Reduced coenzyme Q10 / gamma-aminobutyric acid eutectic crystal and preparation method thereof

Abstract

The invention belongs to the technical field of coenzyme Q10 products, and particularly relates to a reduced coenzyme Q10 / gamma-aminobutyric acid eutectic crystal and a preparation method thereof. The reduced coenzyme Q10 / gamma-aminobutyric acid eutectic crystal is a eutectic crystal formed by reduced coenzyme Q10 and gamma-aminobutyric acid. The reduced coenzyme Q10 / gamma-aminobutyric acid eutectic provided by the invention takes gamma-aminobutyric acid as a eutectic ligand of the reduced coenzyme Q10, not only can the melting point be increased to 70 DEG C or above, but also the reduced coenzyme Q10 / gamma-aminobutyric acid eutectic has good physical and chemical stability, and also has relatively high bioavailability. Besides, the preparation method of the reduced coenzyme Q10 / gamma-aminobutyric acid eutectic crystal provided by the invention is simple, good in repeatability, strong in operability and beneficial to large-scale industrial production, and has an important value for developing a reduced coenzyme Q10 solid oral preparation in the future.

Description

Technical Field

[0001] This invention belongs to the technical field of coenzyme Q10 products, specifically relating to a reduced coenzyme Q10 / γ-aminobutyric acid cocrystal and its preparation method. Background Technology

[0002] Coenzyme Q10 (CoQ10) is a fat-soluble compound widely found in the mitochondria of human cells and is a core substance for energy metabolism and antioxidant defense. As an indispensable substance in human cells, CoQ10 plays a crucial role in energy metabolism and antioxidant defense. As a key component of the mitochondrial electron transport chain, CoQ10 directly participates in the synthesis of adenosine triphosphate (ATP), providing energy for high-energy-consuming organs such as the heart and brain. CoQ10 can neutralize free radicals, protect cell membranes, lipoproteins, and DNA from oxidative damage, delay cell aging, improve cardiomyocyte function, and help maintain blood pressure and vascular elasticity. CoQ10 can be used as an adjunct to medication for cardiovascular diseases, and has a certain effect on improving heart failure, alleviating hypertension, managing diabetes, and preventing neurodegenerative diseases. As a food supplement, CoQ10 not only has antioxidant and anti-aging properties but also combats fatigue and improves athletic performance. Currently, CoQ10 is widely used in the food, health product, cosmetic, and pharmaceutical industries and is highly favored by scholars and consumers.

[0003] Coenzyme Q10 exists in two forms: oxidized and reduced. Reduced coenzyme Q10, commonly known as coenzyme QH, is the main form found in the human bloodstream and directly participates in energy metabolism and antioxidant processes. Compared to oxidized coenzyme Q10, coenzyme QH has better antioxidant properties and more efficient absorption. However, coenzyme QH is chemically extremely unstable, with a low melting point, poor heat resistance, and high sensitivity to oxygen and light. During production and transportation, coenzyme QH is easily oxidized and degraded, resulting in poor stability and severely affecting its quality.

[0004] Furthermore, coenzyme QH may be prematurely oxidized in the gastrointestinal environment, significantly reducing its actual absorption efficiency. The core issue lies in its insufficient crystal stability, which affects its solubility and absorption in the gastrointestinal tract. Oxidized coenzyme Q10, due to its poor hydrophilicity and high lattice energy, is difficult to dissolve in gastrointestinal fluids, limiting its bioavailability. Although coenzyme QH is theoretically more easily absorbed than oxidized coenzyme Q10, its absorption rate is difficult to reach ideal levels due to stability issues, resulting in poor bioavailability. Summary of the Invention

[0005] The primary objective of this invention is to overcome the shortcomings of coenzyme QH, such as low melting point, poor stability, and low bioavailability, and to provide a reduced coenzyme Q10 / γ-aminobutyric acid cocrystal with high melting point, high stability, and high bioavailability.

[0006] The second objective of this invention is to provide a method for preparing the above-mentioned reduced coenzyme Q10 / γ-aminobutyric acid cocrystal.

[0007] A third objective of this invention is to provide a coenzyme Q10 product containing the aforementioned reduced coenzyme Q10 / γ-aminobutyric acid cocrystal.

[0008] The reduced coenzyme Q10 / γ-aminobutyric acid cocrystal (QH-GABA-CCIII) provided by the present invention is a cocrystal formed by reduced coenzyme Q10 (coenzyme QH) and γ-aminobutyric acid (GABA).

[0009] The method for preparing reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by the present invention includes recrystallizing reduced coenzyme Q10 and γ-aminobutyric acid together in a solvent to obtain reduced coenzyme Q10 / γ-aminobutyric acid cocrystal, or grinding reduced coenzyme Q10 and γ-aminobutyric acid to obtain reduced coenzyme Q10 / γ-aminobutyric acid cocrystal.

