 (dd, J=8.6,2.6Hz, 1H), 4.12 (dd, J=8.1,4.6Hz, 1H), 3.23 (dt, J =12.6,4.8Hz, 1H), 2.94 (ddd, J=13.2,8.9,5.4Hz, 1H), 2.78 2.57 (m, 2H), 1.84-1.62 (m, 2 H), 1.59 1.41 (m, 2H), 0.93 (t, J=7.3Hz, 3H)；¹³C NMR(100MHz,DMSO-d₆)δ165.47, 158.64 (d, J=236.1Hz, C-F), 149.21,142.22,135.84,129.42,128.92,122.91,110.29 (d, J= 25.9Hz, CH-CF), 102.02 (d, J=26.3Hz, CH-CF), 54.22,41.62,38.79,25.44,19.09,14.31； HRMS(ESI):m/z Calcd.For C₁₈H₂₀FN₄OS[M+H]⁺:359.1342；Found:359.1334.

Embodiment 20. 2-(7-butyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-20:

Step 1. N-(2-(thiene-3-yl) ethyl) pentanamide:

According to preparing the operational approach of 16-1, with 2-(thiene-3-yl) ethylamine hydrochloride (492mg, 3.0mmol), Valeric chloride (434mg, 3.6mmol) is that raw material prepares light yellow oil 627mg, and productivity is 99%, is directly entered Next step reaction.

¹H NMR(300MHz,Chloroform-d)δ7.32–7.27(m,1H),7.02–6.99(m,1H),6.98–6.94 (m,1H),5.46(brs,1H),3.56–3.48(m,1H),3.03–2.93(m,1H),2.88–2.76(m,1H), 2.16 2.09 (m, 1H), 1.59 1.52 (m, 2H), 1.37 1.28 (m, 2H), 0.90 (t, J=7.3Hz, 3H)；MS(ESI) 212[M+H]⁺。

Step 2. 7-butyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of 14-3, with N-(2-(thiene-3-yl) ethyl) pentanamide (16-4) (627mg, 3.0 Mmol) be that raw material prepares pale yellow oil 354mg, three step gross production rates be 40% (petroleum ether: ethyl acetate=30: 1)。

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.0Hz, 1H), 6.76 (d, J= 5.0Hz,1H),5.29(brs,1H),4.28–4.00(m,1H),3.16–2.87(m,2H),2.57–2.45(m,1H), 1.86 1.67 (m, 2H), 1.47 (s, 9H), 1.45 1.21 (m, 4H), 0.91 (t, J=6.8Hz, 3H)；Conformer 2:¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.0Hz, 1H), 6.76 (d, J=5.0Hz, 1H), 5.16 (brs,1H),4.48–4.28(m,1H),2.85–2.62(m,2H),2.62–2.57(m,1H),1.86–1.67(m,2H), 1.47 (s, 9H), 1.45 1.21 (m, 4H), 0.91 (t, J=6.8Hz, 3H)；MS(ESI)318[M+H]⁺。

Step 3. 2-bromo-7-butyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 7-butyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (14-6) (354mg, 1.2mmol) is that raw material prepares 347mg colorless oil through bromo, and productivity is 77% (stone Oil ether: ethyl acetate=30:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.22–5.12(m,1H), 4.21 4.04 (m, 1H), 3.10 2.89 (m, 2H), 2.45 (d, J=4.0Hz, 1H), 1.84 1.63 (m, 2H), 1.47 (s, 9H), 1.43 1.29 (m, 4H), 0.91 (t, J=6.9Hz, 3H)；Conformer 2:¹H NMR(300MHz, Chloroform-d)δ6.71(s,1H),5.09–4.94(m,1H),4.40–4.23(m,1H),2.75–2.56(m,2H), (2.50 d, J=3.3Hz, 1H), 1.84 1.63 (m, 2H), 1.47 (s, 9H), 1.43 1.29 (m, 4H), 0.91 (t, J=6.9 Hz,3H)；MS(ESI)396[M+Na]⁺。

Step 4. 7-butyl-2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with 2-bromo-7-butyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-6) (347mg, 0.9mmol) is that raw material prepares colorless oil 240mg, and productivity is 81% (petroleum ether: ethyl acetate=8:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.42–5.28 (m, 1H), 4.31 4.17 (m, 1H), 3.15 2.90 (m, 2H), 2.63 2.55 (m, 1H), 1.79 (dd, J=6.5,5.1Hz, 2H), 1.48 (s, 9H), 1.45 1.24 (m, 4H), 0.92 (t, J=6.8Hz, 3H)；Conformer 2:¹H NMR(300 MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.28–5.09(m,1H),4.50–4.33(m,1H), 2.84 2.68 (m, 2H), 2.69 2.61 (m, 1H), 1.79 (dd, J=6.5,5.1Hz, 2H), 1.48 (s, 9H), 1.45 1.24 (m, 4H), 0.92 (t, J=6.8Hz, 3H)；MS(ESI)346[M+Na]⁺。

Step 5. 7-butyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-7-propyl group-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-5) (426mg, 1.38mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (254mg, 1.38mmol) is that raw material prepares pale yellow powder 524mg, and productivity is 78% (petroleum ether: acetic acid Ethyl ester=3:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.64 (dd, J=8.9, 2.5Hz, 1H), 7.61 (dd, J=9.8,2.5Hz, 1H), 7.35 (s, 1H), 5.43 5.27 (m, 1H), 4.32 4.17 (m, 1 H), 4.02 (s, 3H), 3.21 2.92 (m, 2H), 2.64 2.55 (m, 1H), 1.83 (q, J=7.3Hz, 2H), 1.50 (s, 9 H), 1.47 1.20 (m, 4H), 0.93 (t, J=7.0Hz, 3H)；Conformer 2:¹H NMR(300MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.64 (dd, J=8.9,2.5Hz, 1H), 7.61 (dd, J=9.8,2.5Hz, 1H), 7.35(s,1H),5.26–5.13(m,1H),4.50–4.33(m,1H),4.02(s,3H),2.87–2.70(m,2H), 2.70 2.63 (m, 1H), 1.83 (q, J=7.3Hz, 2H), 1.50 (s, 9H), 1.47 1.20 (m, 4H), 0.93 (t, J=7.0 Hz,3H)；MS(ESI)488[M+H]⁺。

The fluoro-2-of step 6. 6-(7-butyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 7-butyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles -2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (4-10) (524mg, 1.07mmol) is former Material deprotection base obtains white powder 240mg, and productivity is 58% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.40 (brs, 1H), 7.62 (dd, J=9.0,2.5Hz, 1H), 7.58 (dd, J=9.6,2.3Hz, 1H), 7.38 (s, 1H), 4.14 4.03 (m, 1H), 4.01 (s, 3H), 3.44 3.30 (m, 1H), 3.10–2.95(m,1H),2.82–2.62(m,1H),1.84–1.65(m,2H),1.61–1.43(m,1H),1.46–1.33(m, 2H), 0.94 (t, J=7.1Hz, 3H)；MS(ESI)388[M+H]⁺。

Step 7. 2-(7-butyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(7-butyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (4-10) (240mg, 0.62mmol) is that raw material ammonia solution obtains faint yellow powder End 178mg, productivity is 77% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ9.03(brs,1H),7.93(brs,1H),7.67(s,1H),7.55(dd,J =14.8,2.4,1H), 7.53 (dd, J=12.8,2.4,1H), 7.52 (d, J=2.4Hz, 1H), 3.98-3.89 (m, 1H), (3.15 dt, J=12.4,4.4Hz, 1H), 2.83 (ddd, J=13.1,8.8,5.0Hz, 1H), 2.67-2.54 (m, 2H), 1.82 1.71 (m, 1H), 1.69-1.60 (m, 1H), 1.52-1.41 (m, 2H), 1.41 1.27 (m, 2H), 0.90 (t, J=7.3 Hz,3H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.57,158.66 (d, J=237.0Hz, C-F), 149.55, (144.43,136.38,129.17,128.78,122.97,110.24 d, J=26.2Hz, CH-CF), 102.18 (d, J=27.9 Hz,CH-CF),54.61,42.05,37.09,28.12,26.55,22.66,14.45；HRMS(ESI):m/z Calcd.For C₁₉H₂₂FN₄OS[M+H]⁺:373.1498；Found:373.1490.

The fluoro-2-of embodiment 21. 6-(7-isopropyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles -4-Methanamide, the preparation of I-21:

Step 1. N-(2-(thiene-3-yl) ethyl) isobutyramide:

According to preparing the operational approach of 16-1, with 2-(thiene-3-yl) ethylamine hydrochloride (492mg, 3.0mmol), Isobutyryl chloride (383mg, 3.6mmol) is that raw material prepares light yellow oil 585mg, and productivity is 99%, directly enters Enter next step reaction.

¹H NMR (300MHz, Chloroform-d) δ 7.29 (dd, J=4.9,3.0Hz, 1H), 7.01 6.98 (m, 1H), (6.96 dd, J=4.9,1.3Hz, 1H), 5.44 (brs, 1H), 3.55 3.47 (m, 2H), 2.85 (t, J=6.9Hz, 2H), 2.29 (hept, J=6.8Hz, 1H), 1.11 (d, J=6.9Hz, 6H)；MS(ESI)198[M+H]⁺。

Step 2. 7-isopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of 14-3, with N-(2-(thiene-3-yl) ethyl) isobutyramide (16-5) (585mg, 3.0 Mmol) being that raw material prepares white solid 611mg, three step gross production rates are 72% (petroleum ether: ethyl acetate=30:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.14 (d, J=5.0Hz, 1H), 6.79 (d, J= 5.0Hz,1H),5.07–4.97(m,1H),4.28–4.13(m,1H),3.21–2.95(m,1H),2.82–2.64(m,2H), (2.06 dt, J=13.3,6.7Hz, 1H), 1.47 (s, 9H), 1.14 (d, J=6.7Hz, 3H), 0.97 (d, J=6.9Hz, 3H)； Conformer 2:¹H NMR (300MHz, Chloroform-d) δ 7.14 (d, J=5.0Hz, 1H), 6.79 (d, J=5.0 Hz,1H),4.94–4.85(m,1H),4.50–4.36(m,1H),3.21–2.95(m,1H),2.82–2.64(m,2H),2.06 (dt, J=13.3,6.7Hz, 1H), 1.47 (s, 9H), 1.14 (d, J=6.7Hz, 3H), 0.97 (d, J=6.9Hz, 3H)；MS (ESI)304[M+Na]⁺。

Step 3. 2-bromo-7-isopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 7-isopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 tertiary fourth of (5H)-carboxylic acid Ester (14-7) (611mg, 2.2mmol) is that raw material prepares 672mg white solid through bromo, and productivity is 85% (stone Oil ether: ethyl acetate=30:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ6.74(s,1H),4.96–4.84(m,1H), 4.50 4.31 (m, 1H), 2.79 2.57 (m, 2H), 2.10 1.89 (m, 2H), 1.46 (s, 9H), 1.11 (d, J=6.7Hz, 3H), 0.96 (d, J=6.9Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ6.74(s, 1H),4.81–4.71(m,1H),4.24–4.05(m,1H),3.26–2.90(m,2H),2.10–1.89(m,2H),1.46(s,9 H), 1.11 (d, J=6.7Hz, 3H), 0.96 (d, J=6.9Hz, 3H)；MS(ESI)382[M+Na]⁺。

Step 4. 2-formoxyl-7-isopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with 2-bromo-7-isopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-7) (672mg, 1.9mmol) is that raw material prepares colorless oil 240mg, and productivity is 40% (petroleum ether: ethyl acetate=8:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.47(s,1H),5.15–5.07 (m,1H),4.31–4.20(m,1H),3.19–2.94(m,2H),2.17–1.90(m,2H),1.48(s,9H),1.16(d,J =6.7Hz, 3H), 0.98 (d, J=6.9Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ 9.84(s,1H),7.47(s,1H),4.99–4.91(m,1H),4.54–4.43(m,1H),2.87–2.58(m,2H), 2.17 1.90 (m, 2H), 1.48 (s, 9H), 1.16 (d, J=6.7Hz, 4H), 1.16 (d, J=6.7Hz, 3H), 0.98 (d, J =6.9Hz, 3H)；MS(ESI)332[M+Na]⁺。

