Method for preparing crystal gingerenone a

Abstract

Disclosed in the present invention is a method for preparing a crystal gingerenone A, comprising the preparation of a single crystal and a polycrystal. The present invention is suitable for the technical field of the preparation of the compound. The crystals have a melting point of 79-81ºC and have E-trans-enone structural characteristics as revealed by X-ray crystallography. The specific method comprises: adding the compound in an oily form into an organic solvent at a weight at least twice that of the compound, heating the mixture until the compound is dissolved, then cooling the mixture to room temperature, and then allowing the mixture to stand at a constant temperature of 0-20ºC for crystallization. Subsequently, by means of filtration, washing, and drying, the crystal can be obtained. The single crystal thereof belongs to a monoclinic system and a space group P21. The present invention breaks through the current technical bottleneck of being unable to obtain a high-purity gingerenone A crystal form, and lays a foundation for the development and application thereof in the fields of food, medicines, cosmetics, etc.

Description

A method for preparing crystalline gingerone A Technical fields:

[0001] This invention belongs to the field of gingerone A crystal preparation technology, specifically relating to a gingerone A crystal and its preparation method. Background technology:

[0002] Gingerone A is a natural product extracted from plants, primarily those in the ginger family, such as ginger. Due to its unique bioactivity, this compound has attracted widespread attention in biomedical research in recent years. Research on gingererone A encompasses multiple fields, including traditional medicine, modern pharmacology, and food science.

[0003] In traditional medicine, plants of the ginger family have been used as herbal remedies for various ailments for thousands of years. Ginger, as a representative example, is widely used to relieve gastrointestinal discomfort, promote digestion, alleviate pain, and treat symptoms such as the common cold. These traditional uses have provided valuable clues for later scientific research.

[0004] With the development of modern pharmacology, scientists have begun to explore the pharmacological mechanisms of action of gingerone A at the molecular level. Studies have found that gingerone A possesses various biological activities, including anti-inflammatory, antioxidant, and anti-tumor effects, which are of great significance for the prevention and treatment of various chronic diseases. For example, in terms of anti-inflammation, gingerone A can reduce inflammatory responses by inhibiting the production and release of inflammatory mediators, thereby alleviating symptoms such as pain and swelling. It has high research and application value in the fields of medicine and health foods.

[0005] Currently, commercially available gingerone A is typically a pale yellow to golden yellow oily liquid with no fixed melting point. It is also prone to solidification and clumping after prolonged storage, making it difficult to process. It also has very high requirements for raw material storage conditions, which limits the application of gingerone A in food and pharmaceuticals. Therefore, there is an urgent need to research and develop methods to obtain gingerone A with higher purity or even a single crystal structure.

[0006] Summary of the Invention:

[0007] Therefore, the purpose of this invention is to provide a gingerone A crystal and a method for preparing the same.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] This invention provides a gingerone A crystal, including single crystals and polycrystalline forms, with E-trans-enone structural features ((E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hept-4-en-3-one), and a melting point of 79-81℃. The melting point of crystals obtained by different methods varies slightly, generally between 79-80℃.

[0010] This invention provides a single crystal of gingerone A, wherein the unit cell parameters of the single crystal of gingerone A are as follows:

[0011] Based on the above technical solution, the bond length parameters of the gingerone A single crystal are further as follows:

[0012] Based on the above technical solution, the bond angle parameters of the gingerone A single crystal are further as follows:

[0013] The present invention also provides a method for preparing the above-mentioned gingerone A crystals, comprising the following steps:

[0014] (1) Add gingerone A oil to an organic solvent of more than 2 times its weight, heat and dissolve to obtain a clear solution;

[0015] (2) Cool the clear solution obtained in step (1) to room temperature, keep it at a constant temperature and let it stand to allow the organic solvent to slowly evaporate and precipitate single crystals, or keep it at a constant temperature between 0 and -20℃ to precipitate crystals. After filtration, filter cake is obtained. Wash the filter cake 2-3 times with recrystallization solvent and dry it to obtain the final product.

