O-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing pyrazole structure and preparation method and application thereof
阅读说明：本技术 邻氯苯基取代含吡唑结构的螺[吲唑-吡唑啉]衍生物及其制备方法与应用 (O-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing pyrazole structure and preparation method and application thereof ) 是由 任小荣 罗蒙强 邓莉平 杜奎 李琰 席眉扬 吴春雷 沈润溥 于 2019-11-05 设计创作，主要内容包括：本发明属于医药技术领域,尤其涉及一种邻氯苯基取代含吡唑结构的螺[吲唑-吡唑啉]衍生物及其制备方法与应用。本发明一种如权利要求1所述的邻氯苯基取代含吡唑结构的螺[吲唑-吡唑啉]衍生物的制备方法,其特征在于,包括如下步骤：(1)1-苯基-3-甲基-5-(1,2,4-三唑基)-4-吡唑甲醛的合成；(2)1-苯基-3-甲基-5-(1,2,4-三氮唑-1-基)-4-吡唑甲醛腙的合成；(3)1-苯基-3-甲基-5-(1,2,4-三氮唑-1-基)-α-氯-4-吡唑甲醛腙的合成；(4)5-(2-氯亚苄基)-1-苯基-6,7-二氢-1<I>H</I>-吲唑-4(5<I>H</I>)-酮的合成；(5)邻氯苯基取代含吡唑结构的螺[吲唑-吡唑啉]衍生物的合成。(The invention belongs to the technical field of medicines, and particularly relates to o-chlorophenyl substituted spiro [ indazole-pyrazoline with a pyrazole structure]Derivatives, and preparation methods and applications thereof. The invention relates to o-chlorophenyl substituted spiro [ indazole-pyrazoline ] containing pyrazole structure according to claim 1]The preparation method of the derivative is characterized by comprising the following steps: (1) synthesis of 1-phenyl-3-methyl-5- (1,2, 4-triazolyl) -4-pyrazolecarboxaldehyde; (2) synthesizing 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone; (3) synthesizing 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde hydrazone; (4)5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1 H -indazole-4 (5) H ) -synthesis of ketones; (5) spiro [ indazole-pyrazoline with pyrazole structure substituted by o-chlorophenyl]Combinations of derivativesAnd (4) obtaining.)
1. An o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure is characterized in that the o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing the pyrazole structure has the following chemical structural formula:
wherein: Ar-2-ClC6H4-。
2. A method for producing an o-chlorophenyl-substituted spiro [ indazole-pyrazoline ] derivative having a pyrazole structure according to claim 1, which comprises the steps of:
(1) synthesis of 1-phenyl-3-methyl-5- (1,2, 4-triazolyl) -4-pyrazolecarboxaldehyde: dissolving 21mmol of 1,2, 4-triazole in 50mL of ethanol solvent, adding 25mmol of potassium hydroxide, performing ultrasound at normal temperature of 25 ℃ for half an hour, performing rotary evaporation on ethanol under reduced pressure, adding 40mL of DMSO, adding 15mmol of 5-chloro-1-phenyl-3-methyl-4-formylpyrazole in batches, performing ultrasonic treatment at 50 ℃ for 2 hours, pouring the mixture into 200mL of ice water after TLC detection is finished, precipitating light yellow solid, and performing suction filtration; drying the solid matter, and recrystallizing with ethanol-water to obtain a compound 1-phenyl-3-methyl-5- (1,2, 4-triazolyl) -4-pyrazole formaldehyde;
(2) synthesizing 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone: adding 22mmol of phenylhydrazine into a flask containing 10mL of tetrahydrofuran, refluxing and stirring in a boiling water bath until the phenylhydrazine is dissolved, slowly dropwise adding 20mL of anhydrous ethanol solution in which 20mmol of 1-phenyl-3-methyl-5- (1,2, 4-triazolyl) -4-pyrazolecarboxaldehyde is dissolved, continuing refluxing and stirring in the boiling water bath for 1h, and dropwise adding 10 drops of concentrated hydrochloric acid until a light yellow precipitate appears; refluxing and stirring in a continuous boiling water bath for 5 hours, stopping the water bath, adding 20mL of distilled water, stirring, deepening the color of a light yellow precipitate, and performing suction filtration to obtain a yellowish red needle-shaped product; washing with anhydrous ether for many times, and vacuum drying to obtain 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone;
(3) synthesis of 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde hydrazone: adding 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone into 50ml of 1, 2-dichloroethane and 30ml of isopropanol, completely dissolving the mixture by magnetic stirring, then adding tert-butyl hypochlorite into the mixture by three times in a cold salt bath at the temperature of-12 ℃, and violently stirring for 2 hours, wherein the temperature is controlled below-5 ℃ during the period to obtain light blue transparent liquid; transferring the product into a round-bottom flask, and evaporating the solvent at 50 ℃ under reduced pressure to obtain a light yellow oily substance; adding a small amount of petroleum ether, heating for mixing, dissolving, separating out a large amount of yellow solid, and performing vacuum filtration to obtain 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde hydrazone;
(4) synthesis of 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazol-4 (5H) -one: dissolving 1-phenyl-6, 7-dihydro-1H-indazole-4 (5H) -ketone 10mmol and 2-chlorobenzaldehyde 10mL in ethanol, adding 2mL of 40% NaOH aqueous solution, stirring at 80 ℃ for 3 hours, filtering and separating by using a Buchner funnel, washing a filter cake with water, recrystallizing and purifying by using ethanol, filtering and drying to obtain a product 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazole-4 (5H) -ketone;
(5) adding 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazole-4 (5H) -one and 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde hydrazone into 20mL of ethanol, dropwise adding a mixed solution of 0.5mL of pyridine and 10mL of ethanol into the solution by using a constant-pressure dropping funnel, continuously stirring at normal temperature for 2 hours after the dropwise adding is finished, tracking the reaction by TLC, filtering after the reaction is completed, concentrating the filtrate under reduced pressure, carrying out silica gel column chromatography, and eluting with ethyl acetate and petroleum ether to obtain a final product.
