Clevidipine butyrate concentrated solution and preparation method thereof

Abstract

The invention relates to the technical field of new drugs, in particular to a clevidipine butyrate concentrated solution and a preparation method thereof, and the composition contains clevidipine butyrate, a solubilizer, a cosolvent, a pH regulator and water for injection. Injection oil is abandoned, so that the requirements of lipid metabolism defect patients are met; no phospholipid is used in the prescription, safety risks such as hemolysis, immunoreaction and long-term toxicity and stability problems such as oxidative degradation are avoided, after phospholipid is removed, the clevidipine butyrate preparation can tolerate excessive hot-press sterilization, the sterility level guarantee degree is higher, the state of the clevidipine butyrate preparation is converted into a true solution system from a micelle system, and the stability of the clevidipine butyrate preparation is improved. The solubilizer and the cosolvent are combined, so that the solubility of clevidipine butyrate is greatly improved, dichloromethane, chloroform and other organic solvents with large toxic and side effects are not needed, the toxicity of the medicine is reduced, the stimulation to blood vessels is small, and the adverse reaction of the medicine is reduced; and the preparation method is simple and easy to amplify, and production amplification is facilitated by selecting a production process of a common injection.

Description

Technical Field

[0001] This invention relates to the field of new drug technology, specifically to a clovidil butyrate concentrate and its preparation method. Background Technology

[0002] Clavidipine butyrate is a mixture of optical isomers, its chemical name is methyl(1-butyryloxy)methyl 4-(2,3-dichlorophenyl)-1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylate, and its molecular formula is: C 21 H 23 Cl2NO6, molecular weight 456.32, structural formula as follows: Clavidipine butyrate is a dimethyl pyridine dicarboxylate, a white to off-white crystalline powder with a melting point of 138°C. It is soluble in ethanol, chloroform, ethyl acetate, and dimethyl sulfoxide, but insoluble in water. Two polymorphic forms have been identified, with form A accounting for >95%.

[0003] Clavidipine butyrate, a dihydropyridine derivative, is an ultra-short-acting intravenous calcium channel blocker used for short-term control of hypertension during the perioperative period. It exhibits high selectivity for blood vessels and myocardium, is rapidly metabolized into inactive substances in the body, and possesses strong pulse-reducing activity. It also has a vasodilatory effect on systemic blood vessels and pulmonary ducts. Clavidipine butyrate has a rapid onset and elimination of action, allowing for precise blood pressure control through dose escalation. Unlike many currently available intravenously administered antihypertensive drugs that are metabolized by the kidneys and / or liver, it is metabolized in the blood and tissues and does not accumulate in the body. It features parenteral administration, rapid onset of action, and ease of rapid adjustment.

[0004] Clavidipine butyrate is a water-insoluble drug with a solubility of only 0.0001 mg / mL in water, even lower than that of existing dihydropyridine drugs, making the preparation of injectable formulations, especially those suitable for intravenous injection, extremely difficult. To overcome this solubility problem, The Medicine Company (US) used large quantities of injectable soybean oil and egg yolk lecithin to create a water-in-oil (O / W) emulsion containing 0.5 mg / mL, marketed as Cleviprox. ® .

[0005] Domestic and foreign companies and research institutions have conducted extensive research on clovidipine butyrate formulations. The compound patent and formulation patent for clovidipine butyrate expired in 2014.

[0006] Patent document CN103237446A (US2011055617A) discloses a cloviddipine emulsion formulation containing an antimicrobial agent. This emulsion formulation comprises cloviddipine, an antimicrobial agent, lipids, an emulsifier, an osmotic pressure regulator, and water. Glycerin is used to adjust the osmotic pressure. Because the formulation contains a large amount of soybean oil and egg yolk lecithin, it is prone to bacterial growth. To enhance its antibacterial properties, the formulation adds the antimicrobial agent disodium ethylenediaminetetraacetate (EDTA), with a specific gravity of 0.001 to 0.01% w / v. However, the lipids used in the above technology are soybean oil, safflower seed oil, etc., and the emulsifier is egg yolk lecithin, etc., resulting in an oil-in-water emulsion with poor stability. Furthermore, the 2025 edition of the Chinese Pharmacopoeia, Part IV, General Chapter 0102, Injections, clearly states that antimicrobial agents should not be added to injections administered intravenously, intracranially, epidurally, or intraspinally. Therefore, the cloviddipine emulsion injection prepared by this patent poses a significant safety risk.

[0007] Patent CN104473870A discloses a clovidil butyrate liposome injection and its preparation method, comprising clovidil butyrate, injection oil, emulsifier, stabilizer, isotonic agent, pH adjuster, and water for injection. The injection oil is soybean oil for injection, and the isotonic agent is glycerol. The high-pressure microfluidic rapid preparation method is used, but the equipment cost is high, making large-scale production difficult.

[0008] Patent document CN113197853A discloses a clavidipine butyrate injection emulsion, its preparation method, and its application. The preparation method includes: shearing and mixing an oil phase and an aqueous phase to obtain a pre-emulsion; and then subjecting the pre-emulsion to a first homogenization and a second homogenization, followed by mixing with a pH adjuster. The oil phase includes clavidipine butyrate or its pharmaceutically acceptable salt or hydrate, an oil phase solvent, an emulsifier, and a co-emulsifier; the aqueous phase includes an osmotic pressure adjuster, a stabilizer, and water. The emulsion is unstable and prone to demulsification, leading to safety issues. Furthermore, its preparation method requires homogenization using a homogenizer, which is complex, costly, and difficult to scale up for production.

[0009] Patent CN120053372A discloses a clovidil butyrate micelle injection and its preparation method. This patent uses a soybean phospholipid / cholesterol mixed micelle system for drug loading to achieve solubilization. However, phospholipids are easily hydrolyzed to produce lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), which have strong surface activity. When the concentrations of LPC and LPE are too high, they can cause the rupture and dissolution of erythrocyte membranes and other cell membranes, leading to hemolysis. Soybean phospholipids contain two unsaturated fatty acid chains, which are easily oxidized to produce aldehydes and ketones, resulting in high peroxide values ​​and methoxyaniline values ​​in the formulation. Excessive peroxides can reduce the body's immune function, induce apoptosis, and consequently cause tissue damage and carcinogenesis. Therefore, the use of soybean phospholipids and bile salts in the formulation of patent CN120053372A poses certain safety risks. Furthermore, due to the instability of phospholipids and their susceptibility to hydrolysis and oxidation, the clovidil butyrate micelle injection of this patent requires storage at 2-8°C, which imposes stringent requirements on storage and transportation conditions, causing inconvenience. Furthermore, as a micelle system, when mixed with electrolyte solutions (0.9% sodium chloride injection, sodium lactate Ringer's solution), it appears as a pale blue opalescent liquid with a significantly increased particle size, posing a safety risk and limiting the use of the mixed solutions, thus restricting its clinical application.

