CN116350586A - A kind of nimodipine micellar injection and preparation method thereof - Google Patents

Abstract

The invention belongs to the field of medicines, and provides nimodipine micelle injection and a preparation method thereof, in particular relates to high-concentration nimodipine micelle injection for intravenous drip, which consists of nimodipine serving as an active ingredient, phospholipid serving as an auxiliary material, cholic acid or salt thereof, a surface modification material, an isotonic regulator, a pH regulator and water for injection. The injection not only greatly improves the solubility of nimodipine, but also adopts a physiologically compatible adjuvant phospholipid, cholic acid and a micelle system with good safety for solubilization, and adopts an innovative film dispersion-hydration process, and ethanol is abandoned as a solubilizer in the prescription, thereby obviously improving the safety of medication. The injection can be directly diluted by injection, the compatibility solution has good stability for 24 hours, special equipment and secondary nursing of an infusion pump are not needed, and the medication safety and compliance are improved.

Description

A kind of nimodipine micellar injection and preparation method thereof

Technical Field

The invention belongs to the field of medicine and provides a nimodipine micelle injection and a preparation method thereof.

Background Art

Nimodipine is a mixture of optical isomers. Its chemical name is 2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydro-3,5-pyridinedicarboxylic acid-2-methoxyethyl-(1-methylethyl) ester. Its molecular formula is C ₂₁ H ₂₆ N ₂ O ₇ , molecular weight is 418.4, and its structural formula is as follows:

Nimodipine is a dimethyl pyridine dicarboxylate, a fine crystalline yellow substance with a melting point of 125-126°C. It is soluble in ethanol, chloroform, ethyl acetate and polyethylene glycol, but insoluble in water. Nimodipine is stable in neutral and acidic media and sensitive to alkali. It is thermally stable and non-hygroscopic, but moderately sensitive to light, especially in solution; the pH value of a 1% (w/v) solution of nimodipine is 6-6.8.

Nimodipine is a second-generation 1,4-dihydropyridine calcium antagonist with high lipophilicity and easy penetration of the blood-brain barrier. In animal experiments, nimodipine can bind to L-type Ca ²⁺ channels with high affinity and high specificity, thereby inhibiting Ca ²⁺ transmembrane influx. It is inferred that nimodipine can improve the stability and functional capacity of pathological states caused by increased Ca ²⁺ ion influx in nerve cells, such as cerebral ischemia. Because it can easily penetrate the blood-brain barrier, it has the effect of selectively dilating cerebral blood vessels and significantly reversing spasms of the basilar artery and anterior spinal artery. It is clinically used to treat hypertension, stroke, migraine, subarachnoid hemorrhage and other cerebral hemorrhage diseases. It is currently the first choice for the treatment of cerebrovascular diseases, especially for Alzheimer's disease. It has a high clinical application value. Nimodipine is the first-line treatment for ischemic brain injury.

注射液由德国拜尔公司研制，于1985年4月上市，规格为50ml：10mg。尼莫地平已有多个产品进入2018版国家基药目录(片剂、胶囊：20mg、30mg)、2021版国家医保目录(甲类：口服常释剂型，乙类：注射剂)。目前尼莫地平在国内获批生产的剂型有普通片剂、胶囊剂、注射剂及口服溶液等。尽管尼莫地平具有良好的生物学效应，但其临床性能受到其低口服生物利用度(低至10％)和低水溶性(3.86μg/mL)的限制。因此，静脉给药是尼莫地平疗效的替代途径。为了达到足够的尼莫地平浓度，上市制剂通过用约40％的溶剂混合物溶解尼莫地平，即由23.7％(v/v)乙醇和17％(v/v)聚乙二醇400组成溶剂混合物。然而这种浓度在临床上应用时有许多明显的缺点：大量乙醇对酗酒或酒精代谢受损的人以及孕妇或哺乳期妇女有害。此外高浓度的乙醇可能会在注射部位引起疼痛和刺激。由于尼莫地平必须以1～2mg/h的速度慢速滴注，否则病人无法耐受其副作用，即10mg药物所需滴注时间一般需要至少5h，使用时需与配伍溶液(0.9％氯化钠注射液、5％葡萄糖注射液等)混合滴注或用特殊的三通输液器与注射液同时滴注，混合液直接输入病人体内；静脉注射最长连续输注长达三周。同时尼莫地平在乙醇溶解，在水中不溶解，尼莫地平输液为含乙醇的非水溶性制剂，当与其他输液配伍后，均可析出结晶，含量下降，降低药物疗效，也给病人带来一定程度的危险。Original drug

The injection was developed by Bayer AG of Germany and was launched in April 1985 with specifications of 50ml: 10mg. Several products of nimodipine have entered the 2018 version of the National Essential Drug Catalog (tablets, capsules: 20mg, 30mg) and the 2021 version of the National Medical Insurance Catalog (Class A: oral regular release dosage form, Class B: injection). At present, the dosage forms of nimodipine approved for production in China include ordinary tablets, capsules, injections and oral solutions. Although nimodipine has good biological effects, its clinical performance is limited by its low oral bioavailability (as low as 10%) and low water solubility (3.86μg/mL). Therefore, intravenous administration is an alternative route for the efficacy of nimodipine. In order to achieve sufficient nimodipine concentration, the marketed preparation dissolves nimodipine with about 40% solvent mixture, that is, a solvent mixture composed of 23.7% (v/v) ethanol and 17% (v/v) polyethylene glycol 400. However, this concentration has many obvious disadvantages when used clinically: large amounts of ethanol are harmful to alcoholics or people with impaired alcohol metabolism, as well as pregnant or lactating women. In addition, high concentrations of ethanol may cause pain and irritation at the injection site. Since nimodipine must be slowly dripped at a rate of 1 to 2 mg/h, otherwise the patient cannot tolerate its side effects, that is, the drip time required for 10 mg of the drug generally requires at least 5 hours. When used, it must be mixed with a compatible solution (0.9% sodium chloride injection, 5% glucose injection, etc.) for dripping or dripped simultaneously with the injection solution using a special three-way infusion set, and the mixed solution is directly injected into the patient's body; the longest continuous infusion for intravenous injection is up to three weeks. At the same time, nimodipine dissolves in ethanol and does not dissolve in water. Nimodipine infusion is a non-water-soluble preparation containing ethanol. When it is compatible with other infusions, crystals can be precipitated, the content decreases, and the efficacy of the drug is reduced, which also brings a certain degree of danger to the patient.

Patent document CN20181146629 discloses a nimodipine injection composition and a preparation method thereof, wherein a high proportion of soybean oil, medium-chain fatty oil or a mixture of soybean oil and medium-chain fatty oil is used in the prescription. Therefore, the corresponding standards for emulsification level and technical process are improved during commercial production. In addition, the chemical properties of unsaturated double bonds in unsaturated fatty acids such as soybean oil are unstable, and impurity degradation affects drug safety. Therefore, the quality standard control of indicators such as peroxide value, anisidine value and free fatty acids in the product is required to be more stringent.

