CN104860869A

CN104860869A - Compound with MEK kinase inhibition function, and preparation method and application thereof

Info

Publication number: CN104860869A
Application number: CN201510158186.8A
Authority: CN
Inventors: 徐萍; 牛彦; 黄文杰; 许凤荣
Original assignee: Peking University
Current assignee: Peking University
Priority date: 2015-04-03
Filing date: 2015-04-03
Publication date: 2015-08-26
Anticipated expiration: 2035-04-03
Also published as: CN104860869B

Abstract

The invention discloses a compound with an MEK kinase inhibition function, and a preparation method and application thereof. The chemical structure of the compound is as shown in a formula I which is described in the specification. Moreover, the invention provides the preparation method for the compound. Research shows that the compound has good MEK kinase inhibition activity and has wide application values in fields related to an MEK kinases inhibitor.

Description

There is compound of MEK kinase inhibition function and preparation method thereof and application

Technical field

The present invention relates to compound and preparation method thereof and application, particularly relate to compound with MEK kinase inhibition function and preparation method thereof and application.Belong to medicinal chemistry arts.

Background technology

Mitogen activated protein kinase (mitogen-activated protein kinase, MAPK) is the protein kinase with vital role found 1980 mid-nineties 90s, is core signal path important in cell.MAPK path is mainly divided into four class subtribes: extracellular signal-regulated kinase ERK (extracellular signal-regulated kinase) path, JNK path, p38 path and ERK5 path.ERK path is an important family in MAPK path, primarily of three core protein kinases RAF, MEK (Mitogen-activated and Extracellularsignal-regulated Kinase) and ERK composition, mutual Sequential Activation, the signal transmission network of a common composition precise and high efficiency plays its regulating effect.In resting cell, ERK is silent oscillation, after it accepts the phosphorylation of upstream activator molecule, namely become activated state.ERK accepts the signal of membrane receptor conversion and transmission and brings in nucleus by signal, makes some transcription factors (as Elk-1, c-fos etc.) phosphorylation in core, eventually through the adjustment phosphorylation level of transcription factor and then the expression level of regulatory gene; In addition, ERK also can make the many substrate phosphorylations in tenuigenin, and then affects structure and other the signal transduction pathway of cytoskeleton.

RAF/MEK/ERK signal path mainly participates in the significant process such as cell development, growth, differentiation, and its function raises with the generation of tumour closely related.So suppress this path one of research focus becoming new type antineoplastic medicine already, as RAF inhibitor Xarelto ratified listing, for the treatment of the noumenal tumours such as advanced renal cell cancer in 2005 by FDA.Some RAF inhibitor and mek inhibitor is also had to be in antitumor clinical experiment at present.And more and more study confirmation, this path also plays extremely important effect at virus infection with in copying.This path of selective exclusion can stop the breeding of multiple virus, comprises influenza virus, hsv, hepatitis virus, vaccinia virus, immunodeficiency virus, vesicular stomatitis virus etc.In addition, inflammation and many other diseases are as also relevant with the rise of ERK path in the generation of neuropathic pain, chronic obstructive pulmonary disease, hypertrophic cardiomyopathy, the heart method Theo skin (CFC) syndrome, Huntington chorea etc., development.

MEK1 and MEK2 is two and is closely related, and has the tyrosine/Serineprotein kinase of dual specific, in RAF/MEK/ERK signal path, plays vital effect.MEK1 and MEK2 has the amino acid identity of 79%, and has the ability of equal phosphorylated CREB substrate.Known ERK1/2 is unique substrate of MEK1/2, and MEK1/2 is the enzyme that uniquely can activate ERK1/2, MEK1 and MEK2 is the important tie point of ERK path.This point is very special, imply that the effect of mek inhibitor blocking-up ERK path will be high degree of specificity.Therefore, very promising strategy in new drug development is become to the suppression of MEK1/2 activity.

Reportedly be mek inhibitor with the compound of MEK specific binding, two classes can be divided into by mechanism of action: ATP competitive inhibitor and ATP noncompetitive inhibitor.

The competitive mek inhibitor of ATP and ATP act on same binding site, and vie each other with ATP, action effect is easily subject to the impact of ATP concentration.Because this inhibitor molecules amount is comparatively large, not high to kinase whose selectivity, can not specific effect be produced, so toxic side effect may be made larger.The competitive mek inhibitor of ATP mainly comprises cyano quinolines class and natural product.

The mechanism of action of ATP noncompetitive kinase inhibitor at least can be divided into two kinds: a kind of inhibitor directly with substrate competition specific binding site, another kind produces restraining effect by allosterism.Allosteric inhibitor directly with substrate competition binding site, but the position that is combined in enzyme is different from avtive spot, makes conformation transition and is fixed on a nonactive conformational state, and then the combination of prevention protein and substrate.Existing ATP noncompetitive mek inhibitor all belongs to allosteric inhibitor, is characterized in: 1. not kinase whose with ATP competitive target ATP-binding site, does not also compete MEK binding site with ERK.2. after being combined with MEK, modify the three-dimensional structure changing MEK, make MEK by upstream kinases phosphorylation, or not suppress phosphate to transfer to the avtive spot of ERK.3. its combining site does not have homologous sequence on other kinases, is that in all kinase inhibitor, specificity is the highest.4. restraining effect is reversible, and after stopping using, kinases can be reactivated.This kind of inhibitor generally has good cytoactive, this may be due to they by the micromolar ATP concentration of cytosol impact caused by.

Summary of the invention

What the object of this invention is to provide a kind of novel structure has compound of MEK kinase inhibition function and preparation method thereof.

The compound with MEK1 inhibit activities provided by the invention, and pharmacy acceptable salt or prodrug, is characterized in that the structure of described compound is such as formula shown in I:

Wherein, A ring is phenyl ring or tetrahydrobenzene;

R is hydrogen atom, C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group, C ₂-C ₇alkenyl or alkynyl, C ₃-C ₇cycloalkyl, tertiary butyloxycarbonyl acyl group, benzyloxy carbonyl acyl group, benzyl, substituted benzyl, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl;

R ₁for hydrogen, halogen, hydroxyl, cyano group, C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group, C ₂-C ₇alkenyl or alkynyl, replace or unsubstituted cycloalkyl;

R ₂for the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl;

R ₃for hydrogen, halogen, hydroxyl, C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group, C ₂-C ₇alkenyl or alkynyl, replace or unsubstituted cycloalkyl, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl;

X is oxygen, sulphur, carbon, NH, SO, SO ₂;

N is 1,2,3 or 4;

R ₄for hydroxyl, cyano group, carboxyl ,-COOR ₇,-CONR ₆r ₇, 1,2,4-oxadiazole, 1,3,4-oxadiazole, tetrazole, amino-1,3, the 4-oxadiazole of 5-, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl;

R ₆, R ₇be selected from hydrogen separately respectively, C _1-3alkyl.

In the present invention, preferably, described C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group comprise methyl, ethyl, propyl group, sec.-propyl, normal-butyl, 2-methyl-propyl, 2-butyl, tertiary butyl, n-pentyl, methoxy methylol, cyclopropyl, Cvclopropvlmethvl, adamantyl and the monosubstituted or polysubstituted above-mentioned group through halogen, hydroxyl, amino, cyano group, nitro, carboxyl.

In the present invention, preferably, described C ₂-C ₇thiazolinyl be aliphatic alkyl containing the straight or branched containing 2-7 carbon atom in C=C key simultaneously chain, comprise vinyl, propenyl, butenyl, isobutenyl, 3-methyl-2-butene base, just pentenyl, heptenyl, cyclopropylethenyl, cyclohexyl propenyl and by single or multiple halogen, hydroxyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group; Described alkynyl refers to the aliphatic alkyl containing the straight or branched containing 2-7 carbon atom in carbon-to-carbon triple bond simultaneously chain, comprises ethynyl, proyl, positive butynyl, butynyl, 3-methyl-2-butyne base, just pentynyl etc. and through single or multiple halogen, hydroxyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

In the present invention, preferably, described substituted or unsubstituted aryl refers to 6-14 unit monocycle or bicyclic aromatic group, comprises phenyl, naphthyl and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, kharophen, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group, wherein C ₁-C ₄alkyl comprise methyl, ethyl.

In the present invention, preferably, described substituted or unsubstituted heteroaryl refers to the heteroatomic 4-10 unit's monocycle or the bicyclic aromatic group that are independently selected from N, O and S containing 1-5, comprises pyrryl, pyridyl, imidazolyl, indyl and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

In the present invention, preferably, described substituted or unsubstituted heterocyclic radical refers to that preferred, heteroatoms comprises N, O, S containing one or more heteroatomic compound in its ring structure.

In the present invention, preferably, described substituted or unsubstituted heterocyclic radical is saturated or the aromatics of fractional saturation, preferred, five yuan and single six-membered rings heterocyclic radical and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

In the present invention, preferably, described halogen comprises fluorine, chlorine, bromine and iodine.

In the present invention, preferably, described replacement or unsubstituted cycloalkyl refer to C ₃-C ₁₀saturated monocycle or many rings ring system, comprise cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl, adamantyl and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

Prepare above-described compound, and the method for pharmacy acceptable salt or prodrug, it is characterized in that in described formula I structure, A ring is phenyl ring, R ₃for the preparation method of the compound of hydroxyl, comprise the steps:

1) there is diazotization reaction generation diazonium salt in substituted aniline under Sodium Nitrite and concentrated hydrochloric acid condition, is reduced to the compound of formula II structure subsequently under tin protochloride condition;

2) there is Fischer indoles ring closure reaction at trifluoroacetic acid as the condition of solvent and hydroresorcinol in formula II structural compounds, obtains the compound of formula III structure;

3) there is chlorination in the compound of formula III structure under the condition of cupric chloride and lithium chloride, then under the condition of lithium chloride and Quilonum Retard, the compound that eliminative reaction obtains formula IV structure occurs;

4) there is the compound that chlorination obtains formula V structure in the compound of formula IV structure under the condition of N-chlorosuccinimide;

5) compound of formula V structure is oxidized to the compound of formula VI structure under the condition of Fremy ' s Salt;

6) compound of formula VI structure and sulphonamide use microwave heating to react the compound of production VII structure under the condition of titanium tetrachloride and triethylamine;

7) compound of formula VII structure and nucleophilic reagent generation substitution reaction reduce that to obtain A ring in formula I structure be phenyl ring subsequently with vat powder, R ₃for the compound of hydroxyl;

Wherein, R is hydrogen atom, C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group, the alkenyl or alkynyl of C2-C7, replaces or unsubstituted cycloalkyl, tertiary butyloxycarbonyl acyl group, benzyloxy carbonyl acyl group, benzyl, substituted benzyl, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl;

X is oxygen, sulphur, carbon, NH, SO, SO ₂;

N is 1,2,3 or 4;

R ₄for hydroxyl, cyano group, carboxyl ,-COOR ₇,-CONR ₆r ₇, 1,2,4-oxadiazole, 1,3,4-oxadiazole, tetrazole, amino-1,3, the 4-oxadiazole of 5-, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl; R ₆, R ₇be selected from hydrogen separately respectively, C _1-3alkyl.

Prepare above-described compound, and the method for pharmacy acceptable salt or prodrug, it is characterized in that R in described formula I structure ₁, R ₃for hydrogen atom, X is the preparation method of the compound of Sauerstoffatom, comprises the steps:

1) compound that alkylated reaction obtains formula VIII structure is occurred under salt of wormwood condition to amino m-nitrophenol;

2) compound of formula VIII structure obtains corresponding hydrazinobenzene hydrochloride salt after diazotization reduction reaction, subsequently the compound that Fischer indoles ring closure reaction obtains formula IX structure occurs with pimelinketone;

3) there is the compound that aromatization obtains formula X structure in the compound of formula IX structure under the condition of DDQ, save this step then A ring be cyclohexene ring;

4) compound of formula X structure obtains corresponding aminocompound through catalytic hydrogenation, and obtains R in formula I structure after acyl chloride reaction ₃for hydrogen atom, X is the compound of Sauerstoffatom;

Wherein, A ring is phenyl ring or tetrahydrobenzene;

R is hydrogen atom, C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group, C ₂-C ₇alkenyl or alkynyl, replace or be not substituted cycloalkyl, tertiary butyloxycarbonyl acyl group, benzyloxy carbonyl acyl group, benzyl, substituted benzyl, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl;

N is 1,2,3 or 4;

Prepare above-described compound, and the method for pharmacy acceptable salt or prodrug, it is characterized in that R in described formula I structure ₁, R ₃for hydrogen atom, X is the preparation method of the compound of sulphur atom or sulfone, comprises the steps:

1) corresponding hydrazinobenzene hydrochloride salt is obtained after diazotization reduction reaction to amino m-nitrophenol, subsequently the compound that Fischer indoles ring closure reaction obtains formula XI structure occurs with pimelinketone;

2) compound of formula XI structure and trifluoromethanesulfanhydride anhydride generation esterification obtain the compound of formula XII structure;

3) there is the compound that aromatization obtains formula XIII structure in the compound of formula XII structure under the condition of DDQ, save this step then A ring be cyclohexene ring;

4) compound of formula XIII structure is at Pd ₂(dba) ₃the compound that linked reaction obtains formula XIV structure is there is with under the catalysis of XantPhos system;

5) compound of formula XIV structure uses vat powder to be amino by nitroreduction, obtains the compound of formula XV structure subsequently, be R in formula I structure through acylation reaction ₃for hydrogen atom, X is the compound of sulphur atom;

6) if X is sulfone, then the compound of formula XV structure can obtain R in formula I structure through oxidizing reaction ₃for hydrogen atom, X is the compound of sulfone.

