Cardioplegic solution composition containing coenzyme q 10 and use thereof

Abstract

A cardioplegic solution composition containing coenzyme Q10, and a use thereof. the cardioplegic solution composition containing coenzyme Q10 comprising a coenzyme Q10 solution and a cardioplegic solution. In the coenzyme Q10 solution, the solvent comprises 33-60 parts by weight of a pharmaceutically acceptable polyoxyethylene nonionic surfactant and 40-67 parts by weight of a water-soluble polyhydroxy compound; the concentration of coenzyme Q10 is 14.8-58.8 μg / mg, and the obtained solution is a clear solution having an HLB value of 9-29. The cardioplegic solution composition containing coenzyme Q10 is used for the preservation of an ex vivo donor heart in heart transplantation, and and is also used in a cardioplegic solution drug for perfusion during establishment of extracorporeal circulation.

Description

A coenzyme Q 10 Cardioplegic solution composition and application thereof

[0001] This application claims the benefit of Chinese patent application No. 2023115435077, filed on November 20, 2023. This application incorporates the entirety of the aforementioned Chinese patent application. Technical Field

[0002] The present invention relates to the field of biomedicine technology, and in particular to a method for preparing a biopharmaceutical product containing coenzyme Q 10 Cardioplegic solution composition and application thereof. Background Art

[0003] Coenzyme Q 10 It is a fat-soluble quinone compound with common vitamin characteristics and a chemical structure similar to vitamin K. It has multiple biological functions, especially its advantages of small dosage, low toxicity, and auxiliary treatment of various diseases, and has been widely used in clinical practice. 10 It is a good biochemical drug with natural antioxidant and cell metabolism activation effects, which can significantly improve human immunity. This drug is mainly used clinically for the treatment of cardiovascular diseases, scurvy, aplastic anemia, duodenal ulcers, acute and chronic viral hepatitis, subacute liver necrosis, congestive heart disease, emphysema and other diseases, as well as auxiliary treatment for cancer patients. It is widely used in nutritional health products and food additives at home and abroad.

[0004] But Coenzyme Q 10 In terms of solubility, it has the physical and chemical characteristics of being very soluble in chloroform, benzene, easily soluble in acetone, ether, difficult to dissolve in ethanol, insoluble in water and methanol, and due to the coenzyme Q 10 There is a quinone group in the molecular structure of coenzyme Q, which is easily decomposed by light and oxygen. 10 Solubility requires more excipients, such as surfactants, organic solvents or long-chain fatty acids (oils) or lipids. 10 Intravenous infusion and preparation method thereof, which provides a method for infusing coenzyme Q 10 The method of making intravenous infusion is mainly to add high concentration of surfactant polysorbate 80 (Tween 80) to coenzyme Q 10 Polysorbate 80 (Tween 80) is a commonly used coenzyme Q 10 Solubilizing excipient, used alone to dissolve coenzyme Q 10 , can keep the solution clear in a short time, but after a few days, coenzyme Q 10 Will be reprecipitated. 10Intravenous Infusion Solution”, which provides another way to 10 The key to the method of preparing intravenous infusion is the use of a composite solubilizer, which is composed of polysorbate 80 (Tween 80) and polyoxyethylene fatty acid ester (polyoxyethylene fatty acid 40 ester) in a certain proportion. 10 The injection uses a solubilizing emulsifier and a surfactant polysorbate to increase the content of coenzyme Q 10 Patent CN200810069760.2 Coenzyme Q 10 The injection utilizes a mixture of polyethylene glycol 15-hydroxystearate (Solutol HS 15) and polysorbate 80. These patented technologies all utilize a combination of multiple solubilizing agents, and these ingredients are used in relatively large quantities, raising concerns about their safety and impact on cardiomyocytes. Furthermore, excessive amounts of polysorbate 80 can lead to safety concerns, such as allergic reactions.

[0005] Another example is the use of micelle solubilization, such as coenzyme Q 10 Encapsulated in a micelle containing glycyrrhizic acid or glycyrrhizic acid salt, bile acid and unsaturated salt (Coenzyme Q 10 Solubilizing composition and preparation method thereof, CN201780024843.7), or adding vegetable oil to prepare an emulsion. Some researchers also used coenzyme Q 10 Mix with polyethylene glycol dodecyl hydroxystearate solubilizer, heat until melted, then add injection solvent to make a clear solution (a coenzyme Q 10 Intravenous infusion and its preparation method, CN101480375A).

[0006] However, the cardioplegic solution has a complex formula, containing 0.8766g sodium chloride, 0.6710g potassium chloride, 0.1842g 2-oxoglutarate-hydrogen-potassium, 0.8132g magnesium chloride hexahydrate, 3.7733g histidine hydrochloride monohydrate, 27.9289g histidine, 0.4085g tryptophan, 5.4651g mannitol, and 0.0022g calcium chloride dihydrate per 1000mL solution. Therefore, unlike the intravenous injection solution described in the above patent (usually using 0.9% sodium chloride solution or 5% glucose solution), after the coenzyme Q10 solubilized preparation is added to the cardioplegic solution, it may aggregate due to interaction with ions or molecules in the cardioplegic solution, resulting in the encapsulated coenzyme Q 10 At the same time, the types and amounts of excipients used in these preparations cannot meet the safety requirements for cardioprotection.

[0007] The complexity of the cardioplegic solution ingredients also requires that when improving its formula, it is necessary to ensure that the content of the original ingredients remains unchanged while minimizing the types and contents of the newly added ingredients to reduce the disturbance of the ionic components in the cardioplegic solution and thus affect the cardioplegic effect of the cardioplegic solution. Therefore, the present invention gives priority to using a single and small amount of solubilizer as the main material to improve the coenzyme Q 10 Solubility in water, dissolving coenzyme Q 10 While reducing the impact on the components of the cardioplegic solution itself. Taking into account the types and contents of ions in the cardioplegic solution and the safety requirements of the cardioplegic solution for cardiac protection, the present invention selects a polyoxyethylene nonionic surfactant with good safety as a solubilizer to reduce the impact of ions on the stability of the coenzyme Q10 solubilized preparation; further, a water-soluble polyhydroxy compound is added as a cosolvent, utilizing the hydrogen bonding between the polyhydroxy compound and the hydrophilic groups of the polyoxyethylene surfactant (Figure 1), while reducing the amount of surfactant used, ensuring that coenzyme Q10 can be solubilized in the cardioplegic solution for a long time.

[0008] In terms of stability, due to the coenzyme Q 10 It is very easy to be oxidized. The existing technology mainly maintains the coenzyme Q by adding sodium bisulfite and sodium calcium edetate as reducing agents. 10 HTK cardioplegic solution primarily achieves cardiac arrest by inhibiting the action potential of myocardial cells through high concentrations of potassium ions and low concentrations of sodium and calcium ions in the solution. Therefore, directly adding these two ionic reducing agents to the cardioplegic solution will alter the ion concentration within the solution, affecting its effectiveness. If the sodium ion concentration in the cardioplegic solution increases, the tendency for extracellular sodium ions to diffuse into the cell increases energy consumption by the sodium-potassium pump. The reduced ATP further increases the risk of cellular edema during cardioplegic arrest. Summary of the Invention

[0009] The purpose of the present invention is to provide a coenzyme Q 10 The cardioplegic solution composition has high drug loading and stability.

[0010] Another object of the present invention is to provide applications of the above composition.

[0011] The object of the present invention is achieved through the following scheme: The present invention provides a coenzyme Q 10 A solution, wherein the dissolving agent comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant and a water-soluble polyhydroxy compound;

[0012] The polyoxyethylene nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic / capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride;

[0013] The water-soluble polyol comprises one or more of glycerol, polyethylene glycol, polyglycerol, copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol. 10 In the solution, the dissolving agent comprises 33 to 60 parts by weight of a pharmaceutically acceptable polyoxyethylene nonionic surfactant and 40 to 67 parts by weight of a water-soluble polyhydroxy compound.

[0014] In some embodiments, the coenzyme Q 10 The solution comprises 40 parts, 60 parts, 33.3 parts or 50 parts by weight of a polyoxyethylene nonionic surfactant as a solubilizer.

[0015] In some embodiments, the coenzyme Q 10 In the coenzyme Q 10 The concentration in the solution is 14.8 to 58.8 μg / mg, for example, 19.6 μg / mg, 38.5 μg / mg, 24.3 μg / mg, 14.8 μg / mg, 58.8 μg / mg or 24.4 μg / mg.

[0016] In some embodiments, the coenzyme Q 10 The raw materials of the solution are any of the following formulas:

[0017] Formula a. Polyoxyethylene hydrogenated castor oil, glycerin and coenzyme Q 10 ;

[0018] Formulation b. Polyoxyethylated 12-hydroxystearic acid, polyethylene glycol 400 and coenzyme Q 10 ;

[0019] Formula c. Oleoyl polyoxyethylene glyceride, glycerol and coenzyme Q 10 ;

[0020] Formula d. Lauroyl polyoxyethylene-6 glyceride, glycerin and coenzyme Q 10 ;

[0021] Formula e. Polyoxyethylene castor oil, glycerin and coenzyme Q 10 ;

[0022] Formula f. Caprylic / capric macrogol glycerides, polyethylene glycol 300 and coenzyme Q 10 .

