A method for preparing an alpha-ketoglutarate-containing cardioplegic solution

Abstract

The application relates to the technical field of organ protection, in particular to a preparation method of an alpha-ketoglutarate-containing myocardial arrest solution. The method discards the traditional one-off mixing process, adopts a strategy of separately preparing an alpha-ketoglutarate-containing solution and a histidine compound-containing basic solution, and avoids degradation reactions of the two from the source. The core lies in separately preparing the alpha-ketoglutarate-containing solution and the histidine compound solution, filling the two solutions into different chambers of a multi-chamber packaging container or different containers, and preferably independently performing deep oxygen removal treatment on the alpha-ketoglutarate-containing solution to make the dissolved oxygen content of the alpha-ketoglutarate-containing solution be less than or equal to 3.0 ppm. The application solves the problems of oxidative degradation of alpha-ketoglutarate and carbamoyl condensation reaction with histidine compounds by a step-by-step and zoned preparation concept, the prepared product has super-normal chemical stability, supports normal-temperature storage and an ultra-long effective period, significantly reduces production cost and logistics burden, and ensures the accuracy and reliability of a clinical drug dosage.

Description

Technical Field

[0001] This application relates to the field of organ protection technology, and in particular to a method for preparing a myocardial arrest solution containing α-ketoglutarate. Background Technology

[0002] In cardiac surgery, hypothermic cardioplegic solutions are often used to maximize the protection of cardiomyocyte function during cardiac arrest. Among these, histidine-tryptophan-ketoglutarate (HTK) cardioplegic solution is widely used due to its excellent cardioprotective effect. The core component of HTK solution includes α-ketoglutarate, which serves as an energy substrate and metabolic intermediate. α-Ketoglutarate not only participates in the tricarboxylic acid cycle, providing limited energy support to ischemic myocardium, but more importantly, it acts as a cofactor for various dioxygenases, playing a crucial role in the adaptive regulation of cells during hypoxia / reoxygenation and in mitigating oxidative stress damage.

[0003] However, the chemical stability of α-ketoglutarate is a crucial but long-neglected issue in the long-term storage and clinical preparation of HTK cardioplegic solution. Studies have shown that even under recommended low-temperature storage conditions (e.g., 2-8°C), the α-ketoglutarate content in HTK solution still decreases significantly over time; currently marketed products, stored at 2-8°C, have a shelf life of only 12 months. This instability severely impacts the product's shelf life and efficacy, potentially leading to reduced cardioprotective efficacy during clinical use and thus jeopardizing surgical safety.

[0004] Problems with existing technologies and their root causes: Through in-depth research, the inventors discovered that the attenuation of α-ketoglutarate in HTK solution mainly stems from two significant chemical reactions: First, a slow chemical reaction occurs between α-ketoglutarate and another key component in the solution—histidine compounds. The active carbonyl group of α-ketoglutarate may react with the imidazole group and other groups of histidine compounds in a primary stage similar to the Maillard reaction or form an adduct, leading to mutual consumption of the two in a coexisting state.

[0005] Secondly, α-ketoglutarate is sensitive to oxygen and easily oxidized, which reduces its content to some extent. Dissolved oxygen in the solution or oxygen in the headspace air oxidizes α-ketoglutarate, producing byproducts such as succinic acid. This process continues even at low temperatures. Conventional inert gas replacement processes (such as simple nitrogen purging) are insufficient to reduce the dissolved oxygen content in the system to an ideal level, which is usually still above 10 ppm, and cannot effectively prevent the oxidation reaction from continuing for a long time.

[0006] Currently, commercially available HTK cardioplegic solutions are typically packaged in single-chamber bags, pre-mixing all components (including histidine, tryptophan, α-ketoglutarate, etc.) in the same solution. While this packaging method is convenient, it fails to address the stability issues arising from the two aforementioned chemical reactions: it cannot isolate α-ketoglutarate from histidine compounds, nor can it provide a sufficiently low-oxygen environment. Therefore, there is an urgent need in the field for a method that innovates from the source of the preparation process to fundamentally improve the stability of cardioplegic solutions containing α-ketoglutarate. Summary of the Invention

[0007] The purpose of this application is to overcome the shortcomings of existing "one-time mixing and preparation" processes and provide a novel method for preparing α-ketoglutarate-containing cardioplegic solution based on a stepwise preparation concept. The core of this method is to physically separate and differentiate the unstable α-ketoglutarate component from incompatible histidine compounds during the solution preparation stage, thereby interrupting the main degradation pathway at the initial stage of formulation preparation.