[0010] During the research and development process, this invention surprisingly discovered that using γ-aminobutyric acid (GABA) as the co-crystal ligand for reduced coenzyme Q10 results in a reduced coenzyme Q10 / γ-aminobutyric acid co-crystal that not only possesses a melting point above 70°C but also exhibits excellent physical and chemical stability, along with high bioavailability. Furthermore, the preparation method for the reduced coenzyme Q10 / γ-aminobutyric acid co-crystal provided by this invention is simple, reproducible, and highly operable, facilitating large-scale industrial production and holding significant value for the future development of solid oral formulations of reduced coenzyme Q10. Attached Figure Description

[0011] Figure 1 The X-ray powder diffraction (XRPD) pattern of coenzyme QH obtained in the preparation example; Figure 2 Differential scanning calorimetry (DSC) spectra of coenzyme QH obtained in the preparation example; Figure 3 The XRPD pattern of the eutectic QH-GABA-CCIII-1 obtained in Example 1; Figure 4 The XRPD patterns of the cocrystal QH-GABA-CCIII-1 obtained in Example 1 and the coenzyme QH and GABA obtained in the preparation example are overlaid. Figure 5The DSC spectrum of the eutectic QH-GABA-CCIII-1 obtained in Example 1; Figure 6 Thermogravimetric analysis (TGA) spectrum of the eutectic QH-GABA-CCIII-1 obtained in Example 1; Figure 7 The XRPD pattern of the eutectic QH-GABA-CCIII-2 obtained in Example 2; Figure 8 The DSC spectrum of the eutectic QH-GABA-CCIII-2 obtained in Example 2; Figure 9 The XRPD pattern of the eutectic QH-GABA-CCIII-3 obtained in Example 3; Figure 10 The DSC spectrum of the eutectic QH-GABA-CCIII-3 obtained in Example 3; Figure 11 The XRPD pattern of the eutectic QH-GABA-CCIII-4 obtained in Example 4; Figure 12 The DSC spectrum of the eutectic QH-GABA-CCIII-4 obtained in Example 4; Figure 13 The XRPD pattern of the eutectic QH-GABA-CCIII-5 obtained in Example 5; Figure 14 The DSC spectrum of the eutectic QH-GABA-CCIII-5 obtained in Example 5; Figure 15 The XRPD pattern of the mixture of cocrystal QH-GABA-CCIII-6 and coenzyme QH obtained in Example 6; Figure 16 The DSC spectrum of the mixture of cocrystal QH-GABA-CCIII-6 and coenzyme QH obtained in Example 6; Figure 17 The graph shows the color and shape changes of the cocrystal QH-GABA-CCIII-1 obtained in Example 1 and the coenzyme QH obtained in the preparation example. Figure 18 The particle size distribution diagram of coenzyme QH obtained in the preparation example; Figure 19 The particle size distribution diagram of the eutectic QH-GABA-CCIII-1 obtained in Example 1 is shown. Figure 20 This is a comparison chart of the tableting effects of the eutectic QH-GABA-CCIII-1 obtained in Example 1 and the eutectic QH-Nam-CCIII obtained in Comparative Example 1. Detailed Implementation

[0012] The reduced coenzyme Q10 / γ-aminobutyric acid (GABA) cocrystal provided by this invention is a cocrystal formed by reduced coenzyme Q10 and γ-aminobutyric acid (GABA). The stoichiometric ratio of reduced coenzyme Q10 to GABA in the cocrystal is particularly preferably 1:(0.5-2.5), at which point the resulting reduced coenzyme Q10 / γ-aminobutyric acid cocrystal exhibits higher stability and bioavailability. Specifically, the stoichiometric ratio of reduced coenzyme Q10 to GABA in the cocrystal can be 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.05, 1:1.1, 1:1.15, 1:1.2, 1:1.25, 1:1.3, 1:1.35, 1:1.4, 1:1.45, 1:1. :1.5, 1:1.55, 1:1.6, 1:1.65, 1:1.7, 1:1.75, 1:1.8, 1:1.85, 1:1.9, 1:1.95, 1:2, 1:2.05, 1:2.1, 1:2.15, 1:2.2, 1:2.25, 1:2.3, 1:2.35, 1:2.4, 1:2.45, 1:2.5 or any value between them.

[0013] In a preferred embodiment, the X-ray powder diffraction pattern of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal obtained by Cu-Kα radiation has characteristic peaks at at least the following 2θ angles: 2.9°, 4.3°, 8.6°, 10.0°, 11.5°, 15.7°, 17.2°, 18.7°, 23.3°, and 24.1°, with an error of ±0.2° for the 2θ angle.

[0014] In a preferred embodiment, the differential scanning calorimetry (DSC) spectrum of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal has a characteristic endothermic peak at least at 73℃±2℃, specifically at 71℃, 71.2℃, 71.4℃, 71.6℃, 71.8℃, 72℃, 72.2℃, 72.4℃, 72.6℃, 72.8℃, 73℃, 73.2℃, 73.4℃, 73.6℃, ​​73.8℃, 74℃, 74.2℃, 74.4℃, 74.6℃, 74.8℃, 75℃, or any value between them.

[0015] In a preferred embodiment, the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal exhibits a unimodal particle size distribution, with a particle size ranging from 0.01 μm to 20 μm, such as 0.01 μm, 0.03 μm, 0.05 μm, 0.08 μm, 0.1 μm, 0.15 μm, 0.2 μm, 0.25 μm, 0.3 μm, 0.35 μm, 0.4 μm, 0.45 μm, 0.5 μm, 0.55 μm, and 0. 6μm, 0.65μm, 0.7μm, 0.75μm, 0.8μm, 0.85μm, 0.9μm, 0.95μm, 1μm, 2μm, 3μm, 4μm, 5μm, 6μm, 7μm, 8μm, 9μm, 10μm, 11μm, 12μm, 13μm, 14μm, 15μm, 16μm, 17μm, 18μm, 19μm, 20μm or any value between them.

[0016] The method for preparing reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by this invention includes: Method 1: Recrystallize reduced coenzyme Q10 and γ-aminobutyric acid (GABA) together in a solvent to obtain a reduced coenzyme Q10 / γ-aminobutyric acid cocrystal; or, Method 2: Reduced coenzyme Q10 and γ-aminobutyric acid are ground to obtain reduced coenzyme Q10 / γ-aminobutyric acid cocrystal.

[0017] The method for preparing reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by this invention is simple to operate, the crystallization process is easy to control, the crystallinity is high, and the reproducibility is good, and the cocrystal of coenzyme Q10 and GABA can be stably obtained.

