A model, method and application for determining the penetration degree of a drug to be tested

Abstract

The present disclosure relates to a model, method and application for determining the permeability of a drug to be tested, wherein the model includes a tool drug and a permeability parameter corresponding to the tool drug. The model disclosed herein provides classified drugs in the hyperosmotic interval and corresponding permeability parameters, which can be used to accurately attribute the permeability of unknown compounds, meeting the application in the formulation research and development process; the method for determining the permeability of unknown compounds provided by the present disclosure is simple and easy to implement, and can provide a basis and convenience for the relevant development of formulations.

Description

Technical Field

[0001] The present disclosure relates to the technical field of drug permeability, and in particular to a model, method and application for determining the permeability of a drug to be tested. Background Art

[0002] At present, the research and development of drugs needs to determine their permeability in order to facilitate subsequent development and in vivo investigation. Drug permeability can be divided into high permeability, medium permeability and low permeability according to the degree of in vivo bioavailability. Due to the ethical issues of using human in vivo drug permeability research, it is difficult to carry out drug permeability research in humans. At the same time, due to the species issues between different animals and humans and the long experimental cycle, the application of drugs in animal permeation models is also hindered. Therefore, the use of in vitro permeation models to conduct rapid and effective research on drugs is currently a commonly used method in drug formulation research.

[0003] At present, relevant documents have suggested using permeation classification model drugs to investigate the permeability category of unknown compounds. However, there is no clear tool scale to refer to for determining the degree of drug permeability (high, medium, and low permeability). If inappropriate model drugs are used to calibrate unknown compounds, it is easy to lead to incorrect permeation classification judgments, which will lead to deviations in the understanding of the permeability of unknown compounds, directly affecting subsequent drug development and in vivo drug investigations, and seriously delaying the rate of formulation development.

[0004] At present, metoprolol or propranolol is more commonly used as a classification tool for hyperosmotic drugs. The in vitro permeation results of unknown compounds are compared with the permeation values ​​of the above two drugs. If it is higher than metoprolol or propranolol, it is considered a hyperosmotic drug, and if it is lower, it is considered a hypoosmotic drug. However, the latter is prone to misjudgment, because metoprolol and propranolol are substances with higher permeability in the hyperosmotic range and cannot be used as high- and medium-osmotic classification boundary drugs. Therefore, hyperosmotic drugs are easily misjudged as medium-osmotic drugs, which will directly affect the design and development of subsequent drug preparations, greatly affecting R&D efficiency and even leading to R&D failure.

[0005] Therefore, it would be desirable to provide a method that can determine the extent of penetration of an unknown compound. Summary of the invention

[0006] In order to solve the above technical problems, the present disclosure provides a model, method and application for determining the penetration degree of a drug to be tested. The method can determine the penetration degree of the drug to be tested by using the model.

[0007] In a first aspect, the present disclosure provides a model for determining the permeability of a drug to be tested, wherein the model includes a tool drug and a permeability parameter corresponding to the tool drug.

[0008] The model provided by the present disclosure can be used to determine the permeability of the drug to be tested. When using the model, the permeability parameters of the drug to be tested are compared with the permeability parameters corresponding to the tool drug to determine the size of the permeability of the drug to be tested and the tool drug, and then the permeability degree of the permeability of the drug to be tested is determined.

[0009] In order to determine the penetration degree of the drug to be tested, as a preferred technical solution of the present disclosure, the tool drugs include carbamazepine, propranolol hydrochloride, metoprolol tartrate, phenytoin and minoxidil.

[0010] If the permeability parameter of the drug to be tested is between the permeability parameters of the hypertonic tool drug, the permeability degree of the drug to be tested belongs to hypertonicity (high permeability) and the specific permeability degree of the drug to be tested is confirmed.

[0011] Among the tool drugs provided in the present disclosure, minoxidil can be used as a degree-limited internal standard drug. If the permeability parameter of the drug to be tested is greater than minoxidil, it means that the permeability of the drug to be tested is high permeability.

[0012] As a preferred technical solution of the present disclosure, the permeability parameters corresponding to the tool drug include an effective permeation rate.

[0013] As a preferred technical solution of the present disclosure, the effective permeation rate includes the effective permeation rate corresponding to pH=5.0, the effective permeation rate corresponding to pH=6.5 and the effective permeation rate corresponding to pH=7.4.

[0014] Since the pH value of the human body environment is different, it will have a great impact on the permeability of the drug. Therefore, the present disclosure provides effective permeation rates corresponding to different pH values ​​to confirm the permeation degree of the drug to be tested under different pH values.

