CN109734700A - SMO inhibitor of the piperidine structure containing benzoyl and preparation method thereof and purposes - Google Patents

Abstract

The invention discloses a kind of benzoyl piperidine derivatives and its preparation method and application; the benzoyl piperidine derivatives are a kind of compounds for containing N- methyl-N- (1- (2- amino benzoyl) piperidin-4-yl) amide structure, and structure is shown in formula I.The invention also discloses the preparation methods of the benzoyl piperidine derivatives.The invention also discloses the benzoyl piperidine derivatives to prepare the application in SMO protein inhibitor, and purposes of the pharmaceutical composition containing these compounds in terms of preparing anti-tumor drug.

Description

SMO inhibitor of the piperidine structure containing benzoyl and preparation method thereof and purposes

Technical field

The invention belongs to field of pharmaceutical chemistry technology, and in particular to one kind contains N- methyl-N- (1- (2- aminobenzoyl Base) piperidin-4-yl) amide structure SMO protein inhibitor, their preparation method, and the drug containing these compounds Composition and its purposes in terms of preparing anti-tumor drug.

Technical background

Tumour is to endanger the major disease of human health.With rising year by year for morbidity and mortality, malignant tumour is Through becoming the global primary cause of death and important public health problem.Chemotherapy is the main means of clinical treatment tumour One of, therefore the research of anti-tumor drug is always the emphasis and hot spot of pharmaceuticals industry research.

Stem cell is biomedical research frontier in the ascendant in recent years, has great clinical value.Into It since entering 21 century, has been constantly improve as tumor stem cell is theoretical, researcher has for why tumour is difficult to thoroughly cure New understanding.A large amount of evidences show that tumor stem cell present in tumor tissues is insensitive to existing chemotherapy and radiotherapy measure, And this part cell has the ability for being differentiated to form tumour cell again, therefore, can target killing tumor stem cell will become Thoroughly cure the key factor of cancer.

It is quite similar in view of the fundamental characteristics of tumor stem cell and adult stem cell, then the key of targeting somatocyte development Access can become the means of killing tumor stem cell.At present for access Notach, Hedgehog relevant to development and The research of Wnt inhibitor has been achieved for a series of achievements, wherein for the inhibitor of key protein SMO in Hedgehog access By FDA approval listing for treating basal-cell carcinoma.However, clinical research discovery is mutated due to SMO albumen (D473H), suffer from Person produces drug resistance to the drug listed quickly.Therefore, exploitation is directed to the second generation SMO inhibitor of SMO mutant drug-resistant, gives Ongoing research area brings new opportunities and challenges.

Summary of the invention

Goal of the invention: the present invention provides one kind to contain N- methyl-N- (1- (2- amino benzoyl) piperidin-4-yl) acyl The benzoyl piperidine analog derivative of amine structure, and the specific preparation method and application of the derivative are provided in preparation SMO egg The pharmacy application of white inhibitor.

Technical solution: the invention discloses a kind of benzoyl piperidine analog derivative as shown in general formula I or its pharmaceutically Acceptable salt:

Wherein:

R¹Selected from substituted or non-substituted phenyl, wherein substituted phenyl be by one, it is two or more chosen from the followings The phenyl that substituent group replaces: halogen ,-NO₂、-NH₂,-CN, C1-C6 alkoxy ,-CHO ,-CF₃、-CONH₂, C1-C6 alkanoyl ,- SO₂CH₃、-NHCOCH₃、-NHCOCH₂CH₃、-CH₂CONH₂Or-NHCONH₂；

R²Selected from substituted or non-substituted five yuan or hexa-atomic aromatic heterocyclic or substituted or non-substituted C6-C12 aromatic hydrocarbons Base, wherein substituted five yuan or hexa-atomic aromatic heterocyclic are five yuan replaced by one, two or more substituent groups chosen from the followings Or hexa-atomic aromatic heterocyclic: halogen, C1-C6 alkyl ,-CN ,-NH₂、-NHCOCH₃, C1-C6 alkanoyl, C1-C6 alkoxy or- COOH；Substituted C6-C12 aryl is the C6-C12 aryl replaced by one, two or more substituent groups chosen from the followings: Halogen, C1-C6 alkyl ,-CN ,-NH₂、-NHCOCH₃, C1-C6 alkanoyl, C1-C6 alkoxy or-COOH.

As preferred:

R¹It is selected fromWherein R³And R⁴It is each independently selected from-H, halogen ,-NO₂、-NH₂、-CN、-OCH₃、- CHO、-CF₃、-CONH₂、-COCH₃、-SO₂CH₃、-NHCOCH₃、-NHCOCH₂CH₃、-CH₂CONH₂Or-NHCONH₂；

R²Selected from substituted or non-substituted five yuan or hexa-atomic aromatic heterocyclic containing N or substituted or non-substituted phenyl or connection Phenyl, wherein substituted five yuan or hexa-atomic aromatic heterocyclic containing N are replaced by one, two or more substituent groups chosen from the followings Five yuan or hexa-atomic aromatic heterocyclic containing N: halogen ,-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、-CN、-NH₂、-NHCOCH₃、-COCH₃、- COCH₂CH₃、-COCH₂CH₂CH₃、-OCH₃、-OCH₂CH₃、-OCH₂CH₂CH₃Or-COOH；Substituted phenyl or xenyl is by one The phenyl or xenyl that a, two or more substituent groups chosen from the followings replace: halogen ,-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、- CN、-NH₂、-NHCOCH₃、-COCH₃、-COCH₂CH₃、-COCH₂CH₂CH₃、-OCH₃、-OCH₂CH₃、-OCH₂CH₂CH₃Or-COOH.

It is further preferred:

R¹It is selected fromWherein R³Selected from-H, halogen ,-NO₂、-NH₂、-CN、-OCH₃、-CHO、-CF₃、- CONH₂、-COCH₃、-SO₂CH₃、-NHCOCH₃、-NHCOCH₂CH₃、-CH₂CONH₂Or-NHCONH₂；R⁴Selected from-H or-CF₃；

R²Selected from substituted or non-substituted five yuan or hexa-atomic aromatic heterocyclic containing N or substituted or non-substituted phenyl or connection Phenyl, wherein substituted five yuan or hexa-atomic aromatic heterocyclic containing N are replaced by one, two or more substituent groups chosen from the followings Five yuan or hexa-atomic aromatic heterocyclic containing N: halogen ,-CH₃、-CN、-NH₂、-NHCOCH₃、-COCH₃、-OCH₃Or-COOH；Replace Phenyl or xenyl are the phenyl or xenyl replaced by one, two or more substituent groups chosen from the followings: halogen ,-CH₃、- CN、-NH₂、-NHCOCH₃、-COCH₃、-OCH₃Or-COOH.

More preferably:

R¹It is selected from

R²It is selected from

Above preferred or further preferred or preferred group, it is preferred simultaneously not necessarily to represent these groups, Such as preferably in, R¹It can be used as only preferred, R²Can not preferably, certain R¹And R²It can also simultaneously preferably.

Term " aryl " refers to arene compounds group, such as phenyl, xenyl, naphthalene, anthryl, phenanthryl etc..

Term " aromatic heterocyclic " refers to containing the heteroatomic heteroaromatic class compound group such as N, S or O, such as phenanthridinyl, Carbazole, dibenzofurans, pyrrole radicals, triazolyl, pyridyl group, pyrazinyl, indyl, pyrimidine radicals or quinolyl etc..

Term " C1-C6 alkyl " refer to carbon atom number be 1 to 6 linear or branched alkyl group, such as methyl, ethyl, third Base, isopropyl, normal-butyl, isobutyl group, sec-butyl, tert-butyl etc..

Term " C1-C6 alkanoyl " refers to that carbon atom number is 1 to 6 linear or branched alkyl group acyl group, such as C1 alkane acyl Base refers to that acetyl group, C2 alkanoyl refer to propiono.

Term " C1-C6 alkoxy " refers to that carbon atom number is 1 to 6 straight or branched alkoxyl, such as methoxyl group, second Oxygroup, propoxyl group, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy etc..

Term " substitution " refers to monosubstituted or polysubstituted.

Preferably, herein described benzoyl piperidine derivatives are selected from I-1 to I-17:

The preparation method of the benzoyl piperidine analog derivative, including by compound IV and IX through condensation reaction preparationization Close object I:

Preferably, the preparation method of the benzoyl piperidine analog derivative, comprising the following steps:

(1) compound II reacts preparation intermediate compound IV through Buchwald-Hartwig with III；

(2) compound V prepares VI through reductive amination process；Again by compound VI and VII through condensation reaction prepare compound VIII；Protecting group prepare compound IX is sloughed by compound VIII again；

(3) compound IX and intermediate compound IV are through condensation reaction prepare compound I:

Alternatively, the preparation method of the benzoyl piperidine analog derivative, including passed through by compound XI and XII Buchwald-Har twig reacts prepare compound I:

Wherein, A is halogen.

It is preferred that including by compound IX-1 and X through condensation reaction prepare compound XI；It is passed through again by compound XI and XII Buchwald-Hartwig reacts prepare compound I.

Wherein, A is halogen.

For example, can be prepared by following method:

The preparation method one of compound of Formula I:

Wherein R¹It representsWherein R^1`Representative-NO₂、-OCH₃、-F、-CN、-CF₃、- COCH₃Or-SO₂CH₃。

Further, the process by compound IV and IX through condensation reaction prepare compound I: condensing agent used is selected from hexafluoro Phosphoric acid benzotriazole -1- base-oxygroup tripyrrole alkyl phosphorus (PyBOP), 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide salt Hydrochlorate (EDCI), dicyclohexylcarbodiimide (DCC) or N, N'- carbonyl dimidazoles (CDI), preferably EDCI；Solvent for use is selected from Methylene chloride, n,N-Dimethylformamide (DMF), n,N-dimethylacetamide (DMAC) or dimethyl sulfoxide (DMSO), preferably two Chloromethanes；Reaction temperature is selected from 0 DEG C~50 DEG C, preferably 20 DEG C~30 DEG C；Reaction time is 4h~48h, preferably 8h~16h.Institute State compound IV: compound IX: condensing agent (molar ratio) proportion is 1:1:1~1:10:10, preferably 1:1:1~1:2:2.

The preparation method two of compound of Formula I:

Wherein R²It representsWherein R^2`Representative-H ,-CN ,-OCH₃Or-COOH.