[0018] In the preparation process of the above-mentioned reduced coenzyme Q10 / γ-aminobutyric acid cocrystal, the preferred molar ratio of reduced coenzyme Q10 to γ-aminobutyric acid is 1:(0.5-2.5). At this ratio, the resulting reduced coenzyme Q10 / γ-aminobutyric acid cocrystal exhibits higher stability and bioavailability. Specifically, the molar ratio of reduced coenzyme Q10 to γ-aminobutyric acid can be 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.05, 1:1.1, 1:1.15, 1:1.2, 1:1.25, 1:1.3, 1:1.35, 1:1.4, 1:1.45, 1:1.5, or 1:1. 0.55, 1:1.6, 1:1.65, 1:1.7, 1:1.75, 1:1.8, 1:1.85, 1:1.9, 1:1.95, 1:2, 1:2.05, 1:2.1, 1:2.15, 1:2.2, 1:2.25, 1:2.3, 1:2.35, 1:2.4, 1:2.45, 1:2.5 or any value between them.

[0019] In the preparation process of the above-mentioned reduced coenzyme Q10 / γ-aminobutyric acid cocrystal, the solvent can be any existing inert liquid medium that has a certain solubility for reduced coenzyme Q10 and γ-aminobutyric acid and will not cause deterioration of reduced coenzyme Q10 and γ-aminobutyric acid. Specifically, it can be selected from at least one of water, alcohol solvents, nitrile solvents, ketone solvents, ester solvents, alkane solvents, aromatic hydrocarbon solvents, and haloalkane solvents. Examples of alcohol solvents include at least one of methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, and tert-butanol. Examples of nitrile solvents include at least one of acetonitrile, butyronitrile, valerate, and methoxypropionitrile. Examples of ketone solvents include at least one of acetone, butanone, cyclohexanone, methylcyclohexanone, and isophorone. The ester solvent may include at least one of ethyl formate, propyl formate, isopropyl formate, butyl formate, isobutyl formate, isoamyl formate, ethyl acetate, n-propyl acetate, and isopropyl acetate. The alkane solvent may include at least one of n-pentane, n-hexane, n-heptane, n-octane, isopentane, isohexane, 2,2,4-trimethylpentane, isododecane, cyclohexane, and methylcyclohexane. The aromatic hydrocarbon solvent may include at least one of toluene, xylene, trimethylbenzene, ethylbenzene, and chlorobenzene. The haloalkane solvent may include at least one of dichloromethane, trichloromethane, 1,2-dichloroethane, and trichloroethane. The solvent is preferably at least one of alcohol solvents, nitrile solvents, ester solvents, and alkane solvents, more preferably ethanol, acetonitrile, ethyl acetate, and mixtures thereof with other solvents, wherein the other solvents are selected from at least one of methanol, acetonitrile, and n-hexane. Using this preferred solvent has the advantages of low cost and recyclability.

[0020] In the preparation of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal described above, the recrystallization method preferably includes dissolving reduced coenzyme Q10 and γ-aminobutyric acid together in a solvent by stirring and then stirring to induce crystallization or allowing the mixture to stand and evaporate to induce crystallization. The stirring and dissolving conditions preferably include a temperature of 20℃-80℃, such as 20℃, 25℃, 30℃, 35℃, 40℃, 45℃, 50℃, 55℃, 60℃, 65℃, 70℃, 75℃, 80℃, or any value between these values; and a time of 0.1h-2h, such as 0.1h, 0.2h, 0.4h, 0.6h, 0.8h, 1h, 1.2h, 1.4h, 1.6h, 1.8h, 2h, or any value between these values. The preferred conditions for stirring, cooling, and crystallization include a temperature of 4℃-40℃, such as 4℃, 6℃, 8℃, 10℃, 12℃, 15℃, 18℃, 20℃, 22℃, 25℃, 28℃, 30℃, 32℃, 35℃, 38℃, 40℃, or any value between them; and a time of 1h-48h, such as 1h, 2h, 4h, 6h, 8h, 10h, 12h, 15h, 18h, 20h, 22h, 25h, 28h, 30h, 32h, 35h, 38h, 40h, 42h, 45h, 48h, or any value between them. The conditions for static evaporation and crystallization include a temperature of 4℃-40℃, such as 4℃, 6℃, 8℃, 10℃, 12℃, 15℃, 18℃, 20℃, 22℃, 25℃, 28℃, 30℃, 32℃, 35℃, 38℃, 40℃, or any value between them; and a time of 1 day-30 days, such as 1 day, 5 days, 10 days, 15 days, 20 days, 25 days, 30 days, or any value between them. Furthermore, after recrystallization is complete, the solvent usually needs to be removed. The solvent removal method can be filtration, and the resulting product can be dried after filtration. In addition, the grinding can be dry grinding or wet grinding.

[0021] The present invention also provides a coenzyme Q10 product, wherein the coenzyme Q10 product contains the above-mentioned reduced coenzyme Q10 / γ-aminobutyric acid cocrystal.

[0022] The coenzyme Q10 product provided by this invention contains reduced coenzyme Q10 / γ-aminobutyric acid (GABA) cocrystals. Specific examples of the coenzyme Q10 product include, but are not limited to, food, cosmetics, pharmaceuticals, and feed. In addition to the reduced coenzyme Q10 / γ-aminobutyric acid cocrystals, the coenzyme Q10 product also contains other acceptable raw materials. These other acceptable raw materials are formulated according to the desired product. For example, food may contain main food ingredients and food-acceptable edible food additives, such as sweeteners, flavoring agents, preservatives, fragrances, and colorings; cosmetics may contain cosmetic-acceptable main cosmetic ingredients and additives, such as solvents, fragrances, preservatives, flavorings, and colorings; pharmaceuticals may contain pharmaceutically active ingredients and pharmaceutically acceptable excipients, such as carriers, diluents, adjuvants, and colorings; feed may contain feed main ingredients, such as soybean meal and hay, and feed-acceptable feed excipients, such as sweeteners, flavoring agents, preservatives, fragrances, and colorings, but this invention is not limited to these.

