A ternary eutectic solvent, a preparation method and application thereof
By preparing a ternary eutectic solvent and using hydrogen bond donors and acceptors to form adenosine, the problem of low solubility of adenosine in water was solved, achieving high efficiency in dissolving and bioavailability of adenosine, simplifying the production process and reducing costs.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SHENZHEN SHINESKY BIOLOGICAL TECH CO LTD
- Filing Date
- 2025-05-21
- Publication Date
- 2026-06-23
AI Technical Summary
Adenosine has low solubility in water, which slows down the dissolution and absorption of the drug in the body, thus affecting its efficacy.
A ternary eutectic solvent system is prepared by heating and stirring, consisting of hydrogen bond donors (such as malic acid, tartaric acid, citric acid, and salicylic acid) and hydrogen bond acceptors (such as L-carnitine and betaine) with adenosine, which significantly improves the solubility and bioavailability of adenosine.
It significantly increases the solubility of adenosine by approximately 100 times, promotes rapid drug delivery to the target site, prolongs the duration of drug action, reduces production costs, and facilitates large-scale production.
Smart Images
Figure CN120661440B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of supramolecular technology, and in particular to a ternary eutectic solvent, its preparation method, and its application. Background Technology
[0002] Adenosine is an endogenous nucleoside found throughout human cells, possessing a wide range of physiological functions. For example, adenosine regulates the cardiovascular system, exhibiting effects such as vasodilation, blood pressure reduction, and antiarrhythmic activity; in the nervous system, it participates in sleep regulation and neuroprotection; and it also plays a crucial regulatory role in the immune system. However, adenosine has low solubility in water, which significantly limits its application in pharmaceutical formulations and its bioavailability. The low solubility of adenosine results in slow and incomplete dissolution and absorption of the drug in the body, making it difficult for the drug to effectively reach its target site, thus affecting its therapeutic efficacy.
[0003] Currently, common methods to improve drug solubility and bioavailability include salt formation, the use of surfactants, cyclodextrin inclusion complexation, and nanoparticle technology. However, these methods have many limitations. Salt formation may alter the chemical properties and stability of the drug, and not all drugs can have their solubility effectively improved through salt formation; the use of surfactants may cause toxic side effects and has limited solubilizing effects; cyclodextrin inclusion complexation requires strict control over the type and ratio of cyclodextrins, is costly, and has unstable inclusion rates; while nanoparticle technology can significantly improve drug solubility and bioavailability, its preparation process is complex, requires specialized equipment and technology, and is costly, hindering large-scale production and clinical application.
[0004] Therefore, existing technologies still need to be improved and developed. Summary of the Invention
[0005] In view of the shortcomings of the prior art, the present invention provides a ternary eutectic solvent, its preparation method and application, thereby solving the problem of low solubility of adenosine in water.
[0006] The technical solution adopted by the present invention to solve the above-mentioned technical problems is as follows:
[0007] In a first aspect, the present invention provides a ternary eutectic solvent, the ternary eutectic solvent being prepared from components A, B and C, wherein component A is a hydrogen bond donor, component B is a hydrogen bond acceptor and component C is adenosine.
[0008] Preferably, component A is selected from one or more of malic acid, tartaric acid, citric acid, and salicylic acid.
[0009] Preferably, component B is selected from one or both of L-carnitine and betaine.
[0010] Preferably, in the ternary eutectic solvent, the molar ratio of component A, component B and component C is (5-15):(5-15):1.
[0011] Preferably, in the ternary eutectic solvent, the molar ratio of component A, component B and component C is (8-12):(8-12):1.
[0012] A second aspect of the present invention provides a method for preparing the above-mentioned ternary eutectic solvent, the method comprising the following steps:
[0013] Component A and component B are mixed and subjected to a first heating and stirring process to obtain a mixed solution;
[0014] Component C is added to the above mixed solution, and a second heating and stirring treatment is performed to obtain the ternary eutectic solvent.
[0015] Preferably, the temperature of the first heating and stirring treatment is 50-80°C, and the time is 1-3 hours.
[0016] Preferably, the temperature of the second heating and stirring treatment is 35-90°C, and the time is 3-6 hours.
