CN101343268A - Thiazole couplet pyrazolone series compound and application of the same as Bcl-2 family protein antagonist - Google Patents
Thiazole couplet pyrazolone series compound and application of the same as Bcl-2 family protein antagonist Download PDFInfo
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Abstract
The invention discloses a thiazole bipyrazolone compound as well as applications thereof in preparing a small molecular inhibitor of Bcl-2 family proteins and antitumor drugs, the thiazole bipyrazolone compound has the above structural formula, and the thiazole bipyrazolone compound has the activity for inhibiting the combination of Bcl-xL and natural substrate polypeptide (Bid BH3 polypeptide) in the micromole range, and can serve as the small molecular inhibitor of the Bcl-2 family proteins for inhibiting the anti-apoptosis activity thereof; and the compound shows a certain inhibitory activity to human breast cancer MDA-MB-231 cells and MCF-7 cells, and is expected to be developed into the antitumor drugs of targeted Bcl-2 family proteins.
Description
Technical field
The micromolecular inhibitor that the present invention relates to a kind of thiazole connection pyrazolone compound and work thereof is the application of Bcl-2 family protein antagonist, particularly (E)-4-(2-(aryl hydrazono-)-3-methyl isophthalic acid-(4-aryl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone compounds and application as the Bcl-2 family protein antagonist.This compounds can suppress Bcl-2 family protein member Bcl-x
LWith combining of natural substrate polypeptide (Bid BH3 polypeptide), can be used as the Bcl-2 family protein and be used to suppress its anti-apoptosis activity, and be expected to develop and become the novel antitumor drug of a class.
Background technology
Apoptosis (can be described as programmed cell death under some situation again) is a kind of removal senile cell or paracytic natural death mechanism, the disorder of this mechanism and multiple disease direct relation arranged.Since the nineties in last century, people find tumour gradually be the propagation of cell and apoptosis unbalance due to (Okada, H.; Mak, T.W.Nat.Rev.Cancer 2004,4,592-603).Studies show that apoptosis mechanism is suppressed in kinds of tumor cells, tumour cell thereby be able to hyperplasia; In addition, apoptosis mechanism is suppressed and also makes tumour cell that the resistivity of chemotherapeutics is strengthened (Igney, F.H.; Krammer, P.H.Nat.Rev.Cancer 2002,2,277-288).Therefore, the apoptosis mechanism of managing to recover in the tumour cell becomes the new approaches that current antitumor drug designs, and has obtained extensive attention (Reed, J.C.Nat.Rev.Drug Discov.2002,1,111-121 in the world; Andersen, M.H.; Becker, J.C.; Straten, P.Nat.Rev.Drug Discov.2005,4,399-409.).
With apoptosis-related drug target in, Bcl-2 (B-cell lymphoma 2) associated protein is a class that early obtains studying.This proteinoid can be divided into three families: Bcl-2 family, Bax family and BH3-only family.Wherein, Bcl-2 family member (Bcl-2, Bcl-x
L, Mcl-1, Bcl-w etc.) and play a part anti-apoptotic, the member of latter two family plays short apoptotic effect.It is presently believed that the Bcl-2 associated protein mainly is to play a role in apoptotic plastosome approach.Wherein activated Bax family protein (Bax, Bak etc.) thus can be combined in and make cytochrome C from plastosome, discharge the generation that finally causes apoptosis on the mitochondrial membrane; And Bcl-2, Bcl-x
LThen can combine Deng inhibitor of apoptosis protein, it can not be played a role with Bax and Bak; In addition, some BH3-only family members (Bim, Bad, Bid, Bik etc.) again can with Bcl-2 and Bcl-x
LCombine, suppress its anti-apoptotic effect.Therefore, whether the pair cell apoptosis plays vital regulating and controlling effect to the balance between the Bcl-2 associated protein.Studies show that polytype tumour cell is wherein a kind of Bcl-2 family protein of overexpression at least, but these proteic expression levels are then relatively low in normal cell.Use these proteic functions of organic molecule antagonism to be expected to recover apoptosis mechanism in the tumour cell, thereby reach purpose (Huang, Z.Curr.Opin.Drug Discov.Devel.2000,3, the 565-574 that eliminates tumour; Cory, S.; Adams, J.M.Nat.Rev.Cancer 2002,2,647-656).
Organic molecule detects (Zhang, H. with fluorescence polarization (FluorescencePolarization is called for short FP) method usually to the inhibition of Bcl-2 family protein is active; Nimmer, P.; Rosenberg, S.H.; Ng, S.C.; Joseph, M.Anal.Biochem.2002,307,70-75).This method is based on molecule rotating freely in homogeneous phase solution, when a fluorescently-labeled molecule is excited by a planar polarized light, its emission light can be transmitted into a fixed plane, and the speed of rotation of this radiative polarization level and molecule is inversely proportional to.Fluorescently-labeled small molecules is in the high speed rotating state in homogeneous system, emission light shows as depolarization, obtain a lower polarization value, but not fluorescently-labeled macromole speed of rotation is well below fluorescently-labeled small molecules, after small molecules and the combination of macromole generation specificity, the speed of rotation of mixture is compared variation with macromolecular speed of rotation not obvious, and well below the micromolecular speed of rotation of fluorescent mark, polarization value significantly raises.When this method is used to detect small molecules and suppresses combining of Bcl-2 family protein and natural substrate polypeptide, in the system of fluorescent mark polypeptide and albumen coexistence, add testing compound, if compound also can combine with protein-specific, combine with proteic thereby then can suppress it, and then cause the fluorescence polarization value to reduce with fluorescent mark polypeptide generation competition.Therefore just can record the activity of compound by the changing conditions that detects the fluorescence polarization value.
