Process for preparing β-lactamase inhibitor liquid preparation

Abstract

A process for preparing a β-lactamase inhibitor liquid preparation. The preparation process comprises subjecting a suspension of a β-lactamase inhibitor to high-pressure homogenization to obtain the β-lactamase inhibitor liquid preparation, wherein the high-pressure homogenization is performed at a pressure of 1-30 Kpsi by using a high-pressure homogenizer or a micro-jet homogenizer as a device for 3-57 cycles. The β-lactamase inhibitor liquid preparation prepared by the preparation process is placed at 25℃ for 24 H under natural light conditions, and there is no significant change in the concentration and total impurity content.

Description

Process for the preparation of a beta-lactamase inhibitor liquid formulation TECHNICAL FIELD

[0001] The present invention relates to a process for the preparation of a beta-lactamase inhibitor liquid formulation. BACKGROUND

[0002] Beta-lactam antibiotics have been the first choice of antimicrobial agents in the clinic, but the efficacy of the related antibiotics is currently affected by bacterial beta-lactamase, which can produce resistance to penicillins, extended-spectrum cephalosporins, monobactams, and carbapenems. In order to solve the beta-lactamase-mediated resistance, beta-lactamase inhibitors were introduced into clinical practice, which greatly enhanced the efficacy of beta-lactam in the treatment of bacterial infections (Clin Microbiol Rev. 2010 Jan; 23(1): 160-201). Antibiotic resistance produced by beta-lactamase poses more challenges to the efficacy of beta-lactam drugs and the category of antibiotics used in the clinic. In the past three decades, only a few beta-lactamase inhibitors have been introduced into the market (Medchemcomm. 2018 Aug 17; 9(9): 1439-1456). Beta-lactamase inhibitors in the clinic are single-dose and multi-dose injectable preparations that can also be used in combination with other antibacterial drugs. However, the beta-lactamase inhibitor of the following general formula is a poorly soluble small molecule drug, and the drug is prone to precipitation during the storage and use of the injection, and has poor stability, which is not conducive to the combination with other antibacterial drugs.

[0003] The dissolution strategy of poorly soluble drugs is an important issue that needs to be considered in the application of the pharmaceutical industry. Researchers and professionals have explored many methods to try to solve such problems. For example, the dissolution of nateglinide (a lipophilic insoluble drug) is improved by nanoparticle and blending technology (Int J Pharm. 2013 Sep 15; 454(1): 562-7); the solubility of insoluble drugs is enhanced by liquid freezing into spray-micronized products to improve physical stability (Pharm Dev Technol. 2003; 8(2): 187-97), etc. Although these methods have been proven to increase the solubility of drugs, there are still great limitations in these methods today: only suitable for small-dose production, complex process method, long development period of formulation, etc. At present, relatively mature preparation processes need to be considered to better solve such problems.

[0004] High pressure homogenization process is mainly applied in biological and pharmaceutical industries. It is a process of breaking solid materials in nanoemulsion, liposome and nanosuspension by homogenizer to achieve superfine solid particles and form uniform suspension emulsion. It is achieved by mechanical forces such as high-speed shearing, high-frequency oscillation, cavitation phenomenon and convection impact, and corresponding thermal effect to improve the solubility of poorly soluble compounds and make the material in a good uniform distribution state.

[0005] The force of the homogenizer commonly used in the homogenization process is mainly shear force and pressure. The homogenizers commonly used in pharmaceutical and biological industries mainly include high-pressure homogenizer and microfluidic homogenizer. The high-pressure homogenizer mainly causes cavitation effect and turbulent effect by high pressure generated by the pressure system to extrude, stretch, impact and break the material. The disadvantages of the high-pressure homogenizer are: the design gap of the homogenizing valve is large, the homogenizing pressure is low, and it is easy to be damaged when homogenizing high-hardness particles, and the maintenance is difficult. The advantages are: the price is relatively low. The high-pressure homogenizer is suitable for processing soft and semi-soft granular materials. The microfluidic homogenizer uses a hole of about one hundred microns to form a supersonic jet, and the jets collide with each other to produce extremely strong shearing. The advantage is that it can produce better particle size distribution effect. The disadvantage is that the flow is small and the cost is relatively high. The high-pressure homogenizing cavity as the core component has a special internal geometry, which also affects the homogenization effect of the final product.