Wherein, A ring is phenyl ring or tetrahydrobenzene;

X is S or SO ₂;

N is 1,2,3 or 4;

R ₄for hydroxyl, cyano group, carboxyl ,-COOR ₇,-CONR ₆r ₇, 1,2,4-oxadiazole, 1,3,4-oxadiazole, tetrazole, amino-1,3, the 4-oxadiazole of 5-, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl, R ₆, R ₇be selected from hydrogen separately respectively, C _1-3alkyl.

It should be noted that synthesis path described above is the preparation in order to illustrate the compounds of this invention, and preparation is never only limited to this, namely other synthetic methods are feasible equally, and described method refers to the synthetic method in skilled person's general knowledge.

If needed, the compounds of this invention can utilize methods known in the art to change their pharmacologically acceptable salt or the form of ester into.

Another object of the present invention is to provide the purposes of the compounds of this invention.

Inventor confirms through experiment, and the compounds of this invention has the activity of good suppression MEK.

Therefore, the present invention proposes compound of the present invention or the application in mek inhibitor prepared by its salt pharmaceutically accepted.Wherein, described mek inhibitor has the restraining effect to not activating MEK.

Accompanying drawing explanation

Fig. 1 is that checking compound is to the inhibiting experimental principle figure of non-phosphorylation MEK1.

Embodiment

Below by experiment, also the present invention will be further described in conjunction with the embodiments, it should be understood that these embodiments only for the object of illustration, never limit the scope of the invention.

The synthesis of embodiment 1,2-((4-hydroxyl-1-(4-tolysulfonyl amido)-9H-carbazole-3-base) sulfo-) acetic acid (MI-1)

The first step

In acetonitrile (20mL) solution of 9H-4-hydroxycarbazole (918.2mg, 5mmol), slowly add N-chlorosuccinimide (667.4mg, 5mmol), last 10 minutes.Reaction solution stirring at room temperature 1.5 hours, boils off solvent with Rotary Evaporators subsequently.Crude product silica gel column chromatography is separated (methylene dichloride/sherwood oil=1:5-1:2), and obtain product 3-chloro-4-hydroxyl carbazole, white solid (518.2mg), productive rate is 47.6%.

¹H NMR(400MHz,DMSO-d ₆):δ11.31(s,1H,NH),9.78(s,1H,OH),8.21(d,J＝8Hz,1H,ArH),7.46(d,J＝8Hz,1H,ArH),7.36(td,J＝1.16Hz J＝8Hz,1H,ArH),7.30(d,J＝8Hz,1H,ArH),7.16(td,J＝0.96Hz J＝8Hz,1H,ArH),7.00(d,J＝8Hz,1H,ArH).

Second step

Water (85mL) solution of nitroso-group Potassium Persulphate (1.4430g) and potassium primary phosphate (84.5mg) is added drop-wise in acetone (85mL) solution of 3-chloro-4-hydroxyl carbazole (410mg, 1.88mmol).Reaction solution stirring at room temperature 1.5 hours, then uses Büchner funnel suction filtration, a small amount of water washing of filter cake, infrared lower oven dry.Obtain the chloro-carbazole-Isosorbide-5-Nitrae-diketone of 3-, be dark red solid (413.3mg), productive rate is 94.7%.

¹H NMR(400MHz,DMSO-d ₆):δ13.11(s,1H,NH),8.05(d,J＝8Hz,1H,ArH),7.57(d,J＝8Hz,1H,ArH),7.43(td,J＝0.96Hz J＝8Hz,1H,ArH),7.37(td,J＝0.96Hz J＝8Hz,1H,ArH),7.18(s,1H,ArH).

3rd step

To the chloro-carbazole-1 of 3-under condition of ice bath, 4-diketone (116mg, 0.5mmol) with p-methylphenyl sulphonylamine (85.6mg, dichloromethane solution (the 1mL of titanium tetrachloride is dripped in tetrahydrofuran solution (6mL) 0.5mmol), 1M), and triethylamine (0.16mL).Mixed solution is reacting by heating 15 minutes (150 watts in microwave reaction instrument, 65 degrees Celsius), subsequently with short silicagel column filtering insoluble solid particle, (ethyl acetate/petroleum ether=5:1-1:1) is separated with silicagel column after filtrate mixes sample, obtain coffee-like solid (132.8mg), productive rate is 69.0%.

¹H NMR(400MHz,DMSO-d ₆):δ12.87(s,1H,aromatic NH),8.11(s,1H,ArH),8.00(m,3H,ArH),7.54(d,J＝8Hz,3H,ArH),7.41(t,J＝8Hz,1H,ArH),7.33(t,J＝8Hz,1H,ArH),2.46(s,3H,CH ₃).

4th step

Previous step product (77mg, 0.2mmol) is dissolved in tetrahydrofuran (THF) (6mL), adds Thiovanic acid (20.8uL, 0.3mmol) and pyridine (24.2uL, 0.3mmol).Reaction solution stirring at room temperature is concentrated solvent after 40 minutes.Mixture ethyl acetate (50mL) is redissolved, and is transferred to 250mL separating funnel, washes away pyridine with sulfuric acid one hydrogen potassium solution (0.5M, 20mL X 2).Take a policy powder (204.7mg, 0.5mmol) and water (50mL) subsequently, and fierce jolting is until ethyl acetate layer becomes light yellow.Reject aqueous phase, uses water (50mL X 2) and saturated aqueous common salt (50mL) to wash organic phase respectively, then uses anhydrous sodium sulfate drying.Crude product is separated through silicagel column and obtains yellow solid (21mg), and productive rate is 21.3%.

¹H NMR(400MHz,DMSO-d ₆):δ13.33(br s,1H,COOH),11.06(s,1H,aromaticNH),9.48(s,1H,SO ₂NH),8.17(d,J＝8Hz,1H,ArH),7.57-7.51(m,3H,ArH),7.34-7.30(m,3H,ArH),7.14(t,J＝8Hz,1H,ArH),6.65(s,1H,ArH),3.13(s,2H,CH ₂),2.36(s,3H,CH ₃).

¹³C NMR(100MHz,DMSO-d ₆)δ172.23,153.62,143.58,139.61,139.44,136.81,130.76,129.88,127.67,125.57,122.51,122.21,119.79,113.36,112.68,111.69,106.30,21.44.

ESI-MS(C ₂₁H ₁₈N ₂O ₅S ₂):m/z 441.0593(M-H ⁺)；ESI-HRMS(C ₂₁H ₁₈N ₂O ₅S ₂):m/z441.05734(M-H ⁺).

The synthesis of embodiment 2,2-((the bromo-4-hydroxyl of 6--1-(4-tolysulfonyl amido)-9H-carbazole-3-base) sulfo-) acetic acid (MI-2)

The first step

The hydrochloride (222.8mg, 1mmol) of 4-bromophenyl-hydrazine and hydroresorcinol (125.9mg, 1.12mmol) are dissolved in trifluoroacetic acid (1.5mL).Mixture uses microwave reaction instrument reacting by heating 10 minutes (140 degrees Celsius, 50 watts).Revolve and steam except desolventizing, solid redissolves in ethyl acetate (60mL), washes twice (50mL) remove trifluoroacetic acid, reject aqueous phase with saturated sodium bicarbonate solution.Revolve and steam except desolventizing, gained solid is making beating in methylene dichloride (20mL), subsequent filtration.The a small amount of washed with dichloromethane of filter cake, obtains greyish-green solid (118.5mg), and productive rate is 44.9%.

¹H NMR(400MHz,DMSO-d ₆):δ12.07(s,1H,NH),8.05(d,J＝1.92Hz,1H,ArH),7.38(d,J＝8.52Hz,1H,ArH),7.30(dd,J＝1.96Hz J＝8.52Hz,1H,ArH),2.97(t,2H,ArC H ₂),2.44(t,2H,COC H ₂),2.12(p,2H,CH ₂C H ₂CH ₂).

Second step

Anhydrous cupric chloride (2.689g, 20mmol) is added, Lithium chloride (anhydrous) (512mg, 12mmol) in DMF (12mL) solution of 6-bromine tetrahydro carbazole-4-ketone (1.055g, 4mmol).Mixed solution 105 degrees Centigrade 1 hour, subsequent filtration removing inorganic salt.Filtrate, with the dilution of 100mL water, is then extracted with ethyl acetate (50mLX 3).Organic phase after merging is with saturated common salt water washing (50mL X 3), and anhydrous sodium sulfate drying, obtains brown oil after concentrated.

This brown oil is dissolved in DMF (15mL), adds Lithium chloride (anhydrous) (640.2mg, 15mmol) and Quilonum Retard (1.1083g, 15mmol).Mixture is heated to 150 degrees Celsius of reactions 4 hours under nitrogen protection condition.Then cool to room temperature, use 100mL diluted ethyl acetate, cross and filter inorganic salt, then use dilute hydrochloric acid (2N, 50mL X 2) and saturated common salt water washing, use anhydrous sodium sulfate drying subsequently.Crude product is separated through column chromatography (ethyl acetate/petroleum ether=1:5) and obtains brown solid (403mg), and productive rate is 34%.

¹H NMR(400MHz,DMSO-d ₆):δ11.29(s,1H,NH),10.18(s,1H,OH),8.23(d,J＝1.32Hz,1H,ArH),7.44(dd,J＝1.76Hz J＝8.56Hz,1H,ArH),7.39(d,J＝8.52Hz,1H,ArH),7.20(t,1H,ArH),6.93(d,J＝8.04Hz,1H,ArH),6.59(d,J＝7.76Hz,1H,ArH).

¹³C NMR(100MHz,DMSO-d ₆):δ154.19,142.53,138.20,127.81,127.01,124.52,124.51,112.59,110.69,110.67,104.79,102.55.

3rd step

In acetonitrile (4mL) solution of 9H-4-hydroxyl-6-bromine carbazole (130.5mg, 0.5mmol l), slowly add N-chlorosuccinimide (66.3mg, 0.5mmol), last 10 minutes.Reaction solution stirring at room temperature 1.5 hours, concentrated solvent subsequently.Crude product silica gel column chromatography is separated (methylene dichloride/sherwood oil=1:3), and obtain product 3-chloro-4-hydroxyl-6-bromine carbazole, silvery white solid (77.2mg), productive rate is 52.3%.

¹H NMR(400MHz,DMSO-d ₆):δ11.51(s,1H,NH),10.02(s,1H,OH),8.29(d,J＝1.24Hz,1H,ArH),7.50(dd,J＝1.68Hz J＝8.60Hz,1H,ArH),7.44(d,J＝8.60Hz,1H,ArH),7.35(d,J＝8.56Hz,1H,ArH),7.02(d,J＝8.56Hz,1H,ArH).

¹³C NMR(100MHz,DMSO-d ₆):δ148.83,140.86,138.64,127.89,127.80,124.70,123.85,113.11,112.55,111.07,109.69,104.46.

4th step

By 3-chloro-4-hydroxyl-6-bromine carbazole (213.5mg, acetone (33mL) solution 0.72mmol) is added drop-wise to nitroso-group Potassium Persulphate (563.5mg, 1.224mmol) with in water (33mL) solution of potassium primary phosphate (34.6mg, 0.24mmol).Reaction solution stirring at room temperature 3 hours, then uses Büchner funnel suction filtration, a small amount of water washing of filter cake, infrared lower oven dry.Obtain the chloro-6-bromine carbazole-Isosorbide-5-Nitrae-diketone of 3-with acetone recrystallization, be dark red solid (75.0mg), productive rate is 33.5%.

