The p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition is spread out
Biology and its preparation method and application
Technical field
The invention belongs to pharmaceutical technology fields, more particularly to one kind p-nitrophenyl substituted maleimide of N- containing pyrrazole structure
Amine α-terpinene cycloaddition derivative and its preparation method and application.
Background technique
N- p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition derivative chemical structural formula is as shown in Figure 1.
It is most important that 1,3- Dipolar Cycloaddition becomes synthesis five member ring heterocyclic compound with its good region and main selectivity
More active a kind of reaction in method and heterocyclic drug chemical research.
In recent years, due to the extensive bioactivity of chromone, anticancer, antibacterial, inhibit platelet aggregation etc. and by
Concern.So either still from a synthetic point of view from pharmacology, this heterocyclic compounds has very high synthesis to be worth.
Pyrazole derivatives are as a kind of useful intermediate and themselves shown a variety of pharmaceutical activity come out and by people
Extensive concern.
Study different heterocycles assemble in same molecule and on influence caused by pharmacological activity, it is anti-by dipole-diople interaction
Pyrazoles N- p-nitrophenyl maleimide α-terpinene cycloaddition derivative containing chromone structure should be synthesized, in drug
Research field has great importance.
Summary of the invention
The purpose of the present invention is to provide one kind p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinenes
Cycloaddition derivative and its preparation method and application, knot of the 6- fluoro chromone phenylhydrazone for thering is carboxaldehyde radicals to replace using 3 in pyrazoles
Replaced on structure, the maleimide α replaced to N- p-nitrophenyl-terpinene cycloaddition product carries out structure of modification.
To achieve the goals above, technical solution of the present invention is as follows:
One kind p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition derivative, it is described to contain pyrrole
Azoles structure N- p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition derivative chemical structural formula is as follows:
A kind of p-nitrophenyl substituted maleimide of N- containing pyrrazole structure amine α-terpinene cycloaddition derivative of the present invention
Preparation method, which is characterized in that the p-nitrophenyl substituted maleimide of N- containing the pyrrazole structure amine α-terpinene cycloaddition is spread out
The preparation method of biology, comprising:
Bicyclic [the 2,2,2] -5- octene -2,3- dicarboxylic anhydride of 1- isopropyl-4-methyl-and paranitroanilinum are dissolved separately in
In acetone solvent, p-nitrophenyl amine aqueous solution is added dropwise under stiring pungent containing bicyclic [2,2, the 2] -5- of 1- isopropyl-4-methyl -
The reaction flask of alkene -2,3- diacid anhydride solution, exothermic heat of reaction simultaneously gradually generate pale yellow precipitate, after room temperature reaction 1-2 hours, upper
It states and sequentially adds manganese acetate, triethylamine and aceticanhydride in reaction flask, reaction heating, precipitating is gradually dissolved, reacted at 50~60 DEG C
5-8 hours, solution became red-black by orange, is cooled to room temperature, precipitates through massive laundering, drying, produced with acetone recrystallization
Maleimide α-terpinene cycloaddition product that object N- p-nitrophenyl replaces;
Maleimide α-terpinene cycloaddition product and 6- fluoro chromone the phenylhydrazone mixing that N- p-nitrophenyl is replaced
In dehydrated alcohol, toluene-sodium-sulfonchloramide is added, flows back 12-15 hours, carries out addition reaction, with recrystallizing methanol, vacuum drying is contained
Pyrrazole structure N- p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition derivative.
Further, described by bicyclic [2,2,2] -5- octene -2, the 3- dicarboxylic anhydride (compound 1) of 1- isopropyl-4-methyl -
It is dissolved separately in acetone solvent with paranitroanilinum (compound 2), comprising:
Bicyclic [the 2,2,2] -5- of 1- isopropyl-4-methyl-that 2mmol is added in the acetone solvent of every 20~30mL is pungent
Alkene -2,3- dicarboxylic anhydride;
The paranitroanilinum of 2mmol is added in the acetone solvent of every 20~30mL.
Into a ground, maleimide α-terpinene cycloaddition product, the 6- fluoro color of the N- p-nitrophenyl substitution
The molar ratio of ketone phenylhydrazone and toluene-sodium-sulfonchloramide is 1:1:1-1.5.
