A letimovir injection and a preparation method thereof

Abstract

The present application relates to the technical field of pharmaceutical preparation, and specifically discloses a letemovir injection and a preparation method thereof. A prescription amount of letemovir, hydroxypropyl-beta-cyclodextrin and sodium hydroxide are directly mixed into a dry powder once, and then added into water for injection with a temperature of 40-70 DEG C for stirring and dissolving, filtering, filling, sterilizing, and obtaining the letemovir injection. The preparation method of the letemovir injection provided by the present application effectively reduces the difficulty in preparing the letemovir injection, avoids the generation of bubbles, wall sticking, opalescence and precipitation in the preparation process, is simple to operate, does not require special equipment, has high industrial feasibility, and has high quality stability of the prepared letemovir injection, with a total impurity content of ≤0.15% in 12 months, a significantly reduced increase in impurity content during storage, always being a colorless and clear solution in property, and a stable pH value, which is conducive to improving the clinical application safety of the letemovir injection and can realize effective substitution of the original research.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical preparations, in particular to a letimovir injection and a preparation method thereof. BACKGROUND

[0002] The letimovir injection is a new non-nucleoside cytomegalovirus (CMV) inhibitor developed by Merck Company. It effectively prevents the processing and packaging of viral DNA by targeting the viral endonuclease complex, thereby exerting an antiviral effect. Letimovir has high selectivity and strong effect on CMV, and has higher selectivity and strength compared with DNA polymerase inhibitors. Clinical studies have shown that letimovir can effectively prevent CMV infection in patients, and has no obvious adverse reactions such as bone marrow suppression, increased nephrotoxicity, and no cross-resistance restrictions. The two dosage forms of letimovir (tablets and injections) can be used interchangeably, and no dose adjustment is required when changing, providing personalized prevention programs for the clinic. Letimovir injection is suitable for patients who cannot take oral drugs, especially for adult recipients who are CMV seropositive after allogeneic hematopoietic stem cell transplantation (allo-HSCT), for the prevention of CMV infection and CMV disease. Letimovir injection has been approved for marketing in China in May 2022. Prior to this, letimovir tablets have been approved in China in January 2022.

[0003] According to the instructions of the original reference preparation, letimovir injection contains hydroxypropyl-β-cyclodextrin (HP-β-CD) as a cosolvent, and sodium hydroxide is also added to adjust the pH value. The conventional preparation method of cyclodextrin inclusion compound generally has precipitation method and co-precipitation method. Since letimovir is almost insoluble in water, it is easy to float on the surface of the liquid, and stable bubbles will be produced during stirring, and at the same time, opalescence is easy to produce, therefore, the preparation of letimovir injection is difficult. The use of sodium hydroxide to assist dissolution can reduce the difficulty of preparation, but the particle size peak of the prepared sample is inconsistent with the original reference, which affects the inclusion effect of HP-β-CD, and may cause differences in quality and clinical efficacy, therefore, it is difficult to prepare letimovir injection with consistent quality as the original reference by using the conventional method. SUMMARY

[0004] Therefore, the present application provides a letimovir injection and a preparation method thereof. The present application solves the problems of easy production of bubbles, opalescence, and inconsistent particle size with the original reference in the preparation process of letimovir injection by directly dry mixing letimovir, hydroxypropyl-β-cyclodextrin and sodium hydroxide to obtain a mixed powder, and adding the mixed powder into water for injection, and the stability of the injection is higher, which can realize effective substitution of the original reference preparation.

[0005] To achieve the above-mentioned purposes, the technical scheme provided by the present application is as follows:

[0006] The application provides a preparation method of letimovir injection, and comprises the following steps:

[0007] In step a, letimovir, hydroxypropyl-beta-cyclodextrin and sodium hydroxide are weighed according to a prescription amount, and are uniformly mixed to obtain a mixed powder.

[0008] In step b, the mixed powder is added into part of water for injection, is dissolved, and water for injection is added to the total preparation amount to obtain letimovir liquid.

[0009] In step c, the letimovir liquid is filtered through a multi-stage polyether sulfone filter element with a pore size decreasing in turn, is filled, is sealed, is sterilized, and letimovir injection is obtained.

[0010] In step b, the temperature of the water for injection is 40-70 DEG C.

