Eutectic solvent injection of l-4-borophenylalanine and methods of making and using the same

Abstract

This invention relates to the field of pharmaceutical formulation technology, and particularly to a eutectic solvent injection containing L-4-boronphenylalanine, its preparation method, and its application. The eutectic solvent injection comprises L-BPA and a NADES matrix. The NADES matrix includes a hydrogen bond acceptor and a hydrogen bond donor, with a molar ratio of hydrogen bond acceptor to hydrogen bond donor of 1:(1-5). The concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200-500 mg / ml. After being diluted 10 times with 5% glucose injection, the osmotic pressure of this eutectic solvent injection is within the isotonic range of human veins, its viscosity meets clinical infusion safety standards, it exhibits excellent rheological properties, and it poses no vascular irritation or infusion risk. During storage, the solution remains clear, impurities are controllable, and the drug content remains stable. This invention solves the problem of intravenous administration of boron drugs and has promising application prospects in the field of BNCT targeted therapy.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical formulation technology, and in particular to eutectic solvent injections loaded with L-4-boronphenylalanine, their preparation methods and applications. Background Technology

[0002] Boron neutron capture therapy (BNCT) is a novel form of radiation therapy that works by delivering boron neutrons into the patient's body containing boron neutrons. 0 Boron carrier B, after selective enrichment of boron in tumor tissue, is used to irradiate the tumor site with a superthermal neutron beam.¹ 0 B captures a neutron and undergoes nuclear fission, producing alpha particles with high linear energy transfer (LET). 4 He) and recoil lithium core ( 7 Li). These heavy particles have a range of only one cell diameter (5-9 μm), enabling them to precisely kill tumor cells at the cellular scale with minimal damage to surrounding normal tissues, thus achieving targeted therapy at the cellular level.

[0003] The clinical consensus for BNCT is to use a "two-step infusion" (TSI) method: First, a rapid infusion of 200 mg / kg / h over 2 hours (cumulative 400 mg / kg) to quickly reach the plateau concentration; second, a continuous infusion of 100 mg / kg / h over 1 hour during neutron irradiation (cumulative 100 mg / kg) to maintain stable blood boron concentration and ensure a constant dose rate during irradiation. According to the standard protocol, the entire process, from drug preparation and infusion to the end of irradiation, requires at least 3 hours, which places demands on the physical stability of the diluted formulation.

[0004] L-4-boronophenylalanine (L-BPA) is currently the most widely used and mature boron carrier in clinical practice worldwide. As a tyrosine analog, L-BPA can be specifically taken up by L-amino acid transporter 1 (LAT1, SLC7A5), which is highly expressed on the surface of tumor cells, and has shown good enrichment effects in various malignant tumors (including glioblastoma, head and neck cancer, melanoma, etc.). However, the clinical application of L-BPA has long been limited by its extremely poor water solubility. The L-BPA molecule contains an amino group (-NH2), a carboxyl group (-COOH), and a boronyl group (-B(OH)2), and there are extremely strong hydrogen bonds and electrostatic interactions between the molecules. Its solubility in pure water at room temperature is only 1.2–1.6 mg / ml, which is far from meeting the needs of clinical administration. The recommended dose of L-BPA two-stage infusion for clinical BNCT treatment is 500 mg / kg body weight. Based on a body weight of 60 kg, 30 g of the drug needs to be administered. If a 1.5 mg / ml aqueous solution is used, an infusion volume of 18-25 L is required, which is almost impractical in clinical practice.

[0005] To solve the solubility problem of L-BPA, the existing technologies mainly adopt the following two strategies: (1) Complexation method, which is currently the most widely used strategy in clinical practice, such as BPA-fructose complex (BPA-F), which uses the ortho-dihydroxyl group of fructose to form a borate ester complex with the boron atom of BPA, which can increase the solubility of BPA to about 20-30 mg / ml; however, this method has obvious defects: ① It requires large volume infusion (900-1500 mL) for dispensing, such as Stella Corporation's 9000mg / 300ml packaging; ② Polysaccharide-borate ester complex is prone to oxidation and discoloration, affecting shelf life; ③ High concentrations of sugars are contraindicated for diabetic patients, such as the risk of lactic acidosis and hyperuricemia from large doses of fructose infusion; (2) pH adjustment method, which adjusts the pH to 9-10 by adding alkaline reagents such as sodium hydroxide, and uses the carboxyl group of BPA to form salt to increase solubility, but alkaline preparations are highly irritating to blood vessels, and long-term infusion can easily cause phlebitis, and the stability of BPA molecules decreases under high pH conditions, making them prone to degradation. Summary of the Invention

[0006] In view of this, the purpose of this invention is to provide an injection of L-4-boron-phenylalanine in a eutectic solvent, its preparation method and application, which achieves ultra-high concentration drug loading of L-BPA and solves the problem of poor water solubility of L-BPA.

[0007] The present invention solves the above-mentioned technical problems through the following technical means:

[0008] On one hand, embodiments of the present invention provide a eutectic solvent injection containing L-4-boronphenylalanine, comprising L-4-boronphenylalanine and a NADES matrix, wherein the NADES matrix comprises a hydrogen bond acceptor and a hydrogen bond donor, the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:(1-5), and the concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200-500 mg / ml.

[0009] In one alternative embodiment, the hydrogen bond acceptor is selected from one or a mixture of two of betaine and choline, preferably betaine; and / or, the hydrogen bond donor is selected from at least one of sorbitol, xylitol, fructose, citric acid, lactic acid, and malic acid, preferably sorbitol.

[0010] In one optional embodiment, the NADES matrix further includes water for injection, which accounts for 5 to 40% of the mass of the NADES matrix.

[0011] In one alternative embodiment, the hydrogen bond acceptor is betaine, and the hydrogen bond donor is sorbitol; the NADES matrix is ​​prepared solely from betaine and sorbitol.

[0012] In one optional embodiment, the NADES matrix is ​​prepared solely from hydrogen bond acceptors and hydrogen bond donors; the eutectic solvent injection further includes a viscosity modifier, the amount of which is 5-10% of the total mass of the eutectic solvent injection; the viscosity modifier is glycerol or propylene glycol.

[0013] In one alternative embodiment, the eutectic solvent injection further contains an additive selected from at least one of pH adjusters, isotonic adjusters, antioxidants, and metal complexing agents.

[0014] On the other hand, embodiments of the present invention provide a method for preparing a eutectic solvent injection containing L-4-boronphenylalanine, comprising the following steps: weighing a hydrogen bond acceptor and a hydrogen bond donor and adding them to a container, heating to 70-90°C and stirring at a constant temperature for 1-3 hours to obtain a NADES matrix; controlling the temperature of the NADES matrix at 30-60°C, adding L-4-boronphenylalanine, and stirring at a constant temperature for 2-5 hours to obtain a eutectic solvent injection; wherein the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:(1-5), and the concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200-500 mg / ml.

[0015] In one optional embodiment, the hydrogen bond acceptor and hydrogen bond donor are weighed and added to a container containing water for injection. The mixture is heated to 70–85°C and stirred at a constant temperature for 1–2 hours to obtain a NADES matrix. The temperature of the NADES matrix is ​​controlled at 30–40°C, and L-4-boronphenylalanine is added. The mixture is stirred at a constant temperature for 2–5 hours to obtain a eutectic solvent injection. The water for injection accounts for 5–40% of the mass of the NADES matrix.

[0016] Alternatively, weigh the hydrogen bond acceptor and hydrogen bond donor and add them to an anhydrous container. Heat to 80-90°C and stir for 2-3 hours to obtain the NADES matrix. Control the temperature of the NADES matrix at 45-60°C, add L-4-boronphenylalanine, and stir for 3-5 hours to obtain the eutectic solvent injection.

