Purification method of Pinefibrate and Pinefibrate tablet

Abstract

The invention relates to the technical field of oral non-biological medicine preparations, in particular to a purification method of Pinefibrate and a Pinefibrate tablet. In order to solve the problem that impurities in a Pinefibrate tablet can influence the purity and quality of Pinefibrate, the Pinefibrate purification method comprises the following steps: injecting a Pinefibrate solution to be purified into a high performance liquid chromatograph, and then carrying out gradient elution under the chromatographic conditions that disodium hydrogen phosphate and methanol are used as mobile phases and the detection wavelength is 280-290nm, so as to obtain the Pinefibrate. And carrying out gradient elution to obtain the Pinefibrate.

Description

Technical Field

[0001] This application belongs to the field of oral non-biological drug formulation technology, specifically relating to a purification method for pemafibrate and pemafibrate tablets. Background Technology

[0002] Pemafibrate, also known as permabetal, is the first highly selective peroxisome proliferator-activated receptor alpha (PPARα) developed based on a novel concept. For patients with limited efficacy of existing fibrates (including those taking statins and those with impaired renal function or hepatic steatosis), it may offer a better risk-benefit balance than existing fibrates and is considered a safer drug. Its structural formula is as follows:

[0003] However, impurities introduced during the preparation of pemafibrate can affect its quality and safety. These impurities include those introduced from the raw materials used in pemafibrate preparation, impurities generated during the preparation process, and degradation-related impurities. Especially 3-((benzo[d]oxazol-2-yl(3-(4-methoxyphenoxy)propyl)amino)methyl)phenol and (R)-2-(3-((benzo[d]oxazol-2-yl(3-(4-methoxyphenoxy)propyl)amino)methyl)phenoxy)butyrate), wherein 3-((benzo[d]oxazol-2-yl(3-(4-methoxyphenoxy)propyl)amino)methyl)phenol, designated as impurity I, has the following structural formula:

[0004] (R)-2-(3-((benzoxazol-2-yl(3-(4-methoxyphenoxy)propyl)amino)methyl)phenoxy)butyrate butyl ester is designated as impurity II, and its structural formula is:

[0005] These impurities can directly affect the purity and quality of pemafibrate, and may even cause serious adverse reactions, affecting the effectiveness and safety of clinical use. Summary of the Invention

[0006] In view of the technical problems existing in the prior art, this application provides a method for purifying pemafibrate and pemafibrate tablets.

[0007] Firstly, this application provides a method for purifying pemafibrate, which employs the following technical solution: A method for purifying pemafibrate. The method comprises: injecting the pemafibrate solution to be purified into a high-performance liquid chromatograph, and then performing gradient elution to obtain pemafibrate under chromatographic conditions with disodium hydrogen phosphate and methanol as the mobile phase and a detection wavelength of 280 nm-290 nm.

[0008] By employing the above technical solution, based on high-performance liquid chromatography (HPLC), the mobile phase ratio can be dynamically adjusted, effectively separating compounds of different polarities and improving purification efficiency. Furthermore, by using a detection wavelength in the range of 280nm-290nm, specific selection of impurities can be achieved, further reducing interference from impurities. Furthermore, the concentration range of the pemafibrate solution to be purified is 0.5 mg / mL to 2.0 mg / mL.

[0009] By adopting the above technical solution, it can be adapted to high performance liquid chromatography (HPLC) to ensure the separation of the main peak and the impurity baseline, avoid peak overlap, and, in combination with gradient elution, balance the separation effect, reduce residual interference, and further reduce the interference of impurities.

[0010] Furthermore, the solvent for the pemabet solution to be purified is a mixture of methanol and water.

[0011] By adopting the above technical solution, on the one hand, methanol has good solubility for organic compounds such as pemafibrate, and releases less heat when miscible with water, resulting in more uniform mixing of the mobile phase and a more stable baseline. The ultraviolet cutoff wavelength of methanol (210 nm) is suitable for detection wavelengths of 280-290 nm, avoiding baseline noise at low wavelengths, and is also well matched with the detection wavelength of pemafibrate. On the other hand, the methanol and water system produces lower column pressure, which can extend the column life and reduce the instrument load.

[0012] Furthermore, the volume ratio of methanol to water is 8:5.

[0013] By adopting the above technical solution, pemafibrate (lipid-soluble) can be effectively dissolved while retaining sufficient aqueous phase to maintain the polarity gradient of the mobile phase, thus avoiding peak broadening caused by pure organic phase. Moreover, the above methanol and water volume ratio setting is compatible with disodium hydrogen phosphate buffer, which can stably maintain pH value, reduce column efficiency fluctuations, and is also suitable for the analysis of the concentration range of the pemafibrate solution to be purified.

