CN106905347A - BRD4 inhibitor and its application in anti-tumor medicine - Google Patents
BRD4 inhibitor and its application in anti-tumor medicine Download PDFInfo
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- CN106905347A CN106905347A CN201710253407.9A CN201710253407A CN106905347A CN 106905347 A CN106905347 A CN 106905347A CN 201710253407 A CN201710253407 A CN 201710253407A CN 106905347 A CN106905347 A CN 106905347A
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- 0 Cc1cccc(*)c1 Chemical compound Cc1cccc(*)c1 0.000 description 4
- IWDCLRJOBJJRNH-UHFFFAOYSA-N Cc(cc1)ccc1O Chemical compound Cc(cc1)ccc1O IWDCLRJOBJJRNH-UHFFFAOYSA-N 0.000 description 2
- BPRYUXCVCCNUFE-UHFFFAOYSA-N Cc(cc1C)cc(C)c1O Chemical compound Cc(cc1C)cc(C)c1O BPRYUXCVCCNUFE-UHFFFAOYSA-N 0.000 description 2
- NZVZVGPYTICZBZ-UHFFFAOYSA-N C(c1ccccc1)N1CCCCC1 Chemical compound C(c1ccccc1)N1CCCCC1 NZVZVGPYTICZBZ-UHFFFAOYSA-N 0.000 description 1
- UVTJGLSUPAHUTC-SNCPXTGUSA-N CC/C(/F)=C\C(\F)=C/C Chemical compound CC/C(/F)=C\C(\F)=C/C UVTJGLSUPAHUTC-SNCPXTGUSA-N 0.000 description 1
- RKZMLBSCCJJOGT-UHFFFAOYSA-N CC1C=CC(Br)=CC1 Chemical compound CC1C=CC(Br)=CC1 RKZMLBSCCJJOGT-UHFFFAOYSA-N 0.000 description 1
- ZZLCFHIKESPLTH-UHFFFAOYSA-N Cc(cc1)ccc1-c1ccccc1 Chemical compound Cc(cc1)ccc1-c1ccccc1 ZZLCFHIKESPLTH-UHFFFAOYSA-N 0.000 description 1
- UHIGHLGTNVYXOP-UHFFFAOYSA-N Cc(cc1F)cc(F)c1F Chemical compound Cc(cc1F)cc(F)c1F UHIGHLGTNVYXOP-UHFFFAOYSA-N 0.000 description 1
- WJIFKOVZNJTSGO-UHFFFAOYSA-N Cc1cc(Br)ccc1 Chemical compound Cc1cc(Br)ccc1 WJIFKOVZNJTSGO-UHFFFAOYSA-N 0.000 description 1
- ZLSCQTGTUTYYFG-UHFFFAOYSA-N Cc1cc(OCc2ccc(C(F)(F)F)cc2)ccc1 Chemical compound Cc1cc(OCc2ccc(C(F)(F)F)cc2)ccc1 ZLSCQTGTUTYYFG-UHFFFAOYSA-N 0.000 description 1
- OVWYEQOVUDKZNU-UHFFFAOYSA-N Cc1cccc(C=O)c1 Chemical compound Cc1cccc(C=O)c1 OVWYEQOVUDKZNU-UHFFFAOYSA-N 0.000 description 1
- RLSSMJSEOOYNOY-UHFFFAOYSA-N Cc1cccc(O)c1 Chemical compound Cc1cccc(O)c1 RLSSMJSEOOYNOY-UHFFFAOYSA-N 0.000 description 1
- YXFVVABEGXRONW-UHFFFAOYSA-N Cc1ccccc1 Chemical compound Cc1ccccc1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
- C07D495/04—Ortho-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
- C07D495/14—Ortho-condensed systems
Abstract
The present invention relates to target the inhibitor of BRD4 and its application in anti-tumor medicine, belong to antitumor pharmaceutical technology.Present invention solves the technical problem that being to provide a kind of compound as BRD4 inhibitor.The compound includes compound or its pharmaceutically acceptable salt as follows.Compound of the invention or its pharmaceutically acceptable salt, can be as BRD4 inhibitor, with obvious anti-breast cancer therapeutic effect.
Description
Technical field
Application the present invention relates to BRD4 inhibitor and its in anti-tumor medicine, belongs to anti-tumor medicine and finds skill
Art field.
Background technology
Bromine (BRD4) containing protein 4 is the member of bromine and extra terminal (BET) protein family, in cell division and transcription
Regulation and control transmit epigenetic memory as a central location.Except BRD4, also other three members, such as BRD2, BRD3 and
BRDT, they may regulate and control many cell processes by the interaction between bromine domain.BRD4 is a kind of core of wide expression
Albumen, molecular weight is 200kDa, is in structure conservative but with not containing two bromine domains (BD1 and BD2) of series connection
Identical function and an ET domain.Recently, BRD4 is reported and participates in oncogene rearrangement, causes height oncogenic fusion egg
In vain, so as to play crucial effect, such as breast cancer in the development of the cancer of many types.BRD4 regulation breast cancer cell transfers
It is by adjusting active and regulation ER α inductions the gene expression of Sipa1 enzymes by influenceing the rna plymerase ii and group of breast cancer
The mono- ubiquitinations of albumen H2B extend related phosphorylation.Additionally, BRD4 adjusts the expression of extracellular matrix, in branch prediction
Often occur in gene signal；And progress and/or the existence of breast cancer can be predicted by its activation signal.
Autophagy is conservative, the lysosomal degradation process of multi-step in a kind of evolution, for removing impaired or unnecessary egg
White matter and organelle.Autophagy machine includes that more than 36 autophagy related genes (ATGs), and UNC-51- samples kinases 1 (ULK1) it is
The direct homologous gene of saccharomycete Atg1, is unique serine threonine kinases, is the startup in these autophagy related genes
Enzyme.The complexity of ULK is mutually fit, including ULK1, mAtg13, FIP200 and Atg101, and these are necessary to starting autophagy process
[17].In autophagy, mTORC1 and AMPK can carry out negative regulation by Direct Phosphorylation to ULK1 activity.AMPK-mTORC1-
ULK1 axles may play key effect in the formation of autophagosome and the regulation and control of autophagy.The function and its ginseng of AMPK mTORC1, ULK1
With many pathophysiological processes such as breast cancer be elucidated.Autophagy is a kind of physiological mechanism, can be as temporary transient existence
Means, thus it is referred to as protectiveness cell autophagy.If conversely, cellular stress causes the autophagy of continuous or excessive induction, cell
The related cell death (ACD) of dead or cell autophagy will come one after another.Target the thin of the autophagy correlation that small-molecule drug is induced
Born of the same parents are dead now as a promising strategy.Therefore, the related cell of targeting AMPK-mTORC1-ULK1 modulation autophagy is dead
It is the new way of breast cancer treatment to die.
Recently, (+) JQ1 is verified as first BRD4 inhibitor in terms of protectiveness cell autophagy.Example
Such as, in acute myeloid leukemia (AML), the protection autophagy that AMPK-ULK1 is adjusted generates resistance to BRD4 inhibitor (+) JQ1
The property of medicine.(+) JQ1 has also been reported and has suppressed to protect cell autophagy to be expressed in AML cells so as to increase NPM1 and HEXIM1.Can
See, long-term prescription can lead oncogenic recurrence and transfer with the drug resistance of tumour cell.Although (+) JQ1 has good work
Property, but it lacks selectivity, is a kind of inhibitor of general BET families, it is impossible to the suppression BRD4 of enough selectivitys.Additionally, (+)
The internal pharmacodynamic property extreme difference of JQ1, druggability is very low, hinders it and enters clinical test.Accordingly, it would be desirable to develop one kind
New BRD4 inhibitor.
The content of the invention
Present invention solves the technical problem that being to provide a kind of noval chemical compound as BRD4 inhibitor.
