Process for preparation of composition comprising nystatin

Abstract

The invention provides a preparation method of a composition containing nystatin. The preparation method comprises the following steps: preparing a base solution containing a suspending aid and buffer salt; preparing a wetting solution containing nystatin; cooling the base solution to 10-15 DEG C, and adding the wetting solution to prepare a mixed solution; carrying out heat preservation on the mixed solution at 10-15 DEG C for a first time, and then carrying out heat preservation at 45-50 DEG C for a second time; and cooling the mixed solution after the heat preservation is finished, then adding a preservative and a flavoring agent, and adjusting the pH value of the mixed system to 6.5-7.2 to prepare the composition containing nystatin. The preparation method is high in production efficiency, and the prepared composition containing nystatin is stable in quality.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical formulation technology, and in particular to a method for preparing a composition containing nystatin. Background Technology

[0002] Nystatin is a polyene antibiotic (structural formula shown below). In 1950, the soil microorganism *Streptomyces noursei* was discovered to secrete an antifungal substance, thus discovering the first antifungal antibiotic. Nystatin has a conjugated polyene macrocyclic lactone structure, inhibiting the activity of fungi and dermatophytes, but has no inhibitory effect on bacteria.

[0003]

[0004] Nystatin exhibits antifungal and bactericidal effects against various yeasts and yeast-like fungi in vitro. In vitro, with increasing nystatin levels, *Candida albicans* did not show significant resistance to repeated subcultures of nystatin; however, other *Candida* species showed considerable resistance. Generally, resistance does not develop in vivo. Nystatin exerts its antifungal effect by binding to sterols in the cell membrane of susceptible *Candida*, thereby altering cell membrane permeability and causing leakage of intracellular components. Nystatin has no significant activity against bacteria, protozoa, or viruses.

[0005] Nystatin has broad-spectrum antifungal activity and high antibacterial activity against Candida spp. Cryptococcus neoformans, Aspergillus, Mucor, Microsporum, Histoplasma capsulatum, Blastomyces dermatitidis, and dermatophytes are usually also sensitive to this product.

[0006] Nystatin film-coated tablets (500,000 IU / tablet) are the only oral administration product in China that has passed the consistency evaluation for dosage form and specifications. However, for infants and young children with oral candidiasis, the safety and compliance of the medication are severely limited. Oral suspensions are more suitable for infants and young children and elderly people with swallowing difficulties, and can achieve precise drug delivery.

[0007] However, traditional oral suspension preparation processes suffer from problems such as long production time and low production efficiency. Therefore, there is an urgent need to provide a method for preparing oral suspensions with high production efficiency and stable quality. Summary of the Invention

[0008] Therefore, it is necessary to provide a method for preparing a composition containing nystatin that has high production efficiency and stable quality.

[0009] This application provides a method for preparing a composition containing nystatin, comprising the following steps:

[0010] Prepare a base solution containing suspending agents and buffer salts;

[0011] Prepare a wetting solution containing nystatin;

[0012] The base liquid is cooled to 10℃-15℃ and then the wetting liquid is added to obtain a mixed liquid.

[0013] The mixture is kept at 10℃-15℃ for a first time, and then kept at 45℃-50℃ for a second time.

[0014] After the heat preservation is completed, the mixture is cooled down, and then preservatives and flavoring agents are added. The pH value of the mixed system is adjusted to 6.5-7.2 to prepare the composition containing nystatin.

[0015] In some embodiments, the first time is greater than or equal to 2 hours.

[0016] In some embodiments, the first time is 2 to 5 hours.

[0017] In some embodiments, the second time is greater than or equal to 20 hours.

[0018] In some embodiments, the second time is 20 hours to 48 hours.

[0019] In some embodiments, the second time is 20-26 hours.

[0020] In some embodiments, the second time is 24-26 hours.

[0021] In some embodiments, after the heat preservation is completed, the mixture is cooled to 20°C-25°C.

[0022] In some embodiments, the preparation method further includes adjusting the pH of the mixture obtained after mixing the base liquid and the wetting liquid to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

[0023] In some embodiments, the pH of the mixed system is adjusted to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

[0024] In some embodiments, the preparation method of the base solution includes: heating water to 40°C-50°C, adding the suspending agent, stirring until dissolved, and then adding the buffer salt to prepare the base solution.

[0025] In some embodiments, the base liquid further comprises a sweetener, and the preparation method of the base liquid further comprises: adding the sweetener after the buffer salt has dissolved and continuing to stir until the sweetener dissolves.

[0026] In some embodiments, the preparation method of the base solution further includes: measuring the pH value of the base solution and adjusting the pH value of the base solution to 6.5-7.2.

[0027] In some embodiments, the pH of the base solution is adjusted to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

[0028] In some embodiments, the method for preparing the wetting solution includes: mixing glycerin and water, adding the nystatin and stirring until the nystatin is wetted, and then homogenizing until no particles are visible to the naked eye, thereby preparing the wetting solution.

[0029] In some embodiments, the mass ratio of glycerol to water is 1:(0.5-2).

[0030] In some embodiments, the preparation method further includes: pre-preparing the preservative into a preservative solution.

[0031] In some embodiments, the method for preparing the preservative solution includes: adding the preservative to an ethanol solution and stirring until the preservative dissolves.

[0032] In some embodiments, the preparation method further includes: adjusting the pH of the mixed system to 6.5-7.2, and then using water to bring the volume to the target volume.

[0033] In some embodiments, the composition comprising nystatin includes an oral suspension.

[0034] In some embodiments, the suspending agent comprises sodium carboxymethyl cellulose, the buffer salt comprises anhydrous disodium hydrogen phosphate, the preservative comprises methylparaben and propylparaben, and the flavoring agent comprises peppermint flavoring and cherry flavoring.