[0017] In a third aspect, the present invention provides the application of the above-described ternary eutectic solvent or the ternary eutectic solvent prepared by the above-described preparation method in the preparation of a drug.
[0018] Beneficial effects:
[0019] This invention discloses a ternary eutectic solvent, its preparation method, and its application. The ternary eutectic solvent provided by this invention has the following advantages: (1) Significantly improves adenosine solubility: The ternary eutectic solvent provided by this invention can increase the solubility of adenosine by about 100 times, which is extremely significant compared with traditional solvent systems or solubilization methods. The high solubility allows the concentration of adenosine in drug formulations to be greatly increased, providing the possibility for developing highly efficient drug dosage forms. (2) Improves bioavailability: The significant increase in solubility directly promotes the dissolution and absorption process of adenosine in vivo, enabling the drug to reach the target site faster and more effectively, greatly improving the bioavailability of adenosine. Experiments show that after using the ternary eutectic solvent of this invention, the peak blood concentration of adenosine in animals is significantly increased, the duration of drug action is prolonged, and the efficacy of the drug is significantly improved. (3) Simple preparation process: The preparation method of the ternary eutectic solvent provided by this invention only requires simple heating and stirring, without the need for complex equipment and technology. It is easy to operate, easy to mass-produce, reduces production costs, and is conducive to industrial promotion and application. Attached Figure Description
[0020] Figure 1 This is a schematic diagram of the adenosine molecule structure in this invention.
[0021] Figure 2 The malic acid-betaine-adenosine ternary DES nuclear magnetic resonance hydrogen spectrum obtained in Experimental Example 2 of this invention ( 1 H-NMR spectrum.
[0022] Figure 3 This is a comparison of the infrared spectra of malic acid, betaine, adenosine, malic acid-betaine binary DES, and malic acid-betaine-adenosine ternary DES in Experimental Example 2 of this invention.
[0023] Figure 4 Infrared spectra of the malic acid betaine adenosine ternary DES prepared in Example 2 of this invention diluted with water at different ratios.
[0024] Figure 5 It is the negative control group containing only complete culture medium in the live and dead cell staining experiment in Experiment Example 2 of this invention.
[0025] Figure 6 This is the experimental group of the ternary DES sample with a concentration of 2 mg / ml malate-betaine-adenosine in the live-dead cell staining experiment of Experiment Example 2 of the present invention.
[0026] Figure 7 This is the optimized structure diagram of the malic acid betaine adenosine ternary DES of Experimental Example 2 of the present invention.
[0027] Figure 8 The tartrate-betaine-adenosine ternary DES nuclear magnetic resonance hydrogen spectrum of the tartrate-betaine-adenosine trivalent obtained in Experimental Example 4 of this invention ( 1 H-NMR spectrum.
[0028] Figure 9 This is a comparison of the infrared spectra of tartaric acid, betaine, adenosine, tartaric acid-betaine binary DES, and tartaric acid-betaine-adenosine ternary DES in Experimental Example 4 of this invention.
[0029] Figure 10 Infrared spectra of the tartrate betaine adenosine ternary DES diluted with water at different ratios in Example 4 of this invention.
[0030] Figure 11 This is the optimized structure diagram of the tartrate betaine adenosine ternary DES of Experimental Example 4 of the present invention. Detailed Implementation
[0031] This invention provides a ternary eutectic solvent, its preparation method, and its application. To make the objectives, technical solutions, and effects of this invention clearer and more explicit, the invention is further described in detail below. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
[0032] Eutectic solvents (DES), as a novel type of green solvent, have attracted widespread attention in the pharmaceutical formulation field in recent years. DES are typically formed by the interaction of hydrogen bond acceptors (quaternary ammonium salts) and hydrogen bond donors (such as organic acids, polyols, and amides) through hydrogen bonding, exhibiting advantages such as low volatility, high stability, and biodegradability. However, traditional binary DES still have limitations in improving the solubility of certain drugs. For poorly soluble drugs like adenosine, a single binary DES is insufficient to significantly enhance its solubility. Therefore, developing novel and efficient methods and compositions to improve the solubility and bioavailability of adenosine has significant practical and clinical application value.
