Preparation method of hydroxytyrosin-d4
By performing deuterated and reduction reactions under a base catalyst and then hydrolyzing in an acidic environment, the high deuterated hydroxytyrosol-d4 was successfully synthesized, which solved the problem of lack of deuterated compound synthesis methods in the prior art, and achieved low Efficient preparation of cost, safety, environmental protection.
Patent Information
- Application Number
- CN202411349248.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-26
- Publication Date
- 2025-01-17
AI Technical Summary
There is a lack of effective methods in the prior art to synthesize deuterated compound hydroxytyrosol-d4 with special structures, especially when hydrogen-deuterium exchange reactions are carried out under mild conditions, and many sensitive functional groups are not suitable for harsh proton conditions.
Hydrogen-deuterium exchange reaction was carried out in the presence of a base catalyst. Pepper acetic acid was deuterated with the deuterium source, and then reduced to obtain pepper ethanol-d4, and hydrolyzed in an acidic environment to finally obtain hydroxytyrosol-d4. This method avoids the use of highly toxic agents and has low cost, mild reaction conditions and simple operation.
The synthesis of hydroxytyrosol-d4 with high deuteration (99%) was achieved, reducing costs, improving safety and environmental protection, and avoiding the use of highly toxic reagents. The reaction conditions are mild and the operation is simple.
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Figure CN119320311A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of deuterated compound synthesis, and in particular to a method for preparing hydroxytyrosol-d4. Background Art
[0002] Hydroxytyrosol is a natural polyphenol compound with strong antioxidant activity and multiple biological and pharmacological activities. It has antioxidant, anti-atherosclerotic, anti-thrombotic, antibacterial, anti-inflammatory and anti-tumor effects. Deuterated hydroxytyrosol has the same structure and similar properties as ordinary hydroxytyrosol. Because the carbon-deuterium bond has a shorter bond length and higher bond energy than the carbon-hydrogen bond, deuterated compounds have shown special application value in the fields of medicine, environmental analysis and detection, and development of new materials. At present, the demand for deuterated compounds with special structures is increasing, and it is imperative to develop new methods to synthesize deuterated compounds. Hydroxytyrosol-d4, a derivative obtained by isotope labeling of hydroxytyrosol, can be used as a tracer in the process of drug development as an isotope internal standard for mass spectrometry detection. In addition to reducing the matrix effect, the introduction of the isotope internal standard can also more accurately quantify the analyte. Its chemical structure is as follows: .
[0003] In the related art, there is a lack of reports on the preparation method of hydroxytyrosol-d4, so it is of practical significance to study the preparation method of hydroxytyrosol-d4. Deuterium labeling of organic compounds can be achieved by synthesis starting from suitable isotope-labeled precursors or directly by isotope exchange reactions, and the latter is more practical. However, the H / D exchange reactions of most acidic compounds reported in the literature involve quite harsh proton conditions, which are not friendly to many sensitive functional groups. Summary of the invention
[0004] The object of the present invention is to provide a method for preparing hydroxytyrosol-d4, which has the advantages of low cost, mild reaction conditions, simple operation process, simple post-treatment, greater safety and environmental protection, high yield, etc.
[0005] The technical solution adopted by the present invention to solve its technical problem is: A method for preparing hydroxytyrosol-d4 comprises the following steps: S10: In the presence of a base catalyst, piperone acetic acid and a deuterium source are subjected to hydrogen-deuterium exchange to obtain piperone acetic acid-d2; S20: reducing the piperoacetic acid-d2 to obtain piperoethanol-d4; S30: In an acidic environment, piperrol-d4 is hydrolyzed to obtain hydroxytyrosol-d4.
[0006] The reaction flow chart of the present invention is as follows: The key intermediate piperoacetic acid-d2 of the present invention uses CDCl3 as a deuterium source and solvent to undergo a simple and mild D / H exchange reaction with a high deuterium substitution rate, providing a new and efficient route for synthesizing hydroxytyrosol-d4.
[0007] As a preferred embodiment, in S10, the molar ratio of piperic acetic acid: base catalyst: deuterium source is 1:0.01-0.1:3-6.
[0008] As a preferred embodiment, in S10, the base catalyst is selected from at least one of NaOD, MeONa, Et3N, DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), and TBD (1,5,7-triazabicyclo[4.4.0]dec-5-ene); and the deuterium source is deuterated chloroform.
[0009] As a preferred embodiment, in S10, the hydrogen-deuterium exchange reaction is carried out at room temperature for 5-12 hours, and the reaction is quenched and separated and purified to obtain piperone acetic acid-d2.