[0008] Another objective of this application is to provide a deep deoxygenation process for α-ketoglutarate solution that is compatible with the stepwise preparation method, particularly a technology for creating and maintaining an ultra-low oxygen environment.

[0009] Another objective of this application is to prepare a myocardial arrest solution product with excellent chemical stability through the above-mentioned stepwise preparation and processing technology. This product can be stored at room temperature (or in a cool place), has a significantly extended shelf life, and maintains consistent clinical efficacy.

[0010] In a first aspect, this application provides a method for preparing a myocardial arrest solution containing α-ketoglutarate, using the following technical solution: A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Dissolve the component containing α-ketoglutarate in water for injection; or dissolve the component containing α-ketoglutarate and a pharmaceutically acceptable solute other than histidine compounds in water for injection to form the first solution; S2. Preparation of the second solution: Dissolve the histidine compound in water for injection; or dissolve the histidine compound and a pharmaceutically acceptable solute (excluding α-ketoglutarate) in water for injection to form the second solution. S3. Separate Filling: The second solution and the first solution are separately filled into different chambers of a multi-chamber packaging container, wherein the different chambers are configured to keep the solutions separate after filling and before use; or the second solution and the first solution are separately filled into different containers, for example, the first solution is filled into a glass bottle and the second solution is filled into a soft bag; or the first solution is filled into a glass bottle and the second solution is filled into a glass bottle; or the first solution is filled into a soft bag and the second solution is filled into a soft bag; when in use, the first solution and the second solution are mixed evenly.

[0011] Preferably, before the isolation filling in step S3, the first solution is further subjected to a deep deoxygenation treatment to reduce the dissolved oxygen content.

[0012] Preferably, in step S1, the concentration of α-ketoglutarate in the first solution is 5 to 15 times the target concentration after mixing the first solution and the second solution; the α-ketoglutarate is selected from α-ketoglutarate and / or α-ketoglutarate salt; preferably, the α-ketoglutarate salt is a monosodium α-ketoglutarate salt, wherein the monosodium α-ketoglutarate salt can be selected from a monopotassium α-ketoglutarate salt; the histidine compound is selected from L-histidine and / or L-histidine hydrochloride monohydrate.

[0013] Preferably, the dissolved oxygen content of the first solution is ≤3.0 ppm by performing the deep deoxygenation treatment on the first solution.

[0014] Preferably, the deep deoxygenation treatment includes introducing an inert gas into the first solution, with the following conditions: gas pressure 0.1-0.5 MPa, gas flow rate 0.05-0.5 times the volume of the first solution per minute, and cumulative aeration volume 1-5 times the volume of the first solution. Preferably, in step S3, the isolation filling is carried out under an inert gas protective atmosphere, which is maintained by continuously purging an inert gas curtain, with a purging gas pressure of 0.1-0.3 MPa.

[0015] Preferably, the inert gas is selected from at least one of nitrogen, argon and helium.

[0016] Preferably, the multi-chamber packaging container is a double-chamber soft bag, and the different chambers are isolated from each other by a weak weld.

[0017] Secondly, this application provides a myocardial arrest solution product containing α-ketoglutarate, which adopts the following technical solution: As a general technical concept, this application also provides the above-mentioned α-ketoglutarate myocardial arrest solution product, which is prepared by the above-mentioned method for preparing α-ketoglutarate myocardial arrest solution, and in the final sealed packaging state, the dissolved oxygen content of the first solution is ≤3.0 ppm.

[0018] Thirdly, this application provides a method for preparing a myocardial arrest fluid containing α-ketoglutarate, using the following technical solution: As a general technical concept, this application also provides the use of the above-mentioned method for preparing α-ketoglutarate myocardial arrest solution in the preparation of myocardial protection and / or storage and transportation, especially at room temperature.