[0033] Furthermore, compared to solid cocrystal drugs, DES can significantly improve drug solubility. Many solid drugs have low solubility in water or other common solvents, while the unique intermolecular interactions and structural characteristics of DES give it excellent solubility for a variety of drugs, enabling them to be uniformly dispersed in the system at the molecular level, which helps improve drug bioavailability. DES generally has good chemical and thermal stability. Under normal temperature and conventional storage conditions, drugs in DES can maintain stable chemical structures and properties, and are not prone to degradation, deterioration, or crystal form transformation, which is beneficial for long-term drug storage and quality control. By changing the composition and ratio of DES, its physicochemical properties, such as viscosity, density, and surface tension, can be flexibly adjusted to meet different drug delivery needs. For example, for cases requiring local administration, the viscosity of DES can be adjusted to make it easier to apply and adhere to the administration site. DES can achieve stable drug storage and controlled release while reducing the toxic side effects of drugs on normal tissues.
[0034] Based on this, embodiments of the present invention provide a ternary eutectic solvent, which is prepared from component A, component B and component C, wherein component A is a hydrogen bond donor, component B is a hydrogen bond acceptor and component C is adenosine.
[0035] In the ternary DES of this invention, the hydrogen bond acceptor has good hydrophilicity and cationic properties. The carboxyl and hydroxyl groups of the hydrogen bond donor can form strong hydrogen bond interactions with the hydrogen bond acceptor, constructing the framework structure of the binary DES. The active groups such as hydroxyl and amino groups in the adenosine molecule (see schematic diagram) Figure 1As shown, it can further interact with hydrogen bond acceptors and hydrogen bond donors through hydrogen bonds and van der Waals forces to form a stable ternary DES system. This unique intermolecular interaction mode breaks the original strong intermolecular forces between adenosine molecules, allowing it to be uniformly dispersed in the ternary DES system as single molecules or small aggregates, thereby significantly improving the solubility of adenosine. Simultaneously, the presence of ternary DES alters the drug's dissolution and release behavior in vivo, promoting drug absorption and greatly improving the bioavailability of adenosine.
[0036] In some embodiments, component A is selected from one or more of malic acid, tartaric acid, citric acid, and salicylic acid.
[0037] In some embodiments, component B is selected from one or both of L-carnitine and betaine.
[0038] The raw materials used in the embodiments of this invention, namely malic acid, tartaric acid, citric acid, salicylic acid, L-carnitine, and betaine, are all naturally occurring and biodegradable substances. No toxic or harmful organic solvents are required in the preparation process, which is in line with the concept of green chemistry and is environmentally friendly.
[0039] In some embodiments, the molar ratio of component A, component B and component C in the ternary eutectic solvent is (5-15):(5-15):1.
[0040] This ratio can significantly lower the eutectic point of the mixed system (usually lower than the melting point of each individual component) through the synergistic effect of hydrogen bonds, ionic bonds, or van der Waals forces between components, forming a stable eutectic phase. For example, when the proportion of component C is low, an appropriate amount of components A / B can act as a "structural framework," forming a dense network of hydrogen bonds with component C, thereby minimizing the system's free energy and achieving a lower melting temperature.
[0041] In some preferred embodiments, the molar ratio of component A, component B and component C in the ternary eutectic solvent is (8-12):(8-12):1.
[0042] This invention also provides a method for preparing the above-mentioned ternary eutectic solvent, the method comprising the following steps:
[0043] Component A and component B are mixed and subjected to a first heating and stirring process to obtain a mixed solution;
[0044] Component C is added to the above mixed solution, and a second heating and stirring treatment is performed to obtain the ternary eutectic solvent.
[0045] The preparation method of this invention can be completed simply by heating and stirring, without the need for complex equipment and technology. It is easy to operate, easy to mass-produce, reduces production costs, and is conducive to industrial application.
[0046] In some embodiments, the temperature of the first heating and stirring treatment is 50-80°C, and the time is 1-3 hours.
[0047] In some embodiments, the temperature of the second heating and stirring treatment is 35-90°C, and the time is 3-6 hours.
