CN107151243B - 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compounds, preparation method and biological activity evaluation thereof - Google Patents

2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compounds, preparation method and biological activity evaluation thereof Download PDF

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CN107151243B
CN107151243B CN201610941805.5A CN201610941805A CN107151243B CN 107151243 B CN107151243 B CN 107151243B CN 201610941805 A CN201610941805 A CN 201610941805A CN 107151243 B CN107151243 B CN 107151243B
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thiazole
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thiazoline
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CN107151243A (en
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唐丽娟
史大华
马桂珍
刘玮炜
王建
曹志凌
尹福军
陶传洲
赵跃强
刘蒙
王长智
朱荟龙
殷龙
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Jiangsu Ocean University
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    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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    • C07—ORGANIC CHEMISTRY
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    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
    • C07D277/20—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
    • C07D277/22—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 only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
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    • C07D277/20—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
    • C07D277/22—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 only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
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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 belongs to the technical field of thiazole derivative chemical synthesis and biological activity evaluation, and relates to a 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compound shown as a formula I. The invention also provides a method for preparing the 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole, a method for inhibiting urease activity and biological activity evaluation for inhibiting activities of Escherichia coli (Escherichia coli), Bacillus subtilis (Bacillus subtilis) and Staphylococcus aureus (Staphylococcus aureus). The 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compound has certain bioactivity, so the compound has better application prospects in the industries of medicines, pesticides and the like.
Figure DSA0000135728550000011
R＝3a：C6H5；3b：3‑CH3C6H4；3c：4‑FC6H4；3d：4‑ClC6H4；3e：4‑BrC6H4；3f：4‑CH3OC6H4；3g：3‑CH3OC6H4；3h：furanyl‑2‑yl；3i：thiophen‑2‑yl。

Description

2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compounds, preparation method and biological activity evaluation thereof
Technical Field
The invention relates to the technical field of organic synthesis, in particular to a 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compound, a preparation method and biological activity thereof.
Background
Marine organisms have been an important source of natural products and active natural products with novel structures, and particularly marine microorganisms have become an important marine medicine resource and are paid more and more attention. Among the marine natural products that have been found, the thiazole ring is the basic structural unit of many important physiologically active compounds, and pharmacological studies have shown that these natural products have a wide range of antitumor, antiviral (Shen, k., Peng, j.m., Li, w., Yan, x.w., Hu, a.x., chi.j.struct.chem.2014, 6, 855) antibacterial activities (Mislin, g.l., Burger, a., Abdallah, m.a., tetrahedron.2004, 60, 12139), and even exhibit properties of binding proteins, DNA, RNA (Akaji, k., Kiso, y., Total synthesis of thia. tetradron 1999, 55, 10685), etc., and these unique physiological activities are related to the thiazole ring structure (Ying y.c., Taori, k.k., k. 130.8455, soh.2008).
In addition to findings from nature, efforts have been made to study the total synthesis of natural products and their analogs and the synthesis of derivatives of unique structural fragments (Karegoudar, p., Karthikeyan, m.s., Prasad, d.j., Mahalinga, m.s., Holla, b.s., Kumari n.s., eur.j.med.chem.2008, 43, 261). Therefore, the method has certain significance for the synthesis of the compound containing the thiazole or the thiazoline unique fragment. Conventional synthesis of thiazoles or thiazolines is by reaction with sulfurizing reagents via α -halogenocopper, with the classical method being the Hantzsch method ((a) Hantzsch, a., Weber, j.h., Ber, dtsch, chem.ges.1887, 20, 3118(b) Aoyama, t., Murata, s., Arai, i., Araki, n., Takido, t., Suzuki, y, Kodomari, m., Tetrahedron, 2006, 14, 3201(c) Han, f.s., tokyama, h., Tohru, f.s., Chem commu, 2007, 3444(d) Busacca, c.a, Dong, y, Spinelli, e.m., m., teh, Tetrahedron, leitt, 17, lamban, lambo, t, t.35 (d), wooly, g.g., wolf, l.g., wolf, l.s.s.35, l.g., wolf, l.s.35, royal, l.g., wolf.g., wolf.s.s.s.s.s., l.s.35, wolf.g., wolf. The conventional method that ester is decarboxylated, acyl-chlorinated and then amidated is adopted in the synthesis route of the literature (Tang L.J., Qian B.H., Ma G.Z., Liu W.W., Chinese J Struc Chem, 2015, 4, 645-doped 647.), the invention improves the synthesis route to directly synthesize the target compound by amidation of ester and amine, simplifies the reaction route, obtains the environmental protection effect and is suitable for industrial production.