Patent document CN1732936A discloses nimodipine emulsion injection and its preparation method. An appropriate amount of benzyl alcohol is added during the preparation process to improve the solubility of nimodipine in the emulsion and the stability of the preparation, making the concentration of the nimodipine emulsion relatively high. Benzyl alcohol has disinfection, preservation and local anesthetic effects. It is used as an antibacterial agent and analgesic in injections and has a hemolytic effect. When the injection for intramuscular injection in children contains benzyl alcohol, it can cause gluteal muscle contracture. The appendix of the 2020 edition of the "Chinese Pharmacopoeia" has strict restrictions on the addition of antibacterial agents to intravenous injections under the injection item.

Patent document CN112137956A discloses a method for preparing a nimodipine sustained-release emulsion injection, in which Tween 80 is used in the prescription. Currently, intravenous injections containing Tween 80 have many clinical adverse reactions, which may cause hemolytic reactions, acute hypersensitivity reactions, peripheral neurotoxicity, inhibition of P-glycoprotein activity, intrinsic anti-tumor effects, hepatotoxicity, etc. Therefore, intravenous injections containing Tween 80 should be used with caution.

Patent document CN101485632A discloses nimodipine lipid microsphere injection and its preparation method, but the lipid microsphere preparation has complex process and high cost. The lipid microsphere package is easily damaged and leaked during long-term storage, which brings clinical risks. In addition, the particle size is large, and the risk of intravenous injection is high.

Patent document CN105796490B discloses a nimodipine injection composition containing amino acids and a preparation method thereof, wherein ethanol and polyethylene glycol are used for solubilization to prepare the injection.

Patent document CN114886850A discloses a nimodipine preparation and a method for treating a disease, wherein a surfactant polysorbate 80 and an organic solvent ethanol solubilizer are used in the prescription.

Both of the above prior arts have the following disadvantages: large amounts of ethanol are harmful to alcoholics or people with impaired alcohol metabolism, as well as pregnant or lactating women. In addition, high concentrations of ethanol may cause pain and irritation at the injection site.

Patent document CN101088503A discloses nimodipine freeze-dried powder injection and its preparation method, wherein Tween 80, polyethylene glycol 400, propylene glycol and ethanol are used in the formula. There are many clinical reports on adverse reactions of propylene glycol, which limits its clinical application.

Patent document CN102525917B discloses a nimodipine micellar injection and a method for preparing the same. Activated carbon is used in the formula, and the content is significantly reduced after autoclaving, which does not meet the Chinese Pharmacopoeia and import registration standards. Activated carbon, as a commonly used adsorbent in injections, plays a corresponding role in adsorbing pyrogens. However, the diversity of activated carbon raw materials and production processes, the uncertainty of the activation mechanism, the limitations of quality control, and the possibility of introducing impurities and insoluble particles will bring risks to the use of activated carbon in injections. Due to the diverse sources of activated carbon raw materials and production processes, it may contain different elemental impurities; some elemental impurities are toxic, including neurotoxicity and nephrotoxicity. There are many ways to control pyrogens in injections, and imported injection varieties basically no longer use activated carbon. Therefore, it is an inevitable trend to minimize the risks introduced by activated carbon in injection production.

Patent CN114796110A discloses an ethanol-free insoluble drug concentrate and a micellar solution prepared therefrom. For the insoluble drug nimodipine, egg yolk lecithin, 15-hydroxystearic acid polyethylene glycol ester, and propylene glycol are used in the prescription, wherein the weight percentage of 15-hydroxystearic acid polyethylene glycol ester is as high as 46%, and the weight percentage of propylene glycol is as high as 51%. 15-hydroxystearic acid polyethylene glycol ester is a synthetic excipient with a large dosage and low safety; high concentration of propylene glycol injection can cause pain and irritation, and higher concentrations can cause hemolysis in the human body. The prescription abandons ethanol but still uses organic solvents, and organic solvents themselves have toxic side effects, and their use in injections is more stringent.

Therefore, whether it is possible to provide a safer injection with a higher nimodipine concentration has become a technical problem that needs to be solved urgently in this field.

Summary of the invention

In view of the shortcomings of the above-mentioned technology, the inventors of the present invention provide a high-concentration nimodipine micellar injection for intravenous drip and a preparation method thereof, which is composed of the active ingredient nimodipine and the auxiliary materials phospholipids, bile acid or its salt, surface modification materials, isotonicity regulators, pH regulators and water for injection. The injection not only greatly improves the solubility of nimodipine, but also selects physiologically compatible auxiliary materials phospholipids and bile acid, and a micellar system with good safety for solubilization, adopts an innovative film dispersion-hydration process, and abandons ethanol as a solubilizing agent in the prescription, thereby significantly improving the safety of medication. The injection can be directly diluted with the injection solution, and the compatible solution has good 24h stability, does not require special equipment for infusion pumps and secondary care, and improves medication safety and compliance.

The main inventive concept of the present invention is to provide a more ideal carrier for nimodipine. Based on the physiological compatibility and solubility of nimodipine, the phosphatidylcholine (PC)-bile salt (BS) mixed micelle (MM) preparation is an attractive candidate for intravenous administration of poorly soluble drugs. This simple method of combining water-soluble surfactants with water-insoluble phospholipids can obtain an isotropic and clear solution. The bile salts present in high-concentration bile can dissolve phosphatidylcholine to a large extent, forming a transparent mixed micelle solution, which can then dissolve poorly water-soluble substances. The hemolytic effect of phosphatidylcholine can be neutralized by dissolved phospholipids. Most importantly, the phosphatidylcholine (PC)-bile salt (BS) mixed micelle (MM) preparation has been shown to be well tolerated locally and systemically, with no embryotoxicity after administration, and no teratogenic or mutagenic effects. In the absence of a toxicological solvent mixture, the phosphatidylcholine (PC)-bile salt (BS) mixed micelle (MM) preparation can reduce irritation and improve patient compliance. Therefore, phosphatidylcholine (PC)-bile salt (BS) mixed micelle (MM) preparation is an ideal drug carrier for intravenous administration of nimodipine. Based on the above ideas, this application is ultimately based on the phospholipid-bile salt mixed micelle system, abandons the organic solvent solubilization scheme, develops an innovative formulation design of phospholipids, bile salts and surface modification materials, and constructs a new type of nano-micelle solubilization system to achieve effective solubilization of nimodipine, breaking through the "bottleneck" technical difficulties in the improvement of nimodipine injection.