[0010] As a preferred solution, S20 is specifically as follows: dissolving the reducing agent in an organic solvent, then adding an organic solvent solution of piperone acetic acid-d2, reacting for 1-2 hours, quenching the reaction, separating and purifying to obtain piperone ethanol-d4. The reaction temperature of S20 is controlled at -5~5°C.
[0011] As a preferred embodiment, the molar ratio of piperone acetic acid-d2: reducing agent is 1:1.1-2.0.
[0012] As a preferred embodiment, the reducing agent is selected from at least one of AlLiD4 and NaBD4; and the organic solvent is tetrahydrofuran.
[0013] As a preferred embodiment, in S30, the acidic environment is provided by an acid, or by a combination of an acid and a solid acid catalyst. The molar ratio of piperone acetic acid-d2 to the acid is 1:3.0-6.0. The optimal amount of the solid acid catalyst is one tenth of the mass of piperone acetic acid-d2.
[0014] As a preferred embodiment, the acid is selected from at least one of BBr3, HCl, HBr, HI, and HNO3; the solid acid catalyst is selected from at least one of Amberlyst 15, HND-32, and HND-34. Preferably, HCl is 36 wt% concentrated HCl, HBr is 48 wt% aqueous solution, HI is 55 wt% aqueous solution, and HNO3 is 68% concentrated HNO3.
[0015] As a preferred embodiment, in S30, the hydrolysis reaction is carried out under a protective atmosphere, the reaction temperature is controlled at -40°C to 25°C, the reaction time is 1-36 hours, and hydroxytyrosol-d4 is obtained after separation and purification.
[0016] The beneficial effects of the present invention are as follows: the present invention uses piperine acetic acid as the main raw material, and uses aprotic solvent deuterated chloroform as a deuterium source to obtain piperine ethanol-d4 with a high deuterium degree (99%) at room temperature through hydrogen / deuterium exchange reaction and reduction reaction with a high yield, and piperine ethanol-d4 is then subjected to catalytic hydrolysis reaction to obtain hydroxytyrosol-d4, and this step avoids the use of highly toxic reagents. The present invention has the characteristics of easy availability of raw materials, low price, lower cost, mild reaction conditions, simple operation process, avoidance of the use of highly toxic reagents, greater safety and environmental protection, and high yield. . BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 This is the NMR detection image of hydroxytyrosol-d4. DETAILED DESCRIPTION
[0018] The technical solution of the present invention is further described in detail below through specific embodiments.
[0019] In the present invention, unless otherwise specified, the raw materials and equipment used can be purchased from the market or are commonly used in the art. The methods in the following embodiments, unless otherwise specified, are all conventional methods in the art.
[0020] Example 1 S10: Preparation of piperoacetic acid-d2 1.80 g (10 mmol, 1.0 equiv.) of piperoacetic acid (SM) and 0.14 g (1 mmol, 0.1 equiv.) of TBD were added to 50 mL of deuterated chloroform, reacted at 25°C overnight, concentrated under reduced pressure, and then 50 mL of deuterated chloroform was added to the reaction bottle for the second exchange, and reacted at 25°C overnight. After the reaction, 14 mL of 1N HCl was used for quenching, washed twice with water, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain about 1.73 g of piperoacetic acid-d2, with a yield of 95.1% and an abundance of 99.4%.
[0021] S20: Preparation of pepper alcohol-d4 Add 15 mL of tetrahydrofuran to the reaction flask, cool to 0°C, add 0.47 g of AlLiD4 (12 mmol, 1.2equiv.), dissolve 1.82 g of piperidine-d2 (10 mmol, 1.0equiv.) in 15 mL of tetrahydrofuran, and add dropwise. After 1.5 hours of reaction, quench with 0.5 M hydrochloric acid, filter, and wash with ethyl acetate to obtain an organic phase, which is washed with water, dried, filtered, concentrated, and purified by column chromatography to obtain about 1.63 g of piperidine-d4, with a yield of 95.6% and an abundance of 99.4%.
[0022] S30: Preparation of Hydroxytyrosol-d4 Add 1.0 g (6 mmol, 1.0 equiv.) of pepper alcohol-d4, 2.4 g (24 mmol, 4.0 equiv.) of 36% HCl, and 10 mg of Amberlyst 15 to the reaction bottle, and react at room temperature for 36 hours under N2 protection. After the reaction is completed, filter, adjust the pH of the filtrate to 10 with sodium hydroxide solution in an ice-water bath, wash twice with DCM, adjust the pH of the aqueous layer to 1-2 with 3M HCl, extract three times with methyl tert-butyl ether, combine the methyl tert-butyl ether, dry over anhydrous sodium sulfate, decolorize with 5% activated carbon, and concentrate to obtain hydroxytyrosol-d4 ( 1 H NMR (400 MHz, CDCl3) δ 6.60-6.58 (m, 2H), 6.66-6.41 (m, 1H), 3.17(s, 1H)., Figure 1 , the same below) is about 0.89 g, the yield is 96.0%, and the abundance is 99.4%.