Summary of the invention
First purpose of the present invention provides a kind of new thiazole connection pyrazolone compound.
Second purpose of the present invention provides a kind of thiazole connection pyrazolone compound and is used for preparation inhibition Bcl-x
LApplication with natural substrate polypeptide bonded micromolecular inhibitor and antitumor drug.
A kind of new thiazole connection pyrazolone compound provided by the present invention has following structural formula:
R in the formula
1Be o-Ph, p-OMe, m-CO
2H, o-CO
2Me or 8-quinoline; R
2Be H, m-Ph, p-Ph, m-Bn, o-OMe, m-OMe, m-NO
2, betanaphthyl, p-N-Pyrrolidine or 8H-indeno [1,2-d] thiazol-2-yl.
Restricted condition is: work as R
1Be m-CO
2During H, R
2Be m-Ph, p-Ph, m-Bn, o-OMe, m-OMe, m-NO
2, betanaphthyl, p-N-Pyrrolidine or 8H-indeno [1,2-d] thiazol-2-yl; Work as R
1Be o-Ph, p-OMe, o-CO
2When Me or 8-quinoline, R
2Be H.
The preparation inhibition Bcl-x that is used for of the present invention
LDo not have above-mentioned restriction with the thiazole of natural substrate polypeptide bonded micromolecular inhibitor and antitumor drug connection pyrazolone compound, specifically have following structural formula (E)-4-(2-(aryl hydrazono-)-3-methyl isophthalic acid-(4-aryl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone compounds:
Formula X
R in the formula
1Be H, m-Me, p-Me, o-Ph, p-F, o-Cl, m-Cl, p-Cl, p-Br, m-OH, p-OMe, p-OEt, m-CH
2OH, o-CO
2H, m-CO
2H, p-CO
2H, o-CO
2Me, p-CONH
2, p-SO
3H, p-SO
2NH
2, p-NMeCOMe, 3,4-dichloro, 2,4,6-tribromo or 8-quinoline; R
2Be H, p-Me, m-Ph, p-Ph, m-Bn, o-OMe, m-OMe, p-OMe, p-Br, m-NO
2, betanaphthyl, p-N-Pyrrolidine or 8H-indeno [1,2-d] thiazol-2-yl.There is not aforesaid restricted condition.Can further describe and be following compound:
Formula X-01, formula X-02, formula X-03,
Formula X-04, formula X-05, formula X-06,
Formula X-07, formula X-08, formula X-09,
Formula X-10, formula X-11, formula X-12,
Formula X-13, formula X-14, formula X-15,
Formula X-16, formula X-17, formula X-18,
Formula X-19, formula X-20, formula X-21,
Formula X-22, formula X-23, formula X-24,
Formula X-25, formula X-26, formula X-27,
Formula X-28, formula X-29, formula X-30,
Formula X-31, formula X-32, formula X-33,
Formula X-34, formula X-35, formula X-36,
Formula X-37, formula X-38, formula X-39,
Formula X-40 or formula X-41
Experiment shows that above-mentioned thiazole connection pyrazolone compound is for suppressing Bcl-2 family protein member Bcl-x
LThe effective constituent that combines with natural substrate polypeptide (Bid BH3 polypeptide) shows that based on the active testing of fluorescence polarization (FP) principle this compounds has the activity of micromole's level.The FP method detects this compounds and suppresses Bcl-x
LWith N end with the Bid BH3 polypeptide of 5-FAM mark (sequence is: 5-FAM-QEDIIRNIARH-LAQVGDSMDR) the bonded activity is as follows:
1) (E)-4-(2-(3-hydroxymethyl phenyl) hydrazono-)-1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl isophthalic acid H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-01) under 10 μ M is 78.7%;
2) (E)-2-(2-(1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-02) under 10 μ M is 77.0%;
3) (E)-3-(2-(1-(4-(4-bromophenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-03) under 10 μ M is 92.4%;
4) (E)-the inhibition constant K of 3-(2-(1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-04)
iBe 0.74 μ M;
5) (E)-3-(2-(1-(4-(biphenyl-3-yl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-05) under 10 μ M is 32.8%;
6) (E)-3-(2-(1-(4-(3-benzyl phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-06) under 10 μ M is 31.4%;
7) (E)-3-(the inhibition constant K of 2-(3-methyl isophthalic acid-(4-(naphthyl-2-) thiazol-2-yl)-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-07)
iBe 2.2 μ M;
8) (E)-4-(2-(3-methyl-5-oxo-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenyl-sulfamide (formula X-08) under 10 μ M is 43.0%;
9) (E)-3-(2-(3-methyl-5-oxo-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-09) under 10 μ M is 35.3%;
10) (E)-the inhibition constant K of 4-(2-(3-methyl-5-oxo-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-10)
iBe 13.1 μ M;
11) (E)-1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-4-(2-phenyl hydrazono-)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-11) under 10 μ M is 68.4%;
12) (E)-4-(2-(1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of Phenylsulfonic acid (formula X-12) under 10 μ M is 56.5%;
13) (E)-4-(2-(4-aminomethyl phenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-13) under 10 μ M is 54.0%;
14) (E)-(4-(2-(1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenyl)-inhibiting rate of N-methylacetamide (formula X-14) under 10 μ M is 48.8% to N-;
15) (E)-(2-(2,4,6-tribromo phenyl) hydrazono-)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-15) under 10 μ M is 43.9% to 3-methyl isophthalic acid-(4-(3-nitrophenyl) thiazol-2-yl)-4-;
16) (E)-4-(2-(1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of benzsulfamide (formula X-16) under 10 μ M is 42.8%;