In conclusion, the invention takes natural marine metabolite Largozole as a template, carries out the design and synthesis of the heterocyclic compound containing the thiazole-thiazoline fusion skeleton, and carries out the biological activity test on the compound, and the compound with novel structure and certain biological activity is expected to be obtained.
Disclosure of Invention
The invention aims to provide a 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compound, a preparation method thereof, urease inhibiting activity and inhibiting effects on Escherichia coli (Escherichia coli), Bacillus subtilis (Bacillus subtilis) and Staphylococcus aureus (Staphylococcus aureus).
The technical scheme adopted by the invention is as follows:
the synthesis of 4-methyl-2-substituted-5- (2-thiazolinyl) thiazole compounds is characterized by comprising the following general formula:
Figure BSA0000135728570000021
in the formula
R＝3a：C6H5；3b：3-CH3C6H4；3c：4-FC6H4；3d：4-ClC6H4；3e：4-BrC6H4；
3f：4-CH3OC6H4；3g：3-CH3OC6H4；3h：furanyl-2-yl；3i：thiophen-2-yl
A preparation method of 2-substituted 4-methyl-5- (2-thiazolinyl) -1, 3 thiazole-compounds comprises the following steps:
A. nitrile compounds and 17 percent of amine sulfide aqueous solution react in an organic solvent, ice water is added into the reaction solution, and the thioamide compounds are obtained by crystallization
B. Refluxing the thioamide compound and 2-ethyl chloroacetoacetate in an ethanol solvent, tracking by TLC, removing the solvent under reduced pressure, and recrystallizing glacial acetic acid and water (V/V, 1: 3) to obtain the 2-substituted-4-methylthiazole-5-ethyl formate.
C. The 2-substituted-4-methylthiazole-5-ethyl formate reacts with ethanolamine to prepare N- (2-hydroxyethyl) -4-methyl-2-substituted thiazole-5-formamide.
D. The N- (2-hydroxyethyl) -4-methyl-2-substituted thiazole-5-formamide and phosphorus pentasulfide react in a toluene solvent in a ring-forming way without column chromatography, and dichloromethane and acetone are recrystallized to obtain the target compound.
The post-treatment method in the step B is glacial acetic acid and water recrystallization, and the product is purified by adopting simple recrystallization, so that the operation is simple, the experiment cost is reduced, and the method is suitable for industrialization.
In the step C, the 2-substituted-4-methylthiazole-5-ethyl formate and excessive ethanolamine are subjected to direct amidation reaction to prepare the N- (2-hydroxyethyl) -4-methyl-2-substituted thiazole-5-formamide, so that the acyl chlorination reaction is avoided, the method is environment-friendly, the operation is simple, and the production cost is reduced.
The purification step of the reaction in the step D is to recrystallize methylene dichloride and acetone (V/V, 4: 1) to replace column chromatography
The invention has the beneficial effects that:
1. the invention provides a synthesis method for synthesizing a thiazole-thiazoline compound by carrying out vulcanization, cyclization and amidation on a nitrile compound and then carrying out cyclization reaction, which avoids acyl chlorination reaction, and purifies the product by a recrystallization method through post treatment, and has the advantages of simple and convenient preparation reaction operation, high yield, environmental friendliness and suitability for industrial production.
2. The invention performs the biological activity evaluation of inhibiting the urease activity and inhibiting the activities of escherichia coli (Escherichia coli), Bacillus subtilis (Bacillus subtilis) and Staphylococcus aureus (Staphylococcus aureus) on the target compound, and has certain biological activity, thereby having better application prospects in the industries of medicines, pesticides and the like.