[0023] Example 2 S10: Preparation of piperoacetic acid-d2 Add 1.80 g (10 mmol, 1.0 equiv.) of piperoacetic acid (SM) and 0.04 g (1 mmol, 0.1 equiv.) of NaOD to 50 mL of deuterated chloroform, react at 25°C overnight, concentrate under reduced pressure, add 50 mL of deuterated chloroform to the reaction bottle, carry out the second exchange, react at 25°C overnight, after the reaction, quench with 14 mL of 1N HCl, wash twice with water, dry over anhydrous sodium sulfate, filter and concentrate to obtain about 1.55 g of piperoacetic acid-d2, with a yield of 85% and an abundance of 99.0%.
[0024] S20: Preparation of pepper alcohol-d4 Add 15 mL of tetrahydrofuran to the reaction flask, cool to 0°C, add 0.50 g of NaBD4 (12 mmol, 1.2 equiv.), dissolve 1.82 g of piperoacetic acid-d2 (10 mmol, 1.0 equiv.) in 15 mL of tetrahydrofuran, and add dropwise. After 1.5 hours of reaction, quench with 0.5 M hydrochloric acid, filter, and wash with ethyl acetate to obtain an organic phase, which is washed with water, dried, filtered, concentrated, and purified by column chromatography to obtain about 1.6 g of piperoethanol-d4, with a yield of 94.2% and an abundance of 99.0%.
[0025] S30: Preparation of Hydroxytyrosol-d4 To the reaction flask, add 1.0 g (6 mmol, 1.0 equiv.) of pepper alcohol-d4, 4.0 g (24mmol, 4.0 equiv.) of 48% HBr, and 10 mg of Amberlyst 15, and react at room temperature for 12 hours under N2 protection. After the reaction is completed, filter and adjust the pH of the filtrate to 10 with sodium hydroxide solution in an ice-water bath, wash twice with DCM, adjust the pH of the aqueous layer to 1-2 with 3M HCl, extract three times with methyl tert-butyl ether, combine the methyl tert-butyl ether, dry over anhydrous sodium sulfate, decolorize with 5% activated carbon, and concentrate to obtain about 0.79 g of hydroxytyrosol-d4, with a yield of 85.0% and an abundance of 99.0%.
[0026] Example 3 S10: Preparation of piperoacetic acid-d2 Add 1.80 g (10 mmol, 1.0 equiv.) of piperoacetic acid (SM) and 0.014 g (0.1 mmol, 0.01 equiv.) of TBD to 50 mL of deuterated chloroform, react at 25°C overnight, concentrate under reduced pressure, add 50 mL of deuterated chloroform to the reaction bottle, carry out the second exchange, react at 25°C overnight, after the reaction, quench with 14 mL of 1N HCl, wash twice with water, dry over anhydrous sodium sulfate, filter and concentrate to obtain about 1.64 g of piperoacetic acid-d2, with a yield of 90% and an abundance of 99.2%.
[0027] S20: Preparation of pepper alcohol-d4 Add 15 mL of tetrahydrofuran to the reaction flask, cool to 0°C, add 0.50 g of NaBD4 (12 mmol, 1.2 equiv.), dissolve 1.82 g of piperidine acetic acid-d2 (10 mmol, 1.0 equiv.) in 15 mL of tetrahydrofuran, and add dropwise. After 1.5 hours of reaction, quench with 0.5 M hydrochloric acid, filter, and wash with ethyl acetate to obtain an organic phase, which is washed with water, dried, filtered, concentrated, and purified by column chromatography to obtain about 1.6 g of piperidine ethanol-d4, with a yield of 94.2% and an abundance of 99.2%.
[0028] S30: Preparation of Hydroxytyrosol-d4 Add 1.0 g (6 mmol, 1.0 equiv.) of pepper alcohol-d4 to the reaction flask, dissolve it in 24 mL of dichloromethane, cool to -40°C, add 24 mL (24 mmol, 4.0 equiv.) of dichloromethane solution of BBr3 dropwise, react for 1 hour under N2 protection, and quench the reaction at -40°C. Separate the organic phase, wash twice with water, dry with anhydrous sodium sulfate, decolorize with 5% activated carbon, and concentrate to obtain about 0.84 g of hydroxytyrosol-d4, with a yield of 90.0% and an abundance of 99.2%.
