Application of solubilizer in levornidazole derivative fat emulsion injection

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of a solubilizer in a levornidazole derivative fat emulsion injection, and the solubilizer is selected from one or more of isopropyl myristate or isopropyl palmitate. The L-ornidazole derivative fat emulsion injection with higher safety and better stability is obtained by applying a solubilizer in the L-ornidazole derivative fat emulsion injection, irritation generated during injection can be reduced, terminal sterilization can be realized, the safety risk of a sample is greatly reduced, the drug loading capacity is also greatly increased, and the injection quality of the L-ornidazole derivative fat emulsion injection is improved. And the clinical use is more convenient.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to application of a solubilizer in a levornidazole derivative fat emulsion injection. Background Art

[0002] Levoornidazole is the levorotatory form of ornidazole, a third-generation nitroimidazole drug developed by Nanjing Shenghe. Its clinical efficacy against anaerobic infections is comparable to that of ornidazole, and its overall incidence of adverse reactions is significantly reduced, only 1 / 15 of that of ornidazole, making it safer for clinical use. Levoornidazole sodium chloride injection developed by Nanjing Shenghe Pharmaceutical Co., Ltd. was approved for marketing in 2009. However, due to the extremely poor water solubility of levoornidazole, acid is usually added to enhance the water solubility of the drug, resulting in an infusion preparation with a pH of about 3.5, which is high in acidity and exceeds the human body's tolerance pH of 4.0 to 9.0. It is highly irritating during injection, causing clinical compliance problems such as instillation pain and vasculitis to patients, especially for the elderly and children with hardened or thin vascular walls.

[0003] CN110538144A uses 5-10 mmol / L trisodium phosphate solution as a dilute alkali solution to neutralize the low pH value of ornidazole injection during clinical administration, so that the pH value of ornidazole injection is maintained in the range of 5.0-6.0 during infusion, so as to alleviate the irritation of administration and the vasculitis caused. However, this technology requires the simultaneous use of two preparations in clinical practice, which increases the complexity of clinical medication.

[0004] CN107496351A, CN107041868A and CN110934824B dissolve left ornidazole in organic system to increase left ornidazole solubility, to improve the pH value of clinical infusion, and then improve infusion irritation.But the organic solvents such as ethanol and propylene glycol in organic system have certain safety risks to human body, such as propylene glycol can cause central nervous system depression, especially children and patients with renal failure.And the high concentration left ornidazole solution prepared in these technical schemes, during clinical medication, needs to be diluted in physiological saline injection or glucose injection for intravenous infusion, which will inevitably cause the concentration of organic solvent to decrease significantly, causing the solubility of left ornidazole to decrease, so it is easy to cause the problem of drug precipitation, which brings great safety hazards to clinical medication.

[0005] Levoornidazole disodium phosphate is a prodrug of levornidazole. Levoornidazole disodium phosphate is different from the twice-a-day usage and dosage of levornidazole sodium chloride injection. It is administered once a day, and the efficacy can be maintained for 24 hours, improving patient compliance with medication. In addition, the water solubility of levornidazole disodium phosphate is increased, and the pH value is about 5.5 when prepared as an intravenous preparation. Animal experiments have shown that this product will not cause phlebitis. However, the dosage form of this product is a sterile lyophilized powder for injection, which has high production process requirements, is difficult to ensure sterility, and greatly increases the safety risk of the sample. It also needs to be diluted and mixed with normal saline injection or 5% glucose injection before use, which increases the complexity of clinical medication.

[0006] In summary, the levornidazole fat emulsion injection in the prior art has poor water solubility, a high pH value, strong irritation, and potential safety hazards. Summary of the invention

[0007] In view of the deficiencies in the prior art, the present invention provides an application of a solubilizer in a levornidazole derivative fat emulsion injection, which solves the problems of the levornidazole fat emulsion injection in the prior art, such as poor water solubility, high pH acidity value, and strong irritation, thereby improving the safety and stability of the fat emulsion injection.

[0008] The technical solution adopted by the invention is as follows:

[0009] The invention discloses an application of a solubilizer in a levornidazole derivative fat emulsion injection, wherein the solubilizer is one or more selected from isopropyl myristate or isopropyl palmitate.

[0010] Furthermore, the levornidazole derivative fat emulsion injection includes injection oil, which is a mixture of medium-chain triglycerides and soybean oil, and the mass ratio of the medium-chain triglycerides to the soybean oil is 1:1.

[0011] Furthermore, the mass ratio of the medium chain triglyceride, soybean oil and solubilizer is 10:10:0.1-2.

[0012] Furthermore, the mass ratio of the medium chain triglyceride, soybean oil and solubilizer is 10:10:0.5-1.