The present invention provides compound or its pharmaceutically acceptable salt of the structural formula as shown in formula I：
Wherein, R1And R2It independently is hydrogen or C1-C4 alkyl, and R1And R2It is asynchronously hydrogen；Or R1And R2Connection is formed
Cyclic structure, the cyclic structure is
R3It is phenyl ring or substituted phenyl ring；
N is 0 or 1.
The present invention also provides the use of above-claimed cpd or its pharmaceutically acceptable salt in treatment breast cancer medicines are prepared
On the way.
Further, the treatment breast cancer medicines are preferably BRD4 inhibitor class medicines.
The present invention a kind of pharmaceutical composition for treating breast cancer is also provided, it comprising effective dose above-claimed cpd or its
Pharmaceutically acceptable salt.
Compound of the invention or its pharmaceutically acceptable salt, can be as BRD4 inhibitor, with significantly anti-
Breast cancer treatment effect.Particularly compound 36, is capable of being combined with the BD1 domains of BRD4 for selectivity, in breast cancer it
The autophagy cell death that can in vivo and in vitro induce BRD4-AMPK to adjust, this is first and targets BRD4 while be reported
With BRD4-AMPK interact micromolecular inhibitor, will be treat breast cancer treatment in, exploitation have new structure and
The BRD4 inhibitor of PPIs effects provides an effective example.
Brief description of the drawings
Fig. 1 be respectively with treatment MCF-7 and MDA-MB-231 the cell certain hour of 1.5 or 3 μM of FL-411 after, then
Cell detects MDC fluorescent staining pictures with fluorescence microscope.
Fig. 2 be respectively with treatment MCF-7 and MDA-MB-231 the cell certain hour of 1.5 or 3 μM of FL-411 after, then
With the picture of Electron microscopy.
Fig. 3 is with GFP-LC3 plasmid transfection MCF-7 and MDA-MB-231 cells, then with 1.5 or 3 μM of FL-411's
Treatment, the picture detected under fluorescence microscope.
Fig. 4 is treatment MCF-7 and MDA-MB-231 cell 24 hours respectively with 1.5 or 3 μM of FL-411, by immune
Groupization detects the expression of LC3B.
Fig. 5 is respectively, with after treatment MCF-7 and MDA-MB-231 the cell certain hour of FL-411, to be examined with Western blot
Survey the expression of Beclin-1, p62 and LC3.
Fig. 6 be in the presence of BafA1, with treatment MCF-7 and the MDA-MB-231 cell 24 hours for using FL-411 respectively,
The expression of p62 and LC3 is detected with Western blot.
Fig. 7 be respectively with treatment MCF-7 and MDA-MB-231 the cell certain hour of 1.5 or 3 μM of FL-411 after, then use
3-MA is added before FL-411 treatment, then cell survival rate is detected with MTT.
Fig. 8 be respectively with treatment MCF-7 and MDA-MB-231 the cell certain hour of 1.5 or 3 μM of FL-411 after, with exempting from
Epidemic disease blotting detects AKT, the expression quantity of p-AKT, mTOR, p-mTOR, p70S6K and p-p70S6K.
Fig. 9 be respectively with treatment MCF-7 and MDA-MB-231 the cell certain hour of 1.5 or 3 μM of FL-411 after, with exempting from
Epidemic disease blotting detects ULK1, the expression quantity of p-ULK1, ATG13, p-ATG13, FIP200 and ATG101.
Figure 10 is thin with siRNA and negative control siRNA the transfection MCF-7 and MDA-MB-231 of two species specificity BRD4
Born of the same parents, BRD4, the expression of c-Myc, AMPK α, p-AMPK α, ULK1, p-ULK1, p62 and LC3 are detected with Western blot.
Figure 11 is the mouse for being inoculated with MCF-7 or MDA-MB-231 daily with the FL-411 treatment of blank or various concentrations
Once.Gross tumor volume is represented with standard error (mean ± SEM).
Figure 12 is the tumor volume change for analyzing last day.*p<0.05,***p<0.001, made comparisons with control group.
Figure 13 is the tumor weight of each group mouse.*p<0.05,***p<0.001, made comparisons with control group.
Figure 14 is the changes of weight of mouse during FL-411 is processed.
Figure 15 is the proliferation marker KI67 and autophagy mark LC3II in immunohistochemical analysis mouse tumor.Quantitative point
Analyse the percentage of each group picture positive staining.Tumour takes from control and FL-411 (100mg/kg) treatment group mouse.***p<
0.001, made comparisons with control group.Scale is 200 μm.
Figure 16 is the tumor tissues for taking MCF-7 and MDA-MB-231 xenograft mouses, cracking.Western blot are analyzed
The expression of BRD4, c-Myc, LC3, and caspase3.
Figure 17 is antitumor vigor of the FL-411 in zebra fish model.CM-DIL mark MCF7 cells add blank or
12.5,25,50 μM of FL-411 treatment, bright field and fluorescence picture are caught with fluorescence microscope.
Figure 18 is tumour fluorescence intensive analysis (mean ± SEM) and each concentration FL-411 treatment 48h in zebra fish model
Inhibiting rate (* * p of each group gross tumor volume afterwards<0.01,***p<0.001, made comparisons with control group).
Specific embodiment
The present invention provides the compound as shown in formula I：
Wherein, R1And R2It independently is hydrogen or C1-C4 alkyl, and R1And R2It is asynchronously hydrogen；Or R1And R2Connection is formed
Cyclic structure, the cyclic structure isR3It is phenyl ring or substituted
Phenyl ring；N is 0 or 1.
Used as one of which implementation method, n is 0；R1And R2It independently is hydrogen or C1-C4 alkyl, and R1And R2It is asynchronously
Hydrogen；Or R1And R2Connection forms cyclic structure, and the cyclic structure is R3It is phenyl ring or substituted phenyl ring.
Below for n be 0 it is several preferred embodiment.
One kind is R1And R2Connection is formedR3It is phenyl ring or substituted phenyl ring.
It is preferred that R1And R2Connection is formed
More preferably R1And R2Connection is formedR3For
Second is R1And R2It is preferably independently hydrogen or C1-C4 alkyl, and R1And R2It is asynchronously hydrogen；R3It is phenyl ring or quilt
Substituted phenyl ring.
It is preferred that R1It is hydrogen；R2It is C1-C4 alkyl；R3For
More preferably R1It is hydrogen；R2ForR3For
Further preferred R1It is hydrogen；R2ForR3For
The third is R1And R2Connection is formedR3It is phenyl ring or substituted phenyl ring.
More preferably R1And R2Connection is formedR3For
4th kind is R1And R2Connection is formedR3It is phenyl ring or substituted phenyl ring.
It is preferred that R1And R2Connection is formedR3For
More preferably R1And R2Connection is formedR3For
Also a kind of is R1And R2Connection is formedR3It is phenyl ring or substituted phenyl ring.
It is preferred that R1And R2Connection is formedR3For
Used as another embodiment, n is 1；R1And R2It independently is hydrogen or C1-C4 alkyl, and R1And R2It is asynchronously
Hydrogen；Or R1And R2Connection forms cyclic structure, and the cyclic structure is
R3It is phenyl ring or substituted phenyl ring.
Preferably, n is 1, R1And R2Connection is formedR3For
More preferably n is 1, R1And R2Connection is formedR3For
Here is some preferred structures of compound of the invention.
The present invention also provides the pharmaceutically acceptable salt of compound of the present invention.The salt can for nitrate,
Hydrochloride, sulfate or phosphate etc..
The present invention also provides the use of above-claimed cpd or its pharmaceutically acceptable salt in treatment breast cancer medicines are prepared
On the way.Further, the treatment breast cancer medicines are preferably BRD4 inhibitor class medicines, for the associated treatment of breast cancer.
