CN110407767B - Stilbene analogue containing thiazole ring structure and synthesis method and application thereof - Google Patents
Stilbene analogue containing thiazole ring structure and synthesis method and application thereof Download PDFInfo
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- C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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Abstract
The invention disclosesA Stilbene analogue containing a thiazole ring structure and a synthesis method and application thereof are disclosed, wherein the structural formula of the Stilbene analogue containing the thiazole ring structure is shown as a formula (I):
Description
Technical Field
The invention relates to a Stilbene analogue containing a thiazole ring structure, and a synthesis method and application thereof.
Background
Stilbene compounds are a generic name of compounds with Stilbene as structural parent nucleus, and widely exist in nature as phytoalexins, such as Gnetum microphyllum of Gnetaceae, Veratrum nigrum of Liliaceae, Polygonum cuspidatum and Polygonum multiflorum of Polygonaceae, mulberry of Moraceae, grape of Vitaceae, etc. Stilbene compounds have wide biological activity and are widely concerned, and in the field of medicine, the stilbene compounds have activities of antibiosis and anti-inflammation (J, obtained from the company of Food, 2017, 65(51): 11179-; in the pesticide field, the pesticide composition has the activities of resisting fungi (Plant Dis., 2019, 103(7): 1674-. As can be seen from the above, stilbene compounds have wide biological activity, and are ideal potential drug lead structures due to simple structure and natural source skeleton structure.
The nitrogenous heterocyclic compound has the characteristics of high target specificity and good environmental compatibility due to the similarity of an alkaloid structure in an organism, and becomes the mainstream research field for creating new medicaments. Wherein, the thiazole compound has wide biological activity due to the unique aromatic heterocyclic structure, such as medical activity of cancer resistance (Eur. J. Med. chem., 2015, 92(6): 866-875), inflammation resistance (Comut. biol. chem., 2016, 61: 86-96), bacteria resistance (bioorg. Med. chem. Lett., 2012, 22(24): 7719-7725, J. Antibiot., 2014, 68(4): 259-66), oxidation resistance (Chemistry Select, 2019, 4(19): 5570-5576) and the like; and herbicidal activities such as herbicidal (organic chemistry, 2009, 29(6): 924-928), fungicidal (J. enzym. inhib. Med. CH., 2019, 34(1): 898-908), insecticidal (J. agric. Food Chem, 2011, 59(9): 2932-2937). According to literature reports (WO 2001068589, 2001-09-20; WO 2004035554, 2004-04-29) and previous work of the applicant (organic chemistry, 2009, 29(12): 2000-2004), it is shown that fluorine-containing phenylthiazoles are also a class of structural units with good biological activity.
In view of the fact that stilbene compounds and thiazole heterocyclic compounds have good biological activity, the invention utilizes the method of active substructure splicing and biological isostere substitution to substitute a benzene ring in a stilbene skeleton with fluorine-containing phenylthiazole, designs and synthesizes a novel stilbene analogue containing a thiazole ring structure, and aims to find out the new biological activity of the novel stilbene analogue.
The structure and bioactivity research of the series of stilbene analogs containing thiazole ring structures designed and synthesized by the invention is not reported in documents.
Disclosure of Invention
In order to solve the technical problems in the prior art, the invention aims to provide a Stilbene analogue containing a thiazole ring structure, a synthesis method and an application thereof.
The Stilbene analogue containing a thiazole ring structure is characterized in that the structural formula is shown as the formula (I):
in the formula (I), H on a benzene ring is mono-substituted, di-substituted or not substituted by a substituent R; n is an integer of 0-2, and n represents the number of substituent groups R on a benzene ring; n =0, represents that H on the phenyl ring is unsubstituted; n =1, represents H on the phenyl ring monosubstituted by a substituent R; n =2, it represents that H on the benzene ring is disubstituted by a substituent R, and the substituents R on different substitution positions are the same or different; the substituent R is hydrogen, alkyl of C1-C4, halogenated alkyl of C1-C3, alkoxy of C1-C3 or halogen.
The Stilbene analogue containing a thiazole ring structure is characterized in that in the formula (I), a substituent R is hydrogen, methyl, methoxy, tert-butyl, trifluoromethyl, F, Cl or Br.
The Stilbene analogue containing a thiazole ring structure is characterized in that in the formula (I), R (n) is hydrogen, o-methyl, m-methyl, p-methyl, o-methoxy, p-tert-butyl, p-methoxy, m-methoxy, p-trifluoromethyl, p-fluorine, o-chlorine, p-chlorine, o-bromine, m-bromine or 2, 4-dichloro.
