Method for determining dissolution rate of cabozantinib analogue solid preparation

Abstract

The invention discloses a method for determining the dissolution rate of a cabozantinib analogue solid preparation. The method comprises the following steps: taking a hydrochloric acid medium with the pH value of 1.0 as a dissolution medium; the method comprises the following steps: dissolving cabozantinib analogues in a solvent, adopting a paddle method at the rotating speed of 40-60 revolutions per minute, filtering a dissolution solution after 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes and 60 minutes, taking a continuous filtrate as a test solution, determining the content of the cabozantinib analogues in the test solution by adopting a high performance liquid chromatograph, comparing the content with a labeled amount, and calculating the dissolution rate of the cabozantinib analogues. The determination method provided by the invention can be used for evaluating the quality of samples among batches of preparations, provides indexes for research and development of new drug preparations, provides guarantee for optimization of a preparation process, and ensures the consistency of drug quality and curative effect.

Description

Technical Field

[0001] The present invention relates to the field of drug analysis and detection, and in particular to a method for determining the dissolution rate of a solid preparation of a cabozantinib analog. Background Art

[0002] Dissolution is an important indicator for quality control of solid preparations. As a Class 1 new drug, cabozantinib analogs have not been included in the pharmacopoeias and patent literature of various countries in the world for the determination of the dissolution of solid preparations of cabozantinib analogs. Therefore, it is necessary to establish an accurate and effective detection and analysis method for solid preparations of cabozantinib analogs to objectively reflect the dissolution of cabozantinib analogs in solid preparations, and provide a basis for the quality control and scientific and safe use of cabozantinib analog tablets. Summary of the invention

[0003] The chemical name of the cabozantinib analogue of the present invention is (S)-2-(4-(7-(2-fluoro-4-(1-((4-fluorophenyl)carbamoyl)cyclopropanecarboxamide)phenoxy)thieno[3,2-b]pyridin-2-yl)-1H-pyrazol-1-yl)ethanol valine ester dihydrochloride monohydrate, and its chemical formula is C 34 H 32 F2N6O5S·2HCl·H2O, the chemical structure is shown in (I), and the molecular weight is 765.65.

[0004]

[0005] The purpose of the present invention is to provide a method for determining the dissolution rate of a solid preparation of a cabozantinib analog, which is used to evaluate the consistency of quality between preparation batches and provide an indicator for the research and development of innovative drug preparations. It can be used as an indicator to confirm the consistency of drug quality and efficacy after product stability inspection and some changes (such as prescription, production process, production site changes and production process scale-up).

[0006] The technical solution of the present invention is:

[0007] Using pH 1.0 hydrochloric acid solution as the dissolution medium, the paddle method was adopted at a rotation speed of 40 to 60 revolutions per minute, the dissolution solution was filtered at different time points, and the filtrate was taken as the test solution. The content of cabozantinib analogs in the test solution was then determined, and then compared with the labeled amount of cabozantinib analogs of the solid preparation of cabozantinib analogs to calculate its solubility.

[0008] Preferably, the rotation speed adopted in the present invention is 50 revolutions per minute.

[0009] Preferably, the time for taking the dissolution solution in the present invention is 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes and 60 minutes respectively.

[0010] Preferably, the preparation method of the dissolution medium of the present invention, pH 1.0 hydrochloric acid, is as follows: take 9 ml of concentrated hydrochloric acid, dilute to 1000 ml with water, and shake well.

[0011] Preferably, the preparation method of the reference solution of the present invention is: accurately measure an appropriate amount of the cabozantinib analog reference substance and place it in a volumetric bottle, add acetonitrile-water (70:30) to dissolve and dilute to the scale, shake well, accurately measure an appropriate amount, dilute with the corresponding pH 1.0 dissolution medium to make a solution containing approximately 27.8 μg of cabozantinib analogs per 1 ml, shake well, and use it as the reference solution.

[0012] Preferably, the present invention uses high performance liquid chromatography to determine the content of the cabozantinib analog in the test solution.

[0013] Preferably, the solid preparation of cabozantinib analogs of the present invention is a cabozantinib analog tablet.

[0014] Preferably, the tablets of the present invention are prepared according to Chinese patent application ZL202210358212.1, that is, the cabozantinib analogs are uniformly mixed with excipients, and then tabletted and coated.