[0016] Based on the above technical solution, further, the organic solvent mentioned in steps (1) and (2) is one or a mixture of two or more of the following solvents: dichloromethane, methanol, 95% ethanol, 95% methanol, diethyl ether, methyl tert-butyl ether, ethyl acetate, n-heptane, petroleum ether at 60-90℃, n-hexane, cyclohexane, and toluene.

[0017] Based on the above technical solution, further, in the organic solvents described in steps (1) and (2), the volume ratio of diethyl ether to methanol, ethanol, dichloromethane, ethyl acetate or methyl tert-butyl ether is 1:1 to 10:1.

[0018] Based on the above technical solution, further, in the organic solvents described in steps (1) and (2), the volume ratio of methyl tert-butyl ether to methanol, ethanol, dichloromethane or ethyl acetate is 5:1 to 10:1.

[0019] Based on the above technical solution, further, in the organic solvents described in steps (1) and (2), the volume ratio of n-heptane, n-hexane, cyclohexane or petroleum ether at 60-90℃ to dichloromethane, ethyl acetate, methyl tert-butyl ether or toluene is 2:1 to 5:1.

[0020] Based on the above technical solution, further, the gingerone A content in the gingerone A oil in step (1) is 95-98%.

[0021] Based on the above technical solution, further, the amount of organic solvent used in step (1) is 2 to 11 times the amount of gingerone A oil.

[0022] Based on the above technical solution, further, the temperature for dissolving the solution in step (1) is 30 to 70°C.

[0023] Based on the above technical solution, further, the cooling rate in step (2) is controlled at 0.1 to 2℃ / min.

[0024] Based on the above technical solution, further, the amount of recrystallization solvent used in step (2) is 2 to 10 times the amount of gingerone A oil.

[0025] Based on the above technical solution, further, the drying in step (2) includes conventional drying and vacuum drying. The temperature of conventional drying (including but not limited to forced air drying) is 30 to 60°C, the vacuum degree of vacuum drying is ≤ -0.085MPa, and the drying time is 5 to 24 hours.

[0026] This invention provides a method for preparing gingerone A single crystal, the preparation method specifically including the following steps:

[0027] (1) Add 0.2g of gingerone A oil with a purity of 95-98% to 0.5mL of dichloromethane and dissolve the gingerone A oil at 30℃ to obtain a clear solution.

[0028] (2) Cool the clear solution obtained in step (1) at a rate of 0.5℃ / min. After cooling to room temperature, keep it at a constant temperature and let it stand still to allow the solvent dichloromethane to slowly evaporate and precipitate single crystals. Filter the solution to obtain a filter cake, wash the filter cake with 0.5 mL of methyl tert-butyl ether, and dry it at 35℃ for 12 h to obtain gingerone A single crystal.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] 1. This invention provides a method for preparing gingerone A crystals. The resulting crystals have a measurable melting point, overcoming the current technical bottleneck that prevents those skilled in the art from obtaining gingerone A crystals with a definite melting point. The crystals can be used as a standard for the qualitative and quantitative analysis of this compound. Furthermore, the process route is clear, operable, and the yield can reach over 70%.

[0031] 2. The gingerone A single crystal prepared by this invention has a purity of over 99%, exists in a single crystal form, has high stability, a melting point of 79-81℃, and is not prone to clumping when stored for a long time. According to a search of the existing technology, no gingerone A single crystal of this invention has been disclosed. This invention provides important support for gingerone A as a raw material for food and pharmaceuticals. Attached image description:

[0032] To more clearly illustrate the embodiments of the present invention, the accompanying drawings involved in the embodiments will be briefly described below.

[0033] Figure 1 is a single crystal structure diagram of gingerone A single crystal prepared in Example 1.

[0034] Figure 2 shows the 1H NMR spectrum of the gingerone A single crystal prepared in Example 1.

