Cyanuric chloride oxime ester and synthesis method thereof

Abstract

The invention provides cyanuric chloride oxime ester and a synthesis method thereof, and belongs to the field of organic synthesis. The cyanuric chloride oxime ester can serve as a DNA depolymerization compound to be used for depolymerizing DNA, the invention further provides a simple and efficient method for synthesizing the cyanuric chloride oxime ester, ketoxime serves as a starting material and reacts with cyanuric chloride and sodium carbonate to synthesize a target product in one step, and the whole synthesis method is easy and convenient to operate. The reaction process has the advantages of mild reaction conditions, low price of used raw materials and low production cost. In addition, the target product prepared by the process has higher yield and purity.

Description

Technical Field

[0001] The present invention belongs to the field of organic synthesis, and specifically relates to a cyanuric chloride oxime ester and a synthesis method thereof. Background Art

[0002] DNA chemical cleavage is a method of embedding compounds into DNA to break the bases of DNA to obtain different DNA fragments and realize DNA sequence analysis. Since the unique structure of small molecule compounds can cause DNA to break at a specific position, thus accurately controlling the break point, it has become a research hotspot for DNA-related drugs. Currently reported DNA cleavage agents include guanidine isothiocyanate, Ru (II) bipyridine complexes, polypyridine copper (I) complexes, and anthrone oxime ester compounds. Although a large number of related compounds have been reported, researchers still cannot theoretically determine which structures can specifically cut DNA. However, through current research, it can be seen that the structure, ring size and geometry of small molecule compounds are crucial for embedding into DNA. At the same time, the complexity of the preparation process of these compounds and the price of raw materials also have an important impact on their industrial application. Therefore, it is of great significance to explore new structures of DNA cleavage compounds and their preparation processes. Summary of the invention

[0003] The invention provides a cyanuric chloride oxime ester and a synthesis method thereof, wherein the compound can be used to cut DNA during ultraviolet light photolysis. The synthesis method provided by the invention has mild reaction conditions, a safe and simple operation process, and the obtained target product has high purity and yield.

[0004] The specific technical scheme of the present invention is as follows: Compound of formula I: .

[0005] The present invention provides a method for synthesizing the compound, comprising the following operations: adding a mixed solvent of water and methanol, sodium carbonate, and ketoxime to a reaction device, adding cyanuric chloride in batches, and stirring to react to obtain the target compound I. The reaction formula is as follows: .

[0006] In some specific synthesis methods, the volume ratio of water to methanol is 1:4~8.

[0007] The reaction temperature is 10℃~30℃.

[0008] In some specific synthesis methods, the volume ratio of water to methanol is 1:5~7.

[0009] The reaction temperature is 10℃~20℃.

[0010] In some specific synthesis methods, cyanuric chloride is added in 10-20 portions.

[0011] Some specific synthesis methods, after the reaction is completed, add water, then dropwise add hydrochloric acid with a concentration of 10% to 40%, stir, filter, wash with water, and dry to obtain the target product I.

[0012] Some specific synthesis methods include using 1 to 5 L of cyanuric chloride water per mole, using 200 g to 1 kg of cyanuric chloride hydrochloric acid per mole, and stirring for 0.5 to 2 hours.

[0013] Some specific synthesis methods, the amount of water used for each mole of cyanuric chloride is 3~4L, the number of cleaning times is 1~5 times, and the amount of cleaning water used for each mole of cyanuric chloride is 2~10L of water.

[0014] In a preferred embodiment, the molar ratio of the compounds cyanuric chloride, ketoxime and anhydrous sodium carbonate is 1:1~5:1~5, preferably 1:3.2:3.5.

[0015] In a preferred embodiment, the volume ratio of water to methanol in the reaction solvent system is preferably 1:6.

[0016] Preferably, the reaction temperature is 15°C.

[0017] In the preferred embodiment, the post-treatment is as follows: after the reaction is completed, 2L of water is added to the mixture per mole of cyanuric chloride, and 400g of 37% dilute hydrochloric acid is added dropwise per mole of cyanuric chloride, and stirred for 0.5 to 1 hour. After filtering, the filter cake is washed with clean water and dried to obtain the target compound I.