17) (E)-1-(4-(4-bromophenyl) thiazol-2-yl)-4-(2-(3-chloro-phenyl-) hydrazono-)-3-methyl isophthalic acid H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-17) under 10 μ M is 42.6%;
18) (E)-4-(2-(4-bromophenyl) hydrazono-)-1-(4-(4-bromophenyl) thiazol-2-yl)-3-methyl isophthalic acid H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-18) under 10 μ M is 36.0%;
19) (E)-3-(2-(3-methyl isophthalic acid-(4-(3-nitrophenyl) thiazol-2-yl)-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-19) under 10 μ M is 34.6%;
20) (E)-3-methyl-4-(2-phenyl hydrazono-)-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-20) under 10 μ M is 28.2%;
21) (E)-4-(2-(4-chloro-phenyl-) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-21) under 10 μ M is 26.6%;
22) (E)-4-(2-(2-chloro-phenyl-) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-22) under 10 μ M is 25.5%;
23) (E)-1-(4-(4-bromophenyl) thiazol-2-yl)-4-(2-(3-hydroxyphenyl) hydrazono-)-3-methyl isophthalic acid H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-23) under 10 μ M is 25.4%;
24) (E)-3-(2-(1-(4-(3-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-24) under 10 μ M is 25.3%;
25) (E)-1-(4-(4-bromophenyl) thiazol-2-yl)-3-methyl-4-(2-m-toluene hydrazono-)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-25) under 10 μ M is 23.7%;
26) (E)-4-(2-(4-p-methoxy-phenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-26) under 10 μ M is 22.5%;
27) (E)-3-(2-(3-methyl-5-oxo-1-(4-(4-(Pyrrolidine-1-yl) phenyl) thiazol-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-27) under 10 μ M is 20.5%;
28) (E)-4-(2-(3-methyl-5-oxo-1-(4-p-toluene thiazol-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of benzamide (formula X-28) under 10 μ M is 19.5%;
29) (E)-1-(4-(4-bromophenyl) thiazol-2-yl)-4-(2-(4-p-methoxy-phenyl) hydrazono-)-3-methyl isophthalic acid H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-29) under 10 μ M is 18.9%;
30) (E)-1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-4-(2-m-toluene hydrazono-)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-30) under 10 μ M is 18.0%;
31) (E)-3-(2-(1-(4-(2-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-31) under 10 μ M is 17.5%;
32) (E)-4-(2-(4-fluorophenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-32) under 10 μ M is 14.5%;
33) (E)-4-(2-(3-chloro-phenyl-) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-33) under 10 μ M is 14.2%;
34) (E)-3-(2-(1-(8H-indeno [1,2-d] thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-34) under 10 μ M is 13.5%;
35) (E)-(2-(3, the 4-dichlorophenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-35) under 10 μ M is 13.2% to 4-;
36) (E)-4-(2-(1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of benzamide (formula X-36) under 10 μ M is 8.9%;
37) (E)-3-(2-(1-(4-(biphenyl-4-yl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of phenylformic acid (formula X-37) under 10 μ M is 8.2%;
38) (E)-1-(4-(4-bromophenyl) thiazol-2-yl)-4-(2-(4-ethoxyl phenenyl) hydrazono-)-3-methyl isophthalic acid H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-38) under 10 μ M is 6.3%;
39) (E)-(2-(biphenyl-2-yl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-39) under 10 μ M is 6.3% to 4-;
40) (E)-(2-(quinoline-8-yl) hydrazono-)-1H-pyrazoles-5 (the 4H)-inhibiting rate of ketone (formula X-40) under 10 μ M is 4.4% to 3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-4-;
41) (E)-2-(2-(3-methyl-5-oxo-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) inhibiting rate of methyl benzoate (formula X-41) under 10 μ M is 1.2%.
Thiazole connection pyrazolone compound of the present invention can also be used to prepare antitumor drug.
Experiment shows that thiazole connection pyrazolone compound demonstrates the inhibition activity to human breast carcinoma MDA-MB-231 cell and MCF-7 cell, is that a class has the antitumor lead compound that widespread use is worth.Wherein compounds X-01 and X-15 are to the CC of MDA-MB-231
50Value (50%Cytotoxic Concentration, half cell toxicant concentration) is respectively 3.5 and 0.95 μ M; The CC of compounds X-01 couple MCF-7
50Value is 16.8 μ M.
Beneficial effect of the present invention is that related thiazole connection pyrazolone compound is the Bcl-2 family protein inhibitor of a class formation novelty, molecular level with micromole's level suppresses active, and tumour cell had stronger cytotoxic activity, therefore have good potentiality and application prospect as lead compound, be expected to develop the antitumor drug that becomes target Bcl-2 family protein.
Description of drawings
Fig. 1-Fig. 3 surveys the curve of living for the fluorescence polarization of thiazole connection pyrazolone compound;
Fig. 4-Fig. 6 surveys the curve of living for the MTT of thiazole connection pyrazolone compound.
Wherein, Fig. 4 joins pyrazolone compound X-01 to MDA-MB-231 cell line cell toxicity test for thiazole; Fig. 5 joins pyrazolone compound X-15 to MDA-MB-231 cell line cell toxicity test for thiazole; Fig. 6 joins pyrazolone compound X-01 to MCF-7 cell line cell toxicity test for thiazole.
