Method for determining in-vitro dissolution behavior of fenerenone tablet by flow cell method

Abstract

The invention discloses a determination method of a dissolution curve of a fenerenone tablet, a similarity evaluation method and application thereof, the dissolution rates of the fenerenone tablet at different time points are determined by adopting a closed-loop or open-loop system of a flow cell under different conditions, the dissolution curve is drawn, and then the similarity of the dissolution curve is evaluated through a similarity factor f2. According to the invention, a loading mode of the fenerenone tablet is constructed, so that the fenerenone tablet can be uniformly released, efficient dissolution can be favorably maintained, and the dissolution behavior of a medicine in a body can be better simulated. The dissolution curve of the fenerenone tablet measured through the flow cell method is high in distinguishing capacity, the difference between the imitated preparation and the reference preparation can be effectively distinguished, and consistency evaluation on the imitated preparation and the reference preparation is achieved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for determining the in-vitro dissolution behavior of finerenone tablets by using a flow cell method. BACKGROUND

[0002] After oral administration of solid preparations, the absorption of drugs depends on the dissolution or release of drugs from the preparation, the dissolution of drugs under physiological conditions, and the permeation in the gastrointestinal tract. Since the dissolution and dissolution of drugs have important influence on absorption, in-vitro dissolution test can predict its in-vivo behavior. Similar dissolution curves cannot completely prove the same bioequivalence, but can greatly improve the success rate of bioequivalence test (BE test), and can reduce the risk of clinical efficacy difference between the two.

[0003] Finerenone, molecular formula: C 21 H 22 N4O3, molecular weight: 378.43, chemical name: (4S)-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-formamide. Finerenone is a non-steroidal selective mineralocorticoid receptor (MR) antagonist, which is used for adult patients with chronic kidney disease related to type 2 diabetes (estimated glomerular filtration rate [eGFR] ≥ 25 to < 75 mL / min / 1.73 m 2 2 with albuminuria), which can reduce the risk of continuous decline in eGFR and end-stage renal disease.

[0004] Finerenone tablets are a biological pharmaceutical classification (BCS) II, which is characterized by low solubility-high permeability. After oral administration, finerenone is completely absorbed, and after metabolism, the absolute bioavailability is 44%, and the absorption is not affected by food. However, since finerenone tablets are released quickly in the body, the traditional paddle method cannot simulate the release process of the drug in the in-vivo environment due to the single mechanical movement form, and the problems such as floating, stacking and adhesion of the sample during the release process. The composition and pH of the medium do not change with time during the experiment, which is quite different from the actual situation in the body, and cannot better simulate the release of immediate-release preparations in the body.

[0005] In the case of consistency of the dissolution curve of the traditional dissolution determination method (paddle method) and the reference preparation, the domestic finerenone tablets currently reported have lower success rate through clinical bioequivalence test. Therefore, in order to make a prediction for in-vivo bioequivalence test and improve the success rate of clinical bioequivalence test, it is important to establish a strong in-vitro dissolution detection method for the consistency evaluation of finerenone tablets. SUMMARY

[0006] The present application aims to provide a method for determining the dissolution curve of finerenone tablets, and to evaluate the quality of the generic preparation by comparing the similarity of the dissolution curves of the generic preparation and the reference preparation in vitro. The method can accurately and efficiently obtain the dissolution characteristics of finerenone tablets, which are close to the dissolution mode in the human body, greatly increasing the reliability of the in vitro method in predicting in vivo behavior, and effectively guiding the success of the BE experiment.