[0015] Preferably, the specific steps of the method of the present invention are:

[0016] (1) Prepare the dissolution medium of hydrochloric acid at pH 1.0: Take 9 ml of concentrated hydrochloric acid, dilute to 1000 ml with water, and shake well;

[0017] (2) preparing a test solution: dissolving the tablet in the dissolution medium obtained in step (1) by a slurry method at a rotation speed of 40-60 revolutions per minute, taking the dissolution solution at 5, 10, 15, 20, 30, 45, and 60 minutes, respectively, filtering through a microporous filter membrane, and taking each subsequent filtrate as the test solution;

[0018] (3) Preparation of reference solution: accurately weigh an appropriate amount of cabozantinib analog reference substance and place it in a volumetric bottle, add acetonitrile-water (70:30) to dissolve and dilute to the scale, shake well, accurately measure an appropriate amount, and then dilute with the dissolution medium to make a solution containing approximately 27.8 μg of cabozantinib analog per 1 ml, shake well, and use this as the reference solution;

[0019] (4) Determination of dissolution: 10 μl of the reference solution and each of the test solution were injected into a high performance liquid chromatograph and the chromatogram was recorded. The dissolution at different time points was calculated by peak area according to the external standard method.

[0020] Preferably, the high performance liquid chromatography conditions used in the present invention are: the chromatographic column is an octadecylsilane bonded silica gel column, the mobile phase is a potassium dihydrogen phosphate solution-acetonitrile with a volume ratio of 30:70 and a pH value of 6.0, isocratic elution, the concentration of the potassium dihydrogen phosphate solution is 0.01 mol / L, the column temperature of the chromatographic column is 40°C, the flow rate is 1.5 ml / min, and the detection wavelength is 252 nm.

[0021] Preferably, the chromatographic column selected in the present invention is an Agilent Poroshell 120 BonusRP chromatographic column.

[0022] The high performance liquid chromatography method adopted in the present invention realizes an extremely short testing time (the time for testing one injection) and can separate the main component (i.e. the raw material drug cabozantinib analogue) from all other impurities, and quickly and accurately quantitatively detect the content of the main component.

[0023] The present invention adopts the above technical solution and has the following technical effects:

[0024] The cabozantinib analogs of the present invention belong to Class 1 new drugs, and the relevant dissolution determination methods thereof are not currently included in the pharmacopoeias and patent documents of various countries in the world. The present invention provides a new method for determining the dissolution of cabozantinib analog solid preparations, which is of great significance to the quality control and scientific use of cabozantinib analog tablets. The determination method of the present invention can quickly and accurately determine the dissolution of cabozantinib analog solid preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The technical scheme of the present invention will be described in detail below through examples and drawings, so as to better understand the advantages of the present invention. The provided examples should be interpreted as an illustration of the method of the present invention, and do not limit the technical scheme and protection scope disclosed by the present invention in any way.

[0026] Figure 1 is a high performance liquid chromatogram of a cabozantinib analog reference solution;

[0027] Figure 2 is the HPLC chromatogram of the test solution;

[0028] Figure 3 is the HPLC chromatogram of the blank pH 1.0 dissolution medium of Example 3;

[0029] Figure 4 It is the HPLC chromatogram of the blank auxiliary material solution of Example 3;

[0030] Figure 5 Schematic diagram of the dissolution curves of the cabozantinib analog tablets measured when the three batches of self-developed preparations provided in Example 2 were in a hydrochloric acid medium at pH 1.0;

[0031] Figure 6 This is the linear graph of Example 6. DETAILED DESCRIPTION

[0032] The following will be combined with the accompanying drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of the characteristics and implementation schemes of a part of the embodiments of the present invention, rather than all the embodiments. Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention.

[0033] In the following specific embodiments, the reference substances, samples, reagents and instruments used are:

[0034] Instruments: SOTAX XTend series intelligent dissolution apparatus; Agilent 1260 series high performance liquid chromatograph;

[0035] Reagents: hydrochloric acid (analytical grade), potassium hydroxide (analytical grade), acetonitrile (chromatographic grade), potassium dihydrogen phosphate (analytical grade).

[0036] Chromatographic column:

[0037] Agilent Poroshell 120BonusRP, 4.6mm×150mm, 2.7μm;

[0038] The solvent mentioned in the present invention, unless otherwise specified, is ultrapure water.