[0013] Furthermore, the levornidazole derivative fat emulsion injection comprises an emulsifier, and the emulsifier is selected from one or more of egg yolk lecithin PL-100M or egg yolk lecithin E80.

[0014] Furthermore, the preparation method of the levornidazole derivative fat emulsion injection comprises the following steps:

[0015] S1. Weigh the injection oil, solubilizer and fat-soluble stabilizer or weigh the injection oil and solubilizer, mix them evenly, heat to 50-80° C., add the emulsifier, stir or high-speed shear to dissolve it, then add the levonidazole derivative, stir and dissolve it with nitrogen filling to obtain the oil phase;

[0016] S2. Disperse the isotonicity regulator in an appropriate amount of water for injection, and heat to 50-80° C. to obtain an aqueous phase;

[0017] S3, mixing the water phase described in S2 and the oil phase described in S1 at 50-80° C. under nitrogen protection, and high-speed shear emulsification for 5-15 minutes to obtain colostrum;

[0018] S4, high-pressure homogenizing the colostrum obtained in S3, making up the volume with water for injection, and adjusting the pH to the target pH with a pH adjuster to obtain final milk;

[0019] S5, nitrogen replacement of the final milk obtained in S4;

[0020] S6. Filter, fill, nitrogen-fill, seal and sterilize the final milk after nitrogen replacement to obtain a levornidazole derivative fat emulsion injection.

[0021] Furthermore, after filling, nitrogen is charged to control the residual oxygen content in the headspace to be less than or equal to 2%.

[0022] Furthermore, the dissolved oxygen concentration of the oil phase and the final milk is less than or equal to 0.8 ppm.

[0023] Furthermore, the pressure of the high-pressure homogenization is 5000-20000 psi, and the colostrum obtained in S3 is subjected to high-pressure homogenization 3-5 times;

[0024] In a preferred embodiment, the high-pressure homogenization pressure is 10,000 to 18,000 psi, and the colostrum obtained in S3 is subjected to high-pressure homogenization for 3 to 5 times.

[0025] Furthermore, the sterilization includes: using high-pressure steam sterilization, the sterilization temperature is 100-121° C., and the sterilization time is 12-45 minutes.

[0026] Furthermore, the fat-soluble stabilizer is selected from one or more of oleic acid, sodium oleate, and Kolliphor HS15.

[0027] Furthermore, the isotonicity adjusting agent is selected from one or more of sucrose, glucose, sorbitol, xylitol, sodium chloride and glycerol.

[0028] Further, the pH regulator is selected from one or more of citric acid, hydrochloric acid, fumaric acid, lysine, tartaric acid, histidine, sodium citrate, sodium hydroxide, sodium dihydrogen phosphate, and disodium hydrogen phosphate, and the pH is adjusted to 5.0-8.0;

[0029] In a preferred embodiment, the pH adjuster is selected from one or more of citric acid, hydrochloric acid, fumaric acid, lysine, tartaric acid, histidine, sodium citrate, sodium hydroxide, sodium dihydrogen phosphate, and disodium hydrogen phosphate, and the pH is adjusted to 6.0-7.0.

[0030] Furthermore, the filtration described in S6 is to reduce bacteria using a single or multiple filter membranes with pore sizes of 0.22 μm, 0.45 μm, 0.8 μm, and 1.2 μm;

[0031] Furthermore, the filter membrane used for filtration is made of PES, PVDF or PTFE.

[0032] Beneficial Effects

[0033] 1. The solubilizer of the present invention is selected from isopropyl myristate or isopropyl palmitate or a combination of the two, and is compounded with the injection oil in a certain proportion to significantly improve the solubility of the levornidazole derivative, and the solubility is increased from 0.5g / g-0.6g / g to 1.0-1.1g / g, which is increased by 2 times, thereby increasing the drug loading, avoiding the use of additional organic solvents, making the pH of the prepared levornidazole derivative fat emulsion injection close to neutral, consistent with the human body's tolerance pH, reducing the irritation generated during injection, and making clinical use more convenient.

[0034] 2. The solubilizer used in the present invention was previously mainly used in external preparations, but in the present invention, the effect of combining with the oil phase is very significant, which significantly improves the solubility of the drug in the oil phase, avoids drug precipitation, increases the stability of the levonidazole derivative fat emulsion injection, and enables the fat emulsion to be stored for a long time.

[0035] 3. The present invention also controls the dissolved oxygen concentration of the solution during the preparation process and the residual oxygen content in the headspace after filling, thereby reducing the decomposition of the main drug and the generation of oxidative impurities during storage, and improving the storage stability of the levonidazole derivative fat emulsion injection. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a particle size diagram of the levornidazole derivative fat emulsion injection of the present invention.