Embodiment
Embodiment 1: preparation (E)-4-(2-(4-chloro-phenyl-) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone
One, (Z)-2-(2-(4-chloro-phenyl-) hydrazono-)-ethyl 3-oxobutanoate (formula II-05) is synthetic
1.276g (10mmol) 4-chloroaniline (formula I-05) is suspended in the 8mL water, control and slowly inject the dense HCl of 5mL under about room temperature, reduce to and slowly inject the 5mL aqueous solution that contains 0.690g (10mmol) Sodium Nitrite under the low temperature (0 ℃), direct filtration is in the 25mL ethanolic soln that contains 1.28mL (10mmol) methyl aceto acetate and 8g (97.5mmol) sodium acetate afterwards, there is yellow solid to separate out, suction filtration, washing, the dehydrated alcohol recrystallization gets yellow needle-like crystal 1.914g (productive rate 71%).
After testing, structure is correct, and detected result is as follows: mp 80-84 ℃; IR (KBr): v 2994,1704,1617,1587,1528,1495,1474,1456,1419,1396,1372,1330,1276,1205,1167,1116,1093,1021,1008,977,836,818,784,657,512,457cm
-1 1H NMR (300MHz, CDCl
3): δ 14.76 (s, 1H), 7.35 (s, 4H), 4.34 (q, J=7.2Hz, 2H), 2.60 (s, 3H), 1.40 (t, J=7.2Hz, 3H); MS (ESI) is (m/z): 269 (M+H
+), 291 (M+Na
+), 323 (M+MeOH+Na
+), 349 (M+acetone+Na
+); Anal.Calcd for C
12H
13ClN
2O
3: C 53.64, and H 4.88, N 10.43.Found:C 53.60, and H 4.89, N 10.42.
Two, (E)-4-(2-(4-chloro-phenyl-) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-05) synthetic
Dissolve in (Z)-2-(2-(4-chloro-phenyl-) hydrazono-)-ethyl 3-oxobutanoate (formula II-05) adding 15mL acetate with 1.904g (7.08mmol), add 0.646g (7.08mmol) thiosemicarbazide again, reflux 4 hours, cooling back suction filtration, use acetate and refrigerated washing with alcohol successively, directly drain, get yellow solid 1.898g (productive rate 91%).
After testing, structure is correct, and detected result is as follows: mp 215-220 ℃; IR (KBr): v 3375,3264,1693,1600,1549,1481,1446,1397,1313,1288,1269,1234,1087,1045,997,871,823,771,595,547cm
-1 1H NMR (300MHz, DMSO-d
6): δ 13.05 (s, 1H), 9.48 (s, 1H), 8.92 (s, 1H), 7.69 (d, J=9.0Hz, 2H), 7.51 (d, J=8.7Hz, 2H), 2.26 (s, 3H);
13CNMR (75MHz, DMSO-d
6): δ 176.7,156.4, and 149.6,140.4,129.8,129.4,126.7,118.2,11.5; MS (ESI) is (m/z): 296 (M+H
+), 318 (M+Na
+); HRMS (EI) is (m/z): Calcd forC
11H
10ClN
5OS (M
+): 295.0295, Found:295.0294.
Three, (E)-4-(2-(4-chloro-phenyl-) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-21) is synthetic
(E)-4-(2-(4-chloro-phenyl-) hydrazono-)-3-methyl-5-oxo-4 with 0.148g (0.5mmol), 5-dihydro-1 h-pyrazole-1-thioamides (formula III-05) adds in the 15mL dioxane, reflux stops heating to dissolving, adds 0.249g (1.25mmol) 2-bromo-1-(3-benzyl phenyl) ethyl ketone, reflux 0.5h, stop heating, concentrate and remove diox, use sherwood oil and washing with alcohol successively, get compounds X-06 (0.198g, productive rate 100%).
After testing, structure is correct, and detected result is as follows: mp 216-223 ℃; IR (KBr): v 1664,1592,1551,1519,1480,1445,1373,1325,1280,1263,1229,1184,1100,1085,1023,1004,922,884,824,775,728cm
-1 1H NMR (300MHz, CDCl
3): δ 13.25 (s, 1H), 7.96 (dd, J=7.8Hz, J=1.5Hz, 2H), 7.43-7.26 (m, 8H), 2.45 (s, 3H);
13C NMR (75MHz, CDCl
3): δ 156.4,154.9, and 151.8,151.2,139.3,133.9,131.8,129.9,128.5,128.1,127.2,126.4,117.1,107.8,11.9; MS (ESI) is (m/z): 396 (M+H
+), 418 (M+Na
+), 450 (M+MeOH+Na
+); HRMS (ESI) is (m/z): Calcd for C
19H
15ClN
5OS (M+H
+): 396.0686, Found:396.0682.
Embodiment 2: formula X-04, X-05, X-06, X-07, X-09, X-10, X-13, X-19, X-20, X-24, X-26, X-27, X-31, X-32, X-33, X-34, X-35, X-37, X-39, X-40, X-41 compound synthetic
One, formula II-01-04 and formula II-06-17's is synthetic
With embodiment one in like the synthesis type II-05 compounds under the condition, obtain compound formula II-01-04 and formula II-06-17:(Z from corresponding formula I-01-04 and formula I-06-17)-2-(2-phenyl hydrazono-)-ethyl 3-oxobutanoate (formula II-01); (Z)-2-(2-(4-fluorophenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-02); (Z)-2-(2-(2-chloro-phenyl-) hydrazono-)-ethyl 3-oxobutanoate (formula II-03); (Z)-2-(2-(3-chloro-phenyl-) hydrazono-)-ethyl 3-oxobutanoate (formula II-04); (Z)-2-(2-(3, the 4-dichlorophenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-06); (Z)-2-(2-(2-aminomethyl phenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-07); (Z)-2-(2-(3-aminomethyl phenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-08); (Z)-2-(2-(4-aminomethyl phenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-09); (Z)-2-(2-(4-p-methoxy-phenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-10); (Z)-2-(2-(3-carboxyl phenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-11); (Z)-2-(2-(4-carboxyl phenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-12); (Z)-2-(2-(2-methoxycarbonyl phenyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-13); (Z)-2-(2-(the positive butoxy carbonyl phenyl of 4-) hydrazono-)-ethyl 3-oxobutanoate (formula II-14); (Z)-2-(2-(biphenyl-2-yl) hydrazono-)-ethyl 3-oxobutanoate (formula II-15); (Z)-2-(2-(1-naphthyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-16); (Z)-2-(2-(8-quinolyl) hydrazono-)-ethyl 3-oxobutanoate (formula II-17).