[0039] Cabozantinib analogs were purchased from the market or prepared according to the method of Chinese patent application ZL202010099747.2 (compound code (td32-4P10)); batch number: J20220802, content: 99.1%;

[0040] Cabozantinib analog tablets are prepared by the preparation room of Shenzhen Neptunus Pharmaceutical Technology Research Institute Co., Ltd., main ingredient: cabozantinib analog, specification: 25 mg, batch number: 20230101, containing cabozantinib analog (C 34 H 32 F2N6O5S·2HCl) is 102.8% of the labeled amount. The preparation method is to mix the cabozantinib analogue with the auxiliary materials uniformly, press the tablets and coat the tablets according to Chinese patent application ZL202210358212.1.

[0041] Cabozantinib analog reference substance: purchased from commercial suppliers or prepared according to the method of Chinese patent application ZL202010099747.2 (compound code (td32-4P10)), batch number: 20221201, content of 96.5%.

[0042] [Example 1] Determination of Cabozantinib Analogs by High Performance Liquid Chromatography

[0043] 1.1 Solution preparation

[0044] Intermediate stock solution: Take about 35 mg of each of cabozantinib analog intermediate 1, cabozantinib analog intermediate 2 and cabozantinib analog intermediate 3 (for detailed information on the intermediates, see Patent No.: ZL202111625993.8), accurately weigh, place in a 25 ml volumetric flask, add about 4 ml of tetrahydrofuran to dissolve, dilute to the scale with acetonitrile-water (70:30), shake well, and obtain.

[0045]

[0046] Preparation of mixed solution: Take about 35 mg of cabozantinib analog reference substance, accurately weigh, place in a 25 ml volumetric flask, add acetonitrile-water (70:30) to dissolve and dilute to the scale as the reference substance stock solution; accurately measure 1 ml of the reference substance stock solution and 10 μl of the intermediate stock solution, place in a 50 ml volumetric flask, dilute to the scale with water, shake well, and obtain.

[0047] 1.2 Selection of chromatographic columns

[0048] On the basis of the determination method of related substances of cabozantinib analogs (patent number: ZL202111625993.8), 0.02 mol / L potassium dihydrogen phosphate buffer (pH 6.0)-acetonitrile (30:70) was used as the mobile phase, the detection wavelength was 252 nm, the column temperature was 30°C, the flow rate was 1.0 ml / min, different chromatographic columns were selected, and the chromatographic column information was shown in Table 1. 10 μl of the mixed solution under item 1.1 was injected into the liquid chromatograph, and the chromatogram was recorded. The main peak retention time, theoretical plate number, tailing factor, and separation between the main peak and adjacent peaks were used as evaluation indicators to screen and optimize the chromatographic column. The results are summarized in Table 2.

[0049] Table 1 Dissolution test method screening - chromatographic column information table

[0050] Serial number Chromatographic columns ① Waters XBridge Shield RP18 (4.6mm×150mm, 3.5μm) ② Waters XBridge C18 (4.6mm×150mm, 3.5μm) ③ Agilent Poroshell 120BonusRP (4.6mm×150mm, 2.7μm) ④ Waters Symmetry C18 (3.9mm×150mm, 5μm)

[0051] Table 2 Dissolution test method screening - chromatographic column selection test results

[0052]

[0053]

[0054] Under the conditions of chromatographic columns ① and ②, the impurities before the main peak did not reach baseline separation with the main peak; under the conditions of chromatographic column ④, the tailing factor of the main peak was large and the number of theoretical plates was too low; under the conditions of chromatographic column ③, the main peak was well separated from the impurities before and after it, and the tailing factor and theoretical plate number were better, so the chromatographic column was tentatively determined to be Agilent Poroshell 120BonusRP (4.6mm×150mm, 2.7μm).

[0055] 1.3 Selection of mobile phase

[0056] Under the premise that other chromatographic conditions remain unchanged, the proportion of acetonitrile in the mobile phase was adjusted, 10 μl of the mixed solution prepared in Example 1.1 was accurately measured and injected into the liquid chromatograph respectively. The main peak elution time, tailing factor, theoretical plate number, separation degree, and maximum retention time of impurities were used as the evaluation indicators for comparison. The results are shown in Table 3.

[0057] Table 3 Dissolution test method screening - selection test results of mobile phase ratio

[0058]

[0059] The results showed that when the mobile phase A-mobile phase B, i.e. 0.02 mol / L potassium dihydrogen phosphate buffer (pH 6.0)-acetonitrile, was adjusted to 25:75, the maximum retention time of the impurity was reduced to 5.088 min, but the separation between the impurity and the main peak was reduced. At the same time, considering that the solubility of potassium dihydrogen phosphate is lower in a high-proportion organic phase, the risk of salting out in a high-proportion organic phase is higher, so the mobile phase A-mobile phase B ratio is tentatively set to (30:70).