Further, the process of prepare compound I is reacted through Buchwald-Hartwig with XII by compound XI: described Compound XII be aniline, 4- cyano-aniline, 2- cyano-aniline, 4- aminoanisole, 2- aminoanisole, 4-aminopyridine, 2-aminopyridine, 4- phenylaniline, 4-aminobenzoic acid；Solvent for use is selected from toluene, dimethylbenzene, tetrahydrofuran (THF), second two Diethylene glycol dimethyl ether (DME), dioxane, N,N-dimethylformamide (DMF), N-Methyl pyrrolidone (NMP), dimethyl sulfoxide (DMSO) or in which the mixed solvent of any two or three of solvent, preferred toluene；Used catalyst is selected from three (dibenzylidenes third Ketone) two palladium (Pd₂(dba)₃) or palladium acetate (Pd (OAc)₂), preferably Pd (OAc)₂；Ligand used be selected from 2,2'- it is bis--(diphenyl phosphine Base) -1,1'- dinaphthalene (BINAP), three (2- tolyl) phosphine (P (o-tolyl)₃) or bis- (the diphenylphosphine) -9,9- dimethyl of 4,5- Xanthene (Xantphos), preferably BINAP；Reaction temperature is selected from 70 DEG C~110 DEG C, preferably 100 DEG C~110 DEG C；Reaction time For 4h~48h, preferably 8h~16h.The compound XI: compound XII: catalyst: ligand (molar ratio) proportion is 1:1: 0.01:1~1:10:0.5:5, preferably 1:1:0.1:2~1:2:0.2:3.

The preparation method of key intermediate IV in compound of Formula I preparation method of the present invention, comprising:

Further, the process of prepare compound IV is reacted through Buchwald-Hartwig with III by compound II: used Catalyst is selected from tris(dibenzylideneacetone) dipalladium (Pd₂(dba)₃), palladium acetate (Pd (OAc)₂) or cuprous iodide (CuI), preferably CuI；Alkali used is selected from sodium tert-butoxide (NaOtBu), hexamethyldisilazide lithium (LiHMDS), potassium carbonate (K₂CO₃), potassium phosphate (K₃PO₄) or cesium carbonate (Cs₂CO₃), preferred potassium carbonate；Solvent for use is selected from toluene, dimethylbenzene, tetrahydrofuran (THF), second two Diethylene glycol dimethyl ether (DME), dioxane, N,N-dimethylformamide (DMF), N-Methyl pyrrolidone (NMP), dimethyl sulfoxide (DMSO) or in which the mixed solvent of any two or three of solvent, preferably DMF；Reaction temperature is selected from 70 DEG C~110 DEG C, preferably 100 DEG C~105 DEG C；Reaction time is 4h~48h, preferably 8h~16h.The compound II: compound III: catalyst: alkali (molar ratio) proportion is 1:1:0.1:1~1:10:1:10, preferably 1:1:0.2:2~1:1.5:0.5:5.

The preparation method of key intermediate IX in compound of Formula I preparation method of the present invention, comprising:

Wherein R¹It representsWherein R^1`Representative-NO₂、-OCH₃、-F、-CN、-CF₃、- COCH₃Or-SO₂CH₃。

Further, the process by compound V through reductive amination process prepare compound VI: reducing agent used is selected from boron hydrogen Change sodium (NaBH₄), sodium triacetoxy borohydride (NaBH (OAc)₃) or sodium cyanoborohydride (NaBH₃CN), preferred cyano boron hydrogen Change sodium；Solvent for use is selected from ethyl acetate, methanol, tetrahydrofuran, chloroform, acetonitrile, methylene chloride, toluene or in which two kinds any Or the mixed solvent of three kinds of solvents, preferred methanol；Reaction temperature is selected from 0 DEG C~50 DEG C, preferably 20 DEG C~30 DEG C；Reaction time is 4h~for 24 hours, preferably 8h~16h.The compound V: methylamine: reducing agent (molar ratio) proportion be 1:1:1~1:10:5, preferably 1: 1.5:1.5~1:3:3.

Process by compound VI and VII through condensation reaction prepare compound VIII: the compound VII is 2- trifluoro Methyl -4- fluobenzoic acid, 4- nitrobenzoic acid, 4- methoxy benzoic acid, 4- fluobenzoic acid, 4- cyanobenzoic acid, 4- fluoroform Yl benzoic acid, 4- formylbenzoate, 4- thiamphenicol benzoic acid；Chlorination reagent used is selected from thionyl chloride (SOCl₂), trichlorine oxygen Phosphorus (POCl₃), phosphorus pentachloride (PCl₅) or oxalyl chloride ((COCl)₂), preferred oxalyl chloride；Acid binding agent used is selected from triethylamine, N, N- Diisopropylethylamine (DIEA), 4-dimethylaminopyridine (DMAP), pyridine, sodium acetate, sodium carbonate or potassium carbonate, preferably three second Amine；Solvent for use is selected from methanol, ethyl alcohol, methylene chloride, acetone, ethyl acetate, tetrahydrofuran or in which two or three any The mixed solvent of solvent, preferably methylene chloride；Reaction temperature is selected from 0 DEG C~50 DEG C, preferably 0 DEG C~30 DEG C；Reaction time is 0.5h~5h, preferably 1h~2h.The compound VI: compound VII: chlorination reagent: acid binding agent (molar ratio) proportion is 1:1: 1:2~5:1:5:10, preferably 1:1:1:2~2:1:2:4.

The process of protecting group prepare compound IX is sloughed by compound VIII: deprotecting regent used be selected from hydrogen chloride, Hydrogen bromide, sulfuric acid or trifluoroacetic acid, preferably trifluoroacetic acid；Solvent is methanol, ethyl alcohol, methylene chloride, acetone, ethyl acetate, four The mixed solvent of hydrogen furans or any the two, preferably methylene chloride；Reaction temperature is selected from 0 DEG C~50 DEG C, preferably 20 DEG C~30 DEG C； Reaction time is 0.5h~5h, preferably 1h~2h.

The preparation method of key intermediate XI in compound of Formula I preparation method of the present invention, comprising:

Further, the process by compound X and IX-1 through condensation reaction prepare compound XI: chlorination reagent used is selected from Thionyl chloride (SOCl₂), phosphorus oxychloride (POCl₃), phosphorus pentachloride (PCl₅) or oxalyl chloride ((COCl)₂), preferred oxalyl chloride；Institute Triethylamine, N, N- diisopropylethylamine (DIEA), 4-dimethylaminopyridine (DMAP), pyridine, sodium acetate, carbon are selected from acid binding agent Sour sodium or potassium carbonate, preferably triethylamine；Solvent for use is selected from methanol, ethyl alcohol, methylene chloride, acetone, ethyl acetate, tetrahydrofuran Or in which the mixed solvent of any two or three of solvent, preferred methylene chloride；Reaction temperature is selected from 0 DEG C~50 DEG C, preferably 0 DEG C ~30 DEG C；Reaction time is 1h~10h, preferably 3h~6h.The compound X: compound IX-1: chlorination reagent: acid binding agent (rubs That ratio) it matches as 1:1:1:1~5:1:5:5, preferably 1:1:1:1~2:1:2:2.

The invention also discloses a kind of pharmaceutical composition, containing above-mentioned compound of Formula I or its is pharmaceutically acceptable Salt and pharmaceutically acceptable auxiliary material.The compound can add pharmaceutically acceptable auxiliary material and common medicinal system is made The common medicines such as fragrance, sweetener, liquid or solid filler or diluent can be added such as tablet, capsule, syrup, suspending agent in agent Use auxiliary material.

The benzoyl piperidine derivatives or its pharmaceutically acceptable salt of general formula I of the present invention are in preparation SMO egg Application in white inhibitor medicaments.

The benzoyl piperidine derivatives and its stereoisomer of general formula I of the present invention, hydrate, solvate or Crystallize prepare in SMO protein inhibitor drug using also within the scope of the present invention.

The benzoyl piperidine derivatives or its pharmaceutically acceptable salt of general formula I of the present invention prepare it is antitumor Application in drug is also within the scope of the present invention.

Further, wherein SMO protein inhibitor is used to prepare the drug for treating malignant tumour.

The utility model has the advantages that the invention discloses benzoyl piperidine derivatives shown in a kind of new general formula I, pharmacological evaluation is aobvious Show, the compound of the present invention I can generate good inhibiting effect to SMO albumen, can be used for preparing treatment Hedgehog access The drug of the malignant tumour of overactivity；The invention also discloses the preparation methods of the benzoyl piperidine derivatives.

Specific embodiment

The application is explained in detail combined with specific embodiments below.

Embodiment 1

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -2- trifluoro The synthesis of methyl -4- fluorobenzamide (I-1)

The synthesis of 2- ((1- methyl-1 H- pyrazoles -5- base) amino)-benzoic acid (IV)

By o-iodobenzoic acid (II) (5.00g, 20.00mmol), 1- methyl-1-H-5- amino-pyrazol (III) (1.90g, 20.00mmol), potassium carbonate (5.50g, 40.00mmol) and CuI (0.76g, 4.00mmol) are dissolved in DMF (20.00ml), temperature control In 100~105 DEG C of stirring 8h.TLC (petroleum ether: ethyl acetate=1:1) detection raw material II has been reacted, and is stopped heating, is cooled to Room temperature.Reaction solution is poured into water (60.00ml), uses 1.00mol/L HCl to adjust reaction system pH=5 after stirring 20min ~6, brown solid is precipitated, filters, drying obtains brown solid 3.10g, yield 70%, m.p.134~139 DEG C.

¹H-NMR(300MHz,CDCl₃)δ(ppm):10.77-8.78(m,2H,NH,ArH),8.14-7.30(m,3H, ArH),6.89(s,1H,ArH),6.46(s,1H,ArH),3.85(s,3H,CH₃ ).

The synthesis of 4- (methylamino) piperidines -1- carboxylate (VI)

N-Boc- piperidin-4-one (V) (1.00g, 5.00mmol) and methylamine alcohol solution (1.00ml) are dissolved in methanol (10.00ml) adjusts pH=5~6 with acetic acid, 1h is stirred at room temperature.Be added portionwise under ice bath sodium cyanoborohydride (0.48g, 7.60mmol), 12h is reacted at room temperature.TLC (methylene chloride: methanol=10:1) detection raw material V has reacted.It is spin-dried for methanol, with saturation NaOH solution adjusts pH=9, is extracted with methylene chloride (10.00ml x 3), and anhydrous sodium sulfate dries, filters, is spin-dried for solvent and obtains Light yellow oil 1.00g, yield 100%.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 3.98 (d, J=10.20Hz, 2H, Boc-NCH₂ ), 2.74 (t, J= 12.20Hz,2H,BocNCH₂ ),2.48-2.43(m,1H,NHCH),2.38(s,3H,NHCH₃ ), 1.79 (d, J=12.20Hz, 2H,NHCHCH₂ ), 1.39 (s, 9H, Boc-H), 1.17 (dd, J=20.20,10.20Hz, 2H, NHCHCH₂ ).