Two, formula III-01-04 and formula III-06-17 are synthetic
With embodiment one in like the synthesis type III-05 compounds under the condition, obtain compound formula formula III-01-04 and formula III-06-17:(E)-4-(2-phenyl hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-01) from corresponding formula II-01-04 and formula II-06-17; (E)-and 4-(2-(4-fluorophenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-02); (E)-and 4-(2-(2-chloro-phenyl-) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-03); (E)-and 4-(2-(3-chloro-phenyl-) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-04); (E)-and 4-(2-(3, the 4-dichlorophenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-06); (E)-and 4-(2-(2-aminomethyl phenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-07); (E)-and 4-(2-(3-aminomethyl phenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-08); (E)-and 4-(2-(4-aminomethyl phenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-09); (E)-and 4-(2-(4-p-methoxy-phenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-10); (E)-and 4-(2-(3-carboxyl phenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-11); (E)-and 4-(2-(4-carboxyl phenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-12); (E)-and 4-(2-(2-methoxycarbonyl phenyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-13); (E)-and 4-(2-(the positive butoxy carbonyl phenyl of 4-) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-14); (E)-and 4-(2-(biphenyl-2-yl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-15); (E)-and 4-(2-(1-naphthyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-16); (E)-and 4-(2-(8-quinolyl) hydrazono-)-3-methyl-5-oxo-4,5-dihydro-1 h-pyrazole-1-thioamides (formula III-17).
Three, formula X-04, X-05, X-06, X-07, X-09, X-10, X-13, X-19, X-20, X-24, X-26, X-27, X-31, X-32, X-33, X-34, X-35, X-37, X-39, X-40, X-41 compound synthetic
With embodiment one in like the synthesis type X-21 compounds under the condition, obtain compound formula X-04 from corresponding formula III-01-04 and formula III-06-17 and formula IV-01-04 and formula IV-06-17, X-05, X-06, X-07, X-09, X-10, X-13, X-19, X-20, X-24, X-26, X-27, X-31, X-32, X-33, X-34, X-35, X-37, X-39, X-40, X-41:(E)-3-(2-(1-(4-(4-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-04), (E)-3-(2-(1-(4-(biphenyl-3-yl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-05), (E)-3-(2-(1-(4-(3-benzyl phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-06), (E)-3-(2-(3-methyl isophthalic acid-(4-(naphthyl-2-) thiazol-2-yl)-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-07), (E)-3-(2-(3-methyl-5-oxo-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-09), (E)-4-(2-(3-methyl-5-oxo-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-10), (E)-4-(2-(4-aminomethyl phenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-13), (E)-3-(2-(3-methyl isophthalic acid-(4-(3-nitrophenyl) thiazol-2-yl)-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-19), (E)-3-methyl-4-(2-phenyl hydrazono-)-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-20), (E)-3-(2-(1-(4-(3-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-24), (E)-4-(2-(4-p-methoxy-phenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-26), (E)-3-(2-(3-methyl-5-oxo-1-(4-(4-(Pyrrolidine-1-yl) phenyl) thiazol-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-27), (E)-3-(2-(1-(4-(2-p-methoxy-phenyl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-31), (E)-4-(2-(4-fluorophenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-32), (E)-4-(2-(3-chloro-phenyl-) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-33), (E)-3-(2-(1-(8H-indeno [1,2-d] thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-34), (E)-(2-(3 for 4-, the 4-dichlorophenyl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-35), (E)-3-(2-(1-(4-(biphenyl-4-yl) thiazol-2-yl)-3-methyl-5-oxo-1H-pyrazoles-4 (5H)-subunit) diazanyl) phenylformic acid (formula X-37), (E)-4-(2-(biphenyl-2-yl) hydrazono-)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-1H-pyrazoles-5 (4H)-ketone (formula X-39), (E)-3-methyl isophthalic acid-(4-phenyl thiazole-2-yl)-4-(2-(quinoline-8-yl) hydrazono-)-1H-pyrazoles-5 (4H)-ketone (formula X-40), (E)-2-(2-(3-methyl-5-oxo-1-(4-phenyl thiazole-2-yl)-1H-pyrazoles-4 (5H)-subunit) diazanyl) methyl benzoate (formula X-41).
Embodiment 3: thiazole connection pyrazolone compound is to Bcl-x
LInhibiting fluorescence polarization detects
The Bcl-x that glutathione-S-transferase (GST) merges
L(GST-Bcl-x
L) albumen expresses in e. coli bl21, through the method separation and purification of glutathione agarose gel column affinity chromatography; The fluorescent mark polypeptide is a N end, and (sequence is: 5-FAM-QEDIIRNIARHLAQVGDSMDR) with the Bid BH3 polypeptide of 5-FAM mark; Compounds X-04, X-05, X-06, X-07, X-09, X-10, X-13, X-19, X-20, X-21, X-24, X-26, X-27, X-31, X-32, X-33, X-34, X-35, X-37, X-39, X-40, the X-41 method for preparing, all the other compounds are available from SPECS company.