[0060] 1.4 Selection of buffer salt concentration

[0061] Under the premise that other chromatographic conditions remain unchanged, different concentrations of buffer salts were used to accurately measure 10 μl of the mixed solution prepared in Example 1.1 and injected into the liquid chromatograph respectively. The main peak elution time, tailing factor, theoretical plate number and separation degree were used as the evaluation indicators for comparison. The results are shown in Table 4.

[0062] Table 4 Dissolution test method screening - buffer salt concentration selection test results

[0063]

[0064] The results showed that reducing the buffer salt concentration from 0.02 mol / L to 0.01 mol / L had no significant effect on the retention time, peak area, theoretical plate number and tailing factor of NEP018, and reduced the risk of salting out of the buffer salt in a high proportion of organic phase. Therefore, the buffer salt concentration of the mobile phase was tentatively set at 0.01 mol / L.

[0065] 1.5 Selection of column temperature

[0066] Under the premise that other chromatographic conditions remain unchanged, different column temperatures were used, and 10 μl of the mixed solution prepared in Example 1.1 was accurately measured and injected into the liquid chromatograph respectively. The main peak elution time, tailing factor, theoretical plate number and separation degree were used as the evaluation indicators for comparison. The results are shown in Table 5.

[0067] Table 5 Dissolution test method screening - chromatographic column temperature selection test results

[0068]

[0069] The results showed that when the column temperature increased from 30℃ to 40℃, the number of theoretical plates increased and the tailing factor decreased, so the column temperature was temporarily set at 40℃.

[0070] 1.6 Selection of mobile phase flow rate

[0071] Under the premise that other chromatographic conditions remain unchanged, different flow rates were used to accurately measure 10 μl of the mixed solution in item 1.1 and inject them into the liquid chromatograph respectively. The main peak elution time, tailing factor, theoretical plate number and maximum retention time of impurities were used as evaluation indicators for comparison. The results are shown in Table 6.

[0072] Table 6 Dissolution test method screening - mobile phase flow rate selection test results

[0073]

[0074] The results showed that when the flow rate was increased to 1.5 ml / min, the main peak elution time and the maximum retention time of impurities decreased to 4.040 min, which could shorten the running time and minimize the main peak tailing factor. Therefore, the flow rate was temporarily set to 1.5 ml / min.

[0075] Therefore, the chromatographic conditions adopted by the present invention are as follows: the chromatographic column is an octadecylsilane bonded silica gel column, the length of the chromatographic column is 150 mm, the inner diameter is 4.6 mm, and the size of the filler particles is 2.7 μm; the mobile phase is 0.01 mol / L potassium dihydrogen phosphate solution (pH value is adjusted to 6.0 with potassium hydroxide)-acetonitrile (30:70) as the mobile phase, isocratic elution; column temperature: 40°C; flow rate is 1.5 ml / min; detection wavelength is 252 nm; solvent: acetonitrile-water (70:30). The high performance liquid chromatography method of the present invention realizes an extremely short test time (the time for testing one injection) and can separate the main component (i.e., the bulk drug cabozantinib analog) from all other impurities, and quickly and accurately quantitatively detect the content of the main component.

[0076] [Example 2] Determination of the dissolution rate of cabozantinib analog tablets

[0077] 2.1 Selection of dissolution curve

[0078] 2.1.1 Selection of dissolution medium

[0079] According to the "Guidelines for Determination and Comparison of Dissolution Curves of Ordinary Oral Solid Preparations" and the "Technical Guidelines for Dissolution Tests of Ordinary Oral Solid Preparations", the dissolution curves of cabozantinib analogs of cabozantinib analog tablets were determined by the paddle method at 50 rpm, using conventional media: 900 ml each of water, pH 1.0 hydrochloric acid solution, pH 4.5 acetate buffer, and pH 6.8 phosphate buffer without surfactant as dissolution media. The results are shown in Table 7.