[0023] In some embodiments, the coenzyme Q 10 The raw materials of the solution are any of the following formulas:

[0024] Formula 1. 400mg polyoxyethylene hydrogenated castor oil, 600mg glycerin and 20mg coenzyme Q 10 ;

[0025] Formulation 2. 600 mg polyoxyethylated 12-hydroxystearic acid, 400 mg polyethylene glycol 400, and 40 mg coenzyme Q 10 ;

[0026] Formula 3: 200mg oleoyl polyoxyethylene glycerides, 400mg glycerol, and 15mg coenzyme Q 10 ;

[0027] Formula 4. 400mg lauroyl polyoxyethylene-6 glyceride, 600mg glycerin and 15mg coenzyme Q 10 ;

[0028] Formula 5. 400mg polyoxyethylene castor oil, 400mg glycerin, 50mg coenzyme Q 10 ;

[0029] Formula 6: 200mg caprylic / capric macrogol glycerides, 400mg polyethylene glycol 300, and 15mg coenzyme Q 10 .

[0030] The present invention also provides a coenzyme Q 10 Cardioplegic solution composition comprising coenzyme Q 10 Solutions and cardioplegic solutions, wherein

[0031] The coenzyme Q 10 A solution, wherein the dissolving agent comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant and a water-soluble polyhydroxy compound, and the concentration of the dissolving agent in the cardioplegia solution is no greater than 10 mg / mL;

[0032] The polyoxyethylene nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic / capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride;

[0033] The water-soluble polyol includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

[0034] In some embodiments, the coenzyme Q 10 Cardioplegic solution composition comprising coenzyme Q 10 Solutions and cardioplegic solutions, wherein

[0035] The coenzyme Q 10 A solution, wherein the dissolving agent comprises 33 to 60 parts by weight of a pharmaceutically acceptable polyoxyethylene nonionic surfactant and 40 to 67 parts by weight of a water-soluble polyhydroxy compound, wherein the concentration of the dissolving agent in the cardioplegia solution is no greater than 10 mg / mL;

[0036] The polyoxyethylene nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic / capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride;

[0037] The water-soluble polyol includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

[0038] In some embodiments, the coenzyme Q 10 Cardioplegic solution composition comprising coenzyme Q 10 Solutions and cardioplegic solutions, wherein

[0039] The coenzyme Q 10 The solution comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant in parts by weight of 33 to 60 parts and a water-soluble polyhydroxy compound in parts by weight, coenzyme Q 10 The concentration is 14.8 to 58.8 μg / mg, the concentration of the dissolving agent in the cardioplegia solution is not greater than 10 mg / mL, and the coenzyme Q 10 The concentration in the cardioplegic solution is 15 to 60 mg / L;

[0040] The polyoxyethylene nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic / capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride;

[0041] The water-soluble polyol includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

[0042] In some embodiments, the coenzyme Q 10 Cardioplegic solution composition comprising coenzyme Q 10 Solutions and cardioplegic solutions, wherein

[0043] The coenzyme Q 10 The solution comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant in parts by weight of 33 to 60 parts and a water-soluble polyhydroxy compound in parts by weight, coenzyme Q 10 The concentration is 14.8 to 58.8 μg / mg, and the resulting solution is a clear solution with an HLB value of 9 to 29. The concentration of the dissolving agent in the cardioplegia solution is not greater than 10 mg / mL, and coenzyme Q 10 The concentration in the cardioplegic solution is 15 to 60 mg / L;

[0044] The polyoxyethylene nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic / capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride;

[0045] The water-soluble polyol includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

[0046] In some embodiments, the coenzyme Q 10 Cardioplegic solution composition comprising coenzyme Q 10 Solutions and cardioplegic solutions, wherein

[0047] The coenzyme Q 10 The solution comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant in parts by weight of 33 to 60 parts and a water-soluble polyhydroxy compound in parts by weight, coenzyme Q10 The concentration is 14.8-58.8 μg / mg, and the resulting solution is a clear solution with an HLB value of 9-29. 10 The self-assembly with the surfactant forms micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure; the concentration of the dissolving agent in the cardioplegia solution is not more than 10 mg / mL, and the coenzyme Q 10 The concentration in the cardioplegia solution is 15 to 60 mg / L, for example, the coenzyme Q 10 The concentrations are 15mg / L, 17mg / L, 20mg / L, 23mg / L, 25mg / L, 27mg / L, 30mg / L, 35mg / L, 40mg / L, 45mg / L, 50mg / L, 55mg / L and 60mg / L;

[0048] The polyoxyethylene nonionic surfactant as a solubilizer includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic / capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride;

[0049] The water-soluble polyol includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

[0050] In some embodiments, the coenzyme Q 10 Cardioplegic solution composition comprising coenzyme Q 10 solutions and cardioplegic solutions,

[0051] The coenzyme Q 10 The solution comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant as a solubilizer in 33 to 60 parts by weight and a water-soluble polyhydroxy compound as a cosolvent in 40 to 67 parts by weight, and coenzyme Q 10 The concentration is 14.8~58.8μg / mg, and the resulting solution is a clear solution with an HLB value of 9~29. 10 The self-assembly with the surfactant forms micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure; the concentration of the dissolving agent in the cardioplegia solution is not more than 10 mg / mL, and the coenzyme Q 10 The concentration is 15-60 mg / L;

[0052] The polyoxyethylene nonionic surfactant as a solubilizer includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic / capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride;

[0053] The water-soluble polyhydroxy compound as a cosolvent includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

[0054] In some embodiments, the coenzyme Q 10 The solution comprises a dissolving agent comprising 40 parts, 60 parts, 33.3 parts or 50 parts by weight of a polyoxyethylene nonionic surfactant as a solubilizing agent.

[0055] In some embodiments, the coenzyme Q 10 In the coenzyme Q 10 The concentration in the solution is 14.8 to 58.8 μg / mg, for example, 19.6 μg / mg, 38.5 μg / mg, 24.3 μg / mg, 14.8 μg / mg, 58.8 μg / mg or 24.4 μg / mg.

[0056] Coenzyme Q obtained 10 The solution is added to water or aqueous solution, through coenzyme Q 10 It self-assembles with surfactants to form micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure. Therefore, only a small amount of surfactant is needed to greatly increase the coenzyme Q in the micelle-like structure. 10 It is helpful to reduce the amount of excipients in the solution and increase the coenzyme Q 10 Stability of the solution.

[0057] Furthermore, the solubilizer and cosolvent in the dissolving agent are mixed with coenzyme Q at 40-50 ° C. 10 Mix well to obtain coenzyme Q 10 Solution, the resulting coenzyme Q 10 The solution exists in a semi-solid form at low temperature. After being heated in a water bath, it returns to a clear solution and is added to the cardiac protective solution. It self-assembles with the surfactant to form a solution with coenzyme Q wrapped inside. 10 The micelle-like self-assembly structure makes coenzyme Q 10 Stably dispersed in the cardioprotective solution, this coenzyme Q 10 Solution of Coenzyme Q 10 Problems of being insoluble in water and easily oxidized.

[0058] Coenzyme Q 10 The solubilizer consists of two parts: a solubilizer and a cosolvent. The solubilizer is a non-ionic surfactant with a polyethylene glycol structure as the hydrophilic group and an HLB value between 9 and 29; the cosolvent is a polyhydroxy hydrophilic component, such as polyethylene glycol 300 and 400, and glycerol (propylene glycol).

[0059] Coenzyme Q 10 The solubilizer comprises a solubilizer and a cosolvent. The solubilizer is a nonionic surfactant with a polyethylene glycol structure as the hydrophilic group and an HLB value between 9 and 29, such as 9, 14, 16, 13.5, or 12. The cosolvent is a polyhydroxy hydrophilic component, such as polyethylene glycol 300 and 400, and glycerol (propylene glycol).

[0060] Based on the above solution, the weight average molecular weight of the polyethylene glycol is 200-1000.

[0061] The present invention gives priority to using a single and small amount of solubilizing agent as the main material to improve the coenzyme Q 10 Solubility in water, can be dissolved in coenzyme Q 10 While reducing the impact on the components of the cardioplegic solution itself.

[0062] Preferably, the concentration of the dissolving agent in the cardioplegic solution is 1.1 mg / mL to 10 mg / mL.

[0063] Preferably, the coenzyme Q 10 The concentration is 15-30 mg / L.

[0064] In some embodiments, the coenzyme Q 10 The raw materials of the solution are any of the following formulas:

[0065] Formula a. Polyoxyethylene hydrogenated castor oil, glycerin and coenzyme Q 10 ;

[0066] Formulation b. Polyoxyethylated 12-hydroxystearic acid, polyethylene glycol 400 and coenzyme Q 10 ;

[0067] Formula c. Oleoyl polyoxyethylene glyceride, glycerol and coenzyme Q 10 ;

[0068] Formula d. Lauroyl polyoxyethylene-6 glyceride, glycerin and coenzyme Q 10 ;

[0069] Formula e. Polyoxyethylene castor oil, glycerin and coenzyme Q 10 ;

[0070] Formula f. Caprylic / capric macrogol glycerides, polyethylene glycol 300 and coenzyme Q 10 .