Drawings
The following is a brief description of the spectrum of the target compound
FIG. 1 is a hydrogen nuclear magnetic spectrum of 4-methyl-2-phenyl-5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
FIG. 2 is a diagram of the 4-methyl-2-phenyl-5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole carbon nuclear magnetic spectrum
FIG. 3 is a nuclear magnetic spectrum of 4-methyl-2- (3-methylphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
FIG. 4 is a diagram of the carbon nuclear magnetic spectrum of 4-methyl-2- (3-methylphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
FIG. 5 is a nuclear magnetic spectrum of 4-methyl-2- (4-fluorophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole for hydrogen production
FIG. 6 is a chart of 4-methyl-2- (4-fluorophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole prepared carbon nuclear magnetic spectrum
FIG. 7 shows the target product 4-methyl-2- (4-chlorophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole hydrogen nuclear magnetic spectrum
FIG. 8 is a carbon nuclear magnetic spectrum diagram of a target product 4-methyl-2- (4-chlorophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
FIG. 9 is a nuclear magnetic spectrum of 4-methyl-2- (4-bromophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazol
FIG. 10 is a nuclear magnetic spectrum of 4-methyl-2- (4-bromophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
FIG. 11 is a nuclear magnetic spectrum of 4-methyl-2- (4-methoxyphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
FIG. 12 is a carbon nuclear magnetic spectrum of 4-methyl-2- (4-methoxyphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
FIG. 13 is a nuclear magnetic spectrum of 4-methyl-2- (furan-2-yl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole hydrogen
FIG. 14 is a nuclear magnetic spectrum of 4-methyl-2- (furan-2-yl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole
Detailed Description
The first embodiment is as follows: preparation of aryl (heterocyclyl) thioamides
Under the condition of stirring at room temperature, 15mmol of aromatic nitrile and 20mL of anhydrous methanol are added into a dry round-bottom flask, 17% (27.5 mmol) of ammonium sulfide solution is dropwise added, the reaction solution is fully mixed and reacts at 55 ℃, and the progress of the reaction is monitored by TLC. After completion of the reaction, 100mL of ice water was slowly added to the reaction mixture to precipitate a yellow solid, which was then filtered off, washed with ice water and dried. The crude product can be recrystallized from ethanol and water.
Example two: preparation of 4-methyl-2-aryl (heterocyclyl) -1, 3-thiazole-5-carboxylic acid ethyl ester
8mmol of substituted thiobenzamide, 10mmol of ethyl 2-chloroacetoacetate and 30mL of absolute ethanol are stirred for reflux reaction, and the reaction progress is monitored by TLC. Crystallizing in ice water bath or concentrating under reduced pressure, and recrystallizing the residue with glacial acetic acid to obtain the target product. Dissolving 1g of crude product in 20ml of glacial acetic acid, slowly adding 50ml of ice water after complete dissolution, separating out white crystals, filtering and drying.
Example three: preparation of N- (2-hydroxyethyl) -4-methyl-2-aryl (heterocyclyl) -1, 3-thiazole-5-carboxamide
Reacting 2mmol of 4-methyl-2-aryl (heterocyclic) -1, 3-thiazole-5-ethyl formate and 15mL of ethanolamine at a certain temperature, monitoring the reaction process by TLC, adding 30mL (15mL multiplied by 2) of ethyl acetate and 20mL of water after the reaction is finished, extracting twice, drying an organic layer by anhydrous magnesium sulfate, and concentrating under reduced pressure to obtain the product.
[0042] Various temperatures were tried in the process, including 25 deg.C, 45 deg.C, 65 deg.C, 85 deg.C, 105 deg.C, and the progress of TLC monitoring is shown in Table 1.
TABLE 1 preparation times and yields at different temperatures using N- (2-hydroxyethyl) -4-methyl-2- (3-tolyl) -1, 3-thiazole-5-carboxamide as example
Figure BSA0000135728570000031
Example four: preparation of target product 2-substituted-4-methyl-5- (2-thiazolinyl) -1, 3-thiazole
Dissolve [0042] in 15mL of toluene]Adding 9.6mmol of triethylamine and P into the amide product2S54mmol, refluxing, TLC tracking, cooling to room temperature after the reaction is finished, pouring out the toluene layer, washing with water for 2-3 times, drying the organic phase with anhydrous sodium sulfate, concentrating, and recrystallizing the crude product with dichloromethane and acetone.
Example five: preparation of 4-methyl-2- (furan-2-yl) -1, 3-thiazole-5-carboxylic acid ethyl ester
Structural formula (I)
Figure BSA0000135728570000041
8mmol 2-furanthiobenzamide, 10mmol 2-chloroacetoacetic acid ethyl ester and 30mL absolute ethyl alcohol are stirred and refluxed for reaction, TLC monitors the reaction process, and the reaction system is placed in ice water bath for crystallization, and is filtered, filtered and dried. Dissolving 1g of crude product in 20ml of glacial acetic acid, slowly adding 60ml of ice water after complete dissolution, separating out white crystals, and performing suction filtration to obtain white needle-like crystals, wherein the yield is 95.8%, and m.p.66-67 ℃. IR (KBr，cm-1)v：3072.63，2025.91，1709.6，1618.32，1490.04，1318.07.1HNMR(CDCl3300 MHz): 7.58-6.58 (m, 3H, furane hydrogen), 4.38(q, 2H, J ═ 5.0Hz, ester methylene-COOCH2)，2.79(s，3H，thiazole-CH3)，1.41(t，3H，J＝5.0Hz，methyl，-O-CH2-CH3)
Example six: preparation of 4-methyl-2- (thien-2-yl) -1, 3-thiazole-5-carboxylic acid ethyl ester
Structural formula (I)
Figure BSA0000135728570000042
The preparation method is the same as that of the fifth embodiment, 2-thiophenecarboxamide is used for replacing 2-furanthiobenzamide, and the compound 4-methyl-2- (thiophene-2-yl) -1, 3-thiazole-5-ethyl formate is obtained. White solid, m.p.57-59 ℃. IR (KBr, cm)-1)v：3072.63， 2025.98，1709.85，1618.32，1541.13，1277.51.1H NMR(CDCl3300 MHz): 7.60-7.11 (m, 3H, thiophenehydrogen), 4.37(q, 2H, J ═ 5.0Hz, ester methylene-COOCH2)，2.77(s，3H，thiazole-CH3)，1.40(t，3H，J＝ 5.0Hz，methyl，-O-CH2-CH3).