-7-isopropyl-4,7-dihydro-thiophene is also for step 5. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base) [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-7-isopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-7) (240mg, 0.78mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (143mg, 0.78mmol) is that raw material prepares yellow oil 333mg, and productivity is 90% (petroleum ether: acetic acid Ethyl ester=3:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ10.51(s,1H),7.74–7.47(m,2H), 7.38(s,1H),5.15–5.04(m,1H),4.33–4.21(m,1H),4.00(s,3H),3.21–2.99(m,1H), 2.89 2.54 (m, 2H), 2.25 2.06 (m, 1H), 1.18 (d, J=6.6Hz, 3H), 1.02 (d, J=6.8Hz, 3H)；Structure As isomer 2:¹H NMR(300MHz,Chloroform-d)δ10.51(s,1H),7.74–7.47(m,2H),7.38(s,1 H),5.00–4.90(m,1H),4.57–4.41(m,1H),4.00(s,3H),3.21–2.99(m,1H),2.89–2.54(m,2 H), 2.25 2.06 (m, 1H), 1.18 (d, J=6.6Hz, 3H), 1.02 (d, J=6.8Hz, 3H)；MS(ESI)475 [M+H]⁺。

The fluoro-2-of step 6. 6-(7-isopropyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Acid methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7- Isopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-11) (333mg, 0.89mmol) is former Material deprotection base obtains pale yellow powder 248mg, and productivity is 75% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.62 (dd, J=14.0,2.3Hz, 1H), 7.59 (dd, J=14.5,2.3Hz, 1H), 7.39 (s, 1H), 4.02 (s, 3H), 3.39 (ddd, J=13.0,5.3,2.7Hz, 1 H), 3.06 2.90 (m, 1H), 2.81 2.58 (m, 2H), 2.25 2.10 (m, 1H), 1.16 (d, J=7.0Hz, 3H), 0.95 (d, J=6.9Hz, 3H)；MS(ESI)374[M+H]⁺。

The fluoro-2-of step 7. 6-(7-isopropyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(7-isopropyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine -2-base)-1H-benzo [d] imidazoles-4-methyl formate (4-11) (248mg, 0.66mmol) is that raw material ammonia solution obtains light brown Powder 166mg, productivity is 70% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆) δ 9.08 (brs, 1H), 7.97 (brs, 1H), 7.76 (s, 1H), 7.55 (t, J= 10.8Hz,2H),4.24–4.17(m,1H),3.35–3.28(m,1H),3.03–2.95(m,1H),2.85-2.75(m,1H), 2.74 2.65 (m, 1H), 2.23-2.15 (m, 1H), 1.11 (d, J=6.9Hz, 3H), 0.95 (d, J=6.9Hz, 3H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.33,158.77 (d, J=236.9Hz), 149.04,138.46,136.85, (136.09,129.99,129.05,123.34,110.44 d, J=24.4Hz), 101.82 (d, J=26.9Hz), 59.80,42.35, 33.49,29.49,19.62,17.32.HRMS(ESI):m/z Calcd.For C₁₈H₂₀FN₄OS[M+H]⁺:359.1342； Found:359.1336。

Embodiment 22. 2-(7-cyclopropyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methanamide, the preparation of I-22:

Step 1. N-(2-(thiene-3-yl) ethyl) cyclopropyl carboxamide:

According to preparing the operational approach of 16-1, with 2-(thiene-3-yl) ethylamine hydrochloride (492mg, 3.0mmol), Cyclopropyl carbonyl chloride (376mg, 3.6mmol) is that raw material prepares light yellow oil 579mg, and productivity is 99%, directly Tap into and react into next step.

¹H NMR (300MHz, Chloroform-d) δ 7.32 7.27 (m, 1H), 7.02 (s, 1H), 6.96 (d, J=4.8 Hz, 1H), 5.64 (brs, 0H), 3.54 (dd, J=6.7,6.5Hz, 2H), 2.86 (t, J=6.7Hz, 2H), 1.32 1.21 (m, 1H),0.99–0.92(m,2H),0.75–0.66(m,2H)；MS(ESI)218[M+Na]⁺。

Step 2. 7-cyclopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of 14-3, with N-(2-(thiene-3-yl) ethyl) cyclopropyl carboxamide (16-6) (579mg, Being 3.0mmol) that raw material prepares white solid 373mg, three step gross production rates are 45% (petroleum ether: ethyl acetate=30:1).

¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.1Hz, 1H), 6.77 (d, J=5.1Hz, 1H), 4.62 4.25 (m, 1H), 3.26 3.06 (m, 1H), 2.81 2.52 (m, 2H), 1.47 (d, J=0.7Hz, 9H), 1.32–1.18(m,1H),0.83–0.61(m,2H),0.59–0.47(m,2H)；MS(ESI)302[M+Na]⁺。

Step 3. 2-bromo-7-cyclopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 7-cyclopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 tertiary fourth of (5H)-carboxylic acid Ester (14-8) (373mg, 1.4mmol) is that raw material prepares 382mg colorless oil through bromo, and productivity is 79% (stone Oil ether: ethyl acetate=30:1).

¹H NMR(300MHz,Chloroform-d)δ6.73(s,1H),4.42–4.29(m,2H),3.22–3.07(m,1 H),2.73–2.58(m,1H),2.57–2.44(m,1H),1.46(s,9H),1.31–1.17(m,1H),0.80–0.60(m,2 H),0.61–0.38(m,2H)；MS(ESI)380[M+Na]⁺。

Step 4. 7-cyclopropyl-2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with 2-bromo-7-cyclopropyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-8) (382mg, 1.1mmol) is that raw material prepares pale yellow oil 302mg, and productivity is 89% (petroleum ether: ethyl acetate=8:1).

¹H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.45(s,1H),4.53(brs,1H),4.40(brs, 1H),3.27–3.00(m,1H),2.87–2.58(m,2H),1.67–1.57(m,1H),1.47(s,9H),1.34–1.15(m, 1H),0.91–0.64(m,2H),0.65–0.49(m,2H)；MS(ESI)330[M+Na]⁺。

-4,7-dihydro-thiophene is also for step 5. 7-cyclopropyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base) [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 7-cyclopropyl-2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-8) (302mg, 0.98mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (181mg, 0.98mmol) is that raw material prepares white solid 438mg, and productivity is 95% (petroleum ether: acetic acid second Ester=3:1).

¹H NMR (300MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.63 (dd, J=8.9,2.4Hz, 1H), 7.58 (dd, J=9.6,2.4Hz, 1H), 7.36 (s, 1H), 4.53 (brs, 1H), 4.43 (brs, 2H), 4.01 (s, 3H), 3.29 3.10 (m,1H),2.86–2.58(m,2H),1.49(s,9H),0.92–0.67(m,2H),0.65–0.53(m,2H)；MS(ESI) 472[M+H]⁺。

Step 6. 2-(7-cyclopropyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Acid methyl ester:

According to preparing the operational approach of compound 4-1, with 7-cyclopropyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] miaow Azoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-12) (438mg, 0.80mmol) is Raw material deprotection base obtains white powder 298mg, and productivity is 86% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.41 (s, 1H), 7.63 (dd, J=8.9,2.3Hz, 1H), 7.57 (d, J=9.7Hz, 1H), 7.38 (s, 1H), 4.01 (s, 3H), 3.38 (ddd, J=12.4,5.6,2.7Hz, 1H), 3.14 (dd, J=9.5,2.2Hz, 1H), 2.99 (ddd, J=12.3,10.0,4.7Hz, 1H), 2.87 2.72 (m, 1H), 2.71 2.60 (m, 1H),1.20–1.05(m,1H),0.84–0.61(m,2H),0.60–0.41(m,2H)；MS(ESI)372[M+H]⁺。

Step 7. 2-(7-cyclopropyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Amide:

According to preparing the operational approach of compound I-1, with 2-(7-cyclopropyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base) to be that raw material ammonia solution obtains light brown for-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (4-12) (298mg, 0.60mmol) Color powder 260mg, productivity is 91% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆)δ9.00(brs,1H),7.88(brs,1H),7.68(s,1H),7.53(dd,J =11.0,2.6Hz, 1H), 7.49 (dd, J=9.6,2.6Hz, 1H), 3.28-3.17 (m, 2H), 2.92 2.76 (m, 1H), 2.76–2.54(m,2H),1.18–0.98(m,2H),0.75-0.62(m,1H),0.6–0.40(m,2H)；¹³C NMR(125 MHz,DMSO-d₆) δ 165.56,158.67 (d, J=236.6Hz, C-F), 149.54,143.14,135.92,129.37, 128.80,122.83,110.24 (d, J=26.2Hz, CH-CF), 102.16 (d, J=24.6Hz, CH-CF), 60.43,42.54, 29.50,25.92,18.06,4.30,3.84；HRMS(ESI):m/z Calcd.For C₁₈H₁₈FN₄OS[M+H]⁺:357.1185； Found:357.1179。

Embodiment 23. 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methanamide, the preparation of I-23:

Step 1. N-(2-(thiene-3-yl) ethyl) pivaloyl amine:

According to preparing the operational approach of 16-1, with 2-(thiene-3-yl) ethylamine hydrochloride (492mg, 3.0mmol), Pivaloyl chloride (434mg, 3.6mmol) is that raw material prepares light yellow oil 627mg, and productivity is 99%, directly enters Enter next step reaction.

¹H NMR(300MHz,Chloroform-d)δ7.31–7.27(m,1H),7.01–6.93(m,2H),5.6(brs,1 H), 3.53 3,45 (m, 2H), 2.85 (t, J=6.8Hz, 2H), 1.14 (s, 9H)；MS(ESI)212[M+H]⁺。

The step 2. 7-tert-butyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of 14-3, with N-(2-(thiene-3-yl) ethyl) cyclopropyl carboxamide (16-7) (627mg, Being 3.0mmol) that raw material prepares white solid 697mg, three step gross production rates are 79% (petroleum ether: ethyl acetate=30:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.13 (dd, J=7.3,5.1Hz, 1H), 6.79 (dd, J=9.4,5.1Hz, 1H), 5.14 (s, 1H), 4.24 (dd, J=13.8,5.7Hz, 1H), 3.32 3.09 (m, 1H), 2.82–2.48(m,2H),1.46(s,9H),1.08(s,6H),1.07(s,3H)；Conformer 2:¹H NMR(300 MHz, Chloroform-d) and δ 7.13 (dd, J=7.3,5.1Hz, 1H), 6.79 (dd, J=9.4,5.1Hz, 1H), 4.96 (s, 1 H), 4.47 (dd, J=13.6,6.1Hz, 1H), 3.32 3.09 (m, 1H), 2.82 2.48 (m, 2H), 1.46 (s, 9H), 1.08 (s,6H),1.07(s,3H)；MS(ESI)318[M+Na]⁺。

The step 3. 2-bromo-7-tert-butyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with the 7-tert-butyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 tertiary fourth of (5H)-carboxylic acid Ester (14-9) (697mg, 2.4mmol) is that raw material prepares 694mg colorless oil through bromo, and productivity is 77% (stone Oil ether: ethyl acetate=30:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ6.77(s,1H),5.02(s,1H),4.42(dd, J=13.7,6.4Hz, 1H), 3.33 3.05 (m, 1H), 2.76 2.39 (m, 2H), 1.46 (s, 9H), 1.06 (s, 9H)；Structure As isomer 2:¹H NMR (300MHz, Chloroform-d) δ 6.74 (s, 1H), 4.83 (s, 1H), 4.20 (dd, J= 13.9,6.4Hz,1H),3.33–3.05(m,1H),2.76–2.39(m,2H),1.46(s,9H),1.06(s,9H)；MS (ESI)396[M+Na]⁺。

The step 4. 7-tert-butyl group-2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with the 2-bromo-7-tert-butyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-9) (694mg, 1.9mmol) is that raw material prepares pale yellow oil 223mg, and productivity is 37% (petroleum ether: ethyl acetate=8:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ9.85(s,1H),7.49(s,1H),5.20(s,1 H), 4.50 (dd, J=13.7,6.2Hz, 1H), 3.37 3.08 (m, 1H), 2.86 2.57 (m, 2H), 1.47 (s, 9H), 1.09 (s,9H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.47(s,1H),5.00 (s, 1H), 4.29 (dd, J=14.1,5.6Hz, 1H), 3.37 3.08 (m, 1H), 2.86 2.57 (m, 2H), 1.47 (s, 9H), 1.09(s,9H)；MS(ESI)346[M+Na]⁺。

-4,7-dihydro-thiophene is also for the step 5. 7-tert-butyl group-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base) [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with the 7-tert-butyl group-2-formoxyl-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-9) (223mg, 0.69mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (127mg, 0.69mmol) is that raw material prepares light brown foam shape solid 313mg, productivity be 93% (petroleum ether: Ethyl acetate=3:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 10.42 (s, 1H), 7.64 (d, J=8.9Hz, 1 H), 7.60 (d, J=9.6Hz, 1H), 7.40 (s, 1H), 5.21 (s, 1H), 4.52 (dd, J=12.2,7.3Hz, 1H), 4.02 (s,3H),3.38–3.09(m,1H),2.88–2.53(m,2H),1.48(s,9H),1.13(s,9H)；Conformer 2:¹H NMR (300MHz, Chloroform-d) δ 10.42 (s, 1H), 7.64 (d, J=8.9Hz, 1H), 7.60 (d, J=9.6Hz, 1H), 7.40 (s, 1H), 5.01 (s, 1H), 4.30 (dd, J=13.3,4.7Hz, 1H), 4.02 (s, 3H), 3.38 3.09 (m, 1 H),2.88–2.53(m,2H),1.48(s,9H),1.13(s,9H)；MS(ESI)488[M+H]⁺。