[0006] In recent years, nano-suspension as a kind of injection has attracted widespread attention. Improving the dissolution rate of nano-suspension can solve the problem of poor drug solubility. Nano-suspension has a large surface area, which not only increases the dissolution rate, but also increases the saturation of the solution, thereby improving the bioavailability. Therefore, commercial nano-suspension products such as Rapamune, Emend, etc. have been rapidly adopted by the pharmaceutical industry.(Pharmaceutical development and technology, 24(10), 1278-1286).

[0007] In view of the high impurity content and poor drug solubility of the liquid preparation prepared by the preparation method shown in FIG. 1, it is urgent to develop a new liquid preparation preparation process for β-lactamase inhibitor to improve the solubility and reduce the impurity content and stabilize the storage. Considering that the high-pressure homogenization technology has been mature, and the preparation development method of β-lactamase inhibitor using this technology has not been applied, the high-pressure homogenization process can be used to prepare β-lactamase inhibitor type broad-spectrum antibacterial poorly soluble small molecule compounds. SUMMARY

[0008] In order to overcome the high impurity content and poor drug solubility of the liquid preparation of the existing β-lactamase inhibitor, a preparation process of the β-lactamase inhibitor liquid preparation is provided. The β-lactamase inhibitor prepared by the preparation process of the present application has no obvious change in concentration and total impurity content after being placed at 25°C under natural light for 24H.

[0009] The present application provides a preparation process of a β-lactamase inhibitor liquid preparation, which comprises preparing a β-lactamase inhibitor liquid preparation from a suspension of a β-lactamase inhibitor by high-pressure homogenization,

[0010] The pressure of the high-pressure homogenization is 1-30Kpsi.

[0011] The high-pressure homogenization cycle is 3-57 times.

[0012] The equipment for high-pressure homogenization is a high-pressure homogenizer or a microfluidic homogenizer.

[0013] In an embodiment, the flow rate of the high-pressure homogenization is 50-200ml / min, such as 50ml / min, 100ml / min, 150ml / min or 200ml / min.

[0014] In an embodiment, the pressure of the high-pressure homogenization can be 5-25Kpsi, such as 5Kpsi, 10Kpsi, 15Kpsi, 20Kpsi or 25Kpsi.

[0015] In an embodiment, the high-pressure homogenization cycle is 3 times, 9 times, 18 times, 24 times, 27 times, 30 times, 35 times, 40 times, 45 times, 50 times or 57 times.

[0016] In an embodiment, when the pressure of the high-pressure homogenization is 5Kpsi, the high-pressure homogenization cycle is 3 times.

[0017] In an embodiment, when the pressure of the high-pressure homogenization is 10Kpsi, the high-pressure homogenization cycle is 3 times.

[0018] In an embodiment, when the pressure of the high-pressure homogenization is 20Kpsi, the high-pressure homogenization cycle is 3 times or 18 times.

[0019] In an embodiment, when the pressure of the high-pressure homogenization is 20Kpsi, the high-pressure homogenization cycle is 24 or 35 times.

[0020] In an embodiment, when the pressure of the high-pressure homogenization is 25Kpsi, the high-pressure homogenization cycle is 27, 30 or 57 times.

[0021] In one embodiment, the high pressure homogenization cycle is 3 cycles at 5 Kpsi, 3 cycles at 10 Kpsi, and 24 cycles at 20 Kpsi.

[0022] In one embodiment, the high pressure homogenization cycle is 3 cycles at 5 Kpsi, 3 cycles at 20 Kpsi, and 27 cycles at 25 Kpsi.

[0023] In one embodiment, the high pressure homogenization cycle is 3 cycles at 5 Kpsi, 3 cycles at 10 Kpsi, and 24 cycles at 20 Kpsi.

[0024] In one embodiment, the high pressure homogenization cycle is 3 cycles at 5 Kpsi, 3 cycles at 10 Kpsi, and 18 cycles at 20 Kpsi.

[0025] In the present application, the time of high pressure homogenization is the conventional homogenization time in the art. Preferably, the time of high pressure homogenization is related to the pressure and the number of cycles of high pressure homogenization. The higher the pressure of high pressure homogenization, the more the number of cycles, and the longer the homogenization time. Preferably, the time of high pressure homogenization is 4.5-52 min, for example, 4.5 min, 9 min, 18 min, 27 min, 36 min, 40 min, 45 min, and 52 min.

[0026] In the present application, the single time of high pressure homogenization cycle is 1.5 min.

[0027] In the present application, the β-lactamase inhibitor can be a compound as shown in (A).

[0028] In the present application, the pH value in the suspension is the conventional pH value in the art, which can be 4-5, for example, 4, 4.2, 4.5, 4.6, 4.8, or 5.0.