[0035] Figure 3 shows the carbon NMR spectrum of the gingerone A single crystal prepared in Example 1.

[0036] Figure 4 is a microscope image of gingerone A crystals prepared in Example 2.

[0037] Figure 5 is a microscope image of gingerone A crystals prepared in Example 3. Detailed Implementation

[0038] The present invention will be described in detail below with reference to the embodiments. However, the implementation of the present invention is not limited thereto. Obviously, the embodiments described below are only some embodiments of the present invention. For those skilled in the art, other similar embodiments can be obtained without creative effort and all fall within the protection scope of the present invention.

[0039] In the examples, gingerone A oil was processed by Shanghai Medicilon Biopharmaceutical Co., Ltd.; the crystal diffraction experiment was performed by the laboratory of Dalian University of Technology; the high performance liquid chromatography (HPLC) detection conditions for gingerone A were as follows: column: Shim-pack Velox C18, 2.7 μm, 4.6*150 mm; column temperature: 30℃; mobile phase flow rate: 0.8 mL / min; mobile phase: A: 0.05% TFA aqueous solution, B: 0.05% TFA acetonitrile solution; elution gradient: 0 min 95% A, 1 min 95% A, 25 min 35% A, 28 min 10% A, 33 min 10% A, 33.1 min 95% A, 40 min 95% A, end; detection wavelength: 237 nm.

[0040] Example 1

[0041] This embodiment provides a method for preparing gingerone A single crystal, including the following steps:

[0042] (1) Add 0.2g of gingerone A oil with a purity of 95-98% to 0.5mL of dichloromethane and dissolve the gingerone A oil at 30℃ to obtain a clear solution.

[0043] (2) Cool the clear solution obtained in step (1) at a rate of 0.5℃ / min. After cooling to 20℃, keep it at a constant temperature to allow the solvent dichloromethane to slowly evaporate and crystals to precipitate. Filter the solution to obtain a filter cake, wash the filter cake with 0.5 mL of methyl tert-butyl ether, and vacuum dry it (35℃, -0.085 MPa) for 12 h to obtain the final product.

[0044] The yield of the obtained product was 76.3%, and the liquid phase purity of gingerone A was 99.52%.

[0045] The crystal structure of the obtained gingerone A crystal was determined by X-ray diffraction. The results are shown in Figure 1. The gingerone A crystal has E-trans-enone structure characteristics, is monoclinic, space group P21, and the cell parameters are shown in Table 1.

[0046] Table 1. Cell parameters of gingerone A

[0047] The specific bond length parameters of gingerone A crystals are shown in Table 2.

[0048] Table 2 Bond length parameters of gingerone A crystals

[0049] The specific parameters of the bond angles of gingerone A crystals are shown in Table 3.

[0050] Table 3 Bond angle parameters of gingerone A crystals

[0051] The proton NMR spectrum and carbon NMR spectrum of the prepared gingerone A single crystal are shown in Figure 2 and Figure 3, respectively.

[0052] Example 2

[0053] This embodiment provides a method for preparing gingerone A crystals, including the following steps:

[0054] 1) Add 0.2g of gingerone A oil with a purity of 95-98% to 0.8mL of methyl tert-butyl ether, and dissolve the gingerone A oil at 55℃ to obtain a clear solution;

[0055] 2) Cool the clear solution obtained in step (1) at a rate of 0.2℃ / min until it reaches room temperature. Keep it at a constant temperature and let it stand still. Then put it in a refrigerator at 0 to -4℃ and let it stand still at a constant temperature to precipitate crystals. Filter the solution to obtain a filter cake. Wash the filter cake with 1 mL of methyl tert-butyl ether and dry it at 60℃ for 8 hours to obtain crystals.

[0056] The yield of the obtained product was 75.0%, and the liquid phase purity of gingerone A was 98.80%. The crystal structure of the obtained product was determined by X-ray diffraction, and no single crystal structure was detected. The microscopic image of the obtained gingerone A crystal is shown in Figure 4.