[0019] In summary, the beneficial technical effects of this application are as follows: 1. Significant economic benefits: 1) Extended shelf life: By solving the stability problem of α-ketoglutarate, the shelf life of the cardioprotective cardioplegic solution is extended from 12 months to 24 months, reducing the frequency of ordering by users and lowering delivery costs.

[0020] 2) Reduced Transportation Costs: Achieving ambient temperature storage allows cross-border transportation to be switched from air to sea, significantly reducing freight costs. For example, transporting 10 tons of goods (10,000 liters) from Hamburg to Beijing would cost approximately 70,000 euros (570,000 yuan) by air, but only about 20,000 yuan by sea, less than 5% of the air freight cost. Switching 600,000 liters of product to sea freight across borders could save nearly 33 million yuan in transportation costs. This also reduces the need for cold chain transportation, lowers cold chain transportation costs, and increases the options available to business units for choosing transportation companies.

[0021] 2. Product quality improvement: 1) Prevent α-ketoglutarate degradation: Isolate α-ketoglutarate from histidine compounds, create an anaerobic environment to prevent reaction and oxidation of α-ketoglutarate with histidine compounds, ensure product efficacy, and reduce unknown risks. Brief exposure to room temperature during delivery can also reduce the impact on the stability of α-ketoglutarate.

[0022] 2) Improve product consistency: Mix the solution before use to ensure the freshness and accurate proportions of each component of the cardioprotective cardioplegic solution for each use, thereby improving product quality consistency and helping to increase the success rate of cardiac surgery. Detailed Implementation

[0023] The embodiments of this application will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of this application. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products. The α-ketoglutarate ions in the various embodiments of this application are selected from α-ketoglutarate monopotassium salts, and the histidine compounds are selected from L-histidine and L-histidine hydrochloride monohydrate.

[0024] The composition of the α-ketoglutarate myocardial arrest solution used in this application is shown in Table 1: Table 1. Substances and contents of α-ketoglutarate-containing myocardial arrest fluid for use. The composition of the final solution of the myocardial arrest solution containing α-ketoglutarate prepared by the method of this application, after mixing before use, is shown in Table 1.

[0025] Example 1 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of α-ketoglutaric acid monopotassium salt solution with a concentration of 1.842 g / L to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentrations of L-histidine hydrochloride monohydrate, L-histidine, mannitol, sodium chloride, potassium chloride, magnesium chloride hexahydrate, calcium chloride dihydrate, and tryptophan are 4.1926 g / L, 31.0321 g / L, 6.0723 g / L, 0.974 g / L, 0.7456 g / L, 0.9036 g / L, 0.0024 g / L, and 0.4539 g / L. S3. Isolation Filling: Performed under an argon gas protective atmosphere, maintained by continuous purging of an argon gas curtain at a pressure of 0.1 MPa. The second solution and the first solution are filled in equal volumes (9:1 volume ratio) into the two chambers of 100 double-chamber soft-bag packaging containers that have undergone deoxygenation treatment (vacuum nitrogen filling). (The solutions are isolated between the chambers via a weak weld, and the two chambers remain isolated from filling until use.) After filling, heat-sealing clamps instantly melt and seal the bag opening. The sealed double-chamber bag products are sterilized at 115°C for 30 minutes. After sterilization, they are cooled, dried, leak-tested, and labeled to obtain the finished product.

[0026] Example 2 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of a solution of α-ketoglutaric acid monopotassium salt with a concentration of 1.842 g / L and sodium chloride with a concentration of 8.766 g / L to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentrations of L-histidine hydrochloride monohydrate, L-histidine, mannitol, potassium chloride, magnesium chloride hexahydrate, calcium chloride dihydrate, and tryptophan are 4.1926g / L, 31.0321g / L, 6.0723g / L, 0.7456g / L, 0.9036g / L, 0.0024g / L, and 0.4539g / L. S3. Isolation Filling: Conducted under a nitrogen protective atmosphere, maintained by continuous purging of a nitrogen gas curtain at a pressure of 0.3 MPa. The first solution is filled in equal volumes into 100 deoxygenated (vacuum-purified nitrogen-filled) glass bottles, and the second solution is filled in equal volumes into 100 flexible bags. The sealed products are sterilized at 115°C for 30 minutes. After sterilization, the products are cooled, dried, leak-tested, and labeled to obtain the finished product. For use, the glass bottles containing the first solution and the flexible bags containing the second solution are mixed thoroughly.