¹H NMR(400MHz,DMSO-d ₆):δ13.25(s,1H,carbazoleNH),8.09(d,J＝8.0Hz,1H,ArH),7.50(m,2H,ArH),7.18(d,J＝8.0Hz,1H,ArH).

5th step

To 3-chloro-6-bromine carbazole-1,4-diketone (64mg, 0.21mmol) with p-methylphenyl sulphonylamine (35.2mg, dichloromethane solution (the 0.21mL of titanium tetrachloride is dripped in tetrahydrofuran (THF) (2.5mL) solution 0.21mmol), 1M), and triethylamine (66uL).Mixed solution is reacting by heating 15 minutes (150 watts in microwave reaction instrument, 65 degrees Celsius), subsequently with short silicagel column filtering insoluble solid particle, after concentrated filtrate, gained solid with ethyl acetate recrystallization obtains coffee-like solid (56.9mg), and productive rate is 59.6%.

6th step

Previous step product (49mg, 0.1mmol) is dissolved in tetrahydrofuran (THF) (3mL), adds Thiovanic acid (11uL, 0.15mmol) and pyridine (12.5uL, 0.15mmol).Reaction solution stirring at room temperature is concentrated solvent after 30 minutes.Mixture ethyl acetate (20mL) is redissolved, and is transferred to 100mL separating funnel, washes away pyridine with sulfuric acid one hydrogen potassium solution (0.5M, 20mL X 2).Take a policy powder (102.4mg, 0.5mmol) and water (20mL) subsequently, and fierce jolting is until ethyl acetate layer becomes light yellow.Reject aqueous phase, uses water (20mL X 2) and saturated aqueous common salt (20mL) to wash organic phase respectively, then uses anhydrous sodium sulfate drying.Crude product is separated through silicagel column (ethanol/methylene=1:8) and obtains brown solid (9.7mg), and productive rate is 18.6%.

¹H NMR(400MHz,DMSO-d ₆):δ11.17(s,1H,carbazoleNH),9.46(s,1H,NHSO ₂),8.28(d,1H,ArH),7.54(d,J＝7.2Hz,2H,ArH),7.44(m,2H,ArH),7.33(d,J＝7.2Hz,2H,ArH),6.60(s,1H,ArH),3.02(s,2H,SCH ₂),2.36(s,3H,CH ₃).

ESI-MS(C21H17BrN2O5S2):m/z 518.9657520.9666(M-H+)；ESI-HRMS(C21H17BrN2O5S2):m/z 518.96895(M-H+).

The synthesis of embodiment 3, ethyl 2-((1-(4-p-methylphenyl sulphonylamine base)-9H-carbazole-3-base) oxo) acetic ester (MI-3)

The synthesis of the first step intermediate compound I M1

Add in 100mL round-bottomed bottle amino m-nitrophenol (4.9952g, 32.4mmol), salt of wormwood (4.9346g, 35.6mmol) and 60mL acetone.Drip ethyl bromoacetate (3.8mL, 34.0mmol) subsequently.Reaction mixture stirred overnight at room temperature, filtering inorganic salt.Filtrate concentrates to obtain yellow solid, and gained solid is making beating in ethyl acetate and sherwood oil mixed solution (1:3,50mL), and use Büchner funnel suction filtration subsequently, obtain yellow solid (5.8554g), productive rate is 75.2%.

¹H NMR(400MHz,DMSO-d ₆):δ7.37(d,J＝2.24Hz,1H,ArH),7.27(s,2H,NH ₂),7.22(dd,J＝2.68Hz J＝9.24Hz,1H,ArH),7.01(d,J＝9.24Hz,1H,ArH),4.74(s,2H,OC H ₂CO),4.17(q,J＝7.08Hz,2H,COC H ₂CH ₃),1.21(t,J＝7.12Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.09,147.86,142.71,129.45,127.71,121.18,107.47,65.97,61.11,14.50.

The synthesis of second step intermediate compound I M2

Intermediate compound I M1 (5.8554g, 24.4mmol) is dissolved in concentrated hydrochloric acid (40mL), is cooled to-10 degrees Celsius.Water (3mL) solution dripping Sodium Nitrite (2.027g, 29.28mmol) carries out diazotization.Reaction drips tin protochloride (11.04g, 48.8mmol) concentrated hydrochloric acid (10mL) solution after half an hour carries out reduction reaction, completes after dripping and keeps-10 degrees Celsius of reactions 1 hour, be then warmed up to room temperature, at room temperature stir and spend the night.

By above-mentioned concentrated hydrochloric acid solution ethanol (50mL) dilution, slowly add pimelinketone (2.5mL, 24.4mmol).Mixed solution, 90 degrees Celsius of backflows 3 hours, is then down to room temperature.Use Büchner funnel suction filtration, filter cake a small amount of water washing three times, the infrared lower oven dry of gained solid.Obtain red solid (3.2376g), productive rate is 41.7%.

¹H NMR(400MHz,CDCl ₃):δ9.34(s,1H,indole NH),7.66(s,1H,ArH),7.38(s,1H,ArH),4.72(s,2H,OC H ₂CO),4.32(q,J＝7.12Hz,2H,COC H ₂CH ₃),2.79(t,2H,1-CH ₂),2.67(t,2H,4-CH ₂),1.93(m,4H,2,3-CH ₂CH ₂)1.34(t,J＝7.08Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,CDCl ₃):δ168.83,150.81,138.51,132.31,131.41,125.74,112.82,111.13,105.03,66.90,61.45,23.17,22.86,22.80,20.56,14.18.

The synthesis of the 3rd step intermediate compound I M3

Using intermediate compound I M2 (3.2376g, 10.17mmol) and DDQ at toluene (50mL) as reflux under the condition of solvent 2 hours, cool to room temperature subsequently.Mixture 100mL diluted ethyl acetate, transfers to 250mL separating funnel, and organic phase saturated sodium bicarbonate solution washing (100mL X 5), until aqueous phase is colourless.Reject aqueous phase, organic phase anhydrous sodium sulfate drying, concentrates and obtains red brown solid (3.4166g), quantitative reaction.

¹H NMR(400MHz,DMSO-d ₆):δ11.97(s,1H,NH),8.37(s,1H,ArH),8.23(d,J＝7.76Hz,1H,ArH),7.86(s,1H,ArH),7.71(d,J＝8.16Hz,1H,ArH),7.51(t,1H,ArH),7.27(t,1H,ArH),4.98(s,2H,OC H ₂CO),4.21(q,J＝7.08Hz,2H,COC H ₂CH ₃),1.24(t,J＝7.16Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.15,150.61,141.93,131.35,129.40,128.26,128.06,121.81,121.46,120.69,116.03,113.05,108.62,66.58,61.18,14.51.

The synthesis of the 4th step intermediate compound I M4

Previous step aromatization products IM3 (315.1mg, 1mmol) is dissolved in methyl alcohol (25mL), adds palladium carbon (20%, 65.7mg).Mixture is stirred overnight at room temperature in hydrogenation instrument (40psi).Filtering palladium carbon, is directly used in next step after concentrated filtrate.

5th step

Previous step gained intermediate compound I M4 is dissolved in pyridine (10mL), adds p-methyl benzene sulfonic chloride (263.1mg, 1.38mmol) in batches.Mixture stirred overnight at room temperature.Steamed by solvent pyridine with surge pump, crude product silica gel column chromatography is separated (sherwood oil/acetone=3:1) and obtains white solid (221.1mg), and productive rate is 50.4%.

¹H NMR(400MHz,DMSO-d ₆):δ10.75(s,1H,carbazoleNH),10.00(s,1H,NHSO ₂),8.02(d,J＝7.80Hz,1H,ArH),7.68(d,J＝8.24Hz,2H,ArH),7.54(d,J＝8.16Hz,1H,ArH),7.49(d,J＝2.24Hz,1H,ArH),7.37(td,1H,ArH),7.33(d,J＝8.08Hz,2H,ArH),7.12(t,1H,ArH),6.68(d,J＝2.32Hz,1H,ArH),4.70(s,2H,OC H ₂CO),4.17(q,J＝7.12Hz,2H,COC H ₂CH ₃),2.32(s,3H,PhC H ₃),1.22(t,J＝7.08Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.32,151.55,143.85,140.43,137.08,130.11,129.43,127.34,126.36,124.27,122.71,121.93,120.78,119.00,112.02,109.07,102.06,66.16,61.00,21.40,14.51.

ESI-MS(C23H22N2O5S):m/z 437.1193(M-H+)；ESI-HRMS(C23H22N2O5S):m/z 437.11767(M-H+).

The synthesis of embodiment 4, ethyl 2-((1-(4-acetylaminobenzene sulfonamide base)-9H-carbazole-3-base) oxo) acetic ester (MI-4)

First to fourth step

With embodiment 3

5th step

Previous step gained intermediate compound I M4 is dissolved in pyridine (5mL), adds p-acetaminobenzenesulfonyl chloride (322.5mg, 1.38mmol) in batches.Mixture stirred overnight at room temperature.Steamed by solvent pyridine with surge pump, crude product silica gel column chromatography is separated (sherwood oil/acetone=1:1) and obtains white solid (103.9mg), and productive rate is 21.6%.

¹H NMR(400MHz,DMSO-d ₆):δ10.73(s,1H,carbazoleNH),10.26(s,1H,N HCOCH ₃),9.93(s,1H,NHSO ₂),8.02(d,J＝7.76Hz,1H,ArH),7.70(m,4H,ArH),7.54(d,J＝8.12Hz,1H,ArH),7.49(s,1H,ArH),7.37(t,1H,ArH),7.12(t,1H,ArH),6.68(s,1H,ArH),4.71(s,2H,OCH ₂CO),4.17(q,J＝7.08Hz,2H,COC H ₂CH ₃),2.04(s,3H,NHCOC H ₃),1.22(t,J＝7.08Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.45,169.34,151.55,143.70,140.44,133.34,129.53,128.56,126.35,124.26,122.71,121.95,120.77,118.98,118.90,112.02,109.25,102.06,66.15,60.99,24.56,14.51.

ESI-MS(C ₂₄H ₂₃N ₃O ₆S):m/z 480.1262(M-H ⁺)；ESI-HRMS(C ₂₄H ₂₃N ₃O ₆S):m/z480.12348(M-H ⁺).

The synthesis of embodiment 5,2-((1-(4-p-methylphenyl sulphonylamine base)-9H-carbazole-3-base) oxo) acetic acid (MI-5)

Sodium hydroxide (20mg, 0.5mmol) is dissolved in 1mL water, is added drop-wise in tetrahydrofuran (THF) (1mL) solution of MI-3 (76.5mg, 0.17mmol).Mixture uses 20mL diluted ethyl acetate in stirring at room temperature after 1.5 hours, extracts three times with saturated sodium bicarbonate solution, each 20mL.Merge aqueous phase, adjust pH to pH=1 gradually with 2M dilute hydrochloric acid solution, white solid is slowly precipitated out.Use Büchner funnel suction filtration, filter white solid in infrared lower oven dry, be weighed as 58.0mg, yield is 81%.

¹H NMR(400MHz,DMSO-d ₆):δ12.96(br s,1H,COOH),10.75(s,1H,carbazoleNH),10.02(br s,1H,NHSO ₂),8.02(d,J＝7.80Hz,1H,ArH),7.70(d,J＝7.92Hz,2H,ArH),7.54(d,J＝8.12Hz,1H,ArH),7.47(s,1H,ArH),7.37(t,1H,ArH),7.33(d,J＝8.08Hz,2H,ArH),7.11(t,1H,ArH),6.70(s,1H,ArH),4.61(s,2H,CH ₂),2.32(s,3H,CH ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ170.84,151.74,143.86,140.42,137.10,130.12,129.30,127.36,126.31,124.24,121.96,120.76,118.97,112.00,109.07,101.75,65.94,21.41.

ESI-HRMS(C ₂₁H ₁₈N ₂O ₅S):m/z 409.08637(M-H ⁺).