One, compound formula X-04, the inhibition constant measuring of X-07 and X-10
(1 * phosphate buffer soln contains 0.02% (w/v) NaN in assay buffer
3) the middle GST-Bcl-x that adds
L1 * PBS solution and the DMSO solution of compound different concns, the room temperature lucifuge is hatched 30min behind the mixing; Add 1 * PBS solution (final concentration is 10nM) of fluorescent mark polypeptide again, make the cumulative volume of each solution be 200 μ L, the room temperature lucifuge is hatched 20min behind the mixing; Above-mentioned solution and calibration solution 1 (1nM fluorescein) are respectively got 60 μ L with calibration solution 2 (10mM NaOH) be transferred to (parallel three groups) in black 384 orifice plates, on microplate reader, carry out the detection of fluorescence polarization immediately, with 485nM is excitation wavelength, 535nM is an emission wavelength, the fluorescence polarization value of calibration solution is decided to be 20mP, record the fluorescence polarization value (mP) under each concentration and calculate inhibiting rate, match obtains half-inhibition concentration IC
50, suppress constant and use following formula to try to achieve: K
i=[I]
50/ ([L]
50/ K
d+ [P]
0/ K
d+ 1) ([I]
50: the concentration of free inhibitor when reaching the half inhibition; [L]
50: the concentration of free fluorescent mark polypeptide when reaching the half inhibition; [P]
0: free protein concentration when not suppressing; K
d: the dissociation constant of albumen and fluorescent mark polypeptide).The results are shown in Table 1-table 3:
Table 1: the fluorescence polarization detected result of compounds X-04
Table 2: the fluorescence polarization detected result of compounds X-07
Table 3: the fluorescence polarization detected result of compounds X-10
Two, compound formula X-01-X-03, X-05, X-06, X-08, X-09, the inhibition of X-11-X-41 is active to be detected
(1 * phosphate buffer soln contains 0.02% (w/v) NaN in assay buffer
3) the middle GST-Bcl-x that adds
L1 * PBS solution, add the DMSO solution (final concentration is 10 μ M) of DMSO (2 μ L) and testing compound respectively, the room temperature lucifuge is hatched 30min behind the mixing; Add 1 * PBS solution (final concentration is 10nM) of fluorescent mark polypeptide again, make the cumulative volume of each solution be 200 μ L, the room temperature lucifuge is hatched 20min behind the mixing; Above-mentioned solution and calibration solution 1 (1nM fluorescein) are respectively got 60 μ L with calibration solution 2 (10mM NaOH) be transferred to (parallel three groups) in black 384 orifice plates, on microplate reader, carry out the detection of fluorescence polarization immediately, with 485nM is excitation wavelength, 535nM is an emission wavelength, the fluorescence polarization value of calibration solution is decided to be 20mP, records the fluorescence polarization value (mP) of each compound.
The results are shown in Table 4:
Table 4: compound formula X-01-X-03, X-05, X-06, X-08, X-09, the fluorescence polarization detected result of X-11-X-41
| Compound | GST-Bcl-x LConcentration | GST-Bcl-x LDissociation constant K with the fluorescent mark polypeptide d | Contrast mP ± SD | Fluorescent mark polypeptide mP ± SD | Compound mP ± SD | Inhibiting rate % |
| X-0 1 | 160nM | 78.7nM | 175.93±5.93 | 117.87±7.90 | 130.23±0.63 | 78. 7 |
| X-0 2 | 120nM | 51.3nM | 237.88±16.7 7 | 151.92±53.5 9 | 171.65±22.0 6 | 77. 0 |
| |
160nM | 78.7nM | 175.93±5.93 | 117.87±7.90 | 122.29±2.15 | 92. 4 |
| |
132nM | 55.3nM | 148.37±4.24 | 82.32±6.13 | 126.69±0.11 | 32. 8 |
| |
132nM | 55.3nM | 148.37±4.24 | 82.32±6.13 | 127.63±1.67 | 31. 4 |
| |
87.5nM | 75nM | 152.84±3.59 | 109.38±15.7 6 | 134.15±4.26 | 43. 0 |
| |
132nM | 55.3nM | 183.18±1.96 | 108.77±8.12 | 156.93±3.32 | 35. 3 |
| X-1 1 | 160nM | 78.7nM | 175.93±5.93 | 117.87±7.90 | 136.22±0.25 | 68. 4 |
| X-1 2 | 160nM | 78.7nM | 168.93±12.8 5 | 99.44±14.63 | 129.64±2.49 | 56. 5 |
| |
160nM | 78.7nM | 158.48±4.47 | 128.71±15.5 1 | 142.41±5.72 | 54. 0 |
| |
160nM | 78.7nM | 158.48±4.47 | 128.71±15.5 1 | 143.96±2.85 | 48. 8 |
| |
87.5nM | 75nM | 152.84±3.59 | 109.38±15.7 6 | 133.74±2.20 | 43. 9 |