[0037] Figure 2 This is a chromatogram of related substances of the levornidazole derivative fat emulsion injection in Example 1 of the present invention.

[0038] Figure 3 It is a chromatogram of related substances of the levornidazole derivative fat emulsion injection in Comparative Example 5 of the present invention.

[0039] Figure 4 This is the white, opalescent spectrum of the 12-month levonidazole derivative fat emulsion injection in Example 1 of the present invention.

[0040] Figure 5 This is the white, bleached oil spectrum of the 12-month levonidazole derivative fat emulsion injection in Example 3 of the present invention. DETAILED DESCRIPTION

[0041] The present invention is further described in detail below in conjunction with specific examples and drawings. The following examples are only illustrative and not restrictive, and the protection scope of the present invention cannot be limited thereto. Unless otherwise specified, the raw materials used can be obtained from the market or homemade.

[0042] The impurities and content of the levonidazole derivative fat emulsion injection obtained in the examples and comparative examples of the present invention are determined using a high performance liquid chromatograph; the particle size, large emulsion particles larger than 5 μm, free fatty acids, peroxide value and anisidine value are detected by referring to the specific methods recommended in Part II of the 2020 edition of the "Chinese Pharmacopoeia" - Fat Emulsion Injection (C14-24); the dissolved oxygen concentration is measured by the dissolved oxygen probe provided in the liquid preparation tank; the headspace residual oxygen content is detected by a headspace residual oxygen meter / dissolved oxygen multifunctional analyzer (OXYTouch, Shanghai Qiyi Instrument). The data such as the average particle size are analyzed, and the typical particle size spectrum is shown in Figure 1 .

[0043] Test Example 1: Selection of the type and amount of injection oil

[0044] The preparation of drug-loaded fat emulsion requires that the main drug has a solubility in the injection oil. Otherwise, a stable drug-loaded fat emulsion cannot be prepared. The present invention uses a solubilizing agent in the levornidazole derivative fat emulsion injection, so that the prepared levornidazole derivative fat emulsion injection can be stored for a long time and should be stable for at least 1 year.

[0045] The solubility of left-ornidazole derivative N1 in medium-chain triglycerides (MCT), sesame oil, soybean oil, castor oil and peanut oil has been investigated in the patent CN115531306A. The solubility of left-ornidazole derivative N1 in MCT is 0.5-0.6g / g, the solubility in soybean oil is 0.3-0.4g / g, and the solubility in sesame oil, castor oil and peanut oil is less than 0.3g / g. Although the compound has the highest solubility in MCT, pure medium-chain fat emulsion will have transient high-chain fatty acid hyperemia due to its rapid hydrolysis rate in vivo, which may cause neurotoxicity, chest tightness, sweating and other discomfort symptoms, and direct infusion into the human body has high side effects. At present, most of the drug-loaded fat emulsions widely used in clinical practice still use soybean oil as the oil phase of long-chain fat emulsions or medium / long-chain fat emulsions composed of 50% medium-chain triglycerides and 50% soybean oil. Long-chain fat emulsion injection is rich in essential fatty acids such as linoleic acid and linolenic acid, among which linoleic acid is the main one. Due to its high linoleic acid content, it can affect granulocyte activity and thus immune function. The metabolism of long-chain fat emulsions and entry into mitochondria requires carnitine transport, and the oxidative metabolism rate is slow. Long-term use can easily lead to fat deposition and hypertriglyceridemia. Compared with traditional long-chain fat emulsions, medium / long-chain fat emulsions have faster metabolism and energy supply, and a lower risk of liver damage. The present invention comprehensively considers the safety of clinical use and the solubility of the drug in the injection oil. The injection oil used in the levonidazole derivative fat emulsion is a mixed oil of 50% medium-chain triglycerides and 50% soybean oil in equal proportions.

[0046] The present invention has more carefully investigated the dissolution of the levornidazole derivative in a mixture of medium-chain triglycerides and soybean oil in equal proportions. The solubility of the levornidazole derivative N1 in MCT:soybean oil with a weight ratio of 1:1 is about 0.5 g / g to 0.6 g / g, which is higher than the solubility of the levornidazole derivative N1 in soybean oil. At the same time, the present invention has creatively found that the solubility of the levornidazole derivative N1 is significantly improved by adding isopropyl myristate or isopropyl palmitate to the injection oil.