Step 6. 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Acid methyl ester:

According to preparing the operational approach of compound 4-1, with the 7-tert-butyl group-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] miaow Azoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-13) (313mg, 0.64mmol) is Raw material deprotection base obtains yellowish-brown powder 234mg, and productivity is 94% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.40 (s, 1H), 7.63 (dd, J=15.5,1.8Hz, 1H), 7.60 (dd, J=15.8,1.7Hz, 1H), 7.39 (s, 1H), 4.02 (s, 3H), 3.84 (s, 1H), 3.43 3.31 (m, 1H), 2.98–2.85(m,1H),2.80–2.58(m,2H),1.13(s,9H)；MS(ESI)388[M+H]⁺。

Step 7. 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Amide:

According to preparing the operational approach of compound I-1, with 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base) to be that raw material ammonia solution obtains yellowish-brown for-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (4-13) (234mg, 0.60mmol) Color powder 144mg, productivity is 65% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆)δ9.08(brs,1H),7.88(brs,1H),7.67(brs,1H),7.56(dd, J=8.4,2.5Hz, 1H), 7.52 (dd, J=8.0,2.5Hz, 1H), 3.73 (s, 1H), 3.21 (t, J=4.0Hz, 1H), 2.84–2.70(m,1H),2.66–2.54(m,2H),1.07(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δ (165.40,158.71 d, J=235.0Hz, C-F), 149.29,140.61,139.06,138.61,136.02,128.94, 123.26,110.34 (d, J=30.3Hz, CH-CF), 101.80 (d, J=25.4Hz, CH-CF), 64.77,43.02, 35.89,27.42,27.35；HRMS(ESI):m/z Calcd.For C₁₉H₂₂FN₄OS[M+H]⁺:373.1498；Found: 373.1490。

Embodiment 24. (S)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] miaow Azoles-4-Methanamide, the preparation of I-24:

Step 1. (S)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methyl formate:

By 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (4-13) chiral preparation liquid phase separation obtains white powdery solids, and its absolute configuration is determined by single crystal X-ray diffraction. Separation condition is: chromatographic column model: Chiralpak As；Chromatographic column specification: 0.46cm I.D. × 25cm L；Sample size: 5 μL；Flowing phase: CO₂/ IPA/DEA=60/40/0.1 (v/v/v)；Flow velocity: 2.5ml/min；Testing conditions: UV λ=254 nm；Column temperature: 35 DEG C.

¹H NMR (300MHz, Chloroform-d) δ 10.40 (s, 1H), 7.63 (dd, J=15.5,1.8Hz, 1H), 7.60 (dd, J=15.8,1.7Hz, 1H), 7.39 (s, 1H), 4.02 (s, 3H), 3.84 (s, 1H), 3.43 3.31 (m, 1H), 2.98–2.85(m,1H),2.80–2.58(m,2H),1.13(s,9H)；MS(ESI)388[M+H]⁺；–169(c, 0.2,MeOH)；t_R=3.60min.

Step 2. (S)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methanamide:

According to preparing the operational approach of compound I-1, with (S)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base) to be that raw material ammonia solution obtains white for-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (S-4-13) (200mg, 0.52mmol) Color powder 125mg, productivity is 65% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆)δ9.08(brs,1H),7.88(brs,1H),7.67(brs,1H),7.56(dd, J=8.4,2.5Hz, 1H), 7.52 (dd, J=8.0,2.5Hz, 1H), 3.73 (s, 1H), 3.21 (t, J=4.0Hz, 1H), 2.84–2.70(m,1H),2.66–2.54(m,2H),1.07(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δ (165.40,158.71 d, J=235.0Hz, C-F), 149.29,140.61,139.06,138.61,136.02,128.94, 123.26,110.34 (d, J=30.3Hz, CH-CF), 101.80 (d, J=25.4Hz, CH-CF), 64.77,43.02, 35.89,27.42,27.35；HRMS(ESI):m/z Calcd.For C₁₉H₂₂FN₄OS[M+H]⁺:373.1498；Found: 373.1490；–191(c,0.08,MeOH)；Ee > 99%；t_R=12.69min, chromatographic condition is: chromatographic column model: Chiralpak AD-H；Chromatographic column specification: 0.40cm I.D. × 25cm L；Sample size: 5 μ L；Flowing phase: Hexane/IPA/DEA=70/30/0.05 (v/v/v)；Flow velocity: 0.40ml/min；Testing conditions: UV λ=254nm；Post Temperature: 20 DEG C.

Embodiment 25. (R)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] miaow Azoles-4-Methanamide, the preparation of I-25:

Step 1. (R)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methyl formate:

Separation condition such as S-4-13.

¹H NMR (300MHz, Chloroform-d) δ 10.40 (s, 1H), 7.63 (dd, J=15.5,1.8Hz, 1H), 7.60 (dd, J=15.8,1.7Hz, 1H), 7.39 (s, 1H), 4.02 (s, 3H), 3.84 (s, 1H), 3.43 3.31 (m, 1H), 2.98–2.85(m,1H),2.80–2.58(m,2H),1.13(s,9H)；MS(ESI)388[M+H]⁺；141(c,0.2, MeOH)；t_R=1.93min.

Step 2. (R)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1H-benzo [d] imidazoles-4- Methanamide:

According to preparing the operational approach of compound I-1, with (R)-2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base) to be that raw material ammonia solution obtains white for-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (R-4-13) (200mg, 0.52mmol) Color powder 125mg, productivity is 65% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆)δ9.08(brs,1H),7.88(brs,1H),7.67(brs,1H),7.56(dd, J=8.4,2.5Hz, 1H), 7.52 (dd, J=8.0,2.5Hz, 1H), 3.73 (s, 1H), 3.21 (t, J=4.0Hz, 1H), 2.84–2.70(m,1H),2.66–2.54(m,2H),1.07(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δ (165.40,158.71 d, J=235.0Hz, C-F), 149.29,140.61,139.06,138.61,136.02,128.94, 123.26,110.34 (d, J=30.3Hz, CH-CF), 101.80 (d, J=25.4Hz, CH-CF), 64.77,43.02, 35.89,27.42,27.35；HRMS(ESI):m/z Calcd.For C₁₉H₂₂FN₄OS[M+H]⁺:373.1498；Found: 373.1490；190(c,0.09,MeOH)；Ee=98%；t_R=13.42min, chromatographic condition is: chromatographic column model: Chiralpak AD-H；Chromatographic column specification: 0.40cm I.D. × 25cm L；Sample size: 5 μ L；Flowing phase: Hexane/IPA/DEA=70/30/0.05 (v/v/v)；Flow velocity: 0.40ml/min；Testing conditions: UV λ=254nm；Post Temperature: 20 DEG C.

The fluoro-2-of embodiment 26. 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles -4-Methanamide, the preparation of I-26:

Step 1. 3-methyl-N-(2-(thiene-3-yl) ethyl) butyramide:

According to preparing the operational approach of 16-1, with 2-(thiene-3-yl) ethylamine hydrochloride (492mg, 3.0mmol), 3-Methylbutanoyl chloride (434mg, 3.6mmol) is that raw material prepares light yellow oil 627mg, and productivity is 99%, directly Tap into and react into next step.

¹H NMR (300MHz, Chloroform-d) δ 7.28 (dd, J=4.9,3.0Hz, 1H), 7.01 6.98 (m, 1H), 6.95 (dd, J=1.3Hz, 1H), 5.44 (brs, 1H), 3.57 3.48 (m, 2H), 2.84 (t, J=6.8Hz, 2H), 2.19 2.01 (m, 1H), 2.01 1.96 (m, 2H), 0.92 (d, J=6.4Hz, 6H)；MS(ESI)212[M+H]⁺。

Step 2. 7-isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of 14-3, with N-(2-(thiene-3-yl) ethyl) isobutyramide (16-8) (627mg, 3.0 Mmol) being that raw material prepares white solid 816mg, three step gross production rates are 92% (petroleum ether: ethyl acetate=30:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.1Hz, 1H), 6.76 (d, J= 5.1Hz,1H),5.48–5.33(m,1H),4.23–4.04(m,1H),3.13–2.95(m,2H),1.89–1.66(m,3H), (1.47 s, 9H), 1.47 (d, J=1.6Hz, 10H), 1.04 (d, J=6.2Hz, 3H), 0.96 (d, J=6.3Hz, 3H)；Structure As isomer 2:¹H NMR (300MHz, Chloroform-d) δ 7.13 (d, J=5.1Hz, 1H), 6.76 (d, J=5.1Hz, 1H),5.29–5.15(m,1H),4.43–4.27(m,1H),2.84–2.64(m,2H),1.89–1.66(m,3H),1.47(s, 9H), 1.04 (d, J=6.2Hz, 3H), 1.04 (d, J=6.2Hz, 3H), 0.96 (d, J=6.3Hz, 3H)；MS(ESI) 318[M+Na]⁺。

Step 3. 2-bromo-7-isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 7-isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 tertiary fourth of (5H)-carboxylic acid Ester (14-10) (816mg, 2.8mmol) is that raw material prepares 664mg colorless oil through bromo, and productivity is 64% (stone Oil ether: ethyl acetate=30:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.38–5.20(m,1H), 4.20 3.92 (m, 1H), 3.19 2.89 (m, 2H), 1.73 (hept, J=6.3Hz, 1H), 1.47 (s, 9H), 1.02 (d, J= 6.3Hz, 3H), 0.95 (d, J=6.4Hz, 3H)；Conformer 2:¹H NMR(300MHz,Chloroform-d)δ (6.71 s, 1H), 5.16 4.97 (m, 1H), 4.42 4.23 (m, 1H), 2.80 2.42 (m, 2H), 1.73 (hept, J=6.3Hz, 1H), 1.47 (s, 9H), 1.02 (d, J=6.3Hz, 3H), 0.95 (d, J=6.4Hz, 3H)；MS(ESI)396[M+Na]⁺。

Step 4. 2-formoxyl-7-isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with 2-bromo-7-isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-10) (664mg, 1.8mmol) is that raw material prepares yellow oil 400mg, and productivity is 69% (petroleum ether: ethyl acetate=8:1).

Conformer 1:¹H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.45(brs, 1H),4.24–4.12(m,1H),3.19–2.70(m,2H),2.68–2.52(m,1H),1.86–1.70(m,2H),1.48(s, 9H), 1.05 (d, J=6.4Hz, 3H), 0.97 (d, J=6.4Hz, 3H)；Conformer 2:¹H NMR(300MHz, Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.25(brs,1H),4.44–4.33(m,1H),3.19–2.70(m, 2H), 2.68 2.52 (m, 1H), 1.86 1.70 (m, 2H), 1.48 (s, 9H), 1.05 (d, J=6.4Hz, 3H), 0.97 (d, J=6.4Hz, 3H)；MS(ESI)346[M+Na]⁺。

-7-isobutyl group-4,7-dihydro-thiophene is also for step 5. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base) [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-7-isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-10) (400mg, 1.24mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (228mg, 1.24mmol) is that raw material prepares faint yellow solid 553mg, and productivity is 91% (petroleum ether: acetic acid Ethyl ester=3:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.64 (dd, J=9.2,2.4Hz, 1H), 7.62 (dd, J=9.2,2.4Hz, 1H), 7.39 (s, 1H), 5.45 (brs, 1H), 4.30 4.12 (m, 1H), 4.02 (s, 3H), 2.91–2.70(m,2H),2.69–2.54(m,1H),1.95–1.68(m,2H),1.67–1.50(m,1H),1.50(s,9H), 1.06 (d, J=6.4Hz, 3H), 1.01 (d, J=6.5Hz, 3H)；Conformer 2:¹H NMR(300MHz, Chloroform-d) δ 7.64 (dd, J=9.2,2.4Hz, 1H), 7.62 (dd, J=9.2,2.4Hz, 1H), 7.39 (s, 1H), 5.25(brs,1H),4.50–4.32(m,1H),4.02(s,3H),3.20–2.96(m,2H),2.69–2.54(m,1H), 1.95 1.68 (m, 2H), 1.67 1.50 (m, 1H), 1.50 (s, 9H), 1.06 (d, J=6.4Hz, 3H), 1.01 (d, J=6.5 Hz,3H)；MS(ESI)488[M+H]⁺。