The synthesis method of the Stilbene analogue containing the thiazole ring structure is characterized by comprising the following steps:
1) mixing 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole shown as a formula (II) and triethyl phosphite shown as a formula (III) in a molar ratio of 1: 1.5-5.0, then reacting under a heating condition, monitoring by TLC (thin layer chromatography) until the reaction is finished, and concentrating to remove redundant triethyl phosphite to obtain a concentrated solution;
2) adding a solvent DMF, sodium hydroxide and substituted benzaldehyde shown as a formula (IV) into the concentrated solution obtained in the step 1), reacting at room temperature, monitoring by TLC (thin layer chromatography) until the reaction is finished, and performing post-treatment on the reaction solution to obtain a Stilbene analogue containing a thiazole ring structure shown as a formula (I);
in the formula (IV), H on a benzene ring is mono-substituted, di-substituted or not substituted by a substituent R; n is an integer of 0-2, and n represents the number of substituent groups R on a benzene ring; n =0, indicates that H on the phenyl ring is not substituted; n =1, represents H on the phenyl ring monosubstituted by a substituent R; n =2, it represents that H on the benzene ring is disubstituted by substituent R, and the substituents R on different substitution positions are the same or different; the substituent R is hydrogen, alkyl of C1-C4, halogenated alkyl of C1-C3, alkoxy of C1-C3 or halogen.
The method for synthesizing the Stilbene analogue containing the thiazole ring structure is characterized in that the ratio of the amounts of the 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole shown as the formula (II), the substituted benzaldehyde shown as the formula (IV) and the sodium hydroxide is 1: 1.0-3.0: 1.0-5.0.
The synthesis method of the Stilbene analogue containing a thiazole ring structure is characterized in that the molar ratio of 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole shown as a formula (II) to a solvent DMF is 1: 2.0-20, and preferably 1: 4.0-10.
The method for synthesizing the Stilbene analogue containing the thiazole ring structure is characterized in that in the step 1), the heating reaction temperature is 100-130 ℃, the heating reaction time is 1-3 hours, and in the step 2), the reaction time at room temperature is 3-5 hours.
The method for synthesizing the Stilbene analogue containing the thiazole ring structure is characterized in that in the step 2), the step of post-treating the reaction liquid is as follows: after the reaction is finished, adding a large amount of ice water into the reaction liquid, stirring, if solid is precipitated, filtering, and recrystallizing and purifying a filter cake by using an organic solvent to obtain the Stilbene analogue containing the thiazole ring structure shown in the formula (I); if no solid is precipitated, extracting with ethyl acetate, desolventizing, and separating the desolventized residual liquid by column chromatography to obtain the Stilbene analogue containing the thiazole ring structure shown in the formula (I); wherein the mass ratio of the added mass of the ice water to the 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole added in the step 1) is 33-50: 1.
The method for synthesizing the Stilbene analogue containing the thiazole ring structure is characterized in that the organic solvent used for recrystallization is one or a mixture of more than two of ethanol, ethyl acetate and n-hexane; the eluent used for column chromatography separation is a mixed solution of ethyl acetate and petroleum ether in a volume ratio of 1: 3-8.
The Stilbene analogue containing a thiazole ring structure is applied to preparation of antitumor drugs.
In the process of preparing the thilbene analogue containing a thiazole ring structure, the amount of triethyl phosphite and the reaction temperature must be controlled within a certain range, because triethyl phosphite has certain reducibility, and if the amount of triethyl phosphite is large and the reaction temperature is high (reflux temperature), bromine at the 5-position of the thiazole ring in the thilbene analogue containing the thiazole ring structure can be removed, so that the structural formula of the target product is changed.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a novel Stilbene analogue containing a thiazole ring structure, which has a simple preparation process and shows certain antitumor activity. In the embodiment of the invention, the synthetic compound is subjected to TOP I (DNA topoisomerase) inhibition activity determination by using a DNA loosening experiment, and the result of the DNA loosening experiment shows that the synthetic compound has certain TOP I inhibition activity at a concentration of 50 mu M, wherein the inhibition activity of compound Ib on the compound is about 70%.
Detailed Description
The invention is further illustrated with reference to the following specific examples, without limiting the scope of the invention thereto.