Specifically, the technical solution of this application is as follows:

A nimodipine micellar injection comprises an active ingredient nimodipine, phospholipids, bile acid or its salt, a surface modification material, an isotonicity regulator, a pH regulator and water for injection; the proportions of the components are as follows:

The concentration of the active ingredient nimodipine in the injection is 0.1 mg/ml to 4 mg/ml, and more preferably 0.5 mg/ml to 2.5 mg/ml; in addition, the weight ratio of nimodipine to phospholipid is 0.5 to 20:100, the weight ratio of bile acid or its salt to phospholipid is 0.1 to 10:1, the weight ratio of the surface modification material to nimodipine is 0 to 10:1, and the weight ratio of the isotonicity regulator to the phospholipid is 0.1 to 5:1; the amount of the pH regulator is determined according to the pH of the final injection;

The volume specification of the above-mentioned nimodipine micellar injection is 1 ml to 10 ml, preferably 5 ml, and is contained in an ampoule bottle.

Preferably, the phospholipids include but are not limited to soybean lecithin, egg yolk lecithin, hydrogenated soybean lecithin (HSPC), hydrogenated egg yolk lecithin (HEPC), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylcholine (DSPC), dilauroylphosphatidylcholine (DLPC), dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylethanolamine (DSPE), and dioleoylphosphatidylcholine (DOPC); soybean lecithin is preferred.

The preferred bile acid is glycocholic acid, deoxycholic acid, glycochenodeoxycholic acid, taurocholic acid and taurochenodeoxycholic acid or a mixture of their salts, or a mixture of the above. Among them, glycocholic acid or sodium glycocholate is preferred.

HS 15或

HS 15)、聚氧乙烯聚氧丙烯共聚物(泊洛沙姆188)、聚氧乙烯脱水山梨醇脂肪酸酯类(聚山梨酯80)。The preferred surface modification material is any one or more of 15-hydroxystearate polyethylene glycol, polyoxyethylene polyoxypropylene copolymer, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene castor oil derivatives, polyethylene glycol vitamin E succinate, polyethylene glycol-distearoylphosphatidylcholine in any proportion; wherein 15-hydroxystearate polyethylene glycol (

HS 15 or

HS 15), polyoxyethylene polyoxypropylene copolymer (Poloxamer 188), polyoxyethylene sorbitan fatty acid esters (Polysorbate 80).

The preferred isotonicity regulator is any one or more of sucrose, trehalose, glucose, lactose, fructose, mannitol, dextran, sorbitol, dextran, glycine, hydroxyethyl starch, polyvinyl pyrrolidone, polyvinyl ketone, etc., in any proportion; sucrose is preferred.

Preferably, the acid in the physiologically and pharmacologically acceptable pH regulator is selected from any one or more of acetic acid, formic acid, trifluoroacetic acid, phosphoric acid, citric acid, tartaric acid, oxalic acid, malic acid, alanine, hydrochloric acid, leucine, isoleucine, malic acid, valine, tryptophan, maleic acid, fumaric acid, lactic acid, phenylalanine, methionine and succinic acid in any proportion, wherein citric acid is preferred; the base in the pH regulator is any one or more of sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium hydroxide, choline hydroxide, arginine, lysine, histidine, diethylamine, triethylamine, meglumine, sodium citrate, sodium tartrate, sodium lactate, sodium phosphate, and disodium hydrogen phosphate in any proportion, wherein sodium hydroxide is preferred.

In addition, the inventors also provide a method for preparing the above-mentioned nimodipine micelle injection, which is a thin film dispersion method, and the specific process is as follows:

Step (1): weighing nimodipine, phospholipid, bile acid or its salt, and surface modification material in the formulation, and dissolving them in an appropriate amount of organic solvent;

Step (2): transfer the solution obtained in step (1) to a rotary evaporator, set the water bath temperature at 30-70°C, open the vacuum to remove the organic solvent, and obtain a loose film;

Step (3): weigh the isotonicity regulator in the formulation and dilute it in water for injection; if the excipient in step (1) is bile acid, add the base in the pH regulator at a molar ratio of 1:1 to the bile acid, fill with protective gas, and obtain a stabilizer solution; transfer the stabilizer solution completely to the loose film obtained in step (2), stir to fully hydrate it, adjust the pH of the solution with a pH regulator, heat treat, and cool to obtain a nimodipine micellar solution;

Step (4): the solution obtained in step (3) is made up to volume with water for injection, filled with protective gas, sterilized and filtered, divided into ampoules, sealed, and sterilized by hot pressing to obtain nimodipine micellar injection.

In certain preferred embodiments, the present invention relates in part to a nimodipine micellar injection, in which the ratio of bile acid or its salt to phospholipid is 0.5-2:1; the weight ratio of the surface modification material to nimodipine is 0-10:1, and as one of the preferred embodiments, it is preferably 0.1:1.

The above-mentioned thin film dispersion method organic solvent is any one or more of methanol, ethanol, ethyl acetate, and isopropanol in any proportion. The amount of organic solvent is 2% to 8% of the volume of the prepared injection solution. The protective gas is any one of nitrogen, helium, carbon dioxide and argon.

The preferred organic solvent is a mixed solvent of methanol and ethanol, and the preferred ratio of methanol to ethanol is 4:1.

The heat treatment described in step (3) is at 90-100° C. and the time is 20-30 min. The protective gas described in steps (3) and (4) is nitrogen, and the time for passing the protective gas is 0.5-2 hours, wherein step (4) needs to control the dissolved oxygen residual range to 5-10 ppm.

The specification of the nanomicelle injection obtained in step (4) is 0.1 mg/ml to 4 mg/ml, and more preferably 0.5 mg/ml to 2.5 mg/ml.

The pH value of the micelle injection finally obtained in the above step (4) is 5.0-8.5, preferably 5.5-7.5; the Z-Average of the micelle injection is 2-10nm; the Zeta potential of the micelle injection is -20mV--50mV; the transmittance of the micelle injection is greater than 90%.

In specific applications, the dilution injections that can be used in combination with the nimodipine micellar injection provided by the present invention include, but are not limited to, 0.9% sodium chloride injection, 5% glucose, sodium lactate Ringer's solution, magnesium-containing sodium lactate Ringer's solution, dextran 40 solution, 6% polyoxy-2-hydroxyethyl starch, 5% human albumin or blood, and as one of the preferred embodiments, 0.9% sodium chloride injection or 5% glucose is preferred. When diluted to a concentration of 0.02 mg/ml to 0.04 mg/ml, it is still a clear and transparent solution and no precipitated crystals of nimodipine are observed.

The nimodipine micellar injection provided by the present invention can be used to treat cerebral vasospasm after subarachnoid hemorrhage due to various reasons, such as but not limited to, improving blood circulation in the recovery period of acute cerebrovascular disease, treating diffuse brain injury together with hyperbaric oxygen therapy, assisting recovery after cranial nerve injury, treating frequent migraine and preventing migraine, peripheral vertigo and Meniere's disease, reducing the occurrence of postoperative delirium in elderly patients under general anesthesia, drug-resistant epilepsy, orgasm headache and bathing-related headache.