[0035] The aforementioned method for preparing the nystatin-containing composition employs a temperature control measure, first maintaining the temperature at a low temperature of 10℃-15℃ and then at 45℃-50℃. This significantly accelerates the crystallization and crystal growth rate of nystatin, thereby improving the stability and preparation efficiency of the composition and reducing production costs. Furthermore, the method's sequential addition of preservatives and flavoring agents after the temperature control period helps avoid instability reduction in preservatives due to the high-temperature holding process and volatilization of flavoring agents, further enhancing the stability of the nystatin-containing composition. Attached Figure Description

[0036] To better describe and illustrate the embodiments or examples provided in this application, reference may be made to one or more accompanying drawings. Additional details or examples used to describe the drawings should not be considered as limiting the scope of any of the disclosed applications, the currently described embodiments or examples, or the best mode of conduct of these applications as currently understood. Furthermore, the same reference numerals denote the same parts throughout the drawings. In the drawings:

[0037] Figure 1 The results of microscopic observation of nystatin raw materials.

[0038] Figure 2 The X-ray diffraction pattern of nystatin raw material.

[0039] Figure 3 The results are from microscopic observation of the reference formulation.

[0040] Figure 4 The X-ray diffraction pattern is for the reference formulation.

[0041] Figure 5 The results are from microscopic observations of Comparative Example 9 after 0 days of setting out.

[0042] Figure 6 The results are shown in the microscope after 30 days of observation for Comparative Example 9.

[0043] Figure 7 The X-ray diffraction patterns of Comparative Example 9 at 0, 4, 6, 9 and 12 days of setup are shown. Detailed Implementation

[0044] To facilitate understanding of the present invention, a more complete description will be given below with reference to relevant embodiments. Preferred embodiments of the present invention are shown below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that a thorough and complete understanding of the disclosure of the present invention will be achieved.

[0045] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0046] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0047] In this invention, numerical ranges are involved. Unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe features or characteristics, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0048] This document only specifically discloses some numerical ranges. However, any lower limit can be combined with any upper limit to form an unspecified range; and any lower limit can be combined with other lower limits to form an unspecified range, just as any upper limit can be combined with any other upper limit to form an unspecified range. Furthermore, each individually disclosed point or single value can itself serve as a lower or upper limit and be combined with any other point or single value or with other lower or upper limits to form an unspecified range.

[0049] Unless otherwise specified, the temperature parameters in this invention can be either constant temperature processing or processing within a certain temperature range. The constant temperature processing allows temperature fluctuations within the precision range controlled by the instrument.

[0050] In the description of the invention, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0051] Unless otherwise specified, all embodiments and optional embodiments of the present invention can be combined with each other to form new technical solutions. Unless otherwise specified, all technical features and optional technical features of the present invention can be combined with each other to form new technical solutions.

[0052] Unless otherwise specified, all steps of the present invention may be performed sequentially or randomly, but are preferably performed sequentially.

[0053] Oral administration is considered the most important and convenient method for maximizing the effect of drug molecules. Liquid dosage forms are the most common and widely accepted dosage forms, offering faster absorption than solid dosage forms, a better taste, making them a better choice for children and elderly patients, greater flexibility in achieving appropriate dosages, and convenience for patients who have difficulty swallowing other oral dosage forms.

[0054] Nystatin film-coated tablets (500,000 IU / tablet) are the only oral administration product in China to have passed the consistency evaluation for dosage form and specifications. However, when administering nystatin film-coated tablets to infants and young children, the dosage is difficult to control accurately, and the dispensing process damages the film coating, resulting in a strong earthy taste that can cause significant problems for administration. Furthermore, many patients, especially children and the elderly, find it more difficult to swallow tablets than solutions. However, nystatin oral suspensions offer certain advantages over tablets, allowing for accurate dosage control when administered in divided doses. However, traditional oral suspension preparation processes suffer from long production times and low efficiency; therefore, there is an urgent need for a method that provides a highly efficient and stable oral suspension preparation method.

[0055] Unstable crystal form or crystal morphology of a pharmaceutical formulation can significantly alter key properties such as solubility, dissolution rate, stability, and bioavailability, thereby affecting efficacy and safety. Therefore, it is essential to ensure that the crystal form and even the crystal morphology of the pharmaceutical formulation are consistent with the original formulation (trade name: Mycostatin). ® The product (manufacturer: Substipharm) maintains consistency and stability. Nystatin has three crystal forms: A, B, and C. Crystal form A and crystal form B (hydrated crystal form) can transform into each other under varying environmental humidity. Currently, the main crystal form of nystatin is crystal form A. During the preparation of nystatin oral suspension, nystatin transforms from crystal form A to hydrated crystal form B upon contact with water. Based on the above issues, this application regulates the crystal transformation and crystal growth rate of nystatin by controlling the solution temperature. Specifically, by controlling the solution temperature during preparation, nystatin achieves a state with the same crystal form and similar particle size distribution as the original formulation, thereby ensuring that the prepared nystatin oral suspension has similar dissolution rates to the reference formulation in a pH 4.5 dissolution medium.

[0056] One or more embodiments of this application provide a method for preparing a composition containing nystatin, comprising the following steps: preparing a base solution containing a suspending agent and a buffer salt; preparing a wetting solution containing nystatin; cooling the base solution to 10°C-15°C and then adding the wetting solution to obtain a mixture; keeping the mixture at 10°C-15°C for a first time, and then at 45°C-50°C for a second time; after the heat preservation is completed, cooling the mixture, then adding a preservative and a flavoring agent, and adjusting the pH value of the mixed system to 6.5-7.2 to prepare a composition containing nystatin.