[0014] Furthermore, the concentration of disodium hydrogen phosphate is 0.01 mol / L.

[0015] Furthermore, the pH of the mobile phase is 4.0.

[0016] By employing the above technical solution, a disodium hydrogen phosphate concentration of 0.01 mol / L provides sufficient buffering capacity while avoiding excessively high salt concentrations that could lead to increased column pressure or instrument clogging. A mobile phase pH of 4.0 reduces secondary interactions between basic analytes and silanol groups, improving peak shape (e.g., reducing tailing). Furthermore, the aforementioned disodium hydrogen phosphate concentration and mobile phase pH settings are compatible with gradient elution, suitable for pemafibrate solutions with concentrations ranging from 0.5 mg / mL to 2.0 mg / mL, facilitating the separation of impurities.

[0017] Furthermore, in a high-performance liquid chromatograph, the column temperature is 30℃-40℃.

[0018] By employing the above technical solutions, mass transfer rate and longitudinal diffusion can be balanced, peak broadening can be reduced, column efficiency can be improved, retention time drift caused by temperature fluctuations can be avoided, and the reproducibility of analytical results can be ensured. Furthermore, the column packing material is octadecyl bonded silica gel.

[0019] By adopting the above technical solution, the hydrophobic interaction of octadecyl bonded silica gel can more easily separate pemabet and impurities, making it easier to separate the impurities.

[0020] Furthermore, the inner diameter of the chromatographic column is 4.6 mm, the length of the chromatographic column is 150 mm, and the diameter of the stationary phase particles in the chromatographic column is 3.0 μm, that is, the specifications are 4.6 mm × 150 mm and 3 μm.

[0021] By adopting the above technical solution, the inner diameter of the chromatographic column is 4.6 mm, the length of the chromatographic column is 150 mm, and the diameter of the stationary phase particles of the chromatographic column is 3.0 μm. This ensures sufficient sensitivity (suitable for the concentration range of the pemafibrate solution to be purified from 0.5 mg / mL to 2.0 mg / mL), making it easier to separate pemafibrate from impurities, and making it easier to separate the impurities.

[0022] Furthermore, the flow rate of the mobile phase ranges from 0.8 mL / min to 1.2 mL / min.

[0023] By adopting the above technical solution Secondly, this application provides a pemafibrate tablet. This pemafibrate tablet is prepared from pemafibrate obtained by the aforementioned purification method.

[0024] Understandably, the beneficial effects of the pemafibrate tablets provided in this application are the same as those of the above-mentioned pemafibrate purification method, and will not be repeated here.

[0025] In summary, this application includes at least one of the following beneficial technical effects: (1) The purification method of pemabet in this application can dynamically adjust the mobile phase ratio based on high performance liquid chromatography, effectively separate compounds of different polarities, improve purification efficiency, and specifically select impurities by detecting wavelengths in the range of 280nm-290nm, further reducing the interference of impurities. (2) This application uses disodium hydrogen phosphate and methanol as the mobile phase, and the concentration of disodium hydrogen phosphate of 0.01 mol / L can provide sufficient buffering capacity while avoiding excessive salt concentration that could lead to column pressure increase or instrument blockage. The pH of the mobile phase is 4.0, which can reduce the secondary interaction between the basic analyte and the silanol group and improve the peak shape (such as reducing tailing). Moreover, the above-mentioned concentration of disodium hydrogen phosphate and the pH setting of the mobile phase are compatible with gradient elution and are suitable for the concentration range of the pemafibrate solution to be purified from 0.5 mg / mL to 2.0 mg / mL, making it easier to separate impurities; (3) The purification method of pemabet provided in this application adopts a novel buffer system, which reduces the collection time, reduces the cost of the analysis process, and is fast, simple, accurate and reproducible. It is suitable for the control of impurities and stability studies, and lays the foundation for the formulation of impurity quality standards. Detailed Implementation

[0026] The technical solutions in some embodiments of this application are clearly and completely described below with reference to specific examples. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments provided in this application are within the scope of protection of this application.

[0027] Unless the context otherwise requires, throughout the specification and claims, the term "comprising" is interpreted as open-ended and encompassing, meaning "including, but not limited to." In the description of the specification, terms such as "one embodiment," "some embodiments," "exemplary embodiment," "exemplary," or "some examples," etc., are intended to indicate that a particular feature, structure, material, or characteristic associated with that embodiment or example is included in at least one embodiment or example of this application. The illustrative representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, a particular feature, material, or characteristic may be included in any suitable manner in any one or more embodiments or examples.