The fluoro-2-of step 6. 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Acid methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7- Isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-14) (553mg, 1.13mmol) is former Material deprotection base obtains white powder 374mg, and productivity is 85% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.62 (dd, J=13.3,2.4Hz, 1H), (7.59 dd, J=14.1,2.4Hz, 1H), 7.38 (s, 1H), 4.17 4.03 (m, 1H), 4.02 (s, 3H), 3.34 (ddd, J= 13.0,5.3,3.7Hz, 1H), 3.01 (ddd, J=13.0,8.7,5.6Hz, 1H), 2.75 2.65 (m, 2H), 2.11 1.91 (m, 1H), 1.67 1.62 (m, 2H), 1.02 (d, J=4.2Hz, 3H), 1.00 (d, J=4.3Hz, 3H)；MS(ESI)388 [M+H]⁺。

The fluoro-2-of step 7. 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine -2-base) to be that raw material ammonia solution obtains faint yellow for-1H-benzo [d] imidazoles-4-methyl formate (4-14) (374mg, 0.97mmol) Powder 241mg, productivity is 67% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆)δ9.01(brs,1H),7.92(brs,1H),7.62(s,1H),7.53(dd,J =10.7,2.6Hz, 1H), 7.50 (dd, J=8.5,2.6Hz, 1H), 3.94 (dd, J=10.1,4.3Hz, 1H), 3.11 (dt, J =13.1,5.4Hz, 1H), 2.81 (ddd, J=13.4,8.7,5.4Hz, 1H), 2.63-2.52 (m, 2H), 1.99 1.89 (m, 1 H), 1.58 (ddd, J=14.2,9.7,4.7Hz, 1H), 1.48 (ddd, J=13.6,9.2,4.3Hz, 1H), 0.93 (d, J=6.6 Hz, 3H), 0.92 (d, J=6.6Hz, 3H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.46,158.6 (d, J= 237.5Hz, C-F), 149.45,145.03,136.18,129.01,128.49,122.76,110.11 (d, J=25.8Hz, CH-CF), 102.11 (d, J=23.4Hz, CH-CF), 52.54,46.85,41.76,26.57,24.56,23.87,21.91； HRMS(ESI):m/z Calcd.For C₁₉H₂₂FN₄OS[M+H]⁺:373.1498；Found:373.1490.

The fluoro-2-of embodiment 27. 6-(5-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-27:

Step 1. 3-(2-nitropropyl-1-alkene-1-base) thiophene:

Under ice bath, to thiophene-3-formaldehyde (18) (2.24g, 20.0mmol) and the mixing of nitroethane (60mL) Thing adds ammonium acetate (2.00g, 26.0mmol), refluxes two hours, by reactant liquor concentrating under reduced pressure, be redissolved in 50mL In ethyl acetate, washing successively, saturated common salt is washed, and anhydrous sodium sulfate is dried, and filters concentrating under reduced pressure and prepares Light brown solid 2.87g, productivity is 85%.

¹H NMR (300MHz, Chloroform-d) δ 8.09 (s, 1H), 7.60 (brs, 1H), 7.44 (dd, J=5.0,3.0 Hz, 1H), 7.27 (d, J=4.6Hz, 1H), 2.50 (s, 3H)；MS(ESI)170[M+H]⁺。

Step 2. (1-(thiene-3-yl) propyl group-2-base) t-butyl carbamate:

At 0 DEG C, anhydrous toward dissolved with in 3-(2-nitropropyl-1-alkene-1-base) thiophene (19) (1.70g, 10.0mmol) Oxolane (10mL) solution adds lithium aluminium hydride reduction tetrahydrofuran solution (2.5M) (12.5mL), is slowly increased to 60 DEG C are reacted 1 hour, and ice bath cools down, and are slowly added to five aqueous sodium persulfate cancellation, and dichloromethane extracts (50mL × 3), Washing, saturated common salt is washed, and retains organic layer.At 0 DEG C toward organic layer is slowly added to Bis(tert-butoxycarbonyl)oxide (1.78g, 10.0mmol), moving to be stirred at room temperature 1 hour, decompression boils off major part solvent, obtains pale yellow oil 1.33 through column chromatography G, productivity is 55%.

¹H NMR (300MHz, Chloroform-d) δ 7.26 (dd, J=4.7,2.9Hz, 1H), 7.00 6.96 (m, 1H), (6.94 dd, J=4.9,1.1Hz, 1H), 4.36 (brs, 1H), 3.91 (brs, 1H), 2.78 (qd, J=14.1,6.1Hz, 1H), (1.43 s, 9H), 1.09 (d, J=6.6Hz, 3H)；MS(ESI)264[M+Na]⁺。

Step 3. 5-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

To dissolved with the tert-butyl group (1-(thiene-3-yl) propyl group-2-base) carbamate (21) (500mg, 2.07mmol) Adding paraformaldehyde (150mg, 5.00mmol) in toluene solution, one is hydrated p-methyl benzenesulfonic acid (19mg, 0.10mmol), Overnight, decompression boils off major part solvent, and thick product obtains pale yellow oil 257mg through column chromatography, and productivity is 49% in backflow.

¹H NMR (300MHz, Chloroform-d) δ 7.15 (d, J=5.0Hz, 1H), 6.77 (d, J=5.0Hz, 1H), 4.97 (d, J=16.9Hz, 1H), 4.75 (brs, 1H), 4.19 (d, J=16.8Hz, 1H), 2.93 (dd, J=16.0,6.5Hz, 1H), 2.50 (d, J=15.6Hz, 1H), 1.48 (s, 9H), 1.12 (d, J=6.9Hz, 3H)；MS(ESI)276 [M+H]⁺。

Step 4. 2-bromo-5-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 5-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (14-11) (257mg, 1.02mmol) is that raw material prepares 249mg pale yellow oil through bromo, and productivity is 75% (petroleum ether: ethyl acetate=30:1).

¹H NMR (300MHz, Chloroform-d) δ 6.73 (s, 1H), 4.84 (d, J=17.2Hz, 1H), 4.70 (brs, 1 H), 4.07 (d, J=17.0Hz, 1H), 2.87 (ddt, J=16.1,5.5,2.4Hz, 1H), 2.40 (dt, J=15.9,2.4Hz, 1 H), 1.47 (s, 9H), 1.12 (d, J=6.9Hz, 3H)；MS(ESI)354[M+Na]⁺。

Step 5. 2-formoxyl-5-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-3, with 2-bromo-5-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)- Carboxylic acid tert-butyl ester (12-11) (249mg, 0.98mmol) is that raw material prepares pale yellow oil 134mg, productivity It is 48% (petroleum ether: ethyl acetate=8:1).

¹H NMR (300MHz, Chloroform-d) δ 9.85 (s, 1H), 7.48 (s, 1H), 5.04 (d, J=18.3Hz, 1 H), 4.78 (brs, 1H), 4.24 (d, J=18.3Hz, 1H), 2.97 (dd, J=15.8,5.9Hz, 1H), 2.54 (d, J=15.9 Hz, 1H), 1.49 (s, 9H), 1.12 (d, J=6.9Hz, 3H)；MS(ESI)282[M+H]⁺。

Step 6. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-5-methyl-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-5-methyl-4,7-dihydro-thiophene also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters (2-11) (134mg, 0.47mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (86mg, 0.47mmol) is that raw material prepares faint yellow solid 187mg, and productivity is 89% (petroleum ether: acetic acid Ethyl ester=3:1).

¹H NMR (300MHz, Chloroform-d) δ 10.40 (s, 1H), 7.63 (dd, J=13.4,2.1Hz, 1H), 7.60 (dd, J=14.4,2.3Hz, 1H), 7.38 (s, 1H), 5.11-4.97 (d, J=16.0,1H), 4.80 (brs, 1H), 4.31-4.19 (m, 1H), 4.02 (s, 3H), 2.97 (d, J=15.8,5.7Hz, 1H), 2.55 (d, J=15.6Hz, 1H), 1.50 (s, 9H), 1.16 (d, J=7.0Hz, 3H)；MS(ESI)446[M+H]⁺。

The fluoro-2-of step 7. 6-(5-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-5- Methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-15) (187mg, 0.42mmol) is raw material Deprotection base obtains white powder 119mg, and productivity is 82% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.38 (brs, 1H), 7.63 (dd, J=12.5,2.3Hz, 1H), (7.59 dd, J=14.0,2.3Hz, 1H), 7.37 (s, 1H), 4.16 (s, 2H), 4.02 (s, 3H), 3.13-3.00 (m, 1H), (2.79 dd, J=16.2,4.0Hz, 1H), 2.38 (dd, J=16.1,10.1Hz, 1H), 1.30 (d, J=6.4Hz, 3H)；MS (ESI)346[M+H]⁺。

The fluoro-2-of step 8. 6-(5-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(5-methyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (4-15) (119mg, 0.34mmol) is that raw material ammonia solution obtains faint yellow powder End 82mg, productivity is 73% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆)δ8.96(brs,1H),7.89(brs,1H),7.67(s,1H),7.57–7.45 (m, 2H), 4.06 (d, J=28.9,1H), 4.01 (d, J=29.1,1H), 2.99 (brs, 1H), 2.74 (dd, J=16.3,3.9 Hz, 1H), 2.41 2.25 (m, 1H), 1.19 (d, J=6.3Hz, 3H)；HRMS(ESI):m/z Calcd.For C₁₆H₁₆FN₄OS[M+H]⁺:331.1029；Found:311.1023.

The fluoro-2-of embodiment 28. 6-(3-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-28:

Step 1. 5-tert-butyl group 3-ethyl 2-amino-6,7-dihydro-thiophene also [3,2-c] pyridine-3,5 (4H)-dicarboxylic ester:

To dissolved with N-tertbutyloxycarbonyl-4-piperidones (22) (9.95g, 50.0mmol) and cyan-acetic ester (5.65g, 50.0mmol) dehydrated alcohol (250mL) solution in add 1.60g sulfur powder and 4.35g morpholine, reflux 1 hour, Concentrating under reduced pressure, thick product is in ethanol through being recrystallized to give light tan solid 13.7g, and productivity is 84%.

¹H NMR (300MHz, Chloroform-d) δ 6.01 (brs, 2H), 4.35 (brs, 2H), 4.26 (q, J=7.1Hz, 2H), 3.61 (t, J=5.9Hz, 2H), 2.80 (t, J=5.9Hz, 2H), 1.47 (s, 9H), 1.33 (t, J=7.1Hz, 3H)； MS(ESI)327[M+H]⁺。

Step 2. 5-tert-butyl group 3-ethyl 6,7-dihydro-thiophene also [3,2-c] pyridine-3,5 (4H)-dicarboxylic ester:

At 0 DEG C, toward dissolved with 5-tert-butyl group 3-ethyl 2-amino-6,7-dihydro-thiophene also [3,2-c] pyridine-3,5 (4H)-dicarboxylic acids Oxolane (30mL) solution of ester (23) (6.10g, 18.7mmol) is slowly added dropwise amyl nitrite (5.54 ML, 41.2mmol), move to after dropping be stirred at room temperature 30 minutes, be continuously heating to backflow, react two hours, Being down to room temperature, concentrating under reduced pressure, thick product obtains white solid 1.88 through column chromatography (petroleum ether: ethyl acetate=10:1) G, productivity is 32%.

¹H NMR (300MHz, Chloroform-d) δ 7.97 (s, 1H), 4.60 (s, 2H), 4.29 (q, J=7.1Hz, 2H), (3.66 t, J=5.8Hz, 2H), 2.98 (t, J=5.9Hz, 2H), 1.48 (s, 9H), 1.35 (m, J=7.1Hz, 3H)；MS (ESI)334[M+Na]⁺。

Step 3. 3-(methylol)-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

It is slowly added dropwise uncle 5-in anhydrous tetrahydro furan (5mL) suspension of lithium borohydride (530mg, 24.0mmol) Butyl 3-ethyl 6,7-dihydro-thiophene also [3,2-c] pyridine-3,5 (4H)-dicarboxylic esters (24) (1.88g, 6.0mmol) Toluene (10mL) solution, is heated to 100 DEG C and refluxes 5 hours.Concentrating under reduced pressure, thick product through column chromatography (petroleum ether: Ethyl acetate=4:1) obtain colorless oil 1.02g, productivity is 63%.