[0027] Example 3 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of a solution containing 1.842 g / L of α-ketoglutarate monopotassium salt, 54.651 g / L of mannitol, 6.710 g / L of potassium chloride, 8.132 g / L of magnesium chloride hexahydrate, 0.022 g / L of calcium chloride dihydrate, 4.085 g / L of tryptophan, and 8.766 g / L of sodium chloride to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentration of L-histidine hydrochloride monohydrate in the second solution is 4.1926g / L and the concentration of L-histidine is 31.0321g / L; S3. Isolation Filling: Conducted under a helium protective atmosphere, maintained by continuous purging of a helium gas curtain at a pressure of 0.2 MPa. The first solution is filled in equal volumes into 100 deoxygenated (vacuum-filled nitrogen-treated) soft bags, and the second solution is filled in equal volumes into 100 soft bags. The sealed products are sterilized at 115℃ for 30 minutes. After sterilization, the products are cooled, dried, leak-tested, and labeled to obtain the finished product. For use, the soft bags containing the first and second solutions are mixed thoroughly before use.

[0028] Example 4 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of α-ketoglutaric acid monopotassium salt solution with a concentration of 1.842 g / L to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentrations of L-histidine hydrochloride monohydrate, L-histidine, mannitol, sodium chloride, potassium chloride, magnesium chloride hexahydrate, calcium chloride dihydrate, and tryptophan are 4.1926 g / L, 31.0321 g / L, 6.0723 g / L, 0.974 g / L, 0.7456 g / L, 0.9036 g / L, 0.0024 g / L, and 0.4539 g / L. S3. Deep deoxygenation treatment: The first solution is subjected to deep deoxygenation treatment to reduce the dissolved oxygen content. The specific process is to introduce argon gas into the first solution. The gas conditions are: gas pressure 0.1 MPa, gas flow rate 0.5 times the volume of the first solution per minute, and the cumulative gas volume is 1 times the volume of the first solution. S4. Isolation Filling: Conducted under an argon gas protective atmosphere, maintained by continuous purging of an argon gas curtain at a pressure of 0.1 MPa. The second solution and the first solution (after deep deoxygenation treatment) are filled in equal volumes (9:1 volume ratio) into the two chambers of 100 double-chamber soft-bag packaging containers that have undergone deoxygenation treatment (and vacuum nitrogen filling). (The chambers are isolated from each other via a weak weld, maintaining solution isolation between the two chambers from filling until use.) After filling, heat-sealing clamps instantly melt and seal the bag opening. The sealed product is sterilized at 115°C for 30 minutes. After sterilization, it is cooled, dried, leak-tested, and labeled to obtain the finished product. The final dissolved oxygen level of the first solution is measured at 2.8 ppm.

[0029] Example 5 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of α-ketoglutaric acid monopotassium salt solution with a concentration of 1.842 g / L to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentrations of L-histidine hydrochloride monohydrate, L-histidine, mannitol, sodium chloride, potassium chloride, magnesium chloride hexahydrate, calcium chloride dihydrate, and tryptophan are 4.1926 g / L, 31.0321 g / L, 6.0723 g / L, 0.974 g / L, 0.7456 g / L, 0.9036 g / L, 0.0024 g / L, and 0.4539 g / L. S3. Deep deoxygenation treatment: Deep deoxygenation treatment is performed on the first solution to reduce the dissolved oxygen content. The specific process is to introduce helium gas into the first solution. The gas pressure is 0.5 MPa, the gas flow rate is 0.3 times the volume of the first solution per minute, and the cumulative gas volume is 3 times the volume of the first solution. S4. Isolation Filling: Conducted under a helium protective atmosphere, maintained by continuous purging of a helium gas curtain at a pressure of 0.3 MPa. The second solution and the first solution, which has undergone deep deoxygenation treatment, are filled in equal volumes at a volume ratio of 9:1 into the two chambers of 100 double-chamber flexible bag packaging containers that have undergone deoxygenation treatment (vacuum nitrogen filling). (The solutions are isolated between the chambers via a weak weld, and the two chambers remain isolated from filling until use.) After filling, heat-sealing clamps instantly melt and seal the bag opening. The sealed double-chamber bag products are sterilized at 115°C for 30 minutes. After sterilization, they are cooled, dried, leak-tested, and labeled to obtain the finished product. Finally, the dissolved oxygen of the first solution is tested: 2.4 ppm.