Example 1 synthesis of compound Ia (r (n) = H):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (2.5g, 15mmol), heated to 100 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 3 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (2.9g, 40mmol), benzaldehyde (1.1g, 10mmol) and sodium hydroxide (0.4 g, 10mmol) and reacted at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), finishing the reaction for about 4h, then pouring the reaction liquid into ice water (120 mL), stirring, separating out a solid, filtering, and recrystallizing a filter cake by using ethyl acetate to obtain 1.9 g of a yellow solid, namely the (E) -5-bromo-4- (4-fluorophenyl) -2-styrylthiazole, wherein the calculated yield is 53.2%. m.p. 108-113 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.96 (dd, J = 8.5, 5.5 Hz, 2H), 7.55 (d, J = 8.5 Hz, 2H), 7.46 – 7.33 (m, 4H), 7.26 (d, J = 16.0 Hz, 1H), 7.17 (t, J = 8.5 Hz, 2H)；
HRMS (ESI) calcd C 17 H 11 BrFNS [M+H] + 359.9852, found 359.9836。
example 2 synthesis of compound Ib (r (n) = o-chloro):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (3.3g, 20mmol), heated to 120 ℃ for reaction, and the progress of the reaction was checked by TLC, after about 1.5 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (4.0g, 55mmol), o-chlorobenzaldehyde (1.4g, 10mmol) and sodium hydroxide (1.3 g, 32 mmol) to react at room temperature. The reaction process is detected by TLC (thin layer chromatography), about 4.5h is finished, then the reaction liquid is poured into ice water (130 mL), stirred, solid is separated out, filtered, and a filter cake is recrystallized by using normal hexane to obtain 1.86 g of yellow solid, namely the (E) -2- (2-chlorostyryl) -5-bromo-4- (4-fluorophenyl) thiazole, and the calculated yield is 47.3%. m.p. 124-127 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.96 (dd, J = 9.0, 5.5 Hz, 2H), 7.74 (d, J = 16.0 Hz, 1H), 7.68 (dd, J = 7.5, 2.5 Hz, 1H), 7.43 (dd, J = 7.5, 2.5 Hz, 1H), 7.34 – 7.23 (m，3H), 7.17 (t, J = 8.5 Hz, 2H)；
HRMS (ESI) calcd C 17 H 10 BrClFNS [M+H] + 393.9463, found 393.9451。
example 3 synthesis of compound Ic (r (n) = p-chloro):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (2.8g, 17mmol), heated to 130 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 1 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (3.7g, 50mmol), p-chlorobenzaldehyde (1.7g, 12 mmol) and sodium hydroxide (0.9 g, 22 mmol) to react at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), finishing the reaction for about 4h, then pouring the reaction liquid into ice water (185 mL), stirring, separating out a solid, filtering, and recrystallizing a filter cake by using n-hexane to obtain 2.4g of a yellow solid, namely (E) -2- (4-chlorostyryl) -5-bromo-4- (4-fluorophenyl) thiazole, wherein the yield is calculated to be 61.0%. m.p. 148-151 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.94 (dd, J = 9.0, 5.5 Hz, 2H), 7.47 (d, J = 8.5 Hz, 2H), 7.37 (d, J = 8.5 Hz, 2H), 7.31 (d, J = 16.5 Hz, 1H), 7.20 (d, J = 16.5 Hz, 1H), 7.16 (t, J = 9.0 Hz, 2H);
HRMS (ESI) calcd C 17 H 10 BrClFNS [M+H] + 393.9463, found 393.9439。
example 4 synthesis of compound Id (r (n) = p-fluoro):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (8.25 g, 50mmol), heated to 120 ℃ for reaction, and the progress of the reaction was monitored by TLC, after about 2 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (4.4g, 60 mmol), p-fluorobenzaldehyde (1.2g, 10mmol) and sodium hydroxide (0.4 g, 10mmol) and reacted at room temperature. The reaction process is detected by TLC, about 3h after the reaction is finished, the reaction liquid is poured into ice water (170 mL), stirred, solid is separated out, filtered, and filter cake is recrystallized by ethyl acetate to obtain 1.8g of yellow solid, namely (E) -2- (4-fluorostyryl) -5-bromo-4- (4-fluorophenyl) thiazole, and the yield is calculated to be 48.7%. m.p. is 132-135 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.95 (dd, J = 8.5, 5.5 Hz, 2H), 7.52 (dd, J = 8.5, 5.5 Hz, 2H), 7.32 (d, J = 16.0 Hz, 1H), 7.20 – 7.12 (m, 3H), 7.10 (t, J = 8.5 Hz, 2H)；
