CN101537001A - Application of compound as JAK-STAT3 signal passage inhibitor - Google Patents

Abstract

The invention discloses an application of a compound as a JAK-STAT3 signal passage inhibitor, and particularly relates to an application of a compound with a formula I or pharmaceutically acceptable salt of the compound in the process of preparing an anti-tumor medicament. The application provides a novel treating candidate medicament for a tumor patient, thereby probably further improving the treatment effect on the patient and improving the prognosis of the patient. The compound with the formula I or the pharmaceutically acceptable salt of the compound has the effect on various tumor cells, which indicates that the compound can be used for treating various cancers including cerebral tumor, genitourinary system tumor, lymphatic system tumor, stomach cancer, laryngeal cancer, nasopharyngeal cancer, skin cancer, bone cancer, blood cancer, leukemia, breast cancer and histiocytic lymphoma, non-small cell lung cancer, small-cell lung cancer, lung adenocarcinoma, lung squamous cell cancer, pancreatic cancer, prostatic cancer, liver cancer, epithelial cell cancer and the like. The definition of genes in the formula refers to description for details.

Description

Application of compound as the JAK-STAT3 signal pathway inhibitor

Technical field

The present invention relates to the application of compound of a class as the JAK-STAT3 signal pathway inhibitor.

Technical background

Tumor is the No.1 killer of present human health and life, and its sickness rate is only second to cardiovascular disease.And along with the influence of environmental pollution and other factor, the sickness rate of malignant tumor is fast rise trend.According to the data that World Health Organization (WHO) 2003 announces, the total malignant tumor patient 1,000 ten thousand in the whole world in 2000, because of malignant tumor death person up to 6,200,000, account for 12%～25% of total death toll.Anticipate the year two thousand twenty, global annual new cases will reach 1,500 ten thousand.In recent years, though the exploitation listing of some novel targeting new drugs such as tyrosine protein inhibitor is arranged, but still can't satisfy growing clinical cancer patient's needs far away.The antitumor drug research and development also are still the important research direction of present medicament research and development circle.

Signal conduction and activating transcription factor (Signal Transducer and Activator of Transcription, STAT) be cytokine and growth factor receptors signal downstream effect device, usually the signal transduction that cell surface receptor can be produced is to nucleus, thereby and combines the expression of controlling gene with specific DNA promoter sequence in nuclear.There are seven different family members: Stat1, Stat2, Stat3, Stat5a, Stat5b, Stat6 in stat protein family.The STAT signal pathway is essential for normal cell physiological process, and fetal development for example, organ form and the performance function, congenital and day after tomorrow the acquired immunity function, and pair cell differentiation, growth and regulation of apoptosis.STAT3 is the family member of STATS, it in a large amount of tumor cell lines and human tumor by the unusual tyrosine kinase constitutively activate in upstream.Unusual STAT3 signal is by stimulating cellular proliferation, promote angiogenesis, suppresses apoptosis and participates in the generation and the development of human tumor.

The gene of coding STAT3 is positioned No. 17 chromosome (17q21.31) the mankind, and the STAT3 protein structure can be divided into the aminoacid conserved sequence, helical region, DNA of 6 functional areas: N end transcription activating district in conjunction with territory, bonding pad, SH2 domain and C end.Wherein the SH2 district is the most conservative and most important part of function in the STATs structure, participates in the tyrosine phosphorylation of stat protein, and plays an important role in the dimeric formation of STAT.Near 705 tyrosine phosphorylations this district are the activatory signs of STAT3.In addition, the C of stat protein end also has a serine phosphorylation site (Ser 727), is the substrate of multiple serine kinase, its phosphorylation level scalable STAT3 transcriptional activity [CHUNG J, UCHID A E, GRAMMERT C, et al.Mol Cell Biol, 1997 (17): 6508].STAT3 is bringing into play indispensable important function [CHAKRABORTY A, DYER K F, CASCIO M, et al.Blood, 1999,93 (1): 15] in the differentiation of fetal development and medullary cell in early days.In addition, STAT3 has also participated in active regulation and control [LEEMAN R J, LUI V W, GRANDIS J R.Expert Opin Biol Ther, 2006,6 (3): 231] such as cell growth, differentiation, vicious transformation and apoptosis inhibition.

The STAT3 signal pathway can pass through nonreceptor tyrosine kinase, growth factor receptors and three pathway activations of cytokine receptor.Nonreceptor tyrosine kinase activates participation [Smithgall T.E., Briggs S.D., Schreiner S., Lerner E.C., Cheng H., Wilson M.B.Oncogene, 19:2612-2618,2000 that STAT3 does not need receptor; Lin T.S., Mahajan S., Frank D.A.Oncogene, 19:2496-2504,2000; Danial N.N., Rothman P.Oncogene, 19:2523-2531,2000]; When cytokine such as IL-6 combine with the homoreceptor of cell surface, thereby the tyrosine kinase that the receptor dimerizationization activated receptor is correlated with is JAKs for example, and then kytoplasm section [the Darnell J.E. of activated receptor, Jr., Kerr I.M., Stark G.R.Science (Wash.DC), 264:1415-1421,1994; Ihle J.N.Curr.Opin.Cell Biol., 13:211-217,2001]; When somatomedin such as EGF and receptors bind because receptor autophosphorylation has tyrosine kinase activity in kytoplasm rear [Bromberg J., Darnell J.E., Jr.Oncogene, 19:2468-2473,2000 of receptor; Bowman T., Garcia R., Turkson J., Jove R.Oncogene, 19:2474-2488,2000].The receptor of tyrosine phosphorylation provides docking site to recruit the monomer STAT3[Heim M.H. in the kytoplasm, Kerr I.M., Stark G.R., Darnell J.E., Jr.Science (Wash.DC), 267:1347-1349,1995].Monomer STAT3 is by its SH2 domain and receptors bind and form dimer.[Darnell J.E.，Jr.Science(Wash.DC)，277：1630-1635，1997；Chen X.，Vinkemeier U.，Zhao Y.，Jeruzalmi D.，Darnell J.E.，Jr.，Kuriyan J.Cell，93：827-839，1998]。In time a few minutes, the transposition of STAT3 dimer is to nucleus, in nucleus with special DNA promoter sequence zygotic induction corresponding gene such as anti-apoptotic genes expression Bcl-XL, Mcl-1[Grandis J.R., Drenning S.D., Zeng Q., Watkins S.C., Melhem M.F., Endo S., Johnson D.E., Huang L., He Y., Kim J.D.Proc.Natl.Acad.Sci.USA, 97:4227-4232,2000; Epling-Burnette P.K., Liu J.H., Catlett-Falcone R., Turkson J., Oshiro M., Kothapalli R., Li Y., Wang J.M., Yang-Yen H.F., KarrasJ., Jove R., Loughran T.P., Jr.J.Clin.Investig., 107:351-362,2001; Epling-Burnette P.K., ZhongB., Bai F., Jiang K., Bailey R.D., Garcia R., Jove R., Djeu J.Y., Loughran T.P., Jr., Wei S.J.Immunol., 166:7486-7495,2001]; Cell cycle control gene c-Myc, cyclins D1/D2[Bowman T., BroomeM.A., Sinibaldi D., Wharton W., Pledger W.J., Sedivy J.M., Irby R., Yeatman T., Courtneidge S.A., Jove R.Proc.Natl.Acad.Sci.USA, 98:7319-7324,2001; Sinibaldi D., Wharton W., Turkson J., Bowman T., Pledger W.J., Jove R.Oncogene, 19:5419-5427,2000.]; Angiogenesis gene vegf expression [NIU G, WRIGHT K L, HUANG M, et al.Oncogene, 2002,21 (13): 2000-2008], thus pair cell propagation, survival, vascularization and tumor form to be regulated and control.

Under the normal physiological state, the activation of STAT3 is quick and of short duration, only keeps several minutes to several hours, plays key effect for Normocellular physiological function.STAT3 is the focus that converges of a plurality of carcinogenecity tyrosine kinase signal passages such as EGFR, IL-6/JAK, Src, is all finding to have persistence excessive activation (shown in chart 1) in the kinds of tumor cells.Induce unusual high expressed above-mentioned and cell proliferation, differentiation, the closely-related key gene of apoptosis behind the STAT3 excessive activation, promote cell proliferation, vicious transformation, block cell apoptosis by all means, show carcinogenesis.

The activation situation of table 1STAT in human tumor cell's (cell line or primary cell):

STAT3 is high expressed in the multiple malignant tumor of the mankind, and closely related with tumor differentiation degree, infiltration metastasis and prognosis.Therefore, the STAT3 micromolecule suppresses the new strategy that (JAK-STAT3 signal pathway inhibitor) can be used as clinical therapy of tumor.

At present the clinical treatment strategy at STAT3 mainly contains (1) STAT3 upstream tyrosine kinase inhibitor: 1. JAK family kinase inhibitors; 2. Src family kinase inhibitors; 3. EGF acceptor inhibitor; 4. Bcr-Abl tyrosine kinase inhibitor; (2) blocking-up STAT3 gene expression or protein function: 1. dominance-negative sense stat protein.STAT3 β is the clipped form of STAT3, still can form dimer and combines with DNA, but can not express by activated gene; 2. STAT3 antisense oligonucleotide (3) micromolecule suppresses STAT3 activation and dimerization.The report of several novel STAT3 micromolecular inhibitors such as at present existing Stattic and STA-21.

In recent years, along with the fast rise of new drug development cost, and the progressively downslide of drug development success rate, most of drug patent expires, and under the situation that the reserve new drug is difficult to follow up, drugmaker is the new patent protection medicine of exploitation on existing medicine basis one after another." old medicine is newly used " become a focus of International Pharmaceutical research and development.