Example seven: preparation of 4-methyl-2- (4-methoxyphenyl) -1, 3-thiazole-5-carboxylic acid ethyl ester
Structural formula (I)
Figure BSA0000135728570000043
The preparation method is the same as that of the fifth embodiment, 4-methoxy thiobenzamide is used for replacing 2-furan thiobenzamide, white needle-shaped crystals are obtained, the yield is 82.6%, and m.p.45-47 ℃. IR (KBr, cm)-1)v：2922.59，1716.00，1618.12，1490.04，1101.26.1H NMR(CDCl3300 MHz): 7.85-7.31 (m, 4H, phenyl ring hydrogen), 4.40(q, 2H, J ═ 5.0Hz, ester methylene-COOCH2)，2.83(s，3H，CH3O-Phenyl)，2.46(s，3H，thiazole-CH3)，1.43(t，3H，J＝5.0Hz，methyl， -O-CH2-CH3).
Example eight: preparation of 4-methyl-2- (4-bromophenyl) -1, 3-thiazole-5-carboxylic acid ethyl ester
Structural formula (I)
Figure BSA0000135728570000051
The preparation method is the same as that of the fifth example, 4-bromobenzene thiobenzamide is used for replacing 2-furan thiobenzamide, white solid is obtained, the yield is 80.5%, and the m.p.74-75 ℃ IR (KBr, cm)-1)v：2922.59，1716.00，1618.12，1396.74，1101.26.1H NMR (CDCl3300 MHz): 8.15-7.39 (m, 4H, benzenyl hydrogen), 4.48(q, 2H, J ═ 5.0Hz, ester methylene-COOCH2)， 2.89(s，3H，thiazole-CH3)，1.51(t，3H，J＝5.0Hz，methyl，-O-CH2-CH3).
EXAMPLE preparation of nonaN- (2-hydroxyethyl) -4-methyl-2- (furan-2-yl) -1, 3-thiazole-5-carboxamide
Structural formula (I)
Figure BSA0000135728570000052
4-methyl-2- (furan-2-yl) -1, 3-thiazole-5-ethyl formate 2mmol, ethanolamine 15mL and ethanol at 85 ℃ react for 50min, water 20mL is added for dilution, ethyl acetate (15mL × 2) is used for extraction twice, the organic phase is washed by water again, anhydrous magnesium sulfate is added for drying, and the product is obtained by decompression and concentration, wherein the yield is 72.8%, m.p.106-108 ℃ IR (KBr, cm, M.p.-1)v：3416.58， 3131.75，2940.59，1647.2，1541.29，1492.90，1369.75，1303.75，1283.85，1223.25。1H NMR (CD3OD, 300 MHz): 7.79-6.72 (m, 3H, furane hydrogen), 2.73(s, 3H, thiazole-CH)3) 3.80-3.39 (m, 2 methylene groups 4H, - -NH- -CH2-CH2-OH)
EXAMPLE preparation of decaN- (2-hydroxyethyl) -4-methyl-2- (thiophen-2-yl) -1, 3-thiazole-5-carboxamide
Structural formula (I)
Figure BSA0000135728570000053
The preparation is as in example nine, 4-methyl-2- (thiophen-2-yl) -1, 3-thiazole-5-carboxylic acid ethyl ester is used instead of 4-methyl-2- (furan-2-yl) -1, 3-thiazole-5-carboxylic acid ethyl ester. The product is a light yellow solid, the yield is 89.4 percent, and the m.p. is IR (KBr, cm) at the temperature of between 67 and 68 DEG C-1)v：3423.69，3131.75，2940.59，1647.2，1541.29，1492.90，1369.75，1303.75，1283.85， 1223.25。1H NMR(CDCl3300 MHz): 7.54-7.09 (m, 3H, thiophenehydrogen), 3.62-3.42 (dd, 2 methylene groups 4H, -NH-CH2-CH2-OH)，2.72(s，3H，thiazole-CH3)，2.03(s，1H，-OH)
EXAMPLE preparation of undec-N- (2-hydroxyethyl) -4-methyl-2- (3-tolyl) -1, 3-thiazole-5-carboxamide
Structural formula (I)
Figure BSA0000135728570000061