HRMS (ESI) calcd C 17 H 10 BrF 2 NS [M+H] + 377.9758, found 377.9802。
example 5 synthesis of compound Ie (r (n) = p-methoxy):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (3.3g, 20mmol), heated to 115 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 1.5 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (4.4g, 60 mmol), p-methoxybenzaldehyde (2.7g, 20mmol) and sodium hydroxide (0.8 g, 20mmol) and reacted at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), finishing the reaction for about 3h, then pouring the reaction liquid into ice water (150 mL), stirring, separating out a solid, filtering, and recrystallizing a filter cake by using ethanol to obtain 1.8g of a yellow solid, namely (E) -2- (4-methoxystyryl) -5-bromo-4- (4-fluorophenyl) thiazole, wherein the calculated yield is 61.3%. m.p. 145-146 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.95 (dd, J = 8.5, 5.5 Hz, 2H), 7.49 (d, J = 8.5 Hz, 2H), 7.31 (d, J = 16.0 Hz, 1H), 7.20 – 7.05 (m, 3H), 6.93 (d, J = 8.5 Hz, 2H), 3.86 (s, 3H)；
HRMS (ESI) calcd [M+H] + 389.9958, found 389.9940。
example 6 synthesis of compound If (r (n) =2, 4-dichloro):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (5.0g, 30mmol), heated to 105 deg.C for reaction, and the progress of the reaction was monitored by TLC, after about 2.5 h the reaction was complete. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (7.3g, 100 mmol), 2, 4-dichlorobenzaldehyde (5.2g, 30mmol) and sodium hydroxide (1.6g, 40mmol) and reacted at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), finishing the reaction for about 5 hours, then pouring the reaction liquid into ice water (185 mL), stirring, separating out a solid, filtering, and recrystallizing a filter cake by using a mixed liquid of ethanol and ethyl acetate in a volume ratio of 1:2 to obtain 2.4g of a yellow solid, namely (E) -2- (2, 4-dichlorostyryl) -5-bromo-4- (4-fluorophenyl) thiazole, wherein the yield is calculated to be 56.2%. m.p. 170-173 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.95 (dd, J = 9.0, 5.5 Hz, 2H), 7.66 (d, J = 16.5 Hz, 1H), 7.61 (d, J = 8.5 Hz, 1H), 7.45 (d, J = 2.5 Hz, 1H), 7.32 – 7.26 (m, 1H), 7.23 (d, J = 16.5 Hz, 1H), 7.17 (t, J = 9.0 Hz, 2H)；
HRMS (ESI) calcd C 17 H 9 BrCl 2 FNS [M+H] + 427.9073, found 427.9057。
example 7 synthesis of compound Ig (r (n) = o-methyl):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (2.5g, 15mmol), heated to 130 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 1 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (4.4g, 60 mmol), o-methylbenzaldehyde (2.4g, 20mmol) and sodium hydroxide (0.8 g, 20mmol) to react at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), ending the reaction for about 3 hours, pouring the reaction liquid into ice water (165 mL), stirring, extracting with ethyl acetate, desolventizing, and separating the residual liquid after desolventizing by using a mixed liquid of ethyl acetate and petroleum ether in a volume ratio of 1:3 through column chromatography to obtain 1.9 g of yellow solid, namely (E) -2- (2-methyl styryl) -5-bromo-4- (4-fluorophenyl) thiazole with the yield of 51.6%. m.p. 99-100 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.96 (dd, J = 9.0, 5.5 Hz, 2H), 7.68 – 7.55 (m, 2H), 7.31 – 7.21 (m, 3H), 7.20 – 7.11 (m, 3H), 2.47 (s, 3H)；
HRMS (ESI) calcd C 18 H 13 BrFNS [M+H] + 374.0009, found 373.9994。
