A hydroxytyrosol-ecdoine cocrystal and its preparation method and application

Abstract

The present invention discloses a hydroxytyrosol-ectoine cocrystal and its preparation method and application, belonging to the field of biomedicine technology. The molar ratio of hydroxytyrosol to ectoine in the hydroxytyrosol-ectoine cocrystal is 1:1. The preparation method comprises the following steps: (1) weighing hydroxytyrosol and ectoine in molar ratio and adding them to a solvent, keeping warm and stirring until 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 a low temperature for crystallization and drying to obtain a hydroxytyrosol-ectoine cocrystal. 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. The preparation of the hydroxytyrosol-ectoine cocrystal by the present invention can increase the melting point of hydroxytyrosol at the molecular level, improve its stability, and broaden its application field.

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, an active ingredient in olives, is considered one of the most powerful antioxidants. Hydroxytyrosol exhibits multiple biological and pharmacological activities, including anti-inflammatory, vasodilatory, and antibacterial properties. It is effective in the prevention and treatment of arteriosclerosis, hypertension, heart disease, cerebral hemorrhage, and other conditions, showing potential medical benefits. It has also been widely used in food, health supplements, and cosmetics. However, hydroxytyrosol itself has a low melting point (52°C) and is highly hygroscopic. At ambient temperature and humidity, it forms a waxy solid or a viscous, oily liquid, making it inconvenient to handle and unable to be used directly in hard capsules or tablets. Furthermore, the raw material is extremely unstable to light, heat, moisture, and oxygen, and is prone to oxidation and discoloration during use.

[0003] Currently, the encapsulation method is commonly used to prepare solid powders to protect the stability and activity of hydroxytyrosol. The resulting hydroxytyrosol-encapsulated powder has a loading of 20-30%. However, due to the amphiphilic nature of hydroxytyrosol, the encapsulation effect is poor, and unencapsulated hydroxytyrosol still has problems such as instability and easy discoloration. In addition, since the encapsulated 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 its preparation method and application. 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 present invention provides the following technical solutions:

[0007] A hydroxytyrosol-ectoine co-crystal, wherein the molar ratio of hydroxytyrosol to ectoine is 1:1.

[0008] 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°.

[0009] Further, the hydroxytyrosol ectoine cocrystal is prepared by Figure 1 The X-ray powder diffraction pattern is shown.

[0010] Furthermore, the melting point of the hydroxytyrosol-ecdoine eutectic is 133-137°C.

[0011] A method for preparing a hydroxytyrosol-ectoine cocrystal comprises the following steps:

[0012] (1) Weigh hydroxytyrosol and ectoine in molar ratio and add them to the solvent, stirring at room temperature until they are completely dissolved;

[0013] (2) reducing the pressure of the solution in step (1) to remove the solvent to obtain a concentrated solution;

[0014] (3) The concentrated solution is stirred and crystallized at low temperature and dried to obtain hydroxytyrosol-ectoine cocrystal.

[0015] 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.

[0016] Furthermore, 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).

[0017] Furthermore, the vacuum degree of the vacuum evaporation concentration is less than -0.09 MPa, 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).

[0018] Furthermore, the solvent content is measured using a rapid moisture analyzer, with the measurement parameters set at 105° C. and 10 min.

[0019] Furthermore, in step (3), the concentrate is taken out from the concentrate bottle, placed in a beaker pre-cooled on ice, placed in an ice bath, and 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. The powder is dried to constant weight under vacuum degree <-0.08 MPa and temperature of 40-45°C to obtain hydroxytyrosol ectoine cocrystals.

[0020] A use of a hydroxytyrosol-ectoine co-crystal, which can be used to prepare anti-inflammatory drugs, vasodilator drugs, antibacterial drugs, or anticancer drugs. The drug is preferably in the form of a powder, tablet, granule, capsule, solution, emulsion, suspension, injection, spray, aerosol, or powder spray; the drug also preferably contains other excipients and / or active ingredients. The present invention has no special requirements for the type of the excipient, as long as it does not affect the activity of the hydroxytyrosol-ectoine co-crystal; the drug also preferably contains a pharmaceutically acceptable carrier, including a diluent, buffer, suspension, emulsion, granule, encapsulating agent, excipient, filler, adhesive, spray, transdermal absorbent, wetting agent, disintegrant, absorption enhancer, surfactant, colorant, flavoring agent, or adsorption carrier.

[0021] The hydroxytyrosol and ectoine co-crystals can also be used to prepare moisturizing cosmetics, anti-aging cosmetics, antioxidant cosmetics, anti-wrinkle cosmetics, and repair cosmetics. The cosmetics are preferably in the form of a cream, facial mask, lotion, toner, shower gel, facial cleanser, or hand soap / hand gel; the cosmetics also include cosmetically acceptable excipients, including a moisturizer, emulsifier, thickener, skin conditioner, pH adjuster, preservative, and water.