¹H NMR (300MHz, Chloroform-d) δ 7.08 (s, 1H), 4.61 (s, 2H), 4.58 (d, J=4.6Hz, 2H), (3.69 t, J=5.8Hz, 2H), 2.68 (t, J=5.8Hz, 2H), 1.48 (s, 9H)；MS(ESI)292[M+Na]⁺。

Step 4. 3-(chloromethyl)-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

At 0 DEG C, past dissolved with 3-(methylol)-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (25) (1.02 G, 3.8mmol) and dichloromethane (10mL) solution of triethylamine (1.06mL, 7.6mmol) in be slowly added dropwise first Sulfonic acid chloride (866mg, 7.6mmol), is stirred at room temperature 5 hours.Concentrating under reduced pressure, thick product through column chromatography (petroleum ether: Ethyl acetate=8:1) obtain white solid 817mg, productivity is 75%.

¹H NMR (300MHz, Chloroform-d) δ 7.15 (s, 1H), 4.60 (s, 2H), 4.50 (s, 2H), 3.71 (t, J= 5.8Hz, 2H), 2.71 (t, J=5.7Hz, 2H), 1.48 (s, 9H)；MS(ESI)310[M+Na]⁺。

Step 5. 3-methyl-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

Toward 3-(chloromethyl)-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (26) (817mg, 2.8mmol) Methanol (10mL) solution in add Raney's nickel (80mg), nitrogen protect, replacing hydrogen three times, in atmosphere of hydrogen In be heated to 60 DEG C, reflux 4 hours.Be cooled to room temperature, filter, concentrating under reduced pressure, thick product through column chromatography (petroleum ether: Ethyl acetate=20:1) obtain colorless oil 636mg, productivity is 90%.

¹H NMR (300MHz, Chloroform-d) δ 6.76 (s, 1H), 4.60 (s, 2H), 3.69 (t, J=5.7Hz, 2H), 2.56 (t, J=5.7Hz, 2H), 2.11 (s, 3H), 1.48 (s, 9H)；MS(ESI)276[M+Na]⁺。

Step 6. 2-bromo-3-methyl-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 12-1, with 3-methyl-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (14-12) (636mg, 2.5mmol) is that raw material prepares 586mg faint yellow solid through bromo, and productivity is 70% (stone Oil ether: ethyl acetate=30:1).

¹H NMR (300MHz, Chloroform-d) δ 4.49 (s, 2H), 3.67 (t, J=5.6Hz, 2H), 2.53 (t, J= 5.5Hz,2H),2.04(s,3H),1.48(s,9H)；MS(ESI)354[M+Na]⁺。

Step 7. 2-formoxyl-3-methyl-67-dihydro-thiophene also [32-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-1, with 2-bromo-3-methyl-6,7-dihydro-thiophene also [3,2-c] pyridine-5 (4H)- Carboxylic acid tert-butyl ester (12-12) (586mg, 1.77mmol) is that raw material prepares yellow oil 379mg, and productivity is 7% (petroleum ether: ethyl acetate=8:1).

¹H NMR (300MHz, Chloroform-d) δ 10.02 (s, 1H), 4.64 (s, 2H), 3.70 (t, J=6.0Hz, 2H), 2.60 (t, J=5.9Hz, 2H), 2.42 (s, 3H), 1.48 (s, 9H)；MS(ESI)304[M+Na]⁺。

Step 8. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-3-methyl-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-3-methyl-6,7-dihydro-thiophene also [3,2-c] pyridine -5 (4H)-carboxylic acid tert-butyl esters (2-12) (379mg, 1.35mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (248mg, 1.35mmol) is that raw material prepares white solid 445mg, and productivity is 74% (petroleum ether: acetic acid second Ester=3:1).

¹H NMR (300MHz, Chloroform-d) δ 10.39 (brs, 1H), 7.66 (dd, J=8.9,2.4Hz, 1H), 7.58 (dd, J=9.6,2.4Hz, 1H), 4.66 (s, 2H), 4.02 (s, 3H), 3.74 (t, J=5.6Hz, 2H), 2.64 (t, J=5.6 Hz,2H),2.52(s,3H),1.50(s,9H)；MS(ESI)446[M+H]⁺。

The fluoro-2-of step 9. 6-(3-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-3- Methyl-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-16) (445mg, 1.00mmol) is raw material Deprotection base obtains white powder 226mg, and productivity is 65% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.39 (brs, 1H), 7.65 (dd, J=8.9,2.3Hz, 1H), 7.57 (dd, J=9.7,2.4Hz, 1H), 4.07 (s, 2H), 4.01 (s, 3H), 3.18 (t, J=5.8Hz, 2H), 2.59 (t, J=5.8 Hz,2H),2.51(s,3H)；MS(ESI)346[M+H]⁺。

The fluoro-2-of step 10. 6-(3-methyl-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(3-methyl-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (4-16) (226mg, 0.65mmol) is that raw material ammonia solution obtains faint yellow powder End 151mg, productivity is 70% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆) δ 9.14 (brs, 1H), 7.92 (brs, 1H), 7.55 (dd, J=10.7,2.6 Hz, 1H), 7.47 (dd, J=8.5,2.6Hz, 1H), 3.92 (s, 2H), 3.17 (s, 2H), 3.00 (t, J=5.7Hz, 2H), 2.46(s,3H)；¹³C NMR(100MHz,DMSO-d₆) δ 165.65,158.43 (d, J=234.4Hz, C-F), 149.83, (138.36,138.23,137.78,136.51,122.68,110.12 d, J=26.1Hz, CH-CF), 102.30 (d, J=27.0 Hz,CH-CF),44.56,42.63,25.27,13.76；HRMS(ESI):m/z Calcd.For C₁₆H₁₆FN₄OS[M+H]⁺: 331.1029；Found:331.1023.

The fluoro-2-of embodiment 29. 6-(4,5,6,7-tetrahydro-thiazoles also [5,4-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide, The preparation of I-29:

Step 1. 2-amino-6,7-thiazoline [4,5-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

To hexamethylene (25mL) solution dissolved with N-tertbutyloxycarbonyl-4-piperidones (22) (9.95g, 50.0mmol) Middle addition pyrrolidine (4.35mL, 52.0mmol) and hydration p-methyl benzenesulfonic acid (48mg, 2.5mmol), connect point Hydrophone refluxes 50 minutes, is cooled to room temperature, evaporated under reduced pressure hexamethylene, is redissolved in 15mL absolute methanol, then to mixing Adding 1.60g sulfur powder in thing, be stirred at room temperature 20 minutes, water-bath cools down, and slowly adds the 50% of 4.2mL in reactant liquor Cyanamide aqueous solution, continues stirring 3 hours under water-bath, remove water-bath and continue stirring 17 hours, separate out white solid, add Water 40mL, filters, washes to obtain 8.00g white solid, and productivity is 63%.

¹H NMR(300MHz,DMSO-d₆) δ 6.78 (s, 2H), 4.28 (d, J=2.4Hz, 2H), 3.55 (t, J=5.6 Hz, 2H), 2.41 (d, J=6.3Hz, 2H), 1.40 (s, 9H)；MS(ESI)256[M+H]⁺。

Step 2. 2-bromo-6,7-thiazoline also [4,5-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

At 0 DEG C, toward 2-amino-6,7-thiazoline [4,5-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (27) (1.62g, 5.0mmol) DMF (15mL) solution in, add copper bromide (1.70g, 7.6mmol), stir 5 minutes, At such a temperature, then in reactant liquor, it is slowly added dropwise 1.5mL amyl nitrite, after dropping, is slowly increased to room temperature Continue stirring 4 hours.Reactant liquor dichloromethane extracts (50mL × 3), and organic layer washes with water successively, saturated aqueous common salt Washing, retain organic layer, be dried with anhydrous sodium sulfate, filter concentrating under reduced pressure, thick product is through column chromatography (petroleum ether: acetic acid second Ester=20:1) obtain weak yellow liquid 648mg, productivity is 40%.

¹H NMR (300MHz, Chloroform-d) δ 4.56 (brs, 2H), 3.72 (t, J=5.6Hz, 2H), 2.85 (t, J= 5.6Hz,2H),1.48(s,9H)；MS(ESI)319[M+H]⁺。

Step 3. 2-formoxyl-6,7-thiazoline also [4,5-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester:

-78 DEG C, under nitrogen protection, toward 2-bromo-6,7-thiazoline also [4,5-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (12-13) Anhydrous tetrahydro furan (30mL) solution of (648mg, 2.0mmol) is slowly added dropwise the tetrahydrochysene of 1.0mL n-BuLi Tetrahydrofuran solution (2.5M), continues stirring 5 minutes.The anhydrous DMF of 176mg is added in reactant liquor, After stirring 1 hour, add 1mL shrend and go out.Reactant liquor is extracted with ethyl acetate (50mL × 3), and organic layer is used successively Water, saturated aqueous common salt wash, and retain organic layer, are dried with anhydrous sodium sulfate, filter concentrating under reduced pressure, and thick product is through column chromatography (petroleum ether: ethyl acetate=10:1) obtains water white transparency oily thing 415mg, and productivity is 77%.

¹H NMR (300MHz, Chloroform-d) δ 9.92 (s, 1H), 4.75 (s, 2H), 3.79 (t, J=5.8Hz, 2H), (2.99 t, J=5.9Hz, 2H), 1.49 (s, 9H)；MS(ESI)269[M+H]⁺。

Step 4. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-6,7-thiazoline also [5,4-c] pyridine -5 (4H)-carboxylic acid tert-butyl esters:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-6,7-thiazoline also [4,5-c] pyridine-5 (4H)-carboxylic acid The tert-butyl ester (2-13) (415mg, 1.55mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (285mg, 1.55mmol) it is that raw material prepares white solid 228mg, productivity is 34% (petroleum ether: ethyl acetate=2:1).

¹H NMR (300MHz, Chloroform-d) δ 11.04 (brs, 1H), 7.71 (dd, J=4.5,2.5Hz, 1H), 7.68 (dd, J=3.9,2.5Hz, 1H), 4.75 (s, 2H), 4.04 (s, 3H), 3.81 (t, J=5.5Hz, 2H), 2.96 (t, J=5.5 Hz,2H),1.51(s,9H)；MS(ESI)433[M+H]⁺。

The fluoro-2-of step 5. 6-(4,5,6,7-tetrahydro-thiazoles also [5,4-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-carboxylate methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-6,7- Thiazoline also [5,4-c] pyridine-5 (4H)-carboxylic acid tert-butyl ester (3-17) (228mg, 0.53mmol) is that raw material deprotects base Obtaining light brown powder 103mg, productivity is 59% (dichloromethane: methanol=20:1).

¹H NMR (300MHz, Chloroform-d) δ 7.71 (dd, J=4.9,2.4Hz, 1H), 7.68 (dd, J=3.9,2.4 Hz, 1H), 4.17 (brs, 2H), 4.04 (s, 3H), 3.26 (t, J=5.8Hz, 2H), 2.93 (t, J=5.8Hz, 2H)；MS (ESI)333[M+H]⁺。

The fluoro-2-of step 6. 6-(4,5,6,7-tetrahydro-thiazoles also [5,4-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(4,5,6,7-tetrahydro-thiazoles also [5,4-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-17) (103mg, 0.31mmol) is that raw material ammonia solution obtains light brown powder 65mg, Productivity is 66% (dichloromethane: methanol=8:1).

¹H NMR(300MHz,DMSO-d₆) δ 8.86 (brs, 1H), 8.04 (brs, 1H), 7.64 (dd, J=10.6,2.6 Hz, 1H), 7.49 (dd, J=8.4,2.6Hz, 1H), 4.39 (brs, 2H), 3.41 (t, J=6.2Hz, 2H), 3.06 (t, J= 5.9Hz,2H)；¹³C NMR (101MHz, TFA-d) δ 167.73,161.60 (d, J=253.6Hz, C-F), 150.70, (147.41,142.78,132.11 d, J=12.8Hz, C-CH-CF), 129.52,126.60,119.47 (d, J=10.8Hz, C-CH-CF), 116.01,105.65 (d, J=28.4Hz, CH-CF), 42.37,28.98,22.73；HRMS(ESI):m/z Calcd.For C₁₄H₁₃FN₅OS[M+H]⁺:318.0825；Found:318.0820.

Embodiment 30. 6-fluoro-1-methyl-2-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-30:

Step 1. 2-(6-fluoro-4-(methoxycarbonyl)-1-methyl isophthalic acid H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

Toward 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic In acetone (5mL) solution of tert-butyl acrylate (3-6) (500mg, 1.16mmol), add K₂CO₃(240mg, 1.74mmol) with iodomethane (198mg, 1.39mmol), being stirred overnight, reactant liquor is extracted with ethyl acetate (20mL × 3), Organic layer washes with water successively, and saturated common salt is washed, and retains organic layer, is dried with anhydrous sodium sulfate, filters concentrating under reduced pressure, slightly Product obtains faint yellow solid 222mg through column chromatography (petroleum ether: ethyl acetate=3:1), and productivity is 43%.