[0080] Table 7 Dissolution curves of cabozantinib analogs in different dissolution media

[0081]

[0082] The results showed that the dissolution amount of the main component in the pH 1.0 medium was greater than 85% in 20 minutes, and the RSD values ​​of the dissolution amounts at each time point were less than 15%; except for 15 minutes, the dissolution amounts at each time point in the pH 6.8 medium were less than 2%, and the calculable RSDs were greater than 30%, so this medium was not used for subsequent dissolution curve determination; within the cutoff time, the average dissolution amount of NEP018 in water medium and pH 4.5 medium did not reach 85%, and subsequent attempts were made to increase the rotation speed or add an appropriate amount of surfactant to develop the dissolution curves in water medium and pH 4.5 medium.

[0083] 2.1.2 Investigation of surfactant types

[0084] The solubility of this product in water medium and pH4.5 medium at each time point is low, and the RSD of dissolution amount is large. Increasing the rotation speed alone cannot improve the above problems. Therefore, different types and concentrations of surfactants (such as SDS: sodium dodecyl sulfate, CTAB: hexadecyltrimethylammonium bromide) were added to water medium and pH4.5 medium as dissolution medium, paddle method 50rpm, medium volume 900ml, and the types of surfactants added in the dissolution test of tablets containing cabozantinib analogs were investigated to develop the types of surfactants added to the water medium and pH4.5 medium of this product. The results are shown in Table 8.

[0085] Table 8 Surfactant type selection results

[0086]

[0087] The results showed that: ① In the aqueous medium with 0.10% SDS, the dissolution amount of the main component was 58.3% at 90 minutes, and in the aqueous medium with 0.10% CTAB, the dissolution amount of the main component increased to 81.0% at 90 minutes, so CTAB was selected as the surfactant of the aqueous medium; ② In the pH4.5 medium, the dissolution curve of 0.10% SDS added did not change from 5 minutes to 90 minutes (75.1% to 79.3%), which was not discriminatory. The dissolution curve measured in the pH4.5 medium with 0.10% CTAB was more discriminatory, so CTAB was selected as the surfactant of the pH4.5 medium. On this basis, the addition of CTAB in the two media was investigated to see whether increasing the rotation speed could effectively increase the end point dissolution.

[0088] 2.1.3 Investigation of rotation speed

[0089] The dissolution paddle speed test was carried out on tablets containing cabozantinib analogs using pH 1.0 medium, water (containing 0.1% CTAB) medium, and pH 4.5 (containing 0.1% CTAB) medium as dissolution media, paddle speeds of 50 rpm and 75 rpm, and a medium volume of 900 ml to determine the dissolution speed of the product. The test results are shown in Table 9.

[0090] Table 9 Results of speed selection

[0091]

[0092]

[0093] The results showed that the water (containing 0.1% CTAB) medium and pH4.5 (containing 0.1% CTAB) medium, with the increase of the rotation speed, improved the interference of the large RSD of dissolution caused by the stacking of auxiliary materials after the tablet disintegrated, the dissolution amount at the same time point increased, the RSD value of the dissolution amount decreased, and the repeatability between parallel samples was higher, so the paddle method 75rpm was used as the dissolution speed of the water (containing 0.1% CTAB) medium and pH4.5 (containing 0.1% CTAB) medium. However, the end point dissolution was still 85-90% of the labeled amount, not reaching 100%. The pH1.0 medium increased the rotation speed to 75 rpm, and the main component reached 98.8% at 5min, which had no distinguishing power, so the paddle method 50rpm was used as the dissolution speed of the pH1.0 medium.

[0094] 2.2 Finalization of the dissolution test method

[0095] 2.2.1 Preparation of reference solution

[0096] Accurately weigh an appropriate amount of cabozantinib analog reference substance and place it in a volumetric flask, add acetonitrile-water (70:30) to dissolve and dilute to the scale, shake well, accurately measure an appropriate amount, dilute with a dissolution medium pH 1.0 hydrochloric acid solution to make a solution containing approximately 27.8 μg of cabozantinib analog per 1 ml, shake well, and use as the reference substance solution.

[0097] 2.2.2 Test solution

[0098] Take 900 ml of pH 1.0 hydrochloric acid solution (take 9 ml of concentrated hydrochloric acid, dilute to 1000 ml with water, and shake well) as the dissolution medium; use the paddle method at a speed of 50 revolutions per minute. After 5, 10, 15, 20, 30, 45, and 60 minutes, take the dissolution solution and filter it through a microporous filter membrane, and the filtrate is used as the test solution.