Compared with the prior art, the present invention has the following beneficial effects:

(1) The nimodipine micellar injection prepared by the present invention can selectively add an auxiliary material containing a polyethylene glycol chain as a micelle surface modification material, specifically 15-hydroxystearate polyethylene glycol ester (Solutol HS 15 or Kolliphor HS 15), polyoxyethylene polyoxypropylene copolymer (Poloxamer 188), polyoxyethylene sorbitan fatty acid esters (polysorbate 80) or polyethylene glycol 2000-distearate phosphatidylethanolamine (PEG2000-DSPE), polyoxyethylene castor oil (Cremophor ELP or Kolliphor ELP), polyethylene glycol vitamin E succinate (TPGS 1000); when selected for addition, the dosage is small, and the weight percentage can be as low as 0.02% of the whole system. A new type of nano-micelle solubilization system can be constructed, which can significantly improve the physical and chemical stability of the mixed micelles, increase the drug loading, and greatly improve the water solubility and stability of nimodipine. After low-temperature cycle test, freeze-thaw test and compatibility stability test, no crystal precipitation was found. The stability of electrolyte solutions such as 0.9% sodium chloride injection is significantly improved, so that electrolyte injection can be used for infusion, which is convenient for clinical application.

(2) The nimodipine micellar injection prepared by the present invention has an injection concentration of up to 4 mg/ml, which solves the bottleneck problem of poor solubility of nimodipine injection and has higher clinical value than the commercially available injection of 0.2 mg/ml.

(3) The nimodipine micellar injection prepared by the present invention adds a heat treatment step during the preparation process, which significantly improves the physical and chemical stability of the injection. After filtration, the drug solution is placed at 25° C. for 48 hours, and there is no significant change in appearance, content, related substances, particle size, etc., which provides sufficient time for the next steps of filling and sealing, which is conducive to scale-up production.

(4) The nimodipine micellar injection prepared by the present invention can withstand hot pressure sterilization. There is no significant change in the properties, pH, particle size, content, related substances and other indicators of the injection before and after sterilization, which meets the requirements of the 2020 edition of the Pharmacopoeia and greatly improves the sterility assurance.

(5) The nimodipine micellar injection prepared by the present invention does not contain ethanol solubilizers and organic solvents, which reduces allergic reactions, alcohol poisoning or impaired alcohol metabolism, pain and inflammation, and significantly improves the safety of clinical medication.

(6) The preparation process described in the present invention is easy to scale up, and adopts an innovative thin film dispersion-hydration process, which significantly reduces the amount of organic solvents such as anhydrous ethanol, and abandons the use of activated carbon in the process, which is conducive to scale-up production. However, the micelle patents in the prior art use anhydrous ethanol or propylene glycol, which have low safety and large usage, increasing the difficulty of scale-up production; other complex injection forms such as fat emulsions and liposomes have complex prescription processes, which are not conducive to process scale-up, are difficult to control quality, and have poor stability.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the appearance of the nimodipine mixed micelle injection prepared by the thin film dispersion method in Example 1.

FIG. 2 shows a high performance liquid chromatogram of the determination of related substances in the nimodipine micellar injection prepared by the thin film dispersion method in Example 5.

FIG3 and FIG4 show the particle size distribution diagrams of the nimodipine micellar injection prepared by the thin film dispersion method in Example 7.

FIG5 shows the content change curve (0.02 mg/ml and 0.04 mg/ml) of the nimodipine micellar injection prepared by the thin film dispersion method in Example 7 of Experimental Example 8 during the compatibility stability study (0.9% sodium chloride injection).

FIG6 shows the content change curve (0.02 mg/ml and 0.04 mg/ml) of the nimodipine micellar injection prepared by the thin film dispersion method in Example 7 of Experimental Example 8 during the compatibility stability study (5% glucose injection).

FIG. 7 is a curve of the content change (0.02 mg/ml and 0.04 mg/ml) measured during the compatibility stability study (0.9% sodium chloride injection) of the original injection (Nimotop) in Experimental Example 8.

FIG. 8 is a curve of content changes (0.02 mg/ml and 0.04 mg/ml) measured during the compatibility stability study (5% glucose injection) of the original injection (Nimotop) in Experimental Example 8.

DETAILED DESCRIPTION

The above contents of the present invention are further described in detail below through specific implementation methods in the form of examples, but this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies implemented based on the above contents of the present invention belong to the scope of the present invention, and unless otherwise specified, the following examples are all completed using conventional existing technologies.

Example 1 Preparation of Nimodipine Micellar Injection

Each vial contains 10 mg of nimodipine. The composition of 25 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, soybean lecithin, and glycocholic acid according to the prescribed amount, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation to form a thin film;

(3) Sucrose, sodium hydroxide, citric acid, and sodium citrate were weighed according to the above prescription, dissolved in 100 ml of water for injection to obtain a clear solution, filled with nitrogen as a protective gas, and added to the above film to completely hydrate to obtain a clear liquid, heat treated in an oil bath at 100° C. for 30 min, cooled to room temperature, adjusted the pH value to 6.5 with 0.1 mol/L sodium hydroxide solution, and diluted to 125 ml with water for injection to obtain a nimodipine mixed micellar solution;

(4) Sterile filtration, packaging into ampoules, sealing, and autoclaving at 121°C for 15 min to obtain nimodipine mixed micelle injection.

As shown in Figure 1, the injection solution of the same batch was clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution was 2.577 nm, the PDI was 0.239, and the Zeta potential was -30.0 mV.

Example 2

Each vial contains 10 mg of nimodipine. The composition of 25 vials of nimodipine micellar injection is as follows:

Preparation process:

Steps (1) and (2) are the same as in Example 1.

(3) Weigh sucrose and sodium hydroxide according to the above prescription, dissolve them in 100 ml of water for injection to obtain a clear solution, fill with nitrogen as a protective gas, add to the above film, hydrate completely to obtain a clear liquid, adjust the pH value to 6.5 with 0.1 mol/L sodium hydroxide solution, and dilute to 125 ml with water for injection to obtain a nimodipine mixed micellar solution;

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121°C for 12 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 5.105 nm, the PDI is 0.350, and the Zeta potential is -43.9 mV.

Example 3

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

Steps (1) and (2) are the same as in Example 1.

(3) According to the above prescription, weigh sucrose and dissolve it in 80 ml of water for injection to obtain a clear solution. Fill with nitrogen as a protective gas and add it to the above film. After complete hydration, a clear liquid is obtained. Heat-treat in an oil bath at 80°C for 30 min and cool to room temperature. Adjust the pH value to 6.0 with 0.1 mol/L sodium hydroxide solution and dilute to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 12 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 2.639 nm, the PDI is 0.301, and the Zeta potential is -30.5 mV.