It is further, described to sequentially add manganese acetate, triethylamine and aceticanhydride, comprising:
0.3-0.5 grams of manganese acetate is added in the acetone solvent of every 40~60mL;
15-20mL triethylamine is added in the acetone solvent of every 40~60mL;
30-35mL aceticanhydride is added in the acetone solvent of every 40~60mL.
Further, the maleimide α-terpinene cycloaddition product and 6- fluoro that N- p-nitrophenyl is replaced
Chromone phenylhydrazone is mixed in dehydrated alcohol, wherein is added what 1mmol N- p-nitrophenyl replaced in every 20-25mL dehydrated alcohol
Maleimide α-terpinene cycloaddition product and 1mmol 6- fluoro chromone phenylhydrazone.
The invention also provides one kind p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition
Application of the derivative in terms of preparing anti-tumor drug.
One kind N- containing pyrrazole structure p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition proposed by the present invention is spread out
Biology and its preparation method and application, since the maleimide α-terpinene cycloaddition product replaced in N- p-nitrophenyl draws
The structure for entering to have imported on the basis of five yuan of pyrazoles ring structures chromone can change pharmacological activity, and the present invention has formaldehyde using 3
The 6- fluoro chromone phenylhydrazone that base replaces is replaced in the structure of pyrazoles, the maleimide α-replaced to N- p-nitrophenyl
Terpinene cycloaddition product carries out structure of modification, the p-nitrophenyl substituted maleimide amine of N- containing the pyrrazole structure α-being prepared
Terpinene cycloaddition derivative for for Bel-7402 (human liver cancer cell), KB (cancer cell of oral cavity), (gastric cancer is thin by SGC7901
Born of the same parents), HO8901 (ovarian cancer cell), HL-60 (leukaemia cell), ECA109 (colon-cancer cell) all have stronger inhibition and live
Property, it provides the foundation for its developmental research for being used for potential drug.
Detailed description of the invention
Fig. 1 is N- p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition derivatives chemical structural formula;
Fig. 2 is the derivative materialization of present invention N- containing pyrrazole structure p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition
Learn structural formula;
Fig. 3 is present invention N- containing pyrrazole structure p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition derivative system
Preparation Method chemical formula schematic diagram.
Specific embodiment
Technical solution of the present invention is described in further details with reference to the accompanying drawings and examples, following embodiment is not constituted
Limitation of the invention.
It has been investigated that introducing five yuan of pyrroles in N- p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition derivative
The structure that chromone has been imported on the basis of azoles ring structure can change pharmacological activity.Experimental data is as follows:
Table 1
Based on the above experimental data, the 6- fluoro chromone phenylhydrazone for having carboxaldehyde radicals to replace using 3 in this application is in pyrazoles
Structure on replaced, to N- p-nitrophenyl replace maleimide α-terpinene cycloaddition product carry out structure change
It makes.
A kind of p-nitrophenyl substituted maleimide of N- containing pyrrazole structure amine α of the application-terpinene cycloaddition derivative
Chemical structural formula is as shown in Figure 2.
In one embodiment, one kind N- containing pyrrazole structure p-nitrophenyl substituted maleimide amine α-terpinene cycloaddition
The preparation method of derivative, comprising:
By bicyclic [the 2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) of 1- isopropyl-4-methyl-and paranitroanilinum
(compound 2) is dissolved separately in acetone solvent, is under stiring added to p-nitrophenyl amine aqueous solution drop (compound 2) different containing 1-
The reaction flask of propyl -4- methyl-bicyclo [2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) solution, exothermic heat of reaction and gradually
It generates pale yellow precipitate and sequentially adds manganese acetate, triethylamine and aceticanhydride in above-mentioned reaction flask, instead after room temperature reaction 1-2 hours
It should heat up, precipitating gradually dissolves, and reacts 5-8 hours at 50~60 DEG C, and solution becomes red-black by orange, is cooled to room temperature, sinks
It forms sediment through massive laundering, drying, obtains maleimide α-terpinene ring of product N- p-nitrophenyl substitution with acetone recrystallization
Addition product (compound 3);
The maleimide α that N- p-nitrophenyl is replaced-terpinene cycloaddition product (compound 3) and 6- fluoro chromone
Phenylhydrazone (compound 4) is mixed in dehydrated alcohol, and toluene-sodium-sulfonchloramide is added, and is flowed back 12-15 hours, is carried out addition reaction, is tied again with methanol
Crystalline substance, vacuum drying obtain the p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition derivative (chemical combination
Object 5).