[0099] 2.2.3 Chromatographic conditions

[0100] The dissolution test method for tablets containing cabozantinib analogs is carried out by high performance liquid chromatography, and the chromatographic conditions are as follows:

[0101] Chromatographic column: octadecylsilane bonded silica gel column, the length of the chromatographic column is 150 mm, the inner diameter is 4.6 mm, and the size of the filler particles is 2.7 μm;

[0102] Mobile phase: 0.01 mol / L potassium dihydrogen phosphate solution (pH adjusted to 6.0 with potassium hydroxide)-acetonitrile (30:70) as mobile phase, isocratic elution;

[0103] Column temperature: 40°C;

[0104] Flow rate: 1.5ml / min;

[0105] Detection wavelength: 252nm;

[0106] Solvent: acetonitrile-water (70:30)

[0107] 2.2.4 Determination

[0108] Accurately measure 10 μl of the reference solution and the test solution in Examples 2.2.1 and 2.2.2, respectively, inject into a high performance liquid chromatograph, record the chromatogram, and calculate the dissolution amount of each cabozantinib analog at each time point by the peak area according to the external standard method.

[0109] Calculation formula

[0110]

[0111] Where C is the dissolution amount of the test sample, %;

[0112] A X is the main peak area of ​​the test solution;

[0113] is the average value of the correction factor f of the reference solution, f = G' R / A R ;

[0114] G' R Weigh the sample amount for the reference substance (pure);

[0115] V R is the dilution multiple of the reference solution;

[0116] A R is the main peak area in the reference solution;

[0117] 900 is the volume of dissolution medium, ml;

[0118] 25 represents the labeled dosage of cabozantinib analog tablets, 25 mg.

[0119] [Example 3] Specificity test

[0120] Blank excipient solution: weigh 26.4 mg of blank excipient according to the prescription ratio (equivalent to the amount of excipient corresponding to 2.8 mg of the main ingredient), place in a 100 ml volumetric flask, add an appropriate amount of dissolution medium, shake to disperse, sonicate for 5 minutes, cool, dilute to the scale with dissolution medium, shake well, and filter.

[0121] The blank dissolution medium and blank auxiliary material solution were tested respectively using the chromatographic conditions summarized in Example 2, and the chromatograms were recorded, which were respectively Figure 3-4 shown.

[0122] Results: No interfering peaks were detected in the blank dissolution medium and blank excipient solution at the corresponding positions of the main peak, indicating that this method has good specificity for the dissolution determination of this product.

[0123] [Example 4] System suitability test

[0124] Take 10 μl of the reference solution in Example 2.2.1 and inject it into a high performance liquid chromatograph. Perform the test according to the chromatographic conditions summarized in Example 2.2.3 and record the chromatogram. The chromatographic test results are shown in Figure 1 And Table 10.

[0125] Table 10 System suitability test results

[0126]

[0127]

[0128] In the chromatogram of the reference solution, the theoretical plate number of the main peak was greater than 2000, and the RSD of the main peak area after 5 consecutive injections was less than 2.0%, meeting the system suitability requirements.

[0129] [Example 5] Filter membrane adsorption test

[0130] Test solution: Take the product and operate according to the dissolution curve determination method. After 30 minutes, take a sample and divide it into two parts. One part uses a microporous filter membrane (PES water system), discards 3ml, 5ml, and 10ml of the initial filtrate, and takes the subsequent filtrate as the test solution; the other part is centrifuged for 5min (10000 rpm), and the supernatant is taken as the test solution.

[0131] At least one injection of blank solvent was performed, and one injection of each of the test solution obtained by centrifugation and filtration was performed, and the chromatogram was recorded and the change in the peak area of ​​the main peak was calculated. The results are shown in Table 11.

[0132] Table 11 Filter membrane adsorption test results

[0133]

[0134] Results: After testing and calculation, the change in the peak area of ​​the main peak under different treatment methods was no more than 2.0%. The filtration should have no interference with the dissolution determination and meet the test requirements.

[0135] [Example 6] Linearity and range

[0136] 6.1 Preparation of linear solution

[0137] Linear stock solution (0.694 mg / ml): Take about 36.0 mg of the cabozantinib analog reference substance, accurately weigh it, place it in a 50 ml volumetric flask, add an appropriate amount of solvent to dissolve and dilute to the scale, and shake well.

[0138] Linear series solutions: Accurately measure an appropriate amount of linear stock solution or linear solution, and dilute with dissolution medium according to Table 12 to make solutions containing approximately 2.78 μg, 6.94 μg, 13.89 μg, 27.78 μg, and 41.67 μg of cabozantinib analogs per 1 ml (equivalent to 10%, 25%, 50%, 100%, and 150% of the concentration of the test solution, respectively).