The synthesis of embodiment 6,2-((1-(4-acetylaminobenzene sulfonamide base)-9H-carbazole-3-base) oxo) acetic acid (MI-6)

Sodium hydroxide (15.2mg, 0.4mmol) is dissolved in 1mL water, is added drop-wise in tetrahydrofuran (THF) (1mL) solution of MI-4 (48.3mg, 0.1mmol).Mixture uses 20mL diluted ethyl acetate in stirring at room temperature after 1 hour, extracts three times with saturated sodium bicarbonate solution, each 20mL.Merge aqueous phase, adjust pH to pH=1 gradually with 2M dilute hydrochloric acid solution, white solid is slowly precipitated out.Use Büchner funnel suction filtration, filter white solid in infrared lower oven dry, be weighed as 20.7mg, yield is 45.5%.

¹H NMR(400MHz,DMSO-d ₆):δ12.95(s,1H,COOH),10.71(s,1H,carbazoleNH),10.25(s,1H,N HCOCH ₃),9.94(s,1H,SO ₂NH),8.02(d,J＝6.68Hz,1H,ArH),7.71(d,J＝7.60Hz,4H,ArH),7.53(d,J＝7.88Hz,1H,ArH),7.46(s,1H,ArH),7.36(t,1H,ArH),7.11(t,1H,ArH),6.71(s,1H,ArH),4.62(s,2H,OCH ₂),2.04(s,3H,COCH ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ170.79,169.45,151.74,143.69,140.42,133.40,129.33,128.57,126.29,124.22,122.74,122.00,120.75,118.96,111.99,109.23,101.69,65.93,24.56.

ESI-HRMS(C ₂₂H ₁₉N ₃O ₆S):m/z 452.09218(M-H ⁺).

The synthesis of embodiment 7,2-((1-(4-p-methylphenyl sulphonylamine base)-9H-carbazole-3-base) oxo) ethanamide (MI-7)

MI-3 (440mg, 1mmol) is dissolved in 18mL strong aqua, mixture stirring at room temperature 3 hours, solid dissolves completely, and solution is light yellow.Reacted rear concentrated solvent except deammoniation, solid, in infrared lower oven dry, is weighed as 389.9mg, and yield is 94.9%.

¹H NMR(400MHz,DMSO-d ₆):δ10.74(s,1H,carbazole N H),10.00(s,1H,SO ₂N H),8.01(d,J＝7.76Hz,1H,ArH),7.71(d,J＝8.08Hz,2H,ArH),7.54(d,J＝8.08Hz,1H,ArH),7.49(d,J＝1.64Hz,1H,ArH),7.41-7.31(m,4H,ArH),7.12(t,1H,ArH),6.84(d,J＝1.84Hz,1H,ArH),4.39(s,2H,CH ₂),2.30(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ170.57,151.74,143.87,140.37,137.05,130.12,129.19,127.39,126.33,124.19,122.72,122.02,120.70,119.03,112.02,109.09,101.85,68.20,21.41.

ESI-HRMS(C ₂₁H ₁₉N ₃O ₄S):m/z 408.10235(M-H ⁺).

The synthesis of embodiment 8, N-(3-((1H-tetrazole-5-base) methoxyl group)-9H-carbazole-1-base)-4-p-methylphenyl sulphonylamine (MI-8)

Under argon shield condition, MI-7 (271.8mg, 0.66mmol); sodiumazide (651.6mg; 9.96mmol), silicon tetrachloride (0.38mL, 3.32mmol) is heated to 80 degrees Celsius of reactions 14 hours in anhydrous acetonitrile (6mL).Room temperature is cooled to after having reacted, with 50mL methanol dilution reaction solution, filtering inorganic salt.Be separated (ethanol/methylene=1:30-1:20) with silica gel column chromatography after filtrate is concentrated, obtain white solid 179.2mg, yield is 62.1%.

¹H NMR(400MHz,DMSO-d ₆):δ10.79(s,1H,carbazole N H),10.05(s,1H,SO ₂N H),8.04(d,J＝7.80Hz,1H,ArH),7.68(d,3H,ArH),7.56(d,J＝8.16Hz,1H,ArH),7.38(t,1H,ArH),7.31(d,J＝8.08Hz,2H,ArH),7.14(t,1H,ArH),6.83(d,J＝2.00Hz,1H,ArH),5.46(s,2H,CH ₂),2.30(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ151.31,143.91,140.39,136.96,130.14,129.41,127.32,126.47,124.25,122.69,122.10,120.77,119.13,112.09,109.20,102.31,60.87,21.41.

ESI-MS(C ₂₁H ₁₈N ₆O ₃S):m/z 433.28(M-H ⁺)；ESI-HRMS(C ₂₁H ₁₈N ₆O ₃S):m/z434.11556(M ⁺)473.08103(M+K ⁺).

The synthesis of embodiment 9, N-(3-((amino-1,3, the 4-oxadiazole of 5--2-base) methoxyl group)-9H-carbazole-1-base)-4-p-methylphenyl sulphonylamine (MI-9)

The synthesis of the first step intermediate compound I M5

MI-3 (943.4,2.15mmol) and hydrazine hydrate (1.05mL, 50wt%) are refluxed one day in dehydrated alcohol (13mL), solid dissolves gradually, and solution is clarified.Room temperature is cooled to, by the solid filtering of precipitation out, with the then infrared lower oven dry of a small amount of washing with alcohol after reaction terminates.Obtain white solid 758.2mg, yield is 83.0%.

¹H NMR(400MHz,DMSO-d ₆):δ10.73(s,1H,carbazole NH),10.01(s,1H,SO ₂NH),9.39(s,1H,CON HNH ₂),8.01(d,J＝7.72Hz,1H,ArH),7.71(d,J＝7.96Hz,2H,ArH),7.54(d,J＝8.12Hz,1H,ArH),7.49(s,1H,ArH),7.37(t,1H,ArH),7.33(d,J＝7.92Hz,2H,ArH),7.12(t,1H,ArH),6.84(s,1H,ArH),4.45(s,2H,CH ₂),4.36(s,2H,CONHN H ₂),2.31(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ167.17,151.81,143.86,140.35,137.07,130.13,129.15,127.38,126.32,124.14,122.73,122.02,120.70,119.01,112.03,109.22,101.61,67.72,21.41.

Second step

Intermediate compound I M5 (422.1mg, 1mmol) obtained in the previous step and cyanogen bromide (121.0mg, 1.2mmol) are refluxed 3 hours in 10mL ethanol.Be separated (acetone/methylene dichloride=1:4-1:2) with silica gel column chromatography after concentrated solvent, obtain white solid 102mg, yield is 22.8%.

¹H NMR(400MHz,DMSO-d ₆):δ10.79(s,1H,carbazole N H),10.02(s,1H,SO ₂N H),8.02(d,J＝7.20Hz,1H,ArH),7.68(t,3H,ArH),7.55(d,J＝7.64Hz,1H,ArH),7.38(t,1H,ArH),7.32(d,J＝7.08Hz,2H,ArH),7.20(s,2H,NH ₂),7.14(t,1H,ArH),6.74(s,1H,ArH),5.11(s,2H,CH ₂),2.30(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ164.90,155.93,151.26,143.90,140.41,136.95,130.14,129.51,127.37,126.45,124.26,122.69,122.05,120.74,119.11,112.08,109.34,102.54,61.17,21.41.

ESI-HRMS(C ₂₂H ₁₉N ₅O ₄S):m/z 450.12305(M+H ⁺)472.10488(M+Na ⁺)488.07883(M+K ⁺).

The synthesis of embodiment 10,4-methyl-N-(3-((3-methyl isophthalic acid, 2,4-oxadiazole-5-base) methoxyl group)-9H-carbazole-1-base) benzsulfamide (MI-10)

The synthesis of the first step intermediate compound I M6

In the tetrahydrofuran solution (12mL) of IM3 (1.255g, 4mmol), drip the aqueous solution (12mL) of lithium hydroxide (338.6mg, 8mmol), dropwise rear stirring at room temperature 4.5 hours.React rear reaction solution 80mL diluted ethyl acetate, extracted three times (each 80mL) with saturated sodium bicarbonate solution.Merge aqueous phase, slowly regulate pH to 1 with the dilute hydrochloric acid of 10%, the yellow solid Büchner funnel suction filtration of precipitation, a small amount of water washing, then infrared lower oven dry.Be weighed as 1.14g, yield is 99.8%.

¹H NMR(400MHz,DMSO-d ₆):δ13.11(br s,1H,COOH),11.97(s,1H,NH),8.37(s,1H,ArH),8.24(d,J＝7.68Hz,1H,ArH),7.85(s,1H,ArH),7.71(d,J＝8.04Hz,1H,ArH),7.51(t,1H,ArH),7.27(t,1H,ArH),4.89(s,2H,C H ₂COOH).

¹³C NMR(100MHz,DMSO-d ₆):δ170.65,150.84,141.93,131.37,129.31,128.24,128.05,121.82,121.48,120.68,115.95,113.04,108.46,66.35.

The synthesis of second step intermediate compound I M7

By intermediate compound I M6 (167.0mg, 0.58mmol), HOBt (95.7mg, 0.7mmol) and EDCI (129.6mg, 0.67mmol) be stirring at room temperature half an hour in anhydrous acetonitrile (2mL) first.Drip the acetonitrile solution (1.5mL) of acetyl amidoxime (57.3mg, 0.61mmol) subsequently, then mixture flows through night next time in heating 90 degrees celsius.Concentrated solvent, solid silica gel column chromatography is separated (petrol ether/ethyl acetate=3:1) and obtains orange solids 101.8mg, and yield is 53.8%.

¹H NMR(400MHz,DMSO-d ₆):δ12.04(s,1H,NH),8.49(d,J＝1.68Hz,1H,ArH),8.24(d,J＝7.80Hz,1H,ArH),8.01(d,J＝1.68Hz,1H,ArH),7.72(d,J＝8.16Hz,1H,ArH),7.52(t,1H,ArH),7.28(t,1H,ArH),5.70(s,2H,OCH ₂),2.38(s,3H,CH ₃).

¹³C NMR(100MHz,CDCl ₃):δ174.09,167.61,150.33,140.60,131.25,130.04,128.23,128.20,121.76,121.09,120.79,115.47,111.72,108.31,62.55,11.56.

3rd step

By intermediate compound I M7 (153.7mg; 0.474mmol) be dissolved in the mixed solution (1:1 of methyl alcohol and methylene dichloride; 30mL); with slowly dripping V-Brite B (1.03g half an hour under nitrogen protection condition; 5.688mmol) and the aqueous solution (4mL) of salt of wormwood (814.0mg, 5.688mmol).Dropwise rear stirring at room temperature two hours, subsequently filtering inorganic salt, after filtrate is concentrated, be directly used in next step.

Reduzate is dissolved in pyridine (10mL), slowly adds Tosyl chloride (128.8mg, 0.65mmol).Mixture stirred overnight at room temperature.Steam except pyridine with surge pump, crude product is separated (sherwood oil/acetone=3:1) through silica gel column chromatography and obtains white solid 29.2mg, and yield is 13.7%.

¹H NMR(400MHz,DMSO-d ₆):δ10.79(s,1H,carbazoleNH),10.02(s,1H,NHSO ₂),8.02(d,J＝7.76Hz,1H,ArH),7.67(d,J＝8.12Hz,2H,ArH),7.65(s,1H,ArH),7.55(d,J＝8.24Hz,1H,ArH),7.38(t,1H,ArH),7.32(d,J＝7.96Hz,2H,ArH),7.13(t,1H,ArH),6.75(s,1H,ArH),5.42(s,2H,CH ₂),2.36(s,3H,oxadiazoleC H ₃),2.31(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ175.59,167.51,151.20,143.90,140.45,136.99,130.12,129.67,127.34,126.49,124.27,122.67,122.09,120.78,119.13,112.09,109.21,102.48,62.19,21.41,11.55.

ESI-MS(C ₂₃H ₂₀N ₄O ₄S):m/z 447.1143(M-H ⁺)；ESI-HRMS(C ₂₃H ₂₀N ₄O ₄S):m/z447.11215(M-H ⁺).

The synthesis of embodiment 11, N-(4-(N-(3-((3-methyl isophthalic acid, 2,4-oxadiazole-5-base) methoxyl group)-9H-carbazole-1-base) amino-sulfonyl) phenyl) ethanamide (MI-11)

By intermediate compound I M7 (165.4mg; 0.5mmol) be dissolved in the mixed solution (1:1 of methyl alcohol and methylene dichloride; 20mL); with slowly dripping V-Brite B (1.06g half an hour under nitrogen protection condition; 6mmol) and the aqueous solution (4mL) of salt of wormwood (828.3mg, 6mmol).Dropwise rear stirring at room temperature two hours, subsequently concentrated solvent, crude product 50mL water and 30mL methylene dichloride separatory, aqueous phase uses dichloromethane extraction twice again, each 30mL.Use anhydrous sodium sulfate drying after merging organic phase, after organic phase is concentrated, be directly used in next step.