[0048] Setting an upper temperature limit can prevent the thermal decomposition of most organic components, while controlling the lower time limit can ensure that the main reaction proceeds fully and avoid side reactions caused by excessive time.
[0049] In some preferred embodiments, the temperature of the first heating and stirring treatment is 70°C and the time is 1.5 hours.
[0050] In some preferred embodiments, the temperature of the second heating and stirring treatment is 80°C and the time is 3 hours.
[0051] This invention provides the application of the above-described ternary eutectic solvent or the ternary eutectic solvent prepared by the above-described preparation method in the preparation of drugs.
[0052] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are merely some embodiments of the present invention, not all embodiments, and are intended only to illustrate the present invention and not to limit it. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0053] Example 1
[0054] The preparation of malate-L-carnitine-adenosine trivalent DES includes the following steps:
[0055] Step 1: Accurately weigh 1 mol of malic acid (134 g / mol × 1 mol = 134 g) and 1 mol of L-carnitine (161.2 g / mol × 1 mol = 161.2 g), and add them to a three-necked flask equipped with a stirrer and a thermometer.
[0056] Step 2: Place the three-necked flask in a constant temperature water bath, heat to 60°C, stir and react for 2 hours to obtain a homogeneous liquid, namely malic acid L-carnitine binary DES.
[0057] Step 3: Add 0.33 mol of adenosine (267.25 g / mol × 0.33 mol = 88.19 g) to the above binary DES, and continue to stir the reaction at 60 °C for 4 hours to obtain a clear and transparent malic acid-L-carnitine-adenosine ternary DES system.
[0058] Example 2
[0059] The preparation of malic acid-betaine-adenosine ternary DES includes the following steps:
[0060] Step 1: Accurately weigh 1 mol of malic acid (134 g / mol × 1 mol = 134 g) and 1 mol of betaine (117.15 g / mol × 1 mol = 117.15 g), and add them to a three-necked flask equipped with a stirrer and a thermometer.
[0061] Step 2: Place the three-necked flask in a constant temperature water bath, heat to 60°C, and stir for 2 hours to obtain a homogeneous liquid, namely malic acid betaine binary DES.
[0062] Step 3: Add 0.33 mol of adenosine (267.25 g / mol × 0.33 mol = 88.19 g) to the above binary DES, and continue to stir the reaction at 60 °C for 4 hours to obtain a clear and transparent malic acid betaine adenosine ternary DES system.
[0063] The malic betaine binary DES and malic betaine adenosine ternary DES obtained in Example 2 were subjected to 1H NMR spectroscopy. 1 Characterization by H-NMR, NMR spectra such as Figure 2 As shown. Figure 2 The 1H NMR spectra show that, using D₂O as the test solvent, adenosine with two hydrogen atoms on the N-containing five-membered and six-membered rings was clearly identified in the 1H NMR spectrum, with no obvious impurity peaks observed. Infrared characterization was performed on malic acid, betaine, adenosine, and the obtained malic acid-betaine binary DES and malic acid-betaine-adenosine ternary DES: the infrared spectra are shown below. Figure 3 As shown, the NH peak of adenosine shifted, indicating that the amino group of adenosine participated in hydrogen bond formation. Furthermore, compared to the binary DES formed by malic acid and betaine, the ternary DES peak was observed in the 3200-3600 cm⁻¹ range. -1 and 1400-1800cm -1 The peak shape and position changed. This proves that adenosine and malate betaine formed a ternary DES through several interatomic hydrogen bonding interactions.
[0064] To investigate the dilution stability of the ternary eutectic solvent (DES) product prepared in Example 2, a series of gradient dilution strategies were employed for precise processing. During the experiment, based on preliminary experimental results and product properties, suitable diluents were selected, and high-precision instruments such as pipettes and volumetric flasks were used to prepare product solutions of different concentration gradients according to strict dilution ratios (e.g., mass ratios of 1:3, 1:4, 1:5, etc.). Subsequently, Fourier transform infrared spectroscopy was performed on the diluted products of different concentrations: the infrared spectra are shown below. Figure 4 As shown, the changes in the position, intensity, and shape of infrared absorption peaks at different dilution factors were analyzed to determine the alterations in intermolecular interactions. The infrared spectra of the diluted samples show that with increasing water content in DES, both the intensity and width of the OH absorption peak at 3200-3600 nm increase, while the carbonyl absorption peak gradually disappears. Simultaneously, the absorption peak in the infrared fingerprint region (1300-400 nm) decreases in intensity with the addition of water. This indicates that the addition of water leads to the gradual breakdown of the hydrogen bond network in DES.