The present invention a kind of pharmaceutical composition for treating breast cancer is also provided, it be the above-claimed cpd comprising effective dose or
The preparation of its pharmaceutically acceptable salt.The compounds of this invention can be made following shape by methods known in the art
Formula：Tablet, capsule, aqueous or oily solutions, supensoid agent, emulsion, cream, ointment, gel, nasal spray, suppository,
For suck tiny scattered pulvis or aerosol or spray, for parenteral (including intravenous, intramuscular or infusion)
Sterile aqueous or oily solution or supensoid agent or without bacterial emulsion.Can using sterilized water or water-propylene glycol solution as solvent come
Liquid preparation is prepared, can also be by active ingredients in Aqueous Solutions of Polyethylene Glycol.Aqueous solution for orally giving can lead to
Cross and active component is dissolved in water and adds suitable colouring agent, flavouring, stabilizer and thickener to prepare on demand.Mouthful
The aqueous suspension that clothes are used can be dispersed in water by by tiny scattered active component together with stickum, the viscosity
Material is as being natural synthetic gums, resin, methylcellulose, carboxymethylcellulose calcium and suspending agent known to other pharmaceutical arts.
Pharmaceutical composition can be unit dosage form.In these forms, the composition is divided into containing appropriate activearm
The UD for dividing.The unit dosage form can be packaged preparation, and packaging includes the preparation of separation amount, such as box-packed tablet,
Capsule and the pulvis in phial or ampoule.Unit dosage form can also for capsule, cachet or tablet or its can be
An appropriate number of any these packaged forms.
Pharmaceutical composition of the invention, its active component can be only compound of the invention, also can be with other anti-breast cancers
Compound group cooperation is active component.
During breast cancer is treated, can be combined with other anti-breast cancer medicines using pharmaceutical composition of the invention
Treatment.
Treat breast cancer when, can by simultaneously, it is sequential or individually give various therapeutic components can realize it is this combine control
Treat.Other pharmaceutical actives in the compounds of this invention and allowance dosage range in such combination product application effective dosage ranges
Agent.
Specific embodiment of the invention is further described with reference to embodiment, is not therefore limited the present invention
System is among described scope of embodiments.
The synthesis of the compound 1~42 of embodiment 1
The logical method of the synthesis of intermediate 2a-e：
Take ketone or aldehyde (30.0mmol), cyan-acetic ester (30.0mmol), elemental sulfur (30.0mmol) be dissolved in 200ml without
In water-ethanol, triethylamine (35.0mmol) .The resulting was allowed to refluxed for are added
12h. reactant mixtures are warming up to backflow, insulation reaction 12 hours or so.After completion of the reaction, room temperature filtering is cooled to, with anhydrous
Ethanol washs (30ml x 3), merging filtrate removal of solvent under reduced pressure, and crude product obtains intermediate by silica gel column chromatography purifying, pale yellow
Color solid.
Intermediate 2a1H-NMR(400MHz,CDCl3-d6),δ(ppm):5.73 (1H, s), 4.18 (2H, q, J=
7.1Hz), 2.47 (2H, m), 1.53 (2H, m), 1.25 (3H, t, J=7.0Hz), 0.87 (3H, t, J=7.2Hz) .HRMS
(ESI)+calculated forC10H15NO2S,[M+H]+:m/z 214.0902,found 214.0910.
Intermediate 2b1H-NMR(400MHz,CDCl3),δ(ppm):5.85(2H,s),4.24(2H,m),2.84(2H,m),
2.72 (2H, m), 2,30 (2H, m), 1.32 (3H, t, J=7.1Hz)；HRMS(ESI)+calculated for C10H14NO2S,
[M+H]+:m/z 212.0745,found 212.0752.
Intermediate 2c1H-NMR(400MHz,CDCl3),δ(ppm):5.73 (2H, s), 4.26 (2H, q, J=6.9Hz),
2.94 (2H, t, J=5.3Hz), 2.55 (2H, t, J=5.3Hz), 1.79 (2H, m), 1.60 (4H, m), 1.32 (3H, t, J=
7.2Hz).
Intermediate 2d1H-NMR(400MHz,DMSO-d6),δ(ppm):5.92 (2H, s), 4.26 (2H, q, J=14.2,
7.13Hz), 2.87 (1H, m), 2.56 (2H, m), 2.12 (1H, m), 1.81 (2H, m), 1.33 (4H, t, J=7.1Hz), 1.04
(3H, d, J=6.5Hz)；HRMS(ESI)+calculated for C12H18NO2S,[M+H]+:m/z 240.1058,found
240.1067.
Intermediate 2e1H-NMR(400MHz,CDCl3),δ(ppm):5.98(2H,s),4.26(2H,m),3.37(2H,t,J
=2.0Hz), 2.84 (2H, m), 2.66 (2H, t, J=5.9Hz), 2,44 (3H, s), 1.33 (3H, t, J=7.1Hz)
The logical method of the synthesis of compound 1~42：
Intermediate obtained in step (2.0mmol) is taken respectively to be dissolved in the anhydrous dioxane of 3ml hydrogen chloride gas saturations, then is added
Enter carbonitrile derivatives (2.2mmol).Mixture is heated to 100 DEG C insulation reaction 6-12 hours.Room temperature is cooled to after completion of the reaction, is used
The sodium bicarbonate aqueous solution quenching reaction of 20ml saturations.The off-white powder for the solid of precipitation being collected by filtration and being washed with methyl alcohol is produced
Thing.
Reaction reagent and reaction condition：(a)NCCH2CO2Et,S8,EtOH,Et3N, backflow, 12h；(b) nitrile, dioxane/
HCl,100℃,12h。
The following is nuclear-magnetism result：
Compound 1
1H-NMR(400MHz,DMSO-d6),δ(ppm):7.85 (2H, m), 7.25 (1H, t, J=7.9Hz), 6.88 (1H,
Ddd, J=8.4,2.7,1.0Hz), 6.78 (1H, br s), 3.80 (3H, s), 2.71 (2H, t, J=7.3Hz), 1.65 (2H,
M), 0.94 (3H, t, J=7.3Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):160.6,155.2,150.5,149.1,
131.7,124.3,123.9,123.0,120.4,118.4,118.0,112.4,55.1,31.6,23.6,11.4；HRMS(ESI
)+calculated for C16H17N2O2S,[M+H]+:m/z 301.1011,found 301.1020.
Compound 2
1H-NMR(400MHz,DMSO-d6),δ(ppm):8.42 (1H, t, J=1.6Hz), 8.26 (1H, dt, J=7.8,
1.3Hz), 7.49 (1H, dq, J=7.8,1.0Hz), 7.32 (1H, t, J=7.8Hz), 6.79 (1H, br s), 2.71 (2H, t, J
=7.0Hz), 1.65 (2H, m), 0.94 (3H, t, J=7.3Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):159.5,
156.6,153.2,151.8,141.5,133.8,133.0,128.5,127.6,126.8,126.5,120.4,34.0,26.0,
13.8；HRMS(ESI)+calculated for C15H14BrN2OS,[M+H]+:m/z 349.0010,found 349.0017.
Compound 3
1H-NMR(400MHz,DMSO-d6),δ(ppm):8.09 (2H, d, J=8.3Hz), 6.72 (2H, t, J=8.2Hz),
2.69 (2H, t, J=7.4Hz), 1.63 (2H, q, J=14.4,7.1Hz), 0.93 (3H, t, J=7.5Hz)；13C-NMR
(100MHz,CF3COOD),δ(ppm):159.5,156.6,153.2,151.8,141.5,133.8,133.0,128.5,
127.6,124.8,113.8,111.6,34.0,26.0,13.8；HRMS(ESI)+calculated for C15H15N2O2S,[M+
H]+:m/z 287.0854,found 287.0863.
