Hydroxytyrosol Ectoin eutectic as well as preparation method and application thereof

Abstract

The invention discloses a hydroxytyrosol ectoine eutectic as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The molar ratio of hydroxytyrosol to Ectoine in the hydroxytyrosol Ectoine eutectic is 1: 1. The preparation method comprises the following steps: (1) weighing hydroxytyrosol and Ectoin according to a molar ratio, adding the hydroxytyrosol and Ectoin into a solvent, and carrying out heat preservation and stirring until the hydroxytyrosol and Ectoin are completely dissolved; (2) depressurizing the solution in the step (1) to remove the solvent to obtain a concentrated solution; and (3) carrying out low-temperature stirring crystallization on the concentrated solution, and drying to obtain the hydroxytyrosol Ectoine eutectic. The hydroxytyrosol Ectoin eutectic can be used for preparing anti-inflammatory drugs, vascular dilation drugs, antibacterial drugs or anti-cancer drugs, and can also be used for preparing moisturizing cosmetics, anti-aging cosmetics, anti-oxidation cosmetics, anti-wrinkle cosmetics and repair cosmetics. According to the hydroxytyrosol Ectoine eutectic prepared by the invention, the melting point of hydroxytyrosol can be increased from the molecular level, the stability of hydroxytyrosol is improved, and the application field of hydroxytyrosol is widened.

Description

Technical Field

[0001] The invention relates to a hydroxytyrosol-ectoine cocrystal and a preparation method and application thereof, belonging to the technical field of biomedicine. Background Art

[0002] Hydroxytyrosol is an active ingredient in olives and is considered one of the most powerful antioxidants. Hydroxytyrosol has multiple biological and pharmacological activities, including anti-inflammatory, vasodilatory and antibacterial effects. It is effective in the prevention and treatment of arteriosclerosis, hypertension, heart disease, cerebral hemorrhage, etc. It has potential medical effects and has been widely used in food, health products and cosmetics. However, hydroxytyrosol itself has a low melting point (52°C) and high hygroscopicity. It appears as a waxy solid or viscous oily liquid at ambient temperature and humidity. It is inconvenient to use and cannot be directly used in hard capsules or tablets. The raw material itself is extremely unstable to light, heat, moisture, oxygen, etc., and is easily oxidized and discolored during use.

[0003] At present, the encapsulation method is generally used to prepare solid powder to protect the stability and activity of hydroxytyrosol. The hydroxytyrosol encapsulation powder has a loading of 20-30%. However, due to the amphiphilic nature of hydroxytyrosol, the encapsulation effect is poor, and the unencapsulated hydroxytyrosol still has problems such as instability and easy discoloration. In addition, since the encapsulation powder contains a large amount of starch excipients, its hygroscopicity is further increased.

[0004] Therefore, it is very important to provide a suitable method to increase the melting point of hydroxytyrosol and improve its stability. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention aims to provide a hydroxytyrosol-ectoine cocrystal and a preparation method and application thereof. By preparing the hydroxytyrosol-ectoine cocrystal, the melting point of hydroxytyrosol can be increased at the molecular level, and its stability can be improved, thereby broadening its application field.

[0006] In order to solve the above technical problems, the technical solutions provided by the present invention are as follows: A hydroxytyrosol-ectoine co-crystal, wherein the molar ratio of hydroxytyrosol to ectoine in the hydroxytyrosol-ectoine co-crystal is 1:1.

[0007] Furthermore, the X-ray powder diffraction of the hydroxytyrosol ectoine cocrystal expressed in 2θ angles has characteristic peaks at least at diffraction angles of 8.6±0.2, 11.3±0.2, 13.6±0.2, 14.6±0.2, 15.0±0.2, 17.9±0.2, 19.9±0.2, 20.9±0.2, 21.5±0.2, 22.4±0.2, 22.8±0.2, 23.7±0.2, 24.7±0.2, 26.2±0.2, and 29.4±0.2°.

[0008] Further, the hydroxytyrosol ectoine cocrystal is prepared by Figure 1 The X-ray powder diffraction pattern is shown.

[0009] Furthermore, the melting point of the hydroxytyrosol-ectoine eutectic is 133-137°C.

[0010] A method for preparing a hydroxytyrosol-ectoine cocrystal comprises the following steps: (1) Weigh hydroxytyrosol and ectoine in a molar ratio and add them to the solvent, stirring at room temperature until they are completely dissolved; (2) reducing the pressure of the solution in step (1) to remove the solvent to obtain a concentrated solution; (3) stirring the concentrated solution at low temperature for crystallization, and drying it to obtain hydroxytyrosol-ectoine cocrystals.

