Palmitoyl hydrochloride-indomethacin pharmaceutical co-crystal as well as preparation method and application thereof

Abstract

The application discloses a palmitoyl hydrochloride-indometacin pharmaceutical cocrystal as well as a preparation method and application thereof, and belongs to the technical field of pharmaceutical crystallization. 21 H 22 ClNO4]·[C 19 H 16 ClNO4]. The palmitoyl hydrochloride-indometacin pharmaceutical cocrystal is prepared by mixing palmitoyl hydrochloride hydrate and indometacin in a solvent system according to a molar ratio of 1:1 and through stirring treatment in a suspended state. The process flow is simple, the operation is convenient, and the cocrystal has a high yield. In a pH 6.8 simulated intestinal fluid medium, the dissolution rate of palmitoyl hydrochloride in the pharmaceutical cocrystal is obviously lower than that of pure palmitoyl hydrochloride, and the pharmaceutical cocrystal presents a sustained-release behavior. On the contrary, the dissolution rate of indometacin in the pharmaceutical cocrystal is obviously improved, which is beneficial to the improvement of the bioavailability of indometacin. The palmitoyl hydrochloride-indometacin pharmaceutical cocrystal also has good human colon cancer cell inhibition activity.

Description

Technical Field

[0001] This invention belongs to the field of drug crystallization technology, and specifically relates to a method for preparing drug-drug cocrystals and their applications. Background Technology

[0002] Palmitine chloride, also known as berberine, possesses well-defined antibacterial and anti-inflammatory properties. Clinically, it is commonly used to treat bacterial dysentery, enteritis, and respiratory and urinary tract infections. It can also be used to treat gynecological inflammation and conjunctivitis. In recent years, research has delved deeper into its anti-inflammatory and antioxidant properties and systematically elucidated its mechanism of action. Furthermore, this compound exhibits various pharmacological activities, including antitumor, hypoglycemic, and anti-Alzheimer's disease effects.

[0003] Indomethacin, also known as anti-inflammatory drug, is a classic nonsteroidal anti-inflammatory drug (NSAID). It primarily works by inhibiting the activity of cyclooxygenase (COX) in the body, thereby reducing prostaglandin production and exerting antipyretic, analgesic, and anti-inflammatory effects. Clinically, it is widely used to treat arthritis, acute gout attacks, and soft tissue inflammation such as bursitis and tenosynovitis. Studies have also shown that it has some adjuvant effects in the field of anti-tumor therapy and a certain preventive effect against colorectal cancer. However, long-term use of indomethacin can easily cause gastrointestinal adverse reactions, such as gastric perforation and gastrointestinal bleeding. Current technologies mainly employ time-controlled drug release or enzyme-degraded microparticle strategies.

[0004] For example, the invention patent with publication number CN108743561A discloses an indomethacin colon-targeted capsule, which utilizes the specific degradation of guar gum in the colon to achieve targeted drug release. However, its preparation process is complex, and it is only a single drug formulation, which cannot achieve synergistic treatment. Another example is the invention patent with publication number CN109806238A, which discloses an indomethacin colon-release microsphere. It uses enzyme degrading agents such as pectin combined with hydrophobic blocking agents to prepare gel microspheres by a drop method. It also has the defects of complicated preparation process, limited drug loading capacity, and only containing a single active ingredient.

[0005] Drug-drug cocrystals, as a novel form of solid-state drug, assemble two or more active drug molecules into a single crystal structure through non-covalent interactions, exhibiting characteristics such as well-defined crystal structure and stable physicochemical properties. Cocrystallization strategies can not only optimize drug solubility, dissolution behavior, and stability but also achieve synergistic release and enhanced efficacy of multiple drugs. For example, invention patent CN118239854A discloses a mesalazine-tetracycline hydrochloride drug cocrystal and its preparation method. This cocrystal shows significantly reduced solubility in simulated gastric juice and exhibits sustained-release behavior in simulated intestinal juice, increasing the contact time between the drug and the inflamed intestinal mucosa and demonstrating certain synergistic anticancer activity. However, its half-maximal inhibitory concentration (IC50) against HT29 colorectal cancer cells is limited. 50 The concentration is approximately 60 μg / mL, which is still far from clinical application.