[0139] Table 12 Linear solution preparation list

[0140]

[0141]

[0142] 6.2 Determination

[0143] Accurately measure 10 μl of each of the above linear series solutions, inject them into the liquid chromatograph, record the chromatogram, plot with the peak area (A) as the ordinate and the main component concentration (C) as the abscissa, and calculate the regression equation and correlation coefficient. Figure 6 And Table 13.

[0144] Table 13 Linearity test results

[0145]

[0146] Results: The linear equation correlation coefficient R of cabozantinib analogs was greater than 0.990, the intercept was less than 25%, and the residual standard deviation was less than 10%, indicating that cabozantinib analogs had a good linear relationship in the concentration range of 2.8066 μg / ml to 42.0991 μg / ml (equivalent to 10.0986% to 151.4792% of the test substance).

[0147] [Example 7] Precision test

[0148] The reference solution and the 30-min test solution were prepared according to the preparation method of Example 2.2, 10 μl of the above solution was accurately measured and injected into the liquid chromatograph, and the solubility of the cabozantinib analog was calculated as the repeatability test result. The results are shown in Table 14.

[0149] Table 14 Repeatability test results

[0150] Repeatability 1 2 3 4 5 6 RSD% Dissolution (%) 94.11 80.72 93.44 88.49 95.67 86.45 6.4

[0151] Results: In the 6 test solutions, the RSD of the measured solubility of cabozantinib analogs was less than 10.0%, indicating that the method had good repeatability.

[0152] The product was tested by different people, different equipment and different dates according to the repeatability method, and the RSD of the dissolution of cabozantinib analogs was calculated as the intermediate precision test result. The results of the repeatability and intermediate precision tests were combined to calculate the RSD of the dissolution of the twelve samples. The results are shown in Table 15.

[0153] Table 15 Intermediate precision results

[0154]

[0155] Results: A and B were the operators. The RSD of the dissolution rates of the 12 test solutions was less than 10.0%, indicating that the intermediate precision of this method was good.

[0156] [Example 8] Accuracy test

[0157] 8.1 Solution preparation

[0158] Reference solution Take about 18 mg of cabozantinib analog reference substance, accurately weigh it, place it in a 25 ml volumetric flask, add an appropriate amount of solvent to dissolve and dilute to the mark, shake well, and use it as the reference stock solution. Accurately measure 1 ml of the reference stock solution, place it in a 25 ml volumetric flask, dilute to the mark with the dissolution medium, shake well, and make a solution containing about 27.8 μg per 1 ml as the reference solution.

[0159] 45% test solution: Accurately weigh about 11.7 mg of cabozantinib analog reference substance, add about 106.9 mg of blank excipient mixed powder according to the formulation prescription ratio, add 900 ml of pH 1.0 dissolution medium, measure according to the law, sample after 60 minutes, filter, and take the filtrate as the 45% test solution. Prepare three copies in parallel.

[0160] 75% test solution: Accurately weigh about 19.4 mg of cabozantinib analog reference substance, add about 178.1 mg of blank excipient mixed powder according to the formulation prescription ratio, add 900 ml of pH 1.0 dissolution medium, measure according to the law, sample after 60 minutes, filter, and take the filtrate as the 75% test solution. Prepare three copies in parallel.

[0161] 105% test solution: Accurately weigh about 27.2 mg of cabozantinib analog reference substance, add about 249.4 mg of blank excipient mixed powder according to the formulation prescription ratio, add 900 ml of pH 1.0 dissolution medium, measure according to the law, sample after 60 minutes, filter, and take the filtrate as the 105% test solution. Prepare three copies in parallel.

[0162] 8.2 Determination

[0163] Take 10 μl of the reference solution and each test solution respectively, inject into the liquid chromatograph, and record the chromatogram. Calculate the recovery rate of the main component in each test solution by the peak area according to the external standard method. The results are shown in Table 16.

[0164] Calculation formula:

[0165]

[0166] In the formula

[0167] A X is the main peak area of ​​the test solution;

[0168] is the average value of the correction factor f of the reference solution, f = G' R / A R ;

[0169] G' R Weigh the sample amount for the reference substance (pure);

[0170] VR is the dilution multiple of the reference solution;

[0171] 900 is the volume of dissolution medium, ml;

[0172] A R is the main peak area in the reference solution.