[0057] First, cool the base liquid to 10℃-15℃ and then add the wetting solution. Since the formation of hydrates is exothermic, the base liquid should be cooled to the above range first. Nystatin in the wetting solution can quickly begin to form hydrates in a low-temperature environment.

[0058] Holding the mixture at a low temperature of 10℃-15℃ accelerates the conversion of nystatin from crystal form A to hydrate crystal form B. Continuing to hold the mixture at a high temperature of 45℃-50℃ accelerates the growth of hydrate crystal form B. As an example, the holding temperature for the low-temperature holding of the mixture can be, but is not limited to, 10℃, 11℃, 12℃, 13℃, 14℃, 15℃, or any range between two of the above temperatures. The holding temperature for the high-temperature holding of the mixture can be, but is not limited to, 45℃, 46℃, 47℃, 48℃, 49℃, 50℃, or any range between two of the above temperatures.

[0059] Adjusting the pH of the mixed system to 6.5-7.2 is beneficial to improving the stability of the prepared composition containing nystatin. As an example, the pH of the mixed system can be 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, or any range between two of the above pH values, etc., without specific limitation.

[0060] Understandably, the preparation method of the nystatin-containing composition of this application, by employing a temperature control measure of first maintaining a low temperature of 10℃-15℃ and then maintaining a temperature of 45℃-50℃, significantly accelerates the crystallization and crystal growth rate of nystatin, thereby improving the stability and preparation efficiency of the composition and reducing production costs. Simultaneously, the method of adding preservatives and flavoring agents after the temperature control is completed helps to avoid the decrease in stability of preservatives due to the high-temperature maintenance process and the volatilization of flavoring agents due to the high-temperature maintenance process, thereby further improving the stability of the nystatin-containing composition.

[0061] In some embodiments, the first time is greater than or equal to 2 hours; thereby, it is beneficial for nystatin to convert from crystal form A to hydrate crystal form B.

[0062] In some alternative implementations, the initial time is 2 to 5 hours; for example, it can be, but is not limited to, 2 hours, 3 hours, 4 hours, 5 hours, or any range between two of the above times. This facilitates the conversion of nystatin from crystal form A to hydrate crystal form B, while simultaneously improving production efficiency.

[0063] As one possible implementation, the second time is greater than or equal to 20 hours; thus, it is beneficial for the growth of hydrate crystal type B crystal frenulum.

[0064] In some optional embodiments, the second time is 20-48 hours; for example, it can be, but is not limited to, 20 hours, 22 hours, 24 hours, 26 hours, 28 hours, 30 hours, 32 hours, 34 hours, 36 hours, 38 hours, 40 hours, 42 hours, 44 hours, 46 hours, 48 ​​hours, or any range between two of the above times. This improves production efficiency while facilitating the growth of hydrate crystal form B crystal frenulum. Optionally, the second time is 20-26 hours. More optionally, the second time is 24-26 hours.

[0065] As one possible implementation, after the heat preservation is completed, the mixture is cooled to 20℃-25℃; for example, but not limited to 20℃, 21℃, 22℃, 23℃, 24℃, 25℃, or any two of the above temperatures. This helps to improve the stability of subsequently added preservatives and reduce the volatilization of flavoring agents.

[0066] In some embodiments, the preparation method further includes adjusting the pH of the mixture obtained after mixing the base solution and the wetting solution to 6.5-7.2 using hydrochloric acid or sodium hydroxide. This helps ensure the stability of nystatin and subsequently added preservatives within this pH range.

[0067] As one possible implementation method, the pH of the mixed system is adjusted to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

[0068] In some embodiments, the preparation method of the base solution includes: heating water to 40°C-50°C, adding a suspending agent, stirring until dissolved, and then adding a buffer salt to prepare the base solution.

[0069] As an example, water can be heated to 40°C, 42°C, 44°C, 46°C, 48°C, 50°C, or any range between two of the above temperatures.

[0070] Optionally, the stirring speed is 150 rpm to 350 rpm; for example, it can be, but is not limited to, 150 rpm, 200 rpm, 250 rpm, 300 rpm, 350 rpm, or any range between two of the above speeds.

[0071] In some embodiments, the base liquid also contains a sweetener, and the preparation method of the base liquid further includes: adding the sweetener after the buffer salt has dissolved and continuing to stir until the sweetener dissolves. Optionally, the sweetener includes sucrose.

[0072] In some optional embodiments, the preparation method of the base solution further includes: measuring the pH value of the base solution and adjusting the pH value of the base solution to 6.5-7.2; for example, it can be, but is not limited to, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, or any range between two of the above pH values. Thus, a pH environment favorable to nystatin is maintained before the wetting solution is added.

[0073] Optionally, the pH of the base solution can be adjusted to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

[0074] In some embodiments, the method for preparing the wetting solution includes: mixing glycerin and water, adding nystatin and stirring until the nystatin wets the solution, and then homogenizing it until no particles are visible to the naked eye, thereby preparing the wetting solution.

[0075] As a non-limiting example, the homogenization speed is 5000rpm-15000rpm; for example, it can be, but is not limited to, 5000rpm, 6000rpm, 8000rpm, 10000rpm, 12000rpm, 14000rpm, 15000rpm, or any range between two of the above speeds.

[0076] In some alternative embodiments, the mass ratio of glycerol to water is 1:(0.5-2); for example, it can be, but is not limited to, 1:0.5, 1:0.6, 1:0.8, 1:1, 1:1.2, 1:1.4, 1:1.6, 1:1.8, 1:2, or any range between any two of the above mass ratios.