[0007] To achieve the above-mentioned object, the present application adopts the following scheme:

[0008] The present application provides a method for determining the in vitro dissolution of finerenone tablets, which uses a flow cell method to determine the dissolution of finerenone tablets, comprising the following steps:

[0009] The dissolution medium is pumped into the flow cell to release the finerenone tablets in the flow cell, and the dissolution solution is output to the sampling device. The samples in the sampling device are collected at a set time;

[0010] The flow cell is a closed loop flow cell or an open loop flow cell;

[0011] When the flow cell is a closed loop flow cell, the step of releasing the finerenone tablets is:

[0012] Place 5mm glass beads at the bottom of the flow cell, and 1mm glass beads on top. Place the finerenone tablets on the glass beads, and 1mm glass beads on top. Install a filter membrane on the top of the flow cell and seal it. The dissolution medium flows from bottom to top through the constant temperature flow cell body. The volume of the dissolution medium is 900mL, the flow rate is 8mL / min, the temperature is 37.0±0.5℃, and the concentration of finerenone in the dissolution solution is determined at a fixed time point until the dissolution of the finerenone tablets is complete;

[0013] When the flow cell is an open loop flow cell, the step of releasing the finerenone tablets is:

[0014] Place 5mm glass beads at the bottom of the flow cell, and 1mm glass beads on top. Place the finerenone tablets on the glass beads, and 1mm glass beads on top. The dissolution medium passes through the flow cell, and the dissolution medium is sequentially switched. The flow rate is 4-16mL / min, the temperature is 37.0±0.5℃, and the concentration of finerenone in the dissolution solution is determined at a fixed time point until the dissolution of the finerenone tablets is complete;

[0015] The dissolution of finerenone in the dissolution solution at different time points is determined by high performance liquid chromatography, and the dissolution curve of the cumulative dissolution amount versus time is drawn.

[0016] Further, when the high performance liquid chromatography is used to determine the dissolution amount of finerenone in the dissolution liquid, the parameters of the liquid chromatograph are as follows: mobile phase: 10 mmol / L potassium dihydrogen phosphate (pH value is adjusted to 3.0 by phosphoric acid)-acetonitrile (70:30); chromatographic column: Intersil ODS-3 (4.6 mm*50 mm, 5 μm); flow rate: 1.0 ml / min; column temperature: 30 DEG C; detection wavelength: 250 nm; injection volume: 10 μL.

[0017] Further, when the flow cell is a closed loop flow cell, the medium is pH 1.2 hydrochloric acid solution, pH 4.5 acetate buffer solution, pH 6.8 phosphate buffer solution and water.

[0018] Further, when the flow cell is a closed loop flow cell, the sampling time points are 10 min, 20 min, 30 min, 45 min, 60 min, 120 min, 180 min and 240 min.

[0019] Further, when the flow cell is an open loop flow cell, the sequentially switched dissolution medium is pH 1.2 hydrochloric acid solution (30 min)-pH 4.5 acetate buffer solution (90 min)-pH 6.8 phosphate buffer solution (60 min)-water (60 min).

[0020] Further, when the flow cell is an open loop flow cell, the sampling time points are 10 min, 20 min, 30 min, 45 min, 60 min, 120 min, 180 min and 240 min.

[0021] The similarity evaluation method of the dissolution curve of the finerenone tablet is as follows: the similarity factor method is used to calculate the similarity factor f2 by using the average dissolution data of the finerenone generic preparation and the reference preparation at different time points, and the similarity factor f2 is used to evaluate the similarity of the dissolution curve of the finerenone generic preparation and the reference preparation; when the similarity factor f2 is greater than or equal to 50, the dissolution curves of the generic preparation and the reference preparation are similar, and when the similarity factor f2 is less than 50, the dissolution curves of the generic preparation and the reference preparation are not similar. The calculation formula of the similarity factor is as follows:

[0022]

[0023] Wherein, Rt is the average cumulative release of the reference preparation at the t time point; Tt is the average cumulative release of the test preparation at the t time point; n is the number of sampling time points.

[0024] The application further discloses application of the method for determining the in-vitro dissolution behavior of the finerenone tablets or the similarity evaluation method of the dissolution curve of the finerenone tablets to consistency evaluation of reference preparations and generic preparations of the finerenone tablets.