[0047] Detailed experimental operation:

[0048] Weigh an appropriate amount of left-ornidazole derivative N1, place it in a mixture of 50% medium-chain triglycerides (MCT) and 50% soybean oil heated to 70°C, so that the concentration of left-ornidazole derivative N1 is 0.4-0.6g / g, heat and stir, and observe the dissolution state; add different proportions of isopropyl myristate or isopropyl palmitate to the mixture of 50% medium-chain triglycerides (MCT) and 50% soybean oil, mix them, heat to 70°C, and then weigh an appropriate amount of left-ornidazole derivative N1, so that the concentration of left-ornidazole derivative N1 is 0.6g / g-1.2g / g, heat and stir, and observe the dissolution state. The dissolved drug solution is stored under refrigeration conditions of 2-8°C for 72 hours, and a low-temperature challenge test is performed to observe the changes in the appearance of the solution after 72 hours of low-temperature storage. If the solution becomes turbid, it means that the fat emulsion prepared with the oil solution of this concentration has the risk of drug precipitation during long-term storage. The scheme and results are shown in Tables 1 to 5.

[0049] Table 1 Solubility of levonidazole derivative N1 in injection oil - MCT: soybean oil (weight ratio 1:1)

[0050]

[0051] Results: After 72 h of low temperature challenge, the solubility of the levonidazole derivative N1 in MCT:soybean oil with a weight ratio of 1:1 was about 0.5 g / g to 0.6 g / g, which was higher than the solubility of the levonidazole derivative N1 in soybean oil.

[0052] Table 2 Solubility of levonidazole derivative N1 in oil for injection - MCT: soybean oil: isopropyl myristate / isopropyl palmitate (weight ratio 10:10:0.2)

[0053]

[0054]

[0055] Results: After 72 h of low temperature challenge, the solubility of the levonidazole derivative N1 in the MCT:soybean oil:isopropyl myristate / isopropyl palmitate with a weight ratio of 10:10:0.2 was about 0.6 g / g-0.8 g / g, which was higher than the solubility of the levonidazole derivative N1 in the MCT and soybean oil with a weight ratio of 1:1.

[0056] Table 3 Solubility of levonidazole derivative N1 in oil for injection - MCT: soybean oil: isopropyl myristate / isopropyl palmitate (weight ratio 10:10:1)

[0057]

[0058] Results: After 72 h of low temperature challenge, the solubility of levonidazole derivative N1 in MCT:soybean oil:isopropyl myristate / isopropyl palmitate with a weight ratio of 10:10:1 was about 0.8 g / g to 1.0 g / g.

[0059] Table 4 Solubility of levonidazole derivative N1 in oil for injection - MCT: soybean oil: isopropyl myristate / isopropyl palmitate (weight ratio 10:10:2)

[0060]

[0061] Results: After 72 h of low temperature challenge, the solubility of levonidazole derivative N1 in MCT:soybean oil:isopropyl myristate / isopropyl palmitate with a weight ratio of 10:10:2 was about 1.0-1.1 g / g.

[0062] Table 5 Solubility of levonidazole derivative N1 in oil for injection - MCT: soybean oil: isopropyl myristate / isopropyl palmitate (weight ratio 10:10:3)

[0063]

[0064] Results: After 72 h of low temperature challenge, the solubility of the levonidazole derivative N1 in MCT:soybean oil:isopropyl myristate / isopropyl palmitate with a weight ratio of 10:10:3 was still around 1.0-1.1 g / g.

[0065] Conclusion: The solubility of levornidazole derivative N1 was significantly improved by adding isopropyl myristate or isopropyl palmitate to the mixed oil of MCT and soybean oil in equal proportions. MCT: soybean oil: isopropyl myristate / isopropyl palmitate = 10:10:2, and the solubility was 1.0-1.1 g / g. The solubility did not increase significantly when the amount of isopropyl myristate or isopropyl palmitate was further increased. Considering the comprehensive solubility and safety, the maximum proportion of isopropyl myristate / isopropyl palmitate in fat emulsion is 2%.

[0066] The fat emulsion injection currently used in clinical practice has an injection oil content of 10-30% (w / v), of which 10%-20% w / v) is often used for direct intravenous injection. Considering the solubility, clinical dose, type of injection oil (MCT: soybean oil = 1:1) and safety of the levonidazole derivative N1, the preferred amount of injection oil is 10-20% (w / v). The solubilizers are selected from isopropyl myristate and isopropyl palmitate, which can be used alone or in combination, and the concentration in the entire fat emulsion injection is 0.1-2% (w / v), preferably 0.5-1% (w / v).

[0067] Test Example 2: Determination of drug loading

[0068] Based on the solubility and clinical safety of the left-ornidazole derivative N1, we selected MCT for injection and soybean oil for injection in equal proportions as the injection oil, and the dosage is preferably 10-20% (w / v); isopropyl myristate and / or isopropyl palmitate are used as solubilizers, and the dosage is 0.1-2% (w / v). In this solvent system, the solubility of the left-ornidazole derivative N1 is about 0.6g / g-1.0g / g, and the drug loading of the left-ornidazole derivative N1 in the left-ornidazole derivative fat emulsion is 0.1-20% (w / v), preferably 6-15% (w / v).