[0071] In some embodiments, the coenzyme Q 10 The raw materials of the solution are any of the following formulas:

[0072] Formula 1. 400mg polyoxyethylene hydrogenated castor oil, 600mg glycerin and 20mg coenzyme Q 10 ;

[0073] Formulation 2. 600 mg polyoxyethylated 12-hydroxystearic acid, 400 mg polyethylene glycol 400, and 40 mg coenzyme Q 10 ;

[0074] Formula 3: 200mg oleoyl polyoxyethylene glycerides, 400mg glycerol, and 15mg coenzyme Q 10 ;

[0075] Formula 4. 400mg lauroyl polyoxyethylene-6 glyceride, 600mg glycerin and 15mg coenzyme Q 10 ;

[0076] Formula 5. 400mg polyoxyethylene castor oil, 400mg glycerin, 50mg coenzyme Q 10 ;

[0077] Formula 6: 200mg caprylic / capric macrogol glycerides, 400mg polyethylene glycol 300, and 15mg coenzyme Q 10 .

[0078] Furthermore, the composition also contains thiourea to further increase the coenzyme Q 10 Stability in the composition.

[0079] Preferably, the concentration of thiourea is 0.06 mg / mL to 0.54 mg / mL.

[0080] Furthermore, the cardioplegic solution is an intracellular solution, such as HTK cardioplegic solution. 10 The solution can be well dissolved in HTK stop solution without aggregation and precipitation.

[0081] The present invention uses non-ionic reducing agent thiourea to maintain coenzyme Q without affecting the cardiac arrest effect of the cardioplegia solution. 10 The coenzyme Q 10 Thiourea can further stabilize it in HTK liquid and prevent it from oxidative deterioration.

[0082] In some embodiments, the coenzyme Q 10The cardioplegia solution composition can be prepared by the following method, which comprises the following steps: 10 The solution is mixed with the cardioplegia solution to obtain the coenzyme Q 10 The cardioplegic solution composition can be used.

[0083] In some embodiments, the coenzyme Q 10 In the solution, the dissolving agent comprises 33 to 60 parts by weight of a pharmaceutically acceptable polyoxyethylene nonionic surfactant and 40 to 67 parts by weight of a water-soluble polyhydroxy compound.

[0084] In some embodiments, the coenzyme Q 10 The solution can be obtained by the following method: the polyoxyethylene nonionic surfactant, the water-soluble polyhydroxy compound and coenzyme Q 10 Mix and heat to obtain the coenzyme Q 10 solution.

[0085] In some embodiments, the heating temperature is 30-60°C, preferably 40°C or 50°C, such as 50°C.

[0086] The present invention also provides a coenzyme Q-containing 10 The preparation method of the cardioplegia solution composition comprises the following steps: 10 The solution is mixed with the cardioplegia solution to obtain the coenzyme Q 10 The cardioplegic solution composition can be;

[0087] The operation and conditions of the preparation method are as described in any of the above schemes.

[0088] The present invention also provides application of the composition in preservation of isolated donor hearts in heart transplantation.

[0089] The present invention also provides the use of the composition in the preparation of a cardioplegic solution for perfusion during extracorporeal circulation. 10 Cardiac arrest solution has a protective effect on myocardial cells after ischemia-reperfusion.

[0090] The mechanism of the present invention is: using polyoxyethylene surfactant as a solubilizer, solubilizing through micelles, and improving the coenzyme Q 10 Solubility in aqueous solution, where the hydrophilic structure of polyoxyethylene surfactants is formed by polymerization of ethylene glycol; polyhydroxy polymers, such as glycerol, polyethylene glycol, etc., have hydroxyl groups that can form hydrogen bonds with polyoxyethylene groups (Formula 1). Polyhydroxy polymers stabilize coenzyme Q through hydrogen bonding. 10 The micelle structure allows the use of less surfactant to bind coenzyme Q 10Stably solubilized in micelles.

[0091] Formula 1 is the coenzyme Q of the present invention 10 Reaction mechanism of the solution:

[0092] In Formula 1, a polyhydroxy polymer (using glycerol as an example) forms intermolecular hydrogen bonds with a polyoxyethylene surfactant (e.g., polyoxyethylene hydrogenated castor oil containing polyoxyethylene groups (Cremophor RH40) or Pluronic F68). The dotted box represents the hydrophilic structure of this type of surfactant.

[0093] The advantages of the present invention are: solving the problem of coenzyme Q 10 Solubility, stability and efficacy in cardioplegic solution.

[0094] (1) The present invention solves the problem of coenzyme Q 10 Solubility issues in cardioplegic solutions:

[0095] By preparing coenzyme Q with high drug loading 10 solution, exists in semi-solid form at low temperature; the coenzyme Q 10 After the solution is heated in a water bath, it returns to a clear solution and is then added to the cardiac protective solution or cardioplegia solution according to the required concentration. 10 Self-assembled with surfactant to form a coenzyme Q encapsulated inside 10 micelle-like self-assembly structure, thereby making coenzyme Q 10 Can be stably dispersed in the heart protection solution. 10 The solution solves the problem that coenzyme Q10 is insoluble in water and easily oxidized.

[0096] (2) The present invention improves coenzyme Q 10 Compatibility in cardioplegic solutions:

[0097] The present invention uses coenzyme Q 10 The drug and / or its analogs self-assemble with surfactants to form micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure. Therefore, only a small amount of surfactant is needed to greatly increase the coenzyme Q in the micelle-like structure. 10 This is beneficial for reducing the amount of excipients in the solution, thereby improving the compatibility of the solution with cardiac protective solutions or cardioplegic solutions when used for myocardial protection.

[0098] (3) The present invention improves the stability of drug loading:

[0099] The present invention is used to carry out coenzyme Q 10 During loading, the micelle-like structure dispersed in water has a restrictive effect on particle growth, making the coenzyme Q dissolved in the cosolvent10 Before the drug can aggregate into large particles, it is effectively loaded into the micelle-like structure in the form of molecules or microcrystals. The drug in the microcrystal state not only greatly increases the drug loading capacity, but also further improves the stability of the drug loading, which can avoid the 10 Precipitation in cardiac protective solution or cardioplegia solution. In addition, the above-mentioned coenzyme Q provided by the present invention 10 The solution avoids the use of large amounts of oil and lipid components and has good stability and safety. 10 The solution preparation process is simple and easy to expand production. BRIEF DESCRIPTION OF THE DRAWINGS

[0100] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0101] Figure 1 is the coenzyme Q provided in Example 1 10 Dissolving agents (A and B, respectively, formulations 1 and 2 in Example 1) and solutions diluted with cardioplegic solution (C, 1, 2, 3, 7, respectively corresponding to the numbers of the formulations in Example 1);

[0102] Figure 2 shows the coenzyme Q prepared by several groups of formulas provided in Example 1 10 Particle size distribution of particles in the solution obtained by diluting the dissolving agent with the cardioplegic solution; A to C are samples prepared in formulations 1, 2, and 7 in Example 1, respectively;

[0103] Figure 3, A to E, shows flow cytometry analysis of human induced pluripotent stem cell-derived cardiomyocytes. A shows the selection of live cell population P1 based on forward scattered light and side scattered light, while dead cells are removed. B shows the selection of diagonal cell population P2 based on forward scattered light height and area, while adherent cell populations are removed. C shows the gated control cell population unlabeled with cardiac troponin T antibody. D and E show the identification of cardiac troponin T-positive cardiomyocyte population P3 based on the gate in C. Flow cytometry showed that cardiac troponin T-positive cardiomyocytes accounted for 85.9%. F shows the action potential of human induced pluripotent stem cell-derived cardiomyocytes.

[0104] Figure 4 , A is the Western blot analysis results of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) after cardiomyocytes were cultured in HTK cardioplegia solution containing 10 mg / mL, 3.3 mg / mL, and 1.1 mg / mL of lytic agent; B is a bar graph of Western blot analysis of apoptosis pathway activation levels; C is a TUNEL staining image of cardiomyocytes, where C1-C4, C5-C8, C9-C12, and C13-C16 are TUNEL staining images of cardiomyocytes cultured in HTK cardioplegia solution containing 10 mg / mL, 3.3 mg / mL, and 1.1 mg / mL of lytic agent, respectively, with DAPI (nuclei), TUNEL (apoptotic cells), Actinin (cytoskeleton), and Merge (overlay of DAPI, TUNEL, and Actinin staining), with scale bar 50 μm; D is a bar graph of cardiomyocyte apoptosis rate detected by TUNEL staining;

[0105] In Figure 5, A is the Western blot detection results of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) after myocardial cells were cultured in HTK cardioplegia solution containing 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL thiourea; B is the Western blot detection results of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) after myocardial cells were cultured in HTK cardioplegia solution containing 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL thiourea; Blot analysis of the activation level of the apoptosis pathway; C is a TUNEL staining image of cardiomyocytes, where C1-C4, C5-C8, C9-C12, and C13-C16 are TUNEL staining images of cardiomyocytes cultured in HTK cardioplegia solution containing 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL thiourea, respectively. DAPI (nucleus), TUNEL (apoptotic cells), Actinin (cytoskeleton), and Merge (overlay of DAPI, TUNEL, and Actinin staining) are shown. Scale bar: 50 μm; D is a bar graph of the apoptosis rate of cardiomyocytes detected by TUNEL staining.

[0106] Figure 6: A, B, C, and D are coenzyme Q in HTK cardioplegia solutions containing 0.54 mg / mL, 0.18 mg / mL, 0.06 mg / mL thiourea, and no thiourea, respectively. 10 E, F, and G are the curves of changes in the contents of α-ketoglutarate, tryptophan, and histidine in HTK cardioplegia solution containing 0.54 mg / mL thiourea over time;

[0107] Figure 7: A, B, C, and D are respectively myocardial cells perfused with HTK cardioplegia solution (A), HTK cardioplegia solution containing 0.54 mg / mL thiourea (B), and HTK cardioplegia solution containing 60 mg / L coenzyme Q 10 Action potential after HTK cardioplegia (C) and HTK cardioplegia containing 2.5 mg / mL sodium bisulfite (D). The arrow pointing to the lower left indicates that the myocardial cell begins to stop electrophysiological activity, and the arrow pointing to the lower right indicates that the myocardial cell restarts action potential.