3. The method for producing an o-chlorophenyl-substituted spiro [ indazole-pyrazoline ] derivative having a pyrazole structure according to claim 2, wherein: the mass ratio of the 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone to the tert-butyl hypochlorite substance in the step (3) is 5: 8.
4. The method for producing an o-chlorophenyl-substituted spiro [ indazole-pyrazoline ] derivative having a pyrazole structure according to claim 2, wherein: in the step (4), the ratio of the amount of the 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde oxime to the amount of the 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazol-4 (5H) -one substance is 3: 2.
5. The o-chlorophenyl substituted spiro [ indazole-pyrazoline ] containing a pyrazole structure according to claim 2]A process for the preparation of derivatives, characterized in that: v in eluent in the step (5)(Ethyl acetate):V(Petroleum ether)＝1:8。
6. The use of the o-chlorophenyl substituted pyrazole-containing spiro [ indazole-pyrazoline ] derivative according to claim 1 in antitumor drugs.
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure, and a preparation method and application thereof.
Background
5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazol-4 (5H) -one, having the following chemical structure:
the 1, 3-dipolar cycloaddition reaction is the most important method for synthesizing five-membered heterocyclic compounds with good regioselectivity and body selectivity, and is also a more active reaction in heterocyclic pharmaceutical chemistry research. The triazole has aromaticity and abundant electrons in a molecular structure, can interact with enzymes and receptors in organisms by forming hydrogen bonds, and has various biological activities. The isoxazole skeleton is an important pharmacophore in the application of medicaments, and has remarkable physiological and pharmacological activities. In addition, spiroisoxazoles synthesized by 1, 3-dipolar cycloaddition of nitrile oxides to exocyclic double bonds have attracted attention by pharmacologists because they exhibit some important physiological properties. Therefore, the heterocyclic compound has high synthetic value in terms of pharmacology and synthesis angle.
Pyrazole derivatives have attracted considerable attention as a class of useful intermediates and as a variety of pharmaceutical activities that they themselves exhibit. In order to better study the influence of different heterocycles on the pharmacological activity caused by the aggregation in the same molecule, an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure is synthesized through 1, 3-dipolar cycloaddition reaction.
Disclosure of Invention
The invention aims to provide an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure and a preparation method thereof, and the o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing the pyrazole structure is prepared.