[0069] Test Example 3: Selection of emulsifier type and dosage

[0070] It may be related to the physical and chemical properties of the compound itself. We found that not all lecithins are suitable for the preparation of levonidazole derivative fat emulsion injection. In the experiment, we conducted parallel investigations on four injection-grade lecithins from three manufacturers, namely soybean lecithin S80 (purchased from Shanghai Taiwei Pharmaceutical Co., Ltd.), egg yolk lecithin PL-100M (purchased from Shanghai Avite Pharmaceutical Technology Co., Ltd.), egg yolk lecithin E80 (purchased from Shanghai Dongshang Industrial Co., Ltd.), and egg yolk lecithin PC-98T (purchased from Shanghai Avite Pharmaceutical Technology Co., Ltd.).

[0071] The specific prescription is shown in Table 6. The preparation method is as follows: weigh 50g of soybean oil for injection, 50g of medium-chain triglyceride for injection, 5g of isopropyl myristate and 0.3g of oleic acid, mix them, heat to 70°C, add 12g of lecithin, stir or high-speed shear to dissolve it, then add 60g of left ornidazole derivative N1, stir and dissolve it with nitrogen filling to obtain an oil phase; weigh 22g of glycerol and disperse it in 800ml of water for injection, heat to 70°C to obtain an aqueous phase; the aqueous phase and the oil phase are mixed at 70°C under nitrogen protection, and high-speed shear emulsification is performed for 10 minutes to obtain colostrum; the colostrum is subjected to high-pressure homogenization, homogenized 3 times at a homogenization pressure of 18000psi, and the volume is adjusted to 1000ml with water for injection, and the pH value is adjusted to 6.5 with hydrochloric acid to obtain the final milk; the final milk is replaced with nitrogen so that the dissolved oxygen concentration of the final milk is not greater than 0.8ppm. A levornidazole derivative fat emulsion injection is obtained by filtering with a 0.45 μm PES filter membrane, filling, nitrogen filling (controlling the headspace residual oxygen content of the drug solution after filling to be no higher than 2%), sealing, and sterilizing at 121° C. for 15 minutes.

[0072] Table 6 Prescription of levonidazole derivative fat emulsion injection prepared from different lecithin (1000ml)

[0073]

[0074] The quality of levornidazole derivative fat emulsion injection prepared with four injection-grade lecithins was evaluated after sterilization. The results are shown in Table 7.

[0075] Table 7 Quality evaluation of levonidazole derivative fat emulsion injection prepared from different types of lecithin

[0076]

[0077] Results: The experiment showed that lecithin S80 and PC-98T were not suitable as emulsifiers for left-ornidazole derivative fat emulsions. For example, in prescriptions 1 and 4, there were problems of floating oil on the liquid surface and significant increase in particle size after sterilization, especially when the large emulsion particles of 5 μm or more exceeded 0.05%. When the large emulsion particles of 5 μm or more> 0.05%, it would cause safety risks such as pulmonary embolism and alveolar tissue damage, which is unacceptable for fat emulsions. The left-ornidazole derivative fat emulsion injection prepared with egg yolk lecithin PL-100M and egg yolk lecithin E80 met the requirements in appearance, particle size and large emulsion particles of 5 μm or more. Therefore, egg yolk lecithin was selected as the emulsifier for the left-ornidazole derivative fat emulsion injection of the present invention, preferably egg yolk lecithin PL-100M and egg yolk lecithin E80. It can be used alone or in combination, and when the dosage is 0.5-10% (w / v), a safe and stable left-ornidazole derivative fat emulsion injection can be obtained.

[0078] Test Example 4: A method for preparing a levonidazole derivative fat emulsion injection

[0079] The present invention provides a method for preparing a levornidazole derivative fat emulsion injection, wherein the proportions of the components are as shown in the following table: (the following percentages refer to w / v (mass / volume))

[0080] Components Mass / Volume Levo-ornidazole derivative N1 0.1～20％ Injection oil 5～30％ Solubilizer 0.1～2％ Lecithin 0.5～10％ Stabilizer 0～0.03％ glycerin 2.2％ pH Adjusters Adjust pH to 5.0-8.0 Water for injection Add to 100%

[0081] The main drug is a levonidazole derivative N1, which is a compound shown in the structural formula, including its racemate, stereoisomer, pharmaceutically acceptable salt or solvate, or a solvate of a pharmaceutically acceptable salt:

[0082]

[0083] Test Example 4 includes 9 levornidazole derivative fat emulsion injections, which are Examples 1 to 4 and Comparative Examples 1 to 5. The prescriptions are shown in Table 8 and Table 9, respectively.