[0029] In the present application, the temperature of high pressure homogenization is the conventional temperature in the art, which can be less than or equal to 55℃, for example, 20℃, 27℃, 30℃, 33℃, 35℃, 38℃, 40℃, 48℃, 50℃, or 51℃.

[0030] In the present application, the particle size of the β-lactamase inhibitor is the conventional particle size in the art, which can be less than or equal to 500 μm, for example, 10-200 μm, for example, 60-100 μm, for example, 31.78 μm, 60.33 μm, or 100 μm.

[0031] In the present application, the suspension further comprises sulfobutylether-beta-cyclodextrin sodium, citric acid and a base. The base is preferably NaOH.

[0032] In the present application, the content of the liquid formulation of the beta-lactamase inhibitor in the suspension is a conventional content in the art, for example, less than or equal to 15 mg / mL; for example, 5.26 mg / mL, 10 mg / mL or 12 mg / mL.

[0033] In the present application, the content of the citric acid in the suspension is a conventional content in the art, for example, 0.69-0.85 mg / mL, for example, 0.77 mg / mL.

[0034] In the present application, the content of the sulfobutylether-beta-cyclodextrin sodium in the suspension is a conventional content in the art, for example, 35-60 mg / mL, for example, 50.00 mg / mL.

[0035] In the present application, the content of the base in the suspension is a conventional content in the art, for example, 5.88-7.18 mg / mL, for example, 6.53 mg / mL.

[0036] In the present application, the homogenizing cavity of the microfluidizer is Y-shaped or Z-shaped, preferably Y-shaped.

[0037] In the present application, the preparation process further comprises centrifugation and / or filtration after high-pressure homogenization.

[0038] In the present application, the filter for filtration has a specification of 0.22 μm.

[0039] The present application further provides a use of high-pressure homogenization in the preparation of a liquid formulation of a beta-lactamase inhibitor, and the conditions of the use are as described in the aforementioned preparation process.

[0040] The present application further provides a liquid formulation of a beta-lactamase inhibitor, which is prepared by the aforementioned preparation process.

[0041] The present application further provides a liquid formulation of a beta-lactamase inhibitor for use as a medicament.

[0042] The present application further provides a liquid formulation of a beta-lactamase inhibitor for use in inhibiting beta-lactamase.

[0043] The present application further provides a method for treating bacterial infection, comprising administering to a patient in need a therapeutically effective amount of an antibacterial agent, and administering a therapeutically effective amount of a liquid formulation of a beta-lactamase inhibitor in combination.

[0044] In a certain embodiment, the antibacterial agent is a beta-lactam antibiotic.

[0045] In certain embodiments, the beta-lactamase inhibitor liquid formulation is administered prior to, concurrently with, or after the antibacterial agent.

[0046] In certain embodiments, the beta-lactamase inhibitor liquid formulation is an injection, and the concentration of the beta-lactamase inhibitor is 5 mg / mL.

[0047] The above-mentioned preferred conditions can be combined arbitrarily, i.e. to obtain the preferred embodiments of the present application, without departing from the common general knowledge in the art.

[0048] The reagents and raw materials used in the present application are commercially available.

[0049] The positive progress effect of the present application is that the liquid formulation of the beta-lactamase inhibitor prepared by the preparation process of the present application has no significant change in concentration and total impurity content when placed at 25°C under natural light for 24H. BRIEF DESCRIPTION OF DRAWINGS

[0050] Figure 1 is a process flow diagram of the original liquid formulation.

[0051] Figure 2 is the experimental result of the stability of the liquid prepared in Example 1.

[0052] Figure 3 is the time change curve of the total impurity content (RP-HPLC) of the liquid prepared in Example 1.

[0053] Figure 4 is the preparation of the liquid formulation of Compound A by the high-pressure homogenization process (high-pressure homogenizer) in Example 2.

[0054] Figure 5 is the effect of the high-pressure homogenization process (microfluidic homogenizer) on the solubility of Compound A in Example 3.

[0055] Figure 6 is the stability study of the liquid after homogenization in Example 3.

[0056] Figure 7 is the appearance comparison of Compound A homogenized liquid prepared by Y-type and Z-type homogenization cavities in Example 4.

[0057] Figures 8 and 9 are the effect of the microfluidic homogenization process of different homogenization cavities (Y-type and Z-type) on the solubility of Compound A in Example 4.