Example 4

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

Steps (1) and (2) are the same as in Example 1.

(3) According to the above prescription, sucrose and sodium hydroxide were weighed and dissolved in 80 ml of water for injection to obtain a clear solution, which was filled with nitrogen as a protective gas and added to the above film. After complete hydration, the film was heat-treated in an oil bath at 100°C for 60 min and cooled to room temperature. The pH value was adjusted to 6.0 with 0.1 mol/L citric acid solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 12 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 5.698nm, the PDI is 0.310, and the Zeta potential is -29.7mV.

Example 5

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, soybean lecithin, glycocholic acid, and poloxamer 188 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation to form a thin film;

(3) According to the above prescription, sucrose, sodium hydroxide and citric acid were weighed and dissolved in 80 ml of water for injection, and added to the above film. After complete hydration, a clear solution was obtained. The solution was heat-treated in an oil bath at 100°C for 30 min and cooled to room temperature. The pH value was adjusted to 6.5 with 0.1 mol/L citric acid solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 12 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. As shown in Figures 3 and 4, the Z-Average of the mixed micelle solution is 2.734 nm, the PDI is 0.255, and the Zeta potential is -28.8 mV.

Example 6

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, soybean lecithin, glycocholic acid, and Solutol HS 15 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation to form a thin film;

(3) According to the above prescription, sucrose, sodium hydroxide, citric acid and sodium citrate were weighed and dissolved in 80 ml of water for injection to obtain a clear solution. The solution was filled with nitrogen for 10 min and added to the above film. After complete hydration, the pH value was adjusted to 6.5 with 0.1 mol/L sodium hydroxide solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micellar solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 15 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 9.860nm, the PDI is 0.357, and the Zeta potential is -26.9mV.

Example 7

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, soybean lecithin, glycocholic acid, and Solutol HS 15 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation to form a thin film;

(3) According to the above prescription, sucrose, sodium hydroxide and citric acid were weighed and dissolved in 80 ml of water for injection to obtain a clear solution. The solution was filled with nitrogen for 30 min and added to the above film. After complete hydration, the solution was heat treated in an oil bath at 100°C for 30 min and cooled to room temperature. The pH value was adjusted to 6.5 with 0.1 mol/L sodium hydroxide solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 15 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 2.834 nm, the PDI is 0.201, and the Zeta potential is -34.7 mV.

Example 8

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, soybean lecithin, glycocholic acid, and Solutol HS 15 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation to form a thin film;

(3) According to the above prescription, sucrose, sodium hydroxide and citric acid were weighed and dissolved in 80 ml of water for injection to obtain a clear solution. The solution was filled with nitrogen for 60 min and added to the above film. After complete hydration, the solution was heat treated in an oil bath at 100°C for 30 min and cooled to room temperature. The pH value was adjusted to 6.5 with 0.1 mol/L citric acid solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 15 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 3.325 nm, the PDI is 0.315, and the Zeta potential is -37.6 mV.

Example 9

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, egg yolk lecithin, taurocholic acid, and Solutol HS 15 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation until all methanol and ethanol are removed and a thin film is formed;

(3) According to the above prescription, sucrose and sodium hydroxide were weighed and dissolved in 80 ml of water for injection to obtain a clear solution. The solution was filled with nitrogen for 60 min and added to the above film. After complete hydration, the film was heat treated in an oil bath at 100°C for 30 min and cooled to room temperature. The pH value was adjusted to 5.5 with 0.1 mol/L citric acid solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 15 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 3.325 nm, the PDI is 0.315, and the Zeta potential is -37.6 mV.

Example 10

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, hydrogenated soybean lecithin, glycocholic acid, and poloxamer 188 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) placing the clear solution obtained in step (1) on a rotary evaporator to remove methanol and ethanol until they are completely removed and a thin film is formed;

(3) According to the above prescription, sucrose and sodium hydroxide were weighed and dissolved in 60 ml of water for injection to obtain a clear solution. The solution was filled with nitrogen for 30 min and added to the above film. After complete hydration, the film was heat treated in an oil bath at 100°C for 30 min and cooled to room temperature. The pH value was adjusted to 6.5 with 0.1 mol/L sodium hydroxide solution and the volume was adjusted to 80 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 15 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 2.679nm, the PDI is 0.371, and the Zeta potential is -29.1mV.

Embodiment 11

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, soybean lecithin, glycocholic acid, and Solutol HS 15 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) placing the clear solution obtained in step (1) on a rotary evaporator to remove methanol and ethanol until they are completely removed and a thin film is formed;

(3) According to the above prescription, sucrose and sodium hydroxide were weighed and dissolved in 80 ml of water for injection to obtain a clear solution. The solution was filled with nitrogen for 30 min and added to the above film. After complete hydration, the film was heat treated in an oil bath at 100°C for 30 min and cooled to room temperature. The pH value was adjusted to 7.0 with 0.1 mol/L sodium hydroxide solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 15 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 3.517nm, the PDI is 0.392, and the Zeta potential is -34.2mV.

Example 12

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) Weigh nimodipine, soybean lecithin, glycocholic acid, and polysorbate 80 according to the ratio, dissolve them in methanol and ethanol, and stir to disperse them evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation until all methanol and ethanol are removed and a thin film is formed;

(3) According to the above prescription, mannitol and sodium hydroxide were weighed and dissolved in 80 ml of water for injection to obtain a clear solution, which was filled with nitrogen as a protective gas and added to the above film. After complete hydration, the film was heat-treated in an oil bath at 100°C for 30 min and cooled to room temperature. The pH value was adjusted to 6.5 with 0.1 mol/L citric acid solution and the volume was adjusted to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 12 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 2.267nm, the PDI is 0.321, and the Zeta potential is -31.5mV.

Example 13

Each vial contains 10 mg of nimodipine. The composition of 20 vials of nimodipine micellar injection is as follows:

Preparation process:

(1) According to the above prescription, nimodipine, soybean lecithin, glycocholic acid, and polyethylene glycol 1000 vitamin E succinate (TPGS1000) were weighed and dissolved in methanol and ethanol, and stirred to disperse evenly to obtain a clear liquid;

(2) removing methanol and ethanol from the clear solution obtained in step (1) by rotary evaporation until all methanol and ethanol are removed and a thin film is formed;

(3) According to the above prescription, weigh trehalose and sodium hydroxide and dissolve them in 80 ml of water for injection to obtain a clear solution. Fill with nitrogen as protective gas and add it to the above film. After complete hydration, heat treat it in an oil bath at 100°C for 30 min and cool it to room temperature. Adjust the pH value to 6.0 with 0.1 mol/L citric acid solution and make the volume to 100 ml with water for injection to obtain a nimodipine mixed micelle solution.

