A kind of cyanuric chloride oxime ester and its synthesis method

Abstract

The present invention provides a cyanuric chloride oxime ester and a synthesis method thereof, belonging to the field of organic synthesis. The cyanuric chloride oxime ester can be used as a DNA depolymerizing compound for depolymerizing DNA. The present invention also provides a simple and efficient method for synthesizing the cyanuric chloride oxime ester, using ketoxime as a starting material, reacting with cyanuric chloride and sodium carbonate to synthesize the target product in a single step. The entire synthesis method is simple to operate. The reaction process has mild reaction conditions, uses inexpensive raw materials, and has low production costs. In addition, the target product obtained by this process has a high yield and purity.

Description

Technical Field

[0001] This 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] Chemical DNA cleavage involves the intercalation of compounds into DNA, causing base breaks within the DNA to generate distinct DNA fragments and enable DNA sequence analysis. Due to the unique structure of small molecule compounds, they can precisely control the breakpoints by inducing DNA cleavage at specific locations, making them a research hotspot for DNA-related drugs. Currently reported DNA cleavage agents include guanidine isothiocyanate, Ru(II) bipyridine complexes, polypyridyl copper(I) complexes, and anthrone oxime ester compounds. Despite the numerous reports of these compounds, researchers remain uncertain about which structures are specifically capable of DNA cleavage. However, current research indicates that the structure, ring size, and geometry of small molecule compounds are crucial for intercalation into DNA. Furthermore, the complexity of their preparation processes and the cost of raw materials also significantly impact their industrial application. Therefore, the exploration of novel DNA cleavage compounds and their preparation processes is of great significance. Summary of the Invention

[0003] The present invention provides a tricyandiamide ester and a synthesis method thereof. The compound can be used to cleave DNA during ultraviolet light photolysis. The synthesis method provided by the present 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] .

[0006] The present invention provides a method for synthesizing the compound, comprising the following steps: adding a mixed solvent of water and methanol, sodium carbonate, and ketoxime to a reaction apparatus, adding cyanuric chloride in batches, and stirring to react to obtain the target compound I. The reaction formula is as follows: .

[0007] In some specific synthesis methods, the volume ratio of water to methanol is 1:4~8.

[0008] The reaction temperature is 10℃~30℃.

[0009] In some specific synthesis methods, the volume ratio of water to methanol is 1:5~7.

[0010] The reaction temperature is 10℃~20℃.

[0011] In some specific synthesis methods, cyanuric chloride is added in 10-20 portions.

[0012] Some specific synthesis methods include: after the reaction is completed, adding water, and then adding dropwise hydrochloric acid with a concentration of 10% to 40%, stirring, filtering, washing with water, and drying to obtain the target product I.

[0013] Some specific synthesis methods include using 1 to 5 L of cyanuric chloride water per mole, and 200 g to 1 kg of cyanuric chloride hydrochloric acid per mole, and stirring for 0.5 to 2 hours.

[0014] Some specific synthesis methods, the amount of water used per mole of cyanuric chloride is 3~4L, the number of washing times is 1~5 times, and the amount of water used for washing per mole of cyanuric chloride is 2~10L.

[0015] In a preferred embodiment, the molar ratio of the compound cyanuric chloride, ketoxime and anhydrous sodium carbonate is 1:1~5:1~5, preferably 1:3.2:3.5.

[0016] In a preferred embodiment, the volume ratio of water to methanol in the reaction solvent system is preferably 1:6.

[0017] Preferably, the reaction temperature is 15°C.

[0018] In a preferred embodiment, the post-treatment is as follows: after the reaction is completed, 2 L of water is added to the mixture per mol of cyanuric chloride, and dilute hydrochloric acid with a concentration of 37% per mol of cyanuric chloride is added dropwise at a rate of 400 g. The mixture is stirred for 0.5 to 1 hour, filtered, washed with clean water, and dried to obtain the target compound I.

[0019] The present invention provides a cyanuric chloride oxime ester that can be used as a DNA depolymerizing compound for depolymerizing DNA. The present invention also provides a simple and efficient method for synthesizing the cyanuric chloride oxime ester, using ketoxime as a starting material, and reacting it with cyanuric chloride and sodium carbonate in a single step to synthesize the target product. The entire synthesis method is simple to operate. The reaction process has mild reaction conditions, uses inexpensive raw materials, and has low production costs. In addition, the target product obtained by this process has a high yield and purity. DETAILED DESCRIPTION

[0020] 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 fall within the scope of protection claimed by the present invention.

[0021] The compound obtained in 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), and its chemical name is 10,10',10''-(1,3,5-triazine-2,4,6-triyl)tris(oxy))tris(azaneylylidene))tris(anthracen-9(10H)-one), and its structure is:

[0022]

[0023] 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 was plasmid pBR322 DNA.