[0108] Figure 8: A shows cardiomyocytes in HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L, and 60 mg / L coenzyme Q 10 Western blot detection results of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) in the HTK cardioplegia solution composition after ischemia-reperfusion; B is a bar graph of the activation level of the apoptosis pathway detected by Western blot; C is a TUNEL staining diagram of myocardial cells, where C1-C4, C5-C8, C9-C12, C13-C16, and C17-C20 are the activation levels of myocardial cells in the HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L, and 60 mg / L coenzyme Q 10 Figure 3 TUNEL staining of HTK cardioplegia solution after ischemia-reperfusion, DAPI (nucleus), TUNEL (apoptotic cells), Actinin (cytoskeleton), and Merge (overlay of DAPI, TUNEL, and Actinin staining), scale bar, 50 μm; D is a bar graph of TUNEL staining to detect myocardial cell apoptosis rate; *P < 0.05, vs HTK;

[0109] Figure 9: A shows cardiomyocytes in HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L, and 60 mg / L coenzyme Q 10 Western blot detection results of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) in the HTK cardioplegia solution composition after ischemia-reperfusion; B is a bar graph of the activation level of the apoptosis pathway detected by Western blot; C is a TUNEL staining diagram of myocardial cells, where C1-C4, C5-C8, C9-C12, C13-C16, and C17-C20 are the activation levels of myocardial cells in the HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L, and 60 mg / L coenzyme Q 10Figure 4 TUNEL staining of HTK cardioplegia solution after ischemia-reperfusion, DAPI (nucleus), TUNEL (apoptotic cells), Actinin (cytoskeleton) and Merge (overlay of DAPI, TUNEL and Actinin staining), scale bar, 50 μm; D is a bar graph of the apoptosis rate of myocardial cells detected by TUNEL staining; *P < 0.05, vs HTK. DETAILED DESCRIPTION

[0110] Each of the following examples is intended to illustrate, not to limit, the present invention. Indeed, it will be apparent to those skilled in the art that various modifications and variations may be made to the present invention without departing from the scope or spirit of the invention. For example, features illustrated or described as part of one embodiment may be used in another embodiment to produce a further embodiment.

[0111] Unless otherwise indicated, all terms (including technical and scientific terms) used to disclose the present invention have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. By way of further guidance, the following definitions are provided to better understand the teachings of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0112] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the arbitrary and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical and" and also undoubtedly includes technical solutions connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution of all being connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution of all being connected by "logical AND").

[0113] As used herein, the terms "comprising," "including," and "comprising" are synonymous and are inclusive or open-ended and do not exclude additional, unrecited members, elements, or method steps.

[0114] The recitation of numerical ranges herein by endpoints includes all numbers and fractions subsumed within the range, as well as the recited endpoints.

[0115] The present invention involves concentration values, and its meaning includes fluctuations within a certain range. For example, it can fluctuate within the corresponding accuracy range. For example, 2% can allow fluctuations within the range of ±0.1%. For values ​​that are large or do not require overly precise control, its meaning is also allowed to include larger fluctuations. For example, 100mM can allow fluctuations within the range of ±1%, ±2%, ±5%, etc. As for molecular weight, its meaning is allowed to include fluctuations of ±10%. In the present invention, the word "about" is used as a modifier of quantity to indicate that it includes + or -5% of the modified quantity.

[0116] In the present invention, descriptions such as "plurality" and "multiple" refer to a number greater than or equal to 2 unless otherwise specified.

[0117] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0118] In the present invention, "preferably", "better", "more preferably", and "suitably" are merely descriptions of preferred implementation methods or examples, and should be understood to not limit the scope of protection of the present invention. In the present invention, "optionally", "optional", and "optional" refer to being optional, that is, to being selected from either of the two parallel options of "with" or "without". If multiple "options" appear in a technical solution, unless otherwise specified and without contradiction or mutual restriction, each "optional" is independent.

[0119] The present invention relates to a coenzyme Q 10 The solution comprises a solvent comprising 33 to 60 parts by weight of a pharmaceutically acceptable polyoxyethylene nonionic surfactant and 40 to 67 parts by weight of a water-soluble polyhydroxy compound.

[0120] The solution is added to water or aqueous solution to 10 It self-assembles with surfactants to form micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure. Therefore, only a small amount of surfactant is needed to greatly increase the coenzyme Q in the micelle-like structure. 10 This is beneficial to reduce the amount of excipients in the solution, so as to improve the compatibility of the solution with the heart when used for myocardial protection. 10 During loading, the micelle-like structure dispersed in water also has a limiting effect on particle growth, making the coenzyme Q dissolved in the solvent 10Before the drug can aggregate into large particles, it is effectively loaded into the micelle-like structure in the form of molecules or microcrystals. The drug in the microcrystal state not only greatly increases the drug loading capacity, but also further improves the stability of the drug loading, which can avoid the 10 In addition, the above-mentioned coenzyme Q provided by the present invention 10 The solution avoids the use of a large amount of oil and lipid components, has good stability and safety. It also has the advantages of simple process and easy scale-up. 10 " should be understood to include pharmaceutical analogs or derivatives thereof.

[0121] As used herein, the terms "micelle-like self-assembled structure," "micelle-like particle," "micelle-like structure," "micelle-like drug," or "self-assembled structure" refer to a dispersed system formed by the spontaneous, oriented arrangement of surfactants, optionally in combination with drug molecules, at an interface in a medium. The surface activity of surfactants stems from the amphiphilic structure of their molecules. The hydrophilic groups induce the molecules to enter the aqueous phase, while the hydrophobic groups prevent them from dissolving in water and instead migrate outward from the interior of the water, tending to escape the aqueous phase. The balance between these two tendencies results in the accumulation of surfactants at the interface. When surface adsorption reaches saturation, the surfactant molecules, under the action of the hydrophobic groups, self-assemble within the solution. Specifically, the hydrophobic groups form an inner core, while the hydrophilic groups face outward in contact with the water, forming the simplest micelle-like self-assembled structure.

[0122] When amphiphilic molecules (including surfactants) encounter a selective solvent (i.e., one that can only dissolve the hydrophilic or hydrophobic segments but not the other), micelles may be formed. There are many kinds of selective solvents that can be used, including, for example, water. In the present invention, the terms "loaded in a micelle-like self-assembled structure" and "self-assembled with a surfactant to form a micelle-like self-assembled structure" are used interchangeably to refer to coenzyme Q. 10 It interacts with surfactants, where the drug molecules are encapsulated in the particles formed by the two, forming a stable drug-loaded micelle-like structure.

[0123] The polyoxyethylene nonionic surfactant suitable for the present invention refers to a surfactant mainly composed of ethylene oxide (EO) and a hydrophobic compound containing an active hydrogen atom, and combined into a surfactant of any length as needed, which mainly plays a solubilizing role. Its content can be 33 to 60 parts, for example 40, 45, 50, or 55 parts. Surfactants suitable for the present invention include Peregal, alkylphenol polyoxyethylene ether, fatty acid polyoxyethylene ether, polyol surfactant, span and polyethers. In some preferred embodiments, the polyoxyethylene nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride.

[0124] The content of the water-soluble polyol can be 40 to 67 parts, for example 45, 55, or 60 parts. In some embodiments, the water-soluble polyol includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

[0125] In some embodiments, the weight average molecular weight of the polyethylene glycol is 200-1000, for example, 300, 400, 500, 600, 700, 800, or 900.

[0126] In some embodiments, the solvent has an HLB value of 9 to 29, such as 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, and 29.

[0127] In some embodiments, the coenzyme Q 10 A solution is a clear solution. The term "clear solution" is used to distinguish it from liquid preparations such as suspensions or emulsions.

[0128] The present invention also relates to a liquid composition comprising the above-mentioned coenzyme Q 10 Solutions and cardioplegic solutions, wherein the concentration of the dissolving agent in the cardioplegic solution is below 10 mg / mL, for example 9 mg / mL, 8 mg / mL, 7 mg / mL, 6 mg / mL, 5 mg / mL, 4 mg / mL, 3 mg / mL, 2 mg / mL, 1 mg / mL; preferably 1.1 mg / mL to 10 mg / mL.

[0129] In some embodiments, the composition further contains thiourea, and the concentration of thiourea is preferably 0.06 mg / mL to 0.54 mg / mL, for example, 0.10 mg / mL, 0.15 mg / mL, 0.20 mg / mL, 0.25 mg / mL, 0.30 mg / mL, 0.35 mg / mL, 0.40 mg / mL, 0.45 mg / mL, and 0.50 mg / mL.

[0130] In some embodiments, the composition contains coenzyme Q 10 The concentration is 15 mg / L to 60 mg / L, for example, 17 mg / L, 20 mg / L, 23 mg / L, 25 mg / L, 27 mg / L, 30 mg / L, 35 mg / L, 40 mg / L, 45 mg / L, 50 mg / L, and 55 mg / L.