[0023] The analytical methods and conditions involved in the following examples and comparative examples are as follows: (1) Detection and analysis of coenzyme QH content: Instrument: Agilent Technologies 1260 Infinity (with UV detector); Chromatographic column: ZORBX SB-C18, 4.6mm×100mm, 3.5μm column; Column temperature: 35℃; Detection wavelength: 290nm; Eluent: A mixture of acetonitrile and ethanol in a 1:1 volume ratio; Elution rate: 1 mL / min.

[0024] (2) GABA content detection and analysis: Instrument: Agilent Technologies 1260 Infinity (with UV detector); Chromatographic column: HALO COLUMNS C18, 4.6 mm × 100 mm, 2.7 μm column; Column temperature: 25℃; Detection wavelength: 360nm; Eluent: Phase A is pH 6.5 0.1M sodium acetate aqueous solution-acetonitrile (volume ratio 93:7), Phase B is water-acetonitrile (volume ratio 20:80), and the volume ratio of Phase A to Phase B is 50:50; Elution rate: 1 mL / min.

[0025] (3) Differential scanning calorimetry: Instrument model: DSC Q2000; The detection method involves heating to 250℃ at a rate of 10℃ / min.

[0026] (4) Thermogravimetric analysis: Instrument model: TGA 500; The detection method involves heating to 250℃ at a rate of 10℃ / min.

[0027] (5) XRPD analysis: Instrument model: D2 PHASER BRUKER; Target: Cu-Kα (30KV, 10mA); Scan range: 3°-40° (2θ value); Scan step size: 0.1s.

[0028] (6) HPLC-MS / MS analysis: Instrument models: ACQUITY UPLC I-Class ultra-high performance liquid chromatograph, Xevo TQ-S triple quadrupole mass spectrometer; Chromatographic column: ACQUITY UPLC-BEH C18 column, 2.1mm×50mm, 1.7μm column; Column temperature: 25℃; Eluent: Methanol (containing 0.1 wt% formic acid); Elution rate: 0.6 mL / min; Mass spectrometry conditions: ESI source, positive mode, scan mode MRM, capillary voltage 2.5kV, cone voltage 20V.

[0029] Preparation Example 200 mL of n-hexane, 20 g of sodium hydrosulfite, and 200 mL of pure water were added to 20 g of oxidized coenzyme Q10. The mixture was stirred and reacted at 30 °C for 1 h. After the reaction was completed, the mixture was allowed to stand and separate into layers. The resulting oil phase was washed three times with water and concentrated to dryness. Then, 300 mL of ethanol was added, and the mixture was cooled and crystallized for 2 h. After filtration, the resulting precipitate was dried in a vacuum drying oven at 25 °C for 12 h to obtain coenzyme QH. The content of the obtained coenzyme QH was determined by high performance liquid chromatography (HPLC), and the results showed that the coenzyme QH content was 99.5%.

[0030] Coenzyme QH was characterized by X-ray powder diffraction (XRPD), and the results are shown in the figure. Figure 1 .like Figure 1As shown, the X-ray powder diffraction (XRPD) pattern of this coenzyme QH exhibits characteristic peaks at 2θ angles of approximately 4.7°±0.2°, 6.2°±0.2°, 14.0°±0.2°, 15.6°±0.2°, 17.1°±0.2°, 18.7°±0.2°, 19.1°±0.2°, 20.3°±0.2°, 23.0°±0.2°, and 27.4°±0.2°. Furthermore, differential scanning calorimetry (DSC) was used to characterize coenzyme QH, and the results are as follows... Figure 2 As shown, it has a characteristic endothermic peak at 52.22℃.

[0031] The obtained coenzyme QH was used as the experimental material for the following examples, comparative examples, and test cases.

[0032] Example 1 This embodiment illustrates the preparation of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by the present invention.

[0033] 0.87 g of coenzyme QH and 0.1 g of GABA were dissolved in 30 mL of methanol and stirred at 60 °C for 0.5 h. After stirring at room temperature (25 °C) for 18 h to crystallize, the resulting white precipitate was filtered and dried in a vacuum drying oven at room temperature (25 °C) for 12 h to obtain the eutectic QH-GABA-CCIII, denoted as QH-GABA-CCIII-1.

[0034] The contents of coenzyme QH and GABA in the obtained cocrystal QH-GABA-CCIII-1 were determined by high performance liquid chromatography. The results showed that the content of coenzyme QH was 89.6% and the content of GABA was 9.9%. Therefore, the stoichiometric ratio of coenzyme QH to GABA in the cocrystal QH-GABA-CCIII-1 was approximately 1:1.

[0035] The eutectic QH-GABA-CCIII-1 was characterized by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). The results are shown in the figures below. Figures 3-6 .

[0036] like Figure 3 As shown, the X-ray powder diffraction pattern of the eutectic QH-GABA-CCIII-1 exhibits characteristic peaks at 2θ angles of 2.9°±0.2°, 4.3°±0.2°, 8.6°±0.2°, 10.0°±0.2°, 11.5°±0.2°, 15.7°±0.2°, 17.2°±0.2°, 18.7°±0.2°, 23.3°±0.2°, and 24.1°±0.2°.