The conjunction of 4- (fluoro- N- methyl -2- (trifluoromethyl) benzamido of 4-) piperidines -1- carboxylate (VIII-1) At

2- trifluoromethyl -4- fluobenzoic acid (VII-1) (3.00g, 14.40mmol) is dissolved in methylene chloride (5.00ml), It is cooled to 0 DEG C after 2 drop DMF are added, is instilled oxalyl chloride (1.50ml, 17.30mmol), 30min is stirred at room temperature in dislocation, is spin-dried for dichloro It is spare that methane obtains acyl chlorides.

N-Boc-4- methylamino piperidine (VI) (3.70g, 17.30mmol) is dissolved in methylene chloride (20.00ml), is added Triethylamine (3.00ml, 21.60mmol), is cooled to 0 DEG C, and 1h is stirred at room temperature in dislocation after instilling the dichloromethane solution of acyl chlorides.TLC (petroleum ether: ethyl acetate=3:1) detection has been reacted.1.00mol/L HCl, saturated sodium bicarbonate aqueous solution, saturation are used respectively Brine It reaction solution, anhydrous sodium sulfate is dry, through column chromatography (petroleum ether: ethyl acetate=6:1) separation, obtains colorless oil Object 5.33g, yield 91.30%.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.44-7.39(m,1H,ArH),7.32-7.29 (m,2H,ArH),4.73(m,1H,CH₃NCH),4.25(m,2H,BocNCH₂ ),2.88-2.82(m,2H,BocNCH₂ ),2.63 (s,3H,NCH₃ ),1.79-1.63(m,4H,CH₃N(CH₂)₂ ),1.46(s,9H,Boc-H).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.44-7.39(m,1H,ArH),7.32-7.29 (m,2H,ArH),4.15-4.11(m,2H,BocNCH₂ ),3.23(s,1H,CH₃NCH),2.98(s,3H,NCH₃ ),2.51-2.35 (m,2H,BocNCH₂ ),1.79-1.63(m,4H,CH₃N(CH₂)₂ ),1.43(s,9H,Boc-H).

The synthesis of fluoro- N- methyl-N- (the piperidin-4-yl) -2- trifluoromethyl benzamide (IX-1) of 4-

Compound VIII-1 (4.33g, 10.70mmol) is dissolved in methylene chloride (20.00ml), trifluoro second is added at room temperature Sour (5.00ml), is stirred at room temperature 1h.TLC (methylene chloride: methanol=10:1) detection raw material has reacted, and is spin-dried for solvent, is added Water (15.00ml) adjusts pH=9 with saturation NaOH solution, is extracted with methylene chloride (15.00ml x 3), anhydrous sodium sulfate is dry It is dry, it is spin-dried for obtaining light butter object 3.20g, yield 98%.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.46-7.41(m,1H,ArH),7.33-7.30 (m,2H,ArH),5.66(br,1H,NH),4.78(m,1H,CH₃NCH),3.44-3.40(m,2H,HNCH₂ ),2.98-2.90(m, 2H,BocNCH₂ ),2.68(s,3H,NCH₃ ),2.09-1.58(m,4H,CH₃N(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.46-7.41(m,1H,ArH),7.33-7.30 (m,2H,ArH),5.66(brs,1H,NH),3.27-3.13(m,3H,CH₃NCHCH₂CH₂ NH),3.02(s,3H,NCH₃ ), 2.51-2.37(m,2H,BocNCH₂ ),2.09-1.58(m,4H,CH₃N(CH₂)₂ ).

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -2- trifluoro The synthesis of methyl -4- fluorobenzamide (I-1)

Intermediate compound IV (500.00mg, 2.30mmol) and intermediate compound I X-1 (699.00mg, 2.30mmol) are dissolved in dichloro Methane (20.00ml) is added HOBt (372.00mg, 2.76mmol), then EDCI (530.00mg, 2.76mmol) is added portionwise, 12h is stirred at room temperature.TLC (methylene chloride: methanol=25:1) detection raw material IX-1 has reacted, and stops reaction, successively uses 1.00mol/L HCl (10.00mlx2), saturated sodium bicarbonate (10.00ml x 2), saturated salt solution (10.00ml x 2) are washed Wash, anhydrous sodium sulfate dries, filters, and filtrate decompression is concentrated to dryness, residue through column chromatograph (methylene chloride: methanol=200:1~ It 100:1) separates, obtains dark green solid 0.74g, yield 64.00%, m.p.206~207 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.47-7.41(m,2H,ArH),7.34-7.29 (m,3H,ArH),7.27-7.12(m,2H,ArH,NH),6.91-6.75(m,2H,ArH),6.06(s,1H,ArH),5.00- 4.81(m,1H,CH₃NCH),4.80-4.12(m,2H,CON(CH₂)₂ ),3.72(s,3H,CH₃),3.29-3.03(m,2H,CON (CH₂)₂ ),2.67(s,3H,CH₃),1.84(m,4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.47-7.41(m,2H,ArH)7.34-7.29 (m,3H,ArH),7.27-7.12(m,2H,ArH),6.91-6.75(m,2H,ArH),6.06(s,1H,ArH),4.80-4.12 (m,2H,CON(CH₂)₂ ),3.70(s,3H,CH₃),3.43-3.60(m,1H,CH₃NCH),3.02(s,3H,CH₃),2.64-2.55 (m,2H,CON(CH₂)₂ ),1.84(m,4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 169.54,167.52,161.65 (d, J= 251.50Hz),143.59,138.09,130.89,128.91,128.80,128.50,128.40,127.68,122.30(q,J =276.20Hz), 119.18,118.65 (d, J=36.10Hz), 118.47,114.64,113.95 (ddd, J=25.30, 9.20,4.70Hz),98.11,50.54,34.46,31.11,28.64,27.95.

Minor rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 169.54,167.52,161.65 (d, J= 251.50Hz),143.59,138.09,131.02,128.91,128.80,128.50,128.40,127.54,122.30(q,J =276.20Hz), 119.18,118.65 (d, J=36.10Hz), 118.47,114.64,113.95 (ddd, J=25.30, 9.20,4.70Hz),98.25,56.13,34.46,29.41,28.99,27.02.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₆F₄N₅O₂:504.2023；found 504.2008.

IR(cm^-1):3345.86,1630.91,1589.00,1559.42,1503.80,1467.52,1445.43, 1422.09,1367.50,1330.25,1319.31,1271.16,1210.59,1171.33,1074.76,1040.48, 1008.04,904.29,859.78,760.33,659.16,606.26.

Embodiment 2

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- nitro The synthesis of benzamide (I-2)

The synthesis of 4- (N- methyl -4- nitrobenzoyl acylamino-) piperidines -1- carboxylate (VIII-2)

With 4- nitrobenzoic acid (VII-2) (2.40g, 14.40mmol) for raw material, same VIII-1 is operated, chromatographs (stone through column Oily ether: ethyl acetate=6:1) separation, faint yellow solid 4.30g is obtained, yield: 82%, m.p.152~153 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 8.31 (d, J=7.90Hz, 2H, ArH), 7.58 (d, J=7.20Hz, 2H,ArH),4.72(s,1H,CONCH),4.26(brs,2H,CONCHCH₂ ),3.01(brs,2H,CONCHCH₂ ),1.77(s, 3H,CONCH₃ ),1.49(brs,4H,BocN(CH₂)₂ ),1.49(s,9H,Boc-H).

The synthesis of N- methyl -4- nitro-N- (piperidin-4-yl) benzamide (IX-2)

With VIII-2 (4.89g, 10.70mmol) for raw material, same IX-1 is operated, obtains light yellow oil 2.70g, yield: 97%.N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- nitrobenzoyl The synthesis of amide (I-2)

With IV (500.00mg, 2.30mmol), IX-2 (605.00mg, 2.30mmol) is raw material, same I-1 is operated, through column (methylene chloride: methanol=200:1~100:1) separation is chromatographed, obtains faint yellow solid 659.00mg, yield: 62%, m.p.91~ 92℃。

¹H-NMR(300MHz,CDCl₃) δ (ppm): 8.31 (d, J=8.40Hz, 2H, ArH), 7.58 (d, J=8.40Hz, 2H,ArH),7.47(s,1H,ArH),7.30-7.16(m,3H,ArH,NH),6.90-6.81(m,2H,ArH),6.04(s,1H, ArH),4.85(s,1H,CH₃NCH),4.69-4.30(m,2H,CONCH₂ ),3.73(s,3H,CH₃),3.14-3.01(m,2H, CONCH₂ ),2.84(s,3H,CH₃),1.89-1.74(m,4H,CONCH(CH₂)₂ ).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):170.06,169.34,148.38,144.18,142.61,138.58, 131.44,128.11,127.87,127.74,127.67,123.95,118.91,115.14,98.42,51.62,34.94, 32.08,29.97,29.02.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₇N₆O₄:463.2094；found 463.2081.

IR(cm^-1):3243.39,2934.42,1625.07,1557.39,1522.37,1445.61,1351.24, 1328.59,1274.16,1070.27,1016.01,924.94,856.70,759.52,698.26.

Embodiment 3

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) 4- methoxyl group The synthesis of benzamide (I-3)

The synthesis of 4- (4- methoxy-. N-methyl benzamido) piperidines -1- carboxylate (VIII-3)

With 4- methoxy benzoic acid (VII-3) (2.20g, 14.40mmol) for raw material, same VIII-1 is operated, is chromatographed through column (petroleum ether: ethyl acetate=6:1) separation, obtains white solid 4.40g, yield: 88%, m.p.116~117 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 7.39 (d, J=8.50Hz, 2H, ArH), 6.94 (d, J=8.60Hz, 2H,ArH),4.62(brs,1H,CONCH),4.25-4.21(m,2H,CONCHCH₂ ),3.87(s,3H,OCH₃),2.91(s,3H, NCH₃),2.77(brs,1H,ONCHCH₂ ),1.73(brs,4H,BocN(CH₂)₂ ),1.51(s,9H,Boc-H).