(4) sterilizing and filtering, packaging into ampoules, sealing, and sterilizing by hot pressing at 121° C. for 12 min to obtain nimodipine mixed micelle injection.

The injection solution is clear, transparent and light yellow-green. The Z-Average of the mixed micelle solution is 2.627nm, the PDI is 0.291, and the Zeta potential is -35.1mV.

The above embodiments have been verified by experiments, and Embodiment 5-13 can be preferably applied, and Embodiment 5-8 is most preferred.

Comparative Example 1

According to Example 1 in patent CN 102525917B, nimodipine micellar injection was prepared in the following steps:

Weigh 50 mg of egg yolk lecithin, 50 mg of sodium glycocholate, and 1.25 mg of nimodipine into a 50 ml round-bottom flask, add 10 ml of ethanol, dissolve, and disperse evenly by ultrasonication. Remove ethanol by rotary evaporation at a water bath temperature of 40°C until there is no alcohol smell and a transparent film is formed, and then disperse with 2.5 ml of injection water to obtain a dispersion solution of drug-containing mixed micelles. Add 0.05% injection-grade activated carbon and stir for 15 minutes, centrifuge at 12000 rpm/min for 5 minutes, filter with a 0.22 μm microporous filter membrane, take the filtrate, divide it into portions, and sterilize it by hot pressing at 121°C for 15 minutes to obtain nimodipine mixed micelle injection.

Comparative Example 2

According to Example 2 in patent CN 102525917B, nimodipine micellar injection was prepared in the following steps:

Weigh 50 mg of egg yolk lecithin, 50 mg of sodium glycocholate, and 1 mg of nimodipine into a 50 ml round-bottom flask, add 10 ml of ethanol, dissolve, and disperse evenly by ultrasonication. Then remove the ethanol by rotary evaporation, and the water bath temperature is 35°C. Rotate until there is no alcohol smell and a transparent film is formed. Then disperse with 2.5 ml of injection water to obtain a dispersion solution of drug-containing mixed micelles. Add 0.05% injection-grade activated carbon and stir for 15 minutes, centrifuge at 12000 rpm/min for 5 minutes, and then filter with a 0.22 μm microporous filter membrane, take the filtrate, divide it into packages, and sterilize it by hot pressing at 121°C for 15 minutes to obtain nimodipine mixed micelle injection.

Experimental Example 1 Determination of drug loading, content, related substances and residual solvents

1. Drug loading = (amount of nimodipine dissolved in the prescription)/{amount of total excipients added (phospholipids + bile salts + nimodipine)}.

2. Content determination method

Chromatographic conditions: octadecylsilane bonded silica gel as filler; methanol-acetonitrile-water (35:38:27) as mobile phase; detection wavelength at 235 nm; injection volume 10 μl, column temperature 40°C.

Reference solution: Weigh 20 mg of nimodipine, place in a 20 ml volumetric flask, add methanol to dissolve and dilute to the scale, shake well, dilute 10 times with solvent (methanol-acetonitrile-water, 35:38:27), shake well to obtain.

Test solution: Accurately measure 2 ml of the test solution under the relevant substance, place it in a 20 ml volumetric flask, dilute to the scale with solvent (methanol-acetonitrile-water, 35:38:27), and shake well.

3. Test methods for related substances

Chromatographic conditions: octadecylsilane bonded silica gel as filler; methanol-water (40:60) as mobile phase A, acetonitrile-water (90:10) as mobile phase B; detection wavelength 235nm; injection volume 10μl, column temperature 40°C. Gradient elution according to Table 1.

Table 1 Related substances of nimodipine micellar injection Method gradient elution table

Test solution: Accurately measure 1 ml of the product, place it in a 10 ml volumetric flask, dilute to the scale with methanol, and shake well.

4. Residual solvent determination method:

(1) Chromatographic conditions: capillary column with 6% cyanopropylphenyl-94% dimethylpolysiloxane (or similar polarity) as stationary phase (Agilent DB-624, 0.53 mm × 30 m, 3 μm or equivalent chromatographic column is recommended); initial temperature is 40°C, maintained for 8 min, raised to 180°C at a rate of 10°C per minute, maintained for 5 min; injection port temperature is 200°C; detector is flame ionization detector, detector temperature is 300°C; column flow rate is 2 ml per minute; split ratio is 10:1; headspace equilibrium temperature is 80°C; quantitative loop temperature is 90°C; equilibrium time is 30 min.

System suitability requirements: In the chromatogram of the reference solution, methanol and ethanol appear in sequence, and the separation between each chromatographic peak should meet the requirements.

(2) Solution preparation

Test solution: Take about 100 mg of the product, weigh accurately, place in a 20 ml headspace bottle, accurately add 2 ml of ultrapure water, shake, and seal.

Reference solution: Take appropriate amounts of methanol and ethanol respectively, weigh them accurately, and quantitatively dilute them with ultrapure water to make a mixed solution containing approximately 150μg and 250μg of each per 1ml. Accurately measure 2ml of the solution, place it in a 20ml headspace bottle, and seal it.

According to the above-mentioned drug loading, content, related substances, and residual solvent determination methods, Examples 1-13 and Comparative Examples 1-2 were tested, and the results are shown in Table 2; the chromatogram of related substances of the micelle injection prepared in Example 5 after sterilization is shown in Figure 2. It can be seen from the results in Table 2 that the drug loading of the micelle injection prepared in the example is significantly higher than that of the comparative example; the content of the micelle injection prepared in the example has no significant change before and after hot pressure sterilization, and the related substances meet the pharmacopoeia and import registration standards; the content of the injection prepared in Comparative Examples 1-2 decreased significantly before and after sterilization, and the related substances exceeded the standards, so Example 1-13 has a significant advantage.

Table 2 Test results of drug loading, content, related substances, etc. of nimodipine micellar injection

Experimental Example 2: Investigating the Effect of Adding Surface Modification Materials

The nimodipine micelle injection prepared by the invention has a surface modification material added into the prescription to construct a novel nano-micelle solubilization system, which can significantly improve the physical and chemical stability of the mixed micelles, increase the drug loading, and greatly improve the water solubility and stability of nimodipine.

From the drug loading results of Experimental Example 1, it can be seen that no surface modification material was added to the prescriptions of Examples 1-4, and their drug loadings were all greater than those of the comparative example. Different prescription amounts of surface modification materials were added to Examples 5-11, and their drug loadings were all greater than those of Examples 1-4. Adding surface modification materials can increase the drug loading.

By comparing Example 2 (without addition) and Example 6 (with addition), neither of which was heat treated, the sample was placed at 25°C for 48 hours after filtration, and the physical stability was judged by observing the properties (whether crystals were precipitated). The results are shown in the following table. Example 2 precipitated after being placed for 6 hours, and the injection prepared in Example 6 was placed at 25°C for 12 hours without precipitation, which can significantly improve the physical stability of the micelle injection.