[0011] Furthermore, in step (1), the molar ratio of hydroxytyrosol to ectoine is 1:(0.5-1.5), preferably 1:1; the solvent is one or more of water, methanol or ethanol, preferably methanol; and the temperature of heat preservation and stirring is 35-45°C.

[0012] Furthermore, in step (2), the depressurization operation for removing the solvent is: concentrating the solution by vacuum evaporation using a rotary evaporator; the vacuum degree of the vacuum evaporation concentration is less than -0.08 MPa, the concentration temperature is 35-45°C, and the concentration is terminated when the concentrated solution becomes viscous and the solvent content is less than 20% (m / m).

[0013] Furthermore, the vacuum degree of the vacuum evaporation concentration is less than -0.09Mpa, the concentration temperature is 35-40°C, and the concentration is terminated when the concentrated liquid becomes viscous and the solvent content is less than 10% (m / m).

[0014] Furthermore, the solvent content is measured using a rapid moisture analyzer, with the measurement parameters set at 105° C. and 10 min.

[0015] Further, in step (3), the concentrated solution is taken out from the concentration bottle, placed in a beaker pre-cooled on ice, placed in an ice bath, stirred at 100-200 rpm / min for 30-60 min for crystallization, the crystals are collected and ground into fine powder in a clean mortar, and dried to constant weight under vacuum degree <-0.08 MPa and temperature of 40-45°C to obtain hydroxytyrosol ectoine cocrystals.

[0016] An application of a hydroxytyrosol-ectoine cocrystal, which can be used to prepare anti-inflammatory drugs, vasodilator drugs, antibacterial drugs or anticancer drugs. The dosage form of the drug is preferably powder, tablet, granule, capsule, solution, emulsion, suspension, injection, spray, aerosol or powder spray; the drug also preferably contains other excipients and / or active ingredients, and the present invention has no special requirements on the type of the excipients, as long as they do not affect the activity of the hydroxytyrosol-ectoine cocrystal; the drug also preferably contains a pharmaceutically acceptable carrier, and the carrier includes a diluent, a buffer, a suspension, an emulsion, a granule, an encapsulation agent, an excipient, a filler, an adhesive, a spray, a transdermal absorbent, a wetting agent, a disintegrant, an absorption promoter, a surfactant, a colorant, a flavoring agent or an adsorption carrier.

[0017] The application of the hydroxytyrosol and ectoine co-crystal can also be used to prepare moisturizing cosmetics, anti-aging cosmetics, antioxidant cosmetics, anti-wrinkle cosmetics, and repair cosmetics. The dosage form of the cosmetic is preferably a cream, a facial mask, an emulsion, a toner, a shower gel, a facial cleanser, or a hand sanitizer / hand washing gel; the cosmetic also includes auxiliary materials acceptable to cosmetics, and the auxiliary materials include a moisturizer, an emulsifier, a thickener, a skin conditioner, a pH adjuster, a preservative, and water.

[0018] Pharmaceutical cocrystal refers to the active pharmaceutical ingredient (API) and other physiologically acceptable ligands that are connected in a fixed stoichiometric ratio through hydrogen bonds and other non-covalent bonds and combined in the same crystal lattice. It can change the physicochemical properties of the drug without changing the API structure, thereby affecting its physiological activity. Changing the solid form of the API through cocrystal technology is an important means to improve the physicochemical properties of the drug, such as solubility, permeability, hygroscopicity, bioavailability, melting point and stability. Therefore, the present invention prepares the hydroxytyrosol ectoine cocrystal through the cocrystal technology, and increases the melting point of hydroxytyrosol, weakens its hygroscopicity and improves its stability without changing the structure of hydroxytyrosol.

[0019] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention prepares hydroxytyrosol-ectoine eutectic by eutectic technology, thereby increasing the melting point of hydroxytyrosol, reducing its hygroscopicity, and improving its stability without changing the structure of hydroxytyrosol, thereby solving the problems of hydroxytyrosol with low melting point, high hygroscopicity, and extreme instability to heat, moisture, oxygen, etc.