Niclosamide (Niclosamide) has another name called niclosamide, Bayluscid, is synthesized successfully by German Shi Laofusi top grade people early than nineteen fifty-nine, carries out suitability for industrialized production by Bayer A.G.Originally be snail-killing medicine, has remarkable curative effect in schistosomicide treatment field, niclosamide can also be made Liniment with the prophylaxis of acute schistosomicide, kill oncomelania and schistosomulum, miracidium, have that sn ail control effect is good, toxic and side effects is little, to characteristics such as the toxicity of people and poultry are low, be the fresh combatants of treatment schistosomicide.In addition, niclosamide also is the people and the domestic animal expelling tenia medicine of a kind of wide spectrum, efficient, low toxicity.By income Chinese Pharmacopoeia, British Pharmacopoeia, European Pharmacopoeia etc.

This medicine is high for human its other homeothermia animal and plant safety, basic nonhazardous effect, and the human oral safe dose is up to more than the 2 gram/skies, and is only harmful to Fish.Clinical toxic and side effects is slight dizzy, uncomfortable in chest, abdominal discomfort or stomachache, heating, Pruritus etc.

At present, this medicine or derivatives thereof of Shang Weiyou or its analog and its esters are used for the reported in literature of cancer clinical treatment.

Summary of the invention

One of technical issues that need to address of the present invention provide the new application of a class as the chemical compound of JAK-STAT3 signal pathway inhibitor.

The technical scheme that solves the problems of the technologies described above is as follows:

Chemical compound or its pharmaceutically acceptable salt with formula I, as the application of JAK-STAT3 signal pathway inhibitor in the preparation antitumor drug,

Wherein:

Following a is 0 or 1; B is 0 or 1; N is optional to be 0,1,2 or 3;

Ar is not for containing or contain the aromatic rings of nitrogen, sulfur or oxygen heteroatom;

R ₁Certainly optional:

7)H；

8) halogen (F, Cl, Br, I);

9) O _aC ₁-C ₆Alkyl;

10) O _aC ₁-C ₆Cycloalkyl;

11) O _aC ₁-C ₆Heterocyclic radical;

12)O _aC ₀-C ₈-NR ₃R ₄；

Described alkyl, cycloalkyl and heterocyclic radical are optional to be selected from R by 0 or one or more ₃Substituent group replace;

R ₂Be 0～5 substituent group on the aromatic ring (Ar), each substituent group is certainly optional respectively:

14)H；

15) halogen;

16) (CO) _bO _aC ₁-C ₆Alkyl;

17) (CO) _bO _aC ₁-C ₆Cycloalkyl;

18) (CO) _bO _aC ₁-C ₆Heterocyclic radical;

19)(CO) _bO _aC ₀-C ₈-NR ₃R ₄；

20) C ₁-C ₃Perfluor or part fluoro-alkyl

21)(C＝O) _aNR ₆R ₅；

22)OH；

23)CN；

24)NO ₂；

25)SO ₂NR ₃R ₄；

26)COOH；

Described alkyl, cycloalkyl and heterocyclic radical are optional to be selected from R by 0 or one or more ₄Substituent group replace;

R ₃And R ₄Independently be selected from separately:

1)H；

2) (C=O) _aO _bC ₁-C ₆Alkyl;

3) (C=O) _aO _bAryl;

4) O _bC ₁-C ₃Perfluor or part fluoro-alkyl;

5) (C=O) _aO _bC ₃-C ₆Cycloalkyl;

6) (C=O) _aO _bHeterocyclic radical;

Described alkyl, aryl, cycloalkyl and heterocyclic radical are optional to be selected from R by 0～3 ₃Substituent group replace; Perhaps R ₃And R ₄And the atom that connects them forms the dicyclo that one 4～7 yuan monocycle or each ring are 4～7 yuan, and optionally contains 1～3 hetero atom that is selected from N, O and S, and described monocycle or dicyclo are optional by one or more R that are selected from ₃Substituent group replace.

Preferably, Ar is the chemical compound and the pharmaceutical salts thereof of phenyl; More preferably, Ar is a phenyl, and n is 0 chemical compound and pharmaceutical salts thereof; More preferably, niclosamide and pharmaceutical salts thereof are as the application of JAK-STAT3 signal pathway inhibitor in the preparation antitumor drug.

Preferably, described chemical compound has with one of Formula Il～IV structure:

R ₁～R ₄Definition same as described above.

The valid density of described chemical compound niclosamide or its pharmaceutically acceptable salt is 1 * 10 ^-8～1 * 10 ^-5M.

Another technical issues that need to address of the present invention provide a kind of antitumor Pharmaceutical composition.

Concrete technical scheme is as follows:

A kind of antitumor Pharmaceutical composition includes chemical compound or its pharmaceutically acceptable salt and pharmaceutically acceptable carrier with formula I structure.

Preferably, described tumor (cancer) is following any: histiocyte lymphatic cancer, nonsmall-cell lung cancer, small cell lung cancer, adenocarcinoma of lung, lung squamous cancer, cancer of pancreas, breast carcinoma, carcinoma of prostate, hepatocarcinoma, skin carcinoma, cell carcinoma, carcinoma of prostate, nasopharyngeal carcinoma, epidermal carcinoma, palace stem cancer, oral cancer, human fibrosarcoma, leukemia etc.

The invention provides chemical compound with formula 1 or its pharmaceutically acceptable salt purposes,, might further improve curative effect, improve its prognosis the patient for tumor patient provides new treatment drug candidate at the preparation antitumor drug.Chemical compound or its pharmaceutically acceptable salt with formula 1 can be used as AK-STAT3 signal path micromolecular inhibitor, the medicine that is used to prepare is effective to kinds of tumor cells, indicate this chemical compound have can be used for comprising cerebroma, urogenital system tumor, lymphsystem tumor, gastric cancer, laryngeal carcinoma, nasopharyngeal carcinoma, skin carcinoma, osteocarcinoma, leukemia, leukemia, breast carcinoma and histiocyte lymphatic cancer, nonsmall-cell lung cancer, small cell lung cancer, adenocarcinoma of lung, lung squamous cancer, cancer of pancreas, carcinoma of prostate, hepatocarcinoma, skin carcinoma, cell carcinoma, etc. multiple treatment for cancer.

In addition, described chemical compound can also be by increasing its clinical effectiveness with antitumor drug drug combination present application or that just locating the development phase.

Description of drawings

Fig. 1 is that B037 (niclosamide) is to the active sketch map of the inhibition of epidermal carcinoma;

Fig. 2 is that B037 (niclosamide) is to the active sketch map of the inhibition of breast carcinoma;

Fig. 3 is that B037 (niclosamide) is to the active sketch map of the inhibition of hepatocarcinoma;

Fig. 4 is the inhibition active sketch map of B037 (niclosamide) to palace stem cancer;

Fig. 5 is that B037 (niclosamide) is to the active sketch map of the inhibition of oral cancer;

Fig. 6 is that B037 (niclosamide) is to the active sketch map of human fibrosarcoma's inhibition;

Fig. 7 is that B037 (niclosamide) is to the active sketch map of the inhibition of colon cancer;

Fig. 8 is that B037 (niclosamide) is to the active sketch map of the inhibition of renal carcinoma;

Fig. 9 is that B037 (niclosamide) is to the active sketch map of the inhibition of carcinoma of prostate;

Figure 10 is that B037 (niclosamide) is to the active sketch map of the inhibition of pulmonary carcinoma;

Figure 11 is that western blot test shows that the B037 selectivity reduces the level of Cyclin D3, and the sketch map that the level of Cyclin E is not exerted an influence;

Figure 12 is that western blot test shows that the B037 selectivity suppresses the phosphorylation of STAT3, and the sketch map that the phosphorylation of MAPK signal path node albumen Raf, MEK, ErK is not exerted an influence;

Figure 13 is that the cut repairing test shows that B037 dose dependent ground suppresses the propagation of OS-RC-2 tumor cell and the sketch map of migration;

Figure 14 is that fluroimmunoassay shows that B037 dose dependent ground suppresses the sketch map of STAT3 to nuclear transfer.

The specific embodiment

Chemical compound of the present invention can have asymmetric center, chiral axis and chirality face, and racemate, racemate mixture and single diastereomer and all possible isomers and composition thereof of existing comprise that optical isomer includes in the present invention.In addition, chemical compound disclosed by the invention can tautomer exists, and two kinds of tautomeric forms all comprise within the scope of the invention, even only described wherein a kind of tautomeric structure.

As any variable (R for example ₁, R ₂, R ₃, R ₄Deng) in any component, occur surpassing once, then its each definition that occurs is independent of other each definition that occurs. and same, allow the combination of substituent group and variable, as long as this combination makes chemical compound stable.The line that puts loop systems from substituent group under represents that the key of indication can be connected on any annular atoms that can replace.If loop systems is. multi-ring, it means that this key only is connected on any suitable carbon atom of adjacent loops. be appreciated that those of ordinary skills can select the substituent group of The compounds of this invention and substitution pattern and provide chemically stable and can be easy to synthetic chemical compound from the raw material that can obtain easily by the method for art technology and following proposition.If being exceeded a group, substituent group self replaces, should understand these groups can be on the identical carbon atoms or on the different carbon atom, as long as make Stability Analysis of Structures. phrase " optional replaced " by one or more substituent groups be considered to phrase " optional replaced " by at least one substituent group quite and in the case embodiment preferred will have 0-3 substituent group.

Term used herein " alkyl " and " alkylidene " mean and comprise saturated fat alkyl side chain and straight chain with particular carbon atom number.For example, " C ₁-C ₆Alkyl " in " C ₁-C ₆" definition comprise the group of arranging with straight or branched with 1,2,3,4,5 or 6 carbon atom.For example, " C ₁-C ₆Alkyl " specifically comprises methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, the tert-butyl group, isobutyl group, amyl group, hexyl etc.Term " cycloalkyl " refers to have the monocycle saturated fat alkyl of particular carbon atom number.For example " cycloalkyl " comprises cyclopropyl, methyl-cyclopropyl, 2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopenta, cyclohexyl etc.

" alkoxyl " the cyclic or acyclic alkyl of the carbon atom that indicates number that connects by oxo bridge of representative.Therefore " alkoxyl " comprises the definition of abovementioned alkyl and cycloalkyl.