[0022] A pharmaceutical cocrystal is a combination of an active pharmaceutical ingredient (API) and another physiologically acceptable ligand bound in a fixed stoichiometric ratio via hydrogen bonds and other non-covalent bonds within a single crystal lattice. This cocrystal can alter the drug's physicochemical properties, thereby affecting its physiological activity, without altering the API's structure. Modifying the solid form of an API through cocrystal technology is an important means of improving drug properties such as solubility, permeability, hygroscopicity, bioavailability, melting point, and stability. Therefore, the present invention utilizes cocrystal technology to prepare hydroxytyrosol-ectoine cocrystals, raising the melting point, reducing hygroscopicity, and enhancing stability of hydroxytyrosol without altering its structure.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] (1) The present invention prepares hydroxytyrosol-ecdoine eutectic by eutectic technology, which increases the melting point of hydroxytyrosol, reduces its hygroscopicity, and improves 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.

[0025] (2) Compared to hydroxytyrosol itself, the melting point of the hydroxytyrosol-ectoine co-crystal of the present invention is significantly increased. Compared to hydroxytyrosol itself and its embedded powder, the hydroxytyrosol-ectoine co-crystal of the present invention has reduced hygroscopicity and significantly improved chemical stability.

[0026] (3) The preparation method of the hydroxytyrosol-ecdoine 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

[0027] Figure 1 1 is an X-ray powder diffraction pattern of the hydroxytyrosol ectoine cocrystal according to an embodiment of the present invention. DETAILED DESCRIPTION

[0028] The principles and features of the present invention are described below. The examples provided are intended only to illustrate the present invention and are not intended to limit its scope. The experimental methods described in the following examples are conventional methods unless otherwise specified. The reagents, materials, and instruments described are all prior art and commercially available unless otherwise specified.

[0029] The X-ray powder diffraction patterns in the embodiments of the present invention were obtained using a PANalytical Empyream model X-ray powder diffractometer, with the main test parameters being 40 kV and 40 mA.

[0030] Hydroxytyrosol and ectoine were both sourced from Shandong Freda Biological Company, with a purity of ≥98%. The hydroxytyrosol embedding powder contained 25% hydroxytyrosol and 75% maltodextrin.

[0031] Example 1

[0032] (1) Weigh 100 mmol of hydroxytyrosol and 100 mmol of ectoine separately and add them into a beaker containing 350 mL of anhydrous methanol. Stir in a 45°C water bath until completely dissolved.

[0033] (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 completed when the concentrated solution became viscous and the methanol content was 7.3% (m / m). A concentrated solution was obtained.

[0034] (3) The concentrated solution was transferred to a beaker pre-cooled on ice, placed in an ice bath, and stirred at 100 rpm / min for 30 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.

[0035] Example 2

[0036] (1) Weigh 30 mmol of hydroxytyrosol and 30 mmol of ectoine separately and add them into a beaker containing 400 mL of anhydrous ethanol. Stir in a 45°C water bath until completely dissolved.

[0037] (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 completed when the concentrated solution became viscous and the ethanol content was 9.8% (m / m). A concentrated solution was obtained.

[0038] (3) The concentrated solution was transferred to a beaker pre-cooled 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.

[0039] Example 3

[0040] (1) Weigh 50 mmol of hydroxytyrosol and 50 mmol of ectoine separately and add them into a beaker containing 400 mL of 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;

[0041] (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 completed when the concentrated solution became viscous and the solvent content was 8.5% (m / m). A concentrated solution was obtained.

[0042] (3) The concentrated solution was transferred to a beaker pre-cooled on ice, placed in an ice bath, and stirred at 130 rpm / min for 30 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.

[0043] Test Example 1

[0044] The hydroxytyrosol-ectoine cocrystal solid powders obtained in Examples 1-3 were 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.

[0045] After testing, it can be seen that the solid powder diffraction results of the hydroxytyrosol ectoine 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 degrees 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°.

[0046] Analysis of the spectra and data shows that the characteristic peaks of the hydroxytyrosol-ecdoine co-crystals obtained in Examples 1-3 are significantly different from the reported characteristic peaks of ecdoine (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°).

[0047] Every kind of crystal is corresponding to different powder diffraction distributions, and the distribution and intensity of diffraction line have characteristic regularity, plays a decisive role in whether new crystalline phase produces stage.When forming new phase, X-ray powder diffraction pattern will be different from bulk drug and part.Produce new phase by PXRD spectrum identification, it can be analyzed and differentiated.Can judge from embodiment 1-3 sample X-ray powder diffraction pattern result, formed hydroxytyrosol ectoine eutectic thing.

[0048] Test Example 2

[0049] 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.

[0050] Table 1. Composition analysis of hydroxytyrosol and ectoine cocrystals

[0051]

[0052] Hydroxytyrosol (molar number): Ectoin (molar number) = (Hydroxytyrosol content / 154.16): (Ectoin content / 142.16).

[0053] 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.