[0028] Hereinafter, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature.

[0029] Unless otherwise defined, all technical terms used herein have the same meaning as commonly known to those skilled in the art. In this application, "at least one" means one or more, and "more than one" means two or more. "At least one of the following" or similar expressions refer to any combination of these items, including any combination of single or multiple items. For example, at least one of a, b, or c can mean: a, b, c, a and b, a and c, b and c, or a, b, and c, where a, b, and c can be single or multiple.

[0030] All the following raw materials mentioned in this application are commercially available products and are intended to fully disclose the raw materials in this application. They should not be construed as limiting the source of the raw materials.

[0031] Table 1 Selection of Chromatographic Columns Serial Number Manufacturer filler Specification 1 CORTECS T3 Octadecyl bonded silica gel 4.6mm × 150mm, 3μm 2 <![CDATA[Waters XBridge ® ]]> Octadecyl bonded silica gel 4.6mm×150mm, 5μm 3 Moonrise Octadecyl bonded silica gel 4.6mm × 150mm, 3μm 4 Agilent Octadecyl bonded silica gel 4.6mm × 150mm, 3μm Example 1 Example 1 provides a method for purifying pemafibrate, which includes the following steps: (1) Sample preparation: Take pemafibrate, impurity I and impurity II, dilute with a mixture of methanol and water in a volume ratio of 8:5 to prepare a solution with a concentration of 1 mg / mL. Each 1 mL of this solution contains approximately 1 mg of pemafibrate and 10 μg each of impurity I and impurity II. The chromatographic column is selected as octadecylsilane bonded silica gel as the packing material (the specifications are 4.6 mm × 150 mm, 5 μm or equivalent chromatographic column, i.e. column No. 2 in grid 1).

[0032] (2) A Thermo UltiMate 3000 liquid chromatograph with a UV variable wavelength detector was selected. 0.01 mol / L disodium hydrogen phosphate (pH adjusted to 4.0 with phosphoric acid) and methanol were used as the mobile phase. Under the conditions of column temperature of 35℃ and detection wavelength of 285 nm, gradient elution was performed with an injection volume of 10 μL at a flow rate of 1 mL / min. The specific gradient elution is shown in Table 2 below.

[0033] Table 2 Gradient elution Time / min Disodium hydrogen phosphate / % Methanol / % 0 95 5 10 60 40 15 40 60 19 20 80 29 20 80 30 95 5 35 95 5 Examples 2-4 Examples 2-4 provide a method for purifying pemafibrate, which is basically the same as that in Example 1, except that in step (3), the chromatographic columns are selected from columns 1, 3 and 4 in Table 1 respectively.

[0034] Examples 5-6 Examples 5-6 provide a method for purifying pemafibrate, which is basically the same as that in Example 1, except that the column temperature in step (3) is 33°C and 37°C. The separation degree between impurities and pemafibrate was examined.

[0035] Examples 7-8 Examples 7-8 provide a method for purifying pemafibrate, which is basically the same as that in Example 1, except that the flow rate in step (3) is 0.9 mL / min and 1.1 mL / min, respectively.

[0036] Examples 9-10 Examples 9-10 provide a method for purifying pemafibrate, which is basically the same as that in Example 1, except that the pH of the mobile phase in step (3) is 2.8 and 3.0, respectively.

[0037] Performance testing The resolution and tailing factor of the above examples were determined using liquid chromatography, and the results are as follows.

[0038] Result 1: The test results for Examples 1-4 show that the resolution between the peaks of each component in the general applicability solution of Examples 1-4 is ≥5.0, with the resolution of Example 1 being 6.4, Example 2 being 6.5, Example 3 being 6.0, and Example 4 being 6.2; and the tailing factor is ≤1.2 for all examples, with the tailing factor of Example 1 being 0.94, Example 2 being 0.85, Example 3 being 0.83, and Example 4 being 0.80. The results meet the requirements, indicating that using different brands and models of chromatographic columns has virtually no impact on the detection results.

[0039] Result 2: The separation degree of impurities and pemafibrate in the purification methods of pemafibrate provided in Examples 5-6; the results are shown in Table 3 below.

[0040] Table 3. Durability test results (column temperature change)

[0041] Here, the system suitability solution is prepared by diluting pemafibrate, impurity I and impurity II with a mixture of methanol and water in a volume ratio of 8:5 to prepare a solution with a concentration of 1 mg / mL. Each 1 mL of this solution contains approximately 1 mg of pemafibrate and 10 μg each of impurity I and impurity II.