[0011] The inventor accidentally found that, by stirring and dissolving the prescription amount of letimovir, hydroxypropyl-beta-cyclodextrin and sodium hydroxide directly in water for injection with a temperature of 40-70 DEG C, letimovir can easily dissolve under the liquid surface, cannot adhere to the preparation container on the liquid surface, and no bubbles are generated during stirring.

[0012] Further, in step a, the uniform mixing is performed by using a dry mixing machine.

[0013] The application adopts a one-step dry mixing method and a one-step feeding method, greatly improves production efficiency, simplifies a process flow, and is convenient for realizing industrialized scale production application.

[0014] For example, in step a, the stirring speed of the dry mixing machine is 100-200 r / min, and the mixing time is 15-30 min.

[0015] Further, in step b, the part of water for injection is 40-70% of the total preparation amount.

[0016] After the one-step dry mixing of the materials, letimovir can directly enter the liquid surface and is completely dissolved, no insoluble particles are generated in the prepared product, no opalescence is generated, and the product quality is consistent with that of the original research, so that a simple and efficient new method is provided for letimovir injection, and the method has important significance for expanding the application of letimovir.

[0017] Further, the multi-stage filter core filtering is two-stage polyether sulfone filter core filtering, wherein the pore size of the first-stage polyether sulfone filter core is 0.45 µm, and the pore size of the second-stage polyether sulfone filter core is 0.2 µm.

[0018] The application adopts the multi-stage polyether sulfone filter core with a specific pore size for sequential filtering, thereby ensuring that the bacterial and endotoxin content of the product is qualified; in the production process, no activated carbon is used, thereby reducing the pollution to the clean production area, reducing the environmental protection pressure, reducing the impurities and insoluble particles possibly introduced by the activated carbon, and being beneficial to further improving the effectiveness, safety and stability of the product quality.

[0019] Further, in step c, the sterilization is sterilization at 121 ℃ for 8 min to 15 min.

[0020] The application adopts the terminal sterilization method, thereby improving the sterile level and sterilization efficiency of the product.

[0021] The preparation method of the letromovir injection provided by the application has simple process operation, only needs to control the mixing mode of the materials, and can prepare the letromovir injection with the same quality as the original research product by one-time feeding; meanwhile, the application adopts the terminal sterilization method and does not use activated carbon in the preparation process, thereby greatly improving the safety of the product for clinical medication, reducing the production cost, and having a wide application prospect.

[0022] The application further provides a letromovir injection prepared by the preparation method of the letromovir injection according to any one of the above.

[0023] Further, 1 mL of the letromovir injection includes: letromovir 20 mg, hydroxypropyl-β-cyclodextrin 145 mg to 155 mg, sodium hydroxide 1 mg to 1.5 mg, and sodium chloride 3 mg to 3.5 mg.

[0024] Preferably, 1 mL of the letromovir injection includes: letromovir 20 mg, hydroxypropyl-β-cyclodextrin 140 mg, sodium hydroxide 1.2 mg, and sodium chloride 3.1 mg.

[0025] Further, the specification of the letromovir injection is 12 mL: 240 mg, and 24 mL: 480 mg.

[0026] The preparation method of the letimovir injection provided by the application effectively reduces the preparation difficulty of the letimovir injection, is simple to operate, does not require special equipment, has high industrial feasibility, and has high quality stability of the prepared letimovir injection, a total impurity content of the letimovir injection prepared for 12 months is less than or equal to 0.15%, the increase of the impurity content during storage is obviously reduced, the property is always a colorless and clear solution, the pH value is stable, the clinical application safety of the letimovir injection is improved, the original research can be effectively replaced, and the letimovir injection has a wide application prospect. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical scheme and advantages of the application more clear and understandable, the application is further described in detail below in combination with examples. It should be understood that the specific examples described herein are only used to explain the application, and are not used to limit the application.

[0028] In order to better illustrate the application, the following examples are further illustrated.

[0029] Example 1

[0030] The example provides a letimovir injection, and the prescription amount is as follows:

[0031]

[0032] Water for injection is added to 1 mL.