[0025] In the embodiments of the application, the consistency of the in-vitro dissolution behavior of the reference preparations and the generic preparations of the finerenone tablets is investigated by three different dissolution methods, namely the basket paddle method, the flow-through cell closed loop system and the open loop system. The dissolution curves of the reference preparations and the generic preparations measured by the basket paddle method are similar, which is one of the reasons why many pharmaceutical manufacturers have lower clinical bioequivalence test results. However, the dissolution curves of the reference preparations and the generic preparations measured by the flow-through cell method have a certain degree of distinction, which can indicate that the in-vitro dissolution behavior of the generic preparations and the reference preparations is not similar, and can effectively evaluate the quality of the generic preparations and realize consistency evaluation of the generic preparations and the reference preparations. In addition, the dissolution conditions of the method are closer to the in-vivo physiological environment, and therefore the method can be used to establish the correlation between the in-vitro dissolution curve of a drug and the in-vivo pharmacokinetics, can predict the absorption and bioavailability of the drug in the body, and thus reduce the number of animal experiments and clinical tests and reduce the research and development cost.

[0026] Compared with the prior art, the application has the following beneficial effects:

[0027] The application develops a new dissolution determination method of the finerenone tablets, utilizes the open loop and the closed loop systems of the flow-through cell to determine the dissolution curve, and utilizes the similarity factor method to distinguish the dissolution behavior of the reference preparations and the generic preparations. The application constructs a loading mode of the finerenone tablets, i.e. 7g of 1mm glass beads are laid on the 5mm glass beads, the finerenone tablets are placed on the glass beads, 2.5g of 1mm glass beads are laid on the upper layer of the glass beads, the fluid can continuously flow in parallel without mutual interference, the surface shear force is smaller, the dissolution is relatively slower, the finerenone tablets are uniformly released, and the high-efficiency dissolution can be maintained, which is favorable for better simulating the dissolution behavior of the drug in the body. The dissolution speed can be changed by changing the flow rate of the dissolution medium, the difference in experimental phenomena can be better observed, the prescription or process difference can be effectively distinguished, the amount of the dissolution medium used in the open loop system is unlimited, a low-concentration surfactant does not need to be used, the leakage tank condition can be maintained all the time by increasing the amount of the dissolution medium, and the pH change of the gastrointestinal tract is simulated, and therefore the method is especially suitable for in-vitro dissolution and in-vitro and in-vivo correlation research of poorly soluble drugs. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 For the specificity test, the HPLC atlas is as follows: A is a blank excipient solution, B is a test sample solution, and C is a control sample solution.

[0029] Figure 2 is the dissolution curve in different dissolution media in the paddle method: a is pH 1.2 hydrochloric acid solution, b is pH 4.5 acetate buffer solution, c is pH 6.8 phosphate buffer solution, and d is water.

[0030] Figure 3 The construction of different glass beads from left to right is: mode one, mode two, mode three, mode four, and mode five.

[0031] Figure 4 A dissolution curve of A tablet measured by the flow-through cell method under the construction of different glass beads

[0032] Figure 5 is the dissolution curve in different dissolution media in a closed loop system: a is pH 1.2 hydrochloric acid solution, b is pH 4.5 acetate buffer solution, c is pH 6.8 phosphate buffer solution, and d is water.

[0033] Figure 6 The dissolution curve of the reference sample and the generic preparation at a flow rate of 4 mL / min in an open loop system.

[0034] Figure 7 The dissolution curve of the reference sample and the generic preparation at a flow rate of 8 mL / min in an open loop system.

[0035] Figure 8 The dissolution curve of the reference sample and the generic preparation at a flow rate of 16 mL / min in an open loop system. DETAILED DESCRIPTION

[0036] The present application will be further described in detail below in conjunction with test examples and specific embodiments. However, it should not be understood that the protection scope of the present application is limited to the following examples, and any technology realized based on the present application falls within the scope of the present application.

[0037] The reference preparation used in the present application is Finerenone tablet (10 mg, available from ® Kerendia ® ) produced by Bayer AG); the generic preparation is 3 different domestic nonnalineone generic preparations (10 mg), named generic preparation A, generic preparation B, and generic preparation C.