The procedure is as in example nine, 4-methyl-2- (4-tolyl) -1, 3-thiazole-5-carboxylic acid ethyl ester is used instead of 4-methyl-2- (furan-2-yl) -1, 3-thiazole-5-carboxylic acid ethyl ester. The product is a light yellow solid, the yield is 75 percent, and the m.p.141-142 ℃ IR (KBr, cm)-1)v：3431.43，3300.70，2928.23，2880.92，1632.41，1555.36，1413.66，1312.38。1H NMR (CDCl3300 MHz): 8.26(s, 1H, -NH-), 7.8-7.10 (m, 4H, benzene ring hydrogen), 3.66-3.46 (dd, 2 methylene groups 4H, -NH-CH)2-CH2-OH)，2.71(s，3H，thiazole-CH3)，2.40(s，3H，CH3-phenyl)1.75(s，1H，-OH)
EXAMPLE preparation of the twelve desired products 4-methyl-2-phenyl-5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3a)
Structural formula (I)
Figure BSA0000135728570000062
Dissolving 2mmol of N- (2-hydroxyethyl) -4-methyl-2-phenyl-1, 3-thiazole-5-formamide in 15mL of toluene, and adding 5mL of triethylamine and P2S55mmol, reflux reaction, TLC monitoring reaction, cooling to room temperature after reaction, decanting the upper toluene layer, washing with water 2 times, drying the organic phase over anhydrous sodium sulfate, concentrating under reduced pressure, and recrystallizing with dichloromethane and acetone (v/v, 5: 1) at 62.8% yield. White crystals, m.p.62-63 ℃. IR (KBr, cm)-1)v：2842.81，1620.29，1576.69，1430.45，1332.14。。1H NMR(CDCl3，300MHz)：7.95～7.43(m，5H，PhH)，4.39(t，2H，J＝6.0Hz，thiazoline ring N-CH2)，3.47(t，2H，J＝6.0Hz，thiazoline ring S-CH2)，2.76(s，3H，thiazole-CH3)。13C NMR(CDCl3，300MHz)：168.19(s，thiazole C，S-C＝N)，159.66(s，thiazoline C，S-C＝N)， 154.80(s，thiazole C，N-C)，133.10～126.45(m，phenyl C+thiazole 5)，64.17(s，thiazoline4，N-C)， 35.06(s，thiazoline 5)，21.49(s，C6H5-CH3)。(ES+)m/z：338(M+H+，100)
EXAMPLE thirteen preparation of the title product 4-methyl-2- (3-methylphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3b)
Structural formula (I)
Figure BSA0000135728570000063
The preparation method is the same as the twelve example, the yield is 48.8 percent, white crystals are obtained, and the temperature is m.p.68-69 ℃. IR (KBr, cm)-1)v：2848.28， 1600.14，1578.61，1458.74，1332.64。1H NMR(CDCl3500 MHz): 7.96-6.96 (m, 4H, benzene ring H), 4.41(t, 2H, J ═ 5.0Hz, thiazoline N-CH)2) 3.48(t, 2H, J ═ 5.0Hz, thiazoline S-CH)2) 2.78(s, 3H, thiazole CH)3)，2.43(s，3H，CH3Ph).13C NMR(CDCl3300 MHz): 168.44(S, thiazole, S-C ═ N), 159.63(S, thiazoline, S-C ═ N), 154.71(S, thiazole, N-C), 138.83-114.35 (m, benzene ring + thiazole C ═ C-S), 64.21(S, thiazoline N-C), 35.08(S, thiazoline C-C-S), 21.35(S, CH C-S)3Ph), 17.82(s, thiazole CH)3).(ES+)m/z：291(M+H+，100). (ES+)m/z：275(M+H+，100)
EXAMPLE preparation of the fourteen object product 4-methyl-2- (4-fluorophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3c)
Structural formula (I)
Figure BSA0000135728570000071