[0057] Example 3

[0058] This embodiment provides a method for preparing gingerone A crystals, including the following steps:

[0059] 1) Add 0.2g of gingerone A oil with a purity of 95-98% to a mixed solvent of 1mL diethyl ether and 0.8mL methyl tert-butyl ether, and dissolve the gingerone A oil at 40℃ to obtain a clear solution;

[0060] 2) Cool the clear solution obtained in step (1) at a rate of 0.2℃ / min until it reaches room temperature. Keep it at a constant temperature and let it stand still. Then put it in a refrigerator at 0 to -20℃ and let it stand still at a constant temperature to precipitate crystals. Filter the solution to obtain a filter cake. Wash the filter cake with 1 mL of methyl tert-butyl ether and dry it at 50℃ for 12 hours to obtain crystals.

[0061] The yield of the obtained product was 80.7%, and the liquid phase purity of gingerone A was 98.47%. The crystal structure of the obtained product was determined by X-ray diffraction, and no single crystal was detected. The microscopic image of the obtained gingerone A crystal is shown in Figure 5.

[0062] Example 4

[0063] This embodiment examines the stability of the samples prepared in the above examples. The gingerone A single crystal sample prepared in Example 1 is designated as Sample 1, the gingerone A crystal prepared in Example 2 is designated as Sample 2, and the gingerone A crystal prepared in Example 3 is designated as Sample 3.

[0064] 1) High temperature test

[0065] The sample was placed in an open, clean container and kept at 60°C for 10 days. Samples were taken on the 5th and 10th days to test whether the liquid phase content and crystal form of the sample changed.

[0066] 2) High humidity test

[0067] The sample was placed in an open, clean container, then placed in a humidity-controlled, sealed container and kept at 25°C and 90% ± 5% relative humidity for 10 days. Samples were taken on the 5th and 10th days. After drying, the liquid phase content and crystal form of the sample were tested to determine if they had changed.

[0068] 3) Strong light irradiation test

[0069] The sample was placed in an open, clean container and then placed in a light box equipped with fluorescent lamps. It was kept for 10 days under an illuminance of 1000 lx ± 100 lx. Samples were taken on the 5th and 10th days to detect any changes in the liquid phase content and crystal form of the sample, and to record any changes in the color of the sample.

[0070] Table 4. High-temperature test results of samples from Examples 1-3

[0071] Table 5. High humidity test results of samples from Examples 1-3

[0072] Table 6 shows the results of the strong light irradiation test on the samples from Examples 1-3.

[0073] The results above show that the gingerone A crystals prepared by this invention can maintain high stability under high temperature, high humidity, and strong light irradiation conditions. In particular, the gingerone A content of the gingerone A single crystals only changed slightly after 10 days under high temperature, high humidity, and light irradiation conditions, which is significantly better than that of samples 2 and 3. This indicates that the gingerone A single crystals of this invention have excellent stability, and the single crystal form did not change and was very stable. In summary, the gingerone A crystals prepared by this invention have excellent stability and can be stored for a long time under relatively harsh conditions.

[0074] Example 5

[0075] Fifteen 6-8 week old male SD rats (188-200g) were randomly divided into four groups: Experimental Group 1 (shogaol A single crystal from Example 1), Experimental Group 2 (shogaol A crystal from Example 2), Experimental Group 3 (shogaol A crystal from Example 3), Control Group 1 (shogaol A oil), and a blank control group (equal volume of physiological saline), with three rats in each group. The rats were housed in transparent resin plastic cages (400mm×240mm×200mm) under fluorescent lighting, with 12 hours of light per day (07:00-19:00) and 12 hours of no light per day. The ambient temperature and relative humidity in the animal room were controlled within the ranges of 20-26℃ and 40-70%, respectively. All animals had free access to water during the experiment. The rats were fasted overnight before administration and fed again 4 hours after administration. Rats in the corresponding groups were administered 5 mg / kg of gingerone A single crystals from Example 1, gingerone A crystals from Example 2, gingerone A crystals from Example 3, gingerone A oil from Control Group 1, and an equal volume of physiological saline via gavage. Blood samples were collected before administration and at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 10 h, and 24 h after administration. After collection, blood samples were placed in labeled centrifuge tubes (pre-cooled with crushed ice) and rapidly centrifuged at 3500 rpm for 10 minutes at 4°C to separate plasma. The content of gingerone A in plasma was detected by high-performance liquid chromatography (HPLC). The concentration data of gingerone A in plasma were statistically analyzed using the WinNonlin metabolic data analysis software, and pharmacokinetic parameters were calculated using the non-compartmental model (NCA) method. The experimental results are shown in Table 7.