example 8 synthesis of compound Ih (r (n) = m-methyl):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (6.6g, 40mmol), heated to 100 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 3h the reaction was complete. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (5.8g, 80 mmol), m-tolualdehyde (3.6g, 30mmol) and sodium hydroxide (2.0 g, 50mmol) to react at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), ending the reaction for about 5 hours, pouring the reaction liquid into ice water (175 mL), stirring, separating out a solid, filtering, and recrystallizing a filter cake by using a mixed liquid of n-hexane and ethyl acetate in a volume ratio of 1:2 to obtain 1.9 g of a yellow solid, namely (E) -2- (3-methyl styryl) -5-bromo-4- (4-fluorophenyl) thiazole with the yield of 54.6%. m.p. is 101-103 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.96 (dd, J = 8.5, 5.5 Hz, 2H), 7.41 – 7.27 (m, 4H), 7.24 (d, J = 16.5 Hz, 1H), 7.17 (t, J = 8.5 Hz, 3H), 2.40 (s, 3H)；
HRMS (ESI) calcd C 18 H 13 BrFNS [M+H] + 374.0009, found 373.9995。
example 9 synthesis of compound Ii (r (n) = p-methyl):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (2.5g, 15mmol), heated to 125 deg.C for reaction, and the progress of the reaction was monitored by TLC, after about 1.5 h, the reaction was complete. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (2.9g, 40mmol), p-tolualdehyde (2.4g, 20mmol) and sodium hydroxide (0.8 g, 20mmol) to react at room temperature. TLC detection reaction process, about 3.5h reaction is finished, then the reaction liquid is poured into ice water (185 mL), stirred, no solid is separated out, extracted and desolventized by ethyl acetate, the residual liquid after desolventization is separated by column chromatography by using mixed liquid of ethyl acetate and petroleum ether with the volume ratio of 1:5 to obtain 1.9 g yellow solid, (the product isE) -2- (4-methylstyryl) -5-bromo-4- (4-fluorophenyl) thiazole in 54.4% yield. 147-152 ℃ in m.p.;
1 H NMR (500 MHz, Chloroform-d) δ 7.95 (s, 2H), 7.44 (d, J = 7.0 Hz, 2H), 7.33 (d, J = 16.0 Hz, 1H), 7.26 – 7.10 (m, 5H), 2.39 (s, 3H)；
HRMS (ESI) calcd C 18 H 13 BrFNS [M+H] + 374.0009, found 373.9972。
example 10 synthesis of compound Ij (r (n) = m-bromo):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (5.8g, 35mmol), heated to 110 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 2.5 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (5.1g, 70 mmol), m-bromobenzaldehyde (5.6g, 30mmol) and sodium hydroxide (1.6g, 40mmol) and reacted at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), ending the reaction for about 4.5 hours, pouring the reaction solution into ice water (165 mL), stirring, extracting with ethyl acetate, desolventizing, and separating the residual liquid after desolventizing by using a mixed solution of ethyl acetate and petroleum ether in a volume ratio of 1:8 through column chromatography to obtain 2g of yellow solid, namely (E) -2- (3-bromostyryl) -5-bromo-4- (4-fluorophenyl) thiazole, wherein the yield is 45.7%, and the m.p. is 160-162 ℃.
1 H NMR (500 MHz, Chloroform-d) δ 7.95 (dd, J = 8.5, 5.5 Hz, 2H), 7.69 (s, 1H), 7.46 (t, J = 9.0 Hz, 2H), 7.34 – 7.25 (m, 2H), 7.22 (d, J = 16.0 Hz, 1H), 7.17 (t, J = 8.5 Hz, 2H)；
HRMS (ESI) calcd C 17 H 10 Br 2 FNS [M+H] + 437.8958, found 437.8968。
Example 11 synthesis of compound Ik (r (n) = p-trifluoromethyl):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (5.0g, 30mmol), heated to 115 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 2 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (4.4g, 60 mmol), p-trifluoromethylbenzaldehyde (5.2g, 30mmol) and sodium hydroxide (2 g, 50mmol) and reacted at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), ending the reaction for about 3 hours, pouring the reaction liquid into ice water (170 mL), stirring, extracting with ethyl acetate, desolventizing, and separating the residual liquid after desolventizing by using a mixed liquid of ethyl acetate and petroleum ether in a volume ratio of 1:6 through column chromatography to obtain 2.6 g of yellow solid, namely (E) -2- (4- (trifluoromethyl) styryl) -5-bromo-4- (4-fluorophenyl) thiazole, wherein the yield is 60.8%, and the m.p. is 123-125 ℃.