The preparation method is the same as that of the twelfth example, and the yield is 53.5 percent. White crystals, m.p.130-131 ℃. IR (KBr, cm)-1)v：2800， 1618.84，1596.42，1405.63，1213.7。1H NMR(CDCl3300 MHz): 7.94-7.10 (m, 4H, benzene ring H), 4.39(t, 2H, J ═ 6.0Hz, thiazoline N-CH)2) 3.46(t, 2H, J ═ 6.0Hz, thiazoline S-CH2) 2.75(s, 3H, thiazole CH)3).13C NMR(CDCl3300 MHz): 166.92(S, thiazole, S-C ═ N), 165.48(S, thiazoline, S-C ═ N), 162.98(S, benzene ring C-F), 159.57(S, thiazole, N-C), 129.70 to 116.03(m, benzene ring + thiazole C ═ C-S), 64.16(S, thiazoline N-C), 35.10(S, thiazoline C-S), 17.75(S, thiazole CH ═ N), and methods of making and using the same3).19F NMR(CDCl3，300MHz)：-109.33.(ES+) m/z：279(M+H+，100)
EXAMPLE fifteen preparation of the title product 4-methyl-2- (4-chlorophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3d)
Structural formula (I)
Figure BSA0000135728570000072
The preparation method is the same as the twelve example, the yield is 51.2 percent, white crystals are obtained, and the temperature is m.p.139-140 ℃. IR (KBr, cm)-1)v：2850.24，1617.94，1593.30，1398.36，1230.95。1H NMR(CDCl3，300MHz)：7.88～7.40(m，4H，Phenyl ring H), 4.39(t, 2H, J ═ 6.0Hz, thiazoline N-CH2) 3.48(t, 2H, J ═ 6.0Hz, thiazoline S-CH)2) 2.76(s, 3H, thiazole CH)3).13C NMR(CDCl3300 MHz): 166.72(S, thiazole, S-C ═ N), 159.53(S, thiazoline, S-C ═ N), 154.89(S, thiazole, N-C), 136.64 to 126.78(m, benzene ring + thiazole C ═ C-S), 64.18(S, thiazoline N-C), 35.12(S, thiazoline C-S), 17.77(S, thiazole CH ═ N), and methods of making and using the compounds3).(ES+)m/z：295(M+H+，100)
EXAMPLE sixteen preparation of the desired product 4-methyl-2- (4-bromophenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3e)
Structural formula (I)
Figure BSA0000135728570000081
The preparation method is the same as that of the twelfth embodiment, the yield is 40.5 percent, and the crystal is light yellow and has the temperature of m.p.136-137 ℃. IR (KBr, cm)-1)v： 2849.36，1617.94，1586.36，1497.79，1231.67。1H NMR(CDCl3500 MHz): 7.84-7.31 (m, 4H, benzene ring H), 4.42(t, 2H, J ═ 5.0Hz, thiazoline N-CH2) 3.51(t, 2H, J ═ 5.0Hz, thiazoline S-CH2) 2.80(s, 3H, thiazole CH)3).13C NMR(CDCl3300 MHz): 166.80(S, thiazole, S-C ═ N), 159.00(S, thiazoline, S-C ═ N), 154.94(S, thiazole, N-C), 133.23 to 125.01(m, benzene ring + thiazole C ═ C-S), 64.18(S, thiazoline N-C), 35.13(S, thiazoline C-S), 17.77(S, thiazole CH ═ N), and so on3).(ES+)m/z：338(M+H+，100)
EXAMPLE seventeen preparation of the desired product 4-methyl-2- (4-methoxyphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3f)
Structural formula (I)
Figure BSA0000135728570000082
The preparation method is the same as the twelve example, the yield is 42.6 percent, white crystals are obtained, and the temperature is m.p.133-134 ℃. IR (KBr, cm)-1)v：2849.24，1617.94，1586.36，1497.79，1231.67。1H NMR(CDCl3500 MHz): 8.06-7.72 (m, 4H, benzene ring H), 4.34(t, 2H, J ═ 5.0Hz, thiazoline N-CH2)，3.66(s，3H，CH3OPh), 3.46(t, 2H, J ═ 5.0Hz, thiazoline S-CH2) 2.75(s, 3H, thiazole CH)3).13C NMR(CDCl3300 MHz): 168.11(S, thiazole, S-C ═ N), 159.63(S, thiazoline, S-C ═ N), 154.65(S, thiazole, N-C), 128.23 to 114.35(m, benzene ring + thiazole C ═ C-S), 64.12(S, thiazoline N-C), 55.44(S, CH3OPh), 35.05(S, thiazoline C-S), 17.81(S, thiazole CH-S)3).(ES+) m/z：291(M+H+，100).