[0033] The specific preparation steps are as follows:

[0034] Step a, letimovir, hydroxypropyl-beta-cyclodextrin, sodium hydroxide and sodium chloride are weighed according to the preparation amount, added into a dry powder mixer, stirred and mixed at 180 r / min for 20 min, and a mixed powder is obtained;

[0035] Step b, the above mixed powder is added into 40℃ water for injection with a total preparation amount of 40%, stirred and dissolved for 40 min, cooled to room temperature, and water for injection is added to the total preparation amount, and a letimovir liquid is obtained;

[0036] Step c, the letimovir liquid is filtered through a two-stage polyether sulfone filter element, wherein the pore size of the first-stage polyether sulfone filter element is 0.45 μm, and the pore size of the second-stage polyether sulfone filter element is 0.2 μm, then filled, sealed, sterilized at 121℃ for 10 min, and a letimovir injection is obtained.

[0037] The prepared letimovir injection is detected for particle size, scanning shows two nanoparticle size peaks, peak positions: 1.73 nm and 401 nm. No precipitate or white point is precipitated after low-temperature (2-8℃) storage for 10 days.

[0038] The original research (Merk Wharp, batch number: W029059) reference preparation scanning shows two nanoparticle size peaks, peak position: 1.78 nm and 415 nm. Small white point precipitates after low temperature (2-8℃) for 5 days.

[0039] Example 2

[0040] This embodiment provides a letimovir injection, the prescription amount is shown as follows:

[0041]

[0042] Water for injection to 1 mL.

[0043] The specific preparation steps are as follows:

[0044] Step a, according to the preparation amount, letimovir, hydroxypropyl-β-cyclodextrin, sodium hydroxide and sodium chloride are weighed and added into a dry powder mixer, stirred and mixed at 100 r / min for 30 min, and a mixed powder is obtained;

[0045] Step b, the above mixed powder is added into 55℃ water for injection with a total preparation amount of 50%, stirred and dissolved for 30 min, cooled to room temperature, and water for injection is added to the total preparation amount to obtain letimovir liquid;

[0046] Step c, the letimovir liquid is filtered through a two-stage polyether sulfone filter element, wherein the pore size of the first-stage polyether sulfone filter element is 0.45μm, and the pore size of the second-stage polyether sulfone filter element is 0.2μm, then filled, sealed, and sterilized at 121℃ for 12 min to obtain letimovir injection.

[0047] The prepared letimovir injection is detected for particle size, scanning shows two nanoparticle size peaks, peak position: 1.68 nm and 410 nm. No precipitate or white point precipitates after low temperature (2-8℃) for 10 days.

[0048] Example 3

[0049] This embodiment provides a letimovir injection, the prescription amount is shown as follows:

[0050]

[0051] Water for injection to 1 mL.

[0052] The specific preparation steps are as follows:

[0053] Step a, according to the preparation amount, letimovir, hydroxypropyl-β-cyclodextrin, sodium hydroxide and sodium chloride are weighed and added into a dry powder mixer, stirred and mixed at 100 r / min for 30 min, and a mixed powder is obtained;

[0054] Step b, the above mixed powder is added into 65℃ water for injection, and stirred and dissolved for 25 minutes, then cooled to room temperature, and water for injection is added to the total preparation amount to obtain the telithomycin liquid;

[0055] Step c, the telithomycin liquid is filtered through two-stage polyether sulfone filter elements, wherein the pore size of the first-stage polyether sulfone filter element is 0.45μm, and the pore size of the second-stage polyether sulfone filter element is 0.2μm, then filled, sealed, sterilized at 121℃ for 12 minutes to obtain the telithomycin injection.

[0056] The prepared telithomycin injection is detected for particle size, and scanning shows two nanoparticle size peaks, peak positions: 1.78nm and 426nm. No precipitate or white point is precipitated after being placed at low temperature (2-8℃) for 10 days.

[0057] Example 4

[0058] The present example provides a telithomycin injection, and the prescription amount is shown as follows:

[0059]

[0060] Water for injection to 1mL.

[0061] The specific preparation steps are as follows:

[0062] Step a, according to the preparation amount, telithomycin, hydroxypropyl-β-cyclodextrin, sodium hydroxide and sodium chloride are weighed and added into a dry powder mixer, stirred and mixed at 200r / min for 15 minutes to obtain a mixed powder;

[0063] Step b, the above mixed powder is added into 70℃ water for injection, and stirred and dissolved for 25 minutes, then cooled to room temperature, and water for injection is added to the total preparation amount to obtain the telithomycin liquid;

[0064] Step c, the telithomycin liquid is filtered through two-stage polyether sulfone filter elements, wherein the pore size of the first-stage polyether sulfone filter element is 0.45μm, and the pore size of the second-stage polyether sulfone filter element is 0.2μm, then filled, sealed, sterilized at 121℃ for 10 minutes to obtain the telithomycin injection.