[0010] Patent CN114796110A discloses an ethanol-free concentrate of a poorly soluble drug and a micellar solution prepared therefrom. This patent uses a composite emulsifier (phospholipid and non-phospholipid emulsifiers) and a solubilizer to achieve solubilization of the poorly soluble drug. However, this invention does not mention aseptic assurance measures, and the concentrate is quite viscous, resulting in high resistance during filtration and making scale-up production difficult. The formulation darkens in color after autoclaving, and this darkening becomes more pronounced with increasing phospholipid content. Therefore, this invention is not suitable for autoclaving, and aseptic levels are difficult to control.

[0011] As a special formulation, the size distribution of clovidil butyrate injection emulsion directly affects its pharmacokinetics and efficacy. Since the diameter of human microvessels is approximately 4-9 μm, and the diameter of pulmonary microvessels is about 5 μm, the presence of a large number of emulsion particles larger than 5 μm can lead to pulmonary embolism, alveolar tissue damage, and even death. Existing patent literature has addressed the issue of large emulsion particles, but the preparation requires multiple, even up to eight, high-pressure homogenizers connected in series for homogenization. A single homogenizer can cost millions, making the overall equipment cost prohibitively high. Secondly, excessive amounts of co-emulsifiers (oleic acid) and antibacterial agents (disodium edetate) are used in the formulation to ensure that the insoluble microparticles meet standards. However, oleic acid is easily oxidized to produce free fatty acids or other oxides, which can be harmful to the body. Simultaneously, excessive use of disodium edetate can cause a complexation reaction with calcium ions in the blood, leading to decreased blood calcium levels and posing a safety risk.

[0012] Currently available clovidipine butyrate injections use soybean oil containing long-chain triglycerides (LCTs). LCTs are esterified into long-chain fatty acids in the blood. These fatty acids are insoluble in water and require protein binding to enter cells. Once inside the cell, they need to be activated in the cytoplasm before entering the mitochondria for β-oxidation. Therefore, LCTs are easily phagocytosed and deposited by reticuloendothelial cells in organs such as the liver, spleen, and lungs, thus impairing immune function. Medium-chain triglycerides (MCTs) can be used instead of LCTs to mitigate the safety risks associated with LCTs, but this can lead to abnormal central nervous system function, such as drowsiness, nausea, and fatigue.

[0013] Therefore, to address the problems in existing clovidil butyrate formulations, such as low clovidil butyrate concentration, excessive use of soybean oil and other injectable oils, large emulsion particles that could lead to embolism, excessively high oleic acid content, poor stability, phospholipid content causing darkening of the solution after sterilization, and significant particle size increase and pale blue opalescent appearance after mixing with electrolyte solutions (0.9% sodium chloride injection, lactated Ringer's solution, etc.), posing certain safety risks and limiting the clinical use of such solutions, and the fact that glucose injection is not recommended for diabetic patients requiring blood sugar control, thus limiting its use and failing to meet clinical needs, a safer, higher-concentration injectable clovidil butyrate concentrate has become an urgent technical problem to be solved in this field. Summary of the Invention

[0014] To address the problems of existing technologies, this invention provides a high-concentration clovidil butyrate concentrate for intravenous infusion and its preparation method. The composition contains clovidil butyrate, a solubilizer, a co-solvent, a pH adjuster, and water for injection. This formulation eliminates injectable oils such as soybean oil and triglycerides, meeting the needs of patients with lipid metabolism defects, such as those with pathological hyperlipidemia, lipoid nephropathy, and acute pancreatitis with hyperlipidemia. The formulation also avoids phospholipids, thus avoiding safety risks such as hemolysis, immune reactions, and long-term toxicity, as well as stability issues such as oxidative degradation. Furthermore, it tolerates excessive autoclaving, ensuring a higher level of sterility and improving the problem of color darkening after autoclaving. Additionally, the clovidil butyrate concentrate prepared by this method is phosphate-free. The invention utilizes a true solution system, rather than a micelle system, which avoids the problems associated with micelle systems where the particle size significantly increases after mixing with electrolyte solutions, changing the appearance from a clear, transparent solution to a pale blue, opalescent liquid, and posing safety risks and limiting the use of compatible solutions. The clavidipine butyrate concentrate prepared in this invention is a true solution system without using any nanocarriers. Even after mixing with electrolyte solutions, it remains a clear liquid, thus overcoming the limitations of micelle injection solutions. The combination of solubilizers and co-solvents greatly improves the solubility of clavidipine butyrate. The formulation process of the clavidipine butyrate concentrate of this invention does not require the use of highly toxic organic solvents such as dichloromethane and chloroform, reducing drug toxicity, minimizing vascular irritation, and reducing adverse drug reactions. Furthermore, the preparation method is simple and easy to scale up, using a common injection production process, which facilitates large-scale production.

[0015] In this field, phospholipids and bile salts are often used to form a micelle system with clovidin butyrate to promote the dissolution of poorly soluble drugs. If the system does not contain phospholipids and bile salts, it cannot produce a solubilizing effect. In this invention, phospholipids and bile salts are discarded and a specific combination of solubilizers and cosolvents is used to enable it to dissolve at a higher concentration in a true solution system. This also improves the problems of increased particle size and pale blue opalescence when mixed with electrolyte solutions.

[0016] The present invention first provides a clovidipine butyrate concentrate, which mainly comprises clovidipine butyrate, solubilizer, cosolvent, pH adjuster and water for injection.

[0017] The clovidin butyrate concentrate, based on a total weight of 100%, contains 0.1%-20% clovidin butyrate, 5%-40% solubilizer, 5%-35% cosolvent, and a pH adjuster to adjust the pH of the concentrate to 6.0-8.0. The remainder is water for injection.