If the number of clear and definite carbon atom not, term " thiazolinyl " refers to straight chain, side chain or cyclic, contains the non-aromatic hydrocarbon base of 2-6 carbon atom and at least one carbon-to-carbon double bond.Carbon-to-carbon of preferred existence is two strong, and can have the most nearly 4 two being good for of nonaromatic carbon carbon.Therefore, " C ₂-C ₆Thiazolinyl " refer to have the thiazolinyl of 2-6 carbon atom.Thiazolinyl comprises vinyl, acrylic, cyclobutenyl, 2-methyl butene base and cyclohexenyl group.If the straight chain of thiazolinyl, side chain or annulus can contain two keys and indicate the thiazolinyl that replaces then this part can be substituted.

Term " alkynyl " refers to straight chain, a ripples or cyclic, contains 2-6 carbon atom and the triple-linked non-aromatic hydrocarbon base of at least one carbon carbon.Can have that the most nearly 3 carbon carbon three are strong.Therefore, " C ₂-C ₆Alkynyl " refer to have the alkynyl of 2-6 carbon atom.Alkynyl comprises acetenyl, propinyl, butynyl, 3-methyl butynyl etc.If the straight chain of alkynyl, side chain or annulus can contain triple bond and indicate the alkynyl that replaces then this part can be substituted.

In some example, substituent group can define with the certain limit carbon atom number that comprises 0, for example (C ₀-C ₆) alkylidene-aryl.If aryl is considered to phenyl, then this definition comprises phenyl self, also comprises-CH ₂Ph ,-CH ₂CH ₂Ph ,-CH (CH ₃) CH ₂CH (CH ₃) Ph etc.

The present invention's used " aryl " reaches in the finger ring in any stable monocycle of 7 atoms or each ring to reach 7 atom bicyclic carbocyclic, and wherein at least one ring is aromatic rings.The example of this aryl comprises phenyl, naphthyl, tetralyl, indanyl and xenyl.Aryl substituent be dicyclo and a ring be in the nonaromatic example, should understand through aromatic rings and connect.

7 atom bicyclic carbocyclic nearly in the stable monocycle of 7 atoms or each ring nearly in term used herein " heteroaryl " the representative ring, wherein at least one ring is for aromatic rings and contain 1-4 and be selected from O, the hetero atom of N and S. the heteroaryl in this range of definition includes but not limited to: bifurcation pyridine base, carbazyl, cinnolines base quinoxalinyl, pyrazolyl (pyrrazolyl), indyl, the benzotriazole base, the base of muttering of barking, thienyl, benzothienyl, the benzo base of muttering of barking, quinolyl, isoquinolyl oxazolyl isoxazolyl, indyl, pyrazinyl, pyridazinyl, pyridine radicals, pyrimidine radicals, pyrrole radicals, tetrahydroquinoline.For the definition of following heteroaryl, " heteroaryl " also is interpreted as and comprises any N-oxide derivative that contains the heteroaryl of nitrogen.The heteroaryl substituent group be dicyclo and to contain a ring be non-armaticity or do not contain in the heteroatomic example, should understand the aromatic rings or connect of respectively hanging oneself through containing heteroatomic ring.

Used term " heterocycle " or " heterocyclic radical " are meant and contain 1-4 heteroatomic 5 yuan of-10 yuan of armaticity or non-armaticity heterocycle that is selected from O, N and S among the present invention, and comprise bicyclic radicals. " heterocyclic radical " therefore comprises above mentioned heteroaryl, also comprises its dihydro and tetrahydro analog." heterocyclic radical " further example includes but not limited to: benzimidazolyl; benzofuranyl; benzopyranyl; the benzopyrazoles base; the benzotriazole base; benzothienyl benzoxazolyl; carbazyl; carbolinyl; the cinnolines base; furyl; imidazole radicals; indolinyl; indyl; indolazinyl; indazolyl; the different benzo base of muttering of barking; isoindolyl; isoquinolyl; isothiazolyl isoxazolyl; how pyridine radicals oxadiazole base oxazolyl oxazoline isoxazoline; oxetanyl (oxetanyl); pyranose; pyrazinyl; pyrazolyl; pyridazinyl; the pyridopyridine base; pyridazinyl; pyridine radicals; pyrimidine radicals; pyrrole radicals; quinazolyl; quinolyl quinoxalinyl; THP trtrahydropyranyl; tetrazole radical; the tetrazolo pyridine radicals; thiadiazolyl group; thiazolyl; thienyl; triazolyl; azetidinyl; 1; the 4-alkyl dioxin; azepan base (hexallydroazepinyl); piperazinyl; piperidyl; the pyridin-2-ones base; pyrrolidinyl; morpholinyl; thio-morpholinyl (thiomorpholinyl); the dihydrobenzo imidazole radicals; dihydro benzo furyl; the dihydrobenzo thienyl; Er hydrogen benzoxazolyl; the dihydrofuran base; the glyoxalidine base; indolinyl; the dihydro-isoxazole base; dihydro isothiazolyl Er Qing oxadiazole base dihydro-oxazole base; the dihydro pyrazinyl; the pyrazoline base; the dihydropyridine base; the dihydro-pyrimidin base; the pyrrolin base; the dihydroquinoline base; the dihydro tetrazole radical; the thiodiazoline base; dihydro-thiazolyl; the dihydro-thiophene base; the dihydro triazolyl; the dihydro azetidinyl; the methylenedioxyphenyl formoxyl; the tetrahydrochysene furan is fed base and tetrahydro-thienyl, and the N-oxide.The connection of heterocyclic substituent can realize by carbon atom or by hetero atom.

In one embodiment, heterocycle is selected from 2-azatropylidene (it is tall and erect down to go up grass) ketone, benzimidazolyl, 2 one diazepines (it is tall and erect down to go up grass) ketone, imidazole radicals, 2-imidazolone, indyl, isoquinolyl, morpholinyl, piperidyl, piperazinyl, pyridine radicals, pyrrolidinyl, 2-piperidones, 2-pyrimidone, 2-Pyrrolidone, quinolyl, tetrahydrofuran base, tetrahydro isoquinolyl and thienyl.

As understood by one of ordinary skill in the art, used among the present invention " halogen " (" halo ") or " halogen " mean and comprise chlorine, fluorine, bromine and iodine.

Unless otherwise defined, alkyl, thiazolinyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclic radical substituent group can be unsubstituted or replacement.For example, (C ₁-C ₆) alkyl can by one, two or three be selected from OH, oxo, halogen, alkoxyl, dialkyl amido or heterocyclic radical for example the substituent group of morpholinyl, piperidyl etc. replace.

In some example, definition R ₃And R ₄Make it form 4-7 unit monocycle jointly or each ring be the bicyclic heterocycle of 4-7 unit with the nitrogen that is connected them, and optional containing denitrogenate outer one or two and be selected from the other hetero atom of N, O and S, described heterocycle is chosen wantonly by one or more R that are selected from ⁵Substituent group replace.The heterocyclic example that can so form includes but not limited to following heterocycle, must keep that heterocycle is optional to be selected from R by one or more (and preferred one, two or three) firmly in mind ₃Substituent group replace:

R in one embodiment ₁Be selected from: hydrogen, halogen, hydroxyl, or (C ₁-C ₆) alkyl, alkoxyl.

In one embodiment, R ₂Be selected from and choose by hydrogen, halogen, alkyl, nitro, cyano group trifluoromethyl, amide groups, ester group, perfluor or part fluoro-alkyl, carboxyl, sulfoamido etc. wantonly.

In another embodiment, Ar is a substituted-phenyl; In another embodiment, Ar is for replacing pyrrole heavy stone used as an anchor base.

In another embodiment, n is 0.

In one embodiment, a is 0, and b is 1.Or in one embodiment, a is 0, and b is 0.

The present invention includes niclosamide and have the free form of the chemical derivative of formula I～IV structure, also comprise its pharmaceutically acceptable salt and stereoisomer.Some specific exemplary compounds are the protonated salt of aminated compounds among the present invention.Term " free form " refers to the aminated compounds with salt-independent shape.The pharmaceutically-acceptable salts that is included not only comprises the exemplary salt of specific compound of the present invention, also comprises the typical pharmaceutically acceptable salt of all formula I chemical compound free forms.Can use the free form of technical point known in the art from described chemical compound specific salts.For example, can be by for example NaOH dilute aqueous solution, potassium carbonate dilute aqueous solution, weak ammonia and sodium bicarbonate dilute aqueous solution are handled this salt and made free form regeneration with suitable alkali dilute aqueous solution.Free form some physical property for example in polar solvent on the dissolubility with its separately salt form more or less distinguish, but for the invention this hydrochlorate of purpose and alkali salt aspect other pharmacy with its free form is suitable separately.

Can be by the conventional chemical method from the synthetic pharmaceutically acceptable salt of the present invention of the The compounds of this invention that contains basic moiety or acidic moiety.Usually, the salt by ion exchange chromatography or the inorganic or organic acid prepared in reaction alkali compounds in the combination of appropriate solvent or multiple solvent by free alkali and stoichiometric amount or excessive required salt form.Similarly, by reacting the salt that forms acid compound with suitable inorganic or organic base.

Therefore, the pharmaceutically acceptable salt of The compounds of this invention comprises by alkaline The compounds of this invention and conventional the nontoxic salts inorganic or The compounds of this invention that organic acid reaction forms.For example, conventional nontoxic salts comprises and derives from for example salt of hydrochloric acid, hydrobromic acid, sulphuric acid, sulfamic acid, phosphoric acid, nitric acid etc. of mineral acid, also comprise from organic acid for example acetic acid, propanoic acid, succinic acid, glycolic, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pounce on the salt of preparations such as acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-anilinesulfonic acid., 2 one acetoxyl groups, one benzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane disulfonic acid, oxalic acid, hydroxyethylsulfonic acid., trifluoroacetic acid.