[0038] Example 1: Determination of the dissolution curve of nonnalineone tablet by paddle method

[0039] 1. Establishment of nonnalineone method

[0040] Chromatographic conditions: mobile phase: 10 mmol / L potassium dihydrogen phosphate (pH value is adjusted to 3.0 with phosphoric acid)-acetonitrile (70:30); chromatographic column: Intersil ODS-3 (4.6 mm x 50 mm, 5 μm) or a chromatographic column with equivalent performance; flow rate: 1.0 ml / min; column temperature: 30°C; detection wavelength: 250 nm; sample injection volume: 10 μl;

[0041] Preparation of the reference solution: about 22 mg of finerenone reference was accurately weighed into a 20-ml flask, dissolved and diluted to the mark with acetonitrile by ultrasonic, shaken well, and 2 ml was accurately taken into a 200-ml flask, diluted to the mark with the dissolution medium, shaken well (two preparations were made in parallel).

[0042] Preparation of the blank excipient solution: the other excipients except for the finerenone prescription amount were accurately weighed according to the preparation method of the reference, placed in a 20-ml flask, dissolved and diluted to the mark with acetonitrile by ultrasonic, shaken well, and 2 ml was accurately taken into a 200-ml flask, diluted to the mark with the dissolution medium, shaken well, and the blank excipient solution was obtained.

[0043] Preparation of the test solution: 20 finerenone tablets were finely ground, accurately weighed (equivalent to 10 mg of finerenone), placed in a 20-ml flask, dissolved with acetonitrile by ultrasonic, diluted to the mark with water, shaken well, accurately took 2 ml into a 200-ml flask, diluted to the mark with the dissolution medium, shaken well, and the test solution was obtained.

[0044] Specificity test: an appropriate amount of the reference solution, test solution and blank excipient solution were taken, filtered with a 0.22-μm organic filter membrane, and determined by HPLC sample injection to obtain the corresponding chromatograms, as shown in Figure 1 From the figure, it can be seen that the peaks of the blank excipient and the solvent can be well separated from the baseline of the main drug. The method has good specificity.

[0045] 2-paddle method for determining the dissolution curve of finerenone tablets

[0046] Dissolution curve test solution: 6 tablets were taken and placed in 6 dissolution cups, respectively, according to the dissolution and release determination method (Chinese Pharmacopoeia 2020 edition, four general rules 0931, second method), 900 ml of pH 1.2 hydrochloric acid solution, pH 4.5 acetic acid solution and pH 6.8 phosphate solution were used as dissolution medium, the rotation speed was 50 revolutions per minute, and the medium temperature was 37°C ± 0.5°C. According to the law, sample was taken at the specified time, 10 ml of dissolution liquid was taken, and the same volume of dissolution medium at the same temperature was supplemented, filtered (polyether sulfone, 0.45 μm), 8 ml of the initial filtrate was discarded, and the subsequent filtrate was taken as the test solution.

[0047] Sampling time of pH 1.2 hydrochloric acid solution: 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 60 min, 90 min, 120 min.

[0048] Sampling time of pH 4.5 acetic acid solution: 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 60 min.

[0049] Sampling time of pH 6.8 phosphate buffer solution: 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 60 min, 90 min.

[0050] Sampling time of water medium: 5 min, 10 min, 15 min, 20 min, 30 min, 45 min.

[0051] The liquid chromatograph was injected according to the above-mentioned chromatographic conditions, and the chromatogram was recorded. The dissolution was calculated by peak area according to the external standard method. The dissolution curves of each medium are shown in Figure 2.

[0052] The similarity factor calculation formula is:

[0053]

[0054] Wherein, Rt is the average cumulative release of the reference preparation at the t time point; Tt is the average cumulative release of the test preparation at the t time point; n is the number of sampling time points.

[0055] The similarity factor (f2) is used to evaluate the similarity of three generic preparations and the reference preparation. When 50 ≤ f2≤ 100, it is considered that the release curves of the reference preparation and the generic preparation are similar. The time points of pH 1.2 hydrochloric acid solution are 30 min, 45 min, 60 min, 90 min; the time points of pH 6.8 acetic acid buffer are 20 min, 30 min, 45 min, 60 min; the time points of water are 10 min, 15 min, 20 min, 30 min. Using the average dissolution data of the generic preparation and the reference preparation, the value of similarity factor f2 is calculated,

[0056] The similarity results are shown in Tables 1-4.