[0018] Preferably, the total weight of the concentrate is calculated as 100% by weight, wherein clovidin butyrate is 0.4%-16%.

[0019] The concentration of clovidin butyrate is available in concentrations of 1-200 mg / mL, with 1-10 mg / mL being commonly used, and a pH of 6.0-8.0.

[0020] The clovidin butyrate concentrate is phospholipid-free and can withstand excessive autoclaving (121°C, 15 min). No significant changes were observed in color or other test results before and after sterilization. Furthermore, the clovidin butyrate concentrate is a true solution system, compatible with electrolyte solutions, and remains a clear liquid after mixing. Since it is not a micelle system, there is no significant change in particle size. No significant changes were observed in any of the test indicators, and the use of the mixed solutions is not restricted, thus meeting the needs of different clinical patients.

[0021] The clovidin butyrate concentrate contains one or more of the following solubilizers: polyethylene glycol 15-hydroxystearate (HS 15), poloxamer, polyoxyethylene castor oil, polysorbate, and vitamin E polyethylene glycol 1000 succinate (TPGS). The co-solvent is one or more of polyethylene glycol (PEG, including PEG300 and PEG400), propylene glycol, glycerol, and anhydrous ethanol.

[0022] In some embodiments of the present invention, polyethylene glycol is selected from PEG300 and / or PEG400.

[0023] The pH adjuster is selected from at least one of acidic and alkaline pH adjusters, wherein the acidic pH adjuster includes one or more of citric acid, acetic acid, hydrochloric acid, and phosphoric acid, and the alkaline pH adjuster is sodium hydroxide and / or sodium carbonate.

[0024] The preparation method of the clovidin butyrate concentrate specifically includes the following steps: (1) Preparation of the drug solution: Weigh out clovidin butyrate, solubilizer and cosolvent according to the prescription, add them to the same container, heat and stir in a water bath at 30-80℃ until the solution is clear and transparent, cool to room temperature, and adjust the pH to 6.0-8.0 with pH adjuster; (2) Volume or weight determination: The drug solution obtained in step (1) is volume or weight determined, filtered, and then filled with protective gas. The filtration is carried out using a 0.22 μm filter membrane. (3) Sealing and sterilization: After dispensing, the headspace is filled with protective gas and then sealed. The product is then heat-sealed at 121°C for 15 min.

[0025] The protective gas used in the preparation method is any one of nitrogen, helium, carbon dioxide and argon. The time for introducing the protective gas is 0.5-2 h, the residual dissolved oxygen range is 0-5 mg / L, and the residual oxygen in the headspace is controlled at 0-5%.

[0026] Optionally, in step (1), after cooling to room temperature, add water for injection to 70-90% of the total preparation volume. If the amount of clovidin butyrate, solubilizer, and cosolvent is large (e.g., it exceeds 90% of the total preparation volume), the aforementioned treatment may not be performed. In step (1), after adding clovidin butyrate, solubilizer, and cosolvent, add 10-60% of the total preparation volume of water for injection. If the amount of clovidin butyrate, solubilizer, and cosolvent is large (e.g., it exceeds 60% of the total preparation volume), the above treatment can be omitted. The obtained clovidipine butyrate concentrate, as a dihydropyridine calcium channel blocker, is suitable for preparing drugs for the treatment of hypertension when oral antihypertensive drugs are not applicable or cannot achieve satisfactory efficacy.

[0027] In specific applications, the clovidipine butyrate concentrate provided by the present invention is a high-concentration concentrate that can be diluted with aqueous solvents. Aqueous solvents that can be used for dilution include, but are not limited to, water for injection, sodium chloride injection (mass concentration 0.9%), glucose injection (mass concentration 5%), glucose (mass concentration 5%) and sodium chloride (mass concentration 0.9%) injection, 10% amino acid injection, and sodium lactate Ringer's injection. As one of the preferred embodiments, glucose injection (mass concentration 5%) or sodium chloride injection (mass concentration 0.9%) is preferred.

[0028] This invention achieves good solubility and stability through the combined use of solubilizers and cosolvents. After adding cosolvents, the stability of the drug solution is significantly improved, and it can withstand autoclaving with no significant changes in various indicators before and after sterilization. The use of solubilizers improves solubility and significantly improves the compatibility stability of the formulation. The stability of the formulation and compatibility can only be guaranteed when solubilizers and cosolvents are present at the same time, and the two have a synergistic effect. This invention does not use phospholipids or soybean oil, is a true solution system, has no color change before and after sterilization, and remains a clear and transparent system after being mixed with electrolyte solutions, with good safety and compatibility.

[0029] The concentrate prepared by this invention is a concentrated solution and needs to be diluted before clinical use. The minimum dilution ratio is 10 times. Therefore, the cosolvent accounts for a maximum of only 3.5 wt% during clinical use. The low proportion of cosolvent during clinical use will not cause pain or irritation at the injection site.

[0030] Compared with the prior art, the present invention has achieved the following beneficial effects: (1) Improve medication safety ① Marketed emulsion injections (original injection Cleviprex) ®Other literature and patents describe fat emulsion injections that use oil-phase solvents and add phospholipids, bile acids, etc. as emulsifiers or stabilizers, EDTA as an antibacterial agent, and vitamin E as an antioxidant. This invention does not contain such components (phospholipids, soybean oil, EDTA). This invention avoids the use of large amounts of soybean oil as a solvent, thereby avoiding the increase in blood lipid levels in patients caused by injection infusion, which could lead to cardiovascular harm in patients with hyperlipidemia. This invention meets the medication needs of patients with lipid metabolism defects and significantly improves patient medication safety.

[0031] ② It contains no phospholipids or bile salts, avoiding the safety issues of hemolysis, tissue damage, and carcinogenicity caused by the oxidation or hydrolysis of phospholipids, and avoiding the safety issues of hemolysis caused by bile salts in patients with severe cirrhosis, fulminant hepatic failure, or complete biliary obstruction, thus improving the safety of clinical medication. Furthermore, the phospholipid / bile salt mixed micelle system, when mixed with electrolyte solutions (0.9% sodium chloride injection, lactated Ringer's solution, etc.), exhibits a significant increase in particle size and changes in appearance from a clear liquid to a pale blue opalescent liquid, posing safety risks that limit the use of such solutions in clinical practice. The clavidipine butyrate concentrate invented in this patent is a true solution system free of phospholipids and bile salts. It was unexpectedly discovered that the product's color did not change before and after sterilization, and it is compatible with electrolyte solutions.