[0084] Table 8 Prescription of left ornidazole derivative fat emulsion injection

[0085]

[0086]

[0087] Table 9 Prescription of left ornidazole derivative fat emulsion injection

[0088]

[0089] Example 1 Preparation method: Weigh soybean oil for injection, medium-chain triglycerides for injection and isopropyl myristate, mix them evenly, heat them to 70°C, add lecithin, stir or high-speed shear to dissolve it, then add the left-ornidazole derivative N1, stir and dissolve it under nitrogen to obtain an oil phase; weigh glycerol and disperse it in 800 ml of water for injection, heat it to 70°C to obtain an aqueous phase; mix the aqueous phase and the oil phase at 70°C under nitrogen protection, and emulsify them at high speed shear for 10 minutes to obtain colostrum; and high-pressure homogenize the colostrum.

[0090] Preparation method of Examples 2 to 4: Weigh soybean oil for injection, medium-chain triglycerides for injection, isopropyl myristate and / or isopropyl palmitate and oleic acid, mix well, heat to 70°C, add lecithin, stir or high-speed shear to dissolve it, then add left-ornidazole derivative N1, stir and dissolve under nitrogen to obtain an oil phase; weigh glycerol and disperse it in 800 ml of water for injection, heat to 70°C to obtain an aqueous phase; mix the aqueous phase and the oil phase at 70°C under nitrogen protection, emulsify at high speed shear for 10 minutes to obtain colostrum; and high-pressure homogenize the colostrum.

[0091] Example 1 Homogenize 5 times under a homogenizing pressure of 10000psi, dilute to 1000ml with water for injection, adjust pH value to 5.01 with hydrochloric acid to obtain final milk; replace nitrogen with the final milk to make the dissolved oxygen concentration of the final milk not more than 0.8ppm. Filter with a 0.8μm PTFE filter membrane, fill, nitrogen fill (control the headspace residual oxygen content of the liquid after filling not more than 2%), seal, and sterilize at 100℃ for 45min to obtain a levonidazole derivative fat emulsion injection.

[0092] Example 2 Homogenize 3 times under 18000psi homogenization pressure, dilute to 1000ml with injection water, adjust pH value to 5.99 with citric acid to obtain final milk; nitrogen replace the final milk to make the dissolved oxygen concentration of the final milk not more than 0.8ppm. Filter with 0.22μm PVDF filter membrane, fill, nitrogen fill (control the headspace residual oxygen content of the liquid after filling not more than 2%), seal, sterilize at 121℃ for 12min, and obtain a levonidazole derivative fat emulsion injection.

[0093] Example 3 Homogenize 3 times at 20000psi homogenization pressure, dilute to 1000ml with water for injection, adjust pH to 7.03 with sodium dihydrogen phosphate and disodium hydrogen phosphate to obtain final milk; nitrogen replace the final milk to make the dissolved oxygen concentration of the final milk not more than 0.8ppm. Filter with 1.2μm PES filter membrane, fill, nitrogen fill (control the headspace residual oxygen content of the liquid after filling not more than 2%), seal, sterilize at 117℃ for 30min, and obtain a levonidazole derivative fat emulsion injection.

[0094] Example 4 Homogenize 3 times at 20000psi homogenization pressure, dilute to 1000ml with injection water, adjust to 7.93 with sodium citrate to obtain final milk; nitrogen replace the final milk to make the dissolved oxygen concentration of the final milk not more than 0.8ppm. Filter with 1.2μm PES material filter membrane, fill, nitrogen fill (control the headspace residual oxygen content of the liquid after filling is not higher than 2%), seal, and sterilize at 121℃ for 15min to obtain a levonidazole derivative fat emulsion injection.

[0095] Comparative Examples 1 to 4: The formulation is slightly different from that of Examples 1 to 4 (without a solubilizer, and using emulsifiers such as poloxamer 188, polysorbate 80, and ethanol). The preparation method is as follows: weigh soybean oil for injection and / or medium-chain triglycerides for injection, an emulsifier (poloxamer 188, polysorbate 80 or ethanol) and oleic acid, mix well, heat to 70° C., add lecithin, stir or high-speed shear to dissolve it, then add the left ornidazole derivative N1, and stir with nitrogen. Dissolve to obtain the oil phase; weigh glycerol and disperse it in 800 ml of water for injection, heat to 70°C to obtain the water phase; mix the water phase and the oil phase at 70°C under nitrogen protection, and emulsify at high speed for 10 minutes to obtain colostrum; homogenize 3 times under a homogenizing pressure of 18000psi, make the volume to 1000 ml with water for injection, adjust the pH value to about 6.0 with hydrochloric acid to obtain the final milk; replace the final milk with nitrogen so that the dissolved oxygen concentration of the final milk is not more than 0.8ppm.