Table 3 Physical stability test results

Experimental Example 3: Effect of heat treatment on stability

The nimodipine micellar injection prepared by the present invention has a heat treatment step added during the preparation process, and embodiments 2, 3, 4, 5, and 7 are designed for investigation and compared with the comparative example. The filtered samples are placed at 25° C. for 48 hours, and the physical stability is judged by observing the properties (whether crystals are precipitated).

From the results in Table 4, it can be seen that the samples of Comparative Examples 1-2 were filtered and placed at 25°C for 4 hours, and crystals were precipitated; Example 2 was placed for 6 hours and crystals were precipitated, which was better than the comparative example; Example 3 was placed for 8 hours and crystals were precipitated; Examples 4, 5 and 7 were placed at 25°C for 36 hours and the properties did not change, and Examples 5 and 7 were placed for 48 hours and still did not precipitate;

The content and related substances were tested according to the method in Experimental Example 1. From the results in Table 5 below, it can be seen that there was no significant change in pH, content, related substances, etc.

Based on the above experimental results, the nimodipine micellar injection prepared by the present invention significantly improves the physical stability of the injection by adding a heat treatment step during the preparation process. The filtered solution is placed at 25°C for 48 hours, and there is no significant change in appearance, content, related substances, particle size, etc.

Table 4 Heat treatment results

Table 5 Test results of content, related substances, particle size, etc. of nimodipine micellar injection

Experimental Example 4: Investigating the Effect of Protective Gas on Micelle Injection

Comparative Examples 5-8, the necessity of filling with protective gas can be seen from the results in Table 6, and the filling time of protective gas is maintained for 0.5-1h. The use of nitrogen as protective gas in the hydration solution of nimodipine mixed micelle injection can reduce the level of related substances (key impurity I) and improve the physical and chemical stability of nimodipine injection.

Table 6 Test results of related substances

Experimental Example 5 Investigation of Sterilization Conditions

The influence of hot-pressing sterilization conditions on the content of nimodipine micelle injection was investigated, and the prepared micelle injection was sterilized by hot-pressing at 121°C/12min or 121°C/15min, respectively, and the content detection method in Example 6 was used, and the results are shown in Table 7. It can be seen from Table 7 that there is no significant change in the content results before and after sterilization. Considering the stability of the excipients and other factors, hot-pressing sterilization at 121°C/12min was selected.

Table 7 Test results of homemade samples under different sterilization conditions

Experimental Example 6 Low Temperature Cycle Test

The nimodipine micellar injection prepared in Example 5 was subjected to a low-temperature cycle test. The test should include 3 cycles, each cycle was placed at 2-8°C for 2 days, and then placed at 40°C for 2 days. After each cycle, samples were taken for testing, and the content and related substances were tested according to the methods in Experimental Example 1. The results are shown in Table 8.

From the results in Table 8, it can be seen that after the three rounds, there was no precipitation phenomenon; the pH value, clarity and color, particle size, potential, content, related substances, etc. had no significant changes compared with day 0, and the homemade micelle injection solution could withstand the low-temperature cycle test.

Table 8 Low temperature cycle test results

Experimental Example 7 Freeze-thaw test

The nimodipine micellar injection prepared in Example 4 was subjected to a freeze-thaw test. The test should include 3 cycles, each cycle was placed at -10--20°C for 2 days, and then placed at 40°C for 2 days. After each cycle, samples were taken for testing. The content and related substances were tested according to the method in Experimental Example 1. The results are shown in Table 9.

From the results in Table 9, it can be seen that after 3 cycles, no precipitation was observed; the pH value, clarity and color, particle size, potential, content, related substances, etc. had no significant changes compared with day 0, and the homemade micelle injection could withstand the freeze-thaw test.

Table 9 Freeze-thaw test results

Experimental Example 8 Compatibility Stability

分别用0.9％氯化钠注射液、5％葡萄糖注射液稀释至浓度为0.04mg/ml或0.02mg/ml，室温(25℃)下放置24h，考察临床使用条件下的稳定性。结合实验例1中含量、有关物质分析检测方法，对稀释稳定性进行考察，结果见表10，表11，含量变化曲线见附图5-8。Take the nimodipine micellar injection prepared in Example 7 and the original injection

Diluted to a concentration of 0.04 mg/ml or 0.02 mg/ml with 0.9% sodium chloride injection and 5% glucose injection respectively, placed at room temperature (25°C) for 24 hours, and investigated the stability under clinical use conditions. Combined with the content and related substance analysis and detection methods in Experimental Example 1, the dilution stability was investigated, and the results are shown in Table 10 and Table 11, and the content change curves are shown in Figures 5-8.

From the results in Tables 10 and 11, it can be seen that no precipitation was observed in the self-made product (Example 7) within 24 hours, the content and related substances (impurity I and total impurities) did not change significantly, and the insoluble particles did not increase significantly.

Table 10 Example 7 Compatible solution content determination results

Table 11 Results of content determination of original injection compatibility solution

稀释于0.9％氯化钠注射液或5％葡萄糖注射液，含量波动显著，尤其是稀释于0.9％氯化钠注射液放置1h后波动异常，分析原因为配伍溶液有结晶析出，进样采集到析出晶体导致含量测试异常，无需考察后续稳定性。相对于原研品注射液，自制样品的配伍溶液放置24h无析出现象，且含量无显著变化，稳定性好。Conclusion: Original injection

The content fluctuated significantly after dilution in 0.9% sodium chloride injection or 5% glucose injection, especially after being diluted in 0.9% sodium chloride injection for 1 hour. The reason for the abnormal fluctuation was that the compatible solution had crystals precipitated, and the precipitated crystals were sampled and collected, resulting in abnormal content test. There was no need to investigate the subsequent stability. Compared with the original product injection, the compatible solution of the homemade sample had no precipitation after being placed for 24 hours, and the content did not change significantly, with good stability.

Experimental Example 9 Influencing Factors Test

The nimodipine micellar injection prepared in Example 8 was placed at a high temperature of 40°C and under illumination conditions (illumination intensity of 4500 lx, ultraviolet 200 W·h/m ² ) to investigate the influencing factors, and samples were taken on the 30th day, and the content and related substances were detected according to the method of Experimental Example 1. The results are shown in Table 12. From the results in Table 12, it can be seen that the homemade injection was placed at a high temperature of 40°C and under illumination conditions for 30 days, and its properties, pH value, clarity and color, particle size, potential, content, and related substances did not change significantly, and all met the pharmacopoeia standards.

Table 12 Test results of influencing factors

Experimental Example 10 Accelerated test and long-term test

The stability of the nimodipine mixed micelle injection in Examples 11, 12 and 13 was studied under the following conditions, and the changes in appearance, properties, pH and Zeta potential were recorded. The nimodipine content and related substances were determined according to Example 6 above. The results are shown in Table 13.