[0131] A cardioplegic solution (abbreviated as "cardioplegic solution") is a liquid that is infused into the heart via the coronary artery or coronary sinus during open-heart surgery, chemically inducing a rapid cardiac arrest. Cardioplegic solutions suitable for use in the present invention are preferably intracellular solutions, more preferably low-sodium and low-calcium solutions, for example, with a sodium ion concentration below 15 mEq / L and a calcium ion concentration below 0.015 mEq / L. More preferably, the cardioplegic solution contains tryptophan and ketoglutarate ions, and uses histidine as a buffer solution. In some specific embodiments, the cardioplegic solution is HTK solution (the histidine-tryptophan-ketoglutarate solution; Custodiol, also known as Custodiol).

[0132] The liquid compositions of the present invention may also contain other ingredients as desired. In some cases, it may be beneficial for the liquid composition to contain a thrombolytic agent, such as tissue plasminogen activator, as well as other free radical scavengers or drugs that prevent the generation of free radicals. Examples of free radical scavengers include superoxide dismutase, a protein that is expressed after an ischemic event, or less potent substances such as catalase, acetylcysteine, vitamin E, glutathione, and selenium.

[0133] According to yet another aspect of the present invention, it also relates to the use of the above composition in the preservation of an isolated donor heart in heart transplantation.

[0134] The preferred storage temperature is about 0° C. to about 35° C., preferably about 0° C. to about 16° C., and more preferably about 0° C. to about 4° C. Generally speaking, at low temperatures, the time an isolated organ can withstand ischemia can be extended 10 times longer than at room temperature.

[0135] According to another aspect of the present invention, it also relates to the use of the above-mentioned composition in the preparation of a cardioplegic solution for perfusion during establishment of extracorporeal circulation.

[0136] According to yet another aspect of the present invention, there is also provided a method for inducing temporary cardiac paralysis / establishing extracorporeal circulation during cardiac surgery, comprising the step of administering an effective amount of the liquid composition as described above to perfuse the patient's heart.

[0137] As used herein, the term "effective amount" or "therapeutically effective amount" of a drug or pharmaceutically active ingredient is used to refer to a non-toxic amount of the drug or active agent that is sufficient to provide the desired therapeutic effect. An "effective" amount will vary from individual to individual, depending on the individual's age and general condition, the specific active agent, and the like. An appropriate "effective" amount in any particular case can be determined by one skilled in the art using routine experimentation.

[0138] In some embodiments, the liquid composition is administered to the patient at low temperature.

[0139] In some embodiments, the liquid composition is administered to the patient at room temperature.

[0140] In some embodiments, the liquid composition is administered only once during surgery.

[0141] In some embodiments, the liquid composition is administered more than once during surgery.

[0142] In some embodiments, the liquid composition is infused into the root of a cross-clamped aorta and / or directly into the coronary sinus. A balloon catheter can be used as a conduit for the liquid composition, which is introduced through the right atrium into the coronary sinus and then infused into the coronary circulation via the venous circulation. This approach has the advantage of allowing for more uniform distribution of the liquid composition in patients with diffuse coronary artery disease, and its delivery does not rely on a functioning aortic valve.

[0143] In some embodiments, the method of using the liquid composition of the present invention to induce temporary cardiac paralysis during cardiac surgery comprises perfusing the patient's heart with the liquid composition of the present invention at a temperature of about 4°C to about 35°C, preferably at a temperature of about 10°C to about 21°C, and most preferably at about 13°C. It is known to those skilled in the art that the process of using the present invention to induce cardiac paralysis in a patient to preserve cardiac function comprises perfusing the heart with the liquid composition under moderate hypothermia. The administration process of the present invention further comprises perfusing the heart with the liquid composition, preferably at a temperature of about 20 to about 30 mL / kg. In some embodiments, the process comprises perfusing the patient's heart with the liquid composition of the present invention at a moderate hypothermia every 20 to 40 minutes, inducing cardiac paralysis to preserve cardiac function. The method of use of the present invention may comprise perfusing the heart with the liquid composition of the present invention in a cycle of about 20 to 40 minutes, continuing at a moderate hypothermia for at least about 24 cycles. The method of use of the present invention comprises continuously administering the liquid composition.

[0144] Myocardial ischemia-reperfusion injury in patients undergoing cardiac surgery after extracorporeal circulation can severely damage myocardial tissue activity and affect postoperative cardiac function. Our previous studies have shown that supplementing cardioplegia with additional coenzyme Q 10 It can enhance the myocardial protection effect during extracorporeal circulation and reduce postoperative ischemia-reperfusion injury.

[0145] The embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. For experimental methods in the following examples where specific conditions are not specified, reference is made to the guidance provided in the present invention, and may also be made to experimental manuals or conventional conditions in the art, other experimental methods known in the art, or conditions recommended by the manufacturer.

[0146] In the following specific examples, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument testing accuracy or operational accuracy are allowed.

[0147] Example 1

[0148] Coenzyme Q 10 Solution preparation:

[0149] Formula 1. Weigh 400mg of polyoxyethylene hydrogenated castor oil (Cremophor RH40), 600mg of glycerol, heat to 40℃, add coenzyme Q 10 20 mg, vortex and mix evenly to obtain a clear solution with an HLB value of 14-16.

[0150] Formulation 2: Weigh 600 mg of polyoxyethylene 12-hydroxystearic acid (Kolliphor HS15), 400 mg of polyethylene glycol 400, heat to 40°C, and add coenzyme Q 10 40 mg, vortex and mix well to obtain a clear solution with an HLB value of 16.

[0151] Formula 3. Weigh 200mg of Labrafil M1944 (Oleoyl polyoxyl-6 glycerides), 400mg of glycerol, and add coenzyme Q 10 15 mg, heat in a 50°C water bath for about 5 min, mix well, and obtain a clear solution with an HLB value of 9.

[0152] Formulation 4. Weigh 400 mg of Labrafil M 2130 (Lauroyl polyoxyl-6 glycerides) and 600 mg of glycerol, heat to 50°C, and add coenzyme Q 10 15 mg, mix well to obtain a clear solution with an HLB value of 9.

[0153] Formula 5. Weigh 400 mg of Cremophor EL, 400 mg of glycerol, and add coenzyme Q 10 50 mg, heat in a 50°C water bath for about 5 min, mix well, and obtain a clear solution with an HLB value of 13.5.

[0154] Formula 6. Weigh 200 mg of Labrosol (Caprylocaproyl Polyoxyl-8 glycerides), 400 mg of polyethylene glycol 300, and add coenzyme Q 10 15 mg, heat in a 50°C water bath for about 5 min, mix well, and obtain a clear solution with an HLB value of 12.

[0155] Formulation 7. Weigh 180 mg of Pluronic F68 and 520 mg of polyethylene glycol 300, heat and melt; add coenzyme Q 10 20 mg, mixed to obtain a translucent clear solution with an HLB value of 29.

[0156] The main properties of formula 1 to formula 7 are shown in Table 1:

[0157] Table 1

[0158] As shown in Figure 1A and B, the coenzyme Q prepared by formula 1 and formula 2 10The solution is a yellow, transparent, viscous solution. After diluting the solution with cardioplegic solution to about 60 μg / mL, a clear, light yellow solution is obtained (Formulas 1 and 2 in Figure 1C). In addition, the coenzyme Q prepared in Formula 3 10 The solution obtained after dilution is also clear and light yellow. 10 The solution is translucent in appearance.

[0159] Example 2

[0160] Contains coenzyme Q 10 Particle size determination of cardioplegic solutions

[0161] Take the coenzyme Q of formula 1, 2 and 7 in Example 1 10 The solution was diluted to 60 μg / mL with cardioplegic solution. The average particle size and particle size distribution were determined using dynamic light scattering (Zetasizer Nano ZS90, Malvern, UK). The laser wavelength was 633 nm, the temperature was 25°C, and the equilibrium time was 120 seconds. The measurements were performed according to the viscosity and refractive index of water. The results are shown in Table 2. 10 The particle size determination results of the solution in the cardioplegic solution are shown in Table 2:

[0162] The volume average particle size distribution of the three samples is shown in Figure 2. Figures 2A to 2C are samples prepared from Formula 1, Formula 2, and Formula 7 in Example 1, respectively.

[0163] The results of particle size determination showed that during dilution with cardioplegia solution, coenzyme Q 10 It self-assembles with polyoxyethylene surfactants to form micelles, which 10 Effective solubilization in micelles prevents drug precipitation. The micelle particle size distribution is relatively uniform. The Z-average particle size of Formulation 7 is over 26 times larger than that of the other formulations, which may explain the lower clarity of Formulation 7.

[0164] Example 3

[0165] Toxicity assessment of solvents and reducing agents

[0166] The dissolving agent in this embodiment refers to the dissolving coenzyme Q 10 Solvents, including nonionic surfactants and water-soluble polyols

[0167] 1. Obtaining Cardiomyocytes

[0168] Human induced pluripotent stem cells were cultured in vitro and induced to differentiate into cardiomyocytes. Flow cytometry was used to assess differentiation efficiency, and patch clamp electrophysiological activity was used to verify cardiomyocyte differentiation.

[0169] In Figure 3, Figure 3A selects the live cell population P1 based on forward scattered light and side scattered light, and removes dead cells; Figure 3B selects the cell population P2 located on the diagonal line based on the height and area of ​​forward scattered light, and removes the sticky cell population; Figure 3C circles the control cell population that is not labeled with cardiac troponin T antibody outside the gate; Figures 3D and E identify the cardiac troponin T-positive cardiomyocyte population P3 based on the gate in C, and flow cytometry detection shows that the cardiac troponin T-positive cardiomyocytes account for 85.9%; Figure 3F shows the action potential of human induced pluripotent stem cell-derived cardiomyocytes, which can always maintain strong and stable spontaneous action potentials.