[0077] In some embodiments, the method for preparing a composition containing nystatin further includes: pre-formulating the preservative into a preservative solution.

[0078] In some alternative embodiments, the method for preparing the preservative solution includes: adding the preservative to an ethanol solution and stirring until the preservative dissolves.

[0079] In some embodiments, the preparation method of the composition containing nystatin further includes: adjusting the pH of the mixed system to 6.5-7.2 and then making up to the target volume with water.

[0080] In some of these embodiments, the suspending agent includes sodium carboxymethyl cellulose.

[0081] As one possible implementation, the buffer salt includes anhydrous disodium hydrogen phosphate.

[0082] In some alternative implementations, the preservatives include methylparaben and propylparaben.

[0083] In some exemplary embodiments, flavoring agents include peppermint flavoring and cherry flavoring.

[0084] In some embodiments, the composition containing nystatin includes an oral suspension.

[0085] In some embodiments, the raw materials for preparing the composition containing nystatin include: nystatin, solvent, sweetener, preservative, suspending agent, pH adjuster, buffer salt, and flavoring agent; the solvent includes anhydrous ethanol, glycerol, and water; the sweetener includes sucrose; the preservative includes methylparaben and propylparaben; the suspending agent includes sodium carboxymethyl cellulose; the pH adjuster includes hydrochloric acid and sodium hydroxide; the buffer salt includes anhydrous disodium hydrogen phosphate; and the flavoring agent includes peppermint flavor and cherry flavor. A method for preparing the composition containing nystatin includes:

[0086] Step 1, Preparation of base solution: Take 40% of the volume of purified water, stir and heat to 40℃-50℃, then add sodium carboxymethyl cellulose and continue stirring until it is completely dissolved (visually no white particles or transparent gel-like clumps are visible); add anhydrous disodium hydrogen phosphate and continue stirring to dissolve; add sucrose and stir until completely dissolved, measure the pH value and adjust the pH value to 6.5-7.2 using 1.0 mol / L hydrochloric acid solution or NaOH solution.

[0087] Step 2, wetting solution preparation: Mix glycerol and purified water at a weight ratio of 1:1, add nystatin and stir until nystatin is wetted (no solid-liquid separation is visible on the liquid surface). Then turn on the high-shear homogenizer to homogenize until no particles are visible, so that nystatin is fully wetted and dispersed.

[0088] Step 3, Preparation of preservative solution: Add methylparaben and propylparaben to ethanol and stir until the preservative is completely dissolved.

[0089] Step 4, Mixing and Heating: Continue stirring the base solution and cool it to 10℃-15℃. Then slowly add the wetting solution and stir for more than 10 minutes. Then adjust the pH value to 6.5-7.2 with 1.0mol / L hydrochloric acid solution or NaOH solution. Continue stirring at 10℃-15℃ for more than 2 hours. Control the same stirring speed and heat to 45℃-50℃ and maintain for more than 20 hours. Then cool it to room temperature (20℃-25℃), add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for more than 10 minutes.

[0090] Step 5, Adjust the volume: Adjust the pH to 6.5-7.2 with 1.0 mol / L hydrochloric acid solution or NaOH solution, and then adjust the volume to the target preparation volume with purified water.

[0091] Step 6, Filling: Fill the above-mentioned solution to 60ml HDPE bottles and add child-safe caps.

[0092] Compositions containing nystatin, by weight and volume percentage, include:

[0093] Nystatin 0.8%-3%,

[0094] Anhydrous ethanol 0.5%-1.5%,

[0095] Sucrose 30%-65%,

[0096] Methylparaben 0.05%-0.2%,

[0097] Propylparaben 0.01%-0.1%

[0098] Sodium carboxymethyl cellulose 0.01%-0.2%,

[0099] Glycerin 5%-20%,

[0100] Anhydrous disodium hydrogen phosphate 0.1%-0.5%,

[0101] Peppermint flavoring 0.01%-0.5%,

[0102] Cherry flavoring 0.01%-0.5%,

[0103] Appropriate amount of hydrochloric acid

[0104] Sodium hydroxide in appropriate amount, and

[0105] The remainder is water.

[0106] Optionally, the composition comprising nystatin, by weight / volume percentage, includes:

[0107] Nystatin 1.64%,

[0108] Anhydrous ethanol 0.76%,

[0109] 50% sucrose

[0110] Methylparaben 0.1%,

[0111] Propylparaben 0.02%,

[0112] Sodium carboxymethyl cellulose 0.047%,

[0113] 10% glycerin

[0114] Anhydrous disodium hydrogen phosphate 0.29%,

[0115] Peppermint flavoring 0.05%,

[0116] Cherry flavoring 0.05%,

[0117] Appropriate amount of hydrochloric acid

[0118] Sodium hydroxide in appropriate amount, and

[0119] The remainder is water.

[0120] It should be noted that the mass-volume percentage of each component is calculated based on the mass m (in g) of each component and the volume V (in mL) of the composition containing nystatin, using the formula m / V×100%, the same below.

[0121] The technical solutions of this application will be described in detail below with reference to specific embodiments. It should be understood that these embodiments are only for illustrating this application and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application first, or follow experimental manuals or conventional conditions in the field, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the field.

[0122] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0123] Unless otherwise specified, all raw materials used in the following examples and comparative examples are commercially available or prepared using conventional methods. The sources of the raw materials used in the following examples and comparative examples are shown in Table 1.

[0124] Table 1

[0125]

[0126] Example 1

[0127] 1. The prescription composition is shown in Table 2.