[0018] The beneficial effects of the present invention are as follows: the present invention provides a cyanuric chloride oxime ester, which can be used as a DNA depolymerization compound for depolymerizing DNA. The present invention also provides a simple and efficient method for synthesizing cyanuric chloride oxime ester, using ketoxime as a starting material, and synthesizing a target product through a one-step reaction with cyanuric chloride and sodium carbonate, and the entire synthesis method is simple to operate. The reaction process has mild reaction conditions, the raw materials used are cheap, and the production cost is low. In addition, the target product obtained by the process has a high yield and purity. DETAILED DESCRIPTION

[0019] The present invention is further illustrated by the following examples. It should be correctly understood that the examples of the present invention are only used to illustrate the present invention, rather than to limit the present invention. Therefore, simple improvements to the present invention based on the method of the present invention all fall within the scope of protection claimed by the present invention.

[0020] The compound obtained by the present invention is named: 10,10',10''-(((1,3,5-triazine-2,4,6-triyl)tris(oxy))tris(azaneylylidene))tris(anthracen-9(10H)-one), the chemical name is 10,10',10''-(1,3,5-triazine-2,4,6-triyl)tris(oxy))tris(azaneylylidene))tris(anthracen-9(10H)-one), and the structure is:

[0021] In the following examples, various processes and methods not described in detail are conventional methods known in the art. The supercoiled DNA used in the test examples is plasmid pBR322 DNA.

[0022] Embodiment 1: In a 1000mL four-necked round-bottom glass flask, add 500mL of water and methanol (V:V=1:6), anhydrous sodium carbonate (36.5g, 0.34mol, 0.34eq), 9,10-dianthrone, 9-oxime (73.2g, 0.328mol, 0.328eq) and stir for 30min at 15℃. Add cyanuric chloride to the reaction flask in batches at 15℃, adding 1.5g every 10min, with a total of 18.4g (0.1mol, 1eq). After adding, continue stirring for 1h until the reaction solution becomes viscous, and add 1g of cyanuric chloride every hour, twice. After the reaction of the raw materials was completed, 200 mL of water was added, 37 g of 37% hydrochloric acid was added dropwise, the pH was adjusted to 6.2, stirred for 30 min, filtered, the filter cake was added with 400 mL of water and stirred for 30 min, filtered, washed twice with 500 mL of clean water, and dried in an oven at 80 ° C to obtain the product trimeric cyanuric chloride oxime ester (78 g, 95.87%). The HPLC results showed that the ketone oxime response peak height of the reaction solution was 0.5 mv. The molecular formula of the product is C 45 H 24 N6O6, the product was analyzed by organic element analyzer, and the analysis results were: C, 72.56; H, 3.23; N, 11.27; O, 12.87. The measured value of the product ESI-MS (m / z) (M+) was 744.37.

[0023] The method of using high performance liquid chromatography to determine whether the reaction is completed is as follows: mobile phase: methanol-water (V:V=3:1), and 0.15% mass fraction of anhydrous Na2SO4 and 1.6% phosphoric acid are added; chromatographic column: octadecylsilane bonded silica gel as filler (C18, 4.6 ×250 mm, 5μm); detection wavelength: 231 nm; flow rate: 1.0 mL / min; column temperature: 30 ℃; test solution: take 1 drop of sample into a 10mL volumetric flask, add methanol to the scale, ultrasonicate for 10 minutes, filter, and take 5μl of the filtrate for injection). The reaction is considered complete when the HPLC results show that ketoxime ≤10mv.

[0024] Embodiment 2: The synthesis method refers to Example 1, except that the volume ratio of water to methanol is V:V=1:1, the reaction system is thicker, and 77.2 g of cyanuric acid oxime ester is finally obtained. HPLC results show that the ketone oxime response peak height of the reaction solution is 8.5 mv.

[0025] Embodiment 3: The synthesis method refers to Example 1, except that the volume ratio of water to methanol is V:V=1:15, and 76 g of cyanuric chloride oxime ester is finally obtained. HPLC results show that the ketone oxime response peak height of the reaction solution is 9 mv.

[0026] Embodiment 4: The synthesis method refers to Example 1, except that the volume ratio of water to methanol is V:V=1:5, the solvent volume is increased to 800 mL, and finally 74.2 g of cyanuric acid oxime ester is obtained. HPLC results show that the ketone oxime response peak height of the reaction solution is 7.5 mv.

[0027] Embodiment 5: The synthesis method refers to Example 1, except that the amount of anhydrous sodium carbonate (40 g, 0.38 mol, 0.38 eq) was used, and 78 g of cyanuric chloride oxime ester was finally obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was 7.5 mv.