[0032] (2) The clovidipine butyrate concentrate of the present invention exists in the form of a true solution, which avoids large milk particles, reduces the risk of medication, and does not have the sustained-release effect that micelle systems may bring. Once the blood pressure drops to the normal range, the infusion can be stopped immediately, which is safer.

[0033] The clovidin butyrate concentrate prepared by this invention can withstand autoclaving and becomes a clear solution after sterilization. This avoids phenomena such as emulsion particle rupture, droplet aggregation, increased particle size, and localized demulsification in commercially available emulsions, and mitigates adverse reactions caused by large emulsion particles. The main advantages are as follows: a) To avoid the formation of granulomas in the lungs due to large particles in the injected fat emulsion entering the lungs during the injection process; b) Avoid large particles entering blood vessels, which can cause angiogranulomas, phlebitis, and thrombosis; c) Avoid large particles from entering other organs of the body, such as the liver and kidneys, which can cause damage; d) The concentrate provided by this invention is clear and easy to observe for visible foreign matter and insoluble particles, which can help predict medication use in advance and reduce medication risks.

[0034] (3) Improve adaptability The clovidilide concentrate prepared by this invention can be directly diluted with sterile water for injection, 0.9% sodium chloride injection, and 5% glucose injection to obtain a dosing concentration of 0.01-1 mg / mL in clinical use. The prepared solution allows for easy control of the administration rate, eliminating the need for Y-type infusion sets and "special infusion devices with precise dose adjustment," making clinical administration more flexible and convenient, and the dosage more accurate and controllable.

[0035] (4) Improve solubility The clovidilide concentrate prepared by this invention is a high-concentration concentrate, with a concentration of clovidilide reaching over 10 mg / mL, significantly improving drug solubility compared to the marketed emulsion injection (0.5 mg / mL). In clinical administration, the total infusion volume of this formulation is equal to that of the original Cleviprex injection. ® It reduces the risk of extravasation of hypertonic drug fat emulsion injections by 1 / 2 to 1 / 20, greatly reducing the occurrence of adverse reactions such as phlebitis, and has higher clinical value.

[0036] (5) Improve stability ① Intravenous emulsions fall under the category of submicroemulsions in pharmaceutical formulation, with an average particle size mostly between 100-500 nm, making them a thermodynamically unstable system. The original injectable solution, Cleviprex, is an example. ® Once removed from the refrigerator and brought to room temperature, it cannot be returned to the refrigerator. Its shelf life at room temperature is no more than 2 months. It has poor stability and requires strict storage and transportation conditions, which can easily lead to waste during clinical use.

[0037] The clovidilin butyrate concentrate prepared by this invention can be directly formulated and used with good compatibility stability. Furthermore, the solution remained clear and showed no crystallization during the stability study. This demonstrates that the composition of this invention overcomes the disadvantage of fat emulsions not being suitable for long-term or temporary frozen storage, improving the convenience of transportation, storage, and use. The concentrated clovidilin butyrate solution prepared by this invention is a true solution that can withstand autoclaving. Terminal sterilization at 121°C for 15 minutes greatly ensures the sterility level of the injection. After sterilization, it is a clear solution with good stability, effectively avoiding phenomena such as droplet aggregation, increased particle size, and localized demulsification that occur after sterilization and long-term storage of intravenous emulsions.

[0038] ② This formulation does not contain phospholipids, thus avoiding instability caused by phospholipid hydrolysis and oxidation, and improving formulation stability. Therefore, it can be stored at room temperature, improving the convenience of transportation, storage and clinical use.

[0039] (6) The formulation process is simple and easy to scale up for production. Existing fat emulsion preparation processes require complex procedures such as high-pressure homogenization to obtain emulsion particles, and these processes are difficult and costly to control. In contrast, the system of this invention requires no special equipment, is easy to control, and can utilize a common water-based injection production route. Therefore, the process provided by this invention is simple, has low production costs, and is easy to implement. Existing patent literature uses highly toxic organic solvents such as dichloromethane and chloroform, which do not meet green environmental protection requirements and hinder scale-up production. This invention uses a small amount of solvent, thus facilitating scale-up production. Attached Figure Description

[0040] Figure 1 The particle size distribution of the injection solution prepared in Comparative Example 3 after sterilization is shown in the figure. Figure 2 The particle size distribution of the injection solution prepared in Comparative Example 3 after sterilization and recombination with 0.9% sodium chloride injection solution is shown in the figure after 0 hours. Figure 3 The image shows the appearance of the concentrated solution prepared in Example 1 after sterilization. Figure 4 The image shows the appearance of the concentrated solution prepared in Example 5 after sterilization. Figure 5 The high-performance liquid chromatogram of the concentrated solution prepared in Example 1 after sterilization is shown. Figure 6 The high-performance liquid chromatogram of the related substances determination of the concentrate prepared in Example 1 after sterilization is shown. Figure 7 The image shows the appearance of the concentrated solution prepared in Example 1 after sterilization and then mixed with 0.9% sodium chloride injection. Detailed Implementation

[0041] The following embodiments are used to illustrate the present invention, but are not intended to limit the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they should be performed in accordance with the techniques or conditions described in the literature in the field, or in accordance with the product manual.

[0042] In the embodiments, the water for injection and the drug solution after volume adjustment were both filled with protective gas. In the embodiments and comparative examples of this patent, nitrogen was used as the protective gas, and the time for introducing the protective gas was 0.5-2 h, while the dissolved oxygen was controlled within the range of 0-5 mg / L. The residual oxygen content in the headspace before sealing after dispensing was controlled within 0-5%.

[0043] Example 1: A clovidipine butyrate concentrate, the formulation of which is as follows: Clovirdipine butyrate 0.5g; HS 15 15g; PEG400 15g; Adjust the pH of the solution to 6.5 before volume adjustment using 0.5 mol / L sodium hydroxide solution or 0.1 mol / L citric acid solution; Add water for injection to a final volume of 100 mL.