[0065] The prepared telithomycin injection is detected for particle size, and scanning shows two nanoparticle size peaks, peak positions: 1.82nm and 396nm. No precipitate or white point is precipitated after being placed at low temperature (2-8℃) for 10 days.

[0066] The telithomycin injection samples prepared in Examples 1-4 and the reference preparation are sterilized at 121℃ for 12 minutes again, and then subjected to quality detection, and the results are shown in Table 1.

[0067] Content determination method: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2010 edition part 2 appendix V D). Chromatographic column: octadecylsilane bonded silica gel chromatographic column; mobile phase: phosphoric acid-water-methanol=3:247:750; detection wavelength: 274 nm; flow rate: 1.5 mL / min; column temperature: 30 DEG C; injection volume: 1 μL.

[0068] Related substance determination method: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2010 edition part 2 appendix V D). Chromatographic column: octadecylsilane bonded silica gel chromatographic column; mobile phase: water (adjust pH value to 2.5 by phosphoric acid)-acetonitrile=1340:680; detection wavelength: 234 nm; flow rate 2 mL / min; column temperature: 30 DEG C; injection volume: 1 μL.

[0069] Table 1

[0070]

[0071]

[0072] The results show that the properties, precipitation and opalescence and related substances of the letemovir injection prepared in the embodiments of the application are consistent with the reference preparation.

[0073] Long-term stability test

[0074] The example 1~example 3 (sterilized once) and reference preparation (batch number: W029059) are subjected to long-term stability test, temperature 25 DEG C±2 DEG C, relative humidity is 60%±10%, at 0 month, 6 months and 12 months are sampled and detected respectively, and the results are shown in table 2.

[0075] Table 2

[0076]

[0077]

[0078] The results show that the stability of the letemovir injection prepared in the embodiments of the application is good, and is consistent with the original research, and the effective substitution of the original research can be realized.

[0079] The letemovir injection prepared in example 4 can achieve the technical effects equivalent to example 1~3.

[0080] Comparative example 1

[0081] The comparative example provides a letemovir injection, which has the same formula as example 1, and the preparation method comprises the following steps:

[0082] Step a, sodium hydroxide was weighed according to the preparation amount, added to water for injection with a total amount of 45% at 55°C, stirred and dissolved to obtain a sodium hydroxide solution;

[0083] Step b, the preparation amount of hydroxypropyl-β-cyclodextrin was added to the above sodium hydroxide solution, stirred and dissolved, then the preparation amount of letromivir was added, stirred for 30 min, the preparation amount of sodium chloride was added, stirred for 35 min to dissolve, and letromivir solution was obtained;

[0084] Step c, activated carbon was added to the above letromivir injection, stirred to remove pyrogen, filtered to remove activated carbon, and filtrate was obtained; the filtrate was filtered with 0.45 μm and 0.2 μm polyether sulfone filter cartridges in turn, filled, sterilized at 121°C for 12 min, and letromivir injection was obtained.

[0085] It was found during preparation that a small part of letromivir still floated on the liquid surface and the tank wall, and the solution showed a light blue opalescence. After adding activated carbon treatment, the solution was clear, and after sterilization, there was opalescence.

[0086] Meanwhile, scanning showed that the letromivir injection prepared in the comparative example had three particle size peaks: 1.23 nm, 171 nm and 568 nm, and white spots were precipitated after standing at room temperature overnight.

[0087] Comparative Example 2

[0088] The comparative example provides a letromivir injection, which has the same formula as Example 1, and the preparation method comprises the following steps:

[0089] Step a, sodium hydroxide was weighed according to the preparation amount, added to water for injection with a total amount of 65% at 50°C, stirred and dissolved to obtain a sodium hydroxide solution;

[0090] Step b, the preparation amount of letromivir was added to the above sodium hydroxide solution, stirred for 30 min, then the preparation amount of sodium chloride was added, stirred for 35 min to dissolve, and finally the preparation amount of hydroxypropyl-β-cyclodextrin was added, stirred and dissolved to obtain letromivir solution;

[0091] Step c, activated carbon was added to the above letromivir injection, stirred to remove pyrogen, filtered to remove activated carbon, and filtrate was obtained; the filtrate was filtered with 0.45 μm and 0.2 μm polyether sulfone filter cartridges in turn, filled, sterilized at 121°C for 12 min, and letromivir injection was obtained.