[0015] As a preferred technical solution of the present disclosure, the effective permeation rate is determined by a parallel artificial membrane in vitro permeation model.

[0016] In a second aspect, the present disclosure provides application of the model described in the first aspect in determining the penetration degree of a drug to be tested.

[0017] As a preferred technical solution of the present disclosure, the classification of the permeability degree includes high permeability, and preferably the high permeability includes a high permeability high interval, a high permeability middle interval and a high permeability low interval.

[0018] In a third aspect, the present disclosure provides a method for determining the penetration degree of a drug to be tested using the model described in the first aspect, the method comprising:

[0019] (1) Determine the permeability parameters corresponding to the drug to be tested;

[0020] (2) Comparing the permeability parameter corresponding to the drug to be tested with the permeability parameter corresponding to the tool drug included in the model described in the first aspect, thereby determining the permeability degree of the drug to be tested.

[0021] As a preferred technical solution of the present disclosure, the method for determining the penetration degree of the drug to be tested is as follows:

[0022] If the permeability of the drug to be tested is greater than the permeability of minoxidil, the permeability of the drug to be tested is high permeability.

[0023] As a preferred technical solution of the present disclosure, the permeability of the drug to be tested is high permeability, and the method for further determining the permeability of the drug to be tested includes:

[0024] The pH value corresponding to the permeability parameter corresponding to the drug to be tested is determined, and then compared with the permeability parameter corresponding to the same pH value of the tool drug to further determine whether the drug to be tested belongs to the high permeability high interval, the high permeability medium interval or the high permeability low interval.

[0025] As a preferred technical solution of the present disclosure, the pH corresponding to the permeability parameter of the drug to be tested is 5.0, and the method for further determining the permeability of the drug to be tested includes:

[0026] If the permeability of the drug to be tested is greater than the permeability of carbamazepine, the permeability of the drug to be tested is in the high permeability high interval;

[0027] If the permeability of the carbamazepine is greater than the permeability of the drug to be tested and greater than the permeability of the propranolol hydrochloride, the permeability of the drug to be tested is in the high permeability range;

[0028] If the permeability of the propranolol hydrochloride>the permeability of the drug to be tested>the permeability of the minoxidil, the permeability of the drug to be tested is in the high permeability low range.

[0029] As a preferred technical solution of the present disclosure, the pH corresponding to the permeability parameter of the drug to be tested is 6.5, and the method for further determining the permeability of the drug to be tested includes:

[0030] If the permeability of the drug to be tested is greater than the permeability of propranolol hydrochloride, the permeability of the drug to be tested is in the high permeability high range;

[0031] If the permeability of the propranolol hydrochloride>the permeability of the drug to be tested>the permeability of the metoprolol tartrate, then the permeability of the drug to be tested is in the high permeability middle range;

[0032] If the permeability of the metoprolol tartrate>the permeability of the drug to be tested>the permeability of the minoxidil, the permeability of the drug to be tested is in the high permeability low range.

[0033] As a preferred technical solution of the present disclosure, the pH corresponding to the permeability parameter of the drug to be tested is 7.4, and the method for further determining the permeability of the drug to be tested includes:

[0034] If the permeability of the drug to be tested is greater than the permeability of propranolol hydrochloride, the permeability of the drug to be tested is in the high permeability high range;

[0035] If the permeability of the propranolol hydrochloride>the permeability of the drug to be tested>the permeability of phenytoin, then the permeability of the drug to be tested is in the high permeability middle range;

[0036] If the permeability of phenytoin>the permeability of the drug to be tested>the permeability of minoxidil, the permeability of the drug to be tested is in the high permeability low range.

[0037] Compared with the prior art, the technical solution provided by the embodiments of the present disclosure has the following advantages:

[0038] (1) The model disclosed in the present invention provides classification drugs and corresponding permeability parameters in three intervals of hyperosmotic zone, mesoosmotic zone and hypoosmotic zone in the hyperosmotic model drug, which can be used to accurately attribute the permeability of unknown compounds and meet the application in the preparation research and development process;

[0039] (2) The method for determining the penetration degree of an unknown compound provided by the present disclosure is simple and easy to implement, and can provide a basis and convenience for the relevant development of preparations. DETAILED DESCRIPTION

[0040] In order to more clearly understand the above-mentioned objectives, features and advantages of the present disclosure, the scheme of the present disclosure will be further described below. It should be noted that the embodiments of the present disclosure and the features in the embodiments can be combined with each other without conflict.