[0044] The preparation method includes the following steps: (1) Preparation of drug solution: Weigh clovidipine butyrate, HS 15 and PEG400 according to the prescription, add them to the same beaker, add 50 ml of water for injection, heat and stir in a 50°C water bath until the solution is clear and transparent, cool to room temperature, add water for injection to 90% (90 mL) of the total preparation volume, and adjust the pH to 6.5 using 0.5 mol / L sodium hydroxide solution or 0.1 mol / L citric acid solution; (2) Volume or weight determination: The solution obtained in step (1) is made up to volume, filtered and then filled with nitrogen. A 0.22 μm filter membrane is used for filtration. (3) Sealing and sterilization: Dispense into 5 mL: 25 mg containers, seal with protective gas in the headspace, and autoclave at 121°C for 15 min.

[0045] The sterilized sample is a colorless, clear liquid, as shown in the appearance. Figure 3 The pH value is 6.5.

[0046] Example 2: A clovidin butyrate concentrate, the formulation of which is as follows: Clovidipine butyrate 2.5 g; HS 15 100 g; Glycerol 40 g; Adjust the pH of the solution to 6.5 before volume adjustment using 0.5 mol / L sodium carbonate solution or 0.1 mol / L citric acid solution; Add water for injection to a final volume of 500 mL.

[0047] The preparation method includes the following steps: (1) Preparation of the drug solution: Weigh out clovidin butyrate, polyethylene glycol 15-hydroxystearate solubilizer and glycerol cosolvent according to the prescription, add them to the same beaker, add 300ml of water for injection, heat and stir in a 50℃ water bath until the solution is clear and transparent, cool to room temperature, add water for injection to 450ml, and adjust the pH to 6.5 using 0.5mol / L sodium carbonate solution or 0.1mol / L citric acid solution; (2) Volume or weight determination: The solution obtained in step (1) is made up to volume, filtered and then filled with nitrogen. A 0.22 μm filter membrane is used for filtration. (3) Sealing and sterilization: Dispense into 10 mL: 50 mg, seal with protective gas in the headspace, and autoclave at 121°C for 15 min.

[0048] The concentrate, after sterilization, is a clear, colorless liquid with a pH of 6.5.

[0049] Example 3 A clovidilide concentrate, the formulation of which is as follows: Clovidipine butyrate 2.5 g; Polyoxyethylene (35) castor oil 120 g; PEG 400 50 g; Adjust the pH of the solution to 6.5 before volume adjustment using 0.5 mol / L sodium carbonate solution or 0.1 mol / L citric acid solution; Add water for injection to a final volume of 500 mL.

[0050] The preparation method is the same as in Example 2.

[0051] The concentrate is available in a 10 mL:50 mg ratio. After sterilization, it is a colorless, clear liquid with a pH of 6.6.

[0052] Example 4 A clopidogrel butyrate concentrate, the formulation of which is as follows: Clovirdipine butyrate 5 g; Polysorbate 80 100 g; 60 g of anhydrous ethanol; Adjust the pH of the solution to 6.4 before volume adjustment using 0.5 mol / L sodium hydroxide solution or 0.1 mol / L acetic acid solution; Add water for injection to a final volume of 500 mL.

[0053] The preparation method includes the following steps: (1) Preparation of drug solution: Weigh chlorvidipine butyrate, polysorbate 80 and anhydrous ethanol according to the prescription, add them to the same beaker, add 280ml of water for injection, heat and stir in a 50℃ water bath until the solution is clear and transparent, cool to room temperature, add water for injection to 90% (450mL) of the total preparation volume, and adjust the pH to 6.4 using 0.5mol / L sodium hydroxide solution or 0.1mol / L acetic acid solution; (2) Volume or weight determination: The solution obtained in step (1) is made up to volume, filtered and then filled with nitrogen. A 0.22 μm filter membrane is used for filtration. (3) Sealing and sterilization: Dispense into 5mL:50mg containers, seal with protective gas in the headspace, and autoclave at 121℃ for 15 min.

[0054] The concentrate is available in a 5 mL: 50 mg ratio. After sterilization, it is a colorless, clear liquid with a pH of 6.4.

[0055] Example 5: A clovidin butyrate concentrate, the formulation of which is as follows: Clovirdipine butyrate 20 g; HS 15 40 g; 35 g of anhydrous ethanol; Adjust the pH of the solution to 6.5 before volume adjustment using 0.1 mol / L sodium hydroxide solution or 0.1 mol / L phosphoric acid solution; Add water for injection to a final volume of 100 mL.

[0056] The preparation method includes the following steps: (1) Preparation of the drug solution: Weigh out chlorvidipine butyrate, 15-hydroxystearic acid polyethylene glycol solubilizer and anhydrous ethanol co-solvent according to the prescription, add them to the same beaker, heat and stir in a water bath at 75°C until the solution is clear and transparent, cool to room temperature, and adjust the pH to 6.5 with 0.1 mol / L sodium hydroxide solution or 0.1 mol / L phosphoric acid solution; (2) Volume or weight determination: The solution obtained in step (1) is made up to volume, filtered and then filled with nitrogen. A 0.22 μm filter membrane is used for filtration. (3) Sealing and sterilization: Dispense into 1 mL: 200 mg, seal with protective gas in the headspace, and autoclave at 121°C for 15 min.

[0057] The sterilized sample is a colorless, clear liquid, as shown in the appearance. Figure 4 The pH value is 6.5.

[0058] Example 6 A clovidilide concentrate, the formulation of which is as follows: Clovirdipine butyrate 10 g; TPGS 35 g; 10 g of glycerol; 20 g of anhydrous ethanol; Adjust the pH of the solution to 6.5 before volume adjustment using 0.1 mol / L sodium carbonate solution or 0.01 mol / L hydrochloric acid solution; Add water for injection to a final volume of 100 mL.

[0059] The preparation method includes the following steps: (1) Preparation of the drug solution: Weigh out clovidin butyrate, TPGS solubilizer, glycerol cosolvent, and anhydrous ethanol cosolvent according to the prescription, add them to the same beaker, add 10 ml of water for injection, heat and stir in a 70°C water bath until the solution is clear and transparent, cool to room temperature, add water for injection to 90 mL, and adjust the pH to 6.5 using 0.1 mol / L sodium carbonate solution or 0.01 mol / L hydrochloric acid solution; (2) Volume or weight determination: The solution obtained in step (1) is made up to volume, filtered and then filled with nitrogen. A 0.22 μm filter membrane is used for filtration. (3) Sealing and sterilization: Dispense into 1 mL: 100 mg, seal with protective gas in the headspace, and autoclave at 121°C for 15 min.