[0037] like Figure 4As shown, the XRPD patterns of this eutectic QH-GABA-CCIII-1 are significantly different from those of coenzyme QH and GABA, and the polymorphisms of coenzyme QH and GABA themselves have been excluded, directly indicating that a new eutectic structure has been formed.

[0038] like Figure 5 As shown, the differential scanning calorimetry spectrum of the eutectic QH-GABA-CCIII-1 has a characteristic endothermic peak at approximately 73.43℃, while the melting point of coenzyme QH itself is around 50℃, indicating that the melting point of the eutectic is significantly higher than that of coenzyme QH.

[0039] like Figure 6 As shown, the thermogravimetric analysis (TGA) spectrum of the eutectic QH-GABA-CCIII-1 showed almost no weight loss before 100℃, indicating that it is a eutectic nonsolvent and does not contain organic solvents.

[0040] Example 2 This embodiment illustrates the preparation of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by the present invention.

[0041] 1.0 g of coenzyme QH and 0.12 g of GABA were dissolved in 40 mL of acetonitrile / ethanol mixed solvent (acetonitrile to ethanol volume ratio of 1:1). The solution was stirred at 60 °C for 0.5 h and then stirred at room temperature (25 °C) for 20 h to crystallize. The resulting white precipitate was filtered and dried in a vacuum drying oven at room temperature (25 °C) for 12 h to obtain the eutectic QH-GABA-CCIII, denoted as QH-GABA-CCIII-2.

[0042] The contents of coenzyme QH and GABA in the obtained cocrystal QH-GABA-CCIII-2 were determined by high performance liquid chromatography. The results showed that the coenzyme QH content was 89.2% and the GABA content was 9.0%. Therefore, the stoichiometric ratio of coenzyme QH to GABA in the cocrystal QH-GABA-CCIII was approximately 1:1.

[0043] The eutectic QH-GABA-CCIII-2 was characterized by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the results are shown in the figures below. Figure 7 and Figure 8 .

[0044] like Figure 7As shown, the X-ray powder diffraction pattern of this eutectic QH-GABA-CCIII-2 also exhibits characteristic peaks at 2θ angles of approximately 2.9°±0.2°, 4.3°±0.2°, 8.6°±0.2°, 10.0°±0.2°, 11.5°±0.2°, 15.7°±0.2°, 17.2°±0.2°, 18.7°±0.2°, 23.3°±0.2°, and 24.1°±0.2°.

[0045] like Figure 8 As shown, the differential scanning calorimetry spectrum of the eutectic QH-GABA-CCIII-2 has a characteristic endothermic peak at approximately 72.06 °C.

[0046] Example 3 This embodiment illustrates the preparation of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by the present invention.

[0047] 5.0 g of coenzyme QH and 0.6 g of GABA were dissolved in 20 mL of a hexane / ethanol mixed solvent (hexane to ethanol volume ratio of 1:2). The mixture was stirred at 60 °C for 20 min and then stirred at room temperature (25 °C) for 15 h to crystallize. The resulting white precipitate was filtered and dried in a vacuum drying oven at room temperature (25 °C) for 12 h to obtain the eutectic QH-GABA-CCIII, denoted as QH-GABA-CCIII-3.

[0048] The contents of coenzyme QH and GABA in the obtained cocrystal QH-GABA-CCIII-3 were determined by high performance liquid chromatography. The results showed that the content of coenzyme QH was about 86.76% and the content of GABA was about 12.98%. Therefore, the stoichiometric ratio of coenzyme QH to GABA in the cocrystal QH-GABA-CCIII-3 was about 1:1.25.

[0049] The eutectic QH-GABA-CCIII-3 was characterized by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the results are shown in the figures below. Figure 9 and Figure 10 .

[0050] like Figure 9 As shown, the X-ray powder diffraction pattern of this eutectic QH-GABA-CCIII-3 also exhibits characteristic peaks at 2θ angles of approximately 2.9°±0.2°, 4.3°±0.2°, 8.6°±0.2°, 10.0°±0.2°, 11.5°±0.2°, 15.7°±0.2°, 17.2°±0.2°, 18.7°±0.2°, 23.3°±0.2°, and 24.1°±0.2°.

[0051] like Figure 10As shown, the differential scanning calorimetry spectrum of the eutectic QH-GABA-CCIII-3 has a characteristic endothermic peak at approximately 72.22 °C.

[0052] Example 4 This embodiment illustrates the preparation of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by the present invention.

[0053] 1.6 g of coenzyme QH and 0.2 g of GABA were dissolved in 50 mL of acetonitrile. The mixture was stirred at 60 °C for 10 min and then stirred at 4 °C for 24 h to crystallize. The resulting white precipitate was filtered and dried in a vacuum drying oven at 25 °C for 12 h to obtain the eutectic QH-GABA-CCIII, denoted as QH-GABA-CCIII-4.

[0054] The contents of coenzyme QH and GABA in the obtained cocrystal QH-GABA-CCIII-4 were determined by high performance liquid chromatography. The results showed that the coenzyme QH content was 90.6% and the GABA content was 10.3%. Therefore, the stoichiometric ratio of coenzyme QH to GABA in the cocrystal QH-GABA-CCIII-4 was approximately 1:1.

[0055] The eutectic QH-GABA-CCIII-4 was characterized by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the results are shown in the figures below. Figure 11 and Figure 12 .

[0056] like Figure 11 As shown, the X-ray powder diffraction pattern of this eutectic QH-GABA-CCIII-4 also exhibits characteristic peaks at 2θ angles of approximately 2.9°±0.2°, 4.3°±0.2°, 8.6°±0.2°, 10.0°±0.2°, 11.5°±0.2°, 15.7°±0.2°, 17.2°±0.2°, 18.7°±0.2°, 23.3°±0.2°, and 24.1°±0.2°.