[0041] In the following description, many specific details are set forth to facilitate a full understanding of the present disclosure, but the present disclosure may also be implemented in other ways different from those described herein; it is obvious that the embodiments in the specification are only part of the embodiments of the present disclosure, rather than all of the embodiments.

[0042] Example 1

[0043] This embodiment provides a model for determining the permeability of a drug to be tested, wherein the model includes a tool drug and a permeability parameter corresponding to the tool drug, as shown in Table 1:

[0044] Table 1

[0045]

[0046] Example 2

[0047] This example provides a method for determining the permeation degree of a drug to be tested using the model provided in Example 1.

[0048] (1) Determine the effective permeation rate of the drug to be tested using parallel artificial membrane permeation technology;

[0049] (2) comparing the effective permeation rate of the drug to be tested with the effective permeation rate of minoxidil to confirm whether the drug to be tested is a hyperosmotic drug;

[0050] (3) If the drug to be tested is a hyperosmotic drug, further compare it with the permeability parameters corresponding to the tool drug in the model to determine the permeability degree of the drug to be tested (high permeability high interval, high permeability middle interval and high permeability low interval):

[0051] The permeability parameter of the drug to be tested corresponds to pH=5.0, and the method comprises:

[0052] If the permeability of the drug to be tested is greater than the permeability of carbamazepine, the permeability of the drug to be tested is in the high permeability high interval;

[0053] If the permeability of the carbamazepine is greater than the permeability of the drug to be tested and greater than the permeability of the propranolol hydrochloride, the permeability of the drug to be tested is in the high permeability range;

[0054] If the permeability of the propranolol hydrochloride>the permeability of the drug to be tested>the permeability of the minoxidil, the permeability of the drug to be tested is in the high permeability low range.

[0055] The permeability parameter of the drug to be tested corresponds to pH=6.5, and the method comprises:

[0056] If the permeability of the drug to be tested is greater than the permeability of carbamazepine, the permeability of the drug to be tested is in the high permeability high interval;

[0057] If the permeability of the carbamazepine is greater than the permeability of the drug to be tested and greater than the permeability of the metoprolol tartrate, then the permeability of the drug to be tested is in the high permeability range;

[0058] If the permeability of the metoprolol tartrate>the permeability of the drug to be tested>the permeability of the minoxidil, the permeability of the drug to be tested is in the high permeability low range.

[0059] The permeability parameter of the drug to be tested corresponds to pH=7.4, and the method comprises:

[0060] If the permeability of the drug to be tested is greater than the permeability of propranolol hydrochloride, the permeability of the drug to be tested is in the high permeability high range;

[0061] If the permeability of the propranolol hydrochloride>the permeability of the drug to be tested>the permeability of phenytoin, then the permeability of the drug to be tested is in the high permeability middle range;

[0062] If the permeability of phenytoin>the permeability of the drug to be tested>the permeability of minoxidil, the permeability of the drug to be tested is in the high permeability low range.

[0063] Performance Test 1: Verifying the Accuracy of the Model Using the Known Hyperosmotic Drug Cilostazol

[0064] (1) The effective permeation rate of cilostazol was determined by parallel artificial membrane permeation technology:

[0065] A. Determination of standard curve:

[0066] The medium in the supply cell is pH=6.5, with a 10 mm optical path, a linear concentration range of 0.000-10 μg / mL, and a linear R 2 =0.9999; pH=7.4ABS solution in the receiving pool, 20mm optical path, linear concentration range 0-1.5μg / mL, linear R 2 =0.9999;

[0067] B. Drug permeability was measured using the μFlux device from pion.

[0068] Precisely measure 16 mL of dissolution medium (pH = 7.4) into the supply tank and 16 mL of ASB buffer (pH = 7.4) into the receiving tank. The speed is 200 rpm. The area between the two compartments is 1.65 cm. 2 The artificial membrane (infiltrated with 20 μL gastrointestinal simulated liposome solution) was used for isolation. The optical fibers were placed in each supply pool and receiving pool respectively, and the supply pool was water bathed at 37°C. The drug concentration in the supply pool was 50 μg / mL. The optical fiber specification was 10 mm, and the wavelength range was 314-324 nm. The effective permeability was calculated by the following formula;

[0069] Pe=[(dC / dt)×V] / (A×Ct×60)