Reduzate is dissolved in pyridine (5mL), slowly adds p-acetaminobenzenesulfonyl chloride (199mg, 0.85mmol).Mixture stirred overnight at room temperature.Steam except pyridine with surge pump, crude product is separated (sherwood oil/acetone=3:2) through silica gel column chromatography and obtains white solid 78.3mg, and yield is 31.2%.

¹H NMR(400MHz,DMSO-d ₆):δ10.78(s,1H,carbazole NH),10.26(s,1H,N HCOCH ₃),9.97(s,1H,SO ₂NH),8.03(d,J＝7.68Hz,1H,ArH),7.73-7.65(m,5H,ArH),7.55(d,J＝8.04Hz,1H,ArH),7.38(t,J＝7.48Hz,1H,ArH),7.13(t,J＝7.20Hz,1H,ArH),6.77(s,1H,ArH),5.44(s,2H,CH ₂),2.37(s,3H,oxadiazoleCH ₃),2.04(s,3H,NHCOC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ175.60,169.45,167.51,151.21,143.73,140.46,133.28,129.75,128.57,126.47,124.26,122.68,122.12,120.79,119.11,118.92,112.08,109.37,102.54,62.23,24.56,11.54.

ESI-HRMS(C ₂₄H ₂₁N ₅O ₅S):m/z 490.11906(M-H ⁺).

The synthesis of embodiment 12,4-methyl-N-(3-((5-methyl isophthalic acid, 3,4-oxadiazole-2-base) methoxyl group)-9H-carbazole-1-base) benzsulfamide (MI-12)

The synthesis of the first step intermediate compound I M8

By IM6 (561.7mg, 1.96mmol), acethydrazide (291.4mg, 4mmol), HOBt (400.6mg, 3mmol) and EDCI (565.4mg, 3mmol) is dissolved in 8mL DMF.Mixture stirring at room temperature 12 hours, has reacted rear dropping 40mL water and product is separated out.Use Büchner funnel suction filtration, filter cake infrared drying after a small amount of water washing.Obtain red solid 502mg, yield 74.7%.Be directly used in next step.

The synthesis of second step intermediate compound I M9

IM8 (173.2mg, 0.5mmol) is dissolved in phosphorus oxychloride (2mL), reacts and carry out under tube sealing condition, be heated to 100 degrees Celsius of reactions 1 hour.Be cooled to room temperature subsequently, concentrated solvent, add saturated sodium bicarbonate solution and adjust pH for alkalescence, use dichloromethane extraction (50mL x 2) subsequently.Organic phase anhydrous sodium sulfate drying, after concentrated solvent, crude product silica gel column chromatography is separated (petrol ether/ethyl acetate=3:1) and obtains yellow solid 107.9mg, and yield is 65.8%.

¹H NMR(400MHz,DMSO-d ₆):δ12.02(s,1H,NH),8.47(s,1H,ArH),8.23(d,J＝7.76Hz,1H,ArH),8.02(s,1H,ArH),7.72(d,J＝8.08Hz,1H,ArH),7.52(t,1H,ArH),7.28(t,1H,ArH),5.57(s,2H,CH ₂),2.55(s,3H,CH ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ165.30,162.78,150.18,141.98,131.40,129.69,128.33,128.19,121.78,121.46,120.80,116.53,113.11,108.95,61.49,10.97.

3rd step

Previous step product IM9 (91.2mg, 0.28mmol) is dissolved in methyl alcohol (25mL), adds palladium carbon (10%, 32.8mg).Mixture is stirred overnight at room temperature in hydrogenation instrument (40psi).Filtering palladium carbon, is directly used in next step after filtrate is concentrated.

Reduzate is dissolved in pyridine (5mL), adds p-methyl benzene sulfonic chloride (79.8mg, 0.39mmol) in batches.Mixture stirred overnight at room temperature.Steamed by solvent pyridine with surge pump, crude product silica gel column chromatography is separated (sherwood oil/acetone=2:1-1:1) and obtains white solid (34.6mg), and productive rate is 27.4%.

¹H NMR(400MHz,DMSO-d ₆):δ10.77(s,1H,carbazoleNH),10.13(s,1H,NHSO ₂),8.02(d,J＝7.52Hz,1H,ArH),7.68(d,J＝7.52Hz,2H,ArH),7.62(s,1H,ArH),7.55(d,J＝7.96Hz,1H,ArH),7.37(t,1H,ArH),7.31(d,J＝7.52Hz,2H,ArH),7.13(t,1H,ArH),6.77(s,1H,ArH),5.30(s,2H,CH ₂),2.53(s,3H,oxadiazoleC H ₃),2.30(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ165.13,163.08,151.31,143.56,140.39,137.55,130.03,129.74,127.30,126.33,124.05,123.12,122.76,120.70,119.02,112.07,108.98,101.97,61.14,21.38,10.96.

ESI-HRMS(C ₂₃H ₂₀N ₄O ₄S):m/z 447.11215(M-H ⁺).

The synthesis of embodiment 13, N-(4-(N-(3-((5-methyl isophthalic acid, 3,4-oxadiazole-2-base) methoxyl group)-9H-carbazole-1-base) amino-sulfonyl) phenyl) ethanamide (MI-13)

Intermediate compound I M9 (228mg, 0.7mmol) is dissolved in methyl alcohol (25mL), adds palladium carbon (10%, 44.4mg).Mixture is stirred overnight at room temperature in hydrogenation instrument (40psi).Filtering palladium carbon, is directly used in next step after filtrate is concentrated.

Reduzate is dissolved in pyridine (5mL), adds p-acetaminobenzenesulfonyl chloride (273.2mg, 1.17mmol) in batches.Mixture stirred overnight at room temperature.Steamed by solvent pyridine with surge pump, crude product silica gel column chromatography is separated (sherwood oil/acetone=3:2-1:1) and obtains white solid (182mg), and productive rate is 52.7%.

¹H NMR(400MHz,DMSO-d ₆):δ10.78(s,1H,carbazole NH),10.26(s,1H,N HCOCH ₃),9.97(s,1H,SO ₂NH),8.03(d,J＝7.84Hz,1H,ArH),7.73-7.67(m,5H,ArH),7.55(d,J＝8.20Hz,1H,ArH),7.38(td,1H,ArH),7.14(td,1H,ArH),6.75(d,J＝2.32Hz,1H,ArH),5.31(s,2H,CH ₂),2.53(s,3H,oxadiazoleCH ₃),2.04(s,3H,NHCOC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.46,165.15,163.04,151.21,143.71,140.45,133.28,129.75,128.58,126.46,124.26,122.69,122.10,120.76,119.12,118.94,112.08,109.54,102.75,61.21,24.56,10.95.

ESI-HRMS(C ₂₄H ₂₁N ₅O ₅S):m/z 492.13362(M+H ⁺)530.09093(M+K ⁺).

The synthesis of embodiment 14, N-(3-(N-(3-((5-methyl isophthalic acid, 3,4-oxadiazole-2-base) methoxyl group)-9H-carbazole-1-base) amino-sulfonyl) phenyl) ethanamide (MI-14)

Intermediate compound I M9 (149.6mg, 0.46mmol) is dissolved in methyl alcohol (20mL), adds palladium carbon (10%, 24.4mg).Mixture is stirred overnight at room temperature in hydrogenation instrument (40psi).Filtering palladium carbon, is directly used in next step after filtrate is concentrated.

Reduzate is dissolved in pyridine (5mL), adds 3-acetylsulphanilyl chloride (170mg, 0.73mmol) in batches.Mixture stirred overnight at room temperature.Steamed by solvent pyridine with surge pump, crude product silica gel column chromatography is separated (sherwood oil/acetone=3:2-1:1) and obtains white solid (122.5mg), and productive rate is 54.0%.

¹H NMR(400MHz,DMSO-d ₆):δ10.80(s,1H,carbazole N H),10.23(s,1H,N HCOCH ₃),10.18(s,1H,SO ₂N H),8.27(s,1H,ArH),8.04(d,J＝7.84Hz,1H,ArH),7.70-7.67(m,2H,ArH),7.56(d,J＝8.16Hz,1H,ArH),7.45(d,J＝4.80Hz,2H,ArH),7.39(t,1H,ArH),7.14(t,1H,ArH),6.74(d,J＝2.04Hz,1H,ArH),5.31(s,2H,CH ₂),2.53(s,3H,oxadiazoleCH ₃),2.04(s,3H,NHCOC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.29,165.17,163.02,151.20,140.47,140.36,140.32,130.10,129.88,126.50,124.30,123.28,122.66,121.82,121.63,120.78,119.13,117.36,112.09,109.74,102.87,61.15,24.47,10.96.

ESI-HRMS(C ₂₄H ₂₁N ₅O ₅S):m/z 490.11906(M-H ⁺).

Embodiment 15, N-(4-(N-(6-((5-methyl isophthalic acid; 3,4-oxadiazole-2-base) methoxyl group)-2,3; 4,9-tetrahydrochysene-1H-carbazole-8-base) amino-sulfonyl) phenyl) synthesis of ethanamide (MI-15)

The synthesis of the first step intermediate compound I M10

In the tetrahydrofuran solution (20mL) of IM2 (1.99g, 6.25mmol), drip the aqueous solution (20mL) of lithium hydroxide (570mg, 12.5mmol), dropwise rear stirring at room temperature 2 hours.React rear reaction solution 100mL diluted ethyl acetate, extracted three times (each 100mL) with saturated sodium bicarbonate solution.Merge aqueous phase, slowly regulate pH to 1 with the dilute hydrochloric acid of 10%, the yellow solid Büchner funnel suction filtration of precipitation, a small amount of water washing, then infrared lower oven dry.Be weighed as 1.527g, yield is 84.1%.

¹H NMR(400MHz,DMSO-d ₆):δ13.05(s,1H,COOH),11.39(s,1H,NH),7.53(d,J＝2.12Hz,1H,ArH),7.47(d,J＝2.04Hz,1H,ArH),4.79(s,2H,OC H ₂COOH),2.76(t,2H,1-C H ₂),2.62(t,2H,4-C H ₂),1.81(m,4H,2,3-C H ₂C H ₂).

The synthesis of second step intermediate compound I M11

By IM10 (1.1623g, 4mmol), acethydrazide (611.8mg, 8mmol), HOBt (822.3mg, 6mmol) and EDCI (1.15g, 3mmol) is dissolved in 16mL DMF.Mixture stirring at room temperature 7.5 hours, has reacted rear dropping 160mL water and product is separated out.Use Büchner funnel suction filtration, filter cake infrared drying after a small amount of water washing.Obtain red solid 1.316g, yield 94.9%.Be directly used in next step.

The synthesis of the 3rd step intermediate compound I M12

IM11 (703.2mg, 2mmol) is dissolved in phosphorus oxychloride (10mL), reacts and carry out under tube sealing condition, be heated to 100 degrees Celsius of reactions 4 hours.Be cooled to room temperature subsequently, concentrated solvent, add 2M sodium hydroxide solution and adjust pH for alkalescence, use acetic acid ethyl dissolution (150mL) subsequently.Organic phase first washes twice with 100mL 0.5M sodium hydroxide solution, uses anhydrous sodium sulfate drying subsequently, and after concentrated solvent, crude product silica gel column chromatography is separated (petrol ether/ethyl acetate=3:1) and obtains yellow solid, transforms completely.

¹H NMR(400MHz,DMSO-d ₆):δ11.44(s,1H,NH),7.67(d,J＝1.88Hz,1H,ArH),7.60(d,J＝1.64Hz,1H,ArH),5.46(s,2H,OC H ₂COOH),2.76(t,2H,1-C H ₂),2.62(t,2H,4-C H ₂),2.53(s,3H,CH ₃)1.81(m,4H,2,3-C H ₂C H ₂).

¹³C NMR(100MHz,DMSO-d ₆):δ165.24,162.91,150.04,140.54,132.63,131.46,125.17,112.92,110.34,105.19,61.32,23.41,22.98,22.92,20.72,10.95.