[0054] Test Example 3

[0055] Referring to the 2020 edition of the "Chinese Pharmacopoeia", Part 4, General Chapter 0612, Melting Point Determination Method, Method 1, the melting point of the hydroxytyrosol-ectoine cocrystal was determined to be 133-137°C, while the reported melting point of hydroxytyrosol was 52°C and the melting point of ectoine was 280°C. The melting point of the hydroxytyrosol-ectoine cocrystal was different from the melting points of both, confirming the formation of a new phase in the cocrystal.

[0056] Test Example 4

[0057] With reference to the Guiding Principles for Hygroscopicity Test of Drugs, Part IV, General Chapter 9103, 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.

[0058] Table 2. Hygroscopicity of Hydroxytyrosol-Ectoin Cocrystals

[0059]

[0060] As can be seen from the results in Table 2, the weight gain percentage of hydroxytyrosol is 15.2%, which is extremely hygroscopic; the weight gain percentage of the hydroxytyrosol-ectoine co-crystal prepared by the present invention is 0.3%, which is slightly hygroscopic, indicating that the hydroxytyrosol-ectoine co-crystal prepared by the present invention using the eutectic technology can significantly improve the hygroscopicity of hydroxytyrosol, which is beneficial to the chemical stability of hydroxytyrosol under high humidity conditions.

[0061] Test Example 5

[0062] Thirty portions (approximately 100 mg each) of pure hydroxytyrosol, hydroxytyrosol-encapsulated powder, and hydroxytyrosol-ectoine co-crystal were individually packaged in double-layer polyethylene ziplock bags and placed in an accelerated stability chamber (accelerated conditions: 50°C / 80% RH). Samples were collected at 0, 15, 30, 60, and 120 days, and the residual hydroxytyrosol content in the samples was analyzed by HPLC. The initial hydroxytyrosol content in each sample was assumed to be 100%. The ratio of the hydroxytyrosol HPLC content measured at subsequent times to the initial HPLC content was recorded as the residual hydroxytyrosol content at the corresponding time.

[0063] Table 3. Stability test contents of hydroxytyrosol, hydroxytyrosol embedded powder, and hydroxytyrosol-ecdotine cocrystal (50°C / 80%RH)

[0064]

[0065] As shown in Table 3, initial hydroxytyrosol content in sample is all determined to be 100%. After 120 days, the content of hydroxytyrosol in hydroxytyrosol pure product is 46.5% of initial content, and in hydroxytyrosol embedding powder, the content of hydroxytyrosol is 27.2% of initial content, while in hydroxytyrosol Ectoin eutectic provided by the present invention, the content of hydroxytyrosol is 99.5% of initial content, still remains on more than 99%. This shows that, compared with hydroxytyrosol pure product and hydroxytyrosol embedding powder, hydroxytyrosol Ectoin eutectic of the present invention has better chemical stability.

Claims

1. A hydroxytyrosol ectoine eutectic, characterized in that: The molar ratio of hydroxytyrosol and ectoine in the hydroxytyrosol-ectoine co-crystal is 1:1; the X-ray powder diffraction of the hydroxytyrosol-ectoine co-crystal 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°.

2. The hydroxytyrosol ectoine eutectic according to claim 1, wherein: The hydroxytyrosol-ectoine co-crystal is characterized by the X-ray powder diffraction pattern shown in FIG1 .

3. The hydroxytyrosol ectoine eutectic according to claim 1, wherein: The melting point of the hydroxytyrosol ectoine eutectic is 133-137°C.

4. A method for preparing the hydroxytyrosol ectoine cocrystal according to claim 1, characterized in that: The following steps are involved: (1) Weigh hydroxytyrosol and ectoine in 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) The concentrated solution is stirred and crystallized at low temperature and dried to obtain hydroxytyrosol-ectoine cocrystal.

5. the preparation method of hydroxytyrosol ectoine eutectic according to claim 4, is characterized in that: In step (1), the molar ratio of hydroxytyrosol to ectoine is 1:(0.5-1.5); the solvent is one or more of water, methanol or ethanol; and the temperature for heat preservation and stirring is 35-45°C.

6. the preparation method of hydroxytyrosol ectoine eutectic according to claim 4, is characterized in that: In step (2), the decompression 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).

7. The preparation method of the hydroxytyrosol ectoine eutectic according to claim 4, wherein: The vacuum evaporation concentration has a vacuum degree of less than -0.09 MPa and a concentration temperature of 35-40° C. The concentration is terminated when the concentrated liquid becomes viscous and the solvent content is less than 10% (m / m).

8. the preparation method of hydroxytyrosol ectoine eutectic according to claim 4, is characterized in that: 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 to crystallize, the crystals are collected and ground, and then dried to constant weight under the conditions of vacuum degree <-0.08 MPa and temperature of 40-45°C to obtain hydroxytyrosol ectoine cocrystals.

9. An application of the hydroxytyrosol ectoine cocrystal 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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