[0070] Where Pe is the effective permeability, cm·s -1 ; dC / dt is the rate of change of the concentration of the drug to be tested in the receiving pool per unit time, μg·mL -1 ·min -1; V is the volume of buffer in the receiving pool corresponding to the change rate, mL; A is the membrane area, cm 2 ; Ct is the initial concentration of the drug to be tested in the supply pool, μg / mL;

[0071] Calculated from the above formula, Pe (effective permeability coefficient) = 274 × 10 -6 cm / s;

[0072] (2) Confirmation of the penetration of cilostazol

[0073] The effective permeation rate of cilostazol at pH = 6.5 was compared with the permeability parameters of the tool drug at pH = 6.5, and it was found that:

[0074] The effective permeation rate of cilostazol (274×10 -6 cm / s)>effective penetration rate of minoxidil (8.36×10 -6 cm / s);

[0075] Therefore, cilostazol was judged to be a hyperosmotic drug;

[0076] (3) Further confirmation of the penetration level of cilostazol in hyperosmotic drugs

[0077] Comparing the effective permeation rate of cilostazol with the permeability parameters of the tool drug at pH = 6.5, it was found that:

[0078] The effective permeation rate of cilostazol (274×10 -6 cm / s)>effective permeation rate of propranolol hydrochloride (245×10 -6 cm / s);

[0079] Therefore, the permeation degree of cilostazol was determined to be in the high permeability high range.

[0080] From the above verification, it can be seen that the model provided by the present disclosure can accurately attribute the penetration degree of unknown compounds and meet the application in the formulation research and development process.

[0081] Performance Test 2

[0082] The absorption of propranolol, milodinil, phenytoin, carbamazepine, and metoprolol in different intestinal segments of SD rats was investigated as follows:

[0083] (1) Animal experiments

[0084] Rats were fasted for 12 hours before the experiment, and they were allowed to drink water freely during the fasting period. Rats were anesthetized by ip administration of ulanose (20 mg / mL, 0.5 mL for every 100 g of body weight of rats), fixed, and a small incision was made along the midline of the abdomen to expose the rat intestine. The two ends of the small intestine were intubated and ligated; the intestinal contents were drained with physiological saline preheated to 37°C, replaced with blank KR solution for 15 minutes, and then replaced with drug-containing intestinal perfusion solution to saturate the pipeline for 15 minutes; the perfusion volume flow rate was reduced to 0.3 mL / min, and the perfusion solution and collection bottle were replaced once every 15 minutes. The experiment lasted for 120 minutes. After the experiment, the rats were killed, the perfused intestinal segments were cut, and the intestinal segments were cut open under 37°C physiological saline infiltration, and their length (L) and cross-sectional radius (r) were measured;

[0085] Each intestinal segment is divided into the duodenum segment: 10 cm from 1 cm below the pylorus; the jejunum segment: 10 cm from 15 cm below the pylorus; and the ileum segment: 10 cm from 20 cm above the cecum. Carbamazepine and metoprolol intestinal perfusion solution (mass concentration 40 μg / mL), propranolol, milodinil, and phenytoin perfusion solution concentration (mass concentration of about 20 μg / mL) were used to perform in vivo unidirectional intestinal perfusion experiments on each intestinal segment according to the above method to investigate the effects of different intestinal segments on drug absorption.

[0086] (2) Sample processing

[0087] Take 100 μL of the collected liquid sample, add 300 μL of internal standard (aspirin, 10 μg / mL) methanol, shake for 60 s, then centrifuge at 13000 rpm / min for 10 min, aspirate 100 μL of supernatant, and perform HPLC-MS / MS analysis.

[0088] (3) Results analysis

[0089] According to Papp < ​​1.8 × 10 -4 cm / min or Papp>1.2×10 -3 cm / min, respectively, indicating that the drug is poorly absorbed or completely absorbed (high permeability). In the test analysis, it was found that:

[0090] a. All five drugs are absorbed in the duodenum, jejunum, and ileum;

[0091] b. In the duodenum (pH = 5.0), the permeation rates of the five drugs were ranked as follows: minoxidil < metoprolol tartrate < propranolol hydrochloride < phenytoin < carbamazepine;

[0092] c. Minoxidil can be completely absorbed in the duodenum (pH = 5.0);

[0093] From the above, it can be seen that the tool drugs included in the model provided by the present disclosure are all hyperosmotic drugs. At the same time, the permeation rate in the duodenal (pH = 5) environment is ranked the same as the model provided by the present disclosure, which shows that the model provided by the present disclosure can accurately attribute the permeability of unknown compounds and meet the application in the formulation development process.