[0030] Example 6 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of α-ketoglutaric acid monopotassium salt solution with a concentration of 1.842 g / L to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentrations of L-histidine hydrochloride monohydrate, L-histidine, mannitol, sodium chloride, potassium chloride, magnesium chloride hexahydrate, calcium chloride dihydrate, and tryptophan are 4.1926 g / L, 31.0321 g / L, 6.0723 g / L, 0.974 g / L, 0.7456 g / L, 0.9036 g / L, 0.0024 g / L, and 0.4539 g / L. S3. Deep deoxygenation treatment: The first solution is subjected to deep deoxygenation treatment to reduce the dissolved oxygen content. The specific process is to introduce nitrogen gas into the first solution. The gas conditions are: gas pressure 0.5 MPa, gas flow rate 0.5 times the volume of the first solution per minute, and cumulative gas volume 5 times the volume of the first solution. S4. Isolation Filling: Conducted under a nitrogen protective atmosphere, maintained by continuous purging of a nitrogen gas curtain at a pressure of 0.1 MPa. The second solution and the first solution, which has undergone deep deoxygenation treatment, are filled in equal volumes at a volume ratio of 9:1 into the two chambers of 100 double-chamber flexible bag packaging containers that have undergone deoxygenation treatment (vacuum nitrogen filling). (The solutions are isolated between the chambers via a weak weld, and the two chambers remain isolated from filling until use.) After filling, heat-sealing clamps instantly melt and seal the bag opening. The sealed double-chamber bag products are sterilized at 115°C for 30 minutes. After sterilization, they are cooled, dried, leak-tested, and labeled to obtain the finished product. Finally, the dissolved oxygen of the first solution is tested: 1.6 ppm.

[0031] Example 7 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of a solution of α-ketoglutaric acid monopotassium salt with a concentration of 1.842 g / L and sodium chloride with a concentration of 8.766 g / L to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentrations of L-histidine hydrochloride monohydrate, L-histidine, mannitol, potassium chloride, magnesium chloride hexahydrate, calcium chloride dihydrate, and tryptophan are 4.1926g / L, 31.0321g / L, 6.0723g / L, 0.7456g / L, 0.9036g / L, 0.0024g / L, and 0.4539g / L. S3. Deep deoxygenation treatment: The first solution is subjected to deep deoxygenation treatment to reduce the dissolved oxygen content. The specific process is to introduce nitrogen gas into the first solution. The gas conditions are: gas pressure 0.5 MPa, gas flow rate 0.5 times the volume of the first solution per minute, and cumulative gas volume 5 times the volume of the first solution. S4. Isolation Filling: Under a nitrogen protective atmosphere maintained by continuous purging of a nitrogen gas curtain at a pressure of 0.3 MPa, the first solution is filled in equal volumes into 100 deoxygenated (vacuum-purified nitrogen-filled) glass bottles, and the second solution is filled in equal volumes into 100 flexible bags. The sealed products are sterilized at 115°C for 30 minutes. After sterilization, the products are cooled, dried, leak-tested, and labeled to obtain the finished product. The final dissolved oxygen level of the first solution is measured at 2.2 ppm. For use, simply mix the glass bottles containing the first solution and the flexible bags containing the second solution thoroughly.