[0017] Alternatively, weigh the hydrogen bond acceptor and hydrogen bond donor and add them to an anhydrous container. Heat to 75–85°C and stir at a constant temperature for 1–2 hours to obtain a NADES matrix. Control the temperature of the NADES matrix at 35–50°C, add L-4-boronphenylalanine and a viscosity modifier, and stir at a constant temperature for 2–4 hours to obtain a eutectic solvent injection. The amount of the viscosity modifier is 5–10% of the total mass of the eutectic solvent injection, and the viscosity modifier is glycerol or propylene glycol.

[0018] In one optional embodiment, the eutectic solvent injection is sterilized by filtration through a 0.22 μm microporous membrane, aseptically dispensed into vials or ampoules, and sealed.

[0019] Alternatively, the eutectic solvent injection can be dispensed into vials or ampoules, sealed, and subjected to moist heat sterilization at 120–125°C for 10–20 minutes.

[0020] On the other hand, embodiments of the present invention provide the application of the above-mentioned eutectic solvent injection containing L-4-boronphenylalanine in the preparation of boron-loaded injections, wherein the boron-loaded injection is a boron-carrying drug for boron neutron capture therapy.

[0021] One or more technical solutions provided in the embodiments of the present invention have at least the following technical effects or advantages:

[0022] (1) The L-4-boron phenylalanine-loaded eutectic solvent injection of the present invention utilizes the large hydrogen bond network of NADES to significantly improve the solubility of L-BPA, break through the upper limit of solubility in aqueous system, achieve high concentration drug loading, and meet the high boron dosage requirement of BNCT.

[0023] (2) The L-4-boron phenylalanine-loaded eutectic solvent injection of the present invention has a hydrogen bond environment inside NADES that can stabilize the boron atom structure, inhibit the hydrolytic degradation of L-BPA, and improve the storage stability of the formulation.

[0024] (3) The L-4-boronphenylalanine-loaded eutectic solvent injection of the present invention is a high-concentration drug-loaded stock solution with a concentration of 200-500 mg / ml of L-4-boronphenylalanine. Based on 300 mg / ml, this concentration is about 230 times that of pure aqueous solution (about 1.3 mg / ml) and about 10 times that of traditional fructose complexation preparations (about 30 mg / ml). The product volume is reduced to 1 / 10 of the original, solving the industry pain point of poor water solubility of L-BPA and providing convenient drug transportation.

[0025] (4) The eutectic solvent injection containing L-4-boron phenylalanine of the present invention can be stabilized for more than 6 hours without precipitation after dilution, which fully covers the 4-6 hour time window requirement of the two-stage infusion method of BNCT, and solves the problems of discoloration and easy precipitation after dilution of existing fructose preparations, thus greatly improving the safety of clinical use.

[0026] (5) The method for preparing the L-4-boron phenylalanine-loaded eutectic solvent injection of the present invention first prepares an intact eutectic matrix, and then adds the drug according to the viscosity of the system by temperature control (30-40°C for aqueous system and 45-60°C for anhydrous system). Under the premise of ensuring complete dispersion and dissolution of the drug, the L-BPA is avoided from being thermally decomposed and the boric acid group is degraded under high temperature conditions. The reaction temperature is mild and does not require high temperature and high pressure. The requirements for industrial production equipment are low.

[0027] (6) The method for preparing the eutectic solvent injection of L-4-boron phenylalanine of the present invention reduces the rate and risk of oxidation and discoloration, and can effectively improve the shelf life. Detailed Implementation

[0028] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0029] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0030] Natural deep eutectic solvents (NADES) are a new type of green solvent that has emerged in recent years. They are eutectic mixtures formed by intermolecular hydrogen bond interactions between naturally derived hydrogen bond acceptors (HBAs) and hydrogen bond donors (HBDs). NADES are characterized by simple preparation, low vapor pressure, strong solubility, and good biocompatibility, showing great promise for drug delivery. However, current research on NADES drug delivery mainly focuses on transdermal and oral administration routes, with very little research on intravenous NADES formulations. Furthermore, in existing technologies, high-concentration NADES stock solutions cannot be directly injected intravenously due to their extremely high osmotic pressure (typically >2000 mOsm / kg) and high viscosity; dilution is necessary. However, the dilution process may disrupt the hydrogen bond network of NADES, leading to drug precipitation.

[0031] Therefore, this application proposes a eutectic solvent injection containing L-4-boronphenylalanine, comprising L-4-boronphenylalanine and a NADES matrix. The NADES matrix includes a hydrogen bond acceptor and a hydrogen bond donor, with a molar ratio of hydrogen bond acceptor to hydrogen bond donor of 1:(1-5). The concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200-500 mg / ml.

[0032] The above-described technical solution utilizes the extensive hydrogen bond network of NADES to significantly improve the solubility of L-BPA, breaking through the upper limit of solubility in aqueous systems and achieving high-concentration drug loading, thus meeting the high boron dosage requirements of BNCT. Simultaneously, the hydrogen bond environment within NADES stabilizes the boron atom structure, inhibiting the hydrolytic degradation of L-BPA and improving the storage stability of the formulation. Furthermore, the flowability can be controlled by adjusting the ratio of hydrogen bond acceptors and hydrogen bond donors, making injection administration feasible.

[0033] The concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200–500 mg / ml, preferably 300–350 mg / ml, forming a high-concentration drug-loaded stock solution. At 300 mg / ml, this concentration is approximately 230 times the solubility in pure aqueous solution (approximately 1.3 mg / ml) and approximately 10 times that of traditional fructose complexation preparations (approximately 30 mg / ml), allowing for a reduction in dispensing volume to 1 / 10 of the original.

[0034] In one embodiment, the hydrogen bond acceptor is selected from one or a mixture of two of betaine and choline; and / or, the hydrogen bond donor is selected from at least one of sorbitol, xylitol, fructose, citric acid, lactic acid, and malic acid.

[0035] The above technical solutions utilize injectable-grade, biocompatible hydrogen bond acceptors and donors that are all metabolizable in vivo, exhibiting low cytotoxicity and meeting the safety requirements for intravenous injection. The system viscosity, hydrogen bond strength, and L-BPA solubility can be flexibly adjusted to suit different process requirements. Betaine (trimethylglycine) is an endogenous natural substance widely found in beets, spinach, and other plants and animals. It is a commonly used drug in clinical practice for treating hyperhomocysteinemia and has been included in the pharmacopoeias of many countries as an excipient for injection, demonstrating excellent safety. Betaine molecules contain quaternary ammonium cations and carboxyl anions, possessing a strong hydrogen bond forming ability. As a NADES for HBA formation, it has a unique solubilizing advantage for amino acid drugs. Choline (usually choline chloride) is an essential B vitamin for the human body and an important component of cell membrane phospholipids. Its biocompatibility has been fully verified, making it suitable as an HBA component for injectable NADES. Betaine and choline can be mixed in any proportion to form a mixed HBA system that can adjust the viscosity, pH, and solubilizing ability of NADES. Sorbitol is a classic injectable excipient listed in the pharmacopoeia and is widely used in large-volume parenteral solutions such as mannitol injection and compound amino acid injection; its safety for intravenous administration has been clinically validated for decades. The sorbitol molecule contains six hydroxyl groups, which can form a dense three-dimensional hydrogen bond network with HBA, and simultaneously form multiple hydrogen bonds with the amino, carboxyl, and boronic groups of L-BPA molecules, resulting in a significant solubilizing effect. Xylitol, as a sugar metabolism intermediate, can also be safely used for intravenous administration.

[0036] In one embodiment, the NADES matrix further includes water for injection, which accounts for 5-40% of the mass of the NADES matrix, thus preparing an aqueous NADES matrix.

[0037] The addition of an appropriate amount of water for injection has multiple functions: First, as a hydrogen bond modifier, it significantly reduces the viscosity of the NADES system, improves flowability, and makes production operations such as drug addition, filtration, and dispensing at 30–40°C feasible. Second, water molecules can embed into the HBA-HBD hydrogen bond network, acting as a "lubricant" to make the hydrogen bond network more flexible rather than destroying the hydrogen bonds. Third, studies have shown that when the water content is below 40% (w / w), the supramolecular hydrogen bond structure of NADES remains intact and will not disintegrate due to the addition of water. The supramolecular structure of NADES can be maintained within a water content range of 5–40%. Considering the system viscosity, processing feasibility, and stability after dilution, the optimal water content is 5–20%. Fourth, the presence of an appropriate amount of water can significantly improve the stability of the diluted drug solution and slow down the crystallization kinetics process.