[0128] Table 2

[0129]

[0130] 2. Preparation process

[0131] Step 1, Preparation of base solution: Take 40% of the volume of purified water, stir and heat to 45℃, then add sodium carboxymethyl cellulose and continue stirring until it is completely dissolved (visually no white particles or transparent gel-like clumps are visible); add anhydrous disodium hydrogen phosphate and continue stirring to dissolve; add sucrose and stir until completely dissolved, measure the pH value and adjust the pH value to 7.03 using 1.0 mol / L hydrochloric acid solution.

[0132] Step 2, wetting solution preparation: Mix glycerol and purified water at a weight ratio of 1:1, add nystatin and stir until nystatin is wetted (no solid-liquid separation is visible on the liquid surface). Then turn on the high-shear homogenizer to homogenize until no particles are visible, so that nystatin is fully wetted and dispersed.

[0133] Step 3, Preparation of preservative solution: Add methylparaben and propylparaben to ethanol and stir until the preservative is completely dissolved.

[0134] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 15°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH value to 7.00 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 15°C for 2 hours. Control the same stirring speed and heat to 50°C for 24 hours. Then cool it to room temperature of 25°C, add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0135] Step 5, Adjust the volume: Adjust the pH to 7.00 with 1.0 mol / L NaOH solution, and then adjust the volume to the target preparation volume with purified water.

[0136] Step 6, Filling: Fill the above-mentioned solution after adjusting the volume into 60ml HDPE bottles and add child safety caps to obtain oral suspension.

[0137] Example 2

[0138] 1. The prescription composition of Example 2 is the same as that of Example 1.

[0139] 2. Preparation process

[0140] Steps 1-3 and 5-6 in Example 2 are the same as steps 1-3 and 5-6 in Example 1, except that the high-temperature insulation temperature in step 4 of Example 2 is 45℃; all other aspects are the same. Step 4 of Example 2 is detailed below:

[0141] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 15°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH value to 6.99 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 15°C for 2 hours. Control the same stirring speed and heat to 45°C for 24 hours. Then cool it to room temperature of 25°C, add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0142] Example 3

[0143] 1. The prescription composition in Example 3 is the same as that in Example 1.

[0144] 2. Preparation process

[0145] Steps 1-3 and 5-6 in Example 3 are the same as steps 1-3 and 5-6 in Example 2, except that the low-temperature insulation temperature in step 4 of Example 3 is 10℃; all other aspects are the same. Step 4 of Example 3 is detailed below:

[0146] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 10°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH to 7.01 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 10°C for 2 hours. Maintain the same stirring speed and heat to 45°C for 24 hours. Then cool it to room temperature (25°C), add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0147] Example 4

[0148] 1. The prescription composition in Example 4 is the same as that in Example 1.

[0149] 2. Preparation process

[0150] Steps 1-3 and 5-6 in Example 4 are the same as steps 1-3 and 5-6 in Example 3, except that the low-temperature holding time in step 4 of Example 4 is 5 hours; all other steps are the same. Step 4 of Example 4 is detailed below:

[0151] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 10°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH to 7.01 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 10°C for 5 hours. Maintain the same stirring speed and heat to 45°C for 24 hours. Then cool it to room temperature (25°C), add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0152] Comparative Example 1

[0153] 1. The prescription composition of Comparative Example 1 is the same as that of Example 1.

[0154] 2. Preparation process

[0155] Steps 1-3 and 5-6 of Comparative Example 1 are the same as steps 1-3 and 5-6 of Example 1, except that: in Comparative Example 1, low-temperature insulation was not performed in step 4, and the high-temperature insulation temperature was 40°C. Step 4 of Comparative Example 1 is as follows:

[0156] Step 4, Mixing and Keeping Warm: Continue stirring the base liquid, then slowly add the wetting liquid and stir for 10 minutes. Then adjust the pH to 7.00 with 1.0 mol / L hydrochloric acid solution. Maintain the same stirring speed and heat to 40°C for 24 hours. Then cool down to room temperature (25°C), add preservative, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0157] Comparative Example 2

[0158] 1. The prescription composition of Comparative Example 2 is the same as that of Example 1.

[0159] 2. Preparation process

[0160] Steps 1-3 and 5-6 of Comparative Example 2 are the same as steps 1-3 and 5-6 of Example 1, except that: in Comparative Example 2, low-temperature insulation was not performed in step 4, and the high-temperature insulation temperature was 45°C. Step 4 of Comparative Example 2 is as follows:

[0161] Step 4, Mixing and Keeping Warm: Continue stirring the base liquid, then slowly add the wetting liquid and stir for 10 minutes. Then adjust the pH to 7.00 with 1.0 mol / L hydrochloric acid solution. Maintain the same stirring speed and heat to 45°C for 24 hours. Then cool down to room temperature (25°C), add preservative, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0162] Comparative Example 3

[0163] 1. The prescription composition of Comparative Example 3 is the same as that of Example 1.

[0164] 2. Preparation process

[0165] Steps 1-3 and 5-6 of Comparative Example 3 are the same as steps 1-3 and 5-6 of Example 1, except that low-temperature insulation was not performed in step 4 of Comparative Example 3. Step 4 of Comparative Example 3 is as follows:

[0166] Step 4, Mixing and Keeping Warm: Continue stirring the base solution, then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH to 7.00 with 1.0 mol / L hydrochloric acid solution. Maintain the same stirring speed and heat to 50°C for 24 hours. Then cool down to room temperature (25°C), add preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0167] Comparative Example 4

[0168] 1. The prescription composition of Comparative Example 4 is the same as that of Example 1.