[0065] To investigate the cytotoxicity of the ternary eutectic solvent (DES) product prepared in Example 2, a live-dead cell staining method was used to evaluate the toxicity of L929 mouse fibroblasts. L929 cells were stained at 5 × 10⁻⁶ cells per cell line. 4 Cells were seeded at a density of 24 wells and pre-cultured at 37°C in a 5% CO2 incubator for 24 h. The experimental groups were supplemented with a ternary DES sample solution to a final concentration of 2 mg / ml, while a negative control group containing only complete culture medium was set up. After culturing for another 24 h, the culture medium in the wells was discarded, and the cells were gently washed three times with PBS. A staining working solution containing 2 μM calcein and 4 μM propidium iodide was added, and the cells were incubated at 37°C in the dark for 20 min. Cell images were observed and captured using a fluorescence microscope, and five fields of view were randomly selected from each sample for analysis. Figure 5 This serves as a negative control group containing only complete culture medium. Figure 6 The experimental group consisted of a ternary DES sample with a concentration of 2 mg / ml of malic acid, betaine, and adenosine. Figure 7 Optimized structure diagram of the ternary DES of malic acid betaine adenosine.
[0066] The results showed that after staining, the live cells in the experimental group exhibited bright green fluorescence, and the proportion of live cells was not significantly different from that in the negative control group (p>0.05). This indicates that the ternary DES sample at this concentration had no significant toxic effect on L929 cells and possessed good biocompatibility.
[0067] Example 3
[0068] The preparation of L-carnitine adenosine ternary DES by tartrate includes the following steps:
[0069] Step 1: Accurately weigh 1 mol of tartaric acid (150.09 g / mol × 1 mol = 150.09 g) and 1 mol of L-carnitine (161.2 g / mol × 1 mol = 161.2 g), and add them to a three-necked flask equipped with a stirrer and a thermometer.
[0070] Step 2: Place the three-necked flask in a constant temperature water bath, heat to 60°C, stir and react for 2 hours to obtain a homogeneous liquid, namely tartrate betaine binary DES.
[0071] Step 3: Add 0.33 mol of adenosine (267.25 g / mol × 0.33 mol = 88.19 g) to the above binary DES, and continue to stir the reaction at 60 °C for 4 hours to obtain a clear and transparent tartrate betaine adenosine ternary DES system.
[0072] Example 4
[0073] The preparation of tartrate-betaine-adenosine ternary DES includes the following steps:
[0074] Step 1: Accurately weigh 1 mol of tartaric acid (150.09 g / mol × 1 mol = 150.09 g) and 1 mol of betaine (117.15 g / mol × 1 mol = 117.15 g), and add them to a three-necked flask equipped with a stirrer and a thermometer.
[0075] Step 2: Place the three-necked flask in a constant temperature water bath, heat to 60°C, stir and react for 2 hours to obtain a homogeneous liquid, namely tartrate betaine binary DES.
[0076] Step 3: Add 0.33 mol of adenosine (267.25 g / mol × 0.33 mol = 88.19 g) to the above binary DES, and continue to stir the reaction at 60 °C for 4 hours to obtain a clear and transparent tartrate betaine adenosine ternary DES system.