If The compounds of this invention is tart, then suitable " pharmaceutically acceptable salt " refers to comprise by pharmaceutically acceptable nontoxic alkali the salt of inorganic base and organic base preparation. the salt that derives from inorganic base comprises aluminum salt, ammonium salt, calcium salt, mantoquita, iron salt, ferrous salt, lithium salts, magnesium salt, manganese salt, manganous salt, potassium salt, sodium salt, zinc salt etc.Preferred especially ammonium salt, calcium salt, magnesium salt, potassium salt and sodium salt.Derive from the salt of pharmaceutically acceptable organic nontoxic alkali, described alkali comprises primary amine, the salt of secondary amine and tertiary amine, the amine that replaces comprises naturally occurring replacement amine, cyclic amine and deacidite be arginine for example, betanin, caffeine, choline, N, N '-dibenzyl-ethylenediamin, diethylamine, 2 one DEAE diethylaminoethanols, 2 one dimethylaminoethanols, ethylaminoethanol, ethanolamine, ethylenediamine, N one ethyl morpholine, N one ethyl piperidine, glucamine, glucosamine, histidine, hydroxocobalamin, isopropylamine, lysine, methyl glucoside amine, morpholine, piperazine, piperidines, the croak smack one's lips, the polyamines resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropyl amine (TPA), tromethane etc.

Because the acidic moiety of deprotonation is for example completed base and be can be anionic in the chemical compound under physiological condition, and this electric charge that has can be had cationic protonated or alkylating basic moiety by inside then, for example the former quantum balancing of quaternary nitrogen is offset, and is potential inner salt or amphion so should note The compounds of this invention.

The chemical compound that the present invention relates to have formula 1 that the present invention relates to (niclosamide and chemical derivative thereof), with and pharmaceutically acceptable salt as JAK-STAT3 signal path micromolecular inhibitor, and in the application of preparation in the antitumor drug.As skilled in the art to understand, related chemical compound and the acceptable salt of pharmacy thereof of the application is used for excess proliferative diseases such as the preparation treatment mankind and other mammiferous tumor.

In one embodiment, the application provides a kind of chemical derivative and excess proliferative disease or symptoms such as the acceptable salts for treating people of pharmacy or other mammal tumor of utilizing niclosamide and having formula I～IV structure.

In one embodiment, designed chemical compound and the acceptable salt of pharmacy thereof of the application can be used for the treatment of or control excess proliferative diseases such as histiocyte lymphatic cancer, nonsmall-cell lung cancer, small cell lung cancer, adenocarcinoma of lung, lung squamous cancer, cancer of pancreas, breast carcinoma, carcinoma of prostate, hepatocarcinoma, skin carcinoma, cell carcinoma, carcinoma of prostate, nasopharyngeal carcinoma, epidermal carcinoma, palace stem cancer, oral cancer, human fibrosarcoma, leukemia.

In one embodiment; the acceptable salt of chemical compound that the application is related and pharmacy thereof can with estrogenic agents present application or that just locating the development phase; androgen receptor modifier; the retinoid-like receptor modulators; cytotoxin/cytostatics; antiproliferative; protein transferase inhibitor; the HMG-CoA reductase inhibitor; the HIV kinases inhibitor; reverse transcriptase inhibitors; angiogenesis inhibitor; cell proliferation and the agent of existence signal suppressing; the medicine and the cell death inducer at the interference cell cycle outpost of the tax office; cytotoxic drug; the tyrosine protein inhibitor; the EGFR inhibitor; the VEGFR inhibitor; the serine/threonine protein inhibitor; the Bcr-Abl inhibitor; the c-Kit inhibitor; the Met inhibitor; the Raf inhibitor; mek inhibitor; the MMP inhibitor; topoisomerase enzyme inhibitor; the histidine deacetylase inhibitor; proteasome inhibitor; the CDK inhibitor; the Bcl-2 family protein inhibitor, the MDM2 family protein inhibitor; the IAP family protein inhibitor; the STAT family protein inhibitor; the PI3K inhibitor; the AKT inhibitor; the integrin blocker; interferon-' alpha '; il-1 2; cox 2 inhibitor; p53; the p53 activator; VEGF antibody; medication combined medications such as EGF antibody increase its clinical effectiveness.

The acceptable salt of chemical compound that the application is related and pharmacy thereof can be used for the treatment of following disease and following other disease of not listing according to following method:

1) a kind of utilization comprise the application related, utilize niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammiferous breast carcinoma.Including, but not limited to aggressive duct carcinoma, aggressive lobular carcinoma, DCIS and LCIS.

2) a kind of utilization comprise the application related, utilize niclosamide and have the chemical compound of chemical derivative and the Pharmaceutical composition treatment people of the acceptable salt of pharmacy or the method for other mammiferous respiratory cancer of formula I～IV structure.Including, but not limited to minicell, nonsmall-cell lung cancer and bronchial adenoma and pleura pulmonary blastoma.

3) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammiferous brain cancer.Including, but not limited to brain stem and glioma, cerebellum and big cerebral astrocytoma, ependymoma and neuroderm and strobile tumor body now.

4) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammiferous hero, female reproductive organ's tumor.The tumor of male reproductive organ includes but not limited to prostate and carcinoma of testis.Female reproductive organ's tumor includes but not limited to carcinoma of endometrium, cervical cancer, ovarian cancer, cancer of vagina and carcinoma vulvae and intrauterine tumor.

5) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammiferous gastral tumor.Include but not limited to anus cancer, colon cancer, colon straight way cancer, esophageal carcinoma, gastric cancer, cancer of pancreas rectal cancer, carcinoma of small intestine or glandula cancer.

6) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the tumor of the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammiferous urethra.Include but not limited to bladder cancer, carcinoma of penis, renal carcinoma, carcinoma of renal pelvis, carcinoma of ureter or carcinoma of urethra.

7) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammiferous cancer eye.Include but not limited to intraocular melanoma and retinocytoma.

8) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammal hepatocarcinoma.Include but not limited to hepatoma (having or do not have the stem cell cancer that fibre board changes), cancer of biliary duct (stones in intrahepatic bile duct cancer) and blended hepatocyte property cancer of biliary duct.

9) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammal skin cancer.Include but not limited to squamous cell carcinoma, Kaposi, malignant melanoma, Merck Schwann Cells skin carcinoma and non-melanoma cells cancer.

10) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammal head and neck cancer.Include but not limited to larynx, hypopharynx, nasopharynx, oropharynx cancer and lip and oral cancer.

11) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or the lymphadenomatous method of other mammal.Include but not limited to AIDS be correlated with lymphoma, non Hodgkin lymphoma, cutaneous T cell lymphoma, He Jiesen disease and central nervous system lymphoma.

12) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～V structure and the method for the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or other mammal sarcoma.Include but not limited to that soft tissue sarcoma, osteosarcoma, malignant fibrous histiocytoma, woods are sarcoma and rhabdomyosarcoma.

13) a kind of utilization comprise the application related, niclosamide and have the chemical derivative of formula I～IV structure and the Pharmaceutical composition of the acceptable salt of pharmacy treatment people or the leukemic method of other mammal.Include but not limited to acute myeloid leukemia, acute woods chronic myeloid leukemia, chronic lymphocyte leukemia, chronic lymphocytic leukemia and hairy cell leukemia.

Take mode and dosage range

According to the standard pharmaceutical technology, The compounds of this invention can give mammal, preferred people with pharmaceutically acceptable receptor, adjuvant or diluent combination separately or in Pharmaceutical composition.Can be oral or subcutaneous, intramuscular injection, intraperitoneal, vein, rectum and part, eyes, pulmonary, nasal cavity, parenteral give chemical compound.

In one embodiment, utilize niclosamide and have the chemical derivative treatment of formula I～IV structure or during patient such as control cancer, the taking dose scope is in oral 0.1～500 mg/day/kg body weight.Suitable administering mode is a single dose administration or every day secondary, three times, four inferior multiple dosings or utilize slow release method administration every day.For multiple large mammal, its preferred dosage scope is 0.1～1500 mg/day/kg body weight, is preferable over 0.5～100 mg/day/kg body weight.For average weight is 70 kilograms patient, and its, dosage was 1～500 milligram every day.For some property invigorated chemical compounds especially, adult patient's dosage every day can hang down and reach 0.1 mg/day.

Dosage form

This Pharmaceutical composition that contains active component can be made into and is suitable for the oral administration form, but for example tablet, buccal tablet, lozenge, water or oil suspension dispersion powder or granule, Emulsion, hard capsule or soft capsule or syrup or elixir.Can be according to the compositions of any known method preparation expection orally give in the Pharmaceutical composition manufacturing field, and for pharmaceutically purified and agreeable to the taste preparation is provided, this compositions can contain one or more medicaments that is selected from sweeting agent, flavoring agent, coloring agent and antiseptic.Tablet contains active component and the nontoxic acceptable accessories that is applicable to the manufacturing tablet.These adjuvants for example can be, and inert diluent is calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate for example; Granulation agent (granulating) and disintegrating agent be microcrystalline Cellulose, cross-linked carboxymethyl cellulose sodium (sodium crosscarmellose), corn starch or alginic acid for example; Binding agent is starch, gelatin, polyvinylpyrrolidone or arabic gum for example; Reach lubricant for example magnesium stearate, stearic acid or Pulvis Talci.Thereby tablet is coating or cover the disagreeable taste of medicine or prolong in gastrointestinal tract disintegrate and absorption and thereby provide the drug influence of last much longer by the known technology coating not.For example, the raw material that can adopt the water solublity taste masking is hydroxypropyl-methylcellulose or hyprolose for example, or adopts time-delay raw material for example ethyl cellulose, acetylbutyrylcellulose.Tabules can be 0.1 milligram/sheet, 0.2 milligram/sheet, 0.25 milligram/sheet, 0.5 milligram/sheet, and 1 milligram/sheet, 2 milligrams/sheet, 5 milligrams/sheet, 10 milligrams/sheet, 25 milligrams/sheet, 50 milligrams/sheet, 100 milligrams/sheet, 250 milligrams/sheet of and.Other dosage form such as capsule etc. can be done similar dosage reference.

The preparation that orally uses also can be made into hard-gelatin capsules, and wherein active component is mixed in inert solid diluent, for example in calcium carbonate, calcium phosphate or the kaolin; Or make Gelseal, wherein active component is mixed in water-solubility carrier for example in macrogol or oil medium such as Oleum Arachidis hypogaeae semen, liquid paraffin or the olive oil.