[0006] Therefore, developing a novel drug combination that can achieve synergistic effects between palmitate hydrochloride and indomethacin while reducing adverse reactions is of great significance for improving the treatment efficacy of inflammatory diseases and expanding their clinical application value. Summary of the Invention

[0007] This invention addresses the technical problems of limited efficacy of existing single-drug therapy and insufficient therapeutic activity of existing cocrystal drugs by proposing a palmitate-indomethacin cocrystal, its preparation method, and its application.

[0008] To achieve the above objectives, the technical solution of the present invention is implemented as follows:

[0009] In a first aspect, the present invention provides a palmitate hydrochloride-indomethacin drug cocrystal, formed by combining palmitate hydrochloride and indomethacin in a molar ratio of 1:1, with the molecular formula [C 21 H 22 ClNO4]·[C 19 H 16 ClNO4].

[0010] The X-ray diffraction pattern of the palmitate-indomethacin eutectic powder has characteristic peaks at the following positions within a diffraction angle of 2θ±0.3°: 9.1°, 10.6°, 16.8°, 17.0°, 18.0°, 18.6°, 20.1°, 20.8°, 22.3°, 23.3°, 24.8°, 25.5°, 26.6°, 27.1°, 29.8°, 34.2°, and 44.5°.

[0011] This invention provides a method for preparing the aforementioned palmitate-indomethacin drug cocrystal, comprising the following steps:

[0012] (1) The palmitate hydrochloride hydrate and indomethacin were added to a solvent for dispersion to obtain a uniform suspension system;

[0013] (2) After sealing the system, stir to carry out the reaction. After the reaction is completed, separate the obtained solid and dry it to obtain a palmitate-indomethacin drug cocrystal.

[0014] In step (1), the molar ratio of palmitate hydrochloride hydrate to indomethacin is 1:1, the initial concentration of indomethacin is 0.025-0.05 mol / L, and the solvent is ethanol.

[0015] In step (2), the stirring speed is 100-500 rpm, the reaction temperature is 10-30 ℃, and the time is 4-48 h; the drying temperature is 40-60 ℃, and the time is 2-12 h.

[0016] This invention provides the application of the aforementioned palmitate hydrochloride-indomethacin drug cocrystal in the preparation of antipyretic, analgesic, antibacterial, anti-inflammatory, and anticancer drugs.

[0017] This invention provides the use of the aforementioned palmitate hydrochloride-indomethacin cocrystal in the preparation of pharmaceutical compositions for treating urinary tract infections, gynecological inflammations, conjunctivitis, rheumatoid arthritis, osteoarthritis, and colon cancer.

[0018] The present invention also provides the application of the aforementioned palmitate-indomethacin drug cocrystal in the preparation of anti-colorectal cancer drugs.

[0019] In a second aspect, the present invention provides a pharmaceutical composition comprising an effective amount of palmitate hydrochloride-indomethacin pharmaceutical cocrystal, and a pharmaceutically acceptable carrier.

[0020] The content of the palmitate hydrochloride-indomethacin drug eutectic in the drug composition is 20-100% of the total mass of the drug composition.

[0021] The dosage form of the pharmaceutical composition is selected from tablets, capsules, granules, injections, suppositories, or sprays.

[0022] The beneficial effects of this invention are:

[0023] 1. This invention successfully constructed a palmitate hydrochloride-indomethacin drug cocrystal system for the first time. The preparation process is simple and mild, yielding a high cocrystal yield, and simultaneously introducing two active ingredients, palmitate hydrochloride and indomethacin, into the same crystal. Dynamic vapor adsorption (DVS) tests showed that the hygroscopic tendency of this drug cocrystal was significantly reduced compared to palmitate hydrochloride hydrate. At adsorption equilibrium at 25°C and 95% relative humidity, the water absorption mass fraction of the cocrystal was only 0.51%, indicating low hygroscopicity. Stability tests showed that it possesses good thermal stability, humidity stability, and light stability.