[0170] Figures 3A-E show flow cytometry analysis of human induced pluripotent stem cell-derived cardiomyocytes: The positive rate for the cardiomyocyte-specific marker troponin T was 81.28% ± 2.41% (n = 5), demonstrating that this protocol can yield a large number of highly pure cardiomyocytes. Figure 3F shows that the cardiomyocytes consistently maintain robust and stable spontaneous action potentials, demonstrating that the cardiomyocytes obtained using this protocol can function normally.

[0171] 2. Evaluation and optimization of cytotoxicity of dissolving agents

[0172] 400 mg of Cremophor RH40 and 600 mg of glycerol were weighed, heated to 40°C, and vortexed to mix thoroughly to obtain a clear mixed solution. 1000 mg, 330 mg, and 110 mg of the mixed solution were added to 100 mL of HTK cardioplegia solution, respectively, to prepare HTK cardioplegia solutions containing 10 mg / mL, 3.3 mg / mL, and 1.1 mg / mL of the soluble agent. Cardiomyocytes were then cultured in the cardioplegia solutions containing different concentrations of the soluble agent for 30 minutes. Western blot analysis was used to detect activation of the cardiomyocyte apoptosis pathway, and TUNEL staining was used to detect the apoptosis rate. The toxicity of different concentrations of the soluble agent on cardiomyocytes was evaluated to explore the safe concentration range of the soluble agent.

[0173] Figure 4A shows the Western blot analysis of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) in cardiomyocytes cultured in HTK cardioplegia solution containing 10 mg / mL, 3.3 mg / mL, and 1.1 mg / mL of the lytic agent. Figure 4B is a bar graph showing activation of the apoptosis pathway by Western blot. The results showed that compared to the negative control (HTK cardioplegia solution without any other components), the addition of 10 mg / mL, 3.3 mg / mL, and 1.1 mg / mL of the lytic agent to the cardioplegia solution did not result in statistically significant differences in cardiomyocyte apoptosis. Figure 4C shows TUNEL staining of cardiomyocytes in cardioplegia containing different concentrations of the lytic agent. C1-C4, C5-C8, C9-C12, and C13-C16 show TUNEL staining of cardiomyocytes in HTK cardioplegia containing 10 mg / mL, 3.3 mg / mL, and 1.1 mg / mL of the lytic agent, respectively. DAPI (nuclei), TUNEL (apoptotic cells), and Actinin (cytoskeleton) are shown. The scale bar is 50 μm. Figure 4D is a bar graph of TUNEL staining for cardiomyocyte apoptosis. The results showed that compared with the negative control group, the addition of 10 mg / mL, 3.3 mg / mL, and 1.1 mg / mL of the lytic agent to the cardioplegia significantly reduced the apoptosis rate of cardiomyocytes, with no statistical difference between the groups.

[0174] From the results in FIG4 , it can be seen that when the concentration of the dissolving agent is below 10 mg / mL, there is no significant effect on the apoptosis level of cardiomyocytes, that is, when the concentration of the dissolving agent is below 10 mg / mL, there is no cardiomyocyte toxicity.

[0175] 3. Evaluation and Optimization of Reducing Agent Toxicity to Cardiomyocytes

[0176] Thiourea (54 mg, 18 mg, and 6 mg) was weighed and evenly dissolved in 100 mL of HTK cardioplegia solution to prepare cardioplegia solutions with concentrations of 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL. Cardiomyocytes were cultured in these solutions for 30 minutes. The activation level of the myocardial apoptosis pathway was detected by Western blot, and the apoptosis rate of myocardial cells was detected by TUNEL staining. The toxicity of different concentrations of reducing agents on myocardial cells was evaluated to explore the safe concentration range of reducing agents.

[0177] Figure 5A shows the Western blot analysis of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) in cardiomyocytes cultured in HTK cardioplegia solution containing different concentrations of the reducing agent thiourea (0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL). Figure 5B is a bar graph showing activation of the apoptosis pathway by Western blot. The results showed that compared to the negative control (HTK cardioplegia solution without any other components), the addition of 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL of the reducing agent to the cardioplegia solution did not result in statistically significant differences in cardiomyocyte apoptosis. Figure 5C shows TUNEL staining of cardiomyocytes in cardioplegia solutions containing different concentrations of reducing agents. C1-C4, C5-C8, C9-C12, and C13-C16 show TUNEL staining of cardiomyocytes after incubation in HTK cardioplegia solutions containing 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL thiourea, respectively. DAPI (nuclei), TUNEL (apoptotic cells), and actinin (cytoskeleton) are shown. The scale bar is 50 μm. Figure 5D is a bar graph of TUNEL staining for cardiomyocyte apoptosis. The results showed that compared with the negative control group, the addition of 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL reducing agents to the cardioplegia solution significantly reduced the apoptosis rate of cardiomyocytes, with no statistical difference between the groups.

[0178] From the results in FIG5 , it can be seen that thiourea concentrations below 0.54 mg / mL have no significant effect on the apoptosis rate of cardiomyocytes, that is, thiourea concentrations below 0.54 mg / mL have no cardiomyocyte toxicity.

[0179] Example 4

[0180] Coenzyme Q 10 Solution stability assessment

[0181] According to the recipe 1 of Example 1, 400 mg of polyoxyethylene hydrogenated castor oil (Cremophor RH40) and 600 mg of glycerol were weighed and heated to 40° C., and coenzyme Q was added. 10 20 mg, vortex and mix evenly to obtain clear coenzyme Q 10 Solution; weigh 306mg coenzyme Q 10 The solution was added to 100 ml of HTK cardioplegia solution and stirred until uniformly mixed to prepare 60 mg / L coenzyme Q 10 cardioplegia solution;

[0182] Then, 54 mg, 18 mg, and 6 mg of thiourea were weighed and dissolved in 100 mL of the above coenzyme Q 10Three new cardioplegic solutions with different concentrations of thiourea were prepared from the cardioplegic solution: 0.54 mg / mL, 0.18 mg / mL, and 0.06 mg / mL.

[0183] Finally, the prepared solution was stored in a sealed container at 4°C away from light. After 0, 12, 15, 18, 21, 24, 27 and 30 days, the coenzyme Q in the three new cardioplegic solutions containing different concentrations of thiourea and the new cardioplegic solution without thiourea was detected by high performance liquid chromatography. 10 At the same time, the contents of α-ketoglutarate, tryptophan and histidine were detected by colorimetric ketoglutarate quantitative kit and ultra-performance liquid chromatography tandem mass spectrometry on the 0th, 3rd, 6th, 9th, 12th, 15th, 18th, 21st, 24th, 27th and 30th days respectively, and the contents of solubilizer, coenzyme Q 10 , whether the addition of reducing agents affects the stability of HTK cardioplegia solution, and evaluate the effect of different concentrations of thiourea as a reducing agent on coenzyme Q 10 Stability protection effect.

[0184] Figure 6: Figures 6A, B, C, and D are respectively the coenzyme Q in the new cardioplegia solution containing 0.54 mg / mL, 0.18 mg / mL, 0.06 mg / mL thiourea and no thiourea. 10 6E, F, and G are curves showing the changes in the contents of α-ketoglutarate, tryptophan, and histidine over time in the new cardioplegia solution containing 0.54 mg / mL thiourea.

[0185] The results in Figure 6A, B, C, and D show that the coenzyme Q 10 The concentration of coenzyme Q 10 The concentration decreased significantly within 30 days, indicating that coenzyme Q 10 The stability is poor without reducing agent protection, while 0.54 mg / mL, 0.18 mg / mL and 0.06 mg / mL thiourea can play a good role in coenzyme Q 10 Therefore, in order to ensure the protection of coenzyme Q 10 To ensure stability and reduce the amount of thiourea added, the optimal concentration of thiourea in cardioplegia solution is 0.06 mg / L.

[0186] The results in Figure 6E, F, and G showed that the contents of α-ketoglutarate, tryptophan, and histidine did not decrease significantly within 30 days, indicating that the dissolution agent and coenzyme Q in the new cardioplegia solution 10 The addition of reducing agents will not disturb the main components of HTK cardioplegia solution, and the solution is highly stable.

[0187] Example 5

[0188] Contains coenzyme Q 10 Evaluation of the cardioplegic effect of cardioplegic solution composition

[0189] Weigh 54 mg of thiourea and dissolve it evenly in 100 mL of HTK cardioplegia solution to prepare 0.54 mg / mL HTK cardioplegia-containing solution.

[0190] According to the recipe 1 of Example 1, 400 mg of polyoxyethylene hydrogenated castor oil (Cremophor RH40) and 600 mg of glycerol were weighed and heated to 40° C., and coenzyme Q was added. 10 20 mg, vortex and mix evenly to obtain clear coenzyme Q 10 Solution, then, weigh 306 mg of coenzyme Q 10 The solution was added to 100 ml of HTK cardioplegia solution and stirred until uniformly mixed to prepare 60 mg / L coenzyme Q 10 Composition of HTK cardioplegia solution.

[0191] Weigh 250 mg of sodium bisulfite and evenly dissolve it in 100 mL of HTK cardioplegia solution to prepare 2.5 mg / mL HTK cardioplegia solution containing sodium bisulfite.

[0192] The above HTK cardioplegia solution containing thiourea and coenzyme Q 10 Cardiomyocytes were perfused with HTK cardioplegia solution composition, HTK cardioplegia solution containing sodium bisulfite, and HTK cardioplegia solution without any other ingredients, and the action potential of cardiomyocytes was detected by cell patch clamp technique. The differences in cardioplegia effects among the four groups were compared.