| |
160nM | 78.7nM | 168.93±12.8 5 | 99.44±14.63 | 139.18±4.01 | 42. 8 |
| |
160nM | 78.7nM | 158.48±4.47 | 128.71±15.5 1 | 145.79±4.34 | 42. 6 |
| |
160nM | 78.7nM | 153.85±7.37 | 105.76±14.6 5 | 136.53±3.66 | 36. 0 |
| |
132nM | 55.3nM | 168.49±3.18 | 65.25±7.07 | 132.78±1.84 | 34. 6 |
| X-2 0 | 132nM | 55.3nM | 183.18±1.96 | 108.77±8.11 | 162.21±3.12 | 28. 2 |
| X-2 1 | 160nM | 78.7nM | 158.48±4.47 | 128.71±15.5 1 | 150.57±4.92 | 26. 6 |
| X-2 2 | 160nM | 78.7nM | 175.93±5.93 | 117.87±7.90 | 161.1±11.04 | 25. 5 |
| |
160nM | 78.7nM | 153.85±7.37 | 105.76±14.6 5 | 141.62±2.93 | 25. 4 |
| |
132nM | 55.3nM | 148.37±4.24 | 82.319±6.13 | 131.69±4.03 | 25. 3 |
| |
160nM | 78.7nM | 168.93±12.8 5 | 99.44±14.63 | 152.48±10.7 5 | 23. 7 |
| |
132nM | 55.3nM | 183.18±1.96 | 108.77±8.11 | 166.46±1.44 | 22. 5 |
| |
132nM | 55.3nM | 148.37±4.24 | 82.32±6.13 | 134.8±0.24 | 20. 5 |
| |
87.5nM | 75nM | 179.4±1.91 | 92.84±15.08 | 162.51±2.73 | 19. 5 |
| |
160nM | 78.7nM | 158.48±4.47 | 128.71±15.5 1 | 152.86±3.92 | 18. 9 |
| X-3 0 | 160nM | 78.7nM | 168.93±12.8 5 | 99.44±14.63 | 156.42±12.1 5 | 18. 0 |
| X-3 1 | 132nM | 55.3nM | 168.49±3.18 | 65.25±7.07 | 150.38±0.23 | 17. 5 |
| X-3 2 | 132nnM | 55.3nM | 183.18±1.96 | 108.77±8.11 | 172.39±4.41 | 14. 5 |
| |
132nM | 55.3nM | 163.99±0.92 | 75.73±7.93 | 151.45±3.22 | 14. 2 |
| |
132nM | 55.3nM | 148.37±4.24 | 82.32±6.13 | 139.44±0.51 | 13. 5 |
| |
132nM | 55.3nM | 163.99±0.92 | 75.73±7.93 | 152.33±3.76 | 13. 2 |
| |
160nM | 78.7nM | 168.93±12.8 5 | 99.44±14.63 | 162.76±0.22 | 8.9 |
| |
132nM | 55.3nM | 148.37±4.24 | 82.32±6.13 | 142.98±4.33 | 8.2 |
| |
160nM | 78.7nM | 158.48±4.47 | 128.71±15.5 1 | 156.59±2.84 | 6.3 |
| |
132nM | 55.3nM | 183.18±1.96 | 108.77±8.11 | 178.49±2.15 | 6.3 |
| X-4 0 | 132nM | 55.3nM | 163.99±0.92 | 75.73±7.93 | 160.06±0.40 | 4.4 |
| X-4 1 | 132nM | 55.3nM | 163.99±0.92 | 75.73±7.93 | 162.95±2.57 | 1.2 |
Embodiment 4: thiazole connection pyrazolone compound detects the cytotoxicity of human breast carcinoma MDA-MB-231 clone
One, compounds X-01, X-02, X-15 measures the MDA-MB-231 cell inhibitory rate under 10 μ M
4 * 10
4The every hole of/ml MDA-MB-231 cell suspension 100 μ L, hatch one day after, mix (final concentration 10 μ M), 37 ℃, 5%CO with different testing compound solution
2Cultivated 2 days, and adopted the MTT colorimetry to detect cytotoxicity.The multi-functional microplate reader of 570nm TECAN GENios Pro is measured the OD value, calculates cell inhibitory rate.
Table 5: compounds X-01, X-02, X-15 measures the MDA-MB-231 cell inhibitory rate under 10 μ M
Two, the cytotoxicity of compounds X-01 couple MDA-MB-231 detects
4 * 10
4The every hole of/ml MDA-MB-231 cell suspension 100 μ L, hatch one day after, add the DMEM nutrient solution mixed the different concns compound, 37 ℃, 5%CO
2Cultivated 2 days, and adopted the MTT colorimetry to detect cytotoxicity.The multi-functional microplate reader of 570nm TECAN GENios Pro is measured the OD value, calculates CC
50Value (50%Cytotoxic Concentration) is 3.5 μ M.
Table 6: compounds X-01 pair MDA-MB-231 cytotoxicity
Three, the cytotoxicity of compounds X-15 couple MDA-MB-231 detects
4 * 10
4The every hole of/ml MDA-MB-231 cell suspension 100 μ L, hatch one day after, add the DMEM nutrient solution mixed the different concns compound, 37 ℃, 5%CO
2Cultivated 2 days, and adopted the MTT colorimetry to detect cytotoxicity.The multi-functional microplate reader of 570nm TECAN GENios Pro is measured the OD value, calculates CC
50Value (50%Cytotoxic Concentration) is 0.95 μ M.
Table 7: compounds X-15 pair MDA-MB-231 cytotoxicity
Embodiment 5: thiazole connection pyrazolone compound detects the cytotoxicity of human breast carcinoma MCF-7 clone
One, compounds X-01, X-02, X-03, X-11, X-15 measures the MCF-7 cell inhibitory rate under 10 μ M
6 * 10
4The every hole of/ml MCF-7 cell suspension 100 μ L, hatch one day after, mix (final concentration 10 μ M), 37 ℃, 5%CO with different testing compound solution
2Cultivated 4 days, and adopted the MTT colorimetry to detect cytotoxicity.The multi-functional microplate reader of 570nm TECAN GENios Pro is measured the OD value, calculates cell inhibitory rate.