[0024] 2. In vitro dissolution behavior studies showed that in a simulated intestinal fluid environment at pH 6.8, palmitate hydrochloride in the cocrystal exhibited a sustained-release characteristic compared to the active pharmaceutical ingredient (API). Furthermore, the release rates of the two active components were relatively similar, contributing to a synergistic therapeutic effect. In vitro cell experiments indicated that in the HT-29 human colorectal cancer cell model, the IC50 values ​​of palmitate hydrochloride-indomethacin cocrystal, palmitate hydrochloride, and indomethacin were relatively high. 50 The values ​​were 12.2 μg / mL, 30.7 μg / mL, and 125.0 μg / mL, respectively, indicating that the two drugs produced a significant synergistic effect in the co-crystal state. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0026] Figure 1 The powder X-ray diffraction pattern of the palmitate-indomethacin drug eutectic prepared in Example 1 is shown.

[0027] Figure 2 The image shows the dynamic water vapor adsorption of the palmitate-indomethacin drug eutectic prepared in Example 1 at 25°C.

[0028] Figure 3 The results of the crystal stability test of the palmitate-indomethacin drug cocrystal prepared in Example 1 are shown.

[0029] Figure 4 The results are from an in vitro dissolution experiment of palmitate hydrochloride-indomethacin drug cocrystal. Detailed Implementation

[0030] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] Example 1

[0032] A method for preparing a palmitate hydrochloride-indomethacin drug cocrystal, the specific steps of which are as follows:

[0033] (1) 0.5 mmol palmitate hydrochloride hydrate and 0.5 mmol indomethacin powder were added to 10 mL of ethanol for dispersion to obtain a uniform suspension system;

[0034] (2) After sealing the above suspension system, stir at 300 rpm for 24 h at room temperature. After the reaction is completed, separate the precipitate and dry it at 50°C for 8 h to obtain palmitate hydrochloride-indomethacin drug cocrystal.

[0035] The palmitate hydrochloride-indomethacin drug cocrystal prepared in this embodiment was subjected to powder X-ray diffraction analysis, and the results are as follows: Figure 1 As shown, the prepared palmitate hydrochloride-indomethacin drug cocrystal exhibits characteristic diffraction peaks at diffraction angles of 2θ ± 0.3° at the following locations: 9.1, 10.6, 16.8, 17.0, 18.0, 18.6, 20.1, 20.8, 22.3, 23.3, 24.8, 25.5, 26.6, 27.1, 29.8, 34.2, and 44.5°.

[0036] Dynamic water vapor adsorption tests were performed on the palmitate-indomethacin cocrystal prepared in this embodiment. Figure 2 This is a dynamic water vapor adsorption diagram (SMS DVS Intrinsic) of palmitate hydrochloride hydrate and the palmitate hydrochloride-indomethacin cocrystal prepared in this example at 25°C. As shown in the diagram, during the process of increasing relative humidity from 0% RH to 95% RH, the mass change of palmitate hydrochloride hydrate is approximately 18%, while the mass change of the palmitate hydrochloride-indomethacin cocrystal is only 0.51%. This indicates that the hygroscopicity of the palmitate hydrochloride-indomethacin cocrystal is significantly reduced compared to that of palmitate hydrochloride hydrate.

[0037] The crystal stability of the palmitate-indomethacin cocrystal prepared in this embodiment was tested, and the results are as follows: Figure 3As shown, the palmitate hydrochloride-indomethacin cocrystal did not undergo any crystal form change when placed at 60℃ (protected from light) for 30 days, or at 95% RH (temperature 25℃, protected from light) for 30 days, or at a light intensity of 4500 Lx (temperature 25℃) for 30 days. This indicates that the cocrystal has good thermal stability, light stability, and humidity stability.