[0096] The product is filtered through a 0.22 μm PVDF filter membrane, filled, nitrogen-filled (the residual oxygen content in the headspace of the drug solution after filling is controlled to be no higher than 2%), sealed, and sterilized at 121° C. for 12 minutes to obtain a levonidazole derivative fat emulsion injection.

[0097] Comparative Example 5: A levonidazole derivative fat emulsion injection

[0098] The prescription is exactly the same as that in Example 2. The difference between the preparation method and Example 2 is that the preparation process is not nitrogen-filled and the liquid medicine is nitrogen-filled before and after filling to control the residual oxygen content in the headspace to about 3%. The specific operation is: weigh soybean oil for injection, medium-chain triglycerides for injection and oleic acid, mix them, heat to 70°C, add lecithin, stir or high-speed shear to dissolve them, then add left ornidazole derivative N3, stir and dissolve with nitrogen to obtain oil phase; weigh glycerol and disperse it in 800ml water for injection, heat to 70°C to obtain water phase; mix the water phase and oil phase at 70°C without nitrogen protection, emulsify at high speed shear for 10 minutes to obtain colostrum; and high-pressure homogenize the colostrum. Homogenize 5 times at a homogenizing pressure of 10000psi, make the volume to 1000ml with water for injection, adjust the pH value to 5.97 with hydrochloric acid to obtain a final emulsion; directly filter with a 0.8μm PTFE filter membrane, fill, nitrogen fill (control the headspace residual oxygen content of the drug solution after filling to about 3%), seal, and sterilize at 100℃ for 45min to obtain a levonidazole derivative fat emulsion injection.

[0099] Test Example 5: Quality evaluation of the levonidazole derivative fat emulsion injection of the present invention

[0100] The quality of the levornidazole derivative fat emulsion injection prepared in Examples 1 to 4 and Comparative Examples 1 to 5 was evaluated. The main evaluation items were: appearance, pH value, particle size, large emulsion particles larger than 5 μm, impurities, peroxide value and anisidine value. Specific data are shown in Tables 10 and 11.

[0101] Table 10 Quality evaluation of left ornidazole derivative fat emulsion injection

[0102]

[0103] Table 11 Quality evaluation of levonidazole derivative fat emulsion injection

[0104]

[0105]

[0106] Conclusion: The quality attributes of the samples obtained at 0 time of Examples 1 to 4 and Comparative Example 5 all meet the quality requirements of the present invention, but the impurities, peroxide value and anisidine value of Comparative Example 5 are significantly higher than those of Examples 1 to 4. The reason for this is that the nitrogen protection during the preparation process and after filling of Comparative Example 5 is not strong enough, and the unsaturated fatty acids in the oil and phospholipids contained in the fat emulsion are oxidized to form peroxides during the high-temperature sterilization process, which further decompose to form aldehydes or ketones. The appearance and particle size of the samples at 0 time of Comparative Examples 1 to 4 are both worse than those of Examples 1 to 4. The reason for this is that there is no solubilizer (isopropyl myristate or isopropyl palmitate) in the prescription, and the solubility of the active ingredient levonidazole derivative N1 in oil is not enough. Even if an emulsifier (poloxamer 188, polysorbate 80, ethanol) is added, it cannot be prepared into a fat emulsion injection with a concentration of 15% (w / v).

[0107] Test Example 6: Investigation on the long-term stability of the levornidazole derivative fat emulsion injection of the present invention.

[0108] The samples prepared in Examples 1 to 4 and Comparative Example 5 were placed at 25°C ± 2°C and a humidity of 60% ± 5%. Samples were taken in October, March, June, September and December, respectively, and the changes in appearance, pH value, particle size, large milk particles larger than 5 μm, impurities, peroxide value and anisidine value were mainly investigated. The results are shown in Table 12. Example 1 and Comparative Example 5 were placed at 25°C ± 2°C and a humidity of 60% ± 5% for 12 months. The chromatograms of the relevant substances are shown in Table 12. Figure 2 and Figure 3 .

[0109] Table 12 Long-term stability data of levonidazole derivative fat emulsion injection

[0110]

[0111]

[0112] Conclusion: The left ornidazole derivative fat emulsion injection (Examples 1 to 4) disclosed in the present invention, which controls dissolved oxygen during the preparation process and controls residual oxygen in the headspace by nitrogen filling after filling, is placed at 25°C ± 2°C and 60% ± 5% humidity for 6 months, and its appearance, pH value, particle size, large emulsion particles larger than 5μm, impurities, peroxide value and anisidine value do not show significant changes. However, after continued placement, the particle size and appearance of Examples 3 and 4 have undergone qualitative changes in 9 months. Example 3 has a tendency to demulsify in 9 months and completely demulsifies in 12 months; Example 4 has completely demulsified in 9 months. In comparison, Examples 1 and 2 are placed at room temperature for 12 months, and the physical and chemical properties such as appearance, particle size and impurities remain stable. The upper limit of the drug loading of the left ornidazole derivative fat emulsion involved in the present invention is preferably 15% (w / v).