[0058] Figures 10 and 11 are the solubility results of Compound A after microfluidic homogenization with different particle sizes / concentrations in Example 5. DETAILED DESCRIPTION

[0059] The present application will be further described by way of examples, but the present application is not limited to the examples. The experimental methods in the following examples are not specified, and are selected according to conventional methods and conditions, or according to the instructions of the goods.

[0060] The meanings of technical terms in the present application are consistent with the common understanding of those skilled in the art, unless otherwise specified. In the present application, "one" or its combination with various quantifiers includes both singular and plural meanings, unless otherwise specified. In the present application, when multiple numerical values, numerical value ranges, or their combinations are given to explain the same parameter or variable, it is equivalent to simultaneously specifically disclosing these numerical values, range end values, and numerical value ranges formed by any combination thereof. In the present application, any numerical value, whether or not with an approximate modifier such as "about", always covers the approximate range that can be understood by those skilled in the art, such as a range of plus or minus 10%, 5%, etc. In this article, any "embodiment" equally refers to and covers the embodiments of the methods and systems of the present application. In the present application, one or more technical features in any embodiment can be freely combined with one or more technical features in any one or more other embodiments, and the resulting embodiment also belongs to the disclosure of the present application. The weight / volume percentage (%) in this article represents the number of grams (g) per 100 milliliters (100 mL). The molar concentration (M, mol / L) represents the number of moles (mol) of solute contained per liter (L) of solution. When the dispersed substance is uniformly dispersed in the dispersion medium in the form of molecules, atoms, or ions (particle diameter d < 1 nm), the system formed is called a true solution. The true solution is dispersed in the form of small molecules or ions, a homogeneous clear solution, the system is stable, also known as a solution, and the particle size in it is <1 nm.

[0061] In the present application, "homogenizer" includes various types of devices with homogenization function known in the art, such as high-pressure homogenizer, microfluidic homogenizer. High-pressure homogenizer is a commonly used device in pharmaceutical, food, and chemical industries, and the selection and use of its type belong to the skill of those skilled in the art.

[0062] In the present application, "homogenization chamber" includes components with various type parameters and different internal structures, which are core components of high-pressure homogenizers, such as Y / Z type homogenization chamber. In the present application, "liquid preparation" refers to a liquid dispersion system composed of a drug or other ingredients dispersed in a liquid solvent in a certain form. "Compound A" refers to one of the compounds of β-lactamase inhibitors (BLI). "API" refers to Active pharmaceutical ingredient, which is any substance or mixture of substances used in the manufacture of a drug product, which has a pharmacological activity or other direct effect or can affect the function or structure of the body, but cannot be directly taken. Generally, after adding excipients and processing, it is made into a directly usable drug. "Placebo" refers to tablets, pills, or injections made of substances with no drug effect and no toxic side effects, such as citric acid and sulfobutyl ether-β-cyclodextrin (SBECD) used in the present application.

[0063] In some embodiments, the high-pressure homogenization process for the liquid preparation of the broad-spectrum antibacterial poorly water-soluble small-molecule compound of the β-lactamase inhibitor type referred to in the present application includes any device, tool or integration thereof that is capable of achieving the fact that a high concentration of the liquid preparation of the broad-spectrum antibacterial poorly water-soluble small-molecule compound of the β-lactamase inhibitor type is achieved.

[0064] The present application will be described in detail by the following exemplary specific embodiments. The following embodiments are only used to help the skilled in the art to better understand the present application. It should be noted that the spirit of the present application and the protection scope of the claims are not limited by the following specific embodiments.

[0065] The material compound A used in the examples was purchased from Chongqing Bioteng Company;

[0066] The structure of compound A is as follows:

[0067] The high-pressure homogenizer (model: NATHOX Lab 3) was purchased from Nathan Technologies Company; the microfluidic homogenizer (model: M-110EH30) and its accessories Y / Z type homogenizing cavity were purchased from Microfluidics Company; the constant-temperature water bath (model: HWCL-3) was purchased from Greatwall Company. The dynamic light scattering instrument (model: Malvern ZEN3600) was purchased from Malvern; the clarity detector (model YB-2) was purchased from Tianda Tianfa Company.

[0068] Reverse phase high performance liquid chromatography (RP-HPLC): chromatographic column (3 μm, 150 mm x 4.6 mm); by Agilent high performance liquid chromatograph (model: 1260); mobile phase: A: 0.05% phosphoric acid: (980 mL H2O: 5 mL methanol: 5 mL acetonitrile), B: 0.05% phosphoric acid: (500 mL methanol: 500 mL acetonitrile); flow rate: 1.0 mL / min; detection wavelength: 210 nm. Diluent: pure water.