Aqueous suspension contains and the blended active material of adjuvant that is suitable for making aqueous suspension.This adjuvant is for example sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl emthylcellulose, sodium alginate, polyvinylpyrrolidone, tragakanta and an arabic gum of suspending agent; Dispersant or wetting agent can be for example lecithin of naturally occurring phospholipid, or the condensation product of alkylene oxide and fatty acid Myrj 45 for example, or the condensation product of alkylene oxide and long-chain fatty alcohol 17 carbon ethyleneoxy hexadecanol (heptadecaethyleneoxycetanol) for example, or alkylene oxide and derive from fatty acid and the condensation product of the partial ester of hexitol polyoxyethylene sorbitol monooleate for example, or alkylene oxide and derive from fatty acid and the condensation product of the partial ester of hexitan polyethylene dehydrated sorbitol mono-fatty acid ester for example.This aqueous suspension also can contain one or more antiseptic for example ethylparaben or P-hydroxybenzoic acid n-pro-pyl ester, one or more coloring agent, and one or more flavoring agents and one or more sweeting agents be sucrose, glucide or aspartame for example.

Can be by active component being suspended in vegetable oil for example in Oleum Arachidis hypogaeae semen, olive oil, Oleum Sesami or the Oleum Cocois, or mineral oil for example prepares oil-based suspension in the liquid paraffin.This oil-based suspension can contain thickening agent for example Cera Flava, hard paraffin or spermol.Can add sweeting agent mentioned above and flavoring agent and be fit to oral preparation to provide.Can for example butylated hydroxyanisole (butylated hydroxyanisol) or alpha tocopherol store these compositionss by adding antioxidant.

But dispersion powder or granule are suitable for preparing aqueous suspension and providing and dispersant or wetting agent, suspending agent and the blended active component of one or more antiseptic by adding entry.Suitable dispersant or wetting agent and suspending agent be the example explanation by above relating to.Also can there be other adjuvants for example sweeting agent, flavoring agent and coloring agent.These compositionss can by add antioxidant for example ascorbic acid store.

The present composition also can be made into the form of oil-in-water emulsion.Oil phase can be vegetable oil for example olive oil or Oleum Arachidis hypogaeae semen, or mineral oil for example liquid paraffin or its mixture.Suitable emulsifying agent can be for example soybean lecithin of naturally occurring phospholipid, reach esters or derive from fatty acid and the partial ester of hexitan mixture, the condensation product of for example sorbitan monooleate, and described partial ester and alkylene oxide is the polyoxyethylene sorbitan monooleate for example.This Emulsion also can contain sweeting agent, flavoring agent, antiseptic and antioxidant.

For example glycerol, propylene glycol, sorbitol or sucrose prepare syrup and elixir can to use sweeting agent.This preparation also can contain wetting agent, antiseptic, flavoring agent and coloring agent and antioxidant.

Pharmaceutical composition can be made into the aqueous solution of sterile injectable.In acceptable carrier and solvent, can adopt water, ringer's solution and isotonic sodium chlorrde solution.

This sterile injectable preparation also can be made into active component and is dissolved in sterile injectable oil-in-water microemulsion in the oil phase.For example, at first active component is dissolved in the mixture of Oleum Glycines and lecithin, then oil solution is put into the mixture of water and glycerol and handled and make microemulsion.

This injectable solution or microemulsion can pass through local bolus injection (local bolus injection) and import in patient's blood flow.Selectable, in this way give the constant circulation concentration that solution or microemulsion help keeping chemical compound.For keeping this constant density, can utilize the continuous intravenous injection delivery apparatus.An embodiment of this device is DeltecCADD-PLUS ^TMModel 5400 intravenous injection pumps.

This Pharmaceutical composition can be made into sterile injectable solution or the oily suspension form that is used for intramuscular or subcutaneous administration.Can use the dispersant above mention or wetting agent and suspending agent to prepare this suspension according to known technology.Sterile injectable preparation also can be made into sterile injectable solution or the suspension in nontoxic parenteral acceptable diluent or solvent, for example as the solution in 1,3 butylene glycol.In addition, the conventional fixed oil that adopts is as solvent or suspension medium.For this purpose, can adopt any non-irritating fixed oil to comprise synthetic monoglyceride or diglyceride.In addition, find in injectable formulation, to use for example oleic acid of fatty acid.

Niclosamide and chemical derivative with formula I～IV structure thereof also can rectally the suppository form administration.Can prepare these compositionss by hybrid medicine and suitable nonirritant adjuvant, thus its be at normal temperatures solid but under rectal temperature for liquid and therefore in rectum fusing discharge medicine.This raw material comprises the mixture and the cithrol of cocoa butter, glycerin gelatine, hydrogenant vegetable oil, various molecular weight polyisoprene ethylene glycol.

Use for the part, adopt emulsifiable paste, ointment, gel, solution or the suspension etc. (be this application purpose, topical application comprises mouth wass and collutory) that contain niclosamide and have the chemical derivative of formula I～IV structure.

The compounds of this invention can use with the intranasal form administration through the part of suitable intranasal carrier and delivery apparatus, or uses the form administration of the skin patch that those of ordinary skills know through skin.After the administration of transdermal delivery system form, the dosage of whole dosage regimen is naturally than intermittently administration is continuous.The compounds of this invention also can be sent by suppository, and the substrate of employing is the mixture and the cithrol of cocoa butter, glycerin gelatine, hydrogenant vegetable oil, various molecular weight polyisoprene ethylene glycol for example.

If give the experimenter The compounds of this invention, normally by the doctor that prescribes usually according to each patient's age, body weight, sex and reaction, and the corresponding change dosage of patient's serious symptom and determine daily dose.

Drug metabolite and prodrug

The niclosamide that the application is related and have the chemical derivative of formula I～IV structure and the metabolite of the acceptable salt of pharmacy, and the prodrug that can change the structure of related chemical compound of the application and the acceptable salt of pharmacy thereof in vivo into, be also contained in the application's the claim.

Drug combination

Niclosamide and the chemical derivative with formula I～IV structure thereof can or improve the other medicines coupling of similar condition of illness to known treatment.During administering drug combinations, the original Gei Yaofangshi ﹠amp of medicine; Dosage remains unchanged, and simultaneously or the chemical derivative of taking niclosamide subsequently and having formula I～IV structure.When niclosamide and have the chemical derivative of formula I～IV structure and other one or more medicines when taking simultaneously, preferred use contains one or more known drugs and niclosamide simultaneously and has the Pharmaceutical composition of the chemical derivative of formula I～IV structure.Drug combination is also included within chemical derivative and other one or more known drugs that the eclipsed time period takes niclosamide and have formula I～IV structure.When niclosamide and one or more medicines of chemical derivative and other with formula I～IV structure thereof carry out drug combination niclosamide and have the formula I～chemical derivative of IV structure or the dosage of the dosage of known drug may be than their independent medications the time lower.

Can carry out the medicine of drug combination or active component with niclosamide and the chemical derivative with formula I～IV structure thereof comprises but is not limited to:

Estrogenic agents; androgen receptor modifier; the retinoid-like receptor modulators; cytotoxin/cytostatics; antiproliferative; protein transferase inhibitor; the HMG-CoA reductase inhibitor; the HIV kinases inhibitor; reverse transcriptase inhibitors; angiogenesis inhibitor; cell proliferation and the agent of existence signal suppressing; the medicine and the cell death inducer at the interference cell cycle outpost of the tax office; cytotoxic drug; the tyrosine protein inhibitor; the EGFR inhibitor; the VEGFR inhibitor; the serine/threonine protein inhibitor; the Bcr-Abl inhibitor; the c-Kit inhibitor; the Met inhibitor; the Raf inhibitor; mek inhibitor; the MMP inhibitor; topoisomerase enzyme inhibitor; the histidine deacetylase inhibitor; proteasome inhibitor; the CDK inhibitor; the Bcl-2 family protein inhibitor, the MDM2 family protein inhibitor; the IAP family protein inhibitor; the STAT family protein inhibitor; the PI3K inhibitor; the AKT inhibitor; the integrin blocker; interferon-' alpha '; il-1 2; cox 2 inhibitor; p53; the p53 activator; VEGF antibody; EGF antibody etc.