4th step

By intermediate compound I M12 (656.4mg; 2mmol) be dissolved in the mixed solution (1:1 of methyl alcohol and methylene dichloride; 50mL); with slowly dripping V-Brite B (2.165g half an hour under nitrogen protection condition; 12mmol) and the aqueous solution (7mL) of salt of wormwood (1.79g, 12mmol).Dropwise rear stirring at room temperature two hours, subsequently concentrated solvent, crude product 50mL water and 50mL methylene dichloride separatory, aqueous phase uses dichloromethane extraction twice again, each 50mL.Use anhydrous sodium sulfate drying after merging organic phase, after organic phase is concentrated, be directly used in next step.

Reduzate is dissolved in pyridine (5mL), slowly adds p-acetaminobenzenesulfonyl chloride (645mg, 2.76mmol).Mixture stirred overnight at room temperature.Steam except pyridine with surge pump, crude product is separated (sherwood oil/acetone=1:1-1:2) through silica gel column chromatography and obtains white solid 190mg, and yield is 19.2%.

¹H NMR(400MHz,DMSO-d ₆):δ10.29(s,1H,carbazole N H),10.19(s,1H,N HCOCH ₃),9.72(s,1H,SO ₂N H),7.69(t,4H,ArH),6.77(d,J＝1.88Hz,1H,ArH),6.49(d,J＝2.00Hz,1H,ArH),5.19(s,2H,OCH ₂),2.66(t,2H,1-C H ₂),2.52(m,5H,4-C H ₂&oxadiazoleC H ₃),2.05(s,3H,NHCOC H ₃),1.77(m,4H,2,3-C H ₂C H ₂).

¹³C NMR(100MHz,DMSO-d ₆):δ169.46,165.06,163.24,151.23,143.63,136.15,133.41,128.94,128.51,124.77,121.93,118.92,109.44,103.56,99.04,60.97,24.57,23.28,23.21,23.16,21.06,10.93.

ESI-HRMS(C ₂₄H ₂₅N ₅O ₅S):m/z 494.15036(M-H ⁺).

The synthesis of embodiment 16, ethyl 2-((1-(4-Methyl benzenesulfonyl amido)-9H-carbazole-3-base) sulfo-) acetic ester (MI-16)

The synthesis of the first step intermediate compound I M13

By to the concentrated hydrochloric acid suspension (50mL) of amino m-nitrophenol (4.6192g, 30mmol) in stirring at room temperature half hour until black suspendible solid gradually becomes white.Be cooled to-10 degrees Celsius subsequently, the aqueous solution (3.5mL) slowly dripping Sodium Nitrite (2.4897g, 36mmol) in suspension carries out diazotization reaction.The concentrated hydrochloric acid solution (10mL) of tin protochloride (13.6g, 60mmol) is slowly dripped after 1 hour.Dropwise mixture and stir half an hour at-10 degrees Celsius, rise to room temperature subsequently and continue stirring 3 hours.

With 50mL alcohol dilution said mixture, then slowly drip pimelinketone (3.4mL, 33mmol), be heated to 90 degrees Celsius of backflows and spend the night.Be cooled to room temperature, a small amount of water washing of black solid of precipitation, infrared drying subsequently, is weighed as 2.34g, and yield is 38.1%.

¹H NMR(400MHz,DMSO-d ₆):δ11.19(s,1H,NH),9.44(s,1H,OH),7.43(s,1H,ArH),7.22(s,1H,ArH),2.74(t,2H,1-CH ₂),2.58(t,2H,4-CH ₂),1.80(m,4H,2,3-CH ₂).

¹³C NMR(100MHz,DMSO-d ₆):δ149.91,139.70,132.79,131.49,124.07,112.58,109.62,104.86,23.40,23.07,23.00,20.69.

The synthesis of second step intermediate compound I M14

Intermediate compound I M13 (116.6mg, 0.5mmol) is suspended in 5mL methylene dichloride, under condition of ice bath, drips 2,6-lutidine (192uL, 1.65mmol), trifluoromethanesulfanhydride anhydride (138uL, 1.1mmol) is slowly dripped with 5 minutes subsequently.Mixed solution room temperature for overnight, uses 20mL dchloromethane subsequently, and organic phase 20mL 2N dilute hydrochloric acid washes three times, and 20mL saturated common salt is washed once, anhydrous sodium sulfate drying.Concentrated solvent obtains yellow solid (transforming completely).

¹H NMR(400MHz,DMSO-d ₆):δ11.93(s,1H,NH),7.98(s,2H,ArH),2.80(t,2H,1-CH ₂),2.66(t,2H,4-CH ₂),1.82(m,4H,2,3-CH ₂).

¹³C NMR(100MHz,DMSO-d ₆):δ142.41,140.63,132.47,131.23,127.63,118.78(q,J＝320Hz),118.10,111.63,110.00,23.42,22.77,22.71,20.55.

The synthesis of the 3rd step intermediate compound I M15

By previous step gained intermediate compound I M14 (0.5mmol) and DDQ heated overnight at reflux in 5mL toluene, cool to room temperature subsequently.Reaction solution 50mL diluted ethyl acetate, washes 5 times with 50mL saturated sodium bicarbonate solution until aqueous phase is colourless, organic phase anhydrous sodium sulfate drying.Concentrated solvent, gained crude product is separated (methylene dichloride/sherwood oil=1:3) through silica gel column chromatography, and obtain yellow solid 126.5mg, yield is 77.5%.

¹H NMR(400MHz,DMSO-d ₆):δ12.46(s,1H,NH),8.89(s,1H,ArH),8.37(s,2H,ArH),7.78(d,1H,ArH),7.59(t,1H,ArH),7.35(t,1H,ArH).

¹³C NMR(100MHz,DMSO-d ₆):δ142.16,140.37,132.47,131.24,129.04,128.47,121.93,121.65,121.57,121.45,118.81(q,J＝320Hz),114.87,113.47.

19F NMR(376MHz,DMSO-d6):δ-72.41.

The synthesis of the 4th step intermediate compound I M16

By Pd2 (dba) 3 (2311.4mg, 0.25mmol) and Xantphos (292.4mg, 0.5mmol) pre-mixing 1 hour in anhydrous dioxane under the condition of argon shield.Add intermediate compound I M15 (360mg, 1mmol) subsequently to reaction tubes, again vacuumize rear perfusion argon gas, in triplicate.Then add DIPEA (0.35mL) and ethyl thioglycolate (0.11mL) with syringe, be heated to 100 degrees Celsius of reactions 20 hours.Reaction terminates rear 50mL diluted ethyl acetate reaction solution, washs three times, anhydrous sodium sulfate drying after filtration catalizer with 50mL 2N dilute hydrochloric acid.Be separated (ethyl acetate/petroleum ether=1:20-1:10) with silica gel column chromatography after concentrated solvent, obtain yellow solid 250.0mg, yield is 75.6%.

¹H NMR(400MHz,DMSO-d ₆):δ12.25(s,1H,carbazoleNH),8.76(s,1H,ArH),8.35(s,1H,ArH),8.27(d,J＝7.76Hz,1H,ArH),7.75(d,J＝8.12Hz,1H,ArH),7.54(t,1H,ArH),7.32(t,1H,ArH),4.07(q,J＝7.12Hz,2H,C H ₂CH ₃),3.93(s,2H,SC H ₂),1.10(t,J＝7.08Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.75,141.58,132.83,131.95,131.59,128.42,128.30,124.83,123.72,121.48,121.42,113.22,61.35,37.72,14.38.

5th step

By intermediate compound I M16 (250mg; 0.75mmol) be dissolved in the mixed solution (1:1 of methyl alcohol and methylene dichloride; 50mL); with slowly dripping V-Brite B (816.4mg half an hour under nitrogen protection condition; 4.64mmol) and the aqueous solution (7mL) of salt of wormwood (650.6mg, 4.67mmol).Dropwise rear stirring at room temperature two hours, subsequently concentrated solvent, crude product 50mL water and 50mL methylene dichloride separatory, aqueous phase uses dichloromethane extraction twice again, each 50mL.Use anhydrous sodium sulfate drying after merging organic phase, after organic phase is concentrated, be directly used in next step.

Reduzate is dissolved in pyridine (5mL), slowly adds p-methyl benzene sulfonic chloride (239.1mg, 1.25mmol).Mixture stirred overnight at room temperature.Steam except pyridine with surge pump, crude product is separated (sherwood oil/acetone=3:1) through silica gel column chromatography and obtains white solid 219.8mg, and yield is 63.9%.

¹H NMR(400MHz,DMSO-d ₆):δ11.10(s,1H,carbazoleNH),9.94(s,1H,SO ₂NH),8.09(d,J＝8.20Hz,1H,ArH),8.07(s,1H,ArH),7.64(d,J＝7.96Hz,2H,ArH),7.59(d,J＝8.12Hz,1H,ArH),7.42(t,1H,ArH),7.33(d,J＝7.84Hz,2H,ArH),7.18(t,1H,ArH),7.00(s,1H,ArH),4.03(q,J＝7.08Hz,2H,C H ₂CH ₃),3.58(s,2H,SCH ₂),2.33(s,3H,PhC H ₃),1.09(t,J＝7.12Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.69,143.84,140.31,136.85,134.70,130.03,127.45,126.81,124.97,124.29,123.03,122.52,122.18,121.76,120.87,119.78,112.17,61.15,38.56,21.40,14.40.

ESI-HRMS(C ₂₃H ₂₂N ₂O ₄S ₂):m/z 453.09482(M-H ⁺).

The synthesis of embodiment 17, ethyl 2-((1-(4-P-acetamido benzene sulfonyl amido)-9H-carbazole-3-base) sulfo-) acetic ester (MI-17)

Reduzate is dissolved in pyridine (5mL), slowly adds p-acetaminobenzenesulfonyl chloride (301.2mg, 1.275mmol).Mixture stirred overnight at room temperature.Steam except pyridine with surge pump, crude product is separated (sherwood oil/acetone=2:1-3:2) through silica gel column chromatography and obtains white solid 186mg, and yield is 49.4%.

¹H NMR(400MHz,DMSO-d ₆):δ11.09(s,1H,carbazoleNH),10.26(s,1H,N HCOCH ₃),9.89(s,1H,SO ₂NH),8.09(d,J＝8.08Hz,1H,ArH),8.07(s,1H,ArH),7.69(t,4H,ArH),7.59(d,J＝8.08Hz,1H,ArH),7.42(t,1H,ArH),7.18(t,1H,ArH),7.03(s,1H,ArH),4.02(q,J＝7.04Hz,2H,C H ₂CH ₃),3.59(s,2H,SCH ₂),2.05(s,3H,NHCOC H ₃),1.07(t,J＝7.04Hz,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.72,169.43,143.73,140.30,134.75,133.16,128.63,126.80,124.96,124.43,122.98,122.57,122.19,121.80,120.86,119.77,118.86,112.16,61.13,38.58,24.57,14.39.

ESI-HRMS(C ₂₄H ₂₃N ₃O ₅S ₂):m/z 496.10064(M-H ⁺).

The synthesis of embodiment 18,2-((1-(4-Methyl benzenesulfonyl amido)-9H-carbazole-3-base) sulfo-) acetic acid (MI-18)

In the tetrahydrofuran solution (1mL) of MI-16 (89.5mg, 0.2mmol), drip the aqueous solution (1mL) of 1N sodium hydroxide, dropwise rear stirring at room temperature 0.5 hour.React rear reaction solution 20mL diluted ethyl acetate, extracted three times (each 20mL) with saturated sodium bicarbonate solution.Merge aqueous phase, slowly regulate pH to 1 with the dilute hydrochloric acid of 10%, the white solid Büchner funnel suction filtration of precipitation, a small amount of water washing, then infrared lower oven dry.Be weighed as 74.2mg, yield is 88.4%.

¹H NMR(400MHz,DMSO-d ₆):δ12.51(s,1H,COOH),11.10(s,1H,carbazoleNH),10.03(s,1H,SO ₂NH),8.08(d,J＝7.96Hz,1H,ArH),8.05(s,1H,ArH),7.65(d,J＝7.96Hz,2H,ArH),7.58(d,J＝7.96Hz,1H,ArH),7.41(t,1H,ArH),7.32(d,J＝7.96Hz,2H,ArH),7.18(t,1H,ArH),7.01(s,1H,ArH),3.55(s,2H,SCH ₂),2.32(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ171.06,143.85,140.29,136.82,134.46,130.04,127.47,126.76,124.96,123.80,123.75,122.18,121.79,121.72,120.87,119.70,112.14,38.85,21.42.