The synthesis of 4- methoxy-. N-methyl-N- (piperidin-4-yl) benzamide (IX-3)

With VIII-3 (3.70g, 10.70mmol) for raw material, same IX-1 is operated, obtains colorless oil 2.56g, yield: 96.30%.N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) 4- methoxyl group The synthesis of benzamide (I-3)

With IV (500.00mg, 2.30mmol), IX-3 (570.00mg, 2.30mmol) is raw material, same I-1 is operated, through column (methylene chloride: methanol=200:1~100:1) separation is chromatographed, obtains faint yellow solid 740.00mg, yield: 72%, m.p.162 ~163 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 7.47 (d, J=1.80Hz, 1H, ArH), 7.38 (d, J=8.70Hz, 2H, ArH), 7.29 (s, 1H, NH), 7.24-7.21 (m, 2H, ArH), 6.94 (d, J=8.70Hz, 2H, ArH), 6.87 (t, J= 7.40Hz, 1H, ArH), 6.81 (d, J=8.20Hz, 1H, ArH), 6.05 (d, J=1.70Hz, 1H, ArH), 4.76-4.32 (m, 3H,NCH₃CH,CON(CH₂)₂ ),3.86(s,3H,OCH₃),3.73(s,3H,CH₃),3.16-2.96(m,2H,CON(CH₂)₂ ), 2.91(s,3H,CH₃),1.93-1.74(m,4H,NCH3CH(CH₂)₂ ).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):171.63,169.97,160.73,144.08,139.87,138.59, 131.30,128.94,128.68,128.11,119.88,118.90,115.04,113.78,98.49,55.33,50.97, 34.95,29.74,29.43,27.61.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₃₀N₅O₃:448.2349；found 448.2338.

IR(cm^-1):3469.03,3242.52,2959.32,2926.25,1618.90,1589.14,1562.60, 1512.59,1442.84,1429.01,1362.50,1325.73,1303.34,1255.14,1168.93,1068.02, 1031.51,1007.47,926.08,846.61,762.88,593.99.

Embodiment 4

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- is fluorine-based The synthesis of benzamide (I-4)

The synthesis of 4- (the fluoro- N- methylbenzoylamin. o of 4-) piperidines -1- carboxylate (VIII-4)

With 4- fluobenzoic acid (VII-4) (2.00g, 14.40mmol) for raw material, same VIII-1 is operated, chromatographs (petroleum through column Ether: ethyl acetate=6:1) separation, light yellow oil 3.68g is obtained, yield: 76%.

¹H-NMR(300MHz,CDCl₃)δ(ppm):7.53-7.34(m,2H,ArH),7.15-7.08(m,2H,ArH), 4.67(s,1H,CONCH),4.23(s,2H,CONCHCH₂ ),3.27-3.00(m,2H,CONCHCH₂ ),2.88(s,3H,NCH₃), 1.72(brs,4H,ONCHCH₂ ),1.49(s,9H,Boc-H).

The synthesis of fluoro- N- methyl-N- (piperidin-4-yl) benzamide (IX-4) of 4-

With VIII-4 (3.60g, 10.70mmol) for raw material, same IX-1 is operated, obtains light yellow oil 2.47g, yield: 98%.N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) fluorine-based benzene first of -4- The synthesis of amide (I-4)

With IV (500.00mg, 2.30mmol), IX-4 (543.00mg, 2.30mmol) is raw material, same I-1 is operated, through column (methylene chloride: methanol=200:1~100:1) separation is chromatographed, obtains gray solid, yield: 58%, m.p.87~88 DEG C.

¹H-NMR(300MHz,CDCl₃)δ(ppm):7.47-7.42(m,3H,ArH),7.28-7.20(m,3H,ArH,NH), 7.15-7.09(m,2H,ArH),6.89-6.80(m,2H,ArH),6.04(s,1H,ArH),5.02-4.60(m,1H, CH₃NCH),4.60-4.20(m,2H,CON(CH₂)₂ ),3.72(s,3H,CH₃),3.23-2.97(m,2H,CON(CH₂)₂ ),2.89 (s,3H,CH₃),1.92-1.80(m,4H,CH(CH₂)₂ ).

¹³C-NMR(75MHz,CDCl₃) δ (ppm): 170.75,170.01,163.34 (d, J=250.20Hz), 144.12, 139.86,138.57,132.59,131.35,129.00,128.11,119.73,118.91 115.63 (d, J=21.70Hz), 115.09,98.46,45.05,34.94,29.66,29.36,28.83.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₇FN₅O₂:436.2149；found 436.2141.

IR(cm^-1):3522.58,3271.66,2939.39,2859.62,1624.01,1557.39,1445.79, 1368.77,1325.66,1274.19,1222.40,1158.03,1065.48,1005.75,926.05,894.43,848.71, 760.00,729.00,583.85.

Embodiment 5

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- cyano The synthesis of benzamide (I-5)

The synthesis of 4- (4- cyano-N-methyl benzamido) piperidines -1- carboxylate (VIII-5)

With 4- cyanobenzoic acid (VII-5) (2.10g, 14.4mmol) for raw material, same VIII-1 is operated, chromatographs (stone through column Oily ether: ethyl acetate=6:1) separation, white solid 3.50g is obtained, yield: 71%, m.p.120~121 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 7.76 (d, J=8.00Hz, 2H, ArH), 7.51 (d, J=7.30Hz, 2H, ArH), 4.71 (s, 1H, CONCH),4.27(brs,2H,CONCHCH₂ ),3.00-2.89(m,1H,CONCHCH₂ ),2.81 (s,3H,NCH₃),2.51(s,1H,CONCHCH₂ ),1.77-1.71(m,4H,BocN(CH₂)₂ ),1.50(s,9H,Boc-H).

The synthesis of 4- cyano-N-methyl-N- (piperidin-4-yl) benzamide (IX-5)

With VIII-5 (3.67g, 10.70mmol) for raw material, same IX-1 is operated, obtains colorless oil 2.40g, yield: 92%.N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- cyano benzene first The synthesis of amide (I-5)

With IV (500.00mg, 2.30mmol), IX-5 (559.00mg, 2.30mmol) is raw material, same I-1 is operated, through column (methylene chloride: methanol=200:1~100:1) separation is chromatographed, obtains greenish yellow solid 559.00mg, yield: 55%, m.p.204 ~205 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 7.74 (d, J=7.50Hz, 2H, ArH), 7.61-7.35 (m, 3H, ArH),7.29-7.20(m,3H,ArH,NH),6.9-6.81(m,2H,ArH),6.04(s,1H,ArH),4.92-4.72(m,1H, CH₃NCH),4.69-4.24(m,2H,CON(CH₂)₂ ),3.72(s,3H,CH₃),3.22-2.97(m,2H,CON(CH₂)₂ ),2.83 (s,3H,CH₃),1.98-1.81(m,4H,CH(CH₂)₂ ).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):170.04,169.58,144.17,140.84,139.84,138.58, 132.49,131.42,128.11,127.57,127.48,118.91,117.94,115.13,113.58,98.43,51.55, 34.94,32.14,29.04,27.93.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₇N₆O₂:443.2195；found 443.2185.

IR(cm^-1):3294.45,2938.25,2868.83,2230.93,1504.66,1586.66,1556.58, 1447.14,1368.27,1329.95,1203.61,1069.66,1021.24,856.78,757.43,656.91,558.69.

Embodiment 6

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- trifluoro The synthesis of methyl benzamide (I-6)

The synthesis of 4- (N- methyl -4- (trifluoromethyl) benzamido) piperidines -1- carboxylate (VIII-6)

With 4- trifluoromethylbenzoic acid (VII-6) (2.70g, 14.40mmol) for raw material, same VIII-1 is operated, through column layer (petroleum ether: ethyl acetate=6:1) separation is analysed, obtains white solid 3.89g, yield: 70%, m.p.122~123 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 7.71 (d, J=8.00Hz, 2H, ArH), 7.52 (d, J=7.60Hz, 2H,ArH),4.73(s,1H,CONCH),4.27(s,2H,CONCHCH₂ ),3.00(s,1H,CONCHCH₂ ),2.81(s,3H, NCH₃),2.52(s,1H,CONCHCH₂ ),1.77(brs,4H,BocN(CH₂)₂ ),1.49(s,9H,Boc-H).

The synthesis of 4- trifluoromethyl-N- methyl-N- (piperidin-4-yl) benzamide (IX-6)

With VIII-6 (4.10g, 10.70mmol) for raw material, same IX-1 is operated, obtains colorless oil 2.88g, yield: 94%.N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- trifluoromethyl The synthesis of benzamide (I-6)

With IV (500.00mg, 2.30mmol), IX-6 (658.00mg, 2.30mmol) is raw material, same I-1 is operated, through column (methylene chloride: methanol=200:1~100:1) separation is chromatographed, obtains greenish yellow solid 703.00mg, yield: 63%, m.p.187 ~188 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 7.70 (d, J=7.40Hz, 2H, ArH), 7.60-7.38 (m, 3H, ArH),7.35-7.20(m,3H,ArH,NH),6.89-6.80(m,2H,ArH),6.04(s,1H,ArH),5.04-4.74(m, 1H,CH₃NCH),4.72-4.26(m,2H,CON(CH₂)₂ ),3.72(s,3H,CH₃),3.30-3.02(m,2H,CON(CH₂)₂ ), 2.85(s,3H,CH₃),2.06-1.82(m,4H,CH(CH₂)₂ ).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):172.68,170.02,144.14,140.08,139.85,138.57, (131.39,128.10,127.19,126.98,125.69,121.35 q, J=271.20Hz), 119.55,118.90, 115.10,98.45,51.34,34.93,32.05,30.03,29.14.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₇F₃N₅O₂:486.2117；found 486.2101.

IR(cm^-1):3326.03,2940.53,1624.30,1582.28,1556.33,1446.57,1369.62, 1325.64,1276.02,1170.31,1123.75,1071.10,1016.11,925.88,854.57,757.17,663.81.

Embodiment 7

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- acetyl The synthesis of yl-benzamide (I-7)

The synthesis of 4- (4- acetyl group-N- methylbenzoylamin. o) piperidines -1- carboxylate (VIII-7)

With 4- formylbenzoate (VII-7) (2.40g, 14.40mmol) for raw material, same VIII-1 is operated, is chromatographed through column (petroleum ether: ethyl acetate=6:1) separation, obtains faint yellow solid 2.70g, yield: 52%, m.p.101~102 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 8.02 (d, J=8.10Hz, 2H, ArH), 7.48 (d, J=7.60Hz, 2H,ArH),4.72(s,1H,CONCH),4.25(brs,2H,CONCHCH₂ ),2.94-2.89(m,1H,CONCHCH₂ ),2.80 (s,3H,NCH₃),2.64(s,3H,COCH₃),2.50-2.43(m,1H,CONCHCH₂ ),1.76(brs,4H,BocN(CH₂)₂ ), 1.48(s,9H,Boc-H).