[0076] Table 7. Main pharmacokinetic parameters in rat plasma from experimental groups 1-3 and control group 1.

[0077] As can be seen from the above results, the gingerone A single crystal of the present invention can reach the peak concentration in a short time, is widely distributed in the rat body, and is not easily metabolized and cleared by the body, and can maintain an effective concentration in the body for a long time.

[0078] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A gingerone A crystal, including single crystals and polycrystalline forms, with E-trans-genone structural features and a melting point of 79-81℃. The melting point of crystals obtained by different methods varies slightly, generally between 79-80℃.

2. A single crystal of gingerone A, characterized in that, The unit cell parameters of the gingerone A single crystal are as follows:

3. The method for preparing gingerone A crystals according to claim 1, characterized in that, Includes the following steps: (1) Add gingerone A oil to an organic solvent of more than 2 times its weight, heat and dissolve to obtain a clear solution; (2) Cool the clear solution obtained in step (1) to room temperature, keep it at a constant temperature, and let the organic solvent slowly evaporate and precipitate single crystals, or keep it at a constant temperature between 0 and -20℃ to precipitate crystals. After filtration, filter cake is obtained. Wash the filter cake 2-3 times with recrystallization solvent and dry it to obtain the final product.

4. The preparation method according to claim 3, characterized in that, The organic solvents mentioned in steps (1) and (2) are one or a mixture of two or more of the following solvents: dichloromethane, methanol, 95% ethanol, 95% methanol, diethyl ether, methyl tert-butyl ether, ethyl acetate, n-heptane, petroleum ether at 60-90℃, n-hexane, cyclohexane, and toluene.

5. The preparation method according to claim 3, characterized in that, The gingerone A content in the gingerone A oily substance mentioned in step (1) is 95-98%.

6. The preparation method according to claim 3, characterized in that, The amount of organic solvent used in step (1) is 2 to 11 times the amount of gingerone A oil.

7. The preparation method according to claim 3, characterized in that, The temperature for dissolving the solution in step (1) is 30–70°C.

8. The preparation method according to claim 3, characterized in that, In step (2), the cooling rate is controlled at 0.1 to 2 °C / min.

9. The preparation method according to claim 3, characterized in that, The amount of recrystallization solvent used in step (2) is 2 to 10 times the amount of gingerone A oily substance; the drying includes forced air drying and vacuum drying, the drying temperature is 30 to 60°C, the vacuum degree of vacuum drying is ≤-0.085MPa, and the drying time is 5 to 24h.

10. The preparation method according to claim 4, characterized in that, In the organic solvent, the volume ratio of diethyl ether to methanol, ethanol, dichloromethane, ethyl acetate or methyl tert-butyl ether is from 1:1 to 10:

1.

11. The preparation method according to claim 4, characterized in that, In the organic solvent, the volume ratio of methyl tert-butyl ether to methanol, ethanol, dichloromethane or ethyl acetate is 5:1 to 10:

1.

12. The preparation method according to claim 4, characterized in that, In the organic solvent, the volume ratio of n-heptane, n-hexane, cyclohexane, or petroleum ether at 60-90℃ to dichloromethane, ethyl acetate, methyl tert-butyl ether, or toluene is 2:1 to 5:1.

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