[0020] (2) Compared with hydroxytyrosol itself, the melting point of the hydroxytyrosol-ectoine co-crystal of the present invention is significantly increased. Compared with hydroxytyrosol itself and its embedded powder, the hydroxytyrosol-ectoine co-crystal of the present invention has reduced hygroscopicity and significantly improved chemical stability.

[0021] (3) The preparation method of the hydroxytyrosol-ectoine cocrystal provided by the present invention has simple steps, high efficiency, low cost, and is suitable for large-scale production. It can expand the application field of hydroxytyrosol and increase the economic benefits of the industry. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 1 is an X-ray powder diffraction pattern of the hydroxytyrosol ectoine co-crystal of the embodiment of the present invention. DETAILED DESCRIPTION

[0023] The principles and features of the present invention are described below. The examples are only used to explain the present invention and are not used to limit the scope of the present invention. At the same time, the experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents, materials, and instruments are all prior art and can be obtained from commercial channels unless otherwise specified.

[0024] The X-ray powder diffraction pattern in the embodiment of the present invention is obtained by using a PANalytical Empyream model X-ray powder diffractometer, and the main test parameters are 40 kV, 40 mA; Hydroxytyrosol and Ectoin are both from Shandong Freda Biological Company, with a purity of ≥98%; the hydroxytyrosol embedding powder contains 25% hydroxytyrosol and 75% maltodextrin.

[0025] Example 1 (1) Weigh 100 mmol of hydroxytyrosol and 100 mmol of ectoine respectively, add them into a beaker containing 350 mL of anhydrous methanol, and stir in a 45°C water bath until completely dissolved; (2) The solution in step (1) was transferred to a rotary evaporator for concentration at a vacuum degree of -0.095 MPa and a concentration temperature of 45°C. The concentration was terminated when the concentrated solution became viscous and the methanol content was 7.3% (m / m), thereby obtaining a concentrated solution. (3) The concentrated solution was transferred to a beaker precooled on ice, placed in an ice bath, and stirred at 100 rpm / min for 30 min to allow crystallization. The crystals were collected and ground, and then dried at a vacuum degree of -0.09 MPa and a temperature of 40°C to constant weight to obtain hydroxytyrosol ectoine cocrystals.

[0026] Example 2 (1) Weigh 30 mmol of hydroxytyrosol and 30 mmol of ectoine respectively, add them into a beaker containing 400 mL of anhydrous ethanol, and stir in a 45°C water bath until completely dissolved; (2) The solution in step (1) was transferred to a rotary evaporator for concentration at a vacuum degree of -0.095 MPa and a concentration temperature of 45°C. The concentration was terminated when the concentrated solution became viscous and the ethanol content was 9.8% (m / m), thereby obtaining a concentrated solution. (3) The concentrated solution was transferred to a beaker precooled on ice, placed in an ice bath, and stirred at 150 rpm / min for 40 min for crystallization. The crystals were collected and ground, and then dried at a vacuum degree of -0.09 MPa and a temperature of 40°C to constant weight to obtain hydroxytyrosol ectoine cocrystals.

[0027] Example 3 (1) Weigh 50 mmol of hydroxytyrosol and 50 mmol of ectoine respectively, add them into a beaker containing 400 mL of a mixed solvent (anhydrous methanol and anhydrous ethanol mixed in a volume ratio of 1:1), and stir in a 45°C water bath until completely dissolved; (2) The solution in step (1) was transferred to a rotary evaporator for concentration at a vacuum degree of -0.095 MPa and a concentration temperature of 45°C. The concentration was terminated when the concentrated solution became viscous and the solvent content was 8.5% (m / m), thereby obtaining a concentrated solution. (3) The concentrated solution was transferred to a beaker precooled on ice, placed in an ice bath, and stirred at 130 rpm / min for 30 min to crystallize. The crystals were collected and ground, and then dried at a vacuum degree of -0.09 MPa and a temperature of 40°C to constant weight to obtain hydroxytyrosol ectoine cocrystals.

[0028] Test Example 1 The hydroxytyrosol ectoine cocrystal solid powder obtained in Examples 1-3 was tested using a powder diffractometer PANalytical Empyream at 40 kV and 40 mA in a scanning range of 0° to 60° in the 2θ interval.