EXAMPLE preparation of the eighteenth object product 4-methyl-2- (3-methoxyphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3g)
Structural formula (I)
Figure BSA0000135728570000083
The preparation method is the same as that of the twelfth embodiment, the yield is 64.6 percent, white crystals are obtained, and the temperature is between m.p.80 and 81 ℃. IR (KBr, cm)-1)v：2850.62， 1677.32，1581.36，1437.81，1255.17。1H NMR(CDCl3500 MHz): 7.89-6.84 (m, 4H, benzene ring H), 4.41(t, 2H, J ═ 5.0Hz, thiazoline N-CH2)，3.90(s，3H，CH3OPh)3.49(t, 2H, J ═ 5.0Hz, thiazoline S-CH2) 2.78(s, 3H, thiazole CH)3).13C NMR(CDCl3300 MHz): 168.01(S, thiazole, S-C ═ N), 159.62(S, thiazoline, S-C ═ N), 154.72(S, thiazole, N — C), 134.33 to 111.15(m, benzene ring + thiazole C ═ C-S), 64.17(S, thiazoline N-C), 55.45(S, CH3OPh), 35.09(S, thiazoline C-S), 17.82(S, thiazole CH ═ S)3).(ES+)m/z：291(M+H+， 100).
EXAMPLE nineteen preparation of the title product 4-methyl-2- (furan-2-yl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3h)
Structural formula (I)
Figure BSA0000135728570000091
The preparation method is the same as that of the twelfth embodiment, the yield is 48.5 percent, and the crystal is brown and has the temperature of m.p.99-100 ℃. IR (KBr, cm)-1)v：3100.31，2848.28，1618.15，1582.27，1496.02，1258.62。1H NMR(CDCl3500 MHz): 7.75-6.37 (m, 3H, furan ring H), 4.42(t, 2H, J ═ 5.0Hz, thiazoline N-CH)2) 3.48(t, 2H, J ═ 5.0Hz, thiazoline S-CH)2) 2.72(s, 3H, thiazole CH)3).13C NMR(CDCl3300 MHz): 159.40(S, thiazoline, S-C ═ N), 157.90(S, thiazole, N-C), 154.67(S, thiazole, S-C ═ N), 148.48(S, furan carbon at 2 position), 144.30(S, furan carbon at 5 position), 125.72(S, thiazole C ═ C-S), 112.41(S, furan carbon at 3 position), 110.36(S, furan carbon at 4 position), 64.15(S, thiazoline N-C), 35.10(S, thiazoline C-S), 17.72(S, thiazole CH), and so on3).(ES+)m/z：291(M+H+，100).(ES+)m/z： 251(M+H+，100)
EXAMPLE twenty preparation of the target product 4-methyl-2- (thien-2-yl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3i)
Structural formula (I)
Figure BSA0000135728570000092
The preparation method is the same as that of the twelfth embodiment, the yield is 48.5 percent, and the crystal is brown and has the temperature of m.p.112-113 ℃. IR (KBr, cm)-1)v： 3100.31，2848.28，1620.30，1587.59，1461.93，131.16。1H NMR(CDCl3500 MHz): 7.72-6.93 (m, 3H, furan ring H), 4.40(t, 2H, J ═ 5.0Hz, thiazoline N-CH)2)3.49(t, 2H, J ═ 5.0Hz, thiazoline S-CH2) 2.75(s, 3H, thiazole CH)3).13C NMR(CDCl3300 MHz): 161.88(S, thiazoline, S-C ═ N), 159.44(S, thiazole, S-C ═ N), 154.50(S, thiazole, N-C), 136.81(S, thiophene carbon 2), 128.68 to 125.73(m, furan 3, 4, 5 carbon + thiazole C ═ C-S), 64.04(S, thiazoline N-C), 35.12(S,thiazoline C-C-S), 17.68(S, thiazole CH)3).(ES+) m/z：267(M+H+，100).
Determination of inhibitory Effect of target product on urease Activity
Performing primary screening and IC (integrated circuit) of target product on urease activity (1500, 20KU) by using sunrise full-automatic enzyme calibration instrument50And (4) measuring the value.
Enzyme solution
And (3) putting the weighed 24.000mg of urease into a brown bottle washed by the potassium permanganate solution, adding 6mL of phosphate buffer solution with the pH value of 6.8 by using a pipette gun, and uniformly shaking to fully dissolve the urease for later use.
Substrate solution
3.000g of urea and 2.000mg of phenol red are weighed in a volumetric flask, and the volumetric flask is filled with a buffer solution with the pH value of 6.8 to 100mL to prepare a substrate solution containing 0.002% of phenol red and 500mM of urea for later use.