Wherein, the chemical full name of compound 3 are as follows: (N- pairs of -5- octene -2,3- of 1- isopropyl-4-methyl-bicyclic [2,2,2]
Nitrobenzophenone) dicarboximide, in the present embodiment referred to as N- p-nitrophenyl replace maleimide α-terpinene ring
Addition product.
And the chemical full name of compound 5 are as follows: 3- (the fluoro- chromone -3- base of 6-) -1- phenyl -7- methyl -4- isopropyl-bicyclic
[2,2,2] octane simultaneously [2,3-d] 3aH, 7aH pyrazoles -2,3- (N- p-nitrophenyl) dicarboximide, in the present embodiment referred to as
For the p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition derivative.
Wherein, the synthesis of 6- fluoro chromone phenylhydrazone (compound 4), the 6- fluoro chromone that can there is carboxaldehyde radicals to replace with 3
The method of schiff bases is generated with phenylhydrazine dehydration to synthesize.
Embodiment 1 takes the phenylhydrazine of 2mmol to be added to fill in the flask of 10mL tetrahydrofuran, and boiling water bath return stirring is to molten
Then the ethanol solution into the 20mL 6- fluoro chromone for having carboxaldehyde radicals to replace dissolved with 2mmol 3 is slowly added dropwise in solution, continue
Boiling water bath return stirring 1 hour, 10 drop hydrochloric acid are added dropwise, there is pale yellow precipitate appearance.Continuous boiling water bath return stirring 5 hours, stop
Sealing bath, adds people's 20mL distilled water to stir, and pale yellow precipitate darkens, filter 6- fluoro chromone phenylhydrazone, yellowish red color are needle-shaped
Product.It is rinsed repeatedly with anhydrous ether, is dried in vacuo to obtain product 6- fluoro chromone phenylhydrazone (compound 4).
It should be noted that the present invention is not limited to the synthetic method of 6- fluoro chromone phenylhydrazone (compound 4), compound 4
Chemical name can also state are as follows: the fluoro- chromone -3- phenylhydrazone of 6-, which is not described herein again.
In one embodiment, by bicyclic [2,2,2] -5- octene -2, the 3- dicarboxylic anhydride (compound of 1- isopropyl-4-methyl -
1) it is dissolved separately in acetone solvent with paranitroanilinum (compound 2), comprising:
Bicyclic [the 2,2,2] -5- of 1- isopropyl-4-methyl-that 2mmol is added in the acetone solvent of every 20~30mL is pungent
Alkene -2,3- dicarboxylic anhydride;
The paranitroanilinum of 2mmol is added in the acetone solvent of every 20~30mL.
In one embodiment, maleimide α-terpinene cycloaddition product, 6- fluoro that N- p-nitrophenyl replaces
The molar ratio of chromone phenylhydrazone and toluene-sodium-sulfonchloramide is 1:1:1-1.5.
In one embodiment, manganese acetate, triethylamine and aceticanhydride are sequentially added, comprising:
0.3-0.5 grams of manganese acetate is added in the acetone solvent of every 40~60mL;
15-20mL triethylamine is added in the acetone solvent of every 40~60mL;
30-35mL aceticanhydride is added in the acetone solvent of every 40~60mL.
In one embodiment, the maleimide α-terpinene cycloaddition product and 6- fluorine N- p-nitrophenyl replaced
It is mixed in dehydrated alcohol for chromone phenylhydrazone, wherein 1mmol N- p-nitrophenyl is added in every 20-25mL dehydrated alcohol and replaces
Maleimide α-terpinene cycloaddition product and 1mmol 6- fluoro chromone phenylhydrazone.