[0060] The sterilized sample was a colorless, clear liquid with a pH of 6.5.

[0061] Comparative Example 1 The clopidogrel injection solution was prepared according to the formulation disclosed in patent CN114796110A, and the formulation is as follows: Clovirdipine butyrate 1 g (1%); HS 15 45 g (45%); Egg yolk lecithin 5 g (5%); Propylene glycol 49g (49%).

[0062] The preparation method includes the following steps: (1) Weigh the prescribed amount of clovidipine butyrate and egg yolk lecithin into a 250ml beaker, add the prescribed amount of propylene glycol, stir in a 60℃ water bath for 1 min, then add the prescribed amount of HS 15, stir in a 60℃ water bath until clear, and obtain a concentrated solution. (2) Sealing and sterilization: The above concentrated liquid is dispensed, and after being filled with protective gas in the headspace, it is sealed and sterilized by autoclaving at 121°C for 15 min.

[0063] Comparative Example 2 Clavidipine injection was prepared according to the formula disclosed in patent CN114796110A, as follows: Clovirdipine butyrate 1 g (1%); HS 15 45 g (45%); Egg yolk lecithin 10 g (10%); Propylene glycol 44 g (44%).

[0064] The preparation method is the same as that of Comparative Example 1.

[0065] Results of Comparative Examples 1 and 2: The color of the samples darkened after sterilization, as shown in Table 1. The color of the samples failed the color test after sterilization, and the color of the sample in Comparative Example 2 was darker than that in Comparative Example 1. This indicates that the problem of darkening of the color after sterilization becomes more serious with increasing phospholipid concentration. In contrast, all examples produced clear solutions before and after sterilization, and the formulation containing phospholipids is not resistant to autoclaving. Therefore, the concentrated solution disclosed in this patent, due to its phospholipid content, is not resistant to autoclaving, and its sterility level is difficult to guarantee.

[0066] Table 1. Color inspection of Comparative Examples 1-2 before and after sterilization (according to the solution color inspection method of Chinese Pharmacopoeia 2025 Edition, Part IV, 0901) .

[0067] Comparative Example 3 The clavidipine butyrate micelle injection solution was prepared according to the formulation disclosed in Example 1 of patent CN120053372A, and the formulation is as follows: Clovidipine butyrate 500mg; Soybean lecithin 5.33g; Glycinecholic acid 6.67g; Sodium hydroxide 0.57g; Add water for injection to a final volume of 100 ml.

[0068] The preparation method includes the following steps: (1) Weigh out the prescribed amounts of clovidin butyrate, soybean lecithin, and glycocholic acid, add 5 ml of methanol and 2 ml of anhydrous ethanol to dissolve completely, and rotary evaporate at 40°C for 4 hours to obtain a loose film. (2) Add the prescribed amount of sodium hydroxide, add 80 ml of nitrogen-filled water for injection, stir to dissolve, transfer to the loose film obtained in step (1), stir at 300 rpm and 40°C water bath for 30 min to make it completely hydrated, and adjust the pH to 6.5 with 1 mol / L sodium hydroxide solution and 0.5 mol / L citric acid solution; (3) The solution obtained in (2) was brought to a final volume of 100 ml, filtered through a 0.22 μm filter membrane, and then purged with nitrogen. (4) Fill the ampoule with nitrogen (20s / ampoule), dispense (5ml / ampoule), seal, and autoclave at 121℃ for 15 min.

[0069] The particle size of the sterilized sample was measured to be 2.714 nm. The results are shown below. Figure 1 .

[0070] Comparative Example 4 Referring to Example 1 of patent CN120053372A, a formulation for preparing clovidin butyrate micelle injection was disclosed. A co-solvent and solubilizer were added to the formulation to prepare the sample. The formulation is as follows: Clovidipine butyrate 500mg; Soybean lecithin 5.33g; Glycinecholic acid 6.67g; Sodium hydroxide 0.57g; HS 15 15g; PEG 400 15g; Add water for injection to a final volume of 100 ml.

[0071] The preparation method includes the following steps: (1) Weigh out the prescribed amounts of clovidin butyrate, soybean lecithin, and glycocholic acid, add 5 ml of methanol and 2 ml of anhydrous ethanol to dissolve completely, and rotary evaporate at 40°C for 4 hours to obtain a loose film. (2) Add the prescribed amount of sodium hydroxide, HS 15, PEG 400 and 40 ml of nitrogen-filled water for injection, stir to dissolve, transfer to the loose film obtained in step (1), stir at 300 rpm and 40°C in a water bath for 30 min to ensure complete hydration, and adjust the pH to 6.5 with 1 mol / L sodium hydroxide solution and 0.5 mol / L citric acid solution; (3) The solution obtained in (2) was brought to a final volume of 100 ml, filtered through a 0.22 μm filter membrane, and then purged with nitrogen. (4) Fill the ampoule with nitrogen (20s / ampoule), dispense (5ml / ampoule), seal, and autoclave at 121℃ for 15 min.

[0072] The particle size of the sterilized sample was measured to be 2.868 nm. When mixed with 0.9% sodium chloride injection, it showed a pale blue opalescent color with a particle size of 51.28 nm.

[0073] The sterilized preparations of Comparative Example 3 and Comparative Example 4 were mixed with 0.9% sodium chloride injection: 5 ml of the sterilized drug solution was diluted with 0.9% sodium chloride injection to 50 ml. The diluted solutions were both pale blue opalescent liquids. The particle size of the Comparative Example 3 sample after mixing with 0.9% sodium chloride injection was 50.75 nm. (See attached image) Figure 2 Compared with the original particle size of 2.714 nm, the particle size of the sample was significantly increased. The particle size of the sample in Comparative Example 4 after being mixed with 0.9% sodium chloride injection was 51.28 nm, which was significantly increased compared with the original particle size of 2.868 nm. The appearance of the samples changed from clear liquid to light blue opalescent liquid, which poses a safety risk for clinical application.