[0038] In one embodiment, the hydrogen bond acceptor is betaine and the hydrogen bond donor is sorbitol, and an anhydrous NADES matrix is ​​prepared; the NADES matrix is ​​prepared solely from betaine and sorbitol.

[0039] The NADES matrix prepared solely from betaine and sorbitol exhibits superior biocompatibility, is water-free, and completely avoids the risk of L-BPA degradation and precipitation caused by water competing for hydrogen bonds; at the same time, the anhydrous system has the best chemical stability and is suitable for long-term storage; however, its high viscosity makes processing and production difficult.

[0040] In one embodiment, the NADES matrix is ​​an anhydrous NADES matrix prepared solely from hydrogen bond acceptors and hydrogen bond donors; the eutectic solvent injection further includes a viscosity modifier, the amount of which is 5-10% of the total mass of the eutectic solvent injection, and the viscosity modifier is glycerol or propylene glycol.

[0041] Using glycerol or propylene glycol as viscosity modifiers to reduce viscosity, compared to adding water for injection, does not fiercely compete for NADES hydrogen bonds, thus preserving the original hydrophobic microenvironment and L-BPA stability to the greatest extent. Simultaneously, glycerol and propylene glycol offer high injection safety and also provide some solubilizing, moisturizing, and stabilizing effects; furthermore, adding glycerol or propylene glycol allows for controlled reduction of system viscosity, improving injection flowability, and exhibiting better sterilization tolerance compared to aqueous systems.

[0042] In one embodiment, the eutectic solvent injection further contains an adjuvant selected from at least one of a pH adjuster, an isotonic adjuster, an antioxidant, and a metal complexing agent. The pH adjuster is used to stabilize the system pH and inhibit the hydrolysis of L-BPA borate groups; the pH adjuster may be selected from hydrochloric acid solution, sodium hydroxide solution, or a citrate buffer pair. The isotonic adjuster is used to optimize intravenous infusion safety and reduce vascular irritation; the isotonic adjuster may be selected from sodium chloride solution. The antioxidant and metal complexing agent are used to reduce metal ion-catalyzed oxidative degradation of the drug; the antioxidant may be sodium bisulfite, and the metal complexing agent may be EDTA-2Na. The amounts of the adjuvants are all conventional amounts used in the field of injections.

[0043] This application also discloses a method for preparing the above-mentioned eutectic solvent injection compound loaded with L-4-boronphenylalanine, comprising the following steps:

[0044] Weigh out hydrogen bond acceptors and hydrogen bond donors and add them to a container. Heat to 70-90℃ and stir for 1-3 hours to obtain NADES matrix. Add drugs according to system type and temperature control: for aqueous systems, control the NADES matrix temperature at 30-40℃; for anhydrous systems, control the temperature at 45-60℃; for anhydrous systems containing viscosity modifiers, control the temperature at 35-50℃. Add L-4-boronphenylalanine and stir for 2-5 hours to obtain eutectic solvent injection. Dispense into vials and seal. The molar ratio of hydrogen bond acceptors to hydrogen bond donors is 1:(1-5), and the concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200-500 mg / ml.

[0045] The above technical solution first prepares a complete eutectic matrix, and then adds the drug at a low temperature, avoiding the thermal decomposition of L-BPA and the deterioration of boric acid groups under high temperature conditions; the reaction temperature is mild, no high temperature and high pressure are required, and the requirements for industrial production equipment are low.

[0046] In one embodiment, hydrogen bond acceptor and hydrogen bond donor are weighed and added to a container containing water for injection. The mixture is heated to 70-85°C and stirred at a constant temperature for 1-2 hours to obtain a NADES matrix. The temperature of the NADES matrix is ​​controlled at 30-40°C, and L-4-boronphenylalanine is added. The mixture is stirred at a constant temperature for 2-5 hours to obtain a eutectic solvent injection. The water for injection accounts for 5-40% of the mass of the NADES matrix, preferably 5-20%.

[0047] In another embodiment, hydrogen bond acceptor and hydrogen bond donor are weighed and added to an anhydrous container, heated to 80-90°C and stirred at a constant temperature for 2-3 hours to obtain NADES matrix. The temperature of NADES matrix is ​​controlled at 45-60°C, L-4-boronphenylalanine is added, and stirred at a constant temperature for 3-5 hours to obtain eutectic solvent injection.

[0048] In another embodiment, hydrogen bond acceptors and hydrogen bond donors are weighed and added to an anhydrous container, heated to 75-85°C and stirred at a constant temperature for 1-2 hours to obtain a NADES matrix. The temperature of the NADES matrix is ​​controlled at 35-50°C, and L-4-boronphenylalanine and a viscosity modifier are added. The mixture is stirred at a constant temperature for 2-4 hours to obtain a eutectic solvent injection. The amount of viscosity modifier used is 5-10% of the total mass of the eutectic solvent injection, and the viscosity modifier is glycerol or propylene glycol.

[0049] In one embodiment, the eutectic solvent injection is sterilized by filtration through a 0.22 μm microporous membrane under a temperature of 30–40°C, and then aseptically dispensed into vials or ampoules and sealed. The 0.22 μm microporous membrane, relying on a sieving and retention mechanism, traps microorganisms such as bacteria, yeasts, and molds in the solution (common pathogens are generally larger than 0.22 μm), thus achieving sterilization. L-4-boronphenylalanine contains boric acid functional groups, which are prone to hydrolysis and structural damage under high temperatures; the filtration and sterilization process is controlled at a mild temperature of 30–40°C to avoid thermal degradation of L-4-boronphenylalanine and ensure the integrity of the active pharmaceutical ingredient's chemical structure. Furthermore, prolonged high temperatures in some aqueous NADES systems and systems with added polyol modifiers can easily trigger hydrogen bond recombination and even phase separation; the cold sterilization process can maintain the original homogeneous liquid phase structure of the eutectic solvent.

[0050] In another embodiment, the eutectic solvent injection is dispensed into vials or ampoules, sealed, and subjected to moist heat sterilization at 120–125°C for 10–20 minutes. Terminal sterilization is achieved by relying on high-temperature saturated water vapor to penetrate the vials and using the high temperature to denature and inactivate microbial proteins and nucleic acids, thus reaching a sterility assurance level; this is a terminal sterilization process. The formulation is sterilized after sealing, and the system is not opened after sterilization to avoid the risk of secondary contamination. Both aqueous and anhydrous systems can use terminal moist heat sterilization. L-4-boronphenylalanine can withstand short-term high temperatures under NADES hydrogen bond protection; for aqueous systems, the stability and related substances need to be examined after terminal sterilization, or a 0.22 μm sterile filtration process can be used.

[0051] The eutectic solvent injection of this application is sealed in vials or ampoules in high-concentration stock solution and stored and transported in a cool, dark place. Before use, it is diluted to a clinically appropriate concentration of L-BPA (e.g., 20–40 mg / ml) using a commonly used intravenous infusion diluent, gently shaken, and immediately administered intravenously. The diluted solution should be infused within 6 hours. The diluent is one of 5% glucose injection, water for injection, or 0.9% sodium chloride injection, preferably 5% glucose injection. The dilution ratio is 5–20 times (v / v), preferably 8–12 times. After dilution, the concentration of NADES in the solution remains above the supramolecular structure formation concentration, effectively inhibiting L-BPA crystallization.