[0057] like Figure 12 As shown, the differential scanning calorimetry spectrum of the eutectic QH-GABA-CCIII-4 has a characteristic endothermic peak at approximately 73.08 °C.

[0058] Example 5 This embodiment illustrates the preparation of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal provided by the present invention.

[0059] 2.0 g of coenzyme QH and 0.24 g of GABA were dissolved in 50 mL of ethyl acetate and stirred at 60 °C for 0.5 h. After stirring at room temperature (25 °C) for 24 h to crystallize, the resulting white precipitate was filtered and dried in a vacuum drying oven at room temperature (25 °C) for 12 h to obtain the eutectic QH-GABA-CCIII, denoted as QH-GABA-CCIII-5.

[0060] The contents of coenzyme QH and GABA in the obtained cocrystal QH-GABA-CCIII-5 were determined by high performance liquid chromatography. The results showed that the content of coenzyme QH was 89.2% and the content of GABA was 9.7%. Therefore, the stoichiometric ratio of coenzyme QH to GABA in the cocrystal QH-GABA-CCIII-5 was approximately 1:1.

[0061] The eutectic QH-GABA-CCIII-5 was characterized by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the results are shown in the figures below. Figure 13 and Figure 14 .

[0062] like Figure 13 As shown, the X-ray powder diffraction pattern of this eutectic QH-GABA-CCIII-5 also exhibits characteristic peaks at 2θ angles of approximately 2.9°±0.2°, 4.3°±0.2°, 8.6°±0.2°, 10.0°±0.2°, 11.5°±0.2°, 15.7°±0.2°, 17.2°±0.2°, 18.7°±0.2°, 23.3°±0.2°, and 24.1°±0.2°.

[0063] like Figure 14 As shown, the differential scanning calorimetry spectrum of the eutectic QH-GABA-CCIII-5 has a characteristic endothermic peak at approximately 72.16 °C.

[0064] Example 6 This embodiment illustrates the preparation of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal and coenzyme QH mixture provided by the present invention.

[0065] 186.55 mg of coenzyme QH and 10.77 mg of GABA were added to a mortar, 2 mL of ethanol was added, and the mixture was ground for 2 h. The resulting solid was then dried in a vacuum drying oven at 25 °C for 12 h. The obtained substance was designated as QH-GABA-CCIII-6.

[0066] QH-GABA-CCIII-6 was characterized by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the results are shown in the figures below. Figure 15 and Figure 16 .

[0067] from Figure 15 The X-ray powder diffraction (XRPD) pattern shows that the substance has characteristic peaks at 2θ angles of approximately 2.9°±0.2°, 4.3°±0.2°, 8.6°±0.2°, 10.0°±0.2°, 11.5°±0.2°, 15.7°±0.2°, 17.2°±0.2°, 18.7°±0.2°, 23.3°±0.2°, and 24.1°±0.2°. This indicates that the substance contains reduced coenzyme Q10 / γ-aminobutyric acid cocrystals. It also contains characteristic peaks of coenzyme QH, indicating that the substance is a mixture of reduced coenzyme Q10 / γ-aminobutyric acid cocrystals and coenzyme QH.

[0068] from Figure 16 The differential scanning calorimetry (DSC) spectrum shows that the substance has two characteristic endothermic peaks between 50℃ and 80℃. The endothermic peak at 52.69℃ is consistent with that of coenzyme QH, and the endothermic peak at 72.99℃ is consistent with that of the cocrystal QH-GABA-CCIII mentioned above. This indicates that the substance is composed of reduced coenzyme Q10 / γ-aminobutyric acid cocrystal and coenzyme QH.

[0069] Comparative Example 1 According to the method provided in Example 2 of patent CN113024362A, a reference reduced coenzyme Q10 / nicotinamide cocrystal was prepared. Specifically, 0.4 g of nicotinamide and 1 g of coenzyme QH were added to a mixed solvent of 10 mg isopropanol / isopropyl acetate = 1 / 1 (volume ratio), stirred and dissolved at 40 °C, and recrystallized to obtain a white precipitate. The precipitate was filtered through a Buchner funnel, and the solid was dried in a vacuum drying oven at room temperature for 12 h to obtain the cocrystal of coenzyme QH and nicotinamide, denoted as QH-Nam-CCIII. The cocrystal QH-Nam-CCIII was characterized by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). The results were consistent with those in the appendix of CN113024362A. Figure 1 Appendix Figure 2 Consistent.

[0070] From CN113024362A Figure 2 It is known that the melting point of QH-Nam-CCIII is about 57°C, which is significantly lower than that of the reduced coenzyme Q10 / γ-aminobutyric acid eutectic provided by this invention. Higher melting points often have higher chemical stability, and higher melting points can enable coenzyme QH to withstand higher processing temperatures, thus broadening the application range of coenzyme QH.

[0071] Comparative Example 2 Coenzyme QH and GABA were mixed at a molar ratio of 1:1 at room temperature (25°C) to obtain a coenzyme QH / γ-aminobutyric acid mixture.

[0072] Test Example 1: Short-term stability The cocrystal QH-GABA-CCIII obtained in the above examples, the coenzyme QH obtained in the preparation example, and the coenzyme QH / γ-aminobutyric acid mixture obtained in Comparative Example 2 were placed in an open environment at 25°C and 60% relative humidity to examine their stability. The results are shown in Table 1. The color and shape changes of the cocrystal QH-GABA-CCIII-1 obtained in Example 1 and the coenzyme QH obtained in the preparation example are shown in Table 1. Figure 17 .