[0074] The comparative sample exhibited the same problem when mixed with other electrolyte solutions such as sodium lactate Ringer's solution: a significant increase in particle size and a change in appearance from a clear liquid to a pale blue opalescent color. This micellar injection solution showed a significant increase in particle size when mixed with electrolyte solutions. Even with the addition of solubilizers and co-solvents in the formulation, the problem of significantly increased particle size and a pale blue opalescent appearance persisted, posing a safety risk for clinical use. Therefore, as long as the micellar system exists, its compatibility with electrolyte solutions is poor. This patent, by removing phospholipids, creatively discovered that the solution can be mixed not only with glucose injection but also with electrolyte injection, without limiting the types of solutions used, and can withstand autoclaving, ensuring a higher level of sterility.

[0075] Experimental Example 1: Investigation of the Type and Amount of Co-solvent The effects of the type and amount of cosolvent added on the injection were investigated in the formulation of fixed Example 1, including clavidipine butyrate (API) and the type of solubilizer (polyethylene glycol 15-hydroxystearate, hereinafter referred to as HS 15). As shown in Table 2, (F7 did not add water before the water bath, but added water to 90% of the total preparation volume after cooling to room temperature, and then adjusted the pH to make up to volume or weight; F8 did not add water before the water bath and after cooling to room temperature, but only adjusted the pH to make up to volume or weight).

[0076] Table 2 Summary of Cosolvent Testing Formulations Following the formulation and preparation method of Example 1 above, the type and amount of cosolvent were adjusted to prepare nine batches of concentrated solution. After autoclaving at 121°C for 15 minutes, the solutions were observed using a clarity meter (illuminance: 4000 Lx). It was observed that the F0 sample showed significant precipitation at the bottom, while the remaining solutions were clear liquids without stratification or precipitation. This demonstrates that the addition of the cosolvent helps improve the solubility of clovidin butyrate and enhances the stability of the concentrated solution.

[0077] Experimental Example 2: Investigation of Solubilizer Types The effects of changing the type and amount of solubilizer on the injection were investigated using the fixed formulation of clovidin butyrate (API) and solubilizer (PEG 400) in Example 1, as shown in Table 3. (For F16, no water was added before the water bath, but water was added to 90% of the total preparation volume after cooling to room temperature, and the pH was adjusted before making up to volume or weight. For F17, no water was added before the water bath and after cooling to room temperature, and the pH was adjusted before making up to volume or weight.)

[0078] Table 3 Summary of Solubilizer Formulas According to the prescription and preparation method of Example 1 above, the type and amount of solubilizer were adjusted to prepare 9 batches of concentrated solution. After autoclaving at 121°C for 15 minutes, no precipitation or stratification was observed in any of the 9 batches of concentrated solution, and all of them could withstand autoclaving.

[0079] The compatibility stability of the nine batches of concentrated solutions was further investigated. Formulations F9-F17 were diluted 10-fold and 20-fold with 5% glucose injection, respectively, and the stability of the solutions was observed after 24 hours at room temperature. After 4 hours, obvious precipitation was observed at the bottom of the F9 solution under a clarity tester (illuminance: 4000 Lx). The other formulations showed good stability within 24 hours, with no precipitation and no significant changes in appearance.

[0080] It is evident that adding a solubilizer to a cosolvent can improve the compatibility and stability of the formulation.

[0081] Experimental Example 3: Methods for determining drug loading, content, and related substances The determination of content and related substances shall be carried out according to the following methods.

[0082] 1. Content determination method Chromatographic conditions: Octadecylsilane-bonded silica gel (Sunniest C18, specification: 250×4.6mm, 5 μm) was used as the stationary phase; mobile phase A was methanol-acetonitrile-10 mM NaH2PO4 (approximately 1.2 g of anhydrous sodium dihydrogen phosphate was dissolved in water and diluted to 1000 mL, and the pH was adjusted to 3.0 with phosphoric acid) (volume ratio 25:25:50), and mobile phase B was acetonitrile-water (volume ratio 90:10) (volume ratio of mobile phase A to mobile phase B was 20:80); the detection wavelength was 239 nm; and the injection volume was 10 μL.

[0083] Reference solution: Take about 25 mg of clovidin butyrate reference standard, place it in a 50 mL volumetric flask, add methanol and shake to dissolve and dilute to the mark, shake well; accurately measure 2 mL, place it in a 20 mL volumetric flask, dilute to the mark with solvent (methanol-acetonitrile-water, volume ratio 40:40:20), shake well.

[0084] Test solution: Accurately measure 2 mL of the test solution under the related substances section, place it in a 20 mL volumetric flask, dilute to the mark with solvent (methanol-acetonitrile-water, volume ratio 40:40:20), and shake well.

[0085] 2. Related Substances Testing Methods Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase (Sunniest C18, specification: 250×4.6mm, 5 μm); mobile phase A was methanol-acetonitrile-10 mM NaH2PO4 (approximately 1.2 g of anhydrous sodium dihydrogen phosphate was dissolved in water and diluted to 1000 mL, and the pH was adjusted to 3.0 with phosphoric acid) (volume ratio 25:25:50), and mobile phase B was acetonitrile-water (volume ratio 90:10); the detection wavelength was 239 nm; the injection volume was 20 μL, and gradient elution was performed according to the method shown in Table 4.

[0086] Table 4 Gradient elution table for related substances test method of clovidipine butyrate concentrate (percentages in the table are volume ratios) Test solution: Accurately measure 1 mL of this product and place it in a 10 mL volumetric flask. Dilute to the mark with methanol and shake well.

[0087] The above schemes (F2-F4 in Experimental Example 1, F11-13 in Experimental Example 2, and Examples 1-6) were tested according to the above content and related substance determination methods, and the results are shown in Table 5; the content detection results of Example 1 are shown in Table 5. Figure 5The chromatograms for the detection of related substances are as follows: Figure 6 As shown in Table 5, the concentrations of the concentrated solutions prepared in each example did not change significantly before and after autoclaving.

[0088] Table 5. Detection results of content, related substances, etc. for each scheme (related substances were detected after sterilization). .