Compound 4
1H-NMR(400MHz,DMSO-d6),δ(ppm):7.62 (2H, dd, J=8.4,1.3Hz), 7.55 (2H, d, J=
8.4Hz), 7.41 (4H, m), 7.32 (1H, m), 6.81 (1H, s), 3.80 (2H, s), 2.70 (2H, t, J=7.0Hz), 1.61
(1H, m), 0.91 (3H, t, J=7.5Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):162.1,152.6,152.6,
150.7,145.6,141.5,132.3,132.3,130.8,130.8,130.6,130.6,130.0,128.8,128.8,
126.5,120.3,120.3,39.8,33.9,26.1,13.9；HRMS(ESI)+calculated for C22H21N2OS,[M+H
]+:m/z 361.1375,found 361.1381.
Reaction reagent and reaction condition：(a)NCCH2CO2Et,S8,EtOH,Et3N, backflow, 12h；(b) nitrile, dioxane/
HCl,100℃,12h。
Compound 5
1H-NMR(400MHz,DMSO-d6),δ(ppm):12.60 (1H, s), 7.71 (2H, m), 7.45 (1H, dd, J=
), 8.8,6.1Hz 7.13 (1H, d, J=6.1Hz), 3.86 (3H, s), 2.41 (4H, m), 2.95 (2H, m)；13C-NMR
(100MHz,CF3COOD),δ(ppm):161.6,155.5,151.2,150.7,146.4,142.7,132.7,125.5,
124.0,121.4,113.2,112.6,56.1,29.4,28.6,28.4；HRMS(ESI)+calculated for
C16H15N2O2S,[M+H]+:m/z 299.0854,found 299.0860.
Compound 6
1H-NMR(400MHz,DMSO-d6),δ(ppm):12.83 (1H, s), 8.33 (2H, d, J=8.1Hz), 7.90
(2H, d, J=8.1Hz), 2.96 (4H, m), 2.95 (2H, m)；13C-NMR(100MHz,CF3COOD),δ(ppm):159.7,
157.3,153.1,151.8,136.4,136.4,135.2,135.2,131.3,131.3,126.4,124.5,120.6,34.2,
26.2,14.0；HRMS(ESI)+calculated for C16H12F3N2OS,[M+H]+:m/z 337.0622,found
337.0630.
Compound 71H-NMR(400MHz,DMSO-d6),δ(ppm):12.69 (1H, s), 8.34 (1H, dd, J=1.8,
1.7Hz), 8.12 (1H, d, J=8.0Hz), 7.73 (1H, d, J=7.8Hz), 7.47 (1H, dd, J=8.0,7.8Hz), 2.93
(4H,m),2.40(2H,m)；13C-NMR(100MHz,CF3COOD),δ(ppm):161.3,159.2,155.5,152.1,
151.6,130.3,130.3,129.1,129.1,128.8,125.8,116.4,30.4,29.5,29.3；HRMS(ESI)+
calculated for C15H12BrN2OS,[M+H]+:m/z 346.9854,found 346.9863.
Compound 81H-NMR(400MHz,DMSO-d6),δ(ppm):12.54(1H,s),7.32(4H,m),7.25(1H,
m),3.93(2H,s),2.88(4H,m),2.36(2H,m)；13C-NMR(100MHz,CF3COOD),δ(ppm):168.8,
158.7,156.2,139.8,137.2,137.0,129.3,129.3,129.0,129.0,127.3,118.6,29.4,29.1,
27.9；HRMS(ESI)+calculated for C16H15N2OS,[M+H]+:m/z 283.0905,found 283.0914.
Compound 91H-NMR(400MHz,DMSO-d6),δ(ppm):12.70 (1H, s), 8.07 (2H, d J=
8.6Hz), 7.73 (2H, d J=8.6Hz), 2.95 (4H, m), 2.41 (2H, m)；13C-NMR(100MHz,CF3COOD),δ
(ppm):161.3,159.2,155.5,152.1,151.6,130.3,130.3,129.1,129.1,128.8,125.8,
116.4,30.4,29.5,29.3；HRMS(ESI)+calculated for C15H12BrN2OS,[M+H]+:m/z 346.9854,
found 346.9864.
Compound 101H-NMR(400MHz,DMSO-d6),δ(ppm):12.22(1H,s),8.99(1H,s),7.81
(2H,s),2.92(4H,m),2.39(2H,m),2.23(6H,s)；13C-NMR(100MHz,CF3COOD),δ(ppm):162.8,
156.7,155.6,146.4,144.0,136.5,131.1,130.9,130.9,129.6,125.0,116.5,30.8,30.1,
29.9,16.5,16.5；HRMS(ESI)+calculated for C17H17N2O2S,[M+H]+:m/z 313.1011,found
313.1020.
Compound 111H-NMR(400MHz,DMSO-d6),δ(ppm):12.61(1H,s),8.13(2H,m),7.55
(3H,m),2.96(4H,m),2.41(2H,m)；13C-NMR(100MHz,CF3COOD),δ(ppm):162.9,157.3,151.8,
133.6,133.6,129.3,129.3,128.1,128.1,126.9,121.6,117.0,29.8,29.0,28.8；HRMS
(ESI)+calculated for C15H13N2OS,[M+H]+:m/z 269.0749,found 269.0753.
Compound 121H-NMR(400MHz,DMSO-d6),δ(ppm):12.73 (1H, s), 8.32 (2H, d, J=
8.0Hz), 7.90 (2H, d, J=8.0Hz), 3.28 (2H, m), 2.87 (2H, m), 1.87 (2H, m), 1.60 (4H, m)；13C-NMR
(100MHz,CF3COOD),δ(ppm):160.9,157.8,153.3,146.7,144.7,138.9,138.0,,128.3,
127.1,126.5,123.8,122.8,121.1,31.3,29.2,26.8,26.8,25.8；HRMS(ESI)+calculated
for C18H16F3N2OS,[M+H]+:M/z 365.0935, found 365.0942. compounds 131H-NMR(400MHz,DMSO-
d6),δ(ppm):8.33 (1H, m), 8.15 (1H, d, J=7.8Hz), 7.70 (1H, d, J=7.8Hz), 7.44 (1H, t, J=
7.8Hz),3.29(2H,m),2.83(2H,m),1.85(2H,m),1.61(4H,m)；13C-NMR(100MHz,CF3COOD),δ
(ppm):160.1,155.6,149.0,146.7,141.4,141.2,133.8,132.8,128.4,127.3,126.8,
125.0,33.7,31.6,29.2,28.8,28.1；HRMS(ESI)+calculated for C17H16BrN2OS,[M+H]+:m/
z375.0167,found 375.0174.
Compound 141H-NMR(400MHz,DMSO-d6),δ(ppm):12.39(1H,s),9.71(1H,s),7.52(2H,
M), 7.30 (1H, t, J=7.5Hz), 6.95 (1H, d, J=6.7Hz), 3.29 (2H, br s), 2.84 (2H, br s), 1.85
(2H,br s),1.64(4H,m)；13C-NMR(100MHz,CF3COOD),δ(ppm):160.6,158.3,158.3,156.4,
149.0,146.3,141.0,134.2,126.8,125.8,123.0,116.5,33.6,31.4,29.1,28.7,28.1；HRMS
(ESI)+calculated for C17H17N2O2S,[M+H]+:m/z313.1011,found 313.1020.
Compound 151H-NMR(400MHz,DMSO-d6),δ(ppm):12.20(1H,s),10.04(1H,s),8.00
(2H, d, J=8.8Hz), 6.86 (2H, d, J=8.8Hz), 3.27 (2H, m), 2.82 (2H, m), 1.85 (2H, br s), 1.62
(4H,m)；13C-NMR(100MHz,CF3COOD),δ(ppm):160.4,157.6,155.8,149.3,147.0,141.4,
132.5,132.5,125.3,125.3,125.1,124.9,34.0,31.8,29.4,29.0,28.4；HRMS(ESI)+
calculated for C17H17N2O2S,[M+H]+:m/z 313.1011,found 313.1019.