Sample solution
0.025g of the sample was weighed on an analytical balance, added to a 25mL volumetric flask and made to volume with DMSO.
Design of experiments
Three parallel experiments were performed for each sample, with a blank of DMSO. After all the samples were added, incubation was carried out for half an hour.
Initial selection of urease inhibitors
After incubation for half an hour, the well enzyme label plate is put into a full-automatic enzyme label instrument and starts to be detected by a computer. Where the number of test cycles is set to eight to reduce the error. The results are shown in Table 2:
TABLE 2 inhibition of urease by the target products
Figure BSA0000135728570000101
The IC50 was determined for the inhibition rate of 50% or more.
4-methyl-2- (3-methylphenyl) -5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole (3b) was further diluted to a concentration of 1000ug/mL, 800ug/mL, 600ug/mL, 400ug/mL, 200ug/mL and IC50 was determined.
According to the curves of the sample concentration and the inhibition rate, the IC50 is found to be 729.93 ug/mL.
Determination of the bacteriostatic Activity
The strain is selected from Escherichia coli (Escherichia coli), Bacillus subtilis (Bacillus subtilis) and Staphylococcus aureus (Staphylococcus aureus)
The plate culture medium is beef extract peptone culture medium, and the liquid culture medium is beef extract peptone without agar culture medium.
Sample solution preparation [0074]
The strain is inoculated by a coating method after plate activation and liquid culture medium activation respectively.
The test of the bacteriostatic activity adopts an oxford cup, and the sample amount is 200 mul.
Streptomycin with a concentration of 1mg/ml is used as a positive control and the solvent DMSO is used as a blank control.
Each plate was replicated three times and placed in a bacterial incubator for 20 hours at 35 ℃. The activity was directly examined by the zone of inhibition, and the results are shown in Table 3.
TABLE 3 in vitro bacteriostatic Activity of target product pairs
Figure BSA0000135728570000111

Claims (4)

  1. The application of the 2-substituted-4-methyl-5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole compound is characterized in that the application is the application of the 2-substituted-4-methyl-5- (4, 5-dihydrothiazol-2-yl) -1, 3-thiazole compound as an effective component in preparing a urease inhibitor medicament; the 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compound has the following general formula:
    Figure DEST_PATH_IMAGE001
    r is 3-CH3C6H4Or furan-2-yl.
  2. 2. Use according to claim 1, characterized in that: the 2-substituted-4-methyl-5- (4, 5-dihydrothiazole-2-yl) -1, 3-thiazole compound is prepared by the following method: synthesizing a thiazole-thiazoline compound by carrying out cyclization reaction on substituted nitrile after sulfuration, cyclization and amidation, wherein the substituent is R; the method comprises the following specific steps:
    A. reacting substituted nitrile with 17% of amine sulfide aqueous solution in an organic solvent, adding ice water into the reaction solution, and crystallizing to obtain a thioamide compound;
    B. refluxing the prepared thioamide compound and 2-chloroacetoacetic acid ethyl ester in an ethanol solvent, tracking by TLC, removing the solvent under reduced pressure, and recrystallizing glacial acetic acid and water to obtain a 2-substituted-4-methyl-1, 3-thiazole-5-ethyl formate compound; the substituent is R;
    C. reacting 2-substituted-4-methyl-1, 3-thiazole-5-ethyl formate with ethanolamine to prepare N- (2-hydroxyethyl) -4-methyl-2-substituted thiazole-5-formamide;
    D. the N- (2-hydroxyethyl) -4-methyl-2-substituted thiazole-5-formamide and phosphorus pentasulfide react in a toluene solvent in a ring-forming way without column chromatography, and dichloromethane and acetone are recrystallized to obtain the target compound.
  3. 3. Use according to claim 2, characterized in that: in the step B: the recrystallization adopts a mixed solution of glacial acetic acid and water with the volume ratio of 1: 3.
  4. 4. Use according to claim 2, characterized in that: in the step D, a mixed solution of dichloromethane and acetone with the volume ratio of 4: 1 is adopted for recrystallization.
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Title
Synthesis, Crystal Strycture and Antibacterial Activities of 4-Methyl-2-(4-methylphenyl)-5-(2-thiazolinyl)-1,3-thiazole;Tang Li-Juan et al.;《Chinese J. Struct. Chem.》;20160430;第35卷(第4期);第645-650页；第646页Scheme1、第646-647页2.2部分、第649页Table 3以及右栏第1段、 *

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