[0089] Experimental Example 4 Compatibility Stability Compositions prepared in Examples 1, 2, 3, 4, 5, 6 and Experimental Examples F9 and F11-F13 were diluted with 0.9% sodium chloride injection, 5% glucose injection, and lactated Ringer's solution to concentrations of clovidin butyrate of 0.5 mg / mL and 0.25 mg / mL, respectively. The solutions were then placed at room temperature (25°C) for 24 h to assess their stability under clinical use conditions. The results are shown in Tables 6-8. The appearance of the sample from Example 1 after reconstitution with 0.9% sodium chloride injection is shown in Table 6. Figure 7 .

[0090] Table 6. Results of Physical Stability Tests of Sample Compatibility Solutions Note: ① "-" indicates "no crystals precipitated", "+" indicates "crystals precipitated", and " / " indicates "not detected"; ②The reagent used for mixing is 5% glucose injection, and the concentration of the mixed solution is 0.5 mg / mL.

[0091] Table 7. Content determination results of the compatibility solution in Example 3 Table 8. Results of determination of related substances (total impurities) in the compatibility solution of Example 3 As shown in Table 6, precipitation was observed after the F9 sample (without solubilizer in the formulation) was placed at room temperature for 4 hours after preparation, while no precipitation was observed after the other samples containing both solubilizer and co-solvent were placed at room temperature for 24 hours. As shown in Tables 7 and 8, the dilution stability was investigated in conjunction with the content and related substance analysis methods in Experiment 3. The results showed that after the sample of Example 3 was mixed with 5% glucose injection, 0.9% sodium chloride injection, and sodium lactate Ringer's solution, and placed at room temperature for 24 hours, the content and related substance results did not change significantly compared with 0 hours, indicating good stability.

[0092] Experimental Example 5: Accelerated and Long-Term Tests The clovidil butyrate concentrates in Examples 2-6, and F5, F6, and F15 of Experiments 1 and 2, were subjected to stability studies under the following conditions. Appearance, properties, and pH changes were recorded. The clovidil butyrate content and related substances were determined according to Experiment 3 above. The detection methods for related substances (total impurities) and their content were consistent with those in Experiment 3. The results are shown in Table 9. Long-term test: 5℃±3℃; Accelerated test: 25℃±2℃ / 60%RH±5%RH.

[0093] Table 9. Results of stability studies in accelerated and long-term tests. Conclusion: As shown in the table above, the compositions prepared in each of the tested examples did not show significant changes in appearance, pH, content, related substances, and other indicators after being placed under long-term stability and accelerated stability conditions for 6 months.

[0094] The above results indicate that the formulations prepared in each example exhibit good stability. The clavidipine butyrate concentrate was designed for storage at room temperature, and based on the accelerated stability test results, its shelf life is tentatively set at 2 years.

[0095] For those skilled in the art, the specific embodiments are merely exemplary descriptions of the present invention. Obviously, the specific implementation of the present invention is not limited to the above-described manner. Any non-substantial improvements made using the inventive concept and technical solution of the present invention, or the direct application of the inventive concept and technical solution to other situations without modification, are all within the protection scope of the present invention.

Claims

1. A concentrated solution of clopidogrel butyrate, characterized in that, It contains clovidipine butyrate, solubilizer, cosolvent, pH adjuster, and water for injection; The total weight of the concentrate is calculated on a 100% weight basis, of which clovidin butyrate is 0.1%-20%, solubilizer is 5%-40%, cosolvent is 5%-35%, pH adjuster is used to adjust the pH of the concentrate to 6.0-8.0, and the remainder is water for injection; The solubilizer is one or more of the following: 15-hydroxystearic acid polyethylene glycol ester, poloxamer, polyoxyethylene castor oil, polysorbate, and vitamin E polyethylene glycol 1000 succinate. The co-solvent is one or more of polyethylene glycol, propylene glycol, glycerol, and anhydrous ethanol.

2. The clovidilic acid concentrate according to claim 1, characterized in that, The total weight of the concentrate is calculated on a 100% weight basis, of which clovidipine butyrate accounts for 0.4%-16%.

3. The clovidilide butyrate concentrate according to claim 1, characterized in that, Polyethylene glycol PEG300 and / or PEG400 are selected.

4. The clovidilic acid concentrate according to any one of claims 1-3, characterized in that, The pH adjuster is selected from at least one of acidic and alkaline pH adjusters, wherein the acidic pH adjuster includes one or more of citric acid, acetic acid, hydrochloric acid, and phosphoric acid, and the alkaline pH adjuster is sodium hydroxide and / or sodium carbonate.

5. The clovidilic acid concentrate according to any one of claims 1-3, characterized in that, The clovidin butyrate concentrate has a strength of 1-200 mg / mL and a pH of 6.0-8.

0.

6. The method for preparing the clovidilic acid concentrate according to any one of claims 1-5, characterized in that, The preparation method specifically includes the following steps: (1) Preparation of drug solution: Weigh clovidipine butyrate, solubilizer and cosolvent according to the prescription and add them to the container. Heat and stir in a water bath at 30-80℃ until the solution is clear and transparent. After cooling to room temperature, adjust the pH to 6.0-8.0 with pH adjuster. (2) Adjust the volume or weight: Adjust the volume or weight of the drug solution obtained in step (1), filter it and fill it with protective gas. The filter membrane is 0.22 μm. (3) Sealing and sterilization: After dispensing, the headspace is filled with protective gas and then sealed. The product is then heat-sealed at 121°C for 15 min.

7. The method for preparing clovidilic acid concentrate according to claim 6, characterized in that, The protective gas is any one of nitrogen, helium, carbon dioxide and argon. The protective gas is introduced for 0.5-2 hours. The dissolved oxygen residue range is 0-5 mg / L. The headspace residual oxygen content is controlled at 0-5%.

8. The method for preparing clovidilic acid concentrate according to claim 6, characterized in that, In step (1), after cooling to room temperature, add water for injection to 70-90% of the total preparation volume.

9. The method for preparing clovidilic acid concentrate according to claim 8, characterized in that, In step (1), after adding clovidin butyrate, solubilizer, and cosolvent, add 10-60% of the total preparation volume of water for injection.

10. A clovidipine butyrate injection solution, obtained by diluting the clovidipine butyrate concentrate according to any one of claims 1-5 with an aqueous solvent, wherein the aqueous solvent is selected from water for injection, sodium chloride injection, glucose injection, glucose sodium chloride injection, amino acid injection, and sodium lactate Ringer's injection.

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