[0042] It can be seen that under the condition of column temperature change of ±2℃, the resolution between the peaks of each component in the system suitability solution is ≥6.4, and the tailing factor is ≤1.07. The results meet the requirements, indicating that the column temperature change of ±2℃ has basically no impact on the detection results.

[0043] Result 3: The separation degree of impurities and pemafibrate in the purification methods of pemafibrate provided in Examples 7-8 is shown in Table 4 below.

[0044] Table 4 Durability Test Results

[0045] Here, the system suitability solution is prepared by diluting pemafibrate, impurity I and impurity II with a mixture of methanol and water in a volume ratio of 8:5 to prepare a solution with a concentration of 1 mg / mL. Each 1 mL of this solution contains approximately 1 mg of pemafibrate and 10 μg each of impurity I and impurity II.

[0046] It can be seen that, under the condition of a flow rate change of ±0.1 mL / min, the resolution between the peaks of each component in the system suitability solution is ≥6.4, and the tailing factor is ≤1.07. The results meet the requirements, indicating that the flow rate change of ±0.1 mL / min has basically no effect on the detection results.

[0047] Result 4: The separation degree of impurities and pemafibrate in the purification methods of pemafibrate provided in Examples 9-10 is shown in Table 5 below.

[0048] Table 5 Durability Test Results

[0049] Here, the system suitability solution is prepared by diluting pemafibrate, impurity I and impurity II with a mixture of methanol and water in a volume ratio of 8:5 to prepare a solution with a concentration of 1 mg / mL. Each 1 mL of this solution contains approximately 1 mg of pemafibrate and 10 μg each of impurity I and impurity II.

[0050] It can be seen that under the condition of a mobile phase pH change of ±0.2, the resolution between the peaks of each component in the system suitability solution is ≥5.9, and the tailing factor is ≤1.05. The results meet the requirements, indicating that a mobile phase pH change of ±0.2 has virtually no impact on the detection results.

[0051] In summary, the purification method for pemabet provided in this application uses a self-comparison method to calculate the content of related substances, and with reference to the impurity limit, the concentration of the reference standard is 0.1% of the concentration of the test sample, i.e., 1 μg / mL.

[0052] The purification method for pemafibrate described in this application is used for the efficient separation and detection of pemafibrate and its related substances in pemafibrate preparations. This impurity analysis and detection method achieves baseline separation of various impurities in pemafibrate, with good peak shapes, moderate analysis time, and good separation, meeting the analytical requirements for pemafibrate impurities. This pemafibrate purification method is rapid, simple, accurate, and reproducible, and can effectively analyze related substances in pemafibrate, providing possibilities for the quality control of pemafibrate and the safety of drug use.

[0053] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A method for purifying pemafid, characterized in that, The pemafibrate solution to be purified was injected into a high-performance liquid chromatograph, and then gradient elution was performed under chromatographic conditions with disodium hydrogen phosphate and methanol as the mobile phase and a detection wavelength of 280 nm-290 nm to obtain the pemafibrate.

2. The method for purifying pemafidate according to claim 1, characterized in that, The concentration range of the pemafibrate solution to be purified is 0.5 mg / mL to 2.0 mg / mL.

3. The method for purifying pemafidate according to claim 1, characterized in that, The solvent for the pemabet solution to be purified is a mixture of methanol and water.

4. The method for purifying pemafidate according to claim 3, characterized in that, The volume ratio of methanol to water is 8:

5.

5. The method for purifying pemafidate according to claim 4, characterized in that, The concentration of the disodium hydrogen phosphate is 0.01 mol / L; and / or the pH of the mobile phase is 4.

0.

6. The method for purifying pemafidate according to claim 5, characterized in that, In the high-performance liquid chromatograph, the column temperature is 30℃-40℃.

7. The method for purifying pemafidate according to any one of claims 1-6, characterized in that, The column packing material is octadecyl bonded silica gel.

8. The method for purifying pemafidate according to claim 7, characterized in that, The chromatographic column has an inner diameter of 4.6 mm, a length of 150 mm, and a stationary phase particle diameter of 3.0 μm.

9. The method for purifying pemafidate according to any one of claims 1-8, characterized in that, The flow rate of the mobile phase is in the range of 0.8 mL / min to 1.2 mL / min.

10. A pemafibrate tablet prepared by the purification method of pemafibrate as described in any one of claims 1-9.

Images