[0169] 2. Preparation process

[0170] Steps 1-3 and 5-6 of Comparative Example 4 are the same as steps 1-3 and 5-6 of Example 1, except that the low-temperature insulation temperature in step 4 of Comparative Example 4 is 20°C. Step 4 of Comparative Example 4 is detailed below:

[0171] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 20°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH value to 7.01 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 20°C for 2 hours. Control the same stirring speed and heat to 50°C for 24 hours. Then cool it to room temperature of 25°C, add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0172] Comparative Example 5

[0173] 1. The prescription composition of Comparative Example 5 is the same as that of Example 1.

[0174] 2. Preparation process

[0175] Steps 1-3 and 5-6 of Comparative Example 5 are the same as steps 1-3 and 5-6 of Example 1, except that the low-temperature insulation temperature in step 4 of Comparative Example 5 is 7°C. Step 4 of Comparative Example 5 is as follows:

[0176] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 7°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH value to 7.00 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 7°C for 2 hours. Control the same stirring speed and heat to 50°C for 24 hours. Then cool it to room temperature of 25°C, add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0177] It should be noted that, due to the high solid and glycerol content in this comparative example, the viscosity is too high below 10°C, making stirring difficult. Furthermore, excessively low temperature control is detrimental to preparation efficiency and production cost control.

[0178] Comparative Example 6

[0179] 1. The prescription composition of Comparative Example 6 is the same as that of Example 1.

[0180] 2. Preparation process

[0181] Steps 1-3 and 5-6 of Comparative Example 6 are the same as steps 1-3 and 5-6 of Example 1, except that the high-temperature holding temperature in step 4 of Comparative Example 6 is 55°C. Step 4 of Comparative Example 6 is as follows:

[0182] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 15°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH value to 7.03 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 15°C for 2 hours. Control the same stirring speed and heat to 55°C for 24 hours. Then cool it to room temperature of 25°C, add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0183] It should be noted that excessively high temperatures can cause nystatin to become unstable. After maintaining the temperature at 55°C for 24 hours, the RRT1.27 impurity in the formulation increased from 0.37 in the active pharmaceutical ingredient to 1.19, which may affect the safety and efficacy of the product.

[0184] Comparative Example 7

[0185] 1. The prescription composition of Comparative Example 7 is the same as that of Example 1.

[0186] 2. Preparation process

[0187] Steps 1-3 and 5-6 of Comparative Example 7 are the same as steps 1-3 and 5-6 of Example 1, except that the high-temperature holding temperature in step 4 of Comparative Example 7 is 40°C. Step 4 of Comparative Example 7 is as follows:

[0188] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 15°C. Then slowly add the wetting solution and stir for 10 minutes. Then adjust the pH value to 7.00 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 15°C for 2 hours. Control the same stirring speed and heat to 40°C for 24 hours. Then cool it to room temperature of 25°C, add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 10 minutes.

[0189] The oral suspensions prepared in Examples 1-4, Comparative Examples 1-4, and Comparative Example 7, as well as the reference formulation (nystatin oral suspension, trade name: Mycostatin) ® Dissolution testing was performed on a sample (manufacturer: Substipharm, batch number 0470924). The dissolution testing method is as follows:

[0190] The dissolution conditions were as follows: 900 ml of a buffer solution of sodium acetate trihydrate and glacial acetic acid at pH 4.5 was used as the dissolution medium; the paddle method was controlled at a rotation speed of 75 rpm; and the sampling times were 5 min, 10 min, 15 min, 30 min, 45 min, and 60 min.

[0191] It should be noted that the criterion for judging whether dissolution is similar is as follows: According to the "Guidelines for Bioequivalence Testing of Generic Drugs (Japan) - 2020.03.19", when the average dissolution rate of the reference formulation does not exceed 10% within the specified dissolution time, the average dissolution rate of the test formulation within the specified testing time only needs to be within ±9% (absolute value) of the average dissolution rate of the reference formulation. In this case, the dissolution of the test formulation and the reference formulation is considered similar.

[0192] The differences in preparation and dissolution results of Examples 1-4, Comparative Examples 1-4 and Comparative Example 7 are shown in Table 3.

[0193] Table 3

[0194]

[0195] As can be seen from the comparison of the results of Comparative Examples 1-3 in Table 3, the higher the temperature of the high-temperature insulation, the lower the dissolution rate. That is, the higher the temperature of the high-temperature insulation, the more similar the dissolution rate is to the reference preparation.

[0196] As can be seen from the comparison of the results of Examples 1-2 and Comparative Example 4 in Table 3, the temperature of the low-temperature insulation has a more significant effect on the reduction of dissolution.

[0197] As can be seen from the comparison of the results of Examples 3-4 in Table 3, when the low temperature is 10℃ and the holding time is greater than or equal to 2 hours, extending the holding time has no significant effect on the reduction of dissolution.

[0198] A comparison of the results from Example 1 and Comparative Example 4 shows that when the temperature is kept low within the range of 10℃-15℃, the decrease in dissolution is more significant at lower temperatures. If the temperature is kept low below 10℃, stirring will be difficult due to excessive viscosity. From the perspective of operability in the production workshop, it is more difficult to achieve the sample temperature (below 10℃) and the production cost will be higher (longer cooling time and higher equipment requirements).

[0199] A comparison of the results from Examples 1-2 and Comparative Example 7 shows that when performing high-temperature insulation within the temperature range of 45℃-50℃, the higher the temperature, the lower the dissolution rate. If high-temperature insulation is performed above 50℃, there is a potential risk to product stability due to the excessively high temperature (above 50℃), and higher temperatures also result in longer heating and cooling times. Therefore, the suitable temperature range for high-temperature insulation is 45℃-50℃.