Below by way of specific embodiment, replace Malaysia acyl to elaborate the application p-nitrophenyl of N- containing pyrrazole structure
Imines α-terpinene cycloaddition derivative (compound 5) preparation method:
Embodiment 2,
1), maleimide α-terpinene cycloaddition product synthesis that N- p-nitrophenyl replaces: 2mmol 1- is different
Propyl -4- methyl-bicyclo [2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) and 2mmol paranitroanilinum (compound 2)
It is dissolved separately in the acetone solvent of 20mL, p-nitrophenyl amine aqueous solution is added dropwise to containing 1- isopropyl-4-methyl-under stiring
The reaction flask of bicyclic [2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) solution, exothermic heat of reaction simultaneously gradually generate faint yellow sink
It forms sediment, after room temperature reaction 1 hour, in above-mentioned reaction flask, sequentially adds 0.3 gram of manganese acetate, 15mL triethylamine and 30mL aceticanhydride, instead
It should heat up, precipitating gradually dissolves, and reacts 5 hours at 50~60 DEG C, and solution becomes red-black by orange, is cooled to room temperature, precipitates
Through massive laundering, drying, added with maleimide α-terpinene ring that acetone recrystallization obtains the substitution of product N- p-nitrophenyl
At product (compound 3).
2), import pyrrazole structure: the maleimide α that 1mmol N- p-nitrophenyl is replaced-terpinene cycloaddition produces
Object (compound 3) and 1mmol 6- fluoro chromone phenylhydrazone (compound 4) are mixed in 20mL dehydrated alcohol, and 1.2mmol chlorine is added
Amine T flows back 12 hours, carries out addition reaction, and with recrystallizing methanol, vacuum drying obtains the p-nitrophenyl of N- containing pyrrazole structure and takes
For maleimide α-terpinene cycloaddition derivative (compound 5).
Embodiment 3,
1), maleimide α-terpinene cycloaddition product synthesis that N- p-nitrophenyl replaces: 2mmol 1- is different
Propyl -4- methyl-bicyclo [2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) and 2mmol paranitroanilinum (compound 2)
It is dissolved separately in the acetone solvent of 30mL, p-nitrophenyl amine aqueous solution is added dropwise to containing 1- isopropyl-4-methyl-under stiring
The reaction flask of bicyclic [2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) solution, exothermic heat of reaction simultaneously gradually generate faint yellow sink
It forms sediment, after room temperature reaction 2 hours, in above-mentioned reaction flask, sequentially adds 0.5 gram of manganese acetate, 20mL triethylamine and 35mL aceticanhydride, instead
It should heat up, precipitating gradually dissolves, and reacts 8 hours at 50~60 DEG C, and solution becomes red-black by orange, is cooled to room temperature, precipitates
Through massive laundering, drying, added with maleimide α-terpinene ring that acetone recrystallization obtains the substitution of product N- p-nitrophenyl
At product (compound 3).
2), import pyrrazole structure: the maleimide α that 1mmol N- p-nitrophenyl is replaced-terpinene cycloaddition produces
Object (compound 3) and 1mmol 6- fluoro chromone phenylhydrazone (compound 4) are mixed in 25mL dehydrated alcohol, and 1.5mmol chlorine is added
Amine T flows back 15 hours, carries out addition reaction, and with recrystallizing methanol, vacuum drying obtains the p-nitrophenyl of N- containing pyrrazole structure and takes
For maleimide α-terpinene cycloaddition derivative (compound 5).
Embodiment 4,
1), maleimide α-terpinene cycloaddition product synthesis that N- p-nitrophenyl replaces: 2mmol 1- is different
Propyl -4- methyl-bicyclo [2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) and 2mmol paranitroanilinum (compound 2)
It is dissolved separately in the acetone solvent of 25mL, p-nitrophenyl amine aqueous solution is added dropwise to containing 1- isopropyl-4-methyl-under stiring
The reaction flask of bicyclic [2,2,2] -5- octene -2,3- dicarboxylic anhydride (compound 1) solution, exothermic heat of reaction simultaneously gradually generate faint yellow sink
It forms sediment, after room temperature reaction 2 hours, in above-mentioned reaction flask, sequentially adds 0.4 gram of manganese acetate, 18mL triethylamine and 33mL aceticanhydride, instead
It should heat up, precipitating gradually dissolves, and reacts 6 hours at 50~60 DEG C, and solution becomes red-black by orange, is cooled to room temperature, precipitates
Through massive laundering, drying, added with maleimide α-terpinene ring that acetone recrystallization obtains the substitution of product N- p-nitrophenyl
At product (compound 3).