[0073] Table 1

[0074] As can be seen from the results in Table 1, the eutectic QH-GABA-CCIII provided by this invention is less prone to oxidation when exposed to air than coenzyme QH, and its stability is significantly better than that of coenzyme QH. Furthermore, the physical mixing of coenzyme QH and GABA alone does not have an antioxidant effect.

[0075] Test Example 2: Long-term / Accelerated Stability The eutectic QH-GABA-CCIII obtained in the above embodiments was sealed in an aluminum-plastic composite bag and placed under conditions of 25℃ / 60%RH and 40℃ / 75%RH to investigate its stability. The results are shown in Table 2.

[0076] Table 2

[0077] As can be seen from Table 2, the eutectic QH-GABA-CCIII provided by this invention exhibits good physical and chemical stability under long-term (25℃ / 60%RH) and accelerated (40℃ / 75%RH) conditions for one month.

[0078] Test Example 3 Particle Size Distribution 1.5g of each of the following cocrystals prepared by the methods in the above examples, coenzyme QH obtained in the preparation example, cocrystal QH-Nam-CCIII obtained in Comparative Example 1, and coenzyme QH / γ-aminobutyric acid mixture obtained in Comparative Example 2 were added to 50mL of ethanol. After thoroughly mixing the samples, they were added to the sample introduction system of a Topsizer Plus laser particle size analyzer (purchased from Zhuhai Omec Instruments Co., Ltd., sample introduction system SCF-126B). The light-blocking level was 15%, and the samples were sonicated for 30s before particle size distribution testing. The results are shown in Table 3. The particle size distribution of coenzyme QH obtained in the preparation example is shown in Table 3. Figure 18 The particle size distribution of the eutectic QH-GABA-CCIII-1 obtained in Example 1 is shown in [reference needed]. Figure 19 .from Figure 18 and Figure 19 It can be seen that the particle size distribution of coenzyme QH is multimodal, while that of the cocrystal QH-GABA-CCIII-1 is unimodal, indicating that the cocrystal QH-GABA-CCIII-1 has a more uniform particle size distribution than coenzyme QH. Particle size distribution typically affects various properties of pharmaceutical formulations, such as dissolution rate, flowability, and bulk density. A uniform particle size distribution is more conducive to controlling drug release rate and improving manufacturability, thereby influencing the safety, efficacy, and quality controllability of the final drug product.

[0079] Table 3

[0080] Test Example 4: Hot Pressing Experiment Weigh 10g each of the eutectic QH-GABA-CCIII obtained from the above examples, the coenzyme QH obtained from the preparation example, the eutectic QH-Nam-CCIII obtained from Comparative Example 1, and the coenzyme QH / γ-aminobutyric acid mixture obtained from Comparative Example 2. Mix each mixture with 90.0g of excipient (microcrystalline cellulose: lactose = 1:1, w / w) in a high-efficiency mixer for 10 minutes to ensure uniformity. Place the mixture in a 60℃ hot air oven for 30 minutes to simulate the thermal environment the material may experience before tableting. After removal, add 0.5% (w / w) magnesium stearate as a lubricant and mix again for 2 minutes. Compress the above mixtures using a single-punch tablet press. Set the punch temperature to 65℃ and the pressure to 10kN. After tableting, hold for about half a minute and observe the tablet formation. The tablet formation states of the eutectic QH-GABA-CCIII-1 obtained in Example 1 and the eutectic QH-Nam-CCIII obtained in Comparative Example 1 are as follows: Figure 20 As shown.

[0081] After tableting, the tablets were collected and weighed, and the material yield (total weight of tablets / total weight of feed) was calculated. The results are shown in Table 4.

[0082] The tablets after compression were ground into powder, and the content of coenzyme QH was determined by HPLC and compared with the initial content. The content retention rate after high-temperature tableting was calculated, and the results are shown in Table 4.

[0083] Table 4

[0084] As shown in Table 4, the QH-GABA-CCIII eutectic prepared by this invention maintains a stable crystal morphology due to its melting point being significantly higher than the processing temperature. This ensures excellent powder flowability and compressibility, preventing adhesion to the punch and mold. Material yield and active ingredient content are almost unaffected, which is more conducive to saving materials and controlling the stability of tablet drug loading. However, under the same conditions, the coenzyme QH, QH-Nam-CCIII eutectic, and coenzyme QH / γ-aminobutyric acid mixture have exceeded their melting points, leading to crystal melting and oiling adhesion with excipients. This not only causes severe material loss and yield reduction but also accelerates the degradation of active ingredients due to localized high temperatures and the molten state, resulting in a significant decrease in content retention.

[0085] Depend on Figure 20 It is evident that the tablets formed by the coenzyme QH-GABA-CCIII-1 cocrystal are stably formed, while the tablets of Comparative Example 1 have softened, deformed, and turned yellow due to oxidation. This indicates that the coenzyme QH-GABA-CCIII cocrystal of the present invention can withstand a higher tableting temperature during the tableting process compared to the cocrystal of Comparative Example 1.

[0086] The above results demonstrate that the cocrystal obtained by this invention has a higher melting point, which directly endows coenzyme Q10 crystals with stronger heat resistance. This makes the cocrystal of this invention applicable to a wider range of solid dosage form processes such as high-temperature mixing, hot melt granulation, and high-temperature tableting. It overcomes the technical bottleneck of existing low-melting-point coenzyme QH in processing and provides a reliable guarantee for its application in tablets, capsules and other heat-processing preparations.

[0087] Test Example 5: In vivo pharmacokinetic experiment (1) Test samples: Sample 1, Sample 2, Sample 3, Sample 4, Sample 5, Reference 1, Reference 2, Reference 3.