[0052] 5% glucose injection was chosen as the dilution medium because the stability of the NADES supramolecular hydrogen bond network depends on the hydrogen bond interaction between HBA and HBD. 0.9% sodium chloride injection contains 154 mM Na⁺ and Cl⁻ ions. The presence of these electrolyte ions may electrostatically compete with HBA (the cationic group of betaine / choline), interfering with the integrity of the NADES supramolecular clusters to some extent, accelerating the dissociation of the hydrogen bond network, and thus shortening the drug's stabilization time. In contrast, 5% glucose injection is a non-electrolyte solution, containing no ionic components. Dilution does not introduce additional ionic interference, which is more conducive to maintaining the integrity of the NADES supramolecular hydrogen bond network, ensuring that the diluted drug solution remains stable without precipitation for more than 4 hours. Furthermore, glucose is a physiological energy source for the human body, metabolically safe, and has no electrolyte load, making it suitable for a wider range of patients.

[0053] When the eutectic solvent injectable of this application is used for boron neutron capture therapy, it is administered via a two-stage infusion method: The first stage involves infusion at a rate of 200 mg / kg / h for 2 hours (cumulative administration of 400 mg / kg) to rapidly reach plateau blood concentration; the second stage involves continuous infusion at a rate of 100 mg / kg / h for 1 hour during neutron irradiation (cumulative administration of 100 mg / kg) to maintain stable blood boron concentration. The total dose is 500 mg / kg body weight. Based on a standard body weight of 60 kg, the total L-BPA dosage is 30 g. If diluted to a clinically common concentration of 20–40 mg / ml, the total infusion volume over 3 hours is controlled within the range of 750–1500 mL, meeting the safety requirements for intravenous fluid load in clinical practice.

[0054] In specific implementation, the following examples can be referred to:

[0055] Example 1 (Betaine + Sorbitol + Water, sterile filtration)

[0056] The eutectic solvent injection compound of this embodiment includes the following components:

[0057] Betaine 117.15 g (1.0 mol), sorbitol 546.5 g (3.0 mol), water for injection 35 g (5% w / w of NADES matrix), L-4-boronphenylalanine 174.7 g.

[0058] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0059] Weigh out betaine and sorbitol according to the formula, add them to a stainless steel mixing tank equipped with stirring and temperature control, add water for injection, start stirring, heat to 85℃ and stir for 2 hours until a colorless, clear, and transparent homogeneous liquid is formed, then cool to 40℃ to obtain the betaine-sorbitol NADES matrix. Maintain the mixing tank temperature at 40℃, and slowly add L-4-boronphenylalanine to the above NADES matrix in batches (1 / 4 of the volume each time, stirring for 10 minutes before adding the next batch), continuing to stir at a constant temperature for 5 hours until the drug is completely dissolved and the solution is clear and transparent. Filter the solution under two-stage sterilization using a 0.22μm polyethersulfone microporous membrane at 30–35℃. Dispense the filtrate into 20 mL vials under Class A laminar flow, filling each vial with 10 mL, and then capping and coring to obtain the high-concentration drug-loaded stock solution.

[0060] Example 2 (Betaine + Sorbitol + Water, sterile filtration)

[0061] The eutectic solvent injection compound of this embodiment includes the following components:

[0062] Betaine 117.15 g (1.0 mol), sorbitol 546.5 g (3.0 mol), water for injection 442.5 g (40% w / w of NADES matrix), L-4-boronphenylalanine 474 g.

[0063] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0064] Weigh out betaine and sorbitol according to the formula, add them to a stainless steel mixing tank equipped with stirring and temperature control, add water for injection, start stirring, heat to 70℃ and stir for 1 hour until a colorless, clear, and transparent homogeneous liquid is formed, then cool to 30℃ to obtain the betaine-sorbitol NADES matrix. Maintain the mixing tank temperature at 30℃, and slowly add L-4-boronphenylalanine to the above NADES matrix in batches (1 / 4 of the volume each time, stirring for 10 minutes before adding the next batch), continuing to stir at a constant temperature for 2 hours until the drug is completely dissolved and the solution is clear and transparent. Filter the solution under two-stage sterilization using a 0.22μm polyethersulfone microporous membrane at 30–35℃. Dispense the filtrate into 20 mL vials under Class A laminar flow, filling each vial with 10 mL, stoppering and capping to obtain the high-concentration drug-loaded stock solution.

[0065] Example 3 (Betaine + Sorbitol + Water, sterile filtration)

[0066] Betaine 117.15 g (1.0 mol), sorbitol 546.5 g (3.0 mol), water for injection 221.2 g (25% w / w of NADES matrix), L-4-boronphenylalanine 379.2 g.

[0067] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0068] Weigh out betaine and sorbitol according to the formula, add them to a stainless steel mixing tank equipped with stirring and temperature control, add water for injection, start stirring, heat to 80℃ and stir for 1.5 hours until a colorless, clear, and transparent homogeneous liquid is formed, then cool to 35℃ to obtain the betaine-sorbitol NADES matrix. Maintain the mixing tank temperature at 35℃, and slowly add L-4-boronphenylalanine to the above NADES matrix in batches (1 / 4 of the volume each time, stirring for 10 minutes before adding the next batch), continuing to stir at a constant temperature for 3 hours until the drug is completely dissolved and the solution is clear and transparent. Filter the solution under two-stage sterilization using a 0.22μm polyethersulfone microporous membrane at 30–35℃. Dispense the filtrate into 20 mL vials under Class A laminar flow, filling each vial with 10 mL, stoppering and capping to obtain the high-concentration drug-loaded stock solution.

[0069] Example 4 (Betaine + Sorbitol + Water, terminal moist heat sterilization)

[0070] Betaine 117.15 g (1.0 mol), sorbitol 546.5 g (3.0 mol), water for injection 221.2 g (25% w / w of NADES matrix), L-4-boronphenylalanine 379.2 g.

[0071] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0072] Weigh out betaine and sorbitol according to the formula, add them to a stainless steel mixing tank equipped with stirring and temperature control, add water for injection, start stirring, heat to 80℃ and stir at a constant temperature for 1.5 hours until a colorless, clear, and transparent homogeneous liquid is formed, then cool to 35℃ to obtain the betaine-sorbitol NADES matrix. Maintain the temperature of the mixing tank at 35℃, and slowly add L-4-boronphenylalanine to the above NADES matrix in batches (1 / 4 of the volume each time, stirring for 10 minutes before adding the next batch), continue stirring at a constant temperature for 3 hours until the drug is completely dissolved and the solution is clear and transparent. Dispense into 20 mL vials, filling each vial with 10 mL, stoppering and crimping the cap, and sterilize at 121℃ for 15 minutes.

[0073] Example 5 (choline chloride + sorbitol + water, sterile filtration)

[0074] The eutectic solvent injection compound of this embodiment includes the following components:

[0075] Choline chloride 139.6 g (1 mol), sorbitol 546.5 g (3 mol), water for injection 228.7 g (25% w / w of NADES matrix), L-4-boronphenylalanine 392 g.

[0076] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 3. Finally, the solution is sterilized by two stages using a 0.22μm polyethersulfone microporous membrane under the condition of keeping warm at 30-35℃. The filtrate is dispensed into 20 mL vials under the laminar flow of stage A, with 10 mL filled into each vial. The vials are then capped and stoppered to obtain the high-concentration drug-loaded stock solution.

[0077] Example 6 (Betaine + Choline Chloride + Sorbitol + Water, sterilized filtration)

[0078] The eutectic solvent injection compound of this embodiment includes the following components:

[0079] Betaine 58.6g (0.5mol), choline chloride 69.8g (0.5mol), sorbitol 546.5g (3mol), water for injection 225g (25% w / w of NADES matrix), L-4-boronphenylalanine 385.7g.

[0080] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 3. Finally, the solution is sterilized by two stages using a 0.22μm polyethersulfone microporous membrane under the condition of keeping warm at 30-35℃. The filtrate is dispensed into 20 mL vials under the laminar flow of stage A, with 10 mL filled into each vial. The vials are then capped and stoppered to obtain the high-concentration drug-loaded stock solution.

[0081] Example 7 (Betaine + Xylitol + Water, sterilized filtration)

[0082] The eutectic solvent injection compound of this embodiment includes the following components:

[0083] Betaine 117.15 g (1 mol), xylitol 456.4 g (3 mol), water for injection 191.2 g (25% w / w of NADES matrix), L-4-boronphenylalanine 327.7 g.