The synthesis of 4- acetyl group-N- methyl-N- (piperidin-4-yl)-benzamide (IX-7)

With VIII-7 (3.85g, 10.70mmol) for raw material, same IX-1 is operated, obtains light yellow oil 2.58g, yield: 93%.N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- acetylbenzene The synthesis of formamide (I-7)

With IV (500.00mg, 2.30mmol), IX-7 (598.00mg, 2.30mmol) is raw material, same I-1 is operated, through column (methylene chloride: methanol=200:1~100:1) separation is chromatographed, obtains brown solid 538.00mg, yield: 51%, m.p.177~ 178℃。

¹H-NMR(300MHz,CDCl₃) δ (ppm): 8.02 (d, J=8.00Hz, 2H, ArH), 7.50-7.40 (m, 3H, ArH),7.40-7.13(m,3H,ArH,NH),6.89-6.80(m,2H,ArH),6.04(s,1H,ArH),5.03-4.72(m, 1H,CH₃NCH),4.69-4.28(m,2H,CON(CH₂)₂ ),3.72(s,3H,CH₃),3.30-3.02(m,2H,CON(CH₂)₂ ), 2.84(s,3H,CH₃),2.64(s,3H,COCH₃),2.03-1.70(m,4H,CH(CH₂)₂ ).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):172.63,170.56,170.01,144.15,140.92,139.85, 138.58,137.82,131.38,128.58,128.10,127.00,126.77,118.90,115.10,98.48,51.36, 34.94,32.04,29.67,29.19,26.62.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₆H₃₀N₅O₃:460.2349；found 460.2338.

IR(cm^-1):3327.23,2944.82,2865.06,1683.88,1616.87,1556.60,1504.68, 1468.81,1366.06,1327.17,1265.23,1079.76,1065.61,1013.66,863.06,851.80,757.45, 602.99.

Embodiment 8

N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- methyl sulfone The synthesis of yl-benzamide (I-8)

The synthesis of 4- (N- methyl -4- (mesyl) benzamido) piperidines -1- carboxylate (VIII-8)

With 4- thiamphenicol benzoic acid (VII-8) (3.10g, 14.40mmol) for raw material, same VIII-1 is operated, is chromatographed through column (petroleum ether: ethyl acetate=3:1) separation, obtains white solid 3.90g, yield: 69%, m.p.196~197 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 8.01 (d, J=8.10Hz, 2H, ArH), 7.57 (d, J=7.40Hz, 2H,ArH),4.69(m,1H,CONCH),4.44-4.00(m,2H,CONCHCH₂ ),3.07(s,3H,SO₂CH₃ ),3.03-2.81 (m,2H,CONCHCH₂ ),2.77(s,3H,NCH₃),1.85-1.60(m,4H,BocN(CH₂)₂ ),1.46(s,9H,Boc-H).

The synthesis of 4- methylsulfonyl-N- methyl-N- (piperidin-4-yl) benzamide (IX-8)

With VIII-8 (4.20g, 10.70mmol) for raw material, same IX-1 is operated, obtains colorless oil 3.00g, yield: 96%.N- methyl-N- (1- (2- ((1- methyl-1 H- pyrazoles -5- base) amino) benzoyl) piperidin-4-yl) -4- methylsulfonyl benzene The synthesis of formamide (I-8)

With IV (500.00mg, 2.30mmol), IX-8 (681.00mg, 2.30mmol) is raw material, same I-1 is operated, through column (methylene chloride: methanol=150:1) separation is chromatographed, obtains dark green solid 831.00mg, yield: 73%, m.p.107~108 DEG C.

¹H-NMR(300MHz,CDCl₃) δ (ppm): 8.03 (d, J=7.70Hz, 2H, ArH), 7.59 (d, J=7.80Hz, 2H,ArH),7.47(s,1H,NH),7.32-7.23(m,3H,ArH),6.89-6.80(m,2H,ArH),6.04(s,1H,ArH), 4.99-4.81(m,1H,CH₃NCH),4.68-4.23(m,2H,CON(CH₂)₂ ),3.72(s,3H,SO₂CH₃),3.23-2.95(m, 5H,CONCH₃,CON(CH₂)₂ ),2.83(s,3H,CH₃),2.06-1.62(m,4H,CH(CH₂)₂ ).

¹³C-NMR(125MHz,CDCl₃)δ(ppm):170.03,169.66,144.11,141.92,141.59,139.90, 138.53,131.43,128.13,128.01,127.88,127.76,118.96,115.16,98.40,51.53,44.39, 34.94,32.21,29.67,29.02.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₃₀N₅O₄S:496.2019；found 496.2004.

IR(cm^-1):3457.23,3343.06,2927.76,1624.62,1586.54,1556.50,1446.95, 1428.07,1369.76,1326.92,1312.89,1277.30,1152.58,1065.37,1006.97,957.64, 853.66,785.69,752.34,682.56,553.70.

Embodiment 9

N- methyl-N- (1- (2- (phenylamino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- fluorobenzamide (I- 9) synthesis of the fluoro- N- of synthesis 4- (1- (2- iodophenyl) piperidin-4-yl)-N- methyl -2- (trifluoromethyl) benzamide (XI)

Compound X (2.00g, 8.00mmol) is dissolved in methylene chloride (10.00ml), is cooled to 0 DEG C after 2 drop DMF are added, It instills oxalyl chloride (0.80ml, 9.60mmol), 30min is stirred at room temperature in dislocation, and being spin-dried for methylene chloride, to obtain acyl chlorides spare.

IX-1 (2.40g, 8.00mmol) is dissolved in methylene chloride (20ml), addition triethylamine (1.70ml, 12.00mmol), it is cooled to 0 DEG C, 5h is stirred at room temperature in dislocation after instilling the dichloromethane solution of acyl chlorides.TLC (petroleum ether: acetic acid second Ester=3:1) it detects and has reacted.1NHCl, saturated sodium bicarbonate aqueous solution, saturated salt solution washing reaction liquid, anhydrous sulphur are used respectively Sour sodium dries, filters, and is spin-dried for solvent, through column chromatography (petroleum ether: ethyl acetate=8:1) separation, obtains faint yellow solid 3.70g, Yield 86%, m.p.85~86 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.91-7.81(m,1H,ArH),7.45-7.36 (m,2H,ArH),7.36-7.28(m,2H,ArH),7.24-7.17(m,1H,ArH),7.14-7.02(m,1H,ArH),5.00- 4.75(m,1H,CH₃NCH),4.23(brs,2H,NCHCH₂ ),3.07(br,2H,NCHCH₂ ),2.58(s,3H,CH₃),1.75 (m,4H,CON(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.91-7.81(m,1H,ArH),7.45-7.36 (m,2H,ArH),7.36-7.28(m,2H,ArH),7.24-7.17(m,1H,ArH),7.14-7.02(m,1H,ArH),4.23 (brs,2H,NCHCH₂ ),3.38(m,1H,CH₃NCH),3.02(s,3H,CH₃),2.41(brs,2H,NCHCH₂ ),1.75(m, 4H,CON(CH₂)₂ ).

N- methyl-N- (1- (2- (phenylamino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- fluorobenzamide (I- 9) synthesis

By XI (500.00mg, 0.90mmol), aniline (XII-1) (84.00mg, 0.90mmol), palladium acetate (20.00mg, It 0.09mmol) is dissolved in toluene (10.00ml) with BINAP (112.00mg, 0.18mmol), 100 DEG C of stirrings is heated under nitrogen protection 12h.TLC (methylene chloride: methanol=35:1) detects raw material fully reacting.Reaction solution is cooling, it is spin-dried for after suction filtered through kieselguhr molten Agent obtains faint yellow solid 300.00mg through column chromatography (eluant, eluent: methylene chloride: methanol=300:1~100:1) separation, receives Rate: 67%, m.p.164~165 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.45-7.38(m,2H,ArH),7.30-7.20 (m,6H,ArH),7.14-7.03(m,2H,ArH),6.96-6.87(m,2H,ArH),4.88-4.78(m,1H,CH₃NCH), 4.67-4.00(m,2H,CON(CH₂)₂ ),3.16-2.91(m,2H,CON(CH₂)₂ ),2.47(s,3H,CH₃),1.76-1.58(m, 4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.45-7.38(m,2H,ArH),7.30-7.20 (m,6H,ArH),7.14-7.03(m,2H,ArH),6.96-6.87(m,2H,ArH),4.67-4.00(brs,2H,CON (CH₂)₂ ),3.4.-3.30(m,1H,CH₃NCH),2.83(s,3H,CH₃),2.72-2.56(m,2H,CON(CH₂)₂ ),1.76- 1.58(m,4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 169.35,167.44,161.60 (d, J= 250.10Hz),142.01,141.26,141.22,129.94,128.95,128.88,128.79,128.47,128.36, 127.31,123.64,120.78,119.68,118.97 (d, J=21.30Hz), 117.66,113.89 (dq, J=9.40, 4.20Hz),50.35,30.78,28.45,27.69.

Minor rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 169.33,167.44,161.60 (d, J= 250.10Hz),142.01,141.26,141.22,130.03,128.95,128.88,128.79,128.47,128.36, 127.15,123.64,120.97,119.68,118.97 (d, J=21.30Hz), 117.66,113.89 (dq, J=9.40, 4.20Hz),56.08,29.18,28.75,26.81.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₇H₂₆F₄N₃O₂:500.1961；found 500.1949.

IR(cm^-1):3044.25,2943.95,2867.26,1635.00,1594.22,1511.76,1450.29, 1325.89,1279.31,1173.89,1136.30,1040.50,995.21,906.81,877.35,877.35,691.71, 500.18.

Embodiment 10

N- methyl-N- (1- (2- ((4- cyano) phenylamino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- fluorobenzene The synthesis of formamide (I-10)

With XI (500.00mg, 0.90mmol), 4- cyano-aniline (XII-2) (106.00mg, 0.90mmol) is raw material, Same I-9 is operated, through column chromatography (eluant, eluent: methylene chloride: methanol=300:1~100:1) separation, obtains white powder 297.00mg, yield 63%, m.p.226~227 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.53(s,1H,ArH),7.48-7.45(m, 3H, ArH), 7.41-7.34 (m, 2H, ArH), 7.29-7.27 (m, 2H, ArH), 7.09 (t, J=6.00Hz, 1H, ArH), 7.04-7.01(m,2H,ArH),4.84-4.75(m,1H,CH₃NCH),4.68-3.70(m,2H,CON(CH₂)₂ ),3.24-2.90 (m,2H,CON(CH₂)₂ ),2.46(s,3H,CH₃),1.76-1.69(m,2H,NCH(CH₂)₂ ),1.69-1.58(m,2H,NCH (CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.53(s,1H,ArH),7.48-7.45(m, 3H, ArH), 7.41-7.34 (m, 2H, ArH), 7.29-7.27 (m, 2H, ArH), 7.09 (t, J=6.00Hz, 1H, ArH), 7.04-7.01(m,2H,ArH),4.68-3.70(m,2H,CON(CH₂)₂ ),3.42-3.27(m,1H,CH₃NCH),2.85(s, 3H.CH₃),2.72-2.57(m,2H,CON(CH₂)₂ ),1.76-1.69(m,2H,NCH(CH₂ )₂),1.69-1.58(m,2H,NCH (CH₂)₂ ).