Table 8: compounds X-01, X-02, X-03, X-11, X-15 measures the MCF-7 cell inhibitory rate under 10 μ M
Two, the cytotoxicity of compounds X-01 couple MCF-7 detects
6 * 10
4The every hole of/ml MCF-7 cell suspension 100 μ L, hatch one day after, add the DMEM nutrient solution mixed the different concns compound, 37 ℃, 5%CO
2Cultivated 4 days, and adopted the MTT colorimetry to detect cytotoxicity.The multi-functional microplate reader of 570nm TECAN GENios Pro is measured the OD value, calculates CC
50Value (50%Cytotoxic Concentration) is 16.8 μ M.
Table 9: compounds X-01 pair MCF-7 cytotoxicity
Claims (3)
1, class thiazole connection pyrazolone compound is characterized in that having following structural formula:
R in the formula
1Be o-Ph, p-OMe, m-CO
2H, o-CO
2Me or 8-quinoline; R
2Be H, m-Ph, p-Ph, m-Bn, o-OMe, m-OMe, m-NO
2, betanaphthyl, p-N-Pyrrolidine or 8H-indeno [1,2-d] thiazol-2-yl.
Restricted condition is: work as R
1Be m-CO
2During H, R
2Be m-Ph, p-Ph, m-Bn, o-OMe, m-OMe, m-NO
2, betanaphthyl, p-N-Pyrrolidine or 8H-indeno [1,2-d] thiazol-2-yl; Work as R
1Be o-Ph, p-OMe, o-CO
2When Me or 8-quinoline, R
2Be H.
2, a kind of thiazole connection pyrazolone compound is used to prepare the application of the micromolecular inhibitor and the antitumor drug of Bcl-2 family protein, and described thiazole connection pyrazolone compound has following structural formula:
R in the formula
1Be H, m-Me, p-Me, o-Ph, p-F, o-Cl, m-Cl, p-Cl, p-Br, m-OH, p-OMe, p-OEt, m-CH
2OH, o-CO
2H, m-CO
2H, p-CO
2H, o-CO
2Me, p-CONH
2, p-SO
3H, p-SO
2NH
2, p-NMeCOMe, 3,4-dichloro, 2,4,6-tribromo or 8-quinoline; R
2Be H, p-Me, m-Ph, p-Ph, m-Bn, o-OMe, m-OMe, p-OMe, p-Br, m-NO
2, betanaphthyl, p-N-Pyrrolidine or 8H-indeno [1,2-d] thiazol-2-yl.
3, the application of class thiazole connection pyrazolone compound as claimed in claim 2 is characterized in that described Bcl-2 family protein is Bcl-x
L, Bcl-2 or Mcl-1.
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| CN103304559A (en) * | 2012-03-06 | 2013-09-18 | 南京大学 | Thiazole derivatives comprising pyrazoline and thiophene (or furan) structures, and preparation method and application thereof |
| EP2766355A4 (en) * | 2011-10-11 | 2015-06-17 | Dana Farber Cancer Inst Inc | Pyrazol-3-ones that activate pro-apoptotic bax |
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| CN101766602B (en) * | 2008-12-30 | 2012-01-11 | 中国医学科学院血液病医院（血液学研究所） | Application of substituted aryl hydrazone compound serving as anti-tumor necrosis factor inhibitor medicament |
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| EP2766355A4 (en) * | 2011-10-11 | 2015-06-17 | Dana Farber Cancer Inst Inc | Pyrazol-3-ones that activate pro-apoptotic bax |
| US9303024B2 (en) | 2011-10-11 | 2016-04-05 | Dana-Farber Cancer Institute, Inc. | Pyrazol-3-ones that activate pro-apoptotic BAX |
| US10000478B2 (en) | 2011-10-11 | 2018-06-19 | Dana-Farber Cancer Institute, Inc. | Pyrazol-3-ones that activate pro-apoptotic BAX |
| AU2017228527B2 (en) * | 2011-10-11 | 2019-02-21 | Dana-Farber Cancer Institute, Inc. | Pyrazol-3-ones that activate pro-apoptotic bax |
| US10351554B2 (en) | 2011-10-11 | 2019-07-16 | Dana-Farber Cancer Institute, Inc. | Pyrazol-3-ones that activate pro-apoptotic BAX |
| US10844053B2 (en) | 2011-10-11 | 2020-11-24 | Dana-Farber Cancer Institute, Inc. | Pyrazol-3-ones that activate pro-apoptotic BAX |
| EP3799871A3 (en) * | 2011-10-11 | 2021-06-16 | Dana Farber Cancer Institute, Inc. | Pyrazol-3-ones that activate pro-apototic bax |
| US11358960B2 (en) | 2011-10-11 | 2022-06-14 | Dana-Farber Cancer Institute, Inc. | Pyrazol-3-ones that activate pro-apoptotic BAX |
| CN103304559A (en) * | 2012-03-06 | 2013-09-18 | 南京大学 | Thiazole derivatives comprising pyrazoline and thiophene (or furan) structures, and preparation method and application thereof |
| CN103304559B (en) * | 2012-03-06 | 2016-05-18 | 南京大学 | One class is containing thiazole derivative and method for making and the purposes of pyrazoline and thiophene (or furans) structure |
| EP3630099A4 (en) * | 2017-06-01 | 2021-07-14 | Albert Einstein College of Medicine | Bax activators and uses thereof in cancer therapy |
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