[0084] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Embodiment 3. Finally, the solution is sterilized by two stages using a 0.22μm polyethersulfone microporous membrane under the condition of keeping warm at 30-35℃. The filtrate is dispensed into 20 mL vials under the laminar flow of stage A, with 10 mL filled into each vial. The vials are then capped and stoppered to obtain the high-concentration drug-loaded stock solution.

[0085] Example 8 (Betaine + Sorbitol + Water, sterile filtration)

[0086] The eutectic solvent injection compound of this embodiment includes the following components:

[0087] Betaine 117.15 g (1 mol), sorbitol 182.2 g (1 mol), water for injection 99.8 g (25% w / w of NADES matrix), L-4-boronphenylalanine 171 g.

[0088] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Embodiment 3. Finally, the solution is sterilized by two stages using a 0.22μm polyethersulfone microporous membrane under the condition of keeping warm at 30-35℃. The filtrate is dispensed into 20 mL vials under the laminar flow of stage A, with 10 mL filled into each vial. The vials are then capped and stoppered to obtain the high-concentration drug-loaded stock solution.

[0089] Example 9 (Betaine + Sorbitol + Water, sterile filtration)

[0090] The eutectic solvent injection compound of this embodiment includes the following components:

[0091] Betaine 117.15 g (1 mol), sorbitol 911 g (5 mol), water for injection 342.7 g (25% w / w of NADES matrix), L-4-boronphenylalanine 587.5 g.

[0092] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Embodiment 3. Finally, the solution is sterilized by two stages using a 0.22μm polyethersulfone microporous membrane under the condition of keeping warm at 30-35℃. The filtrate is dispensed into 20 mL vials under the laminar flow of stage A, with 10 mL filled into each vial. The vials are then capped and stoppered to obtain the high-concentration drug-loaded stock solution.

[0093] Example 10 (Betaine + Sorbitol + Water + Additives, Sterilization Filtration)

[0094] The eutectic solvent injection compound of this embodiment includes the following components:

[0095] Betaine 117.15 g (1.0 mol), sorbitol 546.5 g (3.0 mol), water for injection 221.2 g (25% w / w of NADES matrix), L-4-boronphenylalanine 379.2 g.

[0096] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0097] Weigh out betaine and sorbitol according to the formula, add them to a stainless steel mixing tank equipped with stirring and temperature control, add water for injection, start stirring, heat to 80℃ and stir at a constant temperature for 1.5 hours until a colorless, clear, and transparent homogeneous liquid is formed, then cool to 35℃ to obtain the betaine-sorbitol NADES matrix. Maintain the mixing tank temperature at 35℃, slowly add L-4-boronphenylalanine to the above NADES matrix in batches (1 / 4 of the volume each time, stirring for 10 minutes before adding the next batch), then add 0.13g of injection-grade anhydrous sodium sulfite and continue stirring at a constant temperature for 3 hours until the drug is completely dissolved and the solution is clear and transparent. Finally, add 0.1mol / L hydrochloric acid solution to adjust the pH of the system to approximately 7.0. The solution was sterilized by two-stage filtration using a 0.22μm polyethersulfone microporous membrane under a temperature of 30–35℃. The filtrate was dispensed into 20 mL vials under a laminar flow condition (stage A), with 10 mL filled into each vial. The vials were then capped and sealed to obtain a high-concentration drug-loaded stock solution.

[0098] Example 11 (Betaine + Sorbitol, terminal moist heat sterilization)

[0099] The eutectic solvent injection compound of this embodiment includes the following components:

[0100] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), L-4-boronphenylalanine 284.4 g.

[0101] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0102] Weigh out betaine and sorbitol according to the formula, add them to an anhydrous stainless steel mixing tank equipped with stirring and temperature control, start stirring, heat to 80℃ and stir at a constant temperature for 2 hours, then cool to 45℃ to obtain a viscous transparent liquid, namely the betaine-sorbitol NADES matrix. Control the temperature of the mixing tank at 45℃, slowly add L-4-boronphenylalanine to the above NADES matrix (1 / 3 of the volume each time, stirring for 10 minutes before adding the next batch), continue stirring at a constant temperature for 3 hours, and dispense the resulting homogeneous solution into 20 mL vials, filling each vial with 10 mL, stoppering and crimping the caps, and sterilizing at 125℃ for 10 minutes.

[0103] Example 12 (Betaine + Sorbitol, terminal moist heat sterilization)

[0104] The eutectic solvent injection compound of this embodiment includes the following components:

[0105] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), L-4-boronphenylalanine 284.4 g.

[0106] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0107] Weigh out betaine and sorbitol according to the formula, add them to an anhydrous stainless steel mixing tank equipped with stirring and temperature control, start stirring, heat to 90℃ and stir at a constant temperature for 3 hours, then cool to 60℃ to obtain a viscous transparent liquid, namely the betaine-sorbitol NADES matrix. Control the temperature of the mixing tank at 60℃, slowly add L-4-boronphenylalanine to the above NADES matrix (1 / 3 of the volume each time, stirring for 10 minutes before adding the next batch), continue stirring at a constant temperature for 5 hours, and dispense the resulting homogeneous solution into 20 mL vials, filling each vial with 10 mL, stoppering and crimping the caps, and sterilizing at 120℃ for 20 minutes.

[0108] Example 13 (Betaine + Sorbitol, terminal moist heat sterilization)

[0109] The eutectic solvent injection compound of this embodiment includes the following components:

[0110] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), L-4-boronphenylalanine 284.4 g.

[0111] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0112] Weigh out betaine and sorbitol according to the formula, add them to an anhydrous stainless steel mixing tank equipped with stirring and temperature control, start stirring, heat to 85℃ and stir at a constant temperature for 2.5h, then cool to 50℃ to obtain a viscous transparent liquid, namely the betaine-sorbitol NADES matrix. Control the temperature of the mixing tank at 50℃, slowly add L-4-boronphenylalanine to the above NADES matrix (1 / 3 of the volume each time, stirring for 10min before adding the next batch), continue stirring at a constant temperature for 4h, and dispense the resulting homogeneous solution into 20 mL vials, filling each vial with 10mL, stoppering and crimping the caps, and sterilizing at 121℃ for 15min.

[0113] Example 14 (Betaine + Sorbitol + Glycerol, terminal moist heat sterilization)

[0114] The eutectic solvent injection compound of this embodiment includes the following components:

[0115] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), glycerol 49.9 g, L-4-boronphenylalanine 284.4 g.

[0116] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0117] Weigh out betaine and sorbitol according to the formula, add them to an anhydrous stainless steel mixing tank equipped with stirring and temperature control, start stirring, heat to 75℃ and stir at a constant temperature for 1 hour, then cool to 35℃ to obtain a viscous transparent liquid, namely the betaine-sorbitol NADES matrix. Control the temperature of the mixing tank at 35℃, slowly add L-4-boronphenylalanine (1 / 3 of the amount each time, stirring for 10 minutes before adding the next batch) and glycerol to the above NADES matrix, continue stirring at a constant temperature for 2 hours, and dispense the resulting homogeneous solution into 20mL vials, filling each vial with 10mL, stoppering and crimping the caps, and sterilizing at 121℃ for 15 minutes.

[0118] Example 15 (Betaine + Sorbitol + Glycerol, terminal moist heat sterilization)

[0119] The eutectic solvent injection compound of this embodiment includes the following components:

[0120] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), glycerol 121.0 g, L-4-boronphenylalanine 427.0 g.

[0121] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0122] Weigh out betaine and sorbitol according to the formula, add them to an anhydrous stainless steel mixing tank equipped with stirring and temperature control, start stirring, heat to 85℃ and stir at a constant temperature for 2 hours, then cool to 50℃ to obtain a viscous transparent liquid, namely the betaine-sorbitol NADES matrix. Control the temperature of the mixing tank at 50℃, slowly add L-4-boronphenylalanine (1 / 3 of the amount each time, stirring for 10 minutes before adding the next batch) and glycerol to the above NADES matrix, continue stirring at a constant temperature for 4 hours, and dispense the resulting homogeneous solution into 20mL vials, filling each vial with 10mL, stoppering and crimping the caps, and sterilizing at 121℃ for 15 minutes.