1 H NMR (500 MHz, Chloroform-d) δ 7.96 (dd, J = 9.0, 5.5 Hz, 2H), 7.65 (dd, J =12.5, 3.5 Hz, 4H), 7.39 (d, J = 16.0 Hz, 1H), 7.30 (d, J = 16.0 Hz, 1H), 7.17 (t, J = 8.5 Hz, 2H)；
HRMS (ESI) calcd C 18 H 10 BrF 4 NS [M+H] + 427.9726, found 427.9716。
Example 12 synthesis of compound Il (r (n) = o-methoxy):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (3.3g, 20mmol), heated to 120 ℃ for reaction, and the progress of the reaction was checked by TLC, after about 1.5 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (2.9g, 40mmol), o-methoxybenzaldehyde (2.7g, 20mmol) and sodium hydroxide (0.8 g, 20mmol) and reacted at room temperature. And (2) detecting the reaction process by TLC (thin layer chromatography), after about 3h of reaction is finished, pouring the reaction solution into ice water (185 mL), stirring, extracting with ethyl acetate, performing desolvation, and performing column chromatography on the residual liquid obtained after desolvation by using a mixed solution of ethyl acetate and petroleum ether in a volume ratio of 1:8 to obtain 2.4g of yellow solid, namely (E) -2- (2-methoxystyryl) -5-bromo-4- (4-fluorophenyl) thiazole with the yield of 61.6% and m.p. of 111-113 ℃.
1 H NMR (500 MHz, Chloroform-d) δ 7.96 (dd, J = 8.5, 5.5 Hz, 2H), 7.66 (d, J = 16.5 Hz, 1H), 7.56 (dd, J = 7.5, 1.5 Hz, 1H), 7.41 – 7.31 (m, 2H), 7.16 (t, J = 8.5 Hz, 2H), 7.00 (t, J = 7.5 Hz, 1H), 6.94 (d, J = 8.5 Hz, 1H), 3.93 (s, 3H)；
HRMS (ESI) calcd C 18 H 13 BrFNOS [M+H] + 389.9958, found 389.9945。
Example 13 synthesis of compound Im (r (n) = p-tert-butyl):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (4.1g, 25mmol), heated to 110 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 2 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (3.3g, 45 mmol), p-tert-butylbenzaldehyde (4.1g, 25mmol) and sodium hydroxide (0.8 g, 20mmol) to react at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), ending the reaction for about 3.5h, pouring the reaction liquid into ice water (165 mL), stirring, separating out a solid, filtering, and recrystallizing a filter cake by using a mixed liquid of ethanol and n-hexane in a volume ratio of 1:1 to obtain 1.9 g of a yellow solid, namely (E) -2- (4-tert-butylvinyl) -5-bromo-4- (4-fluorophenyl) thiazole with the yield of 45.5%. m.p. is 98-99 ℃;
1 H NMR (500 MHz, Chloroform-d) δ 7.98 (dd, J = 9.0, 5.5 Hz, 2H), 7.50 (d, J = 8.5 Hz, 2H), 7.44 (d, J = 8.0 Hz, 2H), 7.35 (d, J = 16.5 Hz, 1H), 7.24 (d, J = 16.5 Hz, 1H), 7.17 (t, J = 8.5 Hz, 2H), 1.37 (s, 9H)；
HRMS (ESI) calcd C 21 H 19 BrFNS [M+H] + 416.0478, found 416.0468。
example 14 synthesis of compound In (r (n) = ortho-bromine):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (5.0g, 30mmol), heated to 100 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 2.5 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (4.4g, 60 mmol), o-bromobenzaldehyde (4.6g, 25mmol) and sodium hydroxide (1.6g, 40mmol) to react at room temperature. The reaction process is detected by TLC, about 4h is finished, then the reaction liquid is poured into ice water (160 mL), stirred, solid is separated out, filtered, and the filter cake is recrystallized by using a mixed liquid of ethanol, ethyl acetate and n-hexane in a volume ratio of 1:1:1 to obtain 1.8g of yellow solid, namely (E) -2- (2-bromostyryl) -5-bromo-4- (4-fluorophenyl) thiazole, with the yield of 40.9%. m.p. 137-139 ℃;
1 H NMR (500 MHz, Chloroform-d) ä 7.96 (dd, J = 9.0, 5.5 Hz, 2H), 7.71 (d, J = 16.0 Hz, 1H), 7.68 (dd, J = 8.0, 1.5 Hz, 1H), 7.63 (dd, J = 8.0, 1.5 Hz, 1H), 7.35 (t, J = 7.0 Hz, 1H), 7.25 – 7.12 (m, 4H)；
HRMS (ESI) calcd C 17 H 10 Br 2 FNS [M+H] + 437.8957, found 437.8947。
example 15 synthesis of compound Io (r (n) = m-methoxy):
a100 mL three-necked flask was charged with 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole (3.7g, 10mmol), triethyl phosphite (6.6g, 40mmol), heated to 100 deg.C for reaction, and the progress of the reaction was checked by TLC, after about 3 h. Concentrating to remove redundant triethyl phosphite to obtain concentrated solution; to the resulting concentrated solution were added DMF (5.8g, 80 mmol), m-methoxybenzaldehyde (3.4g, 25mmol) and sodium hydroxide (1.8g, 45 mmol) and reacted at room temperature. And (3) detecting the reaction process by TLC (thin layer chromatography), ending the reaction for about 4.5 hours, pouring the reaction solution into ice water (170 mL), stirring, separating out a solid, filtering, recrystallizing a filter cake by using ethyl acetate to obtain 2.1 g of a yellow solid, namely (E) -2- (3-methoxystyryl) -5-bromo-4- (4-fluorophenyl) thiazole, wherein the yield is 54.5%, and m.p. is 167-168 ℃.