Major rotamer:¹³C-NMR(125MHz,CDCl₃) δ (ppm): 169.05,167.96,162.14 (d, J= 251.40Hz), 147.27,138.81,133.73,131.63,131.31,130.56,129.31 (d, J=8.00Hz), 128.50 (q, J=7.50Hz), 128.04,124.9 (q, J=281.20Hz), 122.8,121.05,119.52 (d, J= 21.20Hz), 119.51,115.67,114.43 (d, J=22.50Hz), 102.13,50.82,31.27,28.88,28.23.

Minor rotamer:¹³C-NMR(125MHz,CDCl₃) δ (ppm): 169.05,167.53,162.21 (d, J= 250.00Hz), 146.95,139.02,133.73,131.63,131.31,130.70,129.31 (d, J=8.00Hz), 128.50 (q, J=7.50Hz), 127.96,124.9 (q, J=281.20Hz), 122.63,120.64,119.52 (d, J= 21.20Hz), 119.51,115.86,114.43 (d, J=22.50Hz), 102.45,56.42,28.88,28.23,27.26.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₈H₂₅F₄N₄O₂:525.1914found 525.1898.

IR(cm^-1):3492.63,3290.46,2955.75,2926.25,2215.65,1633.13,1583.44, 1516.89,1449.56,1411.71,1367.00,1319.46,1280.89,1161.66,1140.82,1040.35, 1009.21,904.82,873.92,846.23,740.45,544.11.

Embodiment 11

N- methyl-N- (1- (2- ((2- cyano-phenyl) amino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- fluorine The synthesis of benzamide (I-11)

With XI (500.00mg, 0.90mmol), 2- cyano-aniline (XII-3) (106.00mg, 0.90mmol) is raw material, behaviour Make same I-9, through column chromatography (eluant, eluent: methylene chloride: methanol=200:1~100:1) separation, obtain white powder 240.00mg, Yield 51%, m.p.187~188 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.54-7.49(m,1H,ArH),7.45-7.35 (m,4H,ArH),7.33-7.27(m,4H,ArH),7.13-7.08(m,1H,ArH),6.97-6.89(m,1H,ArH),4.88- 4.78(m,1H,CH₃NCH),4.60-3.46(m,2H,CON(CH₂)₂ ),3.25-2.95(m,2H,CON(CH₂)₂ ),2.55(s, 3H,CH₃),1.90-1.57(m,4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.54-7.49(m,1H,ArH),7.45-7.35 (m,4H,ArH),7.33-7.27(m,4H,ArH),7.13-7.08(m,1H,ArH),6.97-6.89(m,1H,ArH),4.60- 3.46(m,2H,CON(CH₂)₂ ),3.40-3.28(m,1H,CH₃NCH),2.89(s,3H,CH₃),2.70-2.64(m,2H,CON (CH₂)₂ ),1.90-1.57(m,4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 168.25,167.45,161.59 (d, J= 251.10Hz),145.71,138.20,133.40,132.75,131.29,129.96,128.94,128.83,128.48, (128.38,127.70,125.32 q, J=183.70Hz), 122.50,120.31,120.04,119.84,119.00 (d, J= 21.50Hz), 116.63,113.87 (dq, J=14.70,5.00Hz), 50.35,30.93,28.67,27.76.

Minor rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 168.25,167.45,161.59 (d, J= 251.10Hz),145.71,138.20,133.47,132.83,131.29,130.06,128.94,128.83,128.48, (128.38,127.50,125.32 q, J=183.70Hz), 122.42,120.31,120.04,119.84,119.00 (d, J= 21.50Hz), 116.63,113.87 (dq, J=14.70,5.00Hz), 56.04,28.67,27.76,26.86.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₈H₂₅F₄N₄O₂:525.1914；found 525.1899.

IR(cm^-1):3461.31,3300.38,2935.60,2219.24,1633.64,1589.08,1516.10, 1448.85,1422.55,1366.81,1326.76,1166.45,1122.68,1040.00,1005.65,905.71, 876.54,757.01.

Embodiment 12

N- methyl-N- (1- (2- ((4- methoxyphenyl) amino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- The synthesis of fluorobenzamide (I-12)

With XI (500.00mg, 0.90mmol), 4- aminoanisole (XII-4) (111.00mg, 0.90mmol) is raw material, Same I-9 is operated, through column chromatography (eluant, eluent: methylene chloride: methanol=200:1~100:1) separation, obtains white powder 476.00mg, yield 61%, m.p.165~166 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.45-7.39(m,1H,ArH),7.31(d,J =6.50Hz, 2H, ArH), 7.25-7.00 (m, 5H, ArH), 6.85 (d, J=8.90Hz, 2H, ArH), 6.82-6.74 (m, 1H, ArH),4.91-4.80(m,1H,CH₃NCH),4.72-4.20(m,2H,CON(CH₂)₂ ),3.78(s,3H,OCH₃),3.20- 2.96(m,2H,CON(CH₂)₂ ),2.57(s,3H,CH₃),1.81-1.59(m,4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.45-7.39(m,1H,ArH),7.31(d,J =6.50Hz, 2H, ArH), 7.25-7.00 (m, 5H, ArH), 6.85 (d, J=8.90Hz, 2H, ArH), 6.82-6.74 (m, 1H, ArH),4.72-4.20(m,2H,CON(CH₂)₂ ),3.78(s,3H,OCH₃),3.41-3.30(m,1H,CH₃NCH),2.92(s, 3H,CH₃),2.80-2.62(m,2H,CON(CH₂)₂ ),1.81-1.59(m,4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(125MHz,CDCl₃) δ (ppm): 170.18,167.94,162.12 (d, J= 250.00Hz),155.47,144.10,135.03,131.83,130.60,129.40,129.34,127.89,122.76(q,J =270.00Hz), 122.48,121.74,119.46 (d, J=21.60Hz), 118.51,115.73,114.74,114.40 (d, ), J=24.80Hz 55.56,51.00,31.42,29.05,28.39.

Minor rotamer:¹³C-NMR(125MHz,CDCl₃) δ (ppm): 170.25,167.58,162.18 (d, J= 249.00Hz),155.57,144.13,134.88,131.91,130.69,129.40,129.34,127.73,122.76(q,J =270.00Hz), 122.48,121.50,119.46 (d, J=21.60Hz), 118.51 (s), 115.73,114.74,114.40 (d, J=24.80Hz), 56.66,55.56,29.84,29.39,27.39.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₈H₂₈F₄N₃O₃:530.2067；found 530.2056.

IR(cm^-1):3374.40,2934.04,1636.33,1587.02,1513.36,1367.21,1328.60, 1318.48,1247.04,1170.35,1136.40,1038.64,1004.04,905.98,876.43,767.36,749.28, 519.68.

Embodiment 13

N- methyl-N- (1- (2- ((2- methoxyphenyl) amino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- The synthesis of fluorobenzamide (I-13)

With XI (500.00mg, 0.90mmol), 2- aminoanisole (XII-5) (111.00mg, 0.90mmol) is raw material, Same I-9 is operated, through column chromatography (eluant, eluent: methylene chloride: methanol=300:1~100:1) separation, obtains white powder 276.00mg, yield 58%, m.p.219~220 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.45-7.35(m,2H,ArH),7.29-7.25 (m,4H,ArH),7.17-7.00(m,1H,ArH),7.00-6.91(m,1H,ArH),6.88-6.84(m,3H,ArH),5.10- 4.10(m,3H,CH₃NCH,CON(CH₂)₂ ),3.86(s,3H,OCH₃),3.16-2.90(m,2H,CON(CH₂)₂ ),2.44(s, 3H,CH₃),1.85-1.47(m,4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):7.45-7.35(m,2H,ArH),7.29-7.25 (m,4H,ArH),7.17-7.00(m,1H,ArH),7.00-6.91(m,1H,ArH),6.88-6.84(m,3H,ArH),5.08- 4.09(m,2H,CON(CH₂)₂ ),3.86(s,3H,OCH₃),3.38-3.23(m,1H,CH₃NCH),2.77(s,3H,CH₃), 2.69-2.61(m,2H,CON(CH₂)₂ ),1.85-1.47(m,4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 169.07,167.40,161.58 (d, J= 250.80Hz),148.41,140.33,131.75,129.70,128.91,128.80,128.35,127.28,127.16, (120.24,120.08,120.10,120.04,118.98 d, J=21.10Hz), 118.09,114.90,113.87 (ddd, J= 13.80,8.80,4.10Hz),110.24,55.21,50.34,30.66,28.36,27.59.

Minor rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 169.07,167.40,161.58 (d, J= 250.80Hz),148.41,140.33,131.75,129.77,128.91,128.80,128.35,127.16,127.12, (120.24,120.08,120.10,120.04,118.98 d, J=21.10Hz), 118.09,114.90,113.87 (ddd, J= 13.80,8.80,4.10Hz),110.32,56.08,55.21,29.09,28.63,26.65.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₈H₂₈F₄N₃O₃:530.2067；found 530.2058.

IR(cm^-1):3355.27,2991.15,1639.17,1589.19,1516.19,1420.47,1367.24, 1327.27,1278.32,1244.99,1169.44,1130.41,1074.06,1039.91,906.12,874.10,751.72, 642.40,606.02,560.94.

Embodiment 14

N- methyl-N- (1- (2- (pyridin-4-yl amino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- fluorobenzene first The synthesis of amide (I-14) is with XI (500.00mg, 0.90mmol), 4-aminopyridine (XII-6) (85.00mg, 0.90mmol) Raw material operates same I-9, through column chromatography (eluant, eluent: methylene chloride: methanol=150:1~100:1) separation, obtains pale yellow powder 198.00mg, yield 44%, m.p.136~137 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):8.40-8.19(m,2H,ArH),7.49-7.29 (m,7H,ArH,NH),7.21-7.15(m,1H,ArH),6.93-6.78(m,2H,ArH),5.07-4.48(m,2H,CH₃NCH, CON(CH₂)₂ ),4.19-3.68(m,1H,CON(CH₂)₂ ),3.25-2.91(m,2H,CON(CH₂)₂),2.41(s,3H,CH₃), 1.92-1.43(m,4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):8.40-8.19(m,2H,ArH),7.49-7.29 (m,7H,ArH,NH),7.21-7.15(m,1H,ArH),6.93-6.78(m,2H,ArH),5.07-4.48(m,1H,CON (CH₂)₂ ),4.19-3.68(m,1H,CON(CH₂)₂ ),3.42-3.27(m,1H,CH₃NCH),2.85(s,3H,CH₃),2.69- 2.58(m,2H,CON(CH₂)₂ ),1.92-1.43(m,4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 168.88,167.95,162.13 (d, J= 247.60Hz), 150.24,131.63,130.42,129.35,129.24,128.89 (d, J=7.90Hz), 127.85, (123.53,123.32,122.33,122.00,119.45 d, J=21.20Hz), 114.38 (d, J=20.20Hz), 109.94, 109.80,50.70,31.17,28.81,28.15.