2), import pyrrazole structure: the maleimide α that 1mmol N- p-nitrophenyl is replaced-terpinene cycloaddition produces
Object (compound 3) and 1mmol 6- fluoro chromone phenylhydrazone (compound 4) are mixed in 25mL dehydrated alcohol, and 1.4mmol chlorine is added
Amine T flows back 13 hours, carries out addition reaction, and with recrystallizing methanol, vacuum drying obtains the p-nitrophenyl of N- containing pyrrazole structure and takes
For maleimide α-terpinene cycloaddition derivative (compound 5).
Fig. 3 shows the preparation process of the application compound 5, wherein 1 indicates that compound 1,2 indicates that compound 2,3 indicates
Compound 3,4 indicates that compound 4,5 indicates compound 5.
Experimental data is as follows:
The p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition derivative (compound 5) is light
Yellow crystal, yield 63.5%, m.p.192-193 DEG C.
1H NMR (DMSO) δ: 7.32-7.51 (m, 12H, Ar-H), 6.49 (s, 1H, C=C-H), 6.03 (d, J=
8.5Hz, 1H, H-5), 6.10 (1H, d, J=8.5Hz, 1H, H-6), 3.14 (1H, d, J=8.9Hz, 1H H-2), 2.86 (1H,
D, J=8.9Hz, 1H, H-3), 1.34 (1H, m, H-7a), 1.47 (1H, m, H-7b), 1.34 (1H, m, H-8a), 1.47 (1H, m,
), H-8b 2.56 (1H, m, H-9), 1.01 (3H, d, J=7.0Hz, H-10), 1.07 (3H, d, J=6.7Hz, H-11), 1.50
(3H,s,H-12).
IR 3457 (N-C=O), 3085 (ArH), 1723 (C=O), 1579 (C=N), 1298 (C-O-C) cm-1
M/e:634 (100.0%).
Anal.calcd.for C36H31FN4O6:C,68.13;H,4.92;N,8.83.
The p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition derivative (compound 5) is anti-
Tumor promotion measurement result:
Mtt assay measures the In-vitro Inhibitory Effect of 5 pairs of compound different tumor strains:
Compound 5 is dissolved with DMSO, dilute, tumour cell Bel-7402 (human liver cancer cell), KB (cancer cell of oral cavity),
SGC7901 (stomach cancer cell), HO8901 (ovarian cancer cell), HL-60 (leukaemia cell), ECA109 (colon-cancer cell) are in 96 holes
It is planted on plate into 4000/200 holes μ L/, 2 μ L of compound is added in every hole, final concentration of 12.0 μM, 6.0 μM, 3.0 μM, 1.5 μM, is total to
It is same as 37 DEG C, is incubated for 72 hours in 5%CO2 cell incubator, with DMSO (1%) for blank control.After 72 hours, it is added dense eventually
Degree is the MTT of 0.25mg/mL, is placed in 37 DEG C, 4 hours in 5%CO2 cell incubator, blots solvent later, and 100 μ are added in every hole
L DMSO is measured at 570nm absorbance (OD value) with enzyme linked immunological instrument, and the data obtained is for calculating IC50 value.Select inhibition
The high compound of activity measures the compound effects time difference under various concentration to the shadow in human tumor cells period and apoptosis
It rings.
The test-compound of various concentration carries out scalping with 96 orifice plates, according to resulting inhibiting rate, calculates IC50 value, as a result
It see the table below:
Table 2
Table 2 shows the p-nitrophenyl substituted maleimide amine of N- containing pyrrazole structure α-terpinene cycloaddition derivative and (changes
Close object 5) for Bel-7402 (human liver cancer cell), KB (cancer cell of oral cavity), SGC7901 (stomach cancer cell), HO8901 (oophoroma
Cell), HL-60 (leukaemia cell), ECA109 (colon-cancer cell) all have stronger inhibitory activity, be used for potential drug for it
Developmental research provide the foundation.
The above embodiments are merely illustrative of the technical solutions of the present invention rather than is limited, without departing substantially from essence of the invention
In the case where mind and its essence, those skilled in the art make various corresponding changes and change in accordance with the present invention
Shape, but these corresponding changes and modifications all should fall within the scope of protection of the appended claims of the present invention.