[0200] Example 5

[0201] 1. The prescription composition in Example 5 is the same as that in Example 1.

[0202] 2. Preparation process

[0203] Step 1, Preparation of base solution: Take 40% of the volume of purified water, stir and heat to 42℃, then add sodium carboxymethyl cellulose and continue stirring until it is completely dissolved (visually no white particles or transparent gel-like clumps are visible); add anhydrous disodium hydrogen phosphate and continue stirring to dissolve; add sucrose and stir until completely dissolved, measure the pH value and adjust the pH value to 7.01 with 1.0 mol / L hydrochloric acid solution.

[0204] Step 2, wetting solution preparation: Mix glycerol and purified water at a weight ratio of 1:1, add nystatin and stir until nystatin is wetted (no solid-liquid separation is visible on the liquid surface). Then turn on the high-shear homogenizer to homogenize until no particles are visible, so that nystatin is fully wetted and dispersed.

[0205] Step 3, Preparation of preservative solution: Add methylparaben and propylparaben to ethanol and stir until the preservative is completely dissolved.

[0206] Step 4, Mixing and Keeping Warm: Continue stirring the base solution and cool it to 12.0℃. Then slowly add the wetting solution and stir for 12 minutes. Then adjust the pH value to 7.00 with 1.0 mol / L hydrochloric acid solution. Continue stirring at 12.0℃ for about 2.1 hours. Control the same stirring speed and heat to 46.0℃ for 20 hours. Then cool it to room temperature, add the preservative solution, peppermint flavoring, and cherry flavoring, and continue stirring for 21 minutes.

[0207] Step 5, Adjust the volume: Adjust the pH to 7.00 with 1.0 mol / L NaOH solution, and then adjust the volume to the target preparation volume with purified water.

[0208] Step 6, Filling: Fill the above-mentioned solution to 60ml HDPE bottles and add child-safe caps.

[0209] Example 6

[0210] 1. The prescription composition of Example 6 is the same as that of Example 1.

[0211] 2. Preparation process

[0212] Steps 1-3 and 5-6 in Example 6 are the same as steps 1-3 and 5-6 in Example 5, except that the high-temperature insulation time in step 4 of Example 6 is 22 hours; all other steps are the same.

[0213] Example 7

[0214] 1. The prescription composition in Example 7 is the same as that in Example 1.

[0215] 2. Preparation process

[0216] Steps 1-3 and 5-6 in Example 7 are the same as steps 1-3 and 5-6 in Example 5, except that the high-temperature insulation time in step 4 of Example 7 is 24 hours; all other steps are the same.

[0217] Example 8

[0218] 1. The prescription composition of Example 8 is the same as that of Example 1.

[0219] 2. Preparation process

[0220] Steps 1-3 and 5-6 in Example 8 are the same as steps 1-3 and 5-6 in Example 5, except that the high-temperature insulation time in step 4 of Example 8 is 26 hours; all other steps are the same.

[0221] Comparative Example 8

[0222] 1. The prescription composition of Comparative Example 8 is the same as that of Example 1.

[0223] 2. Preparation process

[0224] Steps 1-3 and 5-6 of Comparative Example 8 are the same as steps 1-3 and 5-6 of Example 5, except that the high-temperature holding time in step 4 of Comparative Example 8 is 15 hours; all other steps are the same.

[0225] Dissolution tests were performed on the oral suspensions prepared in Examples 5-8 and Comparative Example 8. The dissolution test method was as described above. The differences in preparation of Examples 5-8 and Comparative Example 8 and the dissolution results are shown in Table 4.

[0226] Table 4

[0227]

[0228] As can be seen from the comparison of the results of Examples 5-8 and Comparative Example 8 in Table 4, after the high temperature holding time reaches 20 hours, extending the holding time has no significant effect on the reduction of dissolution.

[0229] Example 7 and the reference formulation (nystatin oral suspension, trade name: Mycostatin) ®The stability of the sample (manufacturer: Substipharm, batch number 0470924) was sampled and then dissolution was tested, using the method described above. The dissolution results of Example 7 and the reference formulation after sample release are shown in Table 5.

[0230] Table 5

[0231]

[0232] As shown in Table 5, the oral suspension prepared in this application has a similar dissolution rate to the reference formulation throughout the stability study; this indicates that the heat preservation process of this application not only improves production efficiency (eliminating the need for storage for several days) but also ensures the stability of product quality during the stability study.

[0233] The product stability study results for Example 7 and the reference formulation are shown in Table 6.

[0234] Table 6

[0235]

[0236] Comparative Example 9

[0237] 1. The prescription composition of Comparative Example 9 is the same as that of Example 1.

[0238] 2. Preparation process

[0239] Steps 1-3 and 5-6 of Comparative Example 9 are the same as steps 1-3 and 5-6 of Example 1, except that low-temperature insulation and high-temperature insulation were not performed in step 4 of Comparative Example 9. Step 4 of Comparative Example 9 is as follows:

[0240] Step 4: Mix well: Control the stirring speed of the base liquid at 250 rpm and cool it to room temperature. Then slowly add the wetting liquid and stir for 10 minutes. Then add the preservative liquid, peppermint flavoring and cherry flavoring and continue stirring for 10 minutes.

[0241] Stability testing was performed on Comparative Example 9, followed by particle size and dissolution testing. The dissolution testing method was as described above. The particle size analysis equipment used was from Newpatek, model HELOS / BR. Before testing, the sample was shaken for 30 seconds, with water as the dispersion medium, to ensure the optical concentration was between 15% and 25%. The sample was stirred at 1900 rpm and collected for 10 seconds.