¹H NMR (300MHz, Chloroform-d) δ 7.72 (dd, J=10.2,2.4Hz, 1H), 7.37 (s, 1H), 7.24 (dd, J=9.8,2.4Hz, 1H), 4.69 (s, 2H), 4.04 (s, 3H), 3.96 (s, 3H), 3.71 (t, J=5.8Hz, 2H), (2.77 t, J=6.0Hz, 2H), 1.50 (s, 9H)；MS(ESI)446[M+H]⁺。

Step 2. 6-fluoro-1-methyl-2-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1-methyl isophthalic acid H-benzo [d] imidazoles-2- Base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (6-1) (222mg, 0.50mmol) is that raw material goes Protection group obtains white powder 154mg, and productivity is 89% (dichloromethane: methanol=20:1).

¹H NMR (300MHz, Chloroform-d) δ 7.69 (dd, J=10.2,2.5Hz, 1H), 7.35 (s, 1H), 7.20 (dd, J=8.0,2.5Hz, 1H), 4.09 (brs, 2H), 4.03 (s, 3H), 3.94 (s, 3H), 3.16 (t, J=5.8Hz, 2H), (2.71 t, J=5.8Hz, 2H), 1.24 (s, 9H)；MS(ESI)346[M+H]⁺。

Step 3. 6-fluoro-1-methyl-2-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with 6-fluoro-1-methyl-2-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (7-1) (154mg, 0.45mmol) is that raw material ammonia solution obtains white powder 116mg, productivity is 79% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆) δ 9.05 (brs, 1H), 8.00 (brs, 1H), 7.81 (d, J=8.6Hz, 1H), (7.60 s, 1H), 7.58 (d, J=12.9Hz, 1H), 4.01 (brs, 5H), 3.03 (brs, 2H), 2.68 (brs, 2H)；¹³C NMR(125MHz,DMSO-d₆) δ 165.22,158.77 (d, J=237.0Hz, C-F), 149.48,138.10 (d, J= 12.9Hz, C-CH-CF), 136.97,136.02,130.17,127.91,123.36 (d, J=8.6Hz, C-CH-CF), 110.93 (d, J=26.3Hz, CH-CF), 101.49 (d, J=28.6Hz, CH-CF), 44.28,42.66,32.80,26.01；HRMS (ESI):m/z Calcd.For C₁₆H₁₆FN₄OS[M]⁺:331.1029；Found:331.1024.

The embodiment 31. 1-fluoro-2-of ethyl-6-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-first Amide, the preparation of I-31:

Step 1. 2-(1-ethyl-6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] Pyridine-6 (5H)-carboxylic acid tert-butyl ester:

Toward 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic In DMF (5mL) solution of tert-butyl acrylate (3-6) (500mg, 1.16mmol), add Cs₂CO₃ (567mg, 1.74mmol) and bromoethane (253mg, 2.32mmol), 90 DEG C are reacted overnight, reactant liquor second Acetoacetic ester extraction (30mL × 3), organic layer with water, saturated aqueous common salt washing, retains organic layer, uses anhydrous slufuric acid successively Sodium is dried, and filters concentrating under reduced pressure, and thick product obtains faint yellow solid 245 through column chromatography (petroleum ether: ethyl acetate=3:1) Mg, productivity is 46%.

¹H NMR (300MHz, Chloroform-d) δ 7.71 (dd, J=10.4,2.0Hz, 1H), 7.37 (s, 1H), 7.24 (dd, J=10.1,2.4Hz, 1H), 4.68 (s, 2H), 4.41 (d, J=7.2Hz, 2H), 4.04 (s, 3H), 3.72 (t, J=5.1 Hz, 2H), 2.75 (t, J=5.1Hz, 2H), 1.50 (s, 9H)；MS(ESI)460[M+H]⁺。

The step 2. 1-fluoro-2-of ethyl-6-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formic acid Methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(1-ethyl-6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2- Base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (6-2) (245mg, 0.53mmol) is that raw material goes Protection group obtains brown solid 156mg, and productivity is 82% (dichloromethane: methanol=30:1).

¹H NMR (300MHz, Chloroform-d) δ 7.71 (dd, J=10.2,2.4Hz, 1H), 7.36 (d, J=2.0Hz, 1H), 7.24 (dd, J=9.1,3.1Hz, 1H), 4.40 (q, J=7.3Hz, 2H), 4.12 (s, 2H), 4.04 (s, 3H), 3.20 (t, J=5.8Hz, 2H), 2.75 (t, J=5.6Hz, 2H), 1.49 (t, J=7.3Hz, 3H)；MS(ESI)360[M+H]⁺。

The step 3. 1-fluoro-2-of ethyl-6-(4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-formyl Amine:

According to preparing the operational approach of compound I-1, with the 1-fluoro-2-of ethyl-6-(4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-1H-benzo [d] imidazoles-4-methyl formate (7-2) (156mg, 0.43mmol) is that raw material ammonia solution obtains white powder 120mg, productivity is 81% (dichloromethane: methanol=10:1).

¹H NMR(500MHz,DMSO-d₆) δ 9.04 (brs, 1H), 8.03 (brs, 1H), 7.88 (dd, J=8.9,2.6Hz, 1H), 7.60 (dd, J=9.2,2.4Hz, 1H), 7.59 (s, 1H), 4.54 (q, J=7.2Hz, 2H), 4.10 (s, 2H), 3.11 (t, J=5.7Hz, 2H), 2.76 (t, J=5.7Hz, 2H), 1.40 (t, J=7.1Hz, 3H)；¹³C NMR(125MHz, DMSO-d₆) δ 165.11,158.87 (d, J=237.1Hz, C-F), 148.49,137.47,137.08 (d, J=8.8Hz, C-CH-CF), 137.07,135.83,129.43,128.15,123.60 (d, J=8.4Hz, C-CH-CF), 111.16 (d, J= 26.6Hz, CH-CF), 101.56 (d, J=28.2Hz, CH-CF), 43.70,42.21,29.44,25.22,15.17；HRMS (ESI):m/z Calcd.For C₁₇H₁₈FN₄OS[M+H]⁺:345.1185；Found:345.1177.

The fluoro-2-of embodiment 32. 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1-methyl isophthalic acid H-benzo [d] Imidazoles-4-Methanamide, the preparation of I-32:

Step 1. 2-(6-fluoro-4-(methoxycarbonyl)-1-methyl isophthalic acid H-benzo [d] imidazoles-2-base)-7-isobutyl group-4,7-dihydro-thiophene And [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 6-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7-isobutyl group -4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-14) (386mg, 0.79mmol) is prepared by raw material Obtaining white solid 263mg, productivity is 66% (petroleum ether: ethyl acetate=8:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.71 (dd, J=10.2,2.5Hz, 1H), 7.30 (s, 1H), 7.23 (dd, J=7.9,2.5Hz, 1H), 5.53 5.42 (m, 1H), 4.26 4.12 (m, 1H), 4.04 (s, 3H), 3.96(s,3H),3.19–2.96(m,2H),2.63–2.55(m,1H),1.96–1.73(m,1H),1.63–1.54(m,2H), 1.49 (s, 9H), 1.05 (d, J=6.3Hz, 3H), 0.99 (d, J=6.5Hz, 3H)；Conformer 2:¹H NMR(300 MHz, Chloroform-d) δ 7.71 (dd, J=10.2,2.5Hz, 1H), 7.30 (s, 1H), 7.23 (dd, J=7.9,2.5Hz, 1H),5.34–5.21(m,1H),4.47–4.33(m,1H),4.04(s,3H),3.96(s,3H),2.93–2.69(m,2H), 2.69–2.62(m,1H),2.63–2.55(m,1H),1.96–1.73(m,1H),1.63–1.54(m,2H),1.49(s,9H), 1.05 (d, J=6.3Hz, 3H), 0.99 (d, J=6.5Hz, 3H)；MS(ESI)502[M+H]⁺。

The fluoro-2-of step 2. 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1-methyl isophthalic acid H-benzo [d] miaow Azoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1-methyl isophthalic acid H-benzo [d] imidazoles-2- Base)-7-isobutyl group-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (6-3) (263mg, 0.52mmol) Obtaining white solid 192mg for raw material deprotection base, productivity is 91% (dichloromethane: methanol=50:1).

¹H NMR(400MHz,DMSO-d₆) δ 7.72 (dd, J=8.8,2.6Hz, 1H), 7.36 (s, 1H), 7.21 (d, J= 10.4,2.5Hz,1H),4.01(s,3H),3.20(brs,1H),2.83(brs,1H),2.61(brs,2H),1.95(s,1H), 1.66 1.55 (m, 1H), 1.54-1.48 (m, 1H), 0.99 (d, J=6.1Hz, 3H), 0.97 (d, J=6.1Hz, 3H)；MS (ESI)402[M+H]⁺。

The fluoro-2-of step 3. 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-1-methyl isophthalic acid H-benzo [d] miaow Azoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(7-isobutyl group-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine -2-base)-1-methyl isophthalic acid H-benzo [d] imidazoles-4-methyl formate (7-3) (192mg, 0.48mmol) is that raw material ammonia solution obtains White powder 128mg, productivity is 69% (dichloromethane: methanol=10:1).

¹H NMR(400MHz,DMSO-d₆) δ 9.04 (brs, 1H), 7.96 (brs, 1H), 7.79 (dd, J=8.8,2.6Hz, 1H), 7.56 (s, 1H), 7.55 (d, J=10.4,2.5Hz, 1H), 4.00 (s, 3H), 3.13 (brs, 1H), 2.81 (brs, 1H), 2.59 (brs, 2H), 1.94 (s, 1H), 1.66 1.54 (m, 1H), 1.53-1.48 (m, 1H), 0.94 (d, J=6.1Hz, 3H), 0.92 (d, J=6.1Hz, 3H)；¹³C NMR(100MHz,DMSO-d₆) δ 165.15,158.70 (d, J=236.9Hz, C-F), 149.56,138.07 (d, J=13.4Hz, C-CH-CF), 136.97,136.64,130.38,127.38,123.30 (d, J =8.5Hz, C-CH-CF), 110.84 (d, J=26.3Hz, CH-CF), 101.43 (d, J=28.2Hz, CH-CF), 52.48, 47.06,32.77,24.59,24.04,21.86；HRMS(ESI):m/z Calcd.For C₂₀H₂₄FN₄OS[M+H]⁺: 387.1655；Found:387.1649.

Embodiment 33. 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1-methyl isophthalic acid H-benzo [d] Imidazoles-4-Methanamide, the preparation of I-33:

The step 1. 7-tert-butyl group-2-(6-fluoro-4-(methoxycarbonyl)-1-methyl isophthalic acid H-benzo [d] imidazoles-2-base)-4,7-dihydro thiophene Fen also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the mode of operation of 6-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-7-isobutyl group -4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-13) (459mg, 0.94mmol) is prepared by raw material Obtaining white solid 292mg, productivity is 62% (petroleum ether: ethyl acetate=8:1).

Conformer 1:¹H NMR (300MHz, Chloroform-d) δ 7.72 (dd, J=10.3,2.5Hz, 1H), 7.37 (dd, J=11.5,2.5Hz, 1H), 7.23 (s, 1H), 5.22 (brs, 1H), 4.30 (dd, J=13.5,5.6Hz, 1H), 4.04 (s,3H),3.97(s,3H),3.40–3.14(m,2H),2.66–2.57(m,1H),1.48(s,9H),1.12(s,9H)；Structure As isomer 2:¹H NMR (300MHz, Chloroform-d) δ 7.72 (dd, J=10.3,2.5Hz, 1H), 7.37 (d, J= 11.5Hz, 1H), 7.23 (s, 1H), 5.02 (brs, 1H), 4.51 (dd, J=13.7,6.1Hz, 1H), 4.04 (s, 3H), 3.98 (s,3H),2.71–2.64(m,1H),2.66–2.57(m,1H),1.48(s,9H),1.12(s,9H)；MS(ESI)502 [M+H]⁺。

Step 2. 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1-methyl isophthalic acid H-benzo [d] miaow Azoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with the 7-tert-butyl group-2-(6-fluoro-4-(methoxycarbonyl)-1-methyl isophthalic acid H-benzene And [d] imidazoles-2-base)-4,7-dihydro-thiophene also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (6-4) (292mg, 0.58mmol) Obtaining white solid 196mg for raw material deprotection base, productivity is 84% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 7.72 (dd, J=10.2,2.4Hz, 1H), 7.35 (s, 1H), 7.23 (dd, J=7.95,2.4Hz, 1H), 4.05 (s, 3H), 3.98 (s, 3H), 3.86 (s, 1H), 3.37 (ddd, J=12.4,4.9, 2.8Hz, 1H), 2.91 (ddd, J=12.3,9.5,5.3Hz, 1H), 2.75-2.66 (m, 2H), 1.14 (s, 9H)；MS(ESI) 402[M+H]⁺。

Step 3. 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfide also [2,3-c] pyridine-2-base)-6-fluoro-1-methyl isophthalic acid H-benzo [d] miaow Azoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with 2-(the 7-tert-butyl group-4,5,6,7-Tetramethylene sulfides also [2,3-c] pyridine-2- Base)-6-fluoro-1-methyl isophthalic acid H-benzo [d] imidazoles-4-methyl formate (7-4) (196mg, 0.49mmol) is that raw material ammonia solves To white powder 132mg, productivity is 70% (dichloromethane: methanol=10:1).