[0069] In the following examples, the detection conditions of the relevant substances (RP-HPLC) are shown in Table 1 as follows:

[0070] Table 1:

[0071] Example 1 Preparation process of compound A liquid preparation

[0072] This example aims to study the physicochemical properties of the compound A liquid drug solution prepared by the liquid preparation process, in order to evaluate the effect of the liquid preparation process on the drug solution.

[0073] Homogeneous liquid formulation: 50.00 mg / mL SBECD, 0.77 mg / mL citric acid, 6.53 mg / mL 1M NaOH, Compound A (batch: PS-12502-105-190002-1) feed concentration of 5.26 mg / mL.

[0074] Homogenization process: high pressure homogenizer (Nathan Technologies, model: NATHOX Lab 3) was used; the temperature was not controlled during the homogenization process; freshly prepared Compound A suspension (150 ml) was homogenized at 5 Kpsi for 3 cycles, with a total cycle time of 4.5 min; at 10 Kpsi for 3 cycles, with a total cycle time of 4.5 min; the homogenization pressure was increased to 20 Kpsi for 18 cycles, with a total cycle time of 27 min.

[0075] According to the liquid storage stability study scheme shown in Table 2, the liquid formulation was prepared and stored at 2-8°C and 25°C respectively for 24H. The liquid storage stability test includes appearance, pH value, insoluble particles, concentration and related substances (RP-HPLC). The specific scheme is shown in Table 2 below.

[0076] Table 2 Compound A liquid storage stability study scheme

[0077] X = appearance, pH, insoluble particles, concentration and related substances (RP-HPLC)

[0078] The results of the Compound A liquid formulation Compound A storage stability experiment are shown in Figure 2. The results show that after storage at 2-8°C and 25°C for 24H, there is no significant change in appearance, pH value, insoluble particles, concentration; there is no significant increase in total impurities; the time variation curve of the total impurities of the prepared liquid (RP-HPLC) is shown in Figure 3.

[0079] Example 2 High pressure homogenization process (high pressure homogenizer) preparation of Compound A liquid formulation

[0080] This example uses a high pressure homogenizer of model NATHOX Lab 3 of Nathan Technologies company to develop a high pressure homogenization preparation process for Compound A liquid formulation.

[0081] The homogeneous liquid formulation was selected: 50.00 mg / mL SBECD (sulfobutyl thesine cyclodextrin sodium), 0.77 mg / mL citric acid, 6.53 mg / mL 1M NaOH, Compound A (batch: PS-12502-105-190002-1) feed concentration of 5.26 mg / mL.

[0082] The homogenization process uses a cooling circulating water cooling method to control the homogenization liquid temperature ≤ 50℃; the freshly prepared compound A suspension (150ml) is used with a high-pressure homogenizer, and after a series of homogenization pressures and different homogenization cycles, the sample is taken: 5Kpsi homogenization pressure for 3 times, total cycle time is 4.5min, 10Kpsi homogenization pressure for 3 times, total cycle time is 4.5min, and the homogenization pressure is increased to 20Kpsi for 18 times, the total cycle time is 27min. Part of the sample after homogenization is placed at 70℃ for 45min. The sample is detected under the clarity detector, and the specific high-pressure homogenization process scheme is shown in Table 3 below.

[0083] Table 3 High-pressure homogenization process (high-pressure homogenizer) for preparing compound A liquid preparation

[0084] X = concentration and related substances (RP-HPLC); Y = temperature; () indicates selected

[0085] " / " indicates that the data does not exist

[0086] The results are summarized in Figure 4, and the sample concentration after high-pressure homogenization (homogenization pressure to 20Kpsi, cycle 18 times, cycle time 27min) and high-pressure homogenization (homogenization pressure to 20Kpsi, cycle 18 times, cycle time 27min) and 70℃ incubation for 45min are very similar, which are 5.01mg / mL and 4.98mg / mL respectively; but the comparison of the results found that the former total impurity content is significantly lower. This shows that in order to ensure that the impurity content of the drug solution is at a low level, the compound A liquid preparation process should avoid high-temperature heating links as much as possible.

[0087] Example 3 High-pressure homogenization process (microfluidic homogenizer) for preparing compound A liquid and high-pressure homogenization process evaluation

[0088] This example uses Microfluidics M-110EH type microfluidic homogenizer to develop high-pressure homogenization preparation process of compound A liquid preparation, investigates the influence of this process on the solubility of compound A, and at the same time investigates the stability of the liquid after homogenization; It can also be used to guide and compare the stability difference between the homogenization process and the original liquid preparation (Example 1).