Minor rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 168.88,167.95,162.13 (d, J= 247.60Hz), 150.24,131.63,130.57,129.35,129.24,128.89 (d, J=7.90Hz), 127.85, (123.53,123.32,122.33,122.00,119.45 d, J=21.20Hz), 114.38 (d, J=20.20Hz), 109.94, 109.80,56.39,29.58,29.19,27.24.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₆H₂₅F₄N₄O₂:501.1914；found 501.1900.

IR(cm^-1):3263.74,2930.64,1631.34,1592.68,1515.28,1422.00,1410.90, 1370.34,1321.60,1282.23,1267.24,1214.83,1171.03,1134.24,1072.83,1041.27, 997.24,905.95,817.03,757.23,528.80.

Embodiment 15

N- methyl-N- (1- (2- (pyridine -2- base amino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- fluorobenzene first The synthesis of amide (I-15) is with XI (500.00mg, 0.90mmol), 2-aminopyridine (XII-7) (85.00mg, 0.90mmol) Raw material operates same I-9, through column chromatography (eluant, eluent: methylene chloride: methanol=150:1~100:1) separation, obtains pale yellow powder 162.00mg yield 36%, m.p.155~156 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):8.30-8.16(m,1H,ArH),8.01(d,J =8.3Hz, 1H, ArH), 7.84-7.66 (m, 2H, ArH), 7.57-7.25 (m, 5H, ArH, NH), 7.17-6.96 (m, 1H, ArH),6.96-6.67(m,2H,ArH),4.89-4.77(m,1H,CH₃NCH),4.74-3.65(m,2H,CON(CH₂)₂ ), 3.21-2.93(m,2H,CON(CH₂)₂ ),2.50(s,3H,CH₃),1.93-1.50(m,4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):8.30-8.16(m,1H,ArH),8.01(d,J =8.30Hz, 1H, ArH), 7.84-7.66 (m, 2H, ArH, NH), 7.57-7.25 (m, 5H, ArH), 7.17-6.96 (m, 1H, ArH),6.96-6.67(m,2H,ArH),4.74-3.65(m,2H,CON(CH₂)₂ ),3.42-3.23(m,1H,CH₃NCH),2.84 (s,3H,CH₃),2.69-2.61(m,2H,CON(CH₂)₂ ),1.93-1.50(m,4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 122.64,169.54,162.13 (d, J= 258.70Hz),155.39,148.07,148.01,138.77,137.66,130.43,129.38,129.28,128.96, (128.86,127.50,121.82,121.35,119.42 d, J=20.90Hz), 115.37,114.50 (d, J=4.50Hz), 109.98,50.84,31.26,28.94,28.16.

Minor rotamer:¹³C-NMR(75MHz,CDCl₃) δ (ppm): 122.64,169.54,162.13 (d, J= 258.70Hz),155.39,148.07,148.01,138.77,137.75,130.52,129.38,129.28,128.96, (128.86,127.37,121.82,121.35,119.42 d, J=20.90Hz), 115.54,114.17 (d, J=4.10Hz), 109.98,56.50,29.16,28.45,27.23.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₆H₂₅F₄N₄O₂:501.1914；found 501.1903.

IR(cm^-1):3392.33,2920.35,2861.36,2861.36,1592.16,1511.12,1434.99, 1367.46,1329.28,1307.24,1173.57,1132.21,1075.27,1038.87,907.50,862.63,759.68, 644.57,603.31,523.19.

Embodiment 16

N- methyl-N- (1- (2- ([1,1 '-diphenyl] -4- base amino) benzoyl) piperidin-4-yl) -2- fluoroform The synthesis of base -4- fluorobenzamide (I-16)

With XI (500.00mg, 0.90mmol), 4- phenylaniline (XII-8) (152.00mg, 0.90mmol) is raw material, behaviour Make same I-9, through column chromatography (eluant, eluent: methylene chloride: methanol=300:1~100:1) separation, obtain yellow powder 340.00mg, Yield 66%, m.p.112~113 DEG C.

Major rotamer:¹H-NMR(500MHz,CDCl₃)δ(ppm):7.62-7.49(m,5H,ArH),7.49-7.38 (m, 3H, ArH), 7.38-7.25 (m, 5H, ArH, NH), 7.19-7.16 (m, 3H, ArH), 6.99 (t, J=7.50Hz, 1H, ArH),4.91-4.84(m,1H,CH₃NCH),4.82-3.79(m,2H,CON(CH₂)₂ ),3.30-2.95(m,2H,CON (CH₂)₂ ),2.52(s,3H,CH₃),1.84-1.73(m,2H,NCH(CH₂)₂ ),1.69-1.62(m,2H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(500MHz,CDCl₃)δ(ppm):7.62-7.49(m,5H,ArH),7.49-7.38 (m, 3H, ArH), 7.38-7.25 (m, 5H, ArH, NH), 7.19-7.16 (m, 3H, ArH), 6.99 (t, J=7.50Hz, 1H, ArH),4.82-3.79(m,2H,CON(CH₂)₂ ),3.47-3.28(m,1H,CH₃NCH),2.89(s,3H,CH₃ ),2.73-2.64 (m,2H,CON(CH₂)₂ ),1.84-1.73(m,2H,NCH(CH₂)₂ ),1.69-1.62(m,2H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(125MHz,CDCl₃) δ (ppm): 169.81,167.92,162.09 (d, J= 249.87Hz), 141.99,141.62,140.69,134.08,131.77 (dd, J=4.30,2.30Hz), 130.48,129.32 (d, J=8.10Hz), 128.92 (dd, J=16.10,7.30Hz), 128.72,127.97,127.88,126.67,126.46, 122.99 (q, J=261.25Hz), 120.33,119.43 (d, J=20.80Hz), 118.49,118.28,118.24,114.35 (dd, J=24.80,4.60Hz), 50.89,31.27,28.94,28.21.

Minor rotamer:¹³C-NMR(125MHz,CDCl₃) δ (ppm): 169.77,167.54,162.21 (d, J= 250.25Hz), 141.81,141.66,140.63,134.31,131.77 (dd, J=4.30,2.30Hz), 130.58,129.32 (d, J=8.10Hz), 128.92 (dd, J=16.10,7.30Hz), 128.77,128.03,127.71,126.75,126.50, (122.99 q, J=261.250Hz), 120.33,119.43 (d, J=20.80Hz), 118.49,118.28,118.24, 114.35 (dd, J=24.80,4.60Hz), 56.59,29.72,29.31,27.27.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₃₃H₃₀F₄N₃O₂:576.2274；found 576.2257.

IR(cm^-1):3376.30,3028.90,2930.00,2858.81,1631.67,1596.56,1523.77, 1423.06,1369.58,1320.85,1279.21,1170.45,1134.23,1072.03,1040.87,1005.23, 905.16,838.02,762.81,698.13,501.94.

Embodiment 17

N- methyl-N- (1- (2- ((4- Fonnylphenyl) amino) benzoyl) piperidin-4-yl) -2- trifluoromethyl -4- The synthesis of fluorobenzamide (I-17)

With XI (500.00mg, 0.90mmol), 4-aminobenzoic acid (XII-9) (123.00mg, 0.90mmol) is raw material, Same I-9 is operated, through column chromatography (eluant, eluent: methylene chloride: methanol=150:1~100:1) separation, obtains white powder 210.00mg yield 43%, m.p.178~179 DEG C.

Major rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):8.16-7.92(m,2H,ArH),7.55-7.51 (m,2H,ArH),7.46-7.30(m,4H,ArH),7.22-7.01(m,3H,ArH),4.87-4.79(m,1H,CH₃NCH), 4.40-3.54(m,2H,CON(CH₂)₂ ),3.17-2.93(m,2H,CON(CH₂)₂ ),2.41(s,3H,CH₃),1.93-1.41(m, 4H,NCH(CH₂)₂ ).

Minor rotamer:¹H-NMR(300MHz,CDCl₃)δ(ppm):8.16-7.92(m,2H,ArH),7.55-7.51 (m,2H,ArH),7.46-7.30(m,4H,ArH),7.22-7.01(m,3H,ArH),4.40-3.54(m,2H,CON(CH₂)₂ ), 3.39-3.32(m,1H,CH₃NCH),3.17-2.93(m,2H,CON(CH₂)₂ ),2.80(s,3H,CH₃),1.93-1.41(m, 4H,NCH(CH₂)₂ ).

Major rotamer:¹³C-NMR(75MHz,CDCl₃)δ(ppm):170.84,169.34,167.99,162.09 (d, J=250.50Hz), 156.98,148.45,139.09,132.16,130.46,129.36,129.26,128.98, 127.81,124.42 (q, J=206.30Hz), 122.78,121.53,120.43,119.46 (d, J=20.80Hz), (114.88,114.37 d, J=20.60Hz), 50.73,31.16,28.81,28.10.

Minor rotamer:¹³C-NMR(75MHz,CDCl₃)δ(ppm):170.72,169.34,167.99,162.09 (d, J=250.50Hz), 156.98,148.45,139.09,132.16,130.58,129.36,129.26,128.98, 127.68,124.42 (q, J=206.30Hz), 122.78,121.32,120.74,119.46 (d, J=20.80Hz), (114.88,114.37 d, J=20.60Hz), 56.41,29.63,29.15,27.20.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₈H₂₆F₄N₃O₄:544.1859；found 544.1845.

IR(cm^-1):3474.93,3307.19,3067.85,2933.28,2507.37,1707.76,1522.02, 1593.82,1451.29,1411.14,1370.89,1326.58,1280.08,1173.88,1134.95,1072.94, 1041.42,905.55,844.43,769.54,750.47,660.97,500.78.

The pharmacological experiment and result of part of compounds obtained by above-mentioned preparation are as follows:

One, Gli-luciferase is tested

1. experimental principle

The firefly element enzyme gene for being driven one by 12xGLI-binding site with slow virus stable transfection mode Firefly is imported in NIH/3T3 cell, while the renilla luciferase gene renilla that a CMV promoter drives being led Enter.Wherein firefly is used to detect the transcriptional activity of Hedgehog signal, reference gene of the renilla as reporter gene.With Hedgehog ligand can significantly improve the Hedgehog access transcriptional activity of the cell line.