[0094] It should be noted that, in this article, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0095] The above description is only a specific embodiment of the present disclosure, so that those skilled in the art can understand or implement the present disclosure. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present disclosure. Therefore, the present disclosure will not be limited to the embodiments described herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A model for determining the penetration degree of a drug to be tested, characterized in that The model includes a tool drug and a permeability parameter corresponding to the tool drug; The tool drugs include carbamazepine, propranolol hydrochloride, metoprolol tartrate, phenytoin and minoxidil; The permeability parameters corresponding to the tool drug include effective permeation rate; The effective permeation rate includes the effective permeation rate corresponding to pH=5.0, the effective permeation rate corresponding to pH=6.5 and the effective permeation rate corresponding to pH=7.4; The effective permeation rate is determined by a parallel artificial membrane in vitro permeation model; The classification of the permeability degree includes high permeability, and the high permeability includes a high permeability high interval, a high permeability medium interval, and a high permeability low interval; The method for determining the penetration degree of the drug to be tested is as follows: If the permeability of the drug to be tested is greater than the permeability of minoxidil, the permeability of the drug to be tested is high permeability; The permeability of the drug to be tested is high permeability, and a method for further determining the permeability of the drug to be tested includes: The pH value corresponding to the permeability parameter corresponding to the drug to be tested is determined, and then compared with the permeability parameter corresponding to the same pH value of the tool drug to further determine whether the drug to be tested belongs to the high permeability high interval, the high permeability medium interval or the high permeability low interval.

2. Use of the model described in claim 1 in determining the penetration degree of a drug to be tested.

3. A method for determining the penetration degree of a drug to be tested using the model of claim 1, characterized in that: The method comprises: (1) Determine the permeability parameters corresponding to the drug to be tested; (2) comparing the permeability parameter corresponding to the drug to be tested with the permeability parameter corresponding to the tool drug included in the model of claim 1, thereby determining the degree of permeation of the drug to be tested; The method for determining the penetration degree of the drug to be tested is as follows: If the permeability of the drug to be tested is greater than the permeability of minoxidil, the permeability of the drug to be tested is high permeability; The permeability of the drug to be tested is high permeability, and a method for further determining the permeability of the drug to be tested includes: The pH value corresponding to the permeability parameter corresponding to the drug to be tested is determined, and then compared with the permeability parameter corresponding to the same pH value of the tool drug to further determine whether the drug to be tested belongs to the high permeability high interval, the high permeability medium interval or the high permeability low interval.

4. The method according to claim 3, characterized in that The permeability parameter of the drug to be tested corresponds to pH=5.0, and the method for further determining the permeability of the drug to be tested includes: If the permeability of the drug to be tested is greater than the permeability of carbamazepine, the permeability of the drug to be tested is in the high permeability high interval; If the permeability of the carbamazepine is greater than the permeability of the drug to be tested and greater than the permeability of the propranolol hydrochloride, the permeability of the drug to be tested is in the high permeability range; If the permeability of the propranolol hydrochloride>the permeability of the drug to be tested>the permeability of the minoxidil, the permeability of the drug to be tested is in the high permeability low range.

5. The method according to claim 3, characterized in that: The permeability parameter of the drug to be tested corresponds to pH=6.5, and the method for further determining the permeability of the drug to be tested includes: If the permeability of the drug to be tested is greater than the permeability of carbamazepine, the permeability of the drug to be tested is in the high permeability high interval; If the permeability of the carbamazepine is greater than the permeability of the drug to be tested and greater than the permeability of the metoprolol tartrate, then the permeability of the drug to be tested is in the high permeability range; If the permeability of the metoprolol tartrate>the permeability of the drug to be tested>the permeability of the minoxidil, the permeability of the drug to be tested is in the high permeability low range.

6. The method according to claim 3, characterized in that The permeability parameter of the drug to be tested corresponds to pH=7.4, and the method for further determining the permeability of the drug to be tested includes: If the permeability of the drug to be tested is greater than the permeability of propranolol hydrochloride, the permeability of the drug to be tested is in the high permeability high range; If the permeability of the propranolol hydrochloride>the permeability of the drug to be tested>the permeability of phenytoin, then the permeability of the drug to be tested is in the high permeability middle range; If the permeability of phenytoin>the permeability of the drug to be tested>the permeability of minoxidil, the permeability of the drug to be tested is in the high permeability low range.

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