[0193] Figure 7A, B, C, and D are myocardial cells perfused with HTK cardioplegia solution, HTK cardioplegia solution containing 0.54 mg / mL thiourea, and 60 mg / L coenzyme Q 10 The action potential of the myocardial cells after treatment with HTK cardioplegia solution and HTK cardioplegia solution containing 2.5 mg / mL sodium bisulfite. The red arrows indicate that the electrophysiological activity of the myocardial cells stops, and the blue arrows indicate that the action potential of the myocardial cells restarts.

[0194] The results in Figure 7 show that myocardial cells stopped action potential almost simultaneously after perfusion of the four groups of cardioplegia solutions. Groups A, B, and C in Figure 7 all recovered action potential within 11 minutes, while group D in Figure 7 did not recover action potential within 45 minutes. 10 The addition of sodium bisulfite had no significant effect on the cardioplegic effect of the cardioplegic solution, but the myocardial cell re-beating time was significantly prolonged after the addition of sodium bisulfite.

[0195] Example 6

[0196] Contains coenzyme Q 10 Evaluation of the myocardial protective effect of cardioplegic solution compositions

[0197] 1. According to the recipe 1 of Example 1, weigh 400 mg of polyoxyethylene hydrogenated castor oil (Cremophor RH40), 600 mg of glycerol, heat to 40°C, and add coenzyme Q 10 20 mg, vortex and mix evenly to obtain clear coenzyme Q 10 Then, 76.5 mg, 153 mg, 229.5 mg, and 306 mg of the obtained coenzyme Q were weighed respectively. 10 The solution was added to 100 ml of HTK cardioplegia solution and stirred until uniformly mixed. Four different concentrations of coenzyme Q were prepared, namely 15 mg / L, 30 mg / L, 45 mg / L and 60 mg / L. 10 Composition of cardioplegic solution.

[0198] First, myocardial cells were placed in a hypoxic environment with different concentrations of coenzyme Q 10 The myocardial cells were cultured in a HTK cardioplegia solution composition and a HTK cardioplegia solution without any other ingredients for 45 minutes to simulate myocardial ischemia. Then, the myocardial cells were cultured in a high oxygen environment and a normal cell culture medium for 3 hours to simulate myocardial reperfusion. The activation level of the myocardial cell apoptosis pathway was detected by Western blot and the apoptosis rate of myocardial cells was detected by TUNEL staining to evaluate the effect of different concentrations of coenzyme Q 10 Protective effect of coenzyme Q on cardiomyocytes 10 The optimal dosage.

[0199] Figure 8A shows myocardial cells in HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L and 60 mg / L coenzyme Q 10 Figure 8B is a bar graph of Western blot detection of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) in HTK cardioplegia solution after ischemia-reperfusion. Figure 8C is a TUNEL staining image of cardiomyocytes, where C1-C4, C5-C8, C9-C12, C13-C16, and C17-C20 are the activation levels of cardiomyocytes in HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L, and 60 mg / L coenzyme Q, respectively. 10 Figure 8D is a bar graph of the apoptosis rate of myocardial cells detected by TUNEL staining.

[0200] Figure 8A-B shows that compared with HTK cardioplegia solution without any other ingredients, the activation level of myocardial cell apoptosis signaling pathway was significantly increased in the presence of coenzyme Q 10 It starts to decrease when the concentration is 15mg / L and decreases significantly when the concentration is 30mg / L. 10 As the concentration continued to increase, the apoptosis level did not change significantly. Figure 8C-D shows that compared with HTK cardioplegia solution without any other ingredients, the apoptosis rate of myocardial cells increased with the addition of coenzyme Q 10 The concentration began to decrease when it was 15 mg / L. 10 When the concentration is 30mg / L, it decreases significantly. 10 When the concentration continued to increase, the apoptosis rate of myocardial cells did not decrease significantly.

[0201] 2. According to the recipe 2 of Example 1, 600 mg of polyoxyethylated 12-hydroxystearic acid (Kolliphor HS15) and 400 mg of polyethylene glycol 400 were weighed and heated to 40°C. 40 mg of coenzyme Q was added. 10 , vortex and mix evenly to obtain clear coenzyme Q 10 Mix the solution; then weigh 39mg, 78mg, 117mg, and 156mg of coenzyme Q 10 The mixed solution was added to 100 ml of HTK cardioplegia solution and stirred until uniformly mixed. Four different concentrations of coenzyme Q were prepared, namely 15 mg / L, 30 mg / L, 45 mg / L and 60 mg / L. 10 Composition of HTK cardioplegia solution.

[0202] First, myocardial cells were placed in a hypoxic environment with different concentrations of coenzyme Q 10 The myocardial cells were cultured in a HTK cardioplegia solution composition and a HTK cardioplegia solution without any other ingredients for 45 minutes to simulate myocardial ischemia. Then, the myocardial cells were cultured in a high oxygen environment and a normal cell culture medium for 3 hours to simulate myocardial reperfusion. The activation level of the myocardial cell apoptosis pathway was detected by Western blot and the apoptosis rate of myocardial cells was detected by TUNEL staining to evaluate the effect of different concentrations of coenzyme Q 10 Protective effect of coenzyme Q on cardiomyocytes 10 The optimal dosage.

[0203] Figure 9A shows myocardial cells in HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L and 60 mg / L coenzyme Q 10Figure 9B is a bar graph showing the activation level of apoptosis pathway detected by Western blot. Figure 9C is a TUNEL staining image of cardiomyocytes, where C1-C4, C5-C8, C9-C12, C13-C16, and C17-C20 are the activation levels of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) in HTK cardioplegia solution after ischemia-reperfusion. Figure 9C is a TUNEL staining image of cardiomyocytes, where C1-C4, C5-C8, C9-C12, C13-C16, and C17-C20 are the activation levels of apoptosis signaling pathway proteins (Caspase-3 and Cleaved Caspase-3) in HTK cardioplegia solution containing 15 mg / L, 30 mg / L, 45 mg / L, and 60 mg / L coenzyme Q 10 Figure 9D is a bar graph of the apoptosis rate of myocardial cells detected by TUNEL staining.

[0204] The results in Figure 9A-B show that compared with HTK cardioplegia solution without any other ingredients, the activation level of myocardial cell apoptosis signaling pathway is significantly increased in the presence of coenzyme Q 10 The apoptosis rate of myocardial cells decreased with the addition of coenzyme Q at a concentration of 15 mg / L and decreased significantly with the addition of coenzyme Q at a concentration of 30 mg / L. However, when the coenzyme concentration continued to increase, the apoptosis level did not change significantly. The results in Figure 9C-D show that compared with the HTK cardioplegia solution without any other ingredients, the apoptosis rate of myocardial cells decreased with the addition of coenzyme Q at a concentration of 15 mg / L and decreased significantly with the addition of coenzyme Q at a concentration of 30 mg / L. 10 The concentration began to decrease when it was 15 mg / L. 10 When the concentration is 30mg / L, it decreases significantly. 10 When the concentration continued to increase, the apoptosis rate of myocardial cells did not decrease significantly.

[0205] From the results of Figures 8 and 9, we can see that coenzyme Q 10 When the content of coenzyme Q is 30mg / L, it can achieve good myocardial protection. 10 The optimal concentration is 30 mg / L.

[0206] The advantages of the present invention are further demonstrated by the above examples:

[0207] 1. The coenzyme Q 10 Q prepared by solvent 10 The solution can be well dissolved in HTK stop solution without aggregation and precipitation.

[0208] 2. The coenzyme Q 10 Thiourea can stabilize it in HTK liquid and prevent oxidation and deterioration.

[0209] 3. Contains Coenzyme Q 10The new cardioplegia solution has a protective effect on myocardial cells after ischemia-reperfusion.

[0210] By using solvents and reducing agents, the coenzyme Q 10 The solubility and stability of the cardioplegia solution are difficult to solve. 10 The cardioplegic solution composition is safe and non-toxic, and can significantly enhance the protective effect of the cardioplegic solution on myocardial cells without affecting the cardioplegic effect of the cardioplegic solution.

[0211] The prepared coenzyme Q 10 After the solution is added to the cardioprotective solution, the coenzyme Q 10 The self-assembly of the drug and / or its analogs with surfactants forms micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure. Therefore, only a small amount of surfactant is needed to greatly increase the coenzyme Q in the micelle-like structure. 10 The loading amount is conducive to reducing the amount of excipients in the solution to improve the compatibility of the solution with the cardiac protective solution when used for myocardial protection. At the same time, this preparation method is used to prepare coenzyme Q 10 When loaded, the micelle-like structure dispersed in water also has a limiting effect on particle growth, making coenzyme Q 10 Before the drug can aggregate into large particles, it is effectively loaded into the micelle-like structure in the form of molecules or microcrystals. The drug in the microcrystal state is not only conducive to greatly increasing the drug loading capacity, but also further improves the stability of the drug loading, which can avoid the 10 Precipitation in the cardioprotective solution. 10 The particle size of the solution cannot be measured, that is, coenzyme Q 10 Dispersed in the solution in molecular state, but coenzyme Q 10 After the solution is added to the cardioprotective solution, uniform nanoparticles can be formed.

[0212] The above-described embodiments merely represent several implementation methods of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art may make several modifications and improvements without departing from the concept of the present invention, and these modifications and improvements fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be based on the appended claims, and the description and drawings may be used to interpret the content of the claims.

