CN106854223A - Mustargen quercetin derivative and its production and use - Google Patents

Abstract

The present invention relates to nitrogen mustard quercetin derivatives including their stereoisomers or tautomers and a preparation method thereof, a pharmaceutical composition containing such derivatives as active ingredients, and an application of the medicine in treating cancer . The preparation method of the derivatives firstly prepares the Mannich base of the quercetin by using the quercetin, formaldehyde and ethyl chloride, and then reacts with the substituted aminophosphoryl dichloride to prepare the nitrogen mustard quercetin derivatives. Nitrogen mustard derivatives of quercetin have antitumor effects.

Description

Nitrogen mustard quercetin derivative and its preparation method and use

technical field

The invention relates to nitrogen mustard quercetin derivatives and their application in pharmacy, belonging to the technical field of medicine.

Background technique

Quercetin is an important part of flavonoids, which has a wide range of pharmacological effects and biological activities, such as anti-oxidation and scavenging oxygen free radicals, lowering blood pressure, protecting myocardial ischemia, avoiding ischemia-reperfusion loss, enhancing immune function and Anticancer, antiviral and analgesic effects, etc. In recent years, the inhibitory effect of quercetin on the proliferation of human ovarian cancer, breast cancer, leukocytes, and gastrointestinal tumor cells has attracted much attention. Taking quercetin as the parent, modifying its structure and improving its drug activity is of great significance for the discovery of new drugs.

Nitrogen mustards are a class of important anti-tumor drugs that directly act on DNA. They have a broad anti-tumor spectrum and are widely used in clinical practice. But its disadvantage is that its metabolites are toxic. In order to reduce its toxic and side effects and further improve its anticancer activity, the inventors incorporated nitrogen mustard structural fragments into the quercetin structure to obtain a class of nitrogen mustard quercetin derivatives with anticancer activity.

Contents of the invention

The object of the present invention is to provide nitrogen mustard quercetin derivatives, including their stereoisomers or tautomers, which have anticancer effects.

Another object of the present invention is to provide the preparation method of the above nitrogen mustard quercetin derivatives, including their stereoisomers or tautomers.

Another object of the present invention is to provide the above nitrogen mustard quercetin derivatives, including the use of their stereoisomers or tautomers.

The present invention is described in detail below.

The invention provides a nitrogen mustard quercetin derivative, including its stereoisomers or tautomers, the structure of which is shown in the following formula:

In the formula: R is selected from the following substituents: H, saturated hydrocarbon group, unsaturated hydrocarbon group, aryl group, heterocyclic group, substituted aryl group, substituted heterocyclic group, substituted hydrocarbon group including chloroethyl, 1-carboxy (ester ) alkylalkyl, 2-hydroxyalkyl, arylmethyl, heterocyclylmethyl, arylethyl, heterocyclylethyl, substituted arylmethyl, substituted heterocyclylmethyl, substituted aryl Ethyl, heterocyclylethyl.

The structures of the nitrogen mustard quercetin derivatives, including their stereoisomers or tautomers, include:

The present invention also provides the preparation method of the nitrogen mustard quercetin derivative, including its stereoisomer or tautomer, comprising the following steps:

The nitrogen mustard quercetin derivatives of the present invention, including their stereoisomers or tautomers, have anticancer activity.

Beneficial effect: a nitrogen mustard quercetin derivative with anticancer activity is provided.

The invention is further illustrated by the following examples, but it should be noted that the scope of the invention is not limited in any way by these examples.

detailed description

Example 1

Preparation of Nitrogen Mustard Quercetin Derivatives (1)

Under the protection of _N2 , add 97.6mg (1.2mmol) of formaldehyde solution, 94.8mg (1.2mmol) of 2-chloroethylamine, 302mg (1.0mmol) of quercetin and 5 mL of ethanol into the closed reactor, add 4 drops Hydrochloric acid, heated to 75-80 ° C for 3 hours, cooled naturally, precipitated solid, filtered with suction, the crude product was separated by chromatography (V (n-butanol): V (water): V (HOAc) = 4:1:1, Eluted with methanol, 129 mg of quercetin Mannich base yellow solid was obtained, with a yield of 33.0%.