[0057] Table 1 Calculation results of pH 1.2 hydrochloric acid solution dissolution medium

[0058] Table 2 Calculation results of pH 4.5 acetic acid buffer dissolution medium

[0059]

[0060] Table 3 pH 6.8 Phosphate Buffer Dissolution Medium Calculation Results

[0061]

[0062] Table 4 Aqueous Dissolution Medium Calculation Results

[0063]

[0064] From Tables 1-4, according to the Technical Guidelines for Dissolution Test of Ordinary Oral Solid Preparations, in pH 4.5 acetate buffer, the average dissolution of the test sample and the reference sample at 15 minutes was not less than 85%, and the dissolution curves were considered similar. In the other three dissolution media, the similarity factor f2 of the three generic preparations and the reference preparation was greater than 50, indicating that the dissolution curves of the generic preparations and the reference preparation were relatively similar. Using this method for consistency evaluation of non-neliriketone tablets, the discrimination was poor, and the differences between the generic preparations and the reference preparation could not be effectively evaluated.

[0065] Example 2 Determination of the dissolution curve of non-neliriketone tablets by a flow cell closed system

[0066] 1. Consideration before the experiment: selection of flow cell system: open loop, closed loop; selection of flow cell and sample loading method; flow rate of dissolution medium; filtration method; split ratio needs to be considered for open loop.

[0067] 2. Selection of flow cell and sample loading method: the inner diameter of the standard flow cell is generally 12mm / 6ml and 22.6mm / 14ml, and different pool bodies are selected for different dosage forms. It was found that for the same non-neliriketone tablet sample, using two different diameter standard flow cells would produce a difference of more than three times the average dissolution rate, so for non-neliriketone tablets, the flow rate was increased and the standard small pool was used to improve the dissolution rate.

[0068] 3. Glass bead loading method: under the closed loop system, taking manufacturer A tablets as an example, comparing five different glass bead loading methods (such as Figure 3 ), the dissolution curves of the above five loading methods in pH 4.5 acetate dissolution medium were drawn. The dissolution curves are shown in Figure 4Method 1: Place A tablet directly on 5 mm glass beads soaked with water; Method 2: Place 1 mm glass beads on top of 5 mm glass beads to the section of the pool body and place A tablet on the glass beads; Method 3: Place 1 mm glass beads on top of 5 mm glass beads to the section of the pool body, and place the tablet upside down on the tablet pressing clamp. Method 4: Place 7 g of 1 mm glass beads on top of 5 mm glass beads, place A tablet on the glass beads, and place 2.5 g of 1 mm glass beads on top. Method 5: Place 2.5 g of 1 mm glass beads on top of 5 mm glass beads, place A tablet on the glass beads, and place 7 g of 1 mm glass beads on top.

[0069] Through comparative experiments, Method 1 is a pool body without adding glass beads, which is a turbulent flow mode, a fast and multi-directional irregular flow, and the dissolution is too fast. Methods 2, 3, 4, and 5 are pool bodies with added glass beads, and the fluid continuously flows in parallel without mutual interference, which is a laminar flow mode with smaller surface shear force, and the dissolution is relatively slow. Through comparative experiments, Methods 2 and 3 release slowly and unevenly. Methods 4 and 5 can make tablet A release uniformly, and Method 5 releases fast in the early stage, but the dissolution rate is inhibited in the middle and late stages due to the inhibition of medium dissolution. Method 4 can help maintain high dissolution efficiency. Therefore, the dissolution experiment is carried out by using the transfer method of Method 4.

[0070] 4. Filtration method: The test solution is filtered in real time when flowing out of the flow-through pool. A filter membrane with appropriate pore size and material is selected to prevent excessive negative pressure, but also to trap undissolved particles. Common filter membrane materials include glass fiber, cellulose, polypropylene, etc. Through multiple experiments, a glass fiber filter membrane (0.45-0.7-2.7 μm) is installed at the top of the pool and sealed. After the water bath, the dissolution medium flows from bottom to top through the pool body, dissolves the sample, and flows out after filtration. The eluate is collected at regular intervals.