[0029] After testing, it can be seen that the solid powder diffraction results of the hydroxytyrosol ectoin cocrystal obtained in Examples 1-3 are consistent, and the X-ray powder diffraction pattern is as follows Figure 1As shown, the X-ray powder diffraction patterns expressed in 2θ angles all have characteristic peaks at least at diffraction angles of 8.6±0.2, 11.3±0.2, 13.6±0.2, 14.6±0.2, 15.0±0.2, 17.9±0.2, 19.9±0.2, 20.9±0.2, 21.5±0.2, 22.4±0.2, 22.8±0.2, 23.7±0.2, 24.7±0.2, 26.2±0.2, and 29.4±0.2°.

[0030] By analyzing the spectra and data, it can be seen that the characteristic peaks of the hydroxytyrosol ectoine co-crystals obtained in Examples 1-3 are significantly different from the reported characteristic peaks of ectoine (11.53±0.2, 12.12±0.2, 13.45±0.2, 15.52±0.2, 16.87±0.2, 17.92±0.2, 19.49±0.2, 20.40±0.2, 21.39±0.2, 23.12±0.2, 24.43±0.2, 25.04±0.2, 25.78±0.2, 26.55±0.2, 27.03±0.2, 28.58±0.2, 30.94±0.2°).

[0031] Every kind of crystal corresponds to different powder diffraction distributions, and the distribution and intensity of diffraction lines have characteristic regularity, which plays a decisive role in whether there is a new crystalline phase to produce the stage. When forming a new phase, the X-ray powder diffraction pattern will be different from the bulk drug and the ligand. New phases are produced by PXRD spectrum identification, which can be analyzed and identified. From the results of the sample X-ray powder diffraction patterns of embodiment 1-3, it can be judged that hydroxytyrosol ectoin eutectic substances have been formed.

[0032] Test Example 2 The hydroxytyrosol and ectoine contents of the hydroxytyrosol-ectoine cocrystal solid powder obtained in Example 1 were determined by HPLC, and the molar ratio was calculated.

[0033] Table 1. Composition analysis of hydroxytyrosol and ectoine cocrystals Hydroxytyrosol (molar number): Ectoin (molar number) = (hydroxytyrosol content / 154.16): (ectoin content / 142.16).

[0034] Substituting the data in Table 1 into the calculation formula, it can be seen that the molar ratio of hydroxytyrosol and ectoine in the hydroxytyrosol-ectoine cocrystal is 1:1.

[0035] Test Example 3 Referring to the fourth part, general chapter 0612, melting point determination method, method one of the 2020 edition of the "Chinese Pharmacopoeia", the melting point of the hydroxytyrosol-ectoine co-crystal was determined to be 133-137°C, while the reported melting points of hydroxytyrosol and ectoine were 52°C and 280°C, respectively. The melting point of the hydroxytyrosol-ectoine co-crystal was different from those of both, confirming the formation of a new phase in the co-crystal.

[0036] Test Example 4 With reference to Part IV, General Chapter 9103, Guiding Principles for Drug Hygroscopicity Tests of the 2015 Edition of the Chinese Pharmacopoeia, the hygroscopicity of hydroxytyrosol and the hydroxytyrosol-ectoine co-crystal obtained in the examples was determined and compared.

[0037] Table 2. Hygroscopicity of Hydroxytyrosol-Ectoin Cocrystals It can be seen from the results in Table 2 that the weight gain percentage of hydroxytyrosol is 15.2%, which is extremely hygroscopic; the weight gain percentage of hydroxytyrosol-ectoine co-crystal prepared by the present invention is 0.3%, which is slightly hygroscopic, indicating that the present invention adopts the co-crystal technology to prepare hydroxytyrosol-ectoine co-crystal, which can significantly improve the hygroscopicity of hydroxytyrosol, which is beneficial to the chemical stability of hydroxytyrosol under high humidity conditions.

[0038] Test Example 5 Take 30 portions of pure hydroxytyrosol, hydroxytyrosol embedded powder and hydroxytyrosol ectoine cocrystal, each about 100 mg, and package them separately in double-layer polyethylene ziplock bags and put them into an accelerated stability box (acceleration conditions are 50℃ / 80% RH). Sampling was carried out at 0 days, 15 days, 30 days, 60 days and 120 days respectively, and the residual hydroxytyrosol content in the samples was analyzed by HPLC. The hydroxytyrosol content in the initial samples was set as 100%, and the ratio of the hydroxytyrosol HPLC content measured at different times and the hydroxytyrosol HPLC content measured initially was recorded as the residual hydroxytyrosol content at the corresponding time. 。