[0173] Table 16 Accuracy results

[0174]

[0175] Results: The recovery rate of the main component was 95.8%, within the range of 95% to 105%, and the RSD value of the recovery rate was 0.4%, which was less than 2.5%, indicating that the method had good accuracy.

[0176] [Example 9] Solution stability

[0177] Accurately measure 10 μl of the reference solution and the test solution in Example 2.2, and inject them into the liquid chromatograph at 0h, 2h, 4h, 6h, 8h, 10h, 12h, 16h, 20h, and 24h, respectively. Record the chromatogram and calculate the change in the peak area of ​​the main peak. The results are shown in Table 17.

[0178] Table 17 Solution stability results

[0179]

[0180]

[0181] Compared with the stability solution at 0h, the changes in the main peak area in the reference solution and the test solution within 24h were all less than 2.0%, and the solution had good stability and met the detection requirements.

[0182] [Example 10] Durability test

[0183] The durability test was carried out on different types of chromatographic columns with column temperature ±2°C, flow rate ±0.1ml / min, mobile phase pH ±0.5.

[0184] The reference solution was prepared according to the preparation method of Example 2, and 10 μl was accurately measured and injected into the liquid chromatograph. The test was performed after appropriate adjustments based on the chromatographic conditions summarized in Example 2.2.3. The test results are shown in Table 18.

[0185] Table 18 Durability test results

[0186]

[0187] The results showed that when all parameters changed slightly, the theoretical plate number of the main peak and the RSD of the main peak area of ​​5 consecutive injections met the system suitability requirements.

Claims

1. A method for determining the dissolution rate of a solid preparation of a cabozantinib analog, characterized in that: The method comprises the following steps: using pH 1.0 hydrochloric acid as a dissolution medium, adopting a paddle method, with a rotation speed of 40 to 60 revolutions per minute, filtering the dissolution solution at different time points, taking the subsequent filtrate as a test solution, and then determining the content of the cabozantinib analog in the test solution, and then comparing it with the labeled amount of the cabozantinib analog in the solid preparation of the cabozantinib analog to calculate its solubility.

2. The method according to claim 1, characterized in that The rotation speed is 50 revolutions per minute.

3. The method according to claim 1, characterized in that The times for taking the dissolution solution are 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes and 60 minutes respectively.

4. The method according to claim 1, characterized in that The cabozantinib analog solid preparation is a cabozantinib analog tablet.

5. The method according to claim 1, characterized in that The steps of the method are: (1) Prepare the dissolution medium of hydrochloric acid at pH 1.0: Take 9 ml of concentrated hydrochloric acid, dilute to 1000 ml with water, and shake well; (2) preparing a test solution: using the dissolution medium obtained in step (1) to dissolve the cabozantinib analog solid preparation, using a slurry method, wherein the rotation speed is 40-60 revolutions per minute, taking the dissolution solution at 5, 10, 15, 20, 30, 45, and 60 minutes, respectively, filtering through a microporous filter membrane, and taking each subsequent filtrate as the test solution; (3) Preparation of reference solution: accurately weigh an appropriate amount of cabozantinib analog reference substance and place it in a volumetric bottle, add acetonitrile-water (70:30) to dissolve and dilute to the scale, shake well, accurately measure an appropriate amount, and then dilute with the dissolution medium to make a solution containing approximately 27.8 μg of cabozantinib analog per 1 ml, shake well, and use this as the reference solution; (4) Determine the dissolution rate: Use high performance liquid chromatography to detect the reference solution and each of the test solution; calculate the dissolution rate at different time points by peak area according to the external standard method.

6. The method according to claim 1, characterized in that The content of cabozantinib analogs in the test solution was determined by high performance liquid chromatography.

7. The method according to claim 5 or 6, characterized in that: The high performance liquid chromatography conditions are as follows: the chromatographic column is an octadecylsilane bonded silica gel column, the mobile phase is a potassium dihydrogen phosphate solution-acetonitrile with a volume ratio of 30:70 and a pH value of 6.0, isocratic elution, the concentration of the potassium dihydrogen phosphate solution is 0.01 mol / L, the column temperature of the chromatographic column is 40°C, the flow rate is 1.5 ml / min, and the detection wavelength is 252 nm.

8. The method according to claim 7, characterized in that The chromatographic column used in the high performance liquid chromatography method was Agilent Poroshell 120BonusRP.

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