In one embodiment, can carry out the medicine of drug combination or active component with niclosamide and the chemical derivative with formula I～V structure thereof comprises but is not limited to: aldesleukin, alendronic Acid, interferon, Ah Qu Nuoying, allopurinol, allopurinol sodium, the palonosetron hydrochlorate, altretamine, amino glutethimide, amifostine, amrubicin, SN-11841, arimidex, dolasetron, aranesp, arglabin, arsenic trioxide, Arnold is new, U-18496, azathioprine, bacillus calmette-guerin vaccine or tice bacillus calmette-guerin vaccine, bestatin, betamethasone acetate, betamethasone sodium phosphate preparation, bexarotene, Bleomycin Sulphate, broxuridine, bortezomib, busulfan, calcitonin, A Laizuo monoclonal antibody injection, capecitabine, carboplatin, Kang Shide, cefesone, celmoleukin, daunorubicin, chlorambucil, cisplatin, cladribine, cladribine, chlorine is bent phosphoric acid, cyclophosphamide, arabinose born of the same parents former times, dacarbazine, actinomycin D, daunorubicin liposome, dexamethasone, dexamethasone phosphate, estradiol valerate, denileukin diftitox, Di Bomei, deslorelin, the dilazep assistant is given birth to, diethylstilbestrol, Fluconazole, many Xi Taqi, doxifluridine, amycin, dronabinol, admire-the 166-chitosan complexes, eligard, rasburicase, epirubicin hydrochloride, aprepitant, epirubicin, Epoetin Alfa, erythropoietin, according to platinum, Ergamisole, estradiol preparation, 17-, estramustine phosphate sodium, ethinylestradiol, amifostine, hydroxyl phosphoric acid, Etopophos, etoposide, fadrozole, the tamoxifen preparation, filgrastim, Fei Nasi carries, Fei Leisi replaces, floxuridine, fluconazol, fludarabine, the 5-fluorodeoxyuridine monophosphate, 5-fluorouracil, fluoxymesterone, flutamide, formestane, 1-β-D-arabinofuranosylcytosine-5 '-stearoyl phosphate ester, fotemustine, fulvestrant, gamma globulin, gemcitabine, gemtuzumab Ozogamicin Mylotarg CDP 771, imatinib mesylate, Gliadel, goserelin, Graniseeron Hydrochloride, histrelin, new with U.S., hydrocortisone, erythro-hydroxyl nonyl adenine, hydroxyurea, replace not monoclonal antibody of smooth different shellfish, idarubicin, ifosfamide, interferon-ALPHA, interferon-' alpha ' 2, interferon A, interferon B, interferon alfa-n1, Alferon N, interferon beta, interferon gamma-1a, interleukin II, intron A, Iressa, Irinotecan, Kytril, the sulphuric acid lentinan, letrozole, formyl tetrahydrofolic acid, leuprorelin, leuprolide acetate, L-tetramisole, the levo leucovorin calcium salt, levothyroxine sodium, levothyroxine sodium preparation, lomustine, lonidamine, dronabinol, chlormethine, mecobalamin, medroxyprogesterone acetate, megestrol acetate, melphalan, esterified estriol, 6-coloured glaze base purine, mesna, methotrexate, the amino-laevulic acid methyl ester, miltefosine, minocycline, ametycin, mitotane, rice holder Herba Alii fistulosi quinone, trilostane, the citric acid Evacet, nedaplatin, the Pegylation filgrastim, oprelvekin, neupogen, nilutamide, tamoxifen, NSC-631570, recombination human interleukins-11-β, octreotide, Ondansetron Hydrochloride, the dehydrohydro-cortisone oral solution, oxaliplatin, paclitaxel, Pediapred, pegaspargase, Pai Luoxin, pentostatin, Picibanil, the hydrochloric acid pilocarpine, adjoin the gentle star that compares, plicamycin, porfimer sodium, prednimustine, prednisolone steaglate, prednisone, premarin, the third kappa umbilicus, the epoetin, Raltitrexed, Libiee, etidronic acid rhenium-186, Mabthera, Redoxon-A, romurtide, Salagen, octreotide, sargramostim, semustine, sizofiran, sobuzoxane, bluff the sodium prednisolone, Paphos acid, stem-cell therapy, streptozocin, Metastron, levothyroxine sodium, tamoxifen, YM-617, Ta Suonaming, tastolactone, taxotere, teceleukin, the temozolomide, teniposide, Testosterone Propionate, methyltestosterone, thioguanine, thio-tepa, thyrotropin, tiludronic acid, topotecan, toremifene, tositumomab, Herceptin, treosulfan, tretinoin, the methotrexate tablet, the trimethyl melamine, trimetrexate, the acetic acid triptorelin, triptorelin pamoate, UFT, uridnine, valrubicin, vesnarinone, vinblastine, vincristine, vindesine, vinorelbine, virulizin, ADR-529, Zinostatin stimalamer, ondansetron, the paclitaxel protein stabilization formulations, acolbifene, interferon r-lb, affinitak, aminopterin-induced syndrome, arzoxifene, asoprisnil, atamestane, atrasentan, BAY43-9006, Avastin, CCI-779, CDC-501, celecoxib, Cetuximab, crisnatol, cyproterone acetate, decitabine, DN-101, amycin-MTC, dSLIM, the dutasteride, edotecarin, eflornithine, Exatecan, fenretinide, histamine dihydrochloric acid, the histrelin hydrogel implant, holmium-166 DOTMP, ibandronic acid, interferon gamma, intron-PEG, ixabepilone, keyhole shape hemocyanin, L-651582, lanreotide, lasofoxifene, libra, lonafamib, Miproxifene, minot is bent acid esters, MS-209, liposome MTP-PE, MX-6, nafarelin, Nemorubicin, Neovastat, Nola Qu Te, Ao Limosen, onco-TCS, osidem, paclitaxel polyglutamic acid esters, pamidronate disodium injection, PN-401, QS-21, overstate the West, R-1549, raloxifene, ranpirnase, 13-is along tretinoin, husky platinum, seocalcitol, T-138067, tarceva, the docosahexenoic acid paclitaxel, thymosin, loud, high-pitched sound azoles furan woods, tipifarnib, tirapazamine, TLK-286, toremifene, trans MID-lo7R, valspodar, vapreotide, vatalanib, Verteporfin, vinflunine, Z-100 and azoles come unicorn acid or their combination.

The purpose of the embodiment that is provided is to help further to understand the present invention.Adopting specific raw material, species is for describing the present invention rather than limiting its reasonable range.

Embodiment 1

5-chloro-N-(4-fluorophenyl)-2-hydroxybenzamide

(5-chloro-N-(4-fluorophenyl)-2-hydroxybenzamide)

Synthetic method:

Mix 1mmol 5-chloro-2-methoxybenzoic acid and 2ml thionyl chloride, reflux one hour.Steam and remove thionyl chloride, add anhydrous tetrahydro furan 2ml dilution then, stand-by.

The 4-fluoroaniline compounds of 2mmol is dissolved in the 2ml anhydrous tetrahydro furan, is added drop-wise in the 5-chloro-2-methoxy benzoyl chloride solution for preparing previously stirred overnight at room temperature then at ambient temperature.After the thin layer chromatography detection reaction finishes, under the room temperature, add suitable quantity of water and dichloromethane, mixture changes separatory funnel over to, gets organic layer.Organic layer solution is used an amount of diluted sodium hydroxide solution (1 time) successively, saturated nacl aqueous solution (1 time), 5% hydrochloric acid solution (2 times), saturated nacl aqueous solution (1 time) washing, use anhydrous sodium sulfate drying then, remove solvent under reduced pressure and obtain solid, need not be further purified.

The solid of preparation all is dissolved in the 5ml anhydrous methylene chloride stirring at room, argon shield.Boron tribromide 0.1ml slowly drips in the foregoing solution with the dilution of 2ml anhydrous methylene chloride, stirring at room, and the thin layer chromatography monitoring reaction, reaction finishes.Slowly add 4M potassium hydroxide solution 2ml, add suitable quantity of water and dichloromethane again, change separatory funnel then over to, the water intaking layer.Concentrated hydrochloric acid is added dropwise to the water layer in the stirring, regulates pH to separating out a large amount of solids, then with the acetic acid ethyl dissolution extraction, and the washing of organic layer reuse saturated sodium-chloride, anhydrous sodium sulfate drying.Remove solvent under reduced pressure, obtain product, need not be further purified.

¹HNMR(400MHz，DMSO-d ₆)，δ12.12(s，1H)，9.92(s，1H)，8.02(d，J＝1.6Hz，1H)，7.77～7.80(m，3H)，7.46～7.49(m，1H)，7.15～7.20(m，3H)，7.00(d，J＝8.8Hz，1H)；

MS(ES+APCI)，m/z：266.1(M+H) ⁺，264.0(M-H) ^-

Embodiment 2

5-chloro-N-(3, the 4-difluorophenyl)-2-hydroxybenzamide

(5-chloro-N-(3，4-difluorophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.60(s，1H)，10.52(s，1H)，7.86～7.91(m，2H)，7.43～7.48(m，3H)，7.46～7.49(m，1H)，7.02(d，J＝1.6Hz，1H)

MS(ES+APCI)，m/z：284.1(M+H) ⁺，282.0(M-H) ^-

Embodiment 3

5-chloro-N-(4-(4-morphine quinoline base) phenyl)-2-hydroxybenzamide

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ10.41(s，1H)，7.99(d，J＝2.0Hz，1H)，7.54(d，J＝8.8Hz，2H)，7.43～7.46(m，1H)，6.94～6.99(m，3H)，

MS(ES+APCI)，m/z：333.0(M+H) ⁺，331.2(M-H) ^-

Embodiment 4

5-chloro-N-(4-nitrile phenyl)-2-hydroxybenzamide

(5-chloro-N-(4-cyanophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.48(s，1H)，10.66(s，1H)，7.89(d，J＝8.4Hz，2H)，7.80～7.82(m，2H)，7.44～7.47(m，1H)，7.00(d，J＝8.8Hz，1H)

MS(APCI)，m/z：279.1(M+H) ⁺，271.0(M-H) ^-

Embodiment 5

5-chloro-N-(4-trifluoromethyl)-2-hydroxybenzamide

(5-chloro-2-hydroxy-N-(4-(trifluoromethyl)phenyl)benzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ10.66(s，1H)，7.93(d，J＝7.6Hz 2H)，7.88(d，J＝2.8Hz，1H)，7.74(d，J＝8Hz，2H)，7.46～7.49(m，1H)，7.03(d，J＝8.8Hz，1H)

MS(APCI)，m/z：316.0(M+H) ⁺，314.0(M-H) ^-

Embodiment 6

5-chloro-N-(3.5-difluorophenyl)-2-hydroxybenzamide

(5-chloro-N-(3，5-difluorophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ10.61(s，1H)，7.81(d，J＝2.4Hz 1H)，7.46～7.50(m，3H)，6.98～7.04(m，2H)，

MS(APCI)，m/z：284.0(M+H) ⁺，282.0(M-H) ^-

Embodiment 7

5-chloro-N-(3-chloro-4-fluorophenyl)-2-hydroxybenzamide

(5-chloro-N-(3-chloro-4-fluorophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.60(s，1H)，10.50(s，1H)，8.01～8.03(m，1H)，7.87(s，1H)，7.64～7.66(m，1H)，7.41～7.48(m，2H)，7.02(d，J＝8.8Hz 1H)

MS(APCI)，m/z：300.0(M+H) ⁺，298.0(M-H) ^-

Embodiment 8

5-chloro-N-(3-acetylene phenyl)-2-hydroxybenzamide

(5-chloro-N-(3-ethynylphenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.69(s，1H)，10.45(s，1H)，7.91(d，J＝9.2Hz，1H)，7.72(d，J＝8.0Hz 1H)，7.49(d，J＝2.4Hz 1H)，7.38～7.42(m，1H)，7.26(d，J＝7.6Hz 1H)，7.03(d，J＝8.8H，1H)，4.22(s，1H)