[0071] 5. Determine the dissolution curve of finerenone tablets using a flow-through pool closed system: 900 mL of four different dissolution media is treated by a degassing machine and placed in the solvent reservoir. The stirring speed in the solvent area is 600 revolutions, and the temperature is 37 ± 0.5°C. The flow rate is 8 ml / min, and the dissolution medium flows through the pool body through a constant temperature water bath. The four media are pH 1.2 hydrochloric acid solution, pH 4.5 acetic acid salt buffer, pH 6.8 phosphate buffer, and water medium.

[0072] 6. The sampling points are: 10 min, 20 min, 30 min, 45 min, 60 min, 120 min, 180 min, and 240 min. The sampling volume is 10 mL, and the supplement is 10 mL.

[0073] 7. Determine the dissolution at the above sampling points by high performance liquid chromatography, and the chromatographic conditions are the same as in Example 1. The dissolution curves in different media are plotted, as shown in Figure 5.

[0074] As shown in Figure 5, the dissolution curves of the three generic formulations and the reference formulation were measured by the closed loop system with flow-through cell method. The release behaviors of the three generic formulations were quite different from the reference formulation. Compared with the paddle method, it was found that the dissolution rate of the paddle method was faster than that of the flow-through cell method in the same dissolution medium. This faster dissolution curve may be because the paddle method provides faster stirring and mixing than the flow-through cell method. The great difference in in vitro dissolution behavior will certainly lead to a great difference in in vivo absorption behavior.

[0075] The similarity factor method was used to compare the similarity of the dissolution curves of the three non-neliriketone generic formulations and the reference formulation. The pH 1.2 hydrochloric acid solution selected time points 30 min, 45 min, 60 min, 120 min, 180 min; pH 4.5 acetate buffer solution selected time points 45 min, 60 min, 120 min, 180 min; pH 6.8 acetate buffer solution selected time points 30 min, 45 min, 60 min, 120 min; water selected time points 45 min, 60 min, 120 min, 180 min, using the average dissolution data of the generic formulation and the reference formulation, the value of the similarity factor f2 was calculated, the results were shown in Tables 5-8.

[0076] Table 5 Closed loop system pH 1.2 hydrochloric acid solution dissolution medium calculation results

[0077]

[0078] Table 6 Closed loop system pH 4.5 acetate buffer solution dissolution medium calculation results

[0079]

[0080] Table 7 Closed loop system pH 6.8 phosphate buffer solution dissolution medium calculation results

[0081]

[0082] Table 8 Closed loop system water dissolution medium calculation results

[0083]

[0084] As shown in Tables 5-8, the similarity factor f2 of the three generic formulations and the reference formulation in four dissolution media measured by the closed loop system with flow-through cell method was only greater than 50 when the dissolution medium was pH 4.5 acetate buffer solution, indicating that the dissolution curves of the generic formulations and the reference formulation were not similar. It showed that the flow-through cell method had better discrimination than the conventional paddle method.

[0085] Example 3 Determination of the dissolution curve of non-neliriketone tablets by flow-through cell open loop system

[0086] 1. The sample to be tested is tested in the same way as the closed loop system, and the amount of dissolution medium in the open system of the flow cell method is not limited, and the dissolution medium is always fresh during the process, and the dissolution part will be taken away from the flow cell in time. By increasing the amount of dissolution medium, the leakage groove condition can be maintained. The pH value is automatically changed by sequence inertia from pH 1.2 to pH 4.5 to pH 6.8 to water.

[0087] 2. The flow rate is set to 4, 8, 16 ml / min. The total volume of the dissolution medium required is calculated according to the sampling time and the flow rate, and the sampling volume of each sampling point is calculated according to the shunt ratio. The pH value is automatically changed during the experiment, and the dissolution medium switched in time sequence is pH 1.2 hydrochloric acid solution (30 min)-pH 4.5 acetic acid buffer (90 min)-pH 6.8 phosphoric acid buffer (60 min)-water (60 min).