[0092] It was found during preparation that a small part of letromivir still floated on the liquid surface and the tank wall, and the solution showed a light blue opalescence. After adding activated carbon treatment, the solution was clear, and after sterilization, there was opalescence.

[0093] At the same time, the scanning shows that the lovastatin injection prepared in the comparative example has three particle size peaks: 2.93 nm, 275 nm and 4560 nm, and white spots are precipitated after being placed at room temperature overnight.

[0094] Comparative Example 3

[0095] The comparative example provides a preparation method of lovastatin injection, and the specific steps are as follows:

[0096] In step a, 18.0 g of arginine is weighed and added into 160 mL of water for injection to completely dissolve, to obtain an arginine aqueous solution;

[0097] In step b, 20 g of lovastatin is dissolved in the above arginine aqueous solution, 6 g of sodium acetate trihydrate is added and stirred to dissolve; sodium hydroxide is added to adjust the pH to 5.7, and 40 mL of water for injection is added to prepare a total amount, to obtain a lovastatin liquid;

[0098] In step c, activated carbon is added to the above lovastatin liquid, and the pyrogen is removed by stirring, and the activated carbon is removed by filtration, to obtain a filtrate; the filtrate is filtered with 0.45 μm and 0.2 μm polyether sulfone filter cartridges in sequence, filled, and lovastatin injection is obtained.

[0099] The lovastatin injections prepared in Example 1 and Comparative Example 3 and the reference preparation are sterilized at 121℃ for 12 min respectively, and quality detection is performed, and the results are shown in Table 3.

[0100] Table 3

[0101]

[0102] It can be seen from the results that the impurity content of the comparative example 3 and the reference preparation obviously increases after secondary sterilization; at the same time, the total impurity of the comparative example 3 after sterilization is obviously higher than that of the reference preparation and the sample prepared in Example 1 of the application.

[0103] In summary, the lovastatin injection is prepared by the method of direct mixing and one-step feeding, which significantly shortens the preparation time, avoids the problems of air bubbles, wall sticking, opalescence and precipitation in the preparation process, is beneficial to scale-up production and improvement of product quality, improves the preparation success rate, and effectively improves the safety of clinical use, and has practical social significance and economic value.

[0104] The above only describes the preferred embodiments of the application and should not be used to limit the application, and any modification, equivalent replacement or improvement made within the spirit and principle of the application should be included in the protection scope of the application.

Claims

1. A method for preparing letermovir injection, characterized in that, Includes the following steps: Step a: Weigh out the prescribed amounts of Letermovir, hydroxypropyl-β-cyclodextrin, and sodium hydroxide, mix them thoroughly to obtain a mixed powder; Step b: Add the mixed powder to a portion of water for injection, dissolve it, and add water for injection to the total prepared amount to obtain Letermovir solution; Step c: Filter the lemetmovir solution through a multi-stage polyethersulfone filter with progressively smaller pore sizes, fill, seal, and sterilize to obtain lemetmovir injection solution; In step b, the temperature of the water for injection is 40℃~70℃; Each 1 mL of the described letermovir injection comprises: 20 mg letermovir, 145 mg to 155 mg hydroxypropyl-β-cyclodextrin, 1 mg to 1.5 mg sodium hydroxide, and 3 mg to 3.5 mg sodium chloride.

2. The method for preparing lemetmovir injection as described in claim 1, characterized in that, In step a, the uniform mixing is achieved using a dry mixer.

3. The method for preparing lemetmovir injection as described in claim 1, characterized in that, In step b, the water for injection constitutes 40% to 70% of the total prepared volume.

4. The method for preparing lemetmovir injection as described in claim 1, characterized in that, The multi-stage filter cartridge filtration is a two-stage polyethersulfone filter cartridge filtration, wherein the pore size of the first-stage polyethersulfone filter cartridge is 0.45µm and the pore size of the second-stage polyethersulfone filter cartridge is 0.2µm.

5. The method for preparing lemetmovir injection as described in claim 1, characterized in that, In step c, the sterilization is performed at 121°C for 8 to 15 minutes.

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