[0088] Sample 1: The eutectic QH-GABA-CCIII-1 obtained in Example 1; Sample 2: The eutectic QH-GABA-CCIII-2 obtained in Example 2; Sample 3: The eutectic QH-GABA-CCIII-3 obtained in Example 3; Sample 4: The eutectic QH-GABA-CCIII-4 obtained in Example 4; Sample 5: The eutectic QH-GABA-CCIII-5 obtained in Example 5; Reference sample 1: Coenzyme QH obtained from the preparation example; Reference sample 2: The eutectic QH-Nam-CCIII obtained from Comparative Example 1; Reference sample 3: The coenzyme QH / γ-aminobutyric acid mixture obtained from Comparative Example 2.

[0089] (2) Experimental animals and grouping: SPF - level male Kunming mice, 3 months old and weighing 18 - 22 g, were provided by Dongchuang Experimental Animal Science and Technology Service Department in Kaifu District, Changsha City (Animal Use License Number: SYXK(Xiang)2010 - 0010). Animals were operated in accordance with international experimental animal guidelines to reduce their pain during the experiment. Using a completely randomized design, the mice were randomly divided into groups of 6 each, with a total of 8 groups.

[0090] (3) Experimental conditions: Animal experiments were conducted in a shielded environment. During the experiment, the environmental temperature was 23℃ - 24℃, and the humidity was 50% - 56%. The mice had free access to deionized water and standard feed every day.

[0091] (4) Oral administration and sample collection: Mice fasted for 12 h were randomly divided into groups of 6 each, with a total of 8 groups. Samples 1, 2, 3, 4, 5, reference sample 1, reference sample 2, and reference sample 3 were respectively given to the mice by gavage at a dose of 100 mg / kg (calculated based on coenzyme QH). Blood samples were taken from each group at each time point (1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h, 48 h after administration) into heparin - anticoagulated tubes. After centrifugation and separation, the samples were measured.

[0092] (5) Pharmacokinetic experiments: After mice were orally administered samples 1, 2, 3, 4, 5, reference sample 1, reference sample 2, and reference sample 3 respectively, the obtained plasma samples were processed and then analyzed by HPLC - MS / MS. Fitting analysis was performed based on the blood drug concentration results to calculate the blood drug concentration data. The obtained pharmacokinetic parameter results are shown in Table 5.

[0093] Table 5

[0094] The area under the blood drug concentration - time curve (AUC) is a key indicator for evaluating bioavailability. As can be seen from Table 5, the coenzyme QH - GABA - CCIII cocrystal significantly increased the content of QH in plasma compared with coenzyme QH, QH - Nam - CCIII, and the coenzyme QH / γ - aminobutyric acid mixture, indicating that the coenzyme QH - GABA - CCIII cocrystal has obvious advantages in bioavailability.

[0095] Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and should not be construed as limitations of the present invention. Those of ordinary skill in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention without departing from the principles and purposes of the present invention.

Claims

1. A reduced coenzyme Q10 / γ-aminobutyric acid cocrystal, characterized in that, The reduced coenzyme Q10 / γ-aminobutyric acid cocrystal is a cocrystal formed by reduced coenzyme Q10 and γ-aminobutyric acid.

2. The reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to claim 1, characterized in that, The X-ray powder diffraction pattern of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal obtained by Cu-Kα radiation has characteristic peaks at at least the following 2θ angles: 2.9°, 4.3°, 8.6°, 10.0°, 11.5°, 15.7°, 17.2°, 18.7°, 23.3°, and 24.1°, with an error of ±0.2° for the 2θ angle.

3. The reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to claim 1, characterized in that, The differential scanning calorimetry spectrum of the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal has a characteristic endothermic peak at least at 73℃±2℃.

4. The reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to claim 1, characterized in that, The reduced coenzyme Q10 / γ-aminobutyric acid cocrystal exhibits a unimodal particle size distribution, ranging from 0.01 μm to 20 μm.

5. The reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to any one of claims 1-4, characterized in that, The stoichiometric ratio of reduced coenzyme Q10 to γ-aminobutyric acid in the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal is 1:(0.5-2.5).

6. The method for preparing the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to any one of claims 1-5, characterized in that, The method includes recrystallizing reduced coenzyme Q10 and γ-aminobutyric acid together in a solvent to obtain a reduced coenzyme Q10 / γ-aminobutyric acid cocrystal, or grinding reduced coenzyme Q10 and γ-aminobutyric acid to obtain a reduced coenzyme Q10 / γ-aminobutyric acid cocrystal.

7. The method for preparing reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to claim 6, characterized in that, The molar ratio of reduced coenzyme Q10 to γ-aminobutyric acid is 1:(0.5-2.5).

8. The method for preparing reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to claim 6, characterized in that, The solvent is selected from at least one of water, alcohol solvents, nitrile solvents, ketone solvents, ester solvents, alkane solvents, aromatic hydrocarbon solvents, and haloalkane solvents.

9. The method for preparing reduced coenzyme Q10 / γ-aminobutyric acid cocrystal according to claim 6, characterized in that, The recrystallization method includes dissolving reduced coenzyme Q10 and γ-aminobutyric acid together in a solvent, then stirring and cooling to crystallize or allowing it to stand to evaporate and crystallize. Preferably, the conditions for stirring and dissolving include a temperature of 20℃-80℃ and a time of 0.1h-2h; Preferably, the conditions for stirring, cooling, and crystallization include a temperature of 4℃-40℃ and a time of 1h-48h. Preferably, the conditions for static evaporation and crystallization include a temperature of 4℃-40℃ and a time of 1 day-30 days; Preferably, the grinding is dry grinding or wet grinding.

10. A coenzyme Q10 product, characterized in that, The coenzyme Q10 product contains the reduced coenzyme Q10 / γ-aminobutyric acid cocrystal as described in any one of claims 1-5.

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