[0029] Example 4 The difference from Example 1 is that: S10: the base catalyst for the preparation of piperoacetic acid-d2 is replaced by MeONa. The rest is the same as Example 1. The yield of hydroxytyrosol-d4 is 85%, and the abundance is 99.0%.
[0030] Example 5 The difference from Example 1 is that: S10: Preparation of piperoacetic acid-d2 The base catalyst is replaced by Et3N. Others are the same as Example 1. The yield of hydroxytyrosol-d4 is 70%, and the abundance is 99.0%.
[0031] Example 6 The difference from Example 1 is that: S10: the base catalyst for the preparation of piperoacetic acid-d2 is replaced by DBU. The rest is the same as Example 1. The yield of hydroxytyrosol-d4 is 82%, and the abundance is 99.1%.
[0032] Example 7 The difference from Example 1 is that: S30: the acid catalyst for the preparation of hydroxytyrosol-d4 is replaced by HND-32. Others are the same as Example 1. The yield of hydroxytyrosol-d4 is 92%, and the abundance is 99.3%.
[0033] Example 8 The difference from Example 1 is that: S30: the acid catalyst for the preparation of hydroxytyrosol-d4 is replaced by HND-34. Others are the same as Example 1. The yield of hydroxytyrosol-d4 is 90%, and the abundance is 99.3%.
[0034] Example 9 The difference from Example 1 is that: S30: the acid for preparing hydroxytyrosol-d4 is replaced by 68% concentrated HNO3 (18 mmol, 3.0 equiv.). Others are the same as Example 1. The yield of hydroxytyrosol-d4 is 83%, and the abundance is 99.0%.
[0035] Example 10 The difference from Example 1 is that: S30: the acid for the preparation of hydroxytyrosol-d4 is replaced by: 55% concentration of HI (36 mmol, 6.0 equiv.). Others are the same as Example 1. The yield of hydroxytyrosol-d4 is 92%, and the abundance is 99.2%.
[0036] The above-described embodiment is only a preferred solution of the present invention and does not limit the present invention in any form. There are other variations and modifications without exceeding the technical solution described in the claims.
Claims
1. A method for preparing hydroxytyrosol-d4, characterized in that, The following steps are involved: S10: In the presence of a base catalyst, piperone acetic acid and a deuterium source are subjected to hydrogen-deuterium exchange to obtain piperone acetic acid-d2; S20: reducing the piperoacetic acid-d2 to obtain piperoethanol-d4; S30: In an acidic environment, piperrol-d4 is hydrolyzed to obtain hydroxytyrosol-d4.
2. The preparation method according to claim 1, characterized in that: In S10, the molar ratio of piperic acetic acid: base catalyst: deuterium source is 1:0.01-0.1:3-6.
3. The preparation method according to claim 1, characterized in that: In S10, the base catalyst is selected from at least one of NaOD, MeONa, Et3N, DBU, and TBD; and the deuterium source is deuterated chloroform.
4. The preparation method according to claim 1, characterized in that: In S10, the hydrogen-deuterium exchange reaction is carried out at room temperature for 5-12 hours, and the reaction is quenched and separated and purified to obtain piperone acetic acid-d2.
5. The preparation method according to claim 1, characterized in that: S20 specifically comprises: dissolving the reducing agent in an organic solvent, then adding an organic solvent solution of piperone acetic acid-d2, reacting for 1-2 hours, quenching the reaction, and separating and purifying to obtain piperone ethanol-d4.
6. The preparation method according to claim 5, characterized in that: The molar ratio of piperone acetic acid-d2: reducing agent is 1:1.1-2.
0.
7. The preparation method according to claim 5, characterized in that: The reducing agent is selected from at least one of AlLiD4 and NaBD4; and the organic solvent is tetrahydrofuran.
8. The preparation method according to claim 1, characterized in that: In S30, the acidic environment is provided by an acid, or by a combination of an acid and a solid acid catalyst.
9. The preparation method according to claim 8, characterized in that: The acid is selected from at least one of BBr3, HCl, HBr, HI, and HNO3; the solid acid catalyst is selected from at least one of Amberlyst 15, HND-32, and HND-34.
10. The preparation method according to claim 1, characterized in that: In S30, the hydrolysis reaction is carried out under a protective atmosphere, the reaction temperature is controlled at -40°C to 25°C, the reaction time is 1-36 hours, and hydroxytyrosol-d4 is obtained after separation and purification.