[0089] Select the homogenization liquid formula: 50.00mg / mL SBECD, 0.77mg / mL citric acid, 6.53mg / mL 1M NaOH, and the feed concentration of compound A solution (batch: DP3ES001101901) is 10.00mg / mL.

[0090] High pressure homogenization process: using microfluidizer homogenizer, Y-type homogenization cavity; control the homogenization process temperature ≤ 50℃; the freshly prepared compound A suspension (150ml) was first homogenized at 5Kpsi pressure for 3 cycles, with a cycle time of 4.5min, then homogenized at 10Kpsi pressure for 3 cycles, with a cycle time of 4.5min, and finally the homogenization pressure was increased to 20Kpsi and homogenized for 24 cycles, with a cycle time of 36min. After taking the last homogenization liquid and filtering it through a 0.22μm filter, it was placed under natural light at 2-8℃ and 25℃ respectively for 24H, and the specific scheme is shown in Table 4 below.

[0091] Table 4 Effect of high pressure homogenization process (microfluidizer homogenizer) on solubility of compound A and stability of liquid after homogenization

[0092] Research scheme

[0093] X = pH, DLS (after filtration), concentration and related substances (RP-HPLC);

[0094] Y = sample temperature

[0095] The research results are summarized in Figures 5 and 6. Using a microfluidizer homogenizer, the microfluidizer homogenization temperature was controlled at ≤ 50℃, the compound A feed concentration was increased to 10.00mg / mL, and the compound A concentration increased with increasing homogenization pressure and homogenization cycle number. The concentration increased to 9.32mg / mL when the homogenization pressure reached 20Kpsi and the homogenization cycle was 24 times with a cycle time of 36min. Compared with the 70℃ heating after high pressure homogenization, the total impurity content was significantly reduced. During the homogenization process, the pH of the selected homogenization sample solution did not change significantly; the particle size detection (DLS) results showed that the compound A homogenization liquid filtered through a 0.22μm filter was a true solution.

[0096] The results of the stability experiment of compound A liquid after homogenization showed that the concentration and total impurity content of compound A homogenization liquid filtered through a 0.22μm filter did not change significantly when placed at 5℃ under natural light for 24H; the concentration and total impurity content did not change significantly when placed at 25℃ under natural light for 24H.

[0097] In addition, in this embodiment, the homogenization pressure was 20Kpsi, the homogenization cycle was 24 times, and the cycle time was 36min. Compared with the results of the original liquid preparation of Example 1, the total impurities were significantly reduced by 1.03%; and the total impurities were also significantly reduced by 1.49% when placed at 25℃ under natural light for 24H.

[0098] Example 4 Effect of different homogenization cavities (Y-type and Z-type) microfluidizer homogenization process on solubility of compound A

[0099] This example uses Microfluidics M-110EH microfluidic homogenizer, and on this basis, Y and Z two homogenization cavities are used; the effects of different homogenization cavity process conditions on the solubility of compound A are investigated.

[0100] Homogeneous liquid formula: 50.00 mg / mL SBECD, 0.77 mg / mL citric acid, 6.53 mg / mL 1M NaOH, and compound A with a feed concentration of 12.00 mg / mL (batch: D153-2019208-0029-01).

[0101] High-pressure homogenization process: microfluidic homogenizer; Y and Z type homogenization cavities are used respectively; the homogenization process control temperature is ≤45℃; the freshly prepared compound A suspension (150 ml) is homogenized for 3 cycles at 5Kpsi homogenization pressure, the cycle time is 4.5 min, homogenized for 3 cycles at 20Kpsi pressure, and the homogenization pressure is increased to 25Kpsi for 27 cycles, and the cycle time is 40 min. The specific scheme is shown in Table 5 below.

[0102] Table 5 Effect of microfluidic homogenization process with different homogenization cavities (Y and Z types) on the solubility of compound A

[0103] X = pH, sample temperature, DLS (after filtration), concentration and related substances (RP-HPLC);

[0104] The appearance results are shown in Figure 7. Under the same compound A feed concentration (12.00 mg / mL) and other homogenization conditions, when the homogenized liquid is relatively clear: as shown in Figures 8 and 9, the Z type homogenization cavity needs to be homogenized to 25K-57 (homogenization pressure (psi)-homogenization cycle (times)), while the Y type homogenization cavity only needs to be homogenized to 25K-30 (homogenization pressure (psi)-homogenization cycle (times)).