[0113] In addition, in Comparative Example 5, the dissolved oxygen was not controlled during the preparation process and the headspace residual oxygen level was not controlled to be higher than 2% by nitrogen filling after filling. Under the same storage conditions, its appearance, pH value, particle size, large emulsion particles larger than 5 μm, impurities, peroxide value and anisidine value all changed significantly, proving that its stability was far worse than that of Examples 1 to 4. It is shown that it is very necessary to control the dissolved oxygen in the preparation process and the headspace residual oxygen by nitrogen filling after filling of a levornidazole derivative fat emulsion injection involved in the present invention, which can reduce the oxidation of oil and phospholipids in the fat emulsion during the preparation, sterilization and storage process, improve the stability and safety of the fat emulsion, and ensure the quality of the fat emulsion.

[0114] The above results show that the levornidazole derivative fat emulsion injection involved in the present invention significantly improves the drug loading and stability compared with the prior art, can be stored for a long time, and has more stable quality.

Claims

1. A solubilizing agent for use in a levornidazole derivative fat emulsion injection, characterized in that: The solubilizing agent is one or more selected from isopropyl myristate or isopropyl palmitate.

2. The use of the solubilizing agent according to claim 1 in a levornidazole derivative fat emulsion injection, characterized in that: The levornidazole derivative fat emulsion injection comprises injection oil, which is a mixture of medium-chain triglycerides and soybean oil, and the mass ratio of the medium-chain triglycerides to the soybean oil is 1:

1.

3. Use of the solubilizing agent according to claim 2 in a levornidazole derivative fat emulsion injection, characterized in that: The mass ratio of the medium chain triglyceride, soybean oil and solubilizer is 10:10:0.1-2.

4. Use of the solubilizing agent according to claim 3 in levornidazole derivative fat emulsion injection, characterized in that: The mass ratio of the medium chain triglyceride, soybean oil and solubilizer is 10:10:0.5-1.

5. Use of the solubilizing agent according to claim 4 in levornidazole derivative fat emulsion injection, characterized in that: The levornidazole derivative fat emulsion injection comprises an emulsifier, and the emulsifier is selected from one or more of egg yolk lecithin PL-100M and egg yolk lecithin E80.

6. Use of the solubilizing agent according to claim 5 in a levornidazole derivative fat emulsion injection, characterized in that: The preparation method of the levornidazole derivative fat emulsion injection comprises the following steps: S1. Weigh the injection oil, solubilizer and fat-soluble stabilizer or weigh the injection oil and solubilizer, mix them evenly, heat to 50-80° C., add the emulsifier, stir or high-speed shear to dissolve it, then add the levonidazole derivative, stir and dissolve it with nitrogen filling to obtain the oil phase; S2, dispersing the isotonicity adjusting agent in water for injection, heating to 50-80°C to obtain an aqueous phase; S3, mixing the water phase described in S2 and the oil phase described in S1 at 50-80° C. under nitrogen protection, and high-speed shear emulsification for 5-15 minutes to obtain colostrum; S4, high-pressure homogenizing the colostrum obtained in S3, making up the volume with water for injection, and adjusting the pH to the target pH with a pH adjuster to obtain final milk; S5, nitrogen replacement of the final milk obtained in S4; S6. Filter, fill, nitrogen-fill, seal and sterilize the final milk after nitrogen replacement to obtain a levornidazole derivative fat emulsion injection.

7. Use of the solubilizing agent according to claim 6 in a levornidazole derivative fat emulsion injection, characterized in that: After filling, nitrogen is added to control the residual oxygen content in the headspace to be less than or equal to 2%.

8. Use of the solubilizing agent according to claim 6 in a levornidazole derivative fat emulsion injection, characterized in that: The dissolved oxygen concentration of the oil phase and the final milk is less than or equal to 0.8 ppm.

9. Use of the solubilizing agent according to claim 6 in a levornidazole derivative fat emulsion injection, characterized in that: The pressure of the high-pressure homogenization is 5000-20000 psi, and the colostrum obtained in S3 is subjected to high-pressure homogenization for 3-5 times.

10. Use of the solubilizing agent according to claim 6 in a levornidazole derivative fat emulsion injection, characterized in that: The sterilization includes: using high-pressure steam sterilization, the sterilization temperature is 100-121° C., and the sterilization time is 12-45 minutes.

Images