Among them, the content of the control sample decreased by about 6% after sterilization, and the total impurities exceeded the standard in the pharmacopoeia, so the stability test was not carried out.

Long-term test: 25℃±2℃/60%RH±5%RH; Accelerated test: 40℃±2℃/75%RH±5%RH.

Table 13 Results of stability study of nimodipine micellar injection

Conclusion: It can be seen from the above table that the nimodipine micellar injection of Examples 11-13 was placed under the conditions of long-term stability and accelerated stability for 6 months, and the appearance, pH, potential, content, related substances (impurity I, total impurities) and other indicators did not change significantly. The above results show that the nimodipine micellar injection of Examples 11-13 has good stability. The storage conditions of the nimodipine micellar injection are designed to be stored at 2-8°C. According to the results of the accelerated stability experiment, the shelf life of the nimodipine micellar injection is tentatively set at 2 years.

For ordinary technicians in this field, the specific embodiments are only illustrative descriptions of the present invention. It is obvious that the specific implementation of the present invention is not limited to the above-mentioned methods. As long as various non-substantial improvements are made using the method concepts and technical solutions of the present invention, or the concepts and technical solutions of the present invention are directly applied to other occasions without improvement, they are all within the protection scope of the present invention.

Claims (10)

1. A nimodipine micelle injection is characterized in that: comprises nimodipine, phospholipid, cholic acid or its salt, surface modification material, isotonic regulator, pH regulator and water for injection.

2. Nimodipine micelle injection according to claim 1, wherein: the proportions of the components are as follows:

the concentration of nimodipine as the active component in the injection is 0.1 mg/ml-4 mg/ml; in addition, the weight ratio of nimodipine to phospholipid is 0.5-20: 100, the weight ratio of cholic acid or salt thereof to phospholipid is 0.1-10: 1, the weight ratio of the surface modification material to nimodipine is 0-10: 1, the weight ratio of the isotonic regulator to the phospholipid is 0.1-5: 1.

3. nimodipine micelle injection according to claim 1, wherein:

the phospholipid is selected from one of soybean phospholipid, yolk phospholipid, hydrogenated soybean phospholipid, hydrogenated yolk phospholipid, dipalmitoyl phosphatidylcholine, dipalmitoyl phosphatidylethanolamine, distearoyl phosphatidylcholine, dilauroyl phosphatidylcholine, dioleoyl phosphatidylethanolamine, distearoyl phosphatidylethanolamine and dioleoyl phosphatidylcholine.

4. Nimodipine micelle injection according to claim 1, wherein:

the cholic acid or its salt is one or more of glycocholic acid, deoxycholic acid, glycochenodeoxycholic acid, taurocholic acid and taurochenodeoxycholic acid or its salt.

5. Nimodipine micelle injection according to claim 1, wherein:

the surface modification material is composed of any one or more of 15-hydroxystearic acid polyethylene glycol ester, polyoxyethylene polyoxypropylene copolymer, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene castor oil derivatives, polyethylene glycol vitamin E succinate and polyethylene glycol-distearoyl phosphatidylcholine according to any proportion.

6. Nimodipine micelle injection according to claim 1, wherein:

the isotonic regulator is one or more of sucrose, trehalose, glucose, lactose, fructose, mannitol, dextran, sorbitol, dextran, glycine, hydroxyethyl starch, polyvinylpyrrolidone and polyvinylpyrrolidone according to any proportion;

the acid in the physiologically and pharmaceutically acceptable pH regulator is selected from any one or more of glacial acetic acid, formic acid, trifluoroacetic acid, phosphoric acid, citric acid, tartaric acid, oxalic acid, malic acid, alanine, hydrochloric acid, leucine, isoleucine, malic acid, valine, tryptophan, maleic acid, fumaric acid, lactic acid, phenylalanine, methionine and succinic acid according to any proportion; the alkali in the pH regulator is one or more of sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium hydroxide, choline hydroxide, arginine, lysine, histidine, diethylamine, triethylamine, meglumine, sodium citrate, sodium tartrate, sodium lactate, sodium phosphate and disodium hydrogen phosphate according to any proportion.

7. Nimodipine micelle injection according to any one of claims 1-6, wherein:

the phospholipid is selected from soybean phospholipid, and the cholic acid or salt thereof is selected from glycocholic acid or sodium glycocholate; the surface modification material is one of 15-hydroxystearic acid polyethylene glycol ester, polyoxyethylene polyoxypropylene copolymer and polyoxyethylene sorbitan fatty acid ester, the isotonic regulator is sucrose, the acid in the pH regulator is selected from citric acid, and the alkali in the pH regulator is sodium hydroxide.

8. The method for preparing nimodipine micelle injection as claimed in claim 1, which is characterized in that: the preparation method is a film dispersion method, and the specific process is as follows:

step (1): weighing nimodipine, phospholipid, cholic acid or salt thereof and surface modification material in the formula, and dissolving in a proper amount of organic solvent;

step (2): transferring the solution obtained in the step (1) to a rotary evaporator, and opening vacuum to remove the organic solvent at the water bath temperature of 30-70 ℃ to obtain a loose film;

step (3): weighing the isotonic regulator in the formula, and diluting the regulator in water for injection; if the auxiliary material in the step (1) is cholic acid, adding alkali in a pH regulator with the molar ratio of the alkali to the cholic acid being 1:1, and filling a protective gas to obtain a stabilizer solution; completely transferring the nimodipine micelle solution into the loose film obtained in the step (2), stirring to enable the loose film to be hydrated completely, adjusting the pH of the solution by a pH regulator, performing heat treatment, and cooling to obtain nimodipine micelle solution;

Step (4): and (3) fixing the volume of the solution obtained in the step (3) by using water for injection, filling protective gas, sterilizing, filtering, sub-packaging into ampoule bottles for sealing, and performing hot-pressing sterilization to obtain nimodipine micelle injection.

9. The method for preparing nimodipine micelle injection according to claim 8, wherein:

the organic solvent in the film dispersion method is one or a plurality of compositions of methanol, ethanol, ethyl acetate and isopropanol according to any proportion, the dosage of the organic solvent is 2-8% of the volume of the prepared injection, and the protective gas is one of nitrogen, helium, carbon dioxide and argon;

the heat treatment in the step (3) is carried out at the temperature of 90-100 ℃ for 20-30 min; the protective gas in the step (3) and the step (4) is nitrogen, the time of introducing the protective gas is 0.5-2 hours, and the residual range of dissolved oxygen in the step (4) is controlled to be 5-10 ppm;

the concentration of nimodipine in the nano micelle injection obtained in the step (4) is 0.1 mg/ml-4 mg/ml.

10. The use of nimodipine micelle injection according to claim 1 as a medicament for preventing and treating ischemic nerve injury caused by cerebral vasospasm after aneurysmal subarachnoid hemorrhage.

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