[0105] The effect of different homogenization cavities (Y and Z types) on the solubility of compound A is shown in Figures 8 and 9. Compared with the Z type homogenization cavity, the concentration of the compound A liquid preparation prepared by the microfluidic high-pressure homogenization process using the Y type homogenization cavity is significantly higher, and the total impurity content is significantly lower. The particle size detection (DLS) results show that the compound A homogenized liquid prepared using the two homogenization cavities is a true solution after filtration through a 0.22 μm filter.

[0106] In summary, for compound A, the microfluidic homogenization process using the Y type homogenization cavity has better homogenization effect.

[0107] Example 5 Solubility of compound A after microfluidic homogenization of different particle sizes

[0108] On the basis of Example 4, using compound A microfluidizer, Y-type homogenization cavity. To illustrate: under the guidance of the optimized homogenization process, the solubility of compound A of different particle sizes changes; At the same time, the solubility of compound A of two different concentrations of 60.33 pm particle size changes after homogenization.

[0109] Homogenized liquid formula: 50.00 mg / mL SBECD, 0.77 mg / mL citric acid, 6.53 mg / mL 1M NaOH, 31.78 pm, 60.33 pm and 100.00 pm three particle size compound A (batch: Z-D153-20190001-01) feed concentration see Table 6.

[0110] High pressure homogenization process: microfluidizer (Microfluidics, model: M-110EH30); using Y-type homogenization cavity homogenization process control temperature ≤45℃; The freshly prepared compound A suspension (150 ml) was homogenized under 5Kpsi homogenization pressure for 3 cycles, with a cycle time of 4.5 min, then under 10Kpsi homogenization pressure for 3 cycles with a cycle time of 4.5 min, and then the homogenization pressure was increased to 20Kpsi and homogenized for 35 cycles with a cycle time of 52 min. The specific scheme is shown in Table 6 below.

[0111] Table 6 Effect of microfluidization process on solubility of compound A of different particle size / concentration

[0112] X = pH, concentration and related substances (RP-HPLC);

[0113] Y = temperature () indicates selected;

[0114] The effect of microfluidization process on the solubility of compound A of different particle size / concentration is shown in Figures 10 and 11. Under the premise of consistent microfluidization process (i.e. the feed concentration is 5.30 mg / mL; the homogenization pressure and the number of homogenization cycles are consistent; Y-type homogenization cavity is used; the homogenization temperature is controlled at ≤45℃), the pH of the homogenized liquid of compound A of three different particle sizes (31.78 pm, 60.33 pm and 100.00 pm) does not change significantly, and the concentration of compound A is 4.55 mg / mL after homogenization to 20K-35 (homogenization pressure (psi)-homogenization cycle (times)) and filtration through a 0.22 pm filter; indicating that different particle sizes in the range of 31-100 pm have little effect on the solubility of compound A. In combination with the impurity results, the total impurity content of the homogenized liquid of compound A with a particle size of 60.33 pm is relatively low.

[0115] The 60.33 μm raw material was used to prepare the liquid preparation of compound A, and the feeding concentration was increased to 10 mg / mL. The microfluidization homogenization process (temperature control ≤45°C) was used to achieve a solubility of compound A ≥5 mg / mL. When the 60.33 μm raw material was used to prepare the liquid preparation of compound A, the feeding concentration was 5.30 mg / mL, the homogenization was performed for 35 cycles at 20 Kpsi, and the cycle time was 52 min, the solubility of compound A was 4.55 mg / mL. When the feeding concentration was 10.00 mg / mL, the same homogenization condition, the solubility of compound A was 5.21 mg / mL.

[0116] The above merely is the specific application example of the present application, and does not constitute any limitation to the protection scope of the present application. Based on the above description, other different forms of changes or variations can also be made by those skilled in the art in the field. Here, it is not necessary and also impossible to list all the embodiments. Any similar technical solution formed by equivalent transformation or equivalent replacement falls within the protection scope of the present application.

Claims

1. A process for the preparation of a liquid formulation of a β-lactamase inhibitor, characterized in that, The suspension comprising a β-lactamase inhibitor is prepared into a liquid preparation of β-lactamase inhibitor by high pressure homogenization, The pressure of the high pressure homogenization is 1-30K psi; The cycle of the high pressure homogenization is 3-57 times; The equipment of the high pressure homogenization is high pressure homogenizer or microfluidizer.