ESI-HRMS(C ₂₁H ₁₈N ₂O ₄S ₂):m/z 425.06352(M-H ⁺).

The synthesis of embodiment 19,2-((1-(4-P-acetamido benzene sulfonyl amido)-9H-carbazole-3-base) sulfo-) acetic acid (MI-19)

In the tetrahydrofuran solution (1mL) of MI-17 (84.7mg, 0.15mmol), drip the aqueous solution (1mL) of 1N sodium hydroxide, dropwise rear stirring at room temperature 0.5 hour.React rear reaction solution 20mL diluted ethyl acetate, extracted three times (each 20mL) with saturated sodium bicarbonate solution.Merge aqueous phase, slowly regulate pH to 1 with the dilute hydrochloric acid of 10%, the white solid Büchner funnel suction filtration of precipitation, a small amount of water washing, then infrared lower oven dry.Be weighed as 65.8mg, yield is 82.3%.

¹H NMR(400MHz,DMSO-d ₆):δ12.23(br s,1H,COOH),11.09(s,1H,carbazoleNH),10.26(s,1H,N HCOCH ₃),9.97(br s,1H,SO ₂NH),8.08(d,J＝8.00Hz,1H,ArH),8.05(s,1H,ArH),7.69(s,4H,ArH),7.58(d,J＝7.92Hz,1H,ArH),7.41(t,1H,ArH),7.17(t,1H,ArH),7.03(s,1H,ArH),3.56(s,2H,ArH),2.05(s,3H,ArH).

¹³C NMR(100MHz,DMSO-d ₆):δ171.11,169.45,143.70,140.28,134.47,133.20,128.66,126.74,124.95,123.84,123.80,122.18,121.83,121.66,120.87,119.69,118.90,112.13,38.92,24.57.

ESI-HRMS(C ₂₂H ₁₉N ₃O ₅S ₂):m/z 468.06934(M-H ⁺).

The synthesis of embodiment 20, ethyl 2-((1-(4-Methyl benzenesulfonyl amido)-9H-carbazole-3-base) alkylsulfonyl) acetic ester (MI-20)

MI-16 (100.3mg, 0.22mmol) is dissolved in 4mL acetone, drips the aqueous solution of 4mL oxone (700.4mg, 1.1mmol) subsequently.Mixed solution stirred overnight at room temperature, subsequently with the dilution of 4mL water, filters to obtain white solid, washing, dries.Solid silica gel column chromatography purification (sherwood oil/acetone=3:1) obtains white solid 52.7mg, and yield is 49.3%.

¹H NMR(400MHz,DMSO-d ₆):δ11.66(s,1H,carbazole NH),10.17(s,1H,SO ₂NH),8.55(d,J＝1.52Hz,1H,ArH),8.26(d,J＝7.84Hz,1H,ArH),7.68(d,J＝8.20Hz,1H,ArH),7.64(d,J＝8.28Hz,2H,ArH),7.52(td,1H,ArH),7.37(d,J＝1.64Hz,1H,ArH),7.34(d,J＝8.00Hz,2H,ArH),7.29(td,1H,ArH),4.35(s,2H,SO ₂CH ₂),3.98(q,J＝7.12Hz,2H,C H ₂CH ₃),2.32(s,3H,PhC H ₃),0.97(t,J＝7.12Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ162.96,144.09,140.79,137.98,136.45,130.10,129.07,127.73,127.52,124.15,122.53,121.60,121.37,120.81,119.85,119.39,112.67,61.85,61.32,21.40,14.09.

ESI-HRMS(C ₂₃H ₂₂N ₂O ₆S ₂):m/z 485.08465(M-H ⁺).

The synthesis of embodiment 21,2-((1-(4-Methyl benzenesulfonyl amido)-9H-carbazole-3-base) alkylsulfonyl) acetic acid (MI-21)

In the tetrahydrofuran solution (1mL) of MI-20 (114.5mg, 0.235mmol), drip the aqueous solution (1mL) of 1N sodium hydroxide, dropwise rear stirring at room temperature 0.5 hour.React rear reaction solution 20mL diluted ethyl acetate, extracted three times (each 20mL) with saturated sodium bicarbonate solution.Merge aqueous phase, slowly regulate pH to 1 with the dilute hydrochloric acid of 10%, the white solid Büchner funnel suction filtration of precipitation, a small amount of water washing, then infrared lower oven dry.Be weighed as 94.6mg, yield is 87.7%.

¹H NMR(400MHz,DMSO-d ₆):δ11.78(s,1H,carbazole NH),10.37(s,1H,SO ₂NH),8.54(s,1H,ArH),8.22(d,J＝7.80Hz,1H,ArH),7.65-7.63(m,3H,ArH),7.48(t,1H,ArH),7.43(d,J＝1.64Hz,1H,ArH),7.30-7.24(m,3H,ArH),4.19(s,2H,SO ₂CH ₂),2.29(s,3H,PhC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ164.58,143.97,140.78,137.81,136.56,130.04,129.82,127.58,127.52,123.99,122.56,121.66,121.29,120.64,119.57,119.40,112.60,62.00,21.39.

ESI-HRMS(C ₂₁H ₁₈N ₂O ₆S ₂):m/z 457.05335(M-H ⁺).

The synthesis of embodiment 22, ethyl 2-((1-(4-P-acetamido benzene sulfonyl amido)-9H-carbazole-3-base) alkylsulfonyl) acetic ester (MI-22)

MI-17 (90mg, 0.18mmol) is dissolved in 4mL acetone, drips the aqueous solution of 4mL oxone (558.8mg, 0.9mmol) subsequently.Mixed solution stirred overnight at room temperature, subsequently with the dilution of 4mL water, filters to obtain white solid, washing, dries.Solid silica gel column chromatography purification (methylene dichloride/acetone=5:1-3:1) obtains white solid 43.3mg, and yield is 45.2%.

¹H NMR(400MHz,DMSO-d ₆):δ11.67(s,1H,carbazole NH),10.30(s,1H,N HCOCH ₃),10.14(s,1H,SO ₂NH),8.56(s,1H,ArH),8.27(d,J＝7.80Hz,1H,ArH),7.72-7.67(m,5H,ArH),7.52(t,J＝7.72Hz,ArH),7.38(s,1H,ArH),7.28(t,J＝7.36Hz,ArH),4.38(s,2H,SO ₂C H ₂),3.97(q,J＝7.12Hz,2H,C H ₂CH ₃),2.05(s,3H,NHCOC H ₃),0.96(t,J＝7.08Hz,3H,CH ₂C H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.47,162.97,143.88,140.79,138.07,132.66,129.01,128.74,127.72,124.13,122.52,121.61,121.37,120.79,119.89,119.56,118.88,112.67,61.84,61.34,24.57,14.07.

ESI-HRMS(C ₂₄H ₂₃N ₃O ₇S ₂):m/z 528.09047(M-H ⁺).

Embodiment 23, N-(4-(N-(3-(((3-methyl isophthalic acid; 2,4-oxadiazole-5-base) methyl) sulfo-)-9H-carbazole-1-base) amino-sulfonyl) phenyl) synthesis of ethanamide (MI-23)

The synthesis of the first step intermediate compound I M18

In the tetrahydrofuran solution (6mL) of IM17 (645mg, 2mmol), drip the aqueous solution (6mL) of lithium hydroxide (180.2mg, 4mmol), dropwise rear stirring at room temperature 1 hour.React rear reaction solution 50mL diluted ethyl acetate, extracted three times (each 40mL) with saturated sodium bicarbonate solution.Merge aqueous phase, slowly regulate pH to 1 with the dilute hydrochloric acid of 10%, the yellow solid Büchner funnel suction filtration of precipitation, a small amount of water washing, then infrared lower oven dry.Be weighed as 580.4mg, yield is 98.3%.

¹H NMR(400MHz,DMSO-d ₆):δ12.81(br s,1H,COOH),12.21(s,1H,NH),8.73(d,J＝1.32Hz,1H,ArH),8.33(d,J＝1.64Hz,1H,ArH),8.25(d,J＝7.80Hz,1H,ArH),7.74(d,J＝8.12Hz,1H,ArH),7.53(td,1H,ArH),7.30(td,1H,ArH),3.88(s,2H,CH ₂).

¹³C NMR(100MHz,DMSO-d ₆):δ171.10,141.56,132.65,131.97,130.91,128.39,128.24,124.50,124.16,121.46,121.41,121.33,113.18,38.03.

The synthesis of second step intermediate compound I M19

By intermediate compound I M18 (530.2mg, 1.75mmol), HOBt (367.4mg, 2.63mmol) and EDCI (527.8mg, 2.63mmol) be stirring at room temperature half an hour in anhydrous acetonitrile (6mL) first.Drip the acetonitrile solution (4mL) of acetyl amidoxime (259.3mg, 3.5mmol) subsequently, then mixture flows through night next time in heating 90 degrees celsius.Concentrated solvent, solid silica gel column chromatography is separated (petrol ether/ethyl acetate=3:1) and obtains orange solids 455.1mg, and yield is 76.2%.

¹H NMR(400MHz,DMSO-d ₆):δ12.28(s,1H,carbazole NH),8.74(s,1H,ArH),8.30(s,1H,ArH),8.25(d,J＝7.64Hz,1H,ArH),7.75(d,J＝8.00Hz,1H,ArH),7.54(t,J＝7.24Hz,1H,ArH),7.32(t,J＝7.32Hz,1H,ArH),4.60(s,2H,SCH ₂),2.25(s,3H,oxadiazoleCH ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ176.95,167.54,141.55,133.05,132.27,131.93,128.48,128.36,125.65,122.24,121.50,121.47,121.42,113.25,30.01,11.50.

3rd step

By intermediate compound I M19 (340.36mg; 1mmol) be dissolved in the mixed solution (1:1 of methyl alcohol and methylene dichloride; 50mL); with slowly dripping V-Brite B (1.08g half an hour under nitrogen protection condition; 6.18mmol) and the aqueous solution (8mL) of salt of wormwood (868mg, 6.23mmol).Dropwise rear stirring at room temperature two hours, subsequently concentrated solvent, crude product 50mL water and 50mL methylene dichloride separatory, aqueous phase uses dichloromethane extraction twice again, each 50mL.Use anhydrous sodium sulfate drying after merging organic phase, after organic phase is concentrated, be directly used in next step.

Reduzate is dissolved in pyridine (5mL), slowly adds p-acetaminobenzenesulfonyl chloride (397mg, 1.7mmol).Mixture stirred overnight at room temperature.Steam except pyridine with surge pump, crude product is separated (sherwood oil/acetone=3:2-1:1) through silica gel column chromatography and obtains white solid 174.4mg, and yield is 34.4%.

¹H NMR(400MHz,DMSO-d ₆):δ11.14(s,1H,carbazole NH),10.25(s,1H,N HCOCH ₃),9.89(s,1H,SO ₂NH),8.08(d,J＝7.88Hz,1H,ArH),8.07(d,J＝1.44Hz,1H,ArH),7.70-7.65(m,4H,ArH),7.59(d,J＝8.16Hz,1H,ArH),7.42(td,1H,ArH),7.19(td,1H,ArH),7.02(d,J＝1.60Hz,1H,ArH),4.27(s,2H,SCH ₂),2.27(s,3H,oxadiazoleCH ₃),2.04(s,3H,NHCOC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ177.03,169.43,167.47,143.73,140.30,135.08,133.08,128.64,126.91,125.02,123.32,122.17,121.87,121.76,120.92,119.87,118.85,112.19,30.94,24.56,11.55.

ESI-HRMS(C ₂₄H ₂₁N ₅O ₄S ₂):m/z 506.09622(M-H ⁺).

Embodiment 24, N-(4-(N-(3-(((3-methyl isophthalic acid; 2,4-oxadiazole-5-base) methyl) alkylsulfonyl)-9H-carbazole-1-base) amino-sulfonyl) phenyl) synthesis of ethanamide (MI-24)

MI-23 (82.8mg, 0.16mmol) is dissolved in 4mL acetone, drips the aqueous solution of 4mL oxone (985.6mg, 1.6mmol) subsequently.Mixed solution stirred overnight at room temperature, subsequently with the dilution of 8mL water, filters to obtain white solid, washing, dries.Solid silica gel column chromatography purification (sherwood oil/acetone=3:1-1:1) obtains white solid 43.3mg, and yield is 50.2%.