[0242] At the same time, the reference preparation (nystatin oral suspension, trade name: Mycostatin) was also tested. ®The particle size and dissolution of the sample (manufacturer: Substipharm, batch number 0470924) were tested. The particle size and dissolution results of Comparative Example 9 after sample release, as well as the particle size and dissolution results of the reference formulation, are shown in Table 7. In Table 7, "room temperature" refers to a sample release temperature of 21℃-22℃.

[0243] Table 7

[0244]

[0245] As shown in Table 7, the oral suspension of Comparative Example 9 has similar dissolution rate to the reference preparation in a pH 4.5 dissolution medium when stored at room temperature for more than 20 days, and the dissolution rate is consistent with the reference preparation when stored for more than 30 days. However, for products that need to be commercially produced, the storage time is too long, which affects production efficiency and increases product production costs. Therefore, the process needs to be improved and optimized.

[0246] Electron microscopy and X-ray diffraction were used to observe the reference preparation (nystatin oral suspension, trade name: Mycostatin®, manufacturer: Substipharm, batch number 0470924), comparative example 9, and nystatin raw material, respectively. Comparative example 9 was observed under a microscope after 0 days and 30 days of storage at room temperature; X-ray diffraction analysis was performed on comparative example 9 after 0, 4, 6, 9, and 12 days of storage at room temperature.

[0247] The microscopic observation results and X-ray diffraction patterns of nystatin raw materials are as follows: Figure 1 and Figure 2 As shown. The microscopic observation results and X-ray diffraction patterns of the reference formulation are as follows. Figure 3 and Figure 4 As shown in the figure. The microscopic observation results of Comparative Example 9 after 0 days and 30 days of placement at room temperature are respectively shown in the figure. Figure 5 and Figure 6 As shown. The X-ray diffraction patterns of Comparative Example 9 at 0, 4, 6, 9, and 12 days of setup are as follows. Figure 7 As shown.

[0248] Depend on Figure 1 , Figure 3 and Figure 5-6 The comparison showed that the crystal structure of the active pharmaceutical ingredient (API) did not change after being prepared into a formulation sample. However, with prolonged storage, the crystal structure of the formulation sample grew from short rod-shaped or granular to needle-shaped, consistent with the reference formulation. Figure 2 , Figure 4 and Figure 7 The comparison shows that the crystal form of the active pharmaceutical ingredient (API) changed after being prepared into a formulation sample, and the changes were consistent with the reference formulation during storage. Figure 7It can be seen that Comparative Example 9 had the same crystal form during the layout process, and the crystal form did not change.

[0249] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0250] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A method of preparing a composition comprising nystatin, characterized in that, The method comprises the following steps: preparing a base solution containing a suspending agent and a buffer salt; preparing a wetting solution containing nystatin; cooling the base solution to 10-15°C and then adding the wetting solution to prepare a mixed solution; keeping the mixed solution at 10-15°C for a first time and then keeping it at 45-50°C for a second time; after the keeping, cooling the mixed solution, adding a preservative and a flavoring agent, and adjusting the pH of the mixed system to 6.5-7.2 to prepare the composition containing nystatin.

2. The method of preparing a composition comprising nystatin according to claim 1, wherein, The first time is greater than or equal to 2 hours. Optionally, the first time is 2-5 hours; and / or, The second time is greater than or equal to 20 hours. Optionally, the second time is 20-48 hours, optionally 20-26 hours, and further optionally 24-26 hours.

3. The method of preparing a composition comprising nystatin according to claim 1, wherein, After the keeping, the mixed solution is cooled to 20-25°C.

4. The method of preparing a composition comprising nystatin according to claim 1, wherein, The method further comprises adjusting the pH of the mixed solution obtained by mixing the base solution and the wetting solution to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

5. The method of preparing a composition comprising nystatin according to claim 1, wherein, The pH of the mixed system is adjusted to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

6. The method of preparing a composition comprising nystatin according to claim 1, wherein, The method for preparing the base solution comprises: heating water to 40-50°C, adding the suspending agent, stirring until it is dissolved, and then adding the buffer salt to prepare the base solution; Optionally, the base solution further contains a sweetener, and the method for preparing the base solution further comprises: after the buffer salt is dissolved, adding the sweetener and continuing to stir until the sweetener is dissolved. Optionally, the method for preparing the base solution further comprises: measuring the pH of the base solution and adjusting the pH of the base solution to 6.5-7.2; and further optionally, adjusting the pH of the base solution to 6.5-7.2 using hydrochloric acid or sodium hydroxide.

7. The method of preparing a composition comprising nystatin according to claim 1, wherein, The method for preparing the wetting solution comprises: mixing glycerol and water, adding nystatin and stirring until the nystatin is wetted, and then homogenizing until no particles are visually observed to prepare the wetting solution; Optionally, the mass ratio of the glycerol to the water is 1:(0.5-2).

8. The method of preparing a composition comprising nystatin according to claim 1, wherein, The method further comprises: preparing the preservative as a preservative solution in advance; Optionally, the method for preparing the preservative solution comprises: adding the preservative to an ethanol solution and stirring until the preservative is dissolved.

9. The method of preparing a composition comprising nystatin according to claim 1, wherein, The method further comprises: after adjusting the pH of the mixed system to 6.5-7.2, using water to make up to the target volume.

10. A method of preparing a composition comprising nystatin according to any one of claims 1 to 9, characterized in that, The suspending agent comprises sodium carboxymethyl cellulose, the buffer salt comprises anhydrous disodium hydrogen phosphate, the preservative comprises methylparaben and propylparaben, and the flavoring agent comprises peppermint flavor and cherry flavor; and / or, The composition containing nystatin comprises an oral suspension.

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