MS(APCI)，m/z：272.0(M+H) ⁺，270.0(M-H) ^-

Embodiment 9

5-chloro-N-(3,4, the 5-trifluorophenyl)-2-hydroxybenzamide

(5-chloro-2-hydroxy-N-(3，4，5-trifluorophenyl)benzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d6)，δ11.47(s，1H)，10.63(s，1H)，7.82(s，1H)，7.68～7.72(m，2H)，7.47～7.49(m，1H)，7.02～7.07(m，1H)

MS(APCI)，m/z：302.1(M+H) ⁺，300.0(M-H) ^-

Embodiment 10

5-chloro-2-hydroxy-n-(3,4, the 5-trimethylphenyl) Benzoylamide

(5-chloro-2-hydroxy-N-mesitylbenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ12.36(s，1H)，10.03(s，1H)，8.06～8.09(mt，1H)，7.49～7.51(m，1H)，7.01(d，J＝8.8Hz 1H)，6.95(s，2H)，2.26(s，3H)，2.14(s，6H)

MS(APCI)，m/z：290.0(M+H) ⁺，288.1(M-H) ^-

Embodiment 11

5-chloro-N-(3-fluorophenyl)-2-hydroxybenzamide

(5-chloro-N-(3-fluorophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.62(s，1H)，10.51(s，1H)，7.87(s，1H)，7.69(d，J＝11.6Hz，1H)，7.45(d，J＝7.2Hz，2H)，7.37～7.41(t，1H)，7.00(d，J＝8.4Hz，1H)，6.93～6.95(m，1H)

MS(APCI)，m/z：266.0(M+H) ⁺，264.0(M-H) ^-

Embodiment 12

5-chloro-N-(2,3, the 4-trifluorophenyl)-2-hydroxybenzamide

(5-chloro-2-hydroxy-N-(2，3，4-trifluorophenyl)benzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ12.13(s，1H)，10.67(s，1H)，7.92(s，1H)，7.88(d，J＝5.6Hz，1H)，7.48(d，J＝8.8Hz 1H)，7.33～7.38(m，1H)，7.03(d，J＝8.8Hz，1H).

MS(APCI)，m/z：300.0(M+H) ⁺，302.1.0(M-H) ^-

Embodiment 13

5-chloro-N-(4-bromophenyl)-2-hydroxybenzamide

(N-(4-bromophenyl)-5-chloro-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ10.48(s，1H)，7.91(d，J＝2.4Hz，1H)，7.69(d，J＝8.8Hz，2H)，7.55(d，J＝8.8Hz，2H)，7.44～7.47(m，1H)，7.01(d，J＝8.8Hz，1H)

MS(APCI)，m/z：326.0(M+H) ⁺，324.0(M-H) ^-

Embodiment 14

5-chloro-N-(4-aminomethyl phenyl)-2-hydroxybenzamide

(N-(4-bromophenyl)-5-chloro-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.98(s，1H)，10.35(s，1H)，7.98(d，J＝2.4Hz，1H)，7.57(d，J＝8.0Hz，2H)，7.45～7.47(m，1H)，7.18(d，J＝8.0Hz，2H)

7.01(d，J＝8.8Hz，1H)

MS(APCI)，m/z：262.0(M+H) ⁺，260.1(M-H) ^-

Embodiment 15

5-chloro-N-(4-nitrogen pyridine radicals)-2-hydroxybenzamide

(5-chloro-2-hydroxy-N-(pyridin-4-yl)benzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.25(s，1H)，8.61，(s，2H)，8.00，(s，2H)，7.83，(s，1H)，7.46，(d，J＝8.4Hz，1H)，7.22(d，J＝8.4Hz 1H)

MS(APCI)，m/z：249.0(M+H) ⁺，247.0(M-H) ^-

Embodiment 16

5-chloro-N-(2-chloro-4-trifluoromethyl)-2-hydroxybenzamide

(5-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ12.40(s，1H)，11.14(s，1H)，，8.71(d，J＝8.8Hz，1H)，7.95(d，J＝2.8Hz，2H)，7.72～7.76(m，1H)，7.49～7.54(m，1H)，7.08(d，J＝8.8Hz，1H)

MS(APCI)，m/z：349.9(M+H) ⁺，348.0(M-H) ^-

Embodiment 17

5-chloro-N-(4-chlorphenyl)-2-hydroxybenzamide

(5-chloro-N-(4-chlorophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d6)，δ11.67(s，1H)，10.45(s，1H)，7.90(s，1H)，7.73(d，J＝8.4Hz 2H)，7.40～7.46(t，2H)，7.00(d，J＝8.8Hz，1H).

Embodiment 18

5-chloro-N-(2, the 4-Dichlorobenzene base)-2-hydroxybenzamide

(5-chloro-N-(2，4-dichlorophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d6)，δ12.29(s，1H)，10.92(s，1H)，8.44(d，J＝9.2Hz，1H)，7.98(d，J＝2.4Hz，1H)，7.75(d，J＝1.6Hz，1H)，7.48～7.51(m，2H)，7.08(d，J＝8.4Hz 1H).

MS(ES+APCI)，m/z：315.9(M+H) ⁺，313.9(M-H) ^-

Embodiment 19

5-chloro-N-(3-chlorphenyl)-2-hydroxybenzamide

(5-chloro-N-(3-chlorophenyl)-2-hydroxy

benzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d6)，δ11.61(s，1H)，10.50(s，1H)，7.89～7.92(m，2H)，7.61～7.63(m，1H)，7.46～7.49(m，1H)，7.39～7.43(m，1H)，7.20～7.22(m，1H)，7.03(d，J＝8.8Hz，1H)

MS(ES+APCI)，m/z：282.0(M+H) ⁺，280.0(M-H) ^-

Embodiment 20

5-chloro-N-(4-methoxyphenyl)-2-hydroxybenzamide

5-(chloro-2-hydroxy-N-(4-methoxyphenyl)benzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ12.08(s，1H)，10.32(s，1H)，8.01(d，J＝2.8Hz，1H)，7.60(d，J＝8.8Hz，2H)，7.45～7.48(m，1H)，7.00(d，J＝8.8Hz，1H)，6.96(d，J＝8.8Hz，2H)，3.76(s，3H)

MS(ES+APCI)，m/z：278.0(M+H) ⁺，276.0(M-H) ^-

Embodiment 21

5-chloro-N-(3-methoxyphenyl)-2-hydroxybenzamide

(5-chloro-2-hydroxy-N-(3-methoxyphenyl)benzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.81(s，1H)，10.38(s，1H)，7.95(d，J＝2.4Hz，1H)，7.46～7.47(m，1H)，7.40(s，1H)，7.25～7.31(m，2H)，7.01～7.06(t，1H)，6.72～6.75(m，1H)，3.77(s，3H)

MS(ES+APCI)，m/z：278.0(M+H) ⁺，276.0(M-H) ^-

Embodiment 22

5-chloro-N-benzyl-2-hydroxybenzamide

(N-benzyl-5-chloro-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ12.50(s，1H)，9.37～9.40(t，1H)，7.99～8.00(d，J＝2.4Hz，1H)，7.44～7.47(m，1H)，7.35(d，J＝4.4Hz，4H)，7.26～7.29(m，1H)，6.96(d，J＝8.8Hz 1H))，4.52(d，J＝5.6Hz，2H)

MS(ES+APCI)，m/z：262.0(M+H) ⁺，260.0(M-H) ^-

Embodiment 23

5-chloro-N-phenyl-2-hydroxybenzamide

(5-chloro-2-hydroxy-N-phenylbenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d6)，δ11.86(s，1H)，10.40(s，1H)，7.98(s，1H)，7.70(d，J＝7.6Hz，2H)，7.47～7.49(m，1H)，7.36～7.40(m，2H))，7.17(d，J＝3.2Hz，1H)，7.02(d，J＝8.8Hz，1H).

MS(ES+APCI)，m/z：248.0(M+H) ⁺，246.0(M-H) ^-

Embodiment 24

5-chloro-N-(2-chlorphenyl)-2-hydroxybenzamide

(5-chloro-N-(2-chlorophenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，CDCl ₃)，δ11.73(s，1H)，8.45(s，1H)，8.36(d，J＝8.4Hz，1H)，7.52(s，1H)，7.41～7.47(m，2H)，7.34～7.37(m，1H)，7.14～7.18(m，1H)，7.01(d，J＝8.8Hz，1H).

Embodiment 25

5-chloro-N-(3-trifluoromethyl-4-chlorphenyl)-2-hydroxybenzamide

(5-chloro-N-(4-chloro-3-(trifluoromethyl)phenyl)-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.47(s，1H)，10.65(s，1H)，8.29(s，1H)，7.98(d，J＝8.4Hz，1H)，7.85(d，J＝1.6Hz，1H)，7.71(d，J＝8.4Hz，1H)，7.45～7.47(m，1H)，7.01(d，J＝8.8Hz，1H).

Embodiment 26

5-chloro-N-(1H-1,2,4 triazole phenyl)-2-hydroxybenzamide

(N-(4-(1H-1，2，4-triazol-1-yl)phenyl)-5-chloro-2-hydroxybenzamide)

Synthetic method such as embodiment 1.

¹HNMR(400MHz，DMSO-d ₆)，δ11.94(s，1H)，11.74(s，1H)，10.55(s，1H)，9.24(s，1H)，8.21(s，1H)，7.84～7.93(m，4H)，7.46(d，J＝7.6Hz，1H)，7.02(d，J＝8.8Hz，1H).