Take 393mg (1mmol) of the above-mentioned Mannich base, 431mg (2.2mmol) of 2-chloroethylphosphoryl dichloride, dissolve in 5mL of dry DMF, add dropwise a mixture of 0.6mL of _Et3N and 4mL of dry DMF under stirring at room temperature, After 30 minutes of dripping, react at room temperature for 2 hours, then heat to 60°C, TLC monitors that the reaction is complete, adjust the pH of the solution to 3-4 with 10% HCl solution, then add 50mL of water, filter to obtain the crude product, and pass through the chromatography plate Separation (V (n-butanol): V (water): V (HOAc) = 4:1:1, methanol elution, to obtain quercetin derivatives (1). MS (EIS) [M+H] ⁺ = 640.0.

Example 2

Preparation of nitrogen mustard derivatives of quercetin (2)

Replace 431mg (2.2mmol) 2-chloroethylphosphoryl dichloride in Example 1 with 462mg (2.2mmol) (N-2-chloroethyl-N'-methyl) phosphoryl dichloride, other operations are the same as The same as in Example 1, the quercetin derivative (2) was obtained with a yield of 72.5%. MS (EIS) [M+H] ⁺ = 668.0.

Example 3

Preparation of Nitrogen Mustard Quercetin Derivatives (3)

Replace 431mg (2.2mmol) of 2-chloroethylphosphoryl dichloride in Example 1 with 696mg (2.2mmol) L-(N-2-chloroethyl-N'-hydroxymethylbenzyl) phosphoryl dichloride Chlorine, other operations were the same as in Example 1 to obtain a quercetin derivative (3) with a yield of 71%. MS (EIS) [M+H] ⁺ = 880.1.

Example 4

Preparation of nitrogen mustard derivatives of quercetin (4)

Use 520mg (2mmol) phosphoryl mustard to replace 431mg (2.2mmol) 2-chloroethylphosphoryl dichloride in Example 1, and other operations are the same as in Example 1 to obtain quercetin derivatives (4), Yield 76%. MS (EIS) [M+H] ⁺ = 764.0.

Example 5

Preparation of Nitrogen Mustard Quercetin Derivatives (5)

Replace 431mg (2.2mmol) of 2-chloro in Example 1 with 565mg (2mmol) of L-[N-2-chloroethyl-N'-(2-carboxymethyl) ethyl] phosphoryl dichloride Ethamidophosphoryl dichloride, and other operations were the same as in Example 1 to obtain quercetin derivative (5) with a yield of 72%. MS (EIS) [M+H] ⁺ = 812.0.

Example 6

Preparation of Nitrogen Mustard Quercetin Derivatives (6)

Replace the 431mg (2.2mmol) in embodiment 1 with 597mg (2mmol) L-[N-2-chloroethyl-N'-(1-hydroxyl-2-carboxymethyl carboxyl) ethyl] phosphoryl dichloride ) 2-chloroethylphosphoryl dichloride, and other operations were the same as in Example 1 to obtain a quercetin derivative (6) with a yield of 67%. MS (EIS) [M+H] ⁺ = 842.1.

Example 7

Preparation of Nitrogen Mustard Quercetin Derivatives (7)

Replace 431mg (2.2mmol) of 2-chloroethylphosphoryl dichloride in Example 1 with 247mg (2mmol) (N-2-chloroethyl-N'-4-pyridyl) phosphoryl dichloride, other operations In the same manner as in Example 1, the quercetin derivative (7) was obtained with a yield of 69.1%. MS (EIS) [M+H] ⁺ = 794.0.

Example 8

Preparation of Nitrogen Mustard Quercetin Derivatives (8)

Replace 431mg (2.2mmol) of 2-chloroethylphosphoryl dichloride in Example 1 with 573mg (2mmol) (N-2-chloroethyl-N'-benzyl) phosphoryl dichloride, other operations and implementation Same as Example 1, the quercetin derivative (8) was obtained with a yield of 73.2%. MS (EIS) [M+H] ⁺ = 820.1.