[0028] Embodiment 6: The synthesis method refers to Example 1, except that the amount of anhydrous sodium carbonate (50 g, 0.47 mol, 0.47 eq) was used, and 77.4 g of cyanuric chloride oxime ester was finally obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was 3 mv.

[0029] Embodiment 7: The synthesis method refers to Example 1, except that the amount of anhydrous sodium carbonate (33 g, 0.314 mol, 0.314 eq) was used, and 80 g of cyanuric chloride oxime ester was finally obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was much greater than 10 mv.

[0030] Embodiment 8: The synthesis method refers to Example 1, and the difference from Example 1 is that the amount of dianthrone oxime is (70 g, 0.31 mol, 0.31 eq), and finally 72 g of cyanuric chloride oxime ester is obtained. HPLC results show that the oxime response peak height of the reaction solution is 0.1 mv.

[0031] Embodiment 9: The synthesis method is the same as that of Example 1, except that the amount of ketoxime (85 g, 0.380 mol, 0.38 eq) is used, and 90 g of cyanuric chloride oxime ester is finally obtained. HPLC results show that the ketoxime response peak height of the reaction solution is much higher than 10 mv.

[0032] Embodiment 10: The synthesis method refers to Example 1, except that the reaction temperature is 0°C, and the stirring time is extended to 3h when cyanuric chloride is added, and finally 78g of cyanuric chloride oxime ester is obtained. HPLC results show that the ketone oxime response peak height of the reaction solution is 2mv.

[0033] Embodiment 11: The synthesis method refers to Example 1, except that the reaction temperature is 50°C, and 77.8 g of cyanuric chloride oxime ester is finally obtained. HPLC results show that the ketone oxime response peak height of the reaction solution is 1 mv.

[0034] Test example: The compound I prepared in Example 1 was prepared into a 1M mother solution with DMSO, and then a supercoiled DNA (closed: 50 μM / base pair) solution was prepared with a PBS buffer solution. The mother solution of compound I was added to the DNA solution to make the concentration of compound I 10 μM. The DNA solution was irradiated with an ultraviolet lamp (312 nm, 16 W) at room temperature for 2 h, and then the cleavage products were analyzed by gel chromatography. The analysis showed that the ratio of the open-ring relaxed form (Form II) to the closed supercoiled ring (Form I) reached 8:1.5 after 2 h of cleavage.

[0035] The specific embodiments described above only describe the preferred implementation methods of the present invention, but do not include the protection scope of the patent of the present invention. Any modification, improvement and substitution made without departing from the concept of the present invention shall fall within the protection scope of the present invention.

Claims

1. Compounds of formula I: 。 2. The method for synthesizing the compound according to claim 1, characterized in that: The following operations are included: A mixed solvent of water and methanol, sodium carbonate and ketoxime are added to a reaction device, and cyanuric chloride is added in batches, and the mixture is stirred to react to obtain the target compound I; .

3. The synthesis method according to claim 2, characterized in that The molar ratio of cyanuric chloride, ketoxime and anhydrous sodium carbonate is 1: 1-5: 1-5; The volume ratio of water to methanol is 1:1~15.

4. The synthesis method according to claim 3, characterized in that The reaction temperature is 0°C~80°C.

5. The synthesis method according to claim 4, characterized in that: The volume ratio of water to methanol is 1:4~8; The reaction temperature is 10℃~30℃.

6. The synthesis method according to claim 5, characterized in that: The volume ratio of water to methanol is 1:5~7; The reaction temperature is 10℃~20℃.

7. The synthesis method according to any one of claims 1 to 6, characterized in that After the reaction is completed, water is added, and then hydrochloric acid with a concentration of 10% to 40% is added dropwise, stirred, filtered, washed with water, and dried to obtain the target compound I.

8. The synthesis method according to claim 7, characterized in that For every mole of cyanuric chloride, the amount of water used is 1~5L, the amount of hydrochloric acid used is 200g~1kg, and stirring is carried out for 0.5~2 hours.

9. The synthesis method according to claim 8, characterized in that For every mole of cyanuric chloride, the amount of water used is 3~4L, the amount of washing water used is 2~10L, and the number of washing times is 1~5 times.

10. Use of the compound according to claim 1 in preparing a DNA cleavage agent.

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