[0032] Example 8 A method for preparing a cardioplegic solution containing α-ketoglutarate, comprising the following steps: S1. Preparation of the first solution: Prepare 10 L of a solution containing 1.842 g / L of α-ketoglutarate monopotassium salt, 54.651 g / L of mannitol, 6.710 g / L of potassium chloride, 8.132 g / L of magnesium chloride hexahydrate, 0.022 g / L of calcium chloride dihydrate, 4.085 g / L of tryptophan, and 8.766 g / L of sodium chloride to obtain the first solution; S2. Preparation of the second solution: Prepare 90L of the second solution, wherein the concentration of L-histidine hydrochloride monohydrate in the second solution is 4.1926g / L and the concentration of L-histidine is 31.0321g / L; S3. Deep deoxygenation treatment: The first solution is subjected to deep deoxygenation treatment to reduce the dissolved oxygen content. The specific process is to introduce nitrogen gas into the first solution. The gas conditions are: gas pressure 0.5 MPa, gas flow rate 0.5 times the volume of the first solution per minute, and cumulative gas volume 5 times the volume of the first solution. S4. Isolation Filling: Conducted under a nitrogen protective atmosphere, maintained by continuous purging of a nitrogen gas curtain at a pressure of 0.3 MPa. The first solution is filled in equal volumes into 100 deoxygenated (vacuum-purified nitrogen-filled) soft bags, and the second solution is filled in equal volumes into 100 soft bags. The sealed products are sterilized at 115°C for 30 minutes. After sterilization, the products are cooled, dried, leak-tested, and labeled to obtain the finished product. The final dissolved oxygen level of the first solution is measured at 2.2 ppm. For use, simply mix the first and second solutions in the soft bags thoroughly before use.

[0033] Comparative Example 1 Prepare 6L of α-ketoglutarate-containing myocardial arrest solution according to the substances and concentrations listed in Table 1. Distribute this solution evenly into 6 deoxygenated (vacuum-filled nitrogen-treated) single-chamber soft bags. Seal the product and sterilize by moist heat at 115°C for 30 minutes.

[0034] Test Example 1 Three solutions were prepared according to the concentrations of substances in Table 1, containing α-ketoglutarate monopotassium salt and L-histidine hydrochloride monohydrate, α-ketoglutarate monopotassium salt and L-histidine, and other components other than histidine compounds (L-histidine hydrochloride monohydrate and L-histidine), respectively. These solutions were numbered as sample 1, sample 2, and sample 3, and the specific components are shown in Table 2. The specific steps are as follows: Add the corresponding components required for 1L of drug solution to 800mL of purified water according to the concentration requirements in Table 1 and the design scheme in Table 2, stir until dissolved, and make up to 1L. Purge the solution with nitrogen gas at a pressure of 0.5 MPa, a flow rate of 0.5L / min, and a purging time of 10min. Obtain 3 samples. Take 500mL of each sample to determine the content of α-ketoglutarate monopotassium salt. Transfer the remaining 500mL of each sample to a three-layer co-extrusion rinsing solution, seal it in a bag, and sterilize it with moist heat at 115℃ for 30min. Cool it to room temperature under light-protected conditions and determine the content of α-ketoglutarate monopotassium salt. The results are shown in Table 2.

[0035] Table 2. Content of α-ketoglutarate monopotassium salt under different conditions Note: The concentrations of each substance in Table 2 are prepared according to the required concentrations of the corresponding substances in Table 1.

[0036] As shown in Table 2, the α-ketoglutarate monopotassium salt content was normal in all samples before sterilization. After sterilization, the α-ketoglutarate monopotassium salt content decreased in samples 1 and 2. These two samples contained L-histidine hydrochloride monohydrate and L-histidine, respectively. According to the Maillard reaction principle, compounds with carbonyl groups can undergo carbonyl-amine condensation with amino acids. α-ketoglutarate monopotassium salt contains a highly reactive α-ketocarbonyl group. The nitrogen atoms on the α-amino group of histidine and the imidazole group of the side chain can both participate in the reaction. To avoid this reaction, α-ketoglutarate monopotassium salt needs to be stored separately.

[0037] Test Example 2 The myocardial arrest fluid products prepared in Examples 1 and 6, as well as the 6 bags of myocardial arrest fluid products obtained in Comparative Example 1, were stored in a light-protected environment at 30°C. At 0 days, 3 months, 6 months, 9 months, 12 months, and 24 months, the weakly welded part of the double-chamber bags of Examples 1 and 6 was opened to connect the two chambers, and the mixture was mixed for 5 minutes to obtain a homogeneous solution. The content of α-ketoglutarate monopotassium salt in the myocardial arrest fluid products prepared in Examples 1 and 6 and the one obtained in Comparative Example 1 was measured, and the results are shown in Table 3.