2. experimental material

ShhFlashII/NIH3T3 cell line: Life Technologies

96 orifice plates: Corning Costar#3595

DMEM culture medium: Life Technologies

Luciferase Substrate:Promega

Stop&Substrate:Promega

Microplate reader: Themo Electro Co.

CO2 incubator: ESCO Micro Pte.Ltd.

Cell counter: Themo Electro Co.

3. experimental method

(1) the ShhFlashII/NIH3T3 single cell suspension containing pancreatin will be bought and carries out cell count.

The every hole of (2) 96 orifice plates is inoculated with 100 μ l and contains 2x10⁴The cell of quantity.

(3) by cell at 37 DEG C, 95% humidity, 5%CO₂Incubator in cultivate 3 days.

(4) the DMEM culture solution containing determinand or shh ligand is configured.

(5) cell culture fluid is sucked, determinand culture solution culture 48h is added.

(6) culture solution is removed, 50 holes μ l/ are addedLuciferase Substrate, is darkling cultivated 10min。

(7) Gli-luciferase reading is read with microplate reader.

(8) 50 μ l Stop& are addedSubstrate darkling cultivates 10min.

(9) Renilla Luciferase reading is read with microplate reader.

(10) reading of the Firefly reading of internal standard Renilla is normalized, calculates IC₅₀。

4. experimental result

The screening that the inhibitory activity of Hedeghog access is carried out to part of compounds of the present invention, the results are shown in Table 1.

IC of the 1. part test-compound of table to Gli-luciferase₅₀Value

Table 1 is the results show that the compounds of this invention has preferable inhibitory activity to Hedeghog access, wherein compound I-9 It is active best, to the IC of Gli-luc reporter₅₀Value is 34.07nM.

Two, Doay cell line vitality test is tested

1. test medicine

Compound I-1, I-9, I-13, LY2940680

2. experimental material

Fetal calf serum (Fetal Bovine Serum, FBS) (GIBCO, Invitrogen Corporation, NY, USA), DMEM culture medium (GIBCO, Invitrogen Corporation, NY, USA), penicillin (Sigma, St.Louis, MO, USA), streptomysin (Sigma, St.Louis, MO, USA), MTT reagent, green skies biotechnology research institute.

FORMA700 type ultra low temperature freezer, Thermo company；YC-300L type drug storage cabinet, middle section's U.S. water chestnut low temperature science and technology Co., Ltd；Direct-Q with pump type ultrapure water instrument, Millopore company；The work of SW-CJ-2FD type ultrapurification Platform: Purifying Equipment Co., Ltd., Suzhou；3111 type water-jacket typ CO of Forma₂Incubator: Thermo Electron company； Berthold LB941 micro-pore plate type multi-function microplate reader, Berthold company.

Doay cell line purchased from American Tissue Culture Collection (ATCC, Rockville, MD,USA)。

3. experimental method

Test grouping situation and drug concentration selection

Compound I (compound I-1, I-9, I-13) concentration selection: 0 μM, 0.01 μM, 0.10 μM, 1 μM, 5 μM, 10 μM.

Compound uses DMSO hydrotropy, and 0 μM is, as blank control group, the final concentration of DMSO does not surpass containing isometric DMSO Cross 0.10% (v/v).

Cell culture condition

Cell secondary culture 10-15 generation, condition of culture are (dense eventually containing penicillin (final concentration of 100U/ml), streptomysin Degree is 100 μ g/ml), the DMEM culture medium of 10%FBS discards old culture medium, washed with 2ml PBS when cell fusion is to 90% It washs cell 2 times, discards 0.25% (w/v) Trypsin-0.53mM EDTA mixture slaking liquid of addition 2ml after PBS, set microscope Lower observation, about 30s are rapidly added 2ml complete medium after cell rounding and terminate digestion, gently blow and beat, collect cell. 800rpm, is centrifuged 5min, discards supernatant liquid by 4 DEG C, cell is resuspended with complete medium, liquid is changed in sub-bottle culture every other day.

MTT test

By logarithmic growth phase cell with 1 × 10⁵The hole cells/ is inoculated in 96 orifice plates, is placed in 37 DEG C, 5%CO₂Under the conditions of train It supports, until being incubated for 2h with the DMEM culture medium of serum-free makes cell synchronization after cell 90% merges.Then, it discards supernatant, point Not Jia Ru concentration be that the complete DMEM culture medium of HG-I class compound of (0 μM, 0.01 μM, 0.10 μM, 1 μM, 5 μM, 10 μM) is incubated for 72h.Incubation terminates preceding 4h, and 20 μ l MTT solution (5mg/ml) are added in every hole.After incubation, each hole supernatant, every hole are discarded 150 μ l DMSO are added, 10min is vibrated on cell oscillation instrument, object to be crystallized measures OD with microplate reader after completely dissolution₅₇₀。

Inhibiting rate=(control group OD value-experimental group OD value)/control group OD value × 100%.

4. experimental result

People's marrow mother oncocyte (Daoy) external antiproliferative activity test is carried out to part of compounds of the present invention, the results are shown in Table 2.

Antiproliferative effect of 2. test-compound of table to people's marrow mother's oncocyte Daoy

The results show that test-compound has good inhibiting effect to the proliferation of people's medulloblastoma cell Daoy, The antiproliferative effect of middle compound I-1, I-9 and I-13 are better than positive control LY2940680.

Claims (10)

1. a kind of benzoyl piperidine analog derivative or its pharmaceutically acceptable salt as shown in general formula I:

Wherein:

R¹Selected from substituted or non-substituted phenyl, wherein substituted phenyl is by one, two or more substitutions chosen from the followings The phenyl that base replaces: halogen ,-NO₂、-NH₂,-CN, C1-C6 alkoxy ,-CHO ,-CF₃、-CONH₂, C1-C6 alkanoyl ,- SO₂CH₃、-NHCOCH₃、-NHCOCH₂CH₃、-CH₂CONH₂Or-NHCONH₂；

R²Selected from substituted or non-substituted five yuan or hexa-atomic aromatic heterocyclic or substituted or non-substituted C6-C12 aryl, In, substituted five yuan or hexa-atomic aromatic heterocyclic are five yuan or six replaced by one, two or more substituent groups chosen from the followings First aromatic heterocyclic: halogen, C1-C6 alkyl ,-CN ,-NH₂、-NHCOCH₃, C1-C6 alkanoyl, C1-C6 alkoxy or-COOH；It takes The C6-C12 aryl in generation is the C6-C12 aryl replaced by one, two or more substituent groups chosen from the followings: halogen, C1-C6 alkyl ,-CN ,-NH₂、-NHCOCH₃, C1-C6 alkanoyl, C1-C6 alkoxy or-COOH.

2. benzoyl piperidine analog derivative according to claim 1, it is characterised in that:

R¹It is selected fromWherein R³And R⁴It is each independently selected from-H, halogen ,-NO₂、-NH₂、-CN、-OCH₃、-CHO、- CF₃、-CONH₂、-COCH₃、-SO₂CH₃、-NHCOCH₃、-NHCOCH₂CH₃、-CH₂CONH₂Or-NHCONH₂；

R²Selected from substituted or non-substituted five yuan or hexa-atomic aromatic heterocyclic containing N or substituted or non-substituted phenyl or xenyl, Wherein, substituted five yuan or hexa-atomic aromatic heterocyclic containing N are five yuan replaced by one, two or more substituent groups chosen from the followings Or hexa-atomic aromatic heterocyclic containing N: halogen ,-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、-CN、-NH₂、-NHCOCH₃、-COCH₃、- COCH₂CH₃、-COCH₂CH₂CH₃、-OCH₃、-OCH₂CH₃、-OCH₂CH₂CH₃Or-COOH；Substituted phenyl or xenyl is by one The phenyl or xenyl that a, two or more substituent groups chosen from the followings replace: halogen ,-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、- CN、-NH₂、-NHCOCH₃、-COCH₃、-COCH₂CH₃、-COCH₂CH₂CH₃、-OCH₃、-OCH₂CH₃、-OCH₂CH₂CH₃Or-COOH.

3. benzoyl piperidine analog derivative according to claim 1, it is characterised in that:

R¹It is selected fromWherein R³Selected from-H, halogen ,-NO₂、-NH₂、-CN、-OCH₃、-CHO、-CF₃、-CONH₂、- COCH₃、-SO₂CH₃、-NHCOCH₃、-NHCOCH₂CH₃、-CH₂CONH₂Or-NHCONH₂；R⁴Selected from-H or-CF₃；

R²Selected from substituted or non-substituted five yuan or hexa-atomic aromatic heterocyclic containing N or substituted or non-substituted phenyl or xenyl, Wherein, substituted five yuan or hexa-atomic aromatic heterocyclic containing N are five yuan replaced by one, two or more substituent groups chosen from the followings Or hexa-atomic aromatic heterocyclic containing N: halogen ,-CH₃、-CN、-NH₂、-NHCOCH₃、-COCH₃、-OCH₃Or-COOH；Substituted phenyl or Xenyl is the phenyl or xenyl replaced by one, two or more substituent groups chosen from the followings: halogen ,-CH₃、-CN、- NH₂、-NHCOCH₃、-COCH₃、-OCH₃Or-COOH.

4. benzoyl piperidine analog derivative according to claim 1, it is characterised in that:

R¹It is selected from

R²It is selected from

5. benzoyl piperidine analog derivative according to claim 1, which is characterized in that the benzoyl piperidine class is spread out Biology is selected from I-1 to I-17:

6. the preparation method of any one of the claim 1-5 benzoyl piperidine analog derivative, which is characterized in that including by changing Object IV and IX is closed through condensation reaction prepare compound I:

7. the preparation method of any one of the claim 1-5 benzoyl piperidine analog derivative, which is characterized in that including by changing It closes object XI and reacts prepare compound I through Buchwald-Hartwig with XII:

Wherein, A is halogen.

8. a kind of pharmaceutical composition, which is characterized in that it includes the described in any item benzoyl piperidine classes of claim 1-5 to spread out Biology or its pharmaceutically acceptable salt and pharmaceutically acceptable auxiliary material.

9. any one of the claim 1-5 benzoyl piperidine analog derivative or its pharmaceutically acceptable salt are in preparation SMO Purposes in protein inhibitor drug.

10. any one of the claim 1-5 benzoyl piperidine analog derivative or its pharmaceutically acceptable salt are anti-in preparation Purposes in tumour medicine.