Example 9

Study on Antitumor Activity of Nitrogen Mustard Quercetin Derivatives

one. Inhibition of Ascites S180 Tumor

Mice bearing S180 tumor cells that had grown well 7 days after inoculation were sacrificed by devertebralization, and the abdomen was disinfected with iodine on a clean workbench. After alcohol deiodination, the ascites tumor was extracted, and the ratio of 1:4 (ascites: normal saline) was mixed with normal saline. Dilute to cell suspension (cell density 2×10 ⁹ /L). The right axilla of each mouse was sterilized with iodine tincture and subcutaneously inoculated with 0.2 ml of the tumor cell fluid. On the second day after inoculation, the mice were randomly divided into negative control group, ifosfamide group, 5-fluorouracil group, quercetin positive control group and nitrogen mustard quercetin derivative group. All groups were administered orally for 10 consecutive days. The next day after the last administration, the mice were sacrificed by vertebral dissection, the tumor tissue was stripped, and the tumor weight and body weight (after tumor removal) were weighed. The experimental results were statistically processed to calculate the tumor inhibition rate (Table 1). The results showed that, compared with the positive control substance, the nitrogen mustard quercetin derivative had a significant inhibitory effect on the ascites-bearing S180 tumor mice. After treatment, the weight of the mice was comparable to that of the quercetin group, and was significantly higher than that of the ifosfamide group and the 5-fluorouracil group. The coat color was good, the tumor size was small, and the animals did not die, indicating that the nitrogen mustard derivatives of quercetin are toxic. smaller.

Tumor suppression rate (%)=(average tumor weight in control group-average tumor weight in treatment group)/average tumor weight in control group×100%

two. Inhibition of microvessel growth in S180 tumor tissue

On the 21st day of the above experiment, the mice were anesthetized and sacrificed, and the tumor tissues were excised, fixed in 40g/L formaldehyde solution, embedded in paraffin, and routinely sectioned with a thickness of 3-5µm. HE staining and immunohistochemical examination were performed respectively. SABC method was used for immunohistochemical examination. The primary antibody was rabbit anti-mouse CD31 antibody, which was replaced by PBS as a negative control, and the normal tissue adjacent to the tumor was used as a positive control. Counting method: select isolated yellow-brown vascular endothelial cells or cell clusters in the interstitial cells of tumor tissue, or those whose inner diameter of the vascular lumen is less than 8 red blood cells represent a single microvessel. The number of microvessels in 5 fields of view was counted under 200 times field of view (0.7386mm ³ ), and the average value was taken. Calculate the microvessel density (Table 2).

The results showed that the nitrogen mustard quercetin derivatives had a significant inhibitory effect on the growth of microvessels in the S180 tumor tissue.

Example 10

Inhibitory Effect of Nitrogen Mustard Quercetin Derivatives on TRAMP-C2 Prostate Cancer Cells

Take 77 C57BL/6 male mice, 6-8 weeks old, weighing 18-20g/mouse, after 2 days of adaptive feeding, inject TRAMP in the logarithmic growth phase subcutaneously on the right back of each mouse under sterile conditions -C2 single cell suspension 0.2mL, 5.0×106 cells, ² days after inoculation, were randomly divided into negative control group, quercetin control group, ifosfamide control group and nitrogen mustard quercetin derivative group. The negative control group was injected intraperitoneally with propylene glycol 0.1 mL/rat, 3 times a week, 10 times in 3 weeks in total. The quercetin control group, ifosfamide control group and nitrogen mustard quercetin derivative group were respectively dissolved in 0.1ml propylene glycol at 50 mg/kg body weight, injected intraperitoneally, 3 times a week, 10 times in 3 weeks in total. Pay attention to observe the spirit, diet and activity of the mice every day. Five weeks after the inoculation, the mice were killed by pulling the neck, the tumor was removed, the tumor weight was weighed, and the tumor inhibition rate was calculated (Table 3). The results showed that the nitrogen mustard quercetin derivatives had a significant inhibitory effect on TRAMP-C2 prostate cancer cells. The drug did not produce obvious toxic and side effects to mice within the dose range.

Claims (4)

1. mustargen quercetin derivative includes its stereoisomer or dynamic isomer, it is characterised in that general structure is as follows：

In formula：R is selected from substituents：H, saturated hydrocarbyl, unsaturated alkyl, aryl, heterocyclic radical, substitution aryl, substitution Heterocyclic radical, the alkyl of substitution include chloroethyl, 1- carboxylics（Ester）Base alkyl, 2- hydroxy alkylenes, arylmethyl, heterocyclic methyl, fragrant ethyl, Heterocycle ethyl, the arylmethyl of substitution, the heterocyclic methyl of substitution, fragrant ethyl, the heterocycle ethyl of substitution.

2. mustargen quercetin derivative according to claim 1, including its stereoisomer or dynamic isomer, its feature It is that instantiation includes following structural formula：

。

3. mustargen quercetin derivative according to claim 1, including its stereoisomer or dynamic isomer, its preparation Method is comprised the following steps：

。

4. the mustargen quercetin derivative according to right 1, including its stereoisomer or dynamic isomer, and such medicine Compositions, the application in terms for the treatment of cancer medicine.