Compound 161H-NMR(400MHz,DMSO-d6),δ(ppm):12.61 (1H, s), 8.07 (2H, d, J=
8.1Hz), 7,75 (2H, d, J=8.1Hz), 3.32 (2H, m), 2.88 (2H, m), 1.89 (2H, br s), 1.65 (4H, m)；13C-
NMR(100MHz,CF3COOD),δ(ppm):160.5,156.3,152.7,146.6,141.3,136.3,136.3,135.0,
131.2,131.2,124.9,116.9,33.9,31.7,29.3,29.0,28.3；HRMS(ESI)+calculated for
C17H16BrN2OS,[M+H]+:m/z 375.0167,found 375.0174.
Compound 171H-NMR(400MHz,DMSO-d6),δ(ppm):12.11(1H,s),8.97(1H,s),7.80
(2H,s),3.28(2H,m),2.83(2H,m),2.23(6H,s),1.86(2H,br s),1.63(4H,m)；13C-NMR
(100MHz,CF3COOD),δ(ppm):162.6,156.2,152.6,149.6,145.0,140.9,136.3,130.9,
130.8,130.8,129.5,129.5,33.9,31.5,29.3,28.9,28.3,16.4,16.4；HRMS(ESI)+
calculated for C19H21N2O2S,[M+H]+:m/z 341.1324,found 341.1331.
Compound 181H-NMR(400MHz,DMSO-d6),δ(ppm):12.52(1H,s),7.41-7.29(4H,m),
7.25(1H,br s)3.91(2H,s),3.23(2H,br s),2.80(2H,br s),1.84(2H,s),1.59(4H,m)；13C-
NMR(100MHz,CF3COOD),δ(ppm):160.8,152.4,151.9,131.9,131.9,131.8,131.8,131.6,
131.6,129.8,129.8,116.6,39.7,33.6,31.1,29.0,28.7,28.0；HRMS(ESI)+calculated
for C18H19N2OS,[M+H]+:m/z 311.1218,found 311.1223.
Compound 191H-NMR(400MHz,DMSO-d6),δ(ppm):7.63(4H,m),7.44(4H,m),7.34(1H,
m),3.92(2H,s),3.23(2H,m),2.77(2H,m),1.82(2H,m),1.56(4H,m)；13C-NMR(100MHz,
CF3COOD),δ(ppm):160.2,158.6,157.4,145.9,143.6,143.1,139.2,138.5,129.8,129.8,
128.4,128.4,128.2,128.2,127.6,126.4,126.4,122.5,37.2,31.4,29.0,26.8,26.5,
25.8；HRMS(ESI)+calculated for C24H23N2OS,[M+H]+:m/z 387.1531,found 387.1540.
Reaction reagent and condition：(a)NCCH2CO2Et,S8,EtOH,Et3N, backflow, 12h；(b) nitrile, dioxane/HCl,
100℃,12h。
Compound 201H-NMR(400MHz,DMSO-d6),δ(ppm):12.51 (1H, s), 7.73 (1H, d, J=
7.9Hz), 7.68 (1H, t, J=1.9Hz), 7.42 (1H, t, J=8.2Hz), 7.13 (1H, dq, J=8.2,0.7Hz), 3.85
(3H, s), 3.13 (1H, m), 2.87 (1H, dd, J=16.5,4.9Hz), 2.76 (1H, m), 2.36 (1H, m), 1.89 (2H, m),
1.41 (1H, m), 1.07 (3H, d, J=6.6Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):13C-NMR(100MHz,
CF3COOD),δ(ppm):163.0,160.0,157.0,151.2,142.0,135.1,134.1,126.7,125.4,124.5,
122.8,114.6,57.5,34.5,31.3,31.1,26.5,21.6；HRMS(ESI)+calculated for C18H19N2O2S,
[M+H]+:m/z 327.1167,found 327.1170.
Compound 211H-NMR(400MHz,DMSO-d6),δ(ppm):12.74 (1H, s), 8.33 (2H, d, J=8.4Hz),
7.90 (2H, d, J=8.4Hz), 3.13 (1H, m), 2.90 (1H, dd, J=17.8,5.5Hz), 2.83 (1H, m), 2.40 (1H,
M), 1.90 (2H, m), 1.40 (1H, m), 1.07 (3H, d, J=6.5Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):
159.9,156.3,151.4,142.8,135.3,130.9,130.9,129.7,129.6,129.2,126.3,125.0,
123.6,34.7,31.4,31.1,26.5,21.7；HRMS(ESI)+calculated for C18H16F3N2OS,[M+H]+:m/z
365.0935,found 365.0941.
Compound 221H-NMR(400MHz,DMSO-d6),δ(ppm):12.60(1H,s),8.31(1H,s),8.13(1H,
D, J=8.0Hz), 7.77 (1H, d, J=7.9Hz), 7.48 (1H, t, J=8.0Hz), 3.13 (1H, m), 2.88 (1H, dd, J=
17.2,4.7Hz), 2.80 (1H, m), 2.38 1H, m), 1.90 (2H, m), 1.38 (1H, m), 1.07 (3H, d, J=6.4Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):161.0,156.0,151.2,142.4,141.5,141.5,135.1,
133.9,133.9,133.0,128.5,127.4,126.8,34.5,31.3,31.0,26.5,21.6；HRMS(ESI)+
calculated for C17H16BrN2OS,[M+H]+:m/z 375.0167,found 375.0178.
Compound 231H-NMR(400MHz,DMSO-d6),δ(ppm):12.65(1H,m),8.10(2H,m),3.13(1H,
M), 2.89 (1H, m), 2.78 (1H, m), 2.39 (1H, m), 1.89 (2H, m), 1.38 (1H, m), 1.07 (3H, d, J=
6.7Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):159.9,156.2,154.5,153.7,151.8,148.8,
146.1,142.9,135.2,124.9,122.1,115.8,34.7,31.4,31.1,26.6,21.7；HRMS(ESI)+
calculated for C17H14F3N2OS,[M+H]+:m/z 351.0779,found 351.0087.
Compound 241H-NMR(400MHz,DMSO-d6),δ(ppm):12.4(1H,s),9.73(1H,s),7.55(2H,
M), 7.30 (1H, t, J=7.7Hz), 6.95 (1H, ddd, J=8.1,2.4,0.8Hz), 3.12 (1H, m), 2.83 (1H, dd, J
=16.6,4.7Hz), 2.75 (1H, m), 2.36 (1H, m), 1.87 (2H, m), 1.38 (1H, m), 1.06 (3H, d, J=
6.6Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):159.9,158.6,157.2,151.2,142.2,135.2,
134.5,127.1,126.1,124.7,123.3,116.7,34.7,31.4,31.2,26.6,21.7；HRMS(ESI)+
calculated for C17H17N2O2S,[M+H]+:m/z 313.1011,found 313.1020.
Compound 251H-NMR(400MHz,DMSO-d6),δ(ppm):12.25(1H,s),10.15(1H,s),8.01
(2H, d, J=8.7Hz), 6.87 (2H, d, J=8.7Hz), 6.87 (2H, d, J=8.7Hz), 3.11 (1H, m), 2.84 (1H,
M), 2.75 (1H, m), 2.35 (1H, m), 1.88 (2H, m), 1.40 (1H, m), 1.06 (3H, d, J=6.5Hz)；13C-NMR
(100MHz,CF3COOD),δ(ppm):165.2,160.0,156.5,151.3,141.1,134.9,132.6,132.6,
123.7,119.9,119.9,117.4,34.4,31.3,31.1,26.4,21.6；HRMS(ESI)+calculated for
C17H17N2O2S,[M+H]+:m/z 313.1011,found 313.1022.