[0088] 3. The above flow cell is an open loop, and the sampling points are set to: 10 min, 20 min, 30 min, 45 min, 60 min, 120 min, 180 min, 240 min. The sampling volumes are 40 mL, 40 mL, 40 mL, 60 mL, 60 mL, 240 mL, 240 mL, and 240 mL, respectively.

[0089] 4. The dissolution of the above sampling points is determined by high performance liquid chromatography, and the dissolution curves in different media are drawn. The chromatographic conditions are the same as in Example 1, and the results are shown in Figures 6-8 .

[0090] From Figures 6-8 it can be seen that under the open loop system of the flow cell, the change of the flow rate has a certain influence on the dissolution rate and the cumulative dissolution amount of the reference preparation. Under three different flow rates, the release curves of the three generic preparations are significantly different from the release curve of the reference preparation, and the difference is most obvious when the flow rate is 8 ml / min; the above results show that the dissolution curve of the flow cell open loop system for determining the dissolution curve of the non-neliriketone tablet can effectively distinguish the difference between the generic preparation and the reference preparation.

[0091] The similarity factor f2 is calculated by using the similarity factor method, which is used to compare the similarity of the dissolution curves of the generic preparation of non-neliriketone tablets and the reference preparation, wherein the flow rate is 4 mL / min, and the time points are 45 min, 60 min, 120 min, and 180 min; the flow rate is 8 mL / min, and the time points are 30 min, 45 min, 60 min, and 120 min; the flow rate is 16 mL / min, and the time points are 30 min, 45 min, 60 min, and 120 min; the average dissolution data of the generic preparation and the reference preparation are used to calculate the value of the similarity factor f2; the results are shown in Tables 9-11.

[0092] Table 9. Nonalineone tablet dissolution test results at 4 mL / min flow rate for open system

[0093]

[0094] Table 10. Nonalineone tablet dissolution test results at 8 mL / min flow rate for open system

[0095]

[0096] Table 11. Nonalineone tablet dissolution test results at 16 mL / min flow rate for open system

[0097]

[0098] From Tables 9-11, the dissolution curves of the three generic preparations and the reference preparation were measured by the open system with flow-through cell method in four media, and the similarity factor f2 of the release behavior of the three generic preparations and the reference preparation was less than 50. Compared with the paddle method, the dissolution test was performed by the method of the present embodiment, and the similarity of the three generic preparations A, B and C and the reference preparation was reduced, that is, the method has more distinguishing power.

[0099] In summary, the consistency of the in vitro dissolution behavior of the nonalineone tablet generic preparation and the reference preparation was investigated by three different dissolution methods of the paddle method, the flow-through cell closed system and the open system, respectively, and different test results were obtained. The new flow-through cell dissolution method showed that the in vitro dissolution behavior of the generic preparation and the reference preparation was not similar, while the traditional method showed similarity. The new dissolution method, especially the loading dissolution method of the nonalineone tablet in the flow-through cell, is more similar to the in vivo behavior, has more distinguishing power for the subtle differences between drug preparations, can be used to establish the correlation between the in vitro dissolution curve and the in vivo pharmacokinetics, can predict the absorption and bioavailability of the drug in the body, thereby reducing the number of animal experiments and clinical trials, and reducing the research and development cost.