[0038] Example 2

[0039] A method for preparing a palmitate hydrochloride-indomethacin drug cocrystal, the specific steps of which are as follows:

[0040] (1) 0.5 mmol palmitate hydrochloride hydrate and 0.5 mmol indomethacin powder were added to 15 mL of ethanol for dispersion to obtain a uniform suspension system;

[0041] (2) After sealing the above suspension system, stir at 100 rpm for 48 h at 10 °C. After the reaction is completed, separate the precipitate and then dry it at 60 °C for 2 h to obtain palmitate hydrochloride-indomethacin drug cocrystal.

[0042] Example 3

[0043] A method for preparing a palmitate hydrochloride-indomethacin drug cocrystal, the specific steps of which are as follows:

[0044] (1) 0.5 mmol palmitate hydrochloride hydrate and 0.5 mmol indomethacin powder were added to 20 mL of ethanol for dispersion to obtain a uniform suspension system;

[0045] (2) After sealing the above suspension system, stir at 500 rpm at 30°C for 4 h. After the reaction is completed, separate the precipitate and dry it at 40°C for 12 h to obtain palmitate hydrochloride-indomethacin drug cocrystal.

[0046] Example of implementation effect 1

[0047] An in vitro dissolution experiment was conducted on the palmitate-indomethacin cocrystal prepared in Example 1, as follows:

[0048] Palmitin hydrochloride-indomethacin cocrystal and indomethacin powder were sieved through a 100-mesh sieve to ensure uniform particle size and reduce the influence of particle size on dissolution. The experiment employed a small-cup paddle method with a paddle speed of 75 rpm / min, using 250 mL of pH 6.8 simulated intestinal fluid as the dissolution medium. Samples of 96.1 mg of palmitin hydrochloride-indomethacin cocrystal, 46.1 mg of indomethacin, and 57.0 mg of palmitin hydrochloride hydrate were weighed separately. After the water temperature stabilized at 37℃, the samples were added to the pH 6.8 simulated intestinal fluid dissolution medium. At specific time points, 1 mL of solution was taken, and 1 mL of medium solution was immediately added. All extracted solutions were filtered through a 0.45 μm membrane, and their concentrations were measured by HPLC. Three parallel samples were used (n = 3). The results are as follows: Figure 4 As shown.

[0049] Depend on Figure 4 It can be seen that in the pH 6.8 simulated intestinal fluid dissolution medium, compared with palmitate hydrochloride hydrate and indomethacin, the dissolution rate of palmitate hydrochloride in the palmitate hydrochloride-indomethacin cocrystal is significantly reduced, while the dissolution rate of indomethacin is significantly increased. The difference in dissolution rate between the two active ingredients is significantly reduced, and they are closer to synchronous release, which is conducive to achieving better therapeutic effects.

[0050] Example of implementation effect 2

[0051] The palmitate-indomethacin cocrystal prepared in Example 1 was subjected to an inhibitory effect on the proliferation of human colorectal cancer cells, as detailed below:

[0052] Tumor cell inhibition was determined by the MTT assay. HT-29 human colorectal cancer cells in logarithmic growth phase were seeded into 96-well culture plates at a density of 3000 cells / 100 μL per well. After 24 h of culture, 100 μL of different concentrations of sample solution were added to the experimental groups, with each concentration used in triplicate; 100 μL of nutrient solution was added to the negative control group and 100 μL of nutrient solution was added to the blank control wells for instrument zeroing. After 72 h of treatment, the culture medium was removed, and 100 µL of 10% trichloroacetic acid (TCA) was added to each well. The cells were fixed overnight at 4°C. After removing the TCA, the cells were washed 5 times with double-distilled water. 100 μL of sulfonylrhodamine B (SRB) staining solution was added per well. After 0.5 h, the cells were washed 3-4 times with 0.01% acetic acid solution. After air drying, 100 μL of 0.1M Tris solution was added to each well. The cells were shaken for approximately 5 min until the crystals in the cells were fully dissolved. The absorbance of each well at 550 nm was measured using a microplate reader. The average values ​​of each group were taken, and the cell viability of the negative control group was set at 100%. The cell proliferation inhibition rate (IR) of each group was calculated using the formula: Inhibition rate = (1 - A550 value of experimental group / A550 value of control group) × 100%. A dose-response curve was plotted with logarithmic concentration on the x-axis and inhibition rate on the y-axis. Linear regression was performed on the inhibition rates between 20% and 80% to calculate the IC50. 50 The results are shown in Table 1:

[0053] Table 1. IC50 of palmitate hydrochloride hydrate, indomethacin, and the palmitate hydrochloride-indomethacin cocrystal prepared in Example 3 against HT29 human colorectal cancer cells. 50

[0054]

[0055] Table 1 shows that in the HT-29 human colorectal cancer cell model, the IC50 values ​​of palmitate-indomethacin cocrystal, palmitate hydrochloride, and indomethacin were... 50 The values ​​were 12.2 μg / mL, 30.7 μg / mL, and 125.0 μg / mL, respectively, indicating that the two drugs produced a significant synergistic effect in the co-crystal state.

[0056] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A palmitate hydrochloride-indomethacin drug cocrystal, characterized in that: It is formed by the covalent bonding of palmitate hydrochloride and indomethacin, with the molecular formula [C]. 21 H 22 ClNO4]·[C 19 H 16 [ClNO4], wherein the molar ratio of palmitate hydrochloride to indomethacin is 1:

1.

2. The palmitate-indomethacin drug eutectic according to claim 1, characterized in that: The X-ray diffraction pattern of the palmitate-indomethacin eutectic powder has characteristic peaks at the following positions within a diffraction angle of 2θ±0.3°: 9.1°, 10.6°, 16.8°, 17.0°, 18.0°, 18.6°, 20.1°, 20.8°, 22.3°, 23.3°, 24.8°, 25.5°, 26.6°, 27.1°, 29.8°, 34.2°, and 44.5°.

3. The method for preparing the palmitate-indomethacin drug cocrystal according to claim 1 or 2, characterized in that, Includes the following steps: (1) The palmitate hydrochloride hydrate and indomethacin were added to a solvent for dispersion to obtain a uniform suspension system; (2) After sealing the system, stir to carry out the reaction. After the reaction is completed, separate the obtained solid and dry it to obtain a palmitate-indomethacin drug cocrystal.

4. The preparation method according to claim 3, characterized in that: In step (1), the molar ratio of palmitate hydrochloride hydrate to indomethacin is 1:1, and the initial concentration of indomethacin is 0.025-0.05 mol / L; the solvent is ethanol.

5. The preparation method according to claim 4, characterized in that: In step (2), the stirring speed is 100-500 rpm, the reaction temperature is 10-30 ℃, and the time is 4-48 h; the drying temperature is 40-60 ℃, and the time is 2-12 h.

6. The use of the palmitate hydrochloride-indomethacin drug eutectic according to claim 1 or 2 in the preparation of antipyretic, analgesic, antibacterial, anti-inflammatory and anticancer drugs.

7. The use of the palmitate-indomethacin drug eutectic according to claim 1 or 2 in the preparation of anti-colorectal cancer drugs.

8. A pharmaceutical composition, characterized in that: The product comprises an effective amount of the palmitate-indomethacin pharmaceutical cocrystal of claim 1 or 2, and a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to claim 8, characterized in that: The content of the palmitate hydrochloride-indomethacin drug eutectic is 20-100% of the total mass of the drug composition.

10. The pharmaceutical composition according to claim 9, characterized in that: The dosage form of the pharmaceutical composition is selected from tablets, capsules, granules, injections, suppositories, or sprays.

Images