[0038] Table 3. Relationship between the concentration of α-ketoglutarate monopotassium salt and time in different myocardial arrest fluid products. As can be seen from the results in Table 3, the concentration of α-ketoglutarate monopotassium salt in the myocardial stoppage solution product prepared in Example 6 of this application remains basically unchanged, which can significantly solve the stability problem of α-ketoglutarate and extend the shelf life of the myocardial stoppage solution from 12 months to 24 months. The myocardial stoppage solution product prepared in Example 1 of this application avoids degradation reaction between the two from the source by separating the solution containing α-ketoglutarate and the basic solution containing histidine compounds.

[0039] The above embodiments are only used to explain the technical solutions of this application and are not intended to limit it. Although the above embodiments have provided specific descriptions of this application, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the specific implementation methods of this application. Any modifications and equivalent substitutions that do not depart from the spirit and scope of this application should be covered within the protection scope of this application.

Claims

1. A method for preparing a cardioplegic solution containing α-ketoglutarate, characterized in that, Includes the following steps: S1. Preparation of the first solution: Dissolve the component containing α-ketoglutarate in water for injection; or dissolve the component containing α-ketoglutarate and a pharmaceutically acceptable solute other than histidine compounds in water for injection to form the first solution; S2. Preparation of the second solution: Dissolve the histidine compound in water for injection; Alternatively, a second solution may be formed by dissolving a histidine compound and a pharmaceutically acceptable solute, excluding the α-ketoglutarate component, in water for injection. S3, Separate filling: The second solution and the first solution are respectively filled into different chambers of a multi-chamber packaging container, wherein the different chambers are configured to keep the solutions separate after filling and before use; or the second solution and the first solution are respectively filled into different containers.

2. The method for preparing a myocardial arrest solution containing α-ketoglutarate as described in claim 1, characterized in that, Before the isolation filling in step S3, the first solution is further subjected to a deep deoxygenation treatment to reduce the dissolved oxygen content.

3. The method for preparing a myocardial arrest solution containing α-ketoglutarate as described in claim 1, characterized in that, In step S1, the concentration of α-ketoglutarate in the first solution is 5 to 15 times the target concentration after mixing the first solution and the second solution; the α-ketoglutarate is selected from α-ketoglutarate and / or α-ketoglutarate salt; the histidine compound is selected from L-histidine and / or L-histidine hydrochloride monohydrate.

4. The method for preparing a myocardial arrest solution containing α-ketoglutarate as described in claim 2, characterized in that, By subjecting the first solution to the aforementioned deep deoxygenation treatment, the dissolved oxygen content of the first solution is reduced to ≤3.0 ppm.

5. The method for preparing a myocardial arrest solution containing α-ketoglutarate as described in claim 4, characterized in that, The deep deoxygenation treatment includes introducing an inert gas into the first solution. The gas pressure is 0.1-0.5 MPa, the gas flow rate is 0.05-0.5 times the volume of the first solution per minute, and the cumulative gas volume is 1-5 times the volume of the first solution.

6. The method for preparing a myocardial arrest solution containing α-ketoglutarate as described in claim 1, characterized in that, In step S3, the isolation filling is carried out under an inert gas protective atmosphere, which is maintained by continuously purging an inert gas curtain at a pressure of 0.1-0.3 MPa.

7. The method for preparing a myocardial arrest solution containing α-ketoglutarate as described in claim 5 and / or 6, characterized in that, The inert gas is selected from at least one of nitrogen, argon and helium.

8. A myocardial arrest solution product, characterized in that, It is prepared by the method of any one of claims 1 to 7 for preparing myocardial arrest fluid containing α-ketoglutarate.

9. The myocardial arrest solution product according to claim 8, characterized in that, In the final sealed packaging state, the dissolved oxygen content of the first solution is ≤3.0 ppm.

10. The use of a method for preparing a myocardial arrest solution containing α-ketoglutarate as described in any one of claims 1 to 7 in the preparation of a myocardial protection arrest solution for extending shelf life and / or enabling storage and transportation, especially at ambient temperature.

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