Compound 261H-NMR(400MHz,DMSO-d6),δ(ppm):12.59 (1H, s), 8.07 (2H, d, J=
8.5Hz), 7.73 (2H, dt, J=8.7,1.9Hz), 3.13 (1H, m), 2.88 (1H, dd, J=16.6,4.8Hz), 2.78 (1H,
M), 2.33 (1H, m), 1.88 (2H, m), 1.39 (1H, m), 1.07 (3H, d, J=6.6Hz)；13C-NMR(100MHz,
CF3COOD),δ(ppm):160.3,156.7,151.5,142.2,136.4,136.4,135.2,135.1,131.2,131.2,
124.6,124.3,34.6,31.4,31.1,26.5,21.7；HRMS(ESI)+calculated for C17H16BrN2OS,[M+
H]+:m/z 375.0088,found 375.0093.
Compound 271H-NMR(400MHz,DMSO-d6),δ(ppm):12.12(1H,s),8.97(1H,s),7.79(2H,
S), 3.10 (1H, m), 2.83 (1H, dd, J=16.6,4.6Hz), 2.73 (1H, m), 2.34 (1H, m), 2.22 (6H, s), 1.87
(2H, m), 1.37 (1H, m), 1.06 (3H, d, J=6.5Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):162.7,
160.2,156.6,151.5,140.7,134.9,130.9,130.9,129.6,129.6,123.5,116.4,34.5,31.4,
31.1,26.5,21.6,16.4,16.4；HRMS(ESI)+calculated for C19H21N2O2S,[M+H]+:m/z
341.1324,found 341.1330.
Compound 281H-NMR(400MHz,DMSO-d6),δ(ppm):12.57(1H,s),9.78(1H,s),7.52(2H,
M), 7.32 (1H, t, J=7.9Hz), 6.98 (1H, ddd, J=8.1,2.4,0.6Hz), 4.03 (2H, s), 3.11 (4H, s),
2.66(3H,s)；13C-NMR(100MHz,CF3COOD),δ(ppm):159.6,158.8,154.4,134.6,134.6,131.5,
129.6,127.1,126.7,123.7,123.4,117.1,54.4,54.1,45.3,45.3,24.6；HRMS(ESI)+
calculated for C17H17N2OS,[M+H]+:m/z 297.1062,found 297.1071.
Compound 291H-NMR(400MHz,DMSO-d6),δ(ppm):12.45(1H,s),7.31(4H,m),7.24(1H,
M), 3.90 (2H, s), 3.01 (1H, m), 2.77 (1H, dd, J=16.8,4.8Hz), 2.69 (1H, m), 2.31 (1H, m), 1.84
(2H, m), 1.34 (1H, m), 1.03 (3H, d, J=6.6Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):161.8,
159.4,150.5,141.7,134.9,132.3,132.3,132.1,131.9,131.9,130.1,124.7,40.1,34.5,
31.4,31.1,26.5,21.6；HRMS(ESI)+calculated for C18H19N2OS,[M+H]+:m/z 311.1218,
found 311.1222.
Compound 301H-NMR(400MHz,DMSO-d6),δ(ppm):12.50(1H,s),7.64(4H,m),7.45(4H,
M), 7.34 (1H, m), 3.96 (2H, s), 3.06 (1H, m), 2.81 (1H, dd, J=16.6,4.7Hz), 2.71 (1H, m), 2.32
(1H, m), 1.85 (2H, m), 1.35 (1H, m), 1.03 (3H, d, J=6.5Hz)；13C-NMR(100MHz,CF3COOD),δ
(ppm):161.4,159.4,150.5,145.4,141.7,141.6,134.8,132.4,132.4,130.9,130.9,
130.6,130.6,130.1,129.4,128.8,128.8,124.7,39.7,34.5,31.4,31.1,26.5,21.7；HRMS
(ESI)+calculated for C24H23N2OS,[M+H]+:m/z 387.1531,found 387.1540.
Compound 311H-NMR(400MHz,DMSO-d6),δ(ppm):12.57 (1H, s), 7.73 (1H, d, J=
7.8Hz), 7.69 (1H, m), 7.44 (1H, dd, J=8.5,7.8Hz), 7.13 (1H, dd, J=8.5,2.5Hz), 3.85 (3H,
s),3.60(2H,s),2.99(2H,m),2.70(2H,m),2.39(3H,s)；13C-NMR(100MHz,CF3COOD),δ(ppm):
160.1,158.6,156.3,141.8,134.0,133.4,131.4,129.4,128.9,128.4,127.0,123.4,54.5,
54.3,45.4,45.4,24.7；HRMS(ESI)+calculated for C17H18N3O2S,[M+H]+:m/z 328.1120,
found 328.1131.
Compound 321H-NMR(400MHz,DMSO-d6),δ(ppm):12.83 (1H, s), 8.34 (2H, d, J=
8.2Hz), 7.96 (2H, d, J=8.2Hz), 3.61 (2H, s), 2.99 (2H, m), 2.69 (2H, m), 2.39 (3H, s)；13C-NMR
(100MHz,CF3COOD),δ(ppm):160.0,159.8,154.7,134.4,134.4,131.6,129.4,127.0,
126.3,123.3,123.3,115.0,115.0,57.7,54.4,45.4,24.6；HRMS(ESI)+calculated for
C17H15F3N3OS,[M+H]+:m/z 366.0888,found 366.0897.
Compound 331H-NMR(400MHz,DMSO-d6),δ(ppm):12.83 (1H, s), 8.40 (1H, t, J=1.8,
1.7Hz), 8.23 (1H, ddd, J=8.0,1.4,1.1Hz), 7.58 (1H, m), 7.37 (1H, dd, J=8.0,7.8Hz), 3.50
(2H,s),2.96(2H,m),2.63(2H,m),2.37(3H,s)；13C-NMR(100MHz,CF3COOD),δ(ppm):160.7,
158.4,158.4,140.3,140.0,131.1,129.5,129.5,129.3,126.5,123.8,123.3,54.6,54.4,
45.4,24.7；HRMS(ESI)+calculated for C16H15BrN3OS,[M+H]+:m/z 376.0119,found
376.0126.
Compound 341H-NMR(400MHz,DMSO-d6),δ(ppm):12.55 (1H, s), 8.12 (2H, d, J=
8.0Hz), 7.54 (2H, d, J=8.0Hz), 3.12 (2H, s), 2.82 (2H, m), 2.38 (2H, m), 1.90 (3H, s)；13C-NMR
(100MHz,CF3COOD),δ(ppm):160.2,157.5,157.5,151.2,141.9,138.8,135.1,132.7,
132.7,130.0,125.5,124.7,34.5,31.3,26.5,21.6；HRMS(ESI)+calculated for
C16H16N3O2S,[M+H]+:m/z 314.0963,found 314.0973.
Compound 351H-NMR(400MHz,DMSO-d6),δ(ppm):8.05 (2H, d, J=8.7Hz), 6.78 (2H, d,
J=8.7Hz), 3.49 (2H, s), 2.95 (2H, m), 2.63 (2H, m), 2.36 (3H, s)；13C-NMR(100MHz,CF3COOD),
δ(ppm):165.2,160.0,156.5,151.3,141.1,134.9,132.6,132.6,123.7,119.9,119.9,
117.4,34.4,31.3,31.1,26.4；HRMS(ESI)+calculated for C17H18N3O2,[M+H]+:m/z
296.1399,found 296.1402.