[0123] Example 16 (Betaine + Sorbitol + Glycerol, terminal moist heat sterilization)

[0124] The eutectic solvent injection compound of this embodiment includes the following components:

[0125] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), glycerol 71.0 g, L-4-boronphenylalanine 375.0 g.

[0126] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0127] Weigh out betaine and sorbitol according to the formula, add them to an anhydrous stainless steel mixing tank equipped with stirring and temperature control, start stirring, heat to 80℃ and stir at a constant temperature for 1.5h, then cool to 40℃ to obtain a viscous transparent liquid, namely the betaine-sorbitol NADES matrix. Control the temperature of the mixing tank at 40℃, slowly add L-4-boronphenylalanine (1 / 3 of the amount each time, stirring for 10min before adding the next batch) and glycerol to the above NADES matrix, continue stirring at a constant temperature for 3h, and dispense the resulting homogeneous solution into 20 mL vials, filling each vial with 10mL, stoppering and crimping the caps, and sterilizing at 121℃ for 15min.

[0128] Example 17 (Betaine + Sorbitol + Propylene Glycol, Terminal Moist Heat Sterilization)

[0129] The eutectic solvent injection compound of this embodiment includes the following components:

[0130] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), propylene glycol 74.9 g, L-4-boronphenylalanine 197.9 g.

[0131] The preparation method of the eutectic solvent injection material in this embodiment is as follows:

[0132] Weigh out betaine and sorbitol according to the formula, add them to an anhydrous stainless steel mixing tank equipped with stirring and temperature control, start stirring, heat to 80℃ and stir at a constant temperature for 1.5h, then cool to 40℃ to obtain a viscous transparent liquid, namely the betaine-sorbitol NADES matrix. Control the temperature of the mixing tank at 40℃, slowly add L-4-boronphenylalanine (1 / 3 of the amount each time, stirring for 10min before adding the next batch) and propylene glycol to the above NADES matrix, continue stirring at a constant temperature for 3h, and dispense the resulting homogeneous solution into 20 mL vials, filling each vial with 10mL, stoppering and crimping the caps, and sterilizing at 121℃ for 15min.

[0133] Example 18

[0134] The eutectic solvent injection compound of this embodiment includes the following components:

[0135] Betaine 117.15 g (1 mol), sorbitol 182.2 g (1 mol), glycerol 51.7 g, L-4-boronphenylalanine 273 g.

[0136] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0137] Example 19

[0138] The eutectic solvent injection compound of this embodiment includes the following components:

[0139] Betaine 117.15 g (1 mol), sorbitol 364.3 g (2 mol), glycerol 63.8 g, L-4-boronphenylalanine 329 g.

[0140] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0141] Example 20

[0142] The eutectic solvent injection compound of this embodiment includes the following components:

[0143] Betaine 117.15 g (1 mol), sorbitol 728.7 g (4 mol), glycerol 78.2 g, L-4-boronphenylalanine 402 g.

[0144] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0145] Example 21

[0146] The eutectic solvent injection compound of this embodiment includes the following components:

[0147] Betaine 117.15 g (1 mol), sorbitol 910.9 g (5 mol), glycerol 85.2 g, L-4-boronphenylalanine 438 g.

[0148] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0149] Example 22

[0150] The eutectic solvent injection compound of this embodiment includes the following components:

[0151] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), glycerol 54.0 g, L-4-boronphenylalanine 385 g.

[0152] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0153] Example 23

[0154] The eutectic solvent injection compound of this embodiment includes the following components:

[0155] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), glycerol 62.5 g, L-4-boronphenylalanine 380 g.

[0156] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0157] Example 24

[0158] The eutectic solvent injection compound of this embodiment includes the following components:

[0159] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), glycerol 71.0 g, L-4-boronphenylalanine 375 g.

[0160] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0161] Example 25

[0162] The eutectic solvent injection compound of this embodiment includes the following components:

[0163] Betaine 117.15 g (1 mol), sorbitol 546.5 g (3 mol), glycerol 79.2 g, L-4-boronphenylalanine 370 g.

[0164] The preparation method of the eutectic solvent injection in this embodiment is the same as that in Example 16. Finally, it is dispensed into 20 mL vials, 10 mL per vial, capped and corked, and placed in 121°C for moist heat sterilization for 15 min.

[0165] Comparative Example 1

[0166] The injectable substance in this comparative example is a traditional BPA-fructose complex solution, in which the fructose:BPA molar ratio is 2:1.

[0167] Performance testing:

[0168] (1) Solubility test

[0169] 10 g each of purified water, traditional BPA-fructose complex solution (fructose:BPA molar ratio 2:1), and NADES matrix prepared in Examples 1-25 were taken, and excess L-4-boronphenylalanine was added. The mixtures were magnetically stirred at 25°C for 24 h to reach dissolution equilibrium. After centrifugation at 12000 rpm for 15 min, the supernatant was appropriately diluted with the mobile phase. The BPA peak area was determined by high-performance liquid chromatography (HPLC), and the solubility of BPA in each system was calculated using the external standard method. Chromatographic conditions: Agilent ZORBAX SB-C18 column (4.6 × 250 mm, 5 μm); mobile phase: methanol-water (20:80, containing 0.1% trifluoroacetic acid); flow rate: 1.0 mL / min; detection wavelength: 280 nm; column temperature: 30°C; injection volume: 20 μL. The solubility test results are shown in Table 1.

[0170] Testing revealed that the solubility of L-BPA in pure water was 1.3 mg / ml, and the solubility in the BPA-fructose complex solution was 28.5 mg / ml. The NADES matrix of Examples 1-25 of this invention exhibited a saturated solubility of 210.5–455.0 mg / g for L-4-boronphenylalanine. This demonstrates that the NADES matrix system of this invention has a highly significant solubilizing effect on L-BPA, achieving ultra-high concentration drug loading. Traditional aqueous solutions have extremely low solubility for L-BPA, making it difficult to prepare high-concentration injectable solutions. However, the NADES system in these examples relies on hydrogen bond network supramolecular interactions to significantly improve the saturated solubility of the drug. The actual drug loading in all examples was lower than the saturated solubility at 25°C, and there was no risk of crystallization during the storage of the finished product.

[0171] (2) Physical stability test

[0172] The drug administration scenario of two-stage infusion in clinical BNCT was simulated to examine the stability of high-concentration stock solution at different time points after dilution, and to verify whether it met the 4-hour baseline requirement and the 6-hour clinical time window requirement.

[0173] The drug administration scenario of two-stage infusion in clinical BNCT was simulated to examine the stability of high-concentration stock solution at different time points after dilution, and to verify whether it met the 4-hour baseline requirement and the 6-hour clinical time window requirement.

[0174] Specifically, the high-concentration drug-loaded stock solutions prepared in Examples 3, 13, and 16 were diluted with 5% glucose injection under laminar flow hood to a concentration of approximately 30 mg / ml for L-BPA. The diluted solutions were placed at room temperature of 25°C, and samples were taken at 0, 1, 2, 3, 4, 5, 6, and 8 hours after dilution for the following tests: (1) Visual inspection: The clarity of the solution was visually inspected against a black and white background to check for any visible foreign matter or crystals.