Embodiment 27

Chemical compound (1 * 10 with variable concentrations ^-8～1 * 10 ^-5M) handle A549 (adenocarcinoma of lung), Du145 (carcinoma of prostate), Hela (cervical cancer), HepG2 (hepatocarcinoma), OS-RC-2 (renal carcinoma), colon cancer tumor cells such as (HT-29) respectively, MTT after 72 hours, hatched again 4 hours, and measured its light absorption value with microplate reader then at 570nm.Found that compound treatment can obviously reduce the absorption of various cells to MTT, illustrate that chemical compound can significantly suppress the propagation of these cells, suppression ratio becomes positive correlation with drug level.According to the growth inhibited effect of chemical compound to these tumor cells, we calculate the half-inhibition concentration (IC of chemical compound to the tumor cell cell ₅₀) be 1 * 10 ^-8～1 * 10 ^-5M.Referring to Fig. 1-Figure 10 and table 1 and table 2.

The IC that table 1. part of compounds suppresses different growth of tumour cell ₅₀(μ M)

The IC that table 2. niclosamide (B037) suppresses different growth of tumour cell ₅₀(μ M)

Cell line	A549 (adenocarcinoma of lung)	L-78 (lung squamous cancer)	Du145 (carcinoma of prostate)	PC3 (carcinoma of prostate)	KB (oral cancer)	A431 (epidermal carcinoma)
							B037 IC50 (μM)	2.8	0.6	0.8	6.7	0.5	8.4
Cell line	MCF-7 (breast carcinoma)	HT-29 (colon cancer)	HT-1080 (human fibrosarcoma)	HepG2 (hepatocarcinoma)	OS-RC-2 (renal carcinoma)	Hela (cervical cancer)
							B037 IC50 (μM)	2	7.4	3	1.1	1.9	1.2

Embodiment 28

Niclosamide (1 * 10 with variable concentrations ^-8～1 * 10 ^-5M) handle Du145 (carcinoma of prostate) cell, getting cell respectively after processing in 12,24,36,48 hours handles with PI (propidium iodide), detect with flow cytometer then, find that niclosamide can significantly prolong the G1 phase of Du145 cell, and the G1 phase is along with the increase of drug level elongated (table 3.).

Table 3. niclosamide (B037) is to the influence of prostate gland cancer cell Du145 cell cycle

Embodiment 29

Tumor cell Du145 or HepG2 cultivate in six orifice plates by the standard cell lines cultural method, and used culture medium is that ATCC recommends to use culture medium, contains 10%FCS.

When cell grows to 80-90%, change culture medium into corresponding serum-free medium, after serum starvation 16-24 hour, add the D147 or the solvent contrast of variable concentrations in the culture medium.After the drug treating two hours, adding EGF (final concentration is 100ng/ml) in culture medium handled after 30 minutes, culture medium is gone in suction, 1XPBS (pH7.2) with 4 ℃ of pre-coolings cleans, the 1XSDS sample buffer cell lysis that in culture hole, adds 0.1ml, sweep cell with cell rapidly, the collecting cell lysate is in the Eppendorf pipe of 1ml, the pair cell lysate carries out supersound process to reduce the viscosity of lysate, subsequently cell pyrolysis liquid is boiled 5 minutes at 95 ℃, makes the thorough degeneration of albumen, put cold after, cell pyrolysis liquid is centrifugal, supernatant is transferred in another Eppenforf pipe, as cell sample liquid.30 μ l cell sample liquid separate through 12.5% SDS-polyacrylamide gel electrophoresis, and the albumen with separator well is transferred on the pvdf membrane through wet method through gel subsequently.After transfer finishes, film with the 1XTBST room temperature sealing that contains 5% defatted milk powder one hour, please be washed 3 times 5 minutes/time with 1XTBST, with anti-(MEK, pMEK, Erk, the pErK of film with suitable dilution, STAT3, pSTAT3, CyclinD3, CyclinE, CyclinB1, Caspase3, Caspase9, GAPDH etc.) diluent applies and to educate, and room temperature or 4 ℃ spend the night, and film are cleaned 3 times with 1XTBST again, 5 minutes/time, subsequently film is hatched room temperature 1 hour with two anti-diluents of the HRP labelling that suitably dilutes.Subsequently film is cleaned 3 times with 1XTBS, 5 minutes/time, press on the handbook of Apharmsham ECL PLUS DETECTION SYSTEM bonded albumen on the ECL detection method detection film then.Western blot test shows that the B037 selectivity reduces the level of Cyclin D3, and to the level of Cyclin E do not exert an influence (Figure 11); The B037 selectivity suppresses the phosphorylation of STAT3, and to the phosphorylation of MAPK signal path node albumen Raf, MEK, ErK do not exert an influence (Figure 12).

1XSDS sample buffer: 62.5mM Tris-HCl (pH6.8), 2%w/v SDS, 10%glycerol, 50mM DTT, 0.01%bromophenol blue

1XTBST：50mM Tris-HCl，pH7.4，150mM NaCl，0.1％Tween-20.

One anti-diluent: 1XTBST contains 0.05%Tween-20,5%BSA.

Two anti-diluent: 1XTBST, 5% defatted milk powder.

Embodiment 30

(cell scratch experiment)

The OS-RC-2 cell inoculation in six orifice plates, is used 1640 culture medium culturings that contain 10%FBS.When the degree of converging of cell reaches the degree of converging of 80%-90%, with the intermediate score of the rifle head of sterilizing at cellular layer.Clean the cell that twice removal suspends with PBS subsequently, reuse contains 0.25%DMSO or contains 0.5 μ M, 1.0 μ M, and 1640 culture medium of 2.5 μ M B037 continue to cultivate 36 hours.Three marked regions by relatively 0 hour and 36 hours each processed group cut districts differ microphotograph, the propagation of analysis B037 pair cell and the influence of transfer ability.The cut repairing test shows that B037 dose dependent ground suppresses the propagation and the migration (Figure 13) of OS-RC-2 tumor cell.

Claims (9)

1. the chemical compound or its pharmaceutically acceptable salt that have formula I, as the application of JAK-STAT3 signal pathway inhibitor in the preparation antitumor drug,

Wherein:

Following a is 0 or 1; B is 0 or 1; N is optional to be 0,1,2 or 3;

Ar is not for containing or contain the aromatic rings of nitrogen, sulfur or oxygen heteroatom;

R ₁Certainly optional:

1)H；

2) halogen (F, Cl, Br, I);

3) O _aC ₁-C ₆Alkyl;

4) O _aC ₁-C ₆Cycloalkyl;

5) O _aC ₁-C ₆Heterocyclic radical;

6)O _aC ₀-C ₈-NR ₃R ₄；

Described alkyl, cycloalkyl and heterocyclic radical are optional to be selected from R by 0 or one or more ₃Substituent group replace;

R ₂Be 0～5 substituent group on the Ar, each substituent group is certainly optional respectively:

1)H；

2) halogen;

3) (CO) _bO _aC ₁-C ₆Alkyl;

4) (CO) _bO _aC ₁-C ₆Cycloalkyl;

5) (CO) _bO _aC ₁-C ₆Heterocyclic radical;

6)(CO) _bO _aC ₀-C ₈-NR ₃R ₄；

7) C ₁-C ₃Perfluor or part fluoro-alkyl

8)(C＝O) _aNR ₆R ₅；

9)OH；

10)CN；

11)NO ₂；

12)SO ₂NR ₃R ₄；

13)COOH；

Described alkyl, cycloalkyl and heterocyclic radical are optional to be replaced by 0 or one or more substituent groups that are selected from R4;

R ₃And R ₄Independently be selected from separately:

1)H；

2) (C=O) _aO _bC ₁-C ₆Alkyl;

3) (C=O) _aO _bAryl;

4) O _bC ₁-C ₃Perfluor or part fluoro-alkyl;

5) (C=O) _aO _bC ₃-C ₆Cycloalkyl;

6) (C=O) _aO _bHeterocyclic radical;

Described alkyl, aryl, cycloalkyl and heterocyclic radical are optional to be selected from R by 0～3 ₃Substituent group replace; Perhaps R ₃And R ₄And the atom that connects them forms the dicyclo that one 4～7 yuan monocycle or each ring are 4～7 yuan, and optionally contains 1～3 hetero atom that is selected from N, O and S, and described monocycle or dicyclo are optional by one or more R that are selected from ₃Substituent group replace.

2, application according to claim 1 is characterized in that, the Ar of described formula I chemical compound is a phenyl.

3, application according to claim 1 is characterized in that, the Ar of described formula I chemical compound is a phenyl, and n is 0.

4, application according to claim 3 is characterized in that, described niclosamide and the pharmaceutically acceptable salt thereof of being applied as is as the application of JAK-STAT3 signal pathway inhibitor in the preparation antitumor drug.

5, application according to claim 1 is characterized in that, described chemical compound has with one of Formula Il～IV structure:

R ₁～R ₄Definition and the R in the claim 1 ₁～R ₄Identical.

According to each described application of claim 1-5, it is characterized in that 6, described have the chemical compound of formula I or the valid density of its pharmaceutically acceptable salt is 1 * 10 ^-8～1 * 10 ^-5M.

7, according to each described application of claim 1-5, it is characterized in that described tumor is any in following: histiocyte lymphatic cancer, nonsmall-cell lung cancer, small cell lung cancer, adenocarcinoma of lung, lung squamous cancer, cancer of pancreas, breast carcinoma, carcinoma of prostate, hepatocarcinoma, skin carcinoma, cell carcinoma, carcinoma of prostate, nasopharyngeal carcinoma, epidermal carcinoma, palace stem cancer, oral cancer, human fibrosarcoma, leukemia.

8, a kind of antitumor Pharmaceutical composition is characterized in that: include the chemical compound shown in the claim 1 Chinese style I or its pharmaceutically acceptable salt and pharmaceutically acceptable carrier.

9, antitumor Pharmaceutical composition according to claim 8 is characterized in that: also include and be selected from the following chemical compound one or more: the medicine and the cell death inducer at estrogenic agents, androgen receptor modifier, retinoid-like receptor modulators, cytotoxin/cytostatics, antiproliferative, protein transferase inhibitor, HMG-CoA reductase inhibitor, HIV kinases inhibitor, reverse transcriptase inhibitors, angiogenesis inhibitor, cell proliferation and the agent of existence signal suppressing, the interference cell cycle outpost of the tax office.

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