[0039] Table 3. Stable experimental contents of hydroxytyrosol, hydroxytyrosol embedded powder and hydroxytyrosol ectoin cocrystal (50℃ / 80%RH) As shown in Table 3, the initial hydroxytyrosol content in the sample is all set to 100%. After 120 days, the content of hydroxytyrosol in the hydroxytyrosol pure product is 46.5% of the initial content, and the content of hydroxytyrosol in the hydroxytyrosol embedded powder is 27.2% of the initial content, while in the hydroxytyrosol Ecdoin eutectic provided by the present invention, the content of hydroxytyrosol is 99.5% of the initial content, still remaining above 99%. It can be seen that, compared with the hydroxytyrosol pure product and the hydroxytyrosol embedded powder, the hydroxytyrosol Ecdoin eutectic of the present invention has better chemical stability.

Claims

1. A hydroxytyrosol ectoin eutectic, characterized in that: The molar ratio of hydroxytyrosol to ectoine in the hydroxytyrosol-ectoine co-crystal is 1:

1.

2. The hydroxytyrosol ectoin co-crystal according to claim 1, characterized in that: The X-ray powder diffraction of the hydroxytyrosol ectoine cocrystal expressed in 2θ angles has characteristic peaks at least at diffraction angles of 8.6±0.2, 11.3±0.2, 13.6±0.2, 14.6±0.2, 15.0±0.2, 17.9±0.2, 19.9±0.2, 20.9±0.2, 21.5±0.2, 22.4±0.2, 22.8±0.2, 23.7±0.2, 24.7±0.2, 26.2±0.2, and 29.4±0.2°.

3. The hydroxytyrosol ectoin co-crystal according to claim 1, characterized in that: The hydroxytyrosol ectoine co-crystal is characterized by an X-ray powder diffraction pattern as shown in FIG1 .

4. The hydroxytyrosol ectoin co-crystal according to claim 1, characterized in that: The melting point of the hydroxytyrosol ectoine eutectic is 133-137°C.

5. A method for preparing the hydroxytyrosol ectoine co-crystal according to claim 1, characterized in that: The following steps are involved: (1) Weigh hydroxytyrosol and ectoine in a molar ratio and add them to the solvent, stirring at room temperature until they are completely dissolved; (2) reducing the pressure of the solution in step (1) to remove the solvent to obtain a concentrated solution; (3) stirring the concentrated solution at low temperature for crystallization, and drying it to obtain hydroxytyrosol-ectoine cocrystals.

6. The method for preparing the hydroxytyrosol ectoin cocrystal according to claim 5, wherein: In step (1), the molar ratio of hydroxytyrosol to ectoine is 1:(0.5-1.5), preferably 1:1; the solvent is one or more of water, methanol or ethanol, preferably methanol; and the temperature of heat preservation and stirring is 35-45°C.

7. The method for preparing the hydroxytyrosol ectoin co-crystal according to claim 5, wherein: In step (2), the depressurization operation for removing the solvent is as follows: the solution is concentrated by vacuum evaporation using a rotary evaporator; the vacuum degree of the vacuum evaporation concentration is less than -0.08 MPa, the concentration temperature is 35-45°C, and the concentration is terminated when the concentrated solution becomes viscous and the solvent content is less than 20% (m / m).

8. The method for preparing the hydroxytyrosol ectoin cocrystal according to claim 7, wherein: The vacuum degree of the vacuum evaporation concentration is less than -0.09Mpa, the concentration temperature is 35-40°C, and the concentration is terminated when the concentrated liquid becomes viscous and the solvent content is less than 10% (m / m).

9. The method for preparing the hydroxytyrosol ectoin cocrystal according to claim 5, wherein: Step (3), taking out the concentrated solution from the concentration bottle, placing it into a beaker pre-cooled on ice, placing it in an ice bath, stirring at 100-200 rpm / min for 30-60 min to crystallize, collecting the crystals and grinding them, and drying them to constant weight under the conditions of vacuum degree <-0.08 MPa and temperature of 40-45° C. to obtain hydroxytyrosol ectoine cocrystals.

10. An application of the hydroxytyrosol ectoine co-crystal according to claim 1, characterized in that: The hydroxytyrosol ectoine cocrystal can be used to prepare anti-inflammatory drugs, vasodilator drugs, antibacterial drugs or anticancer drugs, and can also be used to prepare moisturizing cosmetics, anti-aging cosmetics, antioxidant cosmetics, anti-wrinkle cosmetics, and repair cosmetics.

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