1 H NMR (500 MHz, Chloroform-d) δ 7.96 (dd, J = 8.5, 5.5 Hz, 2H), 7.64 (d, J = 16.0 Hz, 1H), 7.49 (d, J = 8.5 Hz, 1H), 7.26 – 7.08 (m, 5H), 6.79 (dd, J = 9.0, 3.0 Hz, 1H), 3.84 (s, 3H)；
HRMS (ESI) calcd C 18 H 13 BrFNOS [M+H] + 389.9958, found 389.9767。
Example 16 antitumor activity assay:
the stilbenes compounds containing thiazole structures prepared in examples 1-15 were labeled as compounds to be tested. Experiment the inhibitory activity of the test compound on topoisomerase I (TOP I) was tested using a DNA loose assay at a test concentration of 50. mu.M. The preparation process of 50 μ M test compound is: the test compound was dissolved in DMSO solvent to form a solution with a concentration of 50. mu.M.
The experimental method comprises the following steps: the enzyme used for the experiment is topoisomerase I (Topo I), and the experimental agent is stilbene compounds containing thiazole structures prepared in examples 1-15 (whether the test has an inhibiting effect on the topoisomerase I or not). If the compound has anti-tumor activity, a band in which supercoils can appear is particularly shown on an electrophoretogram.
The preparation method is as follows, and the reaction system volume of the following three preparations is 20. mu.L.
Blank DNA sample: mu.l of 0.5. mu.g/. mu.l negative supercoiled plasmid pBR322 DNA, 1. mu.l, buffer solution (20 mM Tris, pH 7.5, 0.1 mM EDTA, 10mM MgCl2, 100mM KCl,
50 μ g/mL acetylated BSA) to make up the volume to 20 μ L.
Sample of the test compound described above: mu.l of 0.5. mu.g/. mu.l negative supercoiled plasmid pBR322 DNA, 1. mu.l of 1.0. mu.g/. mu.l Top1 plasmid, 1. mu.l of 50. mu.M test compound, buffer make-up volume to 20. mu.l, buffer (20 mM Tris, pH 7.5, 0.1 mM EDTA, 10mM MgCl2, 100mM KCl,
50 μg/mL acetylated BSA)。
control positive drug (CPT) sample: mu.l of 0.5. mu.g/. mu.l negative supercoiled plasmid pBR322 DNA, 1. mu.l of 1.0. mu.g/. mu.l Top1 plasmid, 1. mu.l of 50. mu.M drug CPT, buffer make-up volume to 20. mu.l, buffer (20 mM Tris, pH 7.5, 0.1 mM EDTA, 10mM MgCl2, 100mM KCl,
50 μg/mL acetylated BSA)。
after the samples were prepared as described above, they were incubated at 37 ℃ for half an hour. After incubation, 4. mu.L of 6 Xloading buffer was added and mixed, and the mixture was applied to a sample well of 0.8% agarose gel and electrophoresed at 4.6V/cm for 1.5 hours. After the electrophoresis, the agarose gel was stained in a 1 Xgel red aqueous solution for 0.5 hour and photographed.
The inhibitory activity of the test compound was semi-quantified using the Fluor-S multimager gel imaging system based on the results of testing 50. mu.M test compound and 50. mu.M control positive drug CPT. The test results are shown in table 1.
The test results are shown in Table 1.
Antitumor Activity of Compounds Ia-Io at Table 150 μ M
Note a: TOP I (topoisomerase I) DNA Loose assay inhibitory Activity CPT (positive control drug having inhibitory activity against TOP I) was used as a control group. The data of the inhibitory activity of the obtained test compound relative to CPT are quantitatively expressed as follows: + + ++, representing more than 80% activity; + + + + represents between 40% and 79%; + means between 10% and 39%; +, less than 10% activity.