[0175] Test Results: The high-concentration drug-loaded stock solutions prepared in Examples 1, 7, and 8, after dilution with 5% glucose injection, were colorless, clear, and transparent from 0 to 6 hours. The content retention rate at 6 hours was greater than 99.3%, and the number of particles ≥10 μm was 7.8 particles / mL, 7.5 particles / mL, and 7.6 particles / mL, respectively, meeting pharmacopoeia standards. A very slight opalescence appeared at 8 hours. In contrast, the 0.9% sodium chloride injection dilution group showed slight opalescence at 5 hours and trace white spots at 6 hours, exhibiting slightly inferior stability compared to the glucose dilution group, confirming that electrolyte ions have a certain interference effect on the supramolecular structure of NADES. The BPA-fructose control preparation showed very slight opalescence at 2 hours, trace white spots at 4 hours, a small amount of crystallization at 5 hours, and a large amount of crystallization at 6 hours. DLS measurements showed that the NADES preparations (diluted with glucose) prepared in Examples 3, 13, and 16 contained only supramolecular clusters with a particle size of about 1-3 nm in the solution within 6 hours after dilution, with no aggregates or microcrystals larger than 100 nm; while the BPA-fructose control group showed nanocrystals of 100-500 nm after 2 hours, and rapidly grew into micron-sized crystals after 4 hours.

[0176] The above data results indicate that: ① The eutectic solvent injection of the present invention remains completely clear within 6 hours after dilution with 5% glucose injection, with a drug content retention rate >99%, and the number of insoluble particles meets the injection standards of the Chinese Pharmacopoeia, fully satisfying the 4-hour baseline requirement and the 6-hour clinical medication time window requirement of the BNCT two-stage infusion method; ② The stability of 5% glucose injection as a dilution medium is better than that of 0.9% sodium chloride injection, because the latter contains electrolyte ions, which may interfere with the NADES supramolecular hydrogen bond network, so 5% glucose injection is preferred; ③ Existing clinically used BPA-fructose preparations show opalescence after 2 hours of dilution, white spots are visible after 4 hours, and crystals precipitate after 5 hours, posing a high risk for clinical use and must be strictly limited to infusion within 2 hours; ④ The dilution stability of the preparation of the present invention is significantly better than that of existing fructose preparations, providing a sufficient safety window for clinical medication. This advantage stems from the continuous inhibitory effect of the NADES supramolecular hydrogen bond network on L-BPA crystallization.

[0177] (3) Osmotic pressure test

[0178] The osmotic pressure of the low-co-solubility solvent injection stock solutions prepared in Examples 1-25, after a 10-fold dilution (diluted with 5% glucose injection), and the osmotic pressure of the 5% glucose injection under the same conditions were measured using a freezing point osmometer. The stock solutions prepared in each example of this invention are high-concentration stock solutions and are not for direct intravenous administration. The test results are shown in Table 1. The osmotic pressure of the 5% glucose injection is 290 ± 5 mOsm / kg.

[0179] Test results show that the eutectic solvent injection of the present invention, after being diluted 10 times with 5% glucose injection, yields an osmotic pressure that meets the requirements for intravenous administration. In the NADES matrix of the aqueous system, the addition of water dilutes ionic solutes such as betaine, reducing the concentration of osmotic active particles in the system; increasing the proportion of sorbitol (hydrogen bond donor) further decreases the osmotic pressure. Sorbitol is a nonionic polyol that does not dissociate to generate osmotic particles, thus buffering the system's osmotic pressure. Compared to the anhydrous glycerol-free group with an average osmotic pressure of 304.7 mOsm / kg, the anhydrous glycerol-containing group had an osmotic pressure of 288-300 mOsm / kg. Glycerol, being a nonionic cosolvent, reduces the relative proportion of betaine, making the diluted solution's osmotic pressure closer to the physiological isotonic midpoint.

[0180] (4) Viscosity test

[0181] Using the eutectic solvent injections prepared in Examples 1-25 as samples, the dynamic viscosity of the stock solution at 25°C was measured using a rotational viscometer. The dynamic viscosity of the diluted solutions obtained by diluting the stock solution 10 times with 5% glucose injection at 25°C was also measured. The dynamic viscosity of the 5% glucose injection at 25°C was also measured. The test results are shown in Table 1. The dynamic viscosity of the 5% glucose injection at 25°C was 1.02 ± 0.03 cP.

[0182] Test results show that after 10-fold dilution with 5% glucose injection, the dynamic viscosity of all diluted solutions falls within the range of 1.28-1.61 cP, which is within the suitable viscosity range for intravenous infusion and meets the rheological requirements for intravenous administration. In the aqueous NADES matrix, the viscosity of the stock solution decreases significantly with increasing water content. Water molecules can break the hydrogen bond cross-linking network inside NADES, weakening molecular entanglement and resulting in a very significant viscosity reduction effect. The viscosity of the diluted solution decreases synchronously with the viscosity of the stock solution. In the aqueous formulation, water plays a dominant role in plasticizing and diluting; in the anhydrous formulation, the absence of water molecules breaking hydrogen bonds leads to a large number of hydrogen bonds associated between high-content sorbitol polyol molecules, causing an increase in viscosity. Glycerol and propylene glycol, as small-molecule polyols, insert into the gaps in the NADES hydrogen bond network, weakening molecular chain entanglement and acting as molecular lubricants.

[0183] (5) Rabbit ear vein irritation test

[0184] Twelve healthy New Zealand white rabbits were randomly divided into four groups of three each: the stock solution group of the eutectic solvent injection prepared in Example 16, the diluted drug solution group (the eutectic solvent injection prepared in Example 16 was diluted 10 times with 5% glucose), the 5% glucose injection control group, and the BPA-fructose positive control group. Administered via an indwelling needle in the marginal ear vein, the dosage was 5 mL / kg, once daily for three consecutive days.

[0185] The experimental results showed that: in the undiluted solution group, obvious redness and swelling of the ear margin appeared immediately after injection, and pathological sections showed endothelial cell edema; in the diluted solution group, no abnormalities such as redness, swelling, or congestion were observed at the injection site, and pathological sections showed intact endothelial cell morphology, with no difference compared to the 5% glucose injection control group; the BPA-fructose group showed mild redness and swelling, and pathological sections showed mild endothelial swelling. Conclusion: Direct intravenous injection of high-concentration NADES undiluted solution poses significant vascular irritation and must be diluted before use; the solution diluted 10 times has no vascular irritation and good safety.

[0186] (6) Hemolytic test

[0187] Fresh blood was collected from healthy rabbits, and a 2% erythrocyte suspension was prepared. Seven test tubes were used, and the following solutions were added respectively: a eutectic solvent injection prepared in Example 16 diluted 3 times with 5% glucose; a eutectic solvent injection prepared in Example 16 diluted 5 times with 5% glucose; a eutectic solvent injection prepared in Example 16 diluted 8 times with 5% glucose; a eutectic solvent injection prepared in Example 16 diluted 10 times with 5% glucose; a eutectic solvent injection prepared in Example 10 diluted 15 times with 5% glucose; 5% glucose injection solution; and distilled water. The tubes were incubated in a constant temperature water bath at 37°C for 4 hours. The experimental results showed that no hemolysis or erythrocyte agglutination occurred within 4 hours in any of the test tubes, with hemolysis rates all <5%, indicating no hemolysis or erythrocyte agglutination, meeting the requirements for the hemolysis test of the injection.

[0188] (7) Storage stability test

[0189] The eutectic solvent injection stock solutions prepared in Examples 1-25 were filled into 20 mL neutral borosilicate glass vials and sealed. They were placed in accelerated stability test chambers (30℃±2℃, relative humidity 65%±5%) and long-term stability test chambers (25℃±2℃, relative humidity 60%±5%) for testing, with an additional test group under cool storage conditions (20℃±2℃). Samples were taken at 0, 3, and 6 months for accelerated stability testing; samples were taken at 0, 6, and 12 months for long-term stability testing. The appearance and clarity of the solutions were observed. The content of L-BPA and the total impurity level were determined by high-performance liquid chromatography (HPLC). The storage stability of each formulation was evaluated by changes in appearance, retention rate of active ingredients, and increase in impurities during storage. All samples remained clear throughout the testing period, without turbidity, precipitation, or discoloration. Specific content and impurity data are shown in Table 2.