Compound 361H-NMR(400MHz,DMSO-d6),δ(ppm):12.18(1H,s),8.99(1H,s),7.83
(2H,s),3.56(2H,s),2.96(2H,m),2.67(2H,m),2.38(3H,s),2.23(6H,s)；13C-NMR(100MHz,
CF3COOD),δ(ppm):160.3,159.9,158.9,154.0,131.5,131.5,131.5,129.9,129.9,128.2,
122.1,115.8,54.4,54.2,45.4,24.5,16.5,16.5；HRMS(ESI)+calculated for C18H20N3O2S,
[M+H]+:m/z 342.1276,found 342.1280.
Compound 371H-NMR(400MHz,DMSO-d6),δ(ppm):12.58(1H,s),7.63(4H,m),7.44
(4H,m),7.35(1H,m),3.97(2H,s),3.52(2H,m),2.91(2H,m),2.67(2H,m)2.35(3H,s)；13C-
NMR(100MHz,CF3COOD),δ(ppm):164.7,159.0,153.5,145.9,141.7,132.6,132.6,131.4,
131.0,131.0,130.8,130.8,130.2,130.2,129.4,129.0,129.0,128.5,54.3,54.1,45.4,
40.3,24.5；HRMS(ESI)+calculated for C23H22N3OS,[M+H]+:m/z 388.1484,found
388.1494.
Compound 381H-NMR(400MHz,DMSO-d6),δ(ppm):12.58(1H,br s),7.74-7.69(2H,
M), 7.45-7.28 (6H, m), 7.13 (1H, d, J=7.0Hz), 3.85 (3H, s), 3.72 (2H, s), 3.64 (2H, s), 2.98
(2H,br s),2.79(2H,br s)；13C-NMR(100MHz,CF3COOD),δ(ppm):162.8,159.1,154.1,
152.1,133.8,133.3,132.3,132.3,131.7,131.7,130.0,128.2,126.5,125.8,122.8,
119.4,116.5,114.3,63.6,57.2,51.7,51.3,24.2；HRMS(ESI)+calculated for
C23H22N3O2S,[M+H]+:m/z 404.1433,found 404.1434.
Compound 391H-NMR(400MHz,DMSO-d6),δ(ppm):12.19(1H,s),8.99(1H,s),7.81
(2H,s),7.39-7.27(5H,m),3.72(2H,s),3.62(2H,s),2.96(2H,br s),2.79(2H,br s),2.22
(6H,s)；13C-NMR(100MHz,CF3COOD),δ(ppm):164.0,157.7,156.6,151.7,131.6,130.6,
130.6,130.0,130.0,129.6,129.3,129.3,127.7,127.7,126.5,126.4,119.9,113.6,61.9,
50.0,49.6,22.4,14.3,14.3；
HRMS(ESI)+calculated for C24H24N3O2S,[M+H]+:m/z 418.1589,found 418.1597.
Compound 401H-NMR(400MHz,DMSO-d6),δ(ppm):12.57 (1H, s), 8.13 (2H, d, J=
7.0Hz),7.59-7.50(3H,m),7.39-7.26(5H,m),3.72(2H,s),3.64(2H,s),2.98(2H,br s),
2.79(2H,br s)；13C-NMR(100MHz,CF3COOD),δ(ppm):162.2,159.2,159.2,153.7,152.1,
139.1,133.3,132.5,132.5,132.3,132.3,131.7,131.7,130.0,130.0,129.3,125.0,
122.7,63.7,51.7,51.4,24.2；HRMS(ESI)+calculated for C22H20N3OS,[M+H]+:m/z
374.1327,found 374.1330.
Compound 411H-NMR(400MHz,DMSO-d6),δ(ppm):12.80 (1H, s), 8.33 (2H, d, J=
8.2Hz), 7.89 (2H, d, J=8.2Hz), 7.39-7.26 (5H, m), 3.72 (2H, s), 3.65 (2H, s), 2.89 (2H, t, J
=5.8Hz), 2.72 (2H, t, J=5.8Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):164.0,157.9,155.7,
155.4,137.5,131.1,130.1,130.1,129.5,129.5,128.8,128.4,128.4,128.2,126.8,
126.0,123.9,121.2,120.6,61.3,49.6,49.3,22.0；HRMS(ESI)+calculated for
C23H19F3N3OS,[M+H]+:m/z 442.1201,found 442.1210.
Compound 421H-NMR(400MHz,DMSO-d6),δ(ppm):8.08 (2H, d, J=8.7Hz), 7.39-7.26
(5H, m), 6.76 (2H, d, J=8.7Hz), 3.68 (2H, s), 3.54 (2H, s), 2.89 (2H, t, J=5.8Hz), 2.72 (2H,
T, J=5.8Hz)；13C-NMR(100MHz,CF3COOD),δ(ppm):164.2,157.4,156.6,151.6,131.5,
131.0,131.0,130.4,130.4,129.8,129.8,129.5,126.5,126.3,120.0,118.7,118.7,
118.0,61.8,49.9,49.8,22.3；HRMS(ESI)+calculated for C22H20N3O2S,[M+H]+:m/z
390.1276,found 390.1280.
BRD4 inhibitory activity and autophagy the rate activity of the compound 1~42 of test example 1
The purpose of this experiment is detection Compounds in vitro BRD4 inhibitory activity of the invention, and the autophagy rate work to cell
Property, it the results are shown in Table 1.
1st, AlphaScreen detections
In 50 millimeters of hepes, 100mM NaCl, 0.1% bovine serum albumin(BSA), pH value is 7.4 to all of reagent dilutions, is added
Plus 0.05%CHAPS allows balance to room temperature.Ligand concentration scope is formulated as 0~100 μM, takes 4 μ L and is added in 384 plates, then
Add 4 μ LBRD4 (1) enzymes (250nM), Room-temperature seal to be incubated 30 minutes, equimolar biotin peptide H-SGRGK is added afterwards
(Ac)GGK(Ac)GLGK-(Ac)GGAK(Ac)RHRK(Biotin)-OH.Under low light condition, sealing is further incubated for 30 minutes
After add 4 μ L Streptavidins (25 μ grams per milliliters) and 4 μ l nickel chelate acceptor beads (25 μ grams per milliliters).Plate paper tinsel is sealed, lucifuge,
Incubation at room temperature 60min, upper plate reader is read (PHERAstar FS BMG LABTECH) and is encouraged using AlphaScreen 680
Magnetic/570 emission filter group.IC50Calculate and use Prism 6.0.
2nd, cytoactive detection
MCF-7 and MDA-MB-231 cells are 5 × 104 cells/mls in the density of 96 hole flat-bottom microtiter plates.It is incubated
After 24 hours, the compound treatment of cell various concentrations.Cell survival experiment is determined using mtt assay.5mg/mL MTT are added
To in each hole.After being incubated 4 hours, solvent is removed, adding 150 microlitres of soluble crystal of DMSO.In 570nm mensuration absorbances
Value.
3rd, autophagy screening active ingredients
MCF-7 cells are incubated 6 hours with 5 μM of compound treatment.Then 0.05mM MDC are added to be incubated at 37 DEG C
30min.Using Flow Cytometry Assay cell fluorescence intensity.
The BRD4 inhibitory activity of 1 compound of table 1~42 and autophagy rate activity
Test result indicate that, compound of the invention has different degrees of inhibitory activity and autophagy activity to BRD4, its
In, compound 5, compound 15, compound 17, compound 18, compound 35, compound 36 effect preferably, optimal, chemical combination
Thing 36 (being designated as FL-411) has stronger inhibitory activity and autophagy activity to BRD4.