As shown by the DNA loosening experiment results in the table 1, the compound Ia-Io shown in the embodiment of the invention has certain inhibition activity on TOP I (DNA topoisomerase) at a concentration of 50 mu M, wherein the compound Ib has better inhibition activity on TOP I, and the inhibition activity of the compound Ib reaches about 70%.
The description is given for the sole purpose of illustrating the invention concept in its implementation form and the scope of the invention should not be considered as being limited to the particular form set forth in the examples.
Claims (8)
1. A Stilbene analogue containing a thiazole ring structure is characterized in that the structural formula is shown as a formula (I):
R (n) is o-chloro, p-chloro or 2, 4-dichloro;
the Stilbene analogue containing a thiazole ring structure is used for preparing an antitumor drug for inhibiting TOP I.
2. The method for synthesizing the Stilbene analogue containing the thiazole ring structure as claimed in claim 1, which comprises the following steps:
1) mixing 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole shown as a formula (II) and triethyl phosphite shown as a formula (III) in a molar ratio of 1: 1.5-5.0, then carrying out a reaction under a heating condition, and concentrating and removing redundant triethyl phosphite after TLC monitoring is carried out until the reaction is finished to obtain a concentrated solution;
2) adding a solvent DMF, sodium hydroxide and substituted benzaldehyde shown as a formula (IV) into the concentrated solution obtained in the step 1), reacting at room temperature, monitoring by TLC (thin layer chromatography) until the reaction is finished, and performing post-treatment on the reaction solution to obtain a Stilbene analogue containing a thiazole ring structure shown as a formula (I);
in the formula (IV), R (n) Is o-chloro, p-chloro or 2, 4-dichloro.
3. The method for synthesizing the Stilbene analogue containing the thiazole ring structure as claimed in claim 2, wherein the ratio of the amounts of the 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole shown in formula (II), the substituted benzaldehyde shown in formula (IV) and the sodium hydroxide is 1: 1.0-3.0: 1.0-5.0.
4. The method for synthesizing the Stilbene analogue with the thiazole ring structure as claimed in claim 2, wherein the molar ratio of the 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole shown in formula (II) to the DMF solvent is 1: 2.0-20.
5. The method for synthesizing the Stilbene analogue containing the thiazole ring structure as claimed in claim 4, wherein the molar ratio of 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole shown in formula (II) to DMF solvent is 1: 4.0-10.
6. The method for synthesizing Stilbene analogues containing thiazole ring structure as claimed in claim 2, wherein in step 1), the heating reaction temperature is 100-130 ℃, the heating reaction time is 1-3 hours, and in step 2), the reaction time at room temperature is 3-5 hours.
7. The method for synthesizing a Stilbene analogue containing a thiazole ring structure as claimed in claim 2, wherein in the step 2), the step of post-treating the reaction solution comprises: after the reaction is finished, adding a large amount of ice water into the reaction liquid, stirring, if solid is precipitated, filtering, and recrystallizing and purifying a filter cake by using an organic solvent to obtain the Stilbene analogue containing the thiazole ring structure shown in the formula (I); if no solid is precipitated, extracting with ethyl acetate, desolventizing, and separating the desolventized residual liquid by column chromatography to obtain the Stilbene analogue containing the thiazole ring structure shown in the formula (I); wherein the mass ratio of the added mass of the ice water to the 5-bromo-2- (bromomethyl) -4- (4-fluorophenyl) thiazole added in the step 1) is 33-50: 1.
8. The method for synthesizing the Stilbene analogue containing the thiazole ring structure as claimed in claim 7, wherein the organic solvent used for recrystallization is one or a mixture of two or more of ethanol, ethyl acetate and n-hexane; the eluent used for column chromatography separation is a mixed solution of ethyl acetate and petroleum ether in a volume ratio of 1: 3-8.
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| WO1994008982A1 (en) * | 1992-10-13 | 1994-04-28 | Nippon Soda Co., Ltd. | Oxazole and thiazole derivatives |
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| CN108863977A (en) * | 2018-06-19 | 2018-11-23 | 浙江工业大学 | The class diphenyl ethene compounds and its synthetic method of a kind of thiazole ring containing bromo and application |
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| WO1994008982A1 (en) * | 1992-10-13 | 1994-04-28 | Nippon Soda Co., Ltd. | Oxazole and thiazole derivatives |
| CN108605949A (en) * | 2018-06-19 | 2018-10-02 | 浙江工业大学 | A kind of application of the class diphenyl ethene compounds as fungicide containing thiazole ring |
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