[0190] Test results showed that all samples remained clear throughout the entire testing period, without turbidity, precipitation, or discoloration. Using Example 3 (sterilized filtration) as a control group for the aqueous system, and comparing it with Example 4 (terminal moist heat sterilization), it was observed that after terminal moist heat sterilization at 121℃, the hydrogen bond network within the NADES matrix rearranged, forming a more dense and stable supramolecular solvation cage structure. This structure provides better encapsulation and protection for L-4-boronphenylalanine, effectively isolating oxygen and inhibiting drug oxidative degradation. After 6 months of accelerated storage, the moist heat sterilization group showed a drug retention rate of 96.8% and a total impurity rate of 0.33%; after 12 months of long-term storage, the drug retention rate was 96.0% and the total impurity rate was 0.45%. The stability was significantly better than the sterilized filtration sample with the same formulation (Example 3, long-term 12-month retention rate 95.2%, total impurity rate 0.58%), demonstrating that terminal moist heat sterilization not only does not damage the formulation performance but also improves the storage stability of the aqueous NADES injection.

[0191] The anhydrous glycerol system formulation itself exhibits excellent thermal stability, and all formulations undergo terminal moist heat sterilization at 121°C. High-temperature treatment further optimizes the glycerol-betaine-sorbitol hydrogen bond network, resulting in a 97.5%-98.2% retention rate of the active pharmaceutical ingredient in the glycerol-containing sterilized group after 6 months, with minimal impurity growth. Example 15, a high-glycerol formulation, demonstrates the best stability. Example 10, which incorporates sodium sulfite antioxidant in addition to moist heat sterilization, shows maximum suppression of degradation impurities. In summary, the eutectic solvent matrix of this invention, adapted to terminal moist heat sterilization, offers superior sterilization assurance and storage stability compared to sterile filtration processes, demonstrating significant advantages for industrial production.

[0192] (8) Preliminary verification of pharmacodynamics in animals

[0193] Twelve nude mice bearing human nasopharyngeal carcinoma CNE-2 were randomly divided into two groups of six each. The experimental group received a tail vein injection of a diluted eutectic solvent injection solution (BPA dose 200 mg / kg, diluted with 5% glucose) as described in Example 16, while the control group received an equal dose of BPA-fructose preparation. Two hours after administration, the animals were sacrificed, and tumor tissue, blood, and muscle were collected. Boron concentration in each tissue was determined using ICP-MS.

[0194] Results: The tumor boron concentration in the experimental group was 38.7 ± 5.2 μg / g, and in the control group it was 35.2 ± 4.8 μg / g (p>0.05); the T / B ratio in the experimental group was 3.6 ± 0.5, and in the control group it was 3.4 ± 0.4 (p>0.05); the T / M ratio in the experimental group was 4.8 ± 0.7, and in the control group it was 4.5 ± 0.6 (p>0.05). These results indicate that the eutectic solvent injection of the present invention delivers boron to tumors in animals with performance comparable to existing clinical formulations. The in vivo distribution and tumor targeting of L-BPA were not affected by the dosage form change, demonstrating its potential for clinical translation.

[0195] Table 1

[0196]

[0197] Table 2

[0198]

[0199] Note: ① The data for October are the initial values ​​measured immediately after the formulation is prepared; L-BPA content is calculated according to the labeled amount, and total impurities are determined by HPLC area normalization method. ② Accelerated test conditions are 30℃±2℃, RH 65%±5%, and long-term test conditions are 25℃±2℃, RH 60%±5%.

[0200] The foregoing description includes examples from this specification. Of course, for the purposes of describing this specification, it is impossible to describe every conceivable combination of components or methods; however, those skilled in the art will understand that many other combinations and arrangements are possible. Therefore, this specification is intended to cover all such changes, modifications, and variations that fall within the spirit and scope of the appended claims. Furthermore, with regard to the use of the term "comprising" in the detailed description or claims, the term is intended to be inclusive in a manner similar to the term "including," as interpreted when "comprising" is used as a transitional word in the claims.

[0201] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A eutectic solvent injection product loaded with L-4-boronphenylalanine, characterized in that, The product comprises L-4-boronphenylalanine and a NADES matrix, wherein the NADES matrix includes a hydrogen bond acceptor and a hydrogen bond donor, the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:(1-5), and the concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200-500 mg / ml.

2. The eutectic solvent injection product loaded with L-4-boronphenylalanine according to claim 1, characterized in that, The hydrogen bond acceptor is selected from one or a mixture of two of betaine and choline; and / or, The hydrogen bond donor is selected from at least one of sorbitol, xylitol, fructose, citric acid, lactic acid, and malic acid.

3. The eutectic solvent injection product loaded with L-4-boronphenylalanine according to claim 1 or 2, characterized in that, The NADES matrix also includes water for injection, which accounts for 5 to 40% of the mass of the NADES matrix.

4. The eutectic solvent injection product loaded with L-4-boronphenylalanine according to claim 1, characterized in that, The hydrogen bond acceptor is betaine, and the hydrogen bond donor is sorbitol; the NADES matrix is ​​prepared solely from betaine and sorbitol.

5. The eutectic solvent injection product loaded with L-4-boronphenylalanine according to claim 2, characterized in that, The NADES matrix is ​​prepared solely from hydrogen bond acceptors and hydrogen bond donors; the eutectic solvent injection further includes a viscosity modifier, the amount of which is 5-10% of the total mass of the eutectic solvent injection; the viscosity modifier is glycerol or propylene glycol.

6. The eutectic solvent injection product loaded with L-4-boronphenylalanine according to claim 1, characterized in that, The eutectic solvent injection also contains an additive selected from at least one of pH adjusters, isotonic adjusters, antioxidants, and metal complexing agents.

7. A method for preparing a eutectic solvent injection product loaded with L-4-boronphenylalanine, characterized in that, Includes the following steps: Weigh out hydrogen bond acceptors and hydrogen bond donors and add them to a container. Heat the container to 70-90°C and stir for 1-3 hours to obtain NADES matrix. Control the temperature of NADES matrix at 30-60°C, add L-4-boronphenylalanine, and stir for 2-5 hours to obtain eutectic solvent injection. The molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:(1-5), and the concentration of L-4-boronphenylalanine in the eutectic solvent injection is 200-500 mg / ml.

8. The preparation method according to claim 7, characterized in that, Weigh the hydrogen bond acceptor and hydrogen bond donor and add them to a container containing water for injection. Heat the mixture to 70-85°C and stir for 1-2 hours to obtain a NADES matrix. Control the temperature of the NADES matrix at 30-40°C and add L-4-boronphenylalanine. Stir for 2-5 hours to obtain a eutectic solvent injection. The water for injection accounts for 5-40% of the mass of the NADES matrix. Alternatively, weigh the hydrogen bond acceptor and hydrogen bond donor and add them to an anhydrous container. Heat to 80-90°C and stir for 2-3 hours to obtain the NADES matrix. Control the temperature of the NADES matrix at 45-60°C, add L-4-boronphenylalanine, and stir for 3-5 hours to obtain the eutectic solvent injection. Alternatively, weigh the hydrogen bond acceptor and hydrogen bond donor and add them to an anhydrous container. Heat to 75–85°C and stir at a constant temperature for 1–2 hours to obtain a NADES matrix. Control the temperature of the NADES matrix at 35–50°C, add L-4-boronphenylalanine and a viscosity modifier, and stir at a constant temperature for 2–4 hours to obtain a eutectic solvent injection. The amount of the viscosity modifier is 5–10% of the total mass of the eutectic solvent injection, and the viscosity modifier is glycerol or propylene glycol.

9. The preparation method according to claim 8, characterized in that, The eutectic solvent injection is sterilized by filtration through a 0.22μm microporous membrane, aseptically dispensed into vials or ampoules, and sealed. Alternatively, the eutectic solvent injection can be dispensed into vials or ampoules, sealed, and subjected to moist heat sterilization at 120–125°C for 10–20 minutes.

10. The use of the eutectic solvent injectable containing L-4-boronphenylalanine according to any one of claims 1-9 in the preparation of boron-loaded injections, wherein the boron-loaded injection is a boron-carrying drug for boron neutron capture therapy.

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