In order to achieve the purpose, the technical scheme of the invention is as follows:
an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure has the following chemical structural formula:
wherein: Ar-2-ClC6H4-。
A method for preparing the o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative having a pyrazole structure according to
(1) synthesis of 1-phenyl-3-methyl-5- (1,2, 4-triazolyl) -4-pyrazolecarboxaldehyde (compound 1): dissolving 21mmol of 1,2, 4-triazole in 50mL of ethanol solvent, adding 25mmol of potassium hydroxide, performing ultrasonic treatment at normal temperature of 25 ℃ for half an hour, performing rotary evaporation on ethanol under reduced pressure, adding 40mL of DMSO, adding 15mmol of 5-chloro-1-phenyl-3-methyl-4-formylpyrazole in batches, performing ultrasonic treatment at 50 ℃ for 2 hours, performing TLC detection, pouring the mixture into 200mL of ice water after the raw materials are reacted, separating out light yellow solid, and performing suction filtration; drying the solid matter, and recrystallizing with ethanol-water to obtain a compound 1-phenyl-3-methyl-5- (1,2, 4-triazolyl) -4-pyrazole formaldehyde (compound 1);
(2) synthesis of 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone (compound 2): adding 22mmol of phenylhydrazine into a flask containing 10mL of tetrahydrofuran, refluxing and stirring in a boiling water bath until the phenylhydrazine is dissolved, slowly dropwise adding 20mL of anhydrous ethanol solution in which 20mmol of 1-phenyl-3-methyl-5- (1,2, 4-triazolyl) -4-pyrazole formaldehyde (compound 1) is dissolved, continuing refluxing and stirring in the boiling water bath for 1h, and dropwise adding 10 drops of concentrated hydrochloric acid until a light yellow precipitate appears; refluxing and stirring in a continuous boiling water bath for 5 hours, stopping the water bath, adding 20mL of distilled water, stirring, deepening the color of a light yellow precipitate, and performing suction filtration to obtain a yellowish red needle-shaped product; washing with anhydrous ether for several times, and vacuum drying to obtain 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone (compound 2);
(3) synthesis of 1-phenyl-3-methyl-5- (1,2, 4-triazol-1-yl) - α -chloro-4-pyrazolecarboxaldehyde hydrazone (compound 3): adding 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone (compound 2) into 50ml of 1, 2-dichloroethane and 30ml of isopropanol, magnetically stirring to completely dissolve the mixture, then adding tert-butyl hypochlorite into a cold salt bath to-12 ℃, stirring vigorously for 2 hours, and during the period, controlling the temperature below-5 ℃ to obtain a light blue transparent liquid; transferring the product into a round-bottom flask, and evaporating the solvent at 50 ℃ under reduced pressure to obtain a light yellow oily substance; adding a small amount of petroleum ether, heating for mixing, dissolving, separating out a large amount of yellow solid, and performing vacuum filtration to obtain 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde hydrazone (compound 3);
(4) synthesis of 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazol-4 (5H) -one (Compound 4): dissolving 1-phenyl-6, 7-dihydro-1H-indazole-4 (5H) -one 10mmol and 2-chlorobenzaldehyde 10mL in ethanol, adding 2mL of 40% NaOH aqueous solution, stirring at 80 ℃ for 3 hours, filtering and separating by using a Buchner funnel, washing a filter cake with water, recrystallizing and purifying by using ethanol, filtering and drying to obtain a product 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazole-4 (5H) -one (compound 4);
(5) adding 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazole-4 (5H) -one (compound 4) and 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde hydrazone (compound 3) into 20mL of ethanol, dropwise adding a mixed solution of 0.5mL of pyridine and 10mL of ethanol into the solution by using a constant-pressure dropping funnel, continuously stirring at normal temperature for 2 hours after the dropwise adding is finished, carrying out TLC (thin layer chromatography) tracking reaction, filtering after the reaction is completed, concentrating the filtrate under reduced pressure, carrying out silica gel column chromatography, and eluting with ethyl acetate and petroleum ether to obtain a final product.
The mass ratio of the 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -4-pyrazole formaldehyde hydrazone (compound 2) to the tert-butyl hypochlorite substance is 5: 8.
The mass ratio of the 1-phenyl-3-methyl-5- (1,2, 4-triazole-1-yl) -alpha-chloro-4-pyrazole formaldehyde oxime (compound 3) to the 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazol-4 (5H) -one (compound 4) is 3: 2.
V in the eluent(Ethyl acetate):V(Petroleum ether)＝1:8。
The o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure is applied to antitumor drugs.
The invention provides an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure and a preparation method thereof, wherein the preparation method is characterized in that an isoxazole ring is introduced into a 5- (2-chlorobenzylidene) -1-phenyl-6, 7-dihydro-1H-indazole-4 (5H) -one structure by using a 1, 3-dipolar cycloaddition method, so that the spiro [ indazole-pyrazoline ] derivative containing the pyrazole substitution of the 5- (1-phenyl-3-methyl-4-1, 2, 4-triazolyl) structure is synthesized. The pyrazole-substituted spiro [ indazole-pyrazoline ] derivative containing a 5- (1-phenyl-3-methyl-4-1, 2, 4-triazolyl) structure, which is prepared by the invention, has a strong tumor cell inhibition effect, and provides a foundation for further application in the field of medicines.
Drawings
Fig. 1 is a flow chart of a preparation method of steps (1) to (3) in a preparation method of an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure according to the present invention;
fig. 2 is a flow chart of a preparation method in step (4) in the preparation method of an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure according to the present invention;
FIG. 3 is a flow chart of a preparation method of step (5) in the preparation method of an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure according to the present invention;
Detailed Description
The technical solutions of the present invention are further described in detail below with reference to the drawings and examples, which should not be construed as limiting the present invention.
Pyrazole derivatives have attracted considerable attention as a class of useful intermediates and as a variety of pharmaceutical activities that they themselves exhibit. In order to better study the influence of different heterocycles on the pharmacological activity caused by the aggregation in the same molecule, an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure is synthesized through 1, 3-dipolar cycloaddition reaction.
The invention provides an o-chlorophenyl substituted spiro [ indazole-pyrazoline ] derivative containing a pyrazole structure, which has the following chemical structural formula:
wherein: Ar-2-ClC6H4-。