2. The preparation process according to claim 1, wherein, It meets one or more of the following conditions: (1) The flow rate of the high pressure homogenization is 50-200 ml / min, (2) The pressure of the high pressure homogenization is 5-25K psi, (3) The cycle of the high pressure homogenization is 3 times, 18 times, 24 times, 27 times, 30 times, 35 times or 57 times; (4) The particle size of the β-lactamase inhibitor is 10-200 μm.

3. The preparation process of claim 1, wherein, It meets one or more of the following conditions: (1) The flow rate of the high pressure homogenization is 100 ml / min; (2) The pressure of the high pressure homogenization is 5K psi, 10K psi, 20K psi or 25K psi; (3) The particle size of the β-lactamase inhibitor is 60-100 μm.

4. The preparation process of claim 1, wherein, It meets one or more of the following: (1) When the pressure of the high pressure homogenization is 5K psi, the cycle of the high pressure homogenization is 3 times; (2) When the pressure of the high pressure homogenization is 10K psi, the cycle of the high pressure homogenization is 3 times; (3) When the pressure of the high pressure homogenization is 20K psi, the cycle of the high pressure homogenization is 3 times or 18 times; (4) When the pressure of the high pressure homogenization is 20K psi, the cycle of the high pressure homogenization is 24 or 35 times; (5) When the pressure of the high pressure homogenization is 25K psi, the cycle of the high pressure homogenization is 27, 30 or 57 times; (6) The particle size of the β-lactamase inhibitor is 31.78 μm, 60.33 μm or 100 μm.

5. The preparation process of claim 1, wherein, It meets one or more of the following: (1) The cycle of the high pressure homogenization is 3 times at 5K psi homogenization pressure, 3 times at 10K psi homogenization pressure, and then the homogenization pressure is increased to 20K psi for 35 times of high pressure homogenization; (2) The cycle of the high pressure homogenization is 3 times at 5K psi homogenization pressure, 3 times at 20K psi homogenization pressure, and then the homogenization pressure is increased to 25K psi for 27 times of high pressure homogenization; (3) The cycle of the high pressure homogenization is 3 times at 5K psi homogenization pressure, 3 times at 10K psi homogenization pressure, and then the homogenization pressure is increased to 20K psi for 24 times of high pressure homogenization; (4) The cycle of the high pressure homogenization is 3 times at 5K psi homogenization pressure, 3 times at 10K psi homogenization pressure, and then the homogenization pressure is increased to 20K psi for 18 times of high pressure homogenization.

6. The preparation process of claim 1, wherein, It meets one or more of the following conditions: (1) The time of the high pressure homogenization is 4.5-52 min; (2) The single time of the cycle of the high pressure homogenization is 1.5 min; (3) the beta-lactamase inhibitor is a compound as shown in (A), (4) The pH value in the suspension is 4-5; (5) The temperature of the high pressure homogenization is less than or equal to 55℃; (6) the particle size of the beta-lactamase inhibitor is less than or equal to 500 μm; (7) the suspension further comprises sulfobutyl ether beta-cyclodextrin sodium, citric acid and a base; (8) the homogenization cavity of the microfluidizer is Y-shaped or Z-shaped; (9) in the preparation process, the high pressure homogenization is followed by centrifugation and / or filtration.

7. The preparation process according to claim 6, wherein the step of mixing the first and second components is performed at a temperature of 20 to 30°C. which satisfies one or more of the following conditions: (1) the time of the high pressure homogenization is 4.5 min, 27 min, 36 min, 40 min and 52 min; (2) the pH value of the suspension is 4.5 or 4.6; (3) the temperature of the high pressure homogenization is 20℃, 27℃, 30℃, 33℃, 35℃, 38℃, 40℃, 48℃, 50℃ or 51℃; (4) the homogenization cavity of the microfluidizer is Y-shaped.

8. The preparation process of claim 6, wherein, which satisfies one or more of the following conditions: (1) in the suspension, the base is NaOH; (2) in the suspension, the content of the citric acid is 0.77 mg / mL; (3) in the suspension, the content of the sulfobutyl ether beta-cyclodextrin sodium is 50.00 mg / mL; (4) in the suspension, the content of the base is 6.53 mg / mL; (5) in the preparation process, the specification of the filter for the filtration is 0.22 μm.

9. Use of high pressure homogenization for the preparation of a liquid formulation of a β-lactamase inhibitor, characterized in that, The application satisfies any one of the conditions of claims 1-8.

10. A liquid formulation of a β-lactamase inhibitor characterized in that, The liquid preparation is prepared by the preparation process of any one of claims 1-8.

Images