There is the related cell death of autophagy in the FL-411 inducing mammaries cancer cell of test example 2
It is observed that FL-411 can induce substantial amounts of cytoplasmic vacuoles in treatment MCF-7 and MDA-MB-231 cells
When, this is closely related with autophagy.Therefore, we examine whether FL-411 energy inducing mammary cancer cell generation autophagy.MDC is a kind of
The detection probe of autophagic vacuole, is applied to FL-411 treatment cells, is produced in fluorescence microscopy Microscopic observation and discovery of taking pictures a large amount of green
Color phosphor dot (Fig. 1).Then, we generate extensive vacuole (figure in the morphological landmarks of electric Microscopic observation autophagy in cytoplasm
2).Cell autophagy is confirmed (Fig. 3, Fig. 4) by the transfection of GFP-LC3 and LC3 immunofluorescences.In order to further confirm FL-411 energy
Enough induce autophagy, it has been found that FL-411 treated cell Beclin-1 expression increases, and autophagy substrate p62/SQSTM1 occurs
Lower；Autophagy mark LC3 II is presented the aggregation (Fig. 5) of time dependence simultaneously.Next, suppressing we used lysosome
Agent, BafA1 verifies the generation of autophagy stream.We have found that LC3-II and p62/SQSTM1 there occurs substantially in the presence of BafA1
Aggregation, show that autophagy stream is strengthened (Fig. 6) by FL-411.In order to the autophagy for further determining that FL-411 inductions is cytotoxicity
And cytoprotection, we apply autophagy inhibitor 3-MA, and it can suppress the formation of cell autophagy body by suppression
Type III phosphatidylinositol 3-kinase processed.Cell viability is have evaluated using mtt assay.We have found that cell survival rate is significantly increased to
After being processed with 3-MA (Fig. 7).Result above shows that FL-411 has arrived the related cell death of autophagy in MCF-7 and MDA-MB- again
In 231 cells.
The autophagy of the FL-411 of test example 3 inductions is by BRD4-AMPK-mTOR-ULK signal paths
Because BRD4 is the feedback of many tyrosine kinase activations, including PI3K/AKT.Therefore, believe we have studied autophagy
Number upstream mechanism come illustrate FL-411 induction autophagy.First, we have detected AKT, p-AKT (Ser473), p-Akt
(Thr308), the expression of mTOR and p-mTOR (ser2448) is after FL-411 treatment.As expected, FL-411 can be bright
It is aobvious to reduce p-Akt (Ser473), the expression of p-Akt (Thr308) and p-mTOR (ser2448), while the expression of Akt and mTOR
Constant (Fig. 8).Because ULK1 complexs are closely related with the formation of autophagosome.We have been then checked for ULK1, p-ulk1
(ser317) expression of, Atg13, p-atg13 (ser318) and FIP200.Result shows, after FL-411 treatment, ULK1 and
Atg13 phosphorylations are dramatically increased；And ULK1, Atg13 and FIP200 slightly have downward；Show that FL-411 can be by ULK1 complexs
Regulation autophagy (Fig. 9).It is special we used two kinds in order to whether the autophagy for exploring FL-411 inductions is only related to BRD4 suppression
Property siRNA suppress BRD4 expression.What is interesting is, it has been found that silence BRD4 can reduce the expression of c-myc proto-oncogenes, this
It is consistent with FL-411 treatment effects.But, different from the treatment of FL-411, the expression of AMPK α and ULK1 and phosphorylation and LC3-
II assembles and p62 degradeds can be changed after BRD4 strikes drop.This shows that the silence of BRD4 can not induce autophagy (Figure 10).It is comprehensive
On, the autophagy of FL-411 inductions is by BRD4-AMPK-mTOR-ULK signal paths.
The FL-411 of test example 4 has potential therapeutic action by targetting the autophagy that BRD4 regulates and controls in vivo
In order to evaluate antitumor activity in FL-411 bodies, two tumor of breast heteroplastic transplantation models are MCF-7 and MDA-MB-
231 cell models are used respectively.We used three kinds of FL-411 In vivo studies of various dose：25 mg/kgs, 50 millis
G kg and 100 mgs/kg.In all of model, FL-411 shows the inhibitory action of obvious tumour growth and is in
Dose dependent is respectively 80% and 76% (Figure 11, figure in the inhibition rate of tumor growth of MCF-7 and MDA-MB-231 cell models
12).It is observed that in all significant reduction (Figure 13) of tumor weight of all dosage groups.Also, FL-411 is displayed in all controlling
(Figure 14) is had no significant effect to body weight in treatment group.In order to the internal suppression tumour growth for checking whether FL-411 regulations is and subtracts
Few cell propagation is related to autophagy cell death is increased.We have carried out the SABC point of Ki-67 and LC3 to tumor tissues
Analysis.We have found that FL-411 can substantially reduce Ki-67 positive cell quantities and increase autophagy level (Figure 15).Next,
We conducted immunoblotting assay, mechanism in further clear and definite FL-411 bodies.We have found that FL-411 is capable of significantly LC3's II
Accumulation and caspase3 activate and suppress the expression of BRD4 and c-Myc.This is obtained with immunohistochemical analysis result and in vitro substantially
Result it is consistent (Figure 16).Additionally, we utilize zebra fish heteroplastic transplantation model, further confirm the antitumor work of FL-411
With.Compared with control group, it has been found that the FL-411 of all three dosage substantially suppresses the growth (Figure 17, Figure 18) of tumour.It is comprehensive
On, these results show that FL-411 has good antitumor activity in mouse xenografts and zebra fish model.
Claims (11)
1. compound or its pharmaceutically acceptable salt of the structural formula as shown in formula I：
Wherein, R1And R2It independently is hydrogen or C1-C4 alkyl, and R1And R2It is asynchronously hydrogen；Or R1And R2Connection forms ring-type
Structure, the cyclic structure is
R3It is phenyl ring or substituted phenyl ring；
N is 0 or 1.
2. compound according to claim 1 or its pharmaceutically acceptable salt, it is characterised in that：N is 0.
3. compound according to claim 2 or its pharmaceutically acceptable salt, it is characterised in that：R1And R2Connection is formed
It is preferred that R1And R2Connection is formedR3For
More preferably R1And R2Connection is formedR3For
4. compound according to claim 2 or its pharmaceutically acceptable salt, it is characterised in that：R1And R2It independently is
Hydrogen or C1-C4 alkyl, and R1And R2It is asynchronously hydrogen；
It is preferred that R1It is hydrogen；R2It is C1-C4 alkyl；R3For
More preferably R1It is hydrogen；R2ForR3For
More preferably R1It is hydrogen；R2ForR3For
5. compound according to claim 2 or its pharmaceutically acceptable salt, it is characterised in that：R1And R2Connection is formed
It is preferred that R1And R2Connection is formedR3For
6. compound according to claim 2 or its pharmaceutically acceptable salt, it is characterised in that：R1And R2Connection is formed
It is preferred that R1And R2Connection is formedR3For
More preferably R1And R2Connection is formedR3For
7. compound according to claim 2 or its pharmaceutically acceptable salt, it is characterised in that：R1And R2Connection is formed
It is preferred that R1And R2Connection is formedR3For
8. compound according to claim 1 or its pharmaceutically acceptable salt, it is characterised in that：N is 1；
It is preferred that n is 1, R1And R2Connection is formedR3For
More preferably n is 1, R1And R2Connection is formedR3For
9. compound according to claim 1 or its pharmaceutically acceptable salt, it is characterised in that：The type I compound
It is following compound：
10. compound or its pharmaceutically acceptable salt described in any one of claim 1~9 is preparing treatment breast cancer medicines
In purposes；It is preferred that the breast cancer treatment medicine is BRD4 inhibitor class medicines.
A kind of 11. pharmaceutical compositions for treating breast cancer, it is characterised in that：Claim of its active component comprising effective dose
Compound or its pharmaceutically acceptable salt described in 1~9 any one.
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| TAKASHI MASAOKA，等: "Exploiting Drug-Resistant Enzymes as Tools To Identify Thienopyrimidinone Inhibitors of Human Immunodeficiency Virus Reverse Transcriptase-Associated Ribonuclease H", 《J.MED.CHEM.》 * |
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