Claims

1. A method for determining the in vitro dissolution behavior of fenelazol tablets using a flow-through cell method, characterized in that, Includes the following steps: The dissolution medium is pumped into the flow cell to dissolve and release the fenelazol tablets in the flow cell. The dissolution solution is output to the sampling device, and the sample in the sampling device is collected within a set time. The flow cell can be a closed-loop flow cell or an open-loop flow cell. When a closed-loop flow cell is used, the dissolution and release steps for fenelazol tablets are as follows: Place 5mm glass beads at the bottom of the flow-through cell, then lay 1mm glass beads on top, place fenelazol tablets on the glass beads, and lay 1mm glass beads on top of that. Install a filter membrane on the top of the cell and seal it. Use different dissolution media to flow from bottom to top through the constant temperature flow-through cell. The volume of the dissolution media is 900mL, the flow rate is 8mL / min, and the temperature is 37.0±0.5℃. Measure the concentration of fenelazol in the dissolution solution at fixed time points until the fenelazol tablets are completely dissolved. When an open-loop flow cell is used, the dissolution and release steps for fenelazol tablets are as follows: Place 5mm glass beads at the bottom of the flow cell, then lay 1mm glass beads on top, place fenelazol tablets on the glass beads, and lay another 1mm glass beads on top. Install a filter membrane on the top of the cell and seal it. The dissolution medium is passed through the flow cell using a sequentially switching dissolution medium. The flow rate is 4-16 mL / min, and the temperature is 37.0±0.5℃. Measure the concentration of fenelazol in the dissolution solution at fixed time points until the fenelazol tablets are completely dissolved. The dissolution rate of fenelazol in the dissolution solution at different time points was determined by high performance liquid chromatography, and a dissolution curve of cumulative dissolution amount versus time was plotted.

2. The method for determining the in vitro dissolution behavior of fenelazol tablets using a flow-through cell method according to claim 1, characterized in that, When determining the amount of fenelazol dissolved in the solution using high performance liquid chromatography (HPLC), the parameters of the HPLC instrument are as follows: mobile phase: 10 mmol / L potassium dihydrogen phosphate adjusted to pH 3.0 with phosphoric acid: acetonitrile = 70:30; column: Intersil ODS-3, 4.6 mm × 50 mm, 5 μm; flow rate: 1.0 ml / min; column temperature: 30 ℃; detection wavelength: 250 nm; injection volume: 10 µL.

3. The method for determining the in vitro dissolution behavior of fenelazol tablets using a flow-through cell method according to claim 1, characterized in that, When the flow cell is a closed-loop flow cell, the medium is a pH 1.2 hydrochloric acid solution, a pH 4.5 acetate buffer solution, a pH 6.8 phosphate buffer solution, and water.

4. The method for determining the in vitro dissolution behavior of fenelazol tablets using a flow-through cell method according to claim 3, characterized in that, When the flow cell is a closed-loop flow cell, the sampling time points are 10 min, 20 min, 30 min, 45 min, 60 min, 120 min, 180 min, and 240 min.

5. The method for determining the in vitro dissolution behavior of fenelazol tablets using a flow-through cell method according to claim 1, characterized in that, When the flow cell is an open-loop flow cell, the sequentially switched dissolution media are: pH 1.2 hydrochloric acid solution for 30 min - pH 4.5 acetate buffer solution for 90 min - pH 6.8 phosphate buffer solution for 60 min - water for 60 min.

6. The method for determining the in vitro dissolution behavior of fenelazol tablets using a flow-through cell method according to claim 5, characterized in that, When the flow cell is an open-loop flow cell, the sampling time points are 10 min, 20 min, 30 min, 45 min, 60 min, 120 min, 180 min, and 240 min.

7. A method for evaluating the similarity of dissolution curves of fenelazol tablets, characterized in that, The similarity factor method was used to calculate the similarity factor f2 by using the average dissolution data of the generic and reference formulations of glibenclamide tablets at different time points. The similarity factor f2 was used to evaluate the similarity between the dissolution curves of the generic and reference formulations of glibenclamide tablets. When the similarity factor f2 ≥ 50, the dissolution curves of the generic and reference formulations were similar, and when the similarity factor f2 < 50, the dissolution curves of the generic and reference formulations were not similar.

8. The application of the method for determining the in vitro dissolution behavior of fenelitonee tablets by the flow cell method according to any one of claims 1-6 in the consistency evaluation of the reference and generic formulations of fenelitonee tablets.

9. The application of the similarity evaluation method for the dissolution curve of fenelinone tablets as described in claim 7 in the consistency evaluation of the reference formulation and the generic formulation of fenelinone tablets.

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