¹H NMR(400MHz,TFA-d)δ8.20–7.52(m,3H),4.74(brs,1H),4.18(s,3H),4.06(brs, 1H),3.57(brs,1H),3.21(brs,2H),1.32(s,9H)；¹³C NMR(101MHz,TFA-d)δ167.72, 159.74,146.42,137.47,136.93,135.38,134.55,125.74,119.76,118.95,104.22,103.92,65.46, 42.79,35.06,32.31,24.56,21.62；HRMS(ESI):m/z Calcd.For C₂₀H₂₄FN₄OS[M+H]⁺: 387.1655；Found:387.1648.

The fluoro-2-of embodiment 34. 6-(4,5,6,7-oxolane also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

Step 1. 4,7-dihydrofuran also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

Reference literature prepares (list of references: Haginoya, N.；Kobayashi,S.；Komoriya,S.；Yoshino,T.； Suzuki,M.；Shimada,T.；Watanabe,K.；Hirokawa,Y.；Furugori,T.and Nagahara,T.Synthesis and Conformational Analysis of a Non-Amidine Factor Xa Inhibitor That Incorporates 5-Methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine as S4Binding Element.J.Med.Chem. 2004,47,5167-5182.)。

¹H NMR (300MHz, Chloroform-d) δ 7.39 (d, J=1.6Hz, 1H), 6.24 (d, J=1.6Hz, 1H), 4.44 (s, 2H), 3.63 (t, J=5.3Hz, 2H), 2.51 (t, J=5.3Hz, 2H), 1.48 (s, 9H)；MS(ESI)246 [M+Na]⁺。

Step 2. 2-formoxyl-4,7-dihydrofuran also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester:

According to preparing the operational approach of compound 2-1, with 4,7-dihydrofuran also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (14-13) (165mg, 0.74mmol) is that raw material prepares light yellow liquid 33mg, and productivity is 18% (oil Ether: ethyl acetate=10:1).

¹H NMR (300MHz, Chloroform-d) δ 9.55 (s, 1H), 7.09 (s, 1H), 4.54 (s, 2H), 3.67 (t, J= 5.2Hz, 2H), 2.60 (t, J=5.2Hz, 2H), 1.48 (s, 9H)；MS(ESI)274[M+Na]⁺。

Step 3. 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2-base)-4,7-dihydrofuran also [2,3-c] pyridine -6 (5H)-carboxylic acid tert-butyl esters:

According to preparing the operational approach of compound 3-1, with 2-formoxyl-4,7-dihydrofuran also [2,3-c] pyridine-6 (5H)-carboxylic acid The tert-butyl ester (2-14) (33mg, 0.13mmol) and 2, and 3-diaminourea-5-fluorophenyl carbamate (1-4) (24mg, 0.13 Mmol) being that raw material prepares faint yellow solid 46mg, productivity is 86% (petroleum ether: ethyl acetate=3:1).

¹H NMR (300MHz, Chloroform-d) δ 7.82 (dd, J=8.0,1.9Hz, 1H), 7.69 (dd, J=9.1,1.9 Hz, 1H), 7.56-7.46 (m, 2H), 4.57 (s, 2H), 4.06 (s, 3H), 3.768t, J=5.8Hz, 2H), 2.63 (t, J= 5.8Hz,2H),1.51(s,9H)；MS(ESI)416[M+H]⁺。

The fluoro-2-of step 4. 6-(4,5,6,7-oxolane also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-methyl formate:

According to preparing the operational approach of compound 4-1, with 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo [d] imidazoles-2- Base)-4,7-dihydrofuran also [2,3-c] pyridine-6 (5H)-carboxylic acid tert-butyl ester (3-18) (46mg, 0.11mmol) is that raw material goes Protection group obtains white solid 28mg, and productivity is 80% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 10.50 (brs, 1H), 7.61 (dd, J=7.6,2.4Hz, 1H), 7.59 ((dd, J=8.4,2.4Hz, 1H), 7.08 (s, 1H), 4.02 (s, 3H), 3.98 (s, 2H), 3.10 (t, J=5.6Hz, 2H), 2.59 (t, J=5.3Hz, 2H)；MS(ESI)316[M+H]⁺。

The fluoro-2-of step 5. 6-(4,5,6,7-oxolane also [2,3-c] pyridine-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

According to preparing the operational approach of compound I-1, with the fluoro-2-of 6-(4,5,6,7-oxolanes also [2,3-c] pyridine-2-base)-1H- Benzo [d] imidazoles-4-methyl formate (4-18) (28mg, 0.09mmol) is that raw material ammonia solution obtains pale powder 20mg, Productivity is 70% (dichloromethane: methanol=10:1).

¹H NMR(300MHz,DMSO-d₆) δ 9.09 (brs, 1H), 7.96 (brs, 1H), 7.56 (dd, J=10.7,2.5 Hz, 1H), 7.51 (dd, J=8.4,2.4Hz, 1H), 7.34 (s, 1H), 4.16 (s, 2H), 3.17 (t, J=5.0Hz, 2H), 2.68 (t, J=5.0Hz, 2H)；¹³C NMR(151MHz,DMSO-d₆) δ 172.01,165.47,158.78 (d, J= 237.0Hz, C-F), 157.99,149.26,145.61,143.50,118.56,113.29,110.51 (d, J=27.0Hz, CH-CF),102.11,49.05,42.21,41.57；HRMS(ESI):m/z Calcd.For C₁₅H₁₃FN₄O₂[M+H]⁺: 300.1023；Found:301.1093.

The fluoro-2-of embodiment 35. 6-(5-((methylamino) methyl) thiophene-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

Step 1. ((5-bromothiophene-3-base) methyl) (methyl) carbamate:

With 5-bromothiophene-3-formaldehyde as raw material, the intermediate that the method for reference literature obtains through reduction amination prepares mesh through protection Mark compound (list of references: Pyun, S.Y.；Lee,D.C.；Seung,Y.J.and Cho,B.R.Elimination Reactions of N-Alkyl-N-chlorothenylamines Promoted by MeONa-MeOH and Et2NH-MeCN. Effect of theβ-Aryl Group on the Imine-Forming Transition State.J.Org.Chem.2005,70, 5327-5330.)。

¹H NMR (300MHz, Chloroform-d) δ 6.87 (d, J=3.7Hz, 1H), 6.68 (d, J=3.7Hz, 1H), 4.44(s,2H),2.84(s,3H),1.49(s,9H)；MS(ESI)328[M+Na]⁺。

Step 2. ((5-formylthien-2-base) methyl) (methyl) t-butyl carbamate:

According to preparing the operational approach of compound 2-1, with ((5-bromothiophene-3-base) methyl) (methyl) t-butyl carbamate (12-14) (273mg, 0.89mmol) is that raw material prepares light yellow liquid 208mg, and productivity is 92% (stone Oil ether: ethyl acetate=10:1).

¹H NMR (300MHz, Chloroform-d) δ 9.86 (s, 1H), 7.64 (d, J=3.8Hz, 1H), 7.02 (d, J= 3.7Hz,1H),4.57(s,2H),2.89(s,3H),1.49(s,9H)；MS(ESI)278[M+Na]⁺。

Step 3. 2-(5-(((tertbutyloxycarbonyl) (methyl) amino) methyl) thiophene-2-base)-6-fluoro-1H-benzo [d] imidazoles-4-first Acid methyl ester:

According to preparing the operational approach of compound 3-1, with ((5-formylthien-2-base) methyl) (methyl) t-butyl carbamate (2-15) (208mg, 0.82mmol) and 2,3-diaminourea-5-fluorophenyl carbamate (1-4) (150mg, 0.82mmol) Preparing faint yellow solid 333mg for raw material, productivity is 97% (petroleum ether: ethyl acetate=3:1).

¹H NMR (300MHz, Chloroform-d) δ 10.44 (brs, 1H), 7.64 (dd, J=8.9,2.2Hz, 1H), 7.60 (dd, J=9.6,2.4Hz, 1H), 7.54 (d, J=3.7Hz, 1H), 7.00 (d, J=2.9Hz, 1H), 4.60 (s, 2H), 4.02 (s,3H),2.91(s,3H),1.50(s,9H)；MS(ESI)420[M+H]⁺。

The fluoro-2-of step 4. 6-(5-((methylamino) methyl) thiophene-2-base)-1H-benzo [d] imidazoles-4-carboxylate methyl ester:

According to preparing the operational approach of compound 4-1, with 2-(5-(((tertbutyloxycarbonyl) (methyl) amino) methyl) thiophene-2- Base)-6-fluoro-1H-benzo [d] imidazoles-4-methyl formate (3-19) (333mg, 0.79mmol) be raw material deprotection base obtain White solid 216mg, productivity is 86% (dichloromethane: methanol=50:1).

¹H NMR (300MHz, Chloroform-d) δ 7.60 (dd, J=15.6,2.4Hz, 1H), 7.57 (dd, J=16.2, 2.4Hz, 1H), 7.53 (d, J=3.4Hz, 1H), 6.97 (d, J=3.7Hz, 1H), 3.99 (brs, 5H), 2.50 (s, 3H)； MS(ESI)320[M+H]⁺。

The fluoro-2-of step 5. 6-(5-((methylamino) methyl) thiophene-2-base)-1H-benzo [d] imidazoles-4-Methanamide:

¹H NMR(300MHz,DMSO-d₆)δ9.00(brs,1H),7.90(brs,1H),7.79(brs,1H), (7.56-7.50 m, 2H), 7.08 (d, J=3.9Hz, 1H), 3.90 (s, 2H), 2.33 (s, 3H)；¹³C NMR(125MHz, DMSO-d₆) δ 165.09,158.17 (d, J=236.6Hz, C-F), 149.06,148.66,130.63,127.95,126.09, (122.22,109.76 d, J=26.2Hz, CH-CF), 101.94 (d, J=27.1Hz, CH-CF), 49.43,35.13；HRMS (ESI):m/z Calcd.For C₁₄H₁₄FN₄OS[M+H]⁺:305.0872；Found:305.0863.

Active testing is tested

Molecular level PARP1 inhibition of enzyme activity is tested

Experimental technique: enzyme linked immunosorbent assay (Enzyme-Linked Immunosorbent Assay, ELISA) (reference Document: Decker, P.；Miranda,E.A.；de Murcia,G.and Muller,S.An improved nonisotopic test to screen a large series of new inhibitor molecules of poly(ADP-ribose)polymerase activity for therapeutic applications.Clin.Cancer Res.1999,5,1169-1172.).Its principle is by substrate histone bag By on 96 orifice plates of adsorptivity, add PARP1 recombinase, substrate NAD⁺, activate DNA make PARP1 occur Enzyme reaction, makes histone generate product PAR (poly-adenosine diphosphate ribose), is subsequently adding anti-PAR (anti-PAR) Antibody, detect on 96 orifice plates the intensity of product PAR on coated histone, it is possible to reflect PARP1 enzyme Activity.

Concrete grammar is as follows:

1. histone is the important substrate that PARP1 generally acknowledges.Used PBS (the 10mM sodium phosphate buffer without potassium ion Liquid, 150mM NaCl, pH 7.2-7.4) it is coated 96 hole ELISA Plate, put 37 DEG C of incubator overnight；Discard liquid in hole.With The T-PBS (PBS containing 0.1%Tween-20) in 120 μ L/ holes washes plate 5 times.It is dried in 37 DEG C of baking ovens.