[0077] The 1H NMR spectra of the tartrate betaine binary DES and the tartrate betaine adenosine ternary DES obtained in Example 4 were analyzed. 1 Characterization by H-NMR, NMR spectra such as Figure 8 As shown. Figure 8 The 1H NMR spectra showed that, using D₂O as the test solvent, adenosine with two hydrogen atoms on both the N-containing five-membered and six-membered rings was clearly visible in the 1H NMR spectrum, with no obvious impurity peaks observed. Infrared characterization was performed on tartaric acid, betaine, adenosine, and the obtained tartaric acid-betaine binary DES and tartaric acid-betaine-adenosine ternary DES. The infrared spectra are shown below. Figure 9As shown, the NH peak of adenosine shifted, indicating that the amino group of adenosine participated in hydrogen bond formation. Furthermore, compared to the binary DES formed by tartaric acid and betaine, the ternary DES showed a shift in the 3200-3600 cm⁻¹ peak. -1 and 1400-1800cm -1 The peak shape and position changed. This proves that adenosine and tartrate betaine formed a ternary DES through several interatomic hydrogen bonding interactions.
[0078] Figure 10 The images show the infrared spectra of tartrate betaine adenosine ternary DES diluted with water at different ratios. The FT-IR spectra of the diluted samples show that with increasing water content in the DES, both the intensity and width of the OH absorption peak at 3200-3600 nm increase, while the carbonyl absorption peak gradually disappears. Simultaneously, the intensity of the absorption peak in the infrared fingerprint region (1300-400 nm) decreases with the addition of water. This indicates that the addition of water leads to the gradual breakdown of the hydrogen bond network in the DES.
[0079] Figure 11 Optimized structure diagram of tartrate betaine adenosine ternary DES.
[0080] Adenosine solubilization effect test
[0081] Take 10g of each of the ternary DES compositions prepared in Example 2 and Example 4, and place them in two 50mL volumetric flasks. Slowly add deionized water to the volumetric flasks while stirring continuously until the volumetric flask marks are reached, thus preparing an aqueous solution containing ternary DES.
[0082] Take another 50mL volumetric flask, add 10g of adenosine, then add deionized water and stir until the volumetric flask reaches the mark, and use it as the control group.
[0083] The three volumetric flasks were placed in a constant temperature water bath shaker at 25°C and shaken for 24 hours to allow the adenosine to dissolve completely. After shaking, the volumetric flasks were removed and filtered through a 0.45 μm microporous membrane. The concentration of adenosine in the filtrate was determined by high performance liquid chromatography (HPLC).
[0084] The solubility of adenosine in the control group was 0.5 mg / mL, the solubility of adenosine in the malic acid betaine adenosine ternary DES system in Example 2 was 50 mg / mL, and the solubility of adenosine in the tartrate betaine adenosine ternary DES system in Example 4 was 48 mg / mL. Compared with the control group, the solubility increased by about 100 times.
[0085] Surfactant solubilization effect test of adenosine
[0086] Prepare a 0.5% sodium dodecyl sulfate (SDS) aqueous solution. Add 10g of adenosine to 50mL of the SDS aqueous solution and determine the solubility of adenosine in the solution.
[0087] The solubility of adenosine in this system was measured to be 8 mg / mL, which is far lower than the solubility enhancement effect of the ternary DES provided in the embodiments of the present invention.
[0088] It should be understood that the application of the present invention is not limited to the examples above. Those skilled in the art can make improvements or modifications based on the above description, and all such improvements and modifications should fall within the protection scope of the appended claims.
Claims
1. A ternary eutectic solvent, characterized in that, The ternary eutectic solvent is prepared from components A, B and C, wherein component A is a hydrogen bond donor, component B is a hydrogen bond acceptor and component C is adenosine. Component A is selected from malic acid or tartaric acid; Component B is selected from betaine; In the ternary eutectic solvent, the molar ratio of component A, component B and component C is 1:1:0.
33.
2. A method for preparing the ternary eutectic solvent according to claim 1, characterized in that, The preparation method includes the following steps: Component A and component B are mixed and subjected to a first heating and stirring process to obtain a mixed solution; Component C is added to the above mixed solution, and a second heating and stirring treatment is performed to obtain the ternary eutectic solvent.
3. The method for preparing the ternary eutectic solvent according to claim 2, characterized in that, The temperature of the first heating and stirring treatment is 50-80℃, and the time is 1-3 hours.
4. The method for preparing the ternary eutectic solvent according to claim 2, characterized in that, The second heating and stirring treatment is carried out at a temperature of 35-90℃ for 3-6 hours.