¹H NMR(400MHz,DMSO-d ₆):δ11.67(s,1H,carbazole NH),10.28(s,1H,N HCOCH ₃),10.11(s,1H,SO ₂NH),8.53(s,1H,ArH),8.26(d,J＝7.72Hz,1H,ArH),7.70-7.65(m,5H,ArH),7.53(t,1H,ArH),7.33(s,1H,ArH),7.29(t,1H,ArH),5.15(s,2H,SO ₂C H ₂),2.29(s,3H,oxadiazoleCH ₃),2.03(s,3H,NHCOC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.62,169.47,167.94,143.90,140.80,138.23,132.68,128.70,128.14,127.84,124.30,122.50,121.72,121.41,120.90,119.94,119.52,118.87,112.70,53.63,24.56,11.50.

ESI-HRMS(C ₂₄H ₂₁N ₅O ₆S ₂):m/z 538.08605(M-H ⁺).

Embodiment 25, N-(4-(N-(3-(((5-methyl isophthalic acid; 3,4-oxadiazole-2-base) methyl) sulfo-)-9H-carbazole-1-base) amino-sulfonyl) phenyl) synthesis of ethanamide (MI-25)

The synthesis of the first step intermediate compound I M20

IM18 (305.4mg, 1mmol) and acethydrazide (125.9mg, 1.5mmol) are dissolved in phosphorus oxychloride (8mL).Mixed solution reacts and spends the night under 100 degrees celsius.Concentrated solvent, crude product is separated through silica gel column chromatography and obtains yellow solid 168.2mg, and yield is 48.9%.

¹H NMR(400MHz,DMSO-d ₆):δ12.29(s,1H,carbazole NH),8.74(s,1H,ArH),8.30(s,1H,ArH),8.26(d,J＝7.80Hz,1H,ArH),7.75(d,J＝7.80Hz,1H,ArH),7.54(t,J＝7.80Hz,1H,ArH),7.32(t,J＝7.80Hz,1H,ArH),4.53(s,2H,SCH ₂),2.44(s,3H,oxadiazoleCH ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ164.70,164.13,141.54,133.05,132.54,131.92,128.47,128.35,125.82,122.21,121.49,121.45,113.25,29.32,10.88.

Second step

By intermediate compound I M20 (173.4mg; 0.5mmol) be dissolved in the mixed solution (1:1 of methyl alcohol and methylene dichloride; 28mL); with slowly dripping V-Brite B (544mg half an hour under nitrogen protection condition; 3.1mmol) and the aqueous solution (3mL) of salt of wormwood (432.3mg, 3.1mmol).Dropwise rear stirring at room temperature two hours, subsequently concentrated solvent, crude product 30mL water and 30mL methylene dichloride separatory, aqueous phase uses dichloromethane extraction twice again, each 30mL.Use anhydrous sodium sulfate drying after merging organic phase, after organic phase is concentrated, be directly used in next step.

Reduzate is dissolved in pyridine (5mL), slowly adds p-acetaminobenzenesulfonyl chloride (206.4mg, 0.85mmol).Mixture stirred overnight at room temperature.Steam except pyridine with surge pump, crude product is separated (sherwood oil/acetone=3:2-1:1) through silica gel column chromatography and obtains white solid 94.6mg, and yield is 36.6%.

¹H NMR(400MHz,DMSO-d ₆):δ11.14(s,1H,carbazole NH),10.27(s,1H,N HCOCH ₃),9.89(s,1H,SO ₂NH),8.08(d,J＝7.80Hz,1H,ArH),8.04(d,J＝1.00Hz,1H,ArH),7.67(q,J＝6.64Hz,4H,ArH),7.58(d,J＝8.12Hz,1H,ArH),7.42(t,J＝7.48Hz,1H,ArH),7.18(t,J＝7.40Hz,1H,ArH),6.99(d,J＝1.32Hz,1H,ArH),4.19(s,2H,SCH ₂),2.43(s,3H,oxadiazoleCH ₃),2.03(s,3H,NHCOC H ₃).

¹³C NMR(100MHz,DMSO-d ₆):δ169.44,164.55,164.21,143.70,140.28,135.10,133.06,128.68,126.88,125.40,125.01,123.69,122.18,121.81,121.58,120.93,119.85,118.85,112.18,30.27,24.56,10.88.

ESI-HRMS(C ₂₄H ₂₁N ₅O ₄S ₂):m/z 506.09622(M-H ⁺).

Embodiment 26, compound are to the restraining effect of disactivation MEK1

HTRF (homogeneous time-resolved fluorescence assay) combine with technique FRET (fluorescence resonance energy transfer) (FRET) and time resolved fluorescence (TRF) two kinds of technology are adopted in inhibit activities test experiments.This technology make use of the chela of the Eu element with cave-shaped structure and marker as a donor (Donor), is based on the FRET (fluorescence resonance energy transfer) (FRET) between the donor of Eu kryptofix 222 and acceptor (the second fluorescent marker).In FRET (fluorescence resonance energy transfer) experiment, the life-span of acceptor emission fluorescence is equal to the life-span of the emitting fluorescence of donor.Because the fluorescence decay cycle of Eu is longer, so know from experience inducing receptor emitting fluorescence for a long time containing the confession of Eu, acceptor excites the fluorescence of rear generation just can continue the long period, just can be distinguished the fluorescence of those self scatterings short-life like this by temporal resolution, just be easy to like this distinguish FRET signal from short life fluorescence background.

When cause due to bio-molecular interaction two fluorophors close to time, discharged by the portion of energy that kryptofix 222 is caught when exciting, emission wavelength is 615nm; Another part energy trasfer is on acceptor (acceptor), and emission wavelength is 665nm.The FRET that the utilizing emitted light of 665nm is only caused by donor (donor) produces.So, when bio-molecular interaction, there are two exciting light 615nm and 665nm; When there is not interaction, only has 615nm exciting light.Experimental principle as shown in Figure 1.

If compound has restraining effect to npMEK1, then can suppress the phosphorylation of npMEK1, the anti-phosphorus antibody be combined with MEK1 accordingly reduces, and the signal of the MAb Anti GST-XL665 therefore excited also can correspondingly reduce.By detecting the ratio of the signal of 668nm and the signal of 620nm, assessing compound is to the restraining effect of npMEK1.

Concrete operation step is divided into two steps, kinase reaction step and detection reaction step respectively, kinase reaction step is the step of BRAF phosphorylation MEK1, add by kinase reaction damping fluid 50mM pH=7.0HEPESbuffer, 10mM MgCl2,1mM DTT, 0.5mM Orthovanadate, the compound of 0.01%BSA dilution, 0.48ng/uL active BRAF (09-122) (Carna), 40nM inactive MEK1GSTtagged-(07-141-10) (Carna), 100 μMs of ATP, room temperature reaction 2 hours; Second step is detection reaction step, add cisbio company antibody MAb Anti-phospho MEK1/2-K (61P17KAE) and MAb AntiGST-XL665 (61GSTXLA), react after 3 hours and adopt microplate reader FlexStation 3 (Molecular Devices) detection signal, inhibiting rate method of calculation are as follows:

Mapping software GraphPad Prism 6 pairs of data are used to process.Experiment repetition 3 times.Part of compounds Activity Results is as shown in the table.

Table 1, the compounds of this invention are to the inhibit activities of disactivation MEK1

The foregoing is only the preferred embodiments of the present invention, is only illustrative for the purpose of the present invention, and nonrestrictive; Those of ordinary skill in the art understand, and can carry out many changes in the spirit and scope that the claims in the present invention limit to it, amendment, and even equivalence is changed, but all will fall within the scope of protection of the present invention.

Claims

1. there is the compound of MEK1 inhibit activities, and pharmacy acceptable salt or prodrug, it is characterized in that the structure of described compound is such as formula shown in I:

Wherein, A ring is phenyl ring or tetrahydrobenzene;

X is oxygen, sulphur, carbon, NH, SO, SO ₂;

N is 1,2,3 or 4;

R ₆, R ₇be selected from hydrogen separately respectively, C _1-3alkyl.

2. compound as claimed in claim 1, and pharmacy acceptable salt or prodrug, is characterized in that described C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group comprise methyl, ethyl, propyl group, sec.-propyl, normal-butyl, 2-methyl-propyl, 2-butyl, tertiary butyl, n-pentyl, methoxy methylol, cyclopropyl, Cvclopropvlmethvl, adamantyl and the monosubstituted or polysubstituted above-mentioned group through halogen, hydroxyl, amino, cyano group, nitro, carboxyl.

3. compound as claimed in claim 1, and pharmacy acceptable salt or prodrug, is characterized in that described C ₂-C ₇thiazolinyl be aliphatic alkyl containing the straight or branched containing 2-7 carbon atom in C=C key simultaneously chain, comprise vinyl, propenyl, butenyl, isobutenyl, 3-methyl-2-butene base, just pentenyl, heptenyl, cyclopropylethenyl, cyclohexyl propenyl and by single or multiple halogen, hydroxyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group; Described alkynyl refers to the aliphatic alkyl containing the straight or branched containing 2-7 carbon atom in carbon-to-carbon triple bond simultaneously chain, comprises ethynyl, proyl, positive butynyl, butynyl, 3-methyl-2-butyne base, just pentynyl etc. and through single or multiple halogen, hydroxyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

4. compound as claimed in claim 1, and pharmacy acceptable salt or prodrug, it is characterized in that described substituted or unsubstituted aryl refers to 6-14 unit monocycle or bicyclic aromatic group, comprise phenyl, naphthyl and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, kharophen, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group, wherein C ₁-C ₄alkyl comprise methyl, ethyl.

5. compound as claimed in claim 1, and pharmacy acceptable salt or prodrug, it is characterized in that described substituted or unsubstituted heteroaryl refers to independently be selected from containing 1-5 heteroatomic 4-10 unit's monocycle or the bicyclic aromatic group of N, O and S, comprise pyrryl, pyridyl, imidazolyl, indyl and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

6. compound as claimed in claim 1, and pharmacy acceptable salt or prodrug, it is characterized in that described substituted or unsubstituted heterocyclic radical refers to that preferred heteroatoms comprises N, O, S containing one or more heteroatomic compound in its ring structure.

7. compound as claimed in claim 6, and pharmacy acceptable salt or prodrug, it is characterized in that described substituted or unsubstituted heterocyclic radical is saturated or the aromatics of fractional saturation, preferably five yuan and single six-membered rings heterocyclic radical and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

8. compound as claimed in claim 1, and pharmacy acceptable salt or prodrug, is characterized in that described halogen comprises fluorine, chlorine, bromine and iodine.

9. compound as claimed in claim 1, and pharmacy acceptable salt or prodrug, is characterized in that described replacement or unsubstituted cycloalkyl refer to C ₃-C ₁₀saturated monocycle or many rings ring system, comprise cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl, adamantyl and through single or multiple halogen, hydroxyl, C ₁-C ₄alkyl, C ₁-C ₄alkoxyl group, amino, cyano group, nitro, carboxyl, aldehyde radical replace above-mentioned group.

10. prepare compound according to claim 1, and the method for pharmacy acceptable salt or prodrug, it is characterized in that in described formula I structure, A ring is phenyl ring, R ₃for the preparation method of the compound of hydroxyl, comprise the steps:

Wherein, R is hydrogen atom, C ₁-C ₁₀straight chain, branched-chain alkyl, substituted alkyl or cycloalkyl group, C ₂-C ₇alkenyl or alkynyl, replace or unsubstituted cycloalkyl, tertiary butyloxycarbonyl acyl group, benzyloxy carbonyl acyl group, benzyl, substituted benzyl, the substituted or unsubstituted aryl in optional position, heterocyclic radical, heteroaryl;

X is oxygen, sulphur, carbon, NH, SO, SO ₂;

N is 1,2,3 or 4;

11. preparation compound according to claim 1, and the method for pharmacy acceptable salt or prodrug, is characterized in that R in described formula I structure ₁, R ₃for hydrogen atom, X is the preparation method of the compound of Sauerstoffatom, comprises the steps:

Wherein, A ring is phenyl ring or tetrahydrobenzene;

N is 1,2,3 or 4;

12. preparation compound according to claim 1, and the method for pharmacy acceptable salt or prodrug, is characterized in that R in described formula I structure ₁, R ₃for hydrogen atom, X is the preparation method of the compound of sulphur atom or sulfone, comprises the steps:

Wherein, A ring is phenyl ring or tetrahydrobenzene;

X is S or SO ₂;

N is 1,2,3 or 4;

Compound described in 13. any one of claim 1-9, and pharmacy acceptable salt or prodrug are preparing the application in mek inhibitor.