[0024] Example 1:

[0025] To 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), and 9,10-dianthrone, 9-oxime (73.2g, 0.328mol, 0.328eq). Maintain stirring at 15°C for 30 minutes. Add cyanuric chloride in portions at 15°C, adding 1.5g every 10 minutes, for a total of 18.4g (0.1mol, 1eq). Stir continuously for 1 hour until the reaction solution becomes viscous. 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, and then washed twice with 500 mL of clean water, and dried in an oven at 80°C to obtain the product tricyandiamide (78 g, 95.87%). 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 Elemental analysis of the product using an organic element analyzer revealed the following: C, 72.56; H, 3.23; N, 11.27; O, 12.87. The product's ESI-MS (m / z) (M+) value was 744.37.

[0026] The method of using high-performance liquid chromatography to determine whether the reaction is complete is as follows: mobile phase: methanol-water (V:V = 3:1), with 0.15% mass fraction of anhydrous Na2SO4 and 1.6% phosphoric acid added; chromatographic column: octadecylsilane bonded silica gel as the filler (C18, 4.6 × 250 mm, 5μm); detection wavelength: 231 nm; flow rate: 1.0 mL / min; column temperature: 30℃; test solution: take one drop of sample into a 10mL volumetric flask, add methanol to the scale, sonicate for 10 minutes, filter, and take 5μl of the filtrate for injection). The reaction is considered complete when the HPLC result shows that the ketoxime is ≤10mV.

[0027] Example 2:

[0028] The synthesis method was similar to that of Example 1, except that the volume ratio of water to methanol was V:V = 1:1, and the reaction system was thicker. Finally, 77.2 g of tricyandiamide ester was obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was 8.5 mv.

[0029] Example 3:

[0030] The synthesis method was similar to that of Example 1, except that the volume ratio of water to methanol was V:V = 1:15, and 76 g of cyanuric acid oxime ester was finally obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was 9 mv.

[0031] Example 4:

[0032] The synthesis method was similar to that of Example 1, except that the volume ratio of water to methanol was V:V = 1:5, the solvent volume was increased to 800 mL, and 74.2 g of tricyandiamide ester was finally obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was 7.5 mv.

[0033] Example 5:

[0034] The synthesis method was similar to that of Example 1, except that the amount of anhydrous sodium carbonate was 40 g, 0.38 mol, 0.38 eq. Finally, 78 g of tricyandiamide ester was obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was 7.5 mv.

[0035] Example 6:

[0036] The synthesis method was similar to that of Example 1, except that the amount of anhydrous sodium carbonate was 50 g, 0.47 mol, 0.47 eq. Finally, 77.4 g of cyanuric acid chlorinated oxime ester was obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was 3 mv.

[0037] Example 7:

[0038] The synthesis method was similar to that of Example 1, except that the amount of anhydrous sodium carbonate was 33 g, 0.314 mol, 0.314 eq. Finally, 80 g of tricyandiamide ester was obtained. HPLC results showed that the ketone oxime response peak height of the reaction solution was much higher than 10 mv.

[0039] Example 8:

[0040] The synthesis method refers to Example 1, except that the amount of dianthrone oxime is 70 g, 0.31 mol, 0.31 eq. Finally, 72 g of tricyandiamide ester is obtained. HPLC results show that the ketone oxime response peak height of the reaction solution is 0.1 mv.

[0041] Example 9:

[0042] The synthesis method was similar to that of Example 1. The difference from Example 1 was that the amount of ketoxime was 85 g, 0.380 mol, 0.38 eq. Finally, 90 g of tricyandiamide ester was obtained. HPLC results showed that the ketoxime response peak height of the reaction solution was much higher than 10 mv.

[0043] Example 10:

[0044] The synthesis method was similar to that of Example 1, except that the reaction temperature was 0°C and the stirring time was extended to 3 h when cyanuric chloride was added. Finally, 78 g of cyanuric chloride oxime ester was obtained. HPLC results showed that the ketone oxime response peak of the reaction solution was 2 mv high.

[0045] Example 11:

[0046] 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.

[0047] Test example:

[0048] Compound I prepared in Example 1 was prepared into a 1 M stock solution using DMSO. A supercoiled DNA solution (closed: 50 μM / base pair) was then prepared using PBS buffer. The stock solution of Compound I was added to the DNA solution to a concentration of 10 μM. The DNA solution was irradiated with a UV lamp (312 nm, 16 W) at room temperature for 2 h. The cleavage products were then analyzed by gel chromatography. Analysis revealed that after 2 h of cleavage, the ratio of the open-ring relaxed form (Form II) to the closed supercoiled ring (Form I) reached 8:1.5.

[0049] The specific embodiments described above merely illustrate preferred implementations of the present invention and do not encompass the scope of protection of the present invention. Any modifications, improvements, and substitutions made without departing from the spirit of the present invention are within the scope of protection of the present invention.

Claims

1. Compounds of formula I: 。 2. The method for synthesizing the compound according to claim 1, wherein The following operations are included: A mixed solvent of water and methanol, sodium carbonate, and ketoxime were added to a reaction apparatus, and cyanuric chloride was added in batches. The mixture was stirred and reacted 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 to 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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