Claims

1. A coenzyme Q 10 A solution, characterized in that The dissolving agent comprises a pharmaceutically acceptable polyoxyethylene type nonionic surfactant and a water-soluble polyhydroxy compound; The polyoxyethylene type nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride; The water-soluble polyhydroxy compound includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

2. A kind of coenzyme Q 10 The cardioplegic solution composition is characterized in that Contains Coenzyme Q 10 Solutions and cardioplegic solutions, Coenzyme Q 10 A solution, wherein the dissolving agent comprises a pharmaceutically acceptable polyoxyethylene type nonionic surfactant and a water-soluble polyhydroxy compound, and the concentration of the dissolving agent in the cardioplegia solution is not greater than 10 mg / mL; The polyoxyethylene type nonionic surfactant includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride; The water-soluble polyhydroxy compound includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

3. The coenzyme Q-containing composition according to claim 2 10 The cardioplegic solution composition is characterized in that It is any of the following options: Scenario 1: Coenzyme Q 10 A solution, wherein the dissolving agent comprises 33 to 60 parts by weight of a pharmaceutically acceptable polyoxyethylene nonionic surfactant and 40 to 67 parts by weight of a water-soluble polyhydroxy compound, and the concentration of the dissolving agent in the cardioplegia solution is no more than 10 mg / mL; Scenario 2: Coenzyme Q 10 The solution comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant in weight portions of 33 to 60 parts and a water-soluble polyhydroxy compound in weight portions of 40 to 67 parts, coenzyme Q 10 The concentration is 14.8 to 58.8 μg / mg, the concentration of the dissolving agent in the cardioplegia solution is not more than 10 mg / mL, and the coenzyme Q 10 The concentration in the cardioplegia solution is 15 to 60 mg / L; Scenario 3: Coenzyme Q 10 The solution comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant in weight portions of 33 to 60 parts and a water-soluble polyhydroxy compound in weight portions of 40 to 67 parts, coenzyme Q 10 The concentration is 14.8 to 58.8 μg / mg, and the obtained solution is a clear solution with an HLB value of 9 to 29. The concentration of the solvent in the cardioplegia solution is not more than 10 mg / mL, and the coenzyme Q 10 The concentration in the cardioplegia solution is 15 to 60 mg / L; Solution 4: Coenzyme Q 10 The solution comprises a pharmaceutically acceptable polyoxyethylene nonionic surfactant in weight portions of 33 to 60 parts and a water-soluble polyhydroxy compound in weight portions of 40 to 67 parts, coenzyme Q 10 The concentration is 14.8-58.8 μg / mg, and the resulting solution is a clear solution with an HLB value of 9-29. 10 The self-assembly with the surfactant forms micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure; the concentration of the dissolving agent in the cardioplegia solution is not more than 10 mg / mL, and the coenzyme Q 10 The concentration in the cardioplegia solution is 15 to 60 mg / L; The polyoxyethylene type nonionic surfactant as a solubilizing agent includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride; The water-soluble polyhydroxy compound includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol; Solution 5: Coenzyme Q 10 The solution comprises 33 to 60 parts by weight of a pharmaceutically acceptable polyoxyethylene nonionic surfactant as a solubilizer and 40 to 67 parts by weight of a water-soluble polyhydroxy compound as a cosolvent, and coenzyme Q 10 The concentration is 14.8-58.8 μg / mg, and the resulting solution is a clear solution with an HLB value of 9-29. 10 The self-assembly with the surfactant forms micelle-like structure particles, so that the drug loading process is always carried out in the nano-scale micelle-like structure; the concentration of the dissolving agent in the cardioplegia solution is not more than 10 mg / mL, and the coenzyme Q 10 The concentration is 15-60 mg / L; The polyoxyethylene type nonionic surfactant as a solubilizing agent includes one or more of polyoxyethylene hydrogenated castor oil, polyoxyethylene 12-hydroxystearic acid, caprylic capric macrogol glyceride, poloxamer, lauroyl polyoxyethylene-6 glyceride, polyoxyethylene castor oil, polyoxyethylene-8 behenic acid glyceride, lauroyl polyoxyethylene-32 glyceride, stearoyl polyoxyethylene glyceride, oleoyl polyoxyethylene glyceride and linoleoyl polyoxyethylene-6 glyceride; The water-soluble polyhydroxy compound as a cosolvent includes one or more of glycerol, polyethylene glycol, polyglycerol, a copolymer of polyethylene glycol and polypropylene glycol, and polypropylene glycol.

4. The coenzyme Q-containing composition according to claim 2 or 3 10 The cardioplegic solution composition is characterized in that The solubilizer and cosolvent in the dissolving agent are mixed with coenzyme Q at 40-50°C. 10 Mix well to obtain coenzyme Q 10 Solution, the resulting coenzyme Q 10 The solution exists in a semi-solid form at low temperatures. After being heated in a water bath, it returns to a clear solution and is added to the cardiac protective solution. It self-assembles with the surfactant to form a solution with coenzyme Q encapsulated inside. 10 The micelle-like self-assembled structure makes coenzyme Q 10 Stably dispersed in the cardioprotective solution.

5. The coenzyme Q-containing composition according to claim 2 or 3. 10 The cardioplegic solution composition is characterized in that The weight average molecular weight of the polyethylene glycol is 200-1000.

6. The coenzyme Q-containing composition according to claim 2 or 3. 10 The cardioplegic solution composition is characterized in that The concentration of the dissolving agent in the cardioplegia solution is 1.1 mg / mL to 10 mg / mL, Alternatively, the dissolving agent comprises 40 parts, 60 parts, 33.3 parts or 50 parts by weight of a polyoxyethylene nonionic surfactant as a solubilizing agent; Alternatively, the HLB value is between 9, 14, 16, 13.5 or 12; Alternatively, the coenzyme Q 10 In the coenzyme Q 10 The concentration in the solution is 14.8 to 58.8 μg / mg, such as 19.6 μg / mg, 38.5 μg / mg, 24.3 μg / mg, 14.8 μg / mg, 58.8 μg / mg or 24.4 μg / mg; Alternatively, the coenzyme Q 10 The concentration in the cardioplegic solution is 15 mg / L, 17 mg / L, 20 mg / L, 23 mg / L, 25 mg / L, 27 mg / L, 30 mg / L, 35 mg / L, 40 mg / L, 45 mg / L, 50 mg / L, 55 mg / L or 60 mg / L.

7. The coenzyme Q-containing composition according to claim 2 or 3. 10 The cardioplegic solution composition is characterized in that Coenzyme Q 10 The concentration is 15-30 mg / L; Alternatively, the solubilizers are all non-ionic surfactants, with polyethylene glycol structure as the hydrophilic group, and the HLB value is between 9-29; the cosolvent is a polyhydroxy hydrophilic component, and the cosolvent is polyethylene glycol 300, 400 or glycerol.

8. The coenzyme Q-containing composition according to claim 2 or 3. 10 The cardioplegic solution composition is characterized in that It also contains thiourea at a concentration of 0.06 mg / mL to 0.54 mg / mL.

9. The coenzyme Q-containing composition according to claim 2 or 3. 10 The cardioplegic solution composition is characterized in that The cardioplegic solution is an intracellular fluid type cardioplegic solution.

10. The coenzyme Q-containing composition according to claim 9. 10 The cardioplegic solution composition is characterized in that The cardioplegic solution is HTK cardioplegic solution.

11. The coenzyme Q-containing composition according to claim 2 or 3. 10 The cardioplegic solution composition is characterized in that The coenzyme Q 10 The raw materials of the solution are any of the following formulas: Formula a. Polyoxyethylene hydrogenated castor oil, glycerin and coenzyme Q 10 ; Formulation b. Polyoxyethylated 12-hydroxystearic acid, polyethylene glycol 400 and coenzyme Q 10 ; Formula c. Oleoyl polyoxyethylene glyceride, glycerol and coenzyme Q 10 ; Formula d. Lauroyl polyoxyethylene-6 glyceride, glycerin and coenzyme Q 10 ; Formula e. Polyoxyethylene castor oil, glycerin and coenzyme Q 10 ; Formula f. Caprylic / capric acid macrogol glyceride, polyethylene glycol 300 and coenzyme Q 10 .

12. The coenzyme Q-containing composition according to claim 11. 10 The cardioplegic solution composition is characterized in that The coenzyme Q 10 The raw materials of the solution are any of the following formulas: Formula 1. 400mg polyoxyethylene hydrogenated castor oil, 600mg glycerin and 20mg coenzyme Q 10 ; Formula 2. 600 mg polyoxyethylated 12-hydroxystearic acid, 400 mg polyethylene glycol 400 and 40 mg coenzyme Q 10 ; Formula 3. 200mg oleoyl polyoxyethylene glyceride, 400mg glycerol and 15mg coenzyme Q 10 ; Formula 4. 400mg lauroyl polyoxyethylene-6 glyceride, 600mg glycerin and 15mg coenzyme Q 10 ; Formula 5. 400mg polyoxyethylene castor oil, 400mg glycerin and 50mg coenzyme Q 10 ; Formula 6. 200 mg caprylic / capric macrogol glycerides, 400 mg polyethylene glycol 300 and 15 mg coenzyme Q 10 .

13. Use of the composition according to any one of claims 2 to 12 in the preservation of an isolated donor heart in heart transplantation.

14. Use of the composition according to any one of claims 2 to 12 in the preparation of a cardioplegia solution for perfusion during establishment of extracorporeal circulation.

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