2. add NAD⁺(8 μMs/hole of final concentration), DNA (100ng/ hole), PARP1 (10ng/ hole) (use PARP1 Enzyme reaction buffer solution dilutes, buffer (trishydroxymethylaminomethane) Han 50mM Tris, 2mM MgCl₂, pH 8.0), The inhibitor that every hole adds certain diluted concentration (from the beginning of 10 μMs, 10 times dilute 6 gradients to final concentration) 10 μ L is (positive Control compound is: AZD2281, trade name Olaparib, purchased from LC, Laboratories company；ABT-888, business Name of an article Veliparib, purchased from Han Xiang bio tech ltd, Shanghai；2-(4-hydroxy phenyl)-1H-benzo [d] imidazoles-4-formyl Amine, numbered D-1, reference literature method prepares (list of references: White, A.W.；Almassy,R.；Calvert,A.H.； Curtin,N.J.；Griffin,R.J.；Hostomsky,Z.；Maegley,K.；Newell,D.R.；Srinivasan,S.and Golding,B.T.Resistance-modifying agents.9.Synthesis and biological properties of benzimidazole inhibitors of the DNA repair enzyme poly(ADP-ribose)polymerase.J.Med. Chem.2000,43,4084-4097.),¹H NMR(300MHz,Methanol-d₄) δ 8.03 (d, J=8.7Hz, 2H), 7.89 (dd, J=7.6,1.1Hz, 1H), 7.68 (dd, J=8.1,1.1Hz, 1H), 7.29 (dd, J=8.3,7.4Hz, 1H), 6.94 (d, J=8.7Hz, 2H)；MS(ESI)254[M+H]⁺.), each concentration arranges 2 repetitions, and reaction system is common 100 μ L/ holes (are supplied by aforementioned PARP1 enzyme reaction buffer solution), 37 DEG C of shaking tables reaction 1h, arrange blank, the positive, Negative control (with only add reaction buffer diluent as blank control wells, the enzyme-added hole of without inhibitor is Positive control wells, with Without inhibitor for negative control hole).Start reaction, put 37 DEG C of shaking tables and react 1 hour.

3. washing plate three times with PBS-T, (1:6000 uses to add anti-Anti-PAR Polyclonal Antibody (Rabbit) PBS-T dilution containing 5mg/mL BSA), 100 μ L/ holes, 37 DEG C of shaking tables hatch 1h；

4. washing plate three times with PBS-T, (1:2000, with containing 5 for two anti-(anti-mouse antibody) of addition peroxidase labelling The PBS-T dilution of μ g/mL BSA), 100 μ L/ holes, 37 DEG C of shaking tables react 30 minutes；

5. add OPD (O-Phenylenediamine Dihydrochloride) the nitrite ion 100 μ L/ hole of 2mg/mL (with containing 0.1%H₂O₂0.1M citric acid-sodium citrate buffer (pH=5.4) dilution), 25 DEG C of lucifuges are anti- Answer 15 minutes (need to be with ultrasonic when OPD dissolves, nitrite ion needs now with the current).

6. with the 2M H in 50 μ L/ holes₂SO₄Terminate reaction, by Molecular Devices microplate reader (purchased from U.S.'s molecule instrument Device company (MDC), model is SpectraMax 190Microplate Reader (90V to 240V)) reading, wavelength 490 Nm surveys OD value.

According to the below equation calculating medicine suppression degree to PARP1 enzymatic activity:

Suppression ratio (%)=(OD_{Control wells}-OD_{Dosing holes})/OD_{Control wells}× 100%

And calculate drug level when reaching 50% suppression ratio, i.e. IC by Logit method accordingly₅₀Value.Test in triplicate, Calculate meansigma methods and standard deviation.

Molecular level PARP2 inhibition of enzyme activity is tested

Experimentation is tested with molecular level PARP1 inhibition of enzyme activity, and institute is enzyme-added and substrate detailed data is: add NAD⁺ (8 μMs/hole of final concentration), DNA (100ng/ hole), PARP2 (10ng/ hole).

Body outer cell proliferation Inhibition test

Cell strain: chinese hamster fibroblast cell line strain VC8 (BRCA2^-/-)Wiegant,W.W.；Overmeer,R.M.； Godthelp,B.C.；van Buul,P.and Zdzienicka,M.Z.Chinese hamster cell mutant,V-C8,a Model for analysis of Brca2function.Mutation research 2006,600,79-88. and wild-type cell strain V79 (is given by Univ Leiden Netherlands Malgorzata professor Z.Zdzienicka).

Experimental technique: CCK8 (Cell Counting Kit-8) method.Containing WST 8: chemical name in CCK-8 reagent: 2-(2-methoxyl group-4-nitrobenzophenone)-3-(4-nitrobenzophenone)-5-(2,4-disulfonic acid benzene)-2H-tetrazolium monosodium salt, it is at electron carrier By the dehydrogenase in cell mitochondrial under the effect of 1-methoxyl group-5-toluphenazine dimethyl sulfate (1-Methoxy PMS) It is reduced to yellow first product (Formazan) with high water soluble.The quantity of the first thing generated and the number of living cells Amount is directly proportional.Cell proliferation is the most more fast, then color is the deepest；Cytotoxicity is the biggest, then color is the most shallow.For same thin Born of the same parents, the depth and the cell number of color are linear.At 450nm wavelength, measure its light with enzyme-linked immunosorbent assay instrument to inhale Receipts value, can indirectly reflect living cells quantity, thus calculate cell inhibitory rate.

Concrete grammar is as follows: will be in the cell of exponential phase by 2000/hole of V79 cell, 4000, VC8 cell The density in/hole is seeded to 96 well culture plates, every hole 100 μ L, and after overnight incubation, (final concentration is from 10M to add variable concentrations Start, 5 times dilution 6 gradients) medicine effect 72h, each concentration sets three wells, and sets the physiology salt of respective concentration Water Vehicle controls and acellular zeroing hole.After effect terminates, adding 10 μ L CCK8 reagent, 37 DEG C are continued to cultivate 4 hours, Microplate reader surveys OD450 value.

According to below equation calculating medicine cell proliferation suppression degree:

Experimental result (table 1) shows: following 6 compounds of the present invention are to the inhibitory activity of PARP1 and positive control AZD2281 is suitable, better than positive control ABT-888 activity, is I-4 (14.04nM) respectively, I-15 (16.87nM), I-22 (14.33nM), I-24 (15.06nM), I-28 (7.52nM), I-30 (18.47nM), I-32 (19.68 nM).And part of compounds shows certain selectivity to PARP1, such as I-4, I-5, I-6, I-15, I-16, I-18, I-21, I-22, be different from AZD-2281 and ABT-888.Body outer cell proliferation Inhibition test shows: the present invention Compound is to chinese hamster fibroblast cell line strain VC8 (BRCA2^-/-) there is good inhibitory activity, and intact to wild type The CHL cells strain VC79 (BRCA2 fallen into^+/+) unrestraint activity, show that designed compound is to BRCA defect Cell has the selectivity of height.

The molecular level of table 1. part benzimidazoles compound and cellular level biological activity result

Note: in table, "--" represents that maximum concentration is invalid or without result of calculation.

As can be known from Table 1, when with oxygen atom replacement sulphur atom (compound I-34 is to compound I-15), due to molecule The most there is not the n between sulphur atom and benzimidazole nitrogen-atoms₀σ * non-binding effect, causes compound I-34 to PARP1's Activity drastically declines.

Compared with compound I-35 with I-15, have employed open loop policy, the inhibitory activity of PARP1 has been declined by it, but The activity of PARP2 has been strengthened.

Compound I-30 is as follows to the growth inhibited effect of human breast carcinoma MDA-MB-436 nude mouse subcutaneous transplantation tumor:

Experimental technique

Animal: BALB/cA nude mouse, female, 4-5 week old, body weight 19 ± 2g, by Chinese Academy of Sciences's Shanghai drug research Being thered is provided, production licence is numbered: SCXK (Shanghai) 2013-001.The use quality certification is numbered: SYXK (Shanghai) 2013-0049. Every treated animal number: negative control group 12, administration group 6.Cell strain: human breast carcinoma MDA-MB-436 cell strain Being seeded to axillary fossa on the right side of nude mouse subcutaneous, cell inoculum concentration is 5 × 10⁶/ only, in nude mice, pass 2 after forming transplanted tumor again For rear use.

The tumor tissue taking growth animated period cuts into 1.5mm³Left and right, aseptically, is inoculated in axillary fossa on the right side of nude mouse Subcutaneous.Nude mouse subcutaneous transplantation tumor vernier caliper measurement transplanted tumor diameter, treats that tumor average volume grows to 200mm³ After Zuo You, by animal random packet.I-30 mono-hydrochloric salts 100mg/kg and 20mg/kg group, once a day oral administration, Successive administration 21 days.Positive control medicine AZD228130mg/kg, once a day oral administration, successive administration 21 days. Solvent control is to equivalent water for injection.

In whole experimentation, measure transplanted tumor diameter 2 times a week, weigh Mouse Weight simultaneously.

The computing formula of gross tumor volume (tumor volume, TV) is:

TV=1/2 × a × b²

Wherein a, b represent length and width respectively.

Result according to measuring calculates relative tumour volume (relative tumor volume, RTV), and computing formula is:

RTV=V_t/V₀

Wherein V₀For (d during sub-cage administration₀) measure gained gross tumor volume, V_tGross tumor volume during for measuring each time.

The evaluation index of anti-tumor activity is:

1) Relative tumor rate of increase T/C (%), computing formula is as follows:

T/C (%)=(T_RTV/C_RTV) × 100%

T_RTV: treatment group RTV；C_RTV: negative control group RTV；

2) gross tumor volume growth inhibition rate GI%, computing formula is as follows:

GI%=[1-(TVt-TV₀)/(CVt-CV₀)] × 100%

TVt is the tumor volume that treatment group is measured every time；TV₀Gained tumor volume when being administered for therapeutic component cage；CVt is right The tumor volume every time measured according to group；CV₀For gained tumor volume during matched group sub-cage administration；

3) tumor-like hyperplasia, computing formula is as follows:

Tumor-like hyperplasia %=(Wc-W_T)/Wc × 100%

Wc: matched group tumor weight, W_T: treatment group tumor weight.

Result is as shown in table 2, I-30 mono-hydrochloric salts oral administration every day 100mg/kg or 20mg/kg, and after one week, tumor is raw Length substantially slows down, and successive administration three Zhou Houneng significantly inhibits the growth of human breast carcinoma MDA-MB-436 Nude Mice, The T/C of the 21st day is 2.12% and 26.24%, and the effect of its suppression tumor growth is better than AZD228130mg/kg group. Table 2 shows, the body weight of nude mouse is affected less by I-30 mono-hydrochloric salts oral administration every day 100mg/kg or 20mg/kg.

The table 2. I-30 mono-hydrochloric salts Relative tumor rate of increase to human breast carcinoma MDA-MB-436 Nude Mice

T students test vs solvent control group, * p < 0.05**p < 0.001

The all documents mentioned in the present invention are incorporated as reference the most in this application, are individually recited just as each document As with reference to like that.In addition, it is to be understood that after the above-mentioned teachings having read the present invention, those skilled in the art are permissible Making various changes or modifications the present invention, these equivalent form of values fall within the model that the application appended claims is limited equally Enclose.

Claims

1. compound, R-isomer, S-isomer or its pharmaceutically acceptable salt as shown in formula I:

Wherein,

2. compound as claimed in claim 1, it is characterised in that R¹For hydrogen, the straight or branched alkyl of C1-C4, Or the cycloalkyl of C3-C6.

3. compound as claimed in claim 1, it is characterised in that R²For hydrogen, fluorine, chlorine, bromine, the straight chain of C1-C4 Or branched alkyl or the cycloalkyl of C3-C6.

4. compound as claimed in claim 1, it is characterised in that R⁴、R⁵Independently be substituted or unsubstituted C1-C4 Straight or branched alkyl, the cycloalkyl of C3-C6 or phenyl, described replacement refers to have 1-2 the substituent group selected from lower group: Hydroxyl, halogen, C1-C4 straight or branched alkoxyl.

5. compound as claimed in claim 1, it is characterised in that R⁶For hydrogen, the straight or branched alkyl of C1-C4 or The cycloalkyl of C3-C6.

6. compound as claimed in claim 1, it is characterised in that Y¹For substituted or unsubstituted methylene, Y²Replace Or unsubstituted ethylidene；Or Y¹For substituted or unsubstituted ethylidene, Y²For substituted or unsubstituted methylene, its In

7. compound as claimed in claim 1, it is characterised in that described compound is selected from lower group:

8. the preparation method of compound as claimed in claim 1, it is characterised in that described method includes described in formula II Compound generation ammonolysis obtain the step of the compound described in formula I:

Wherein, R¹、R²、Y¹、Y²、R³, X definition as claimed in claim 1.

9. a pharmaceutical composition, it is characterised in that described pharmaceutical composition include the compound described in claim 1, R-isomer, S-isomer or its pharmaceutically acceptable salt；And

Pharmaceutically acceptable carrier.

10. compound as claimed in claim 1 or the purposes of pharmaceutical composition as claimed in claim 9, its feature exists In, it is used for preparing: (1) PARP1 inhibitor；(2) PARP2 inhibitor；(3) prevent and/or treat the medicine of tumor； (4) anti-inflammatory drug；And/or (5) prevent and/or treat and the medicine of PARP relevant disease.