Pharmaceutical composition containing imatinib as well as preparation method and application thereof

Abstract

The invention provides new application of piperazine compounds or compositions thereof, namely preparation of ophthalmic drugs for preventing and / or treating alkali burns, as well as pharmaceutical compositions and preparation methods of the piperazine compounds or the compositions thereof. The piperazine compound (imatinib or pharmaceutically acceptable salt thereof) and the composition thereof have a good curative effect on prevention and / or treatment of eye chemical injury (especially alkali burn), are non-irritant, and can accelerate healing of corneal injury, improve corneal sensitivity, inhibit neovascularization caused by the eye chemical injury and the like; test results further show that the piperazine compound and the glycyrrhizinate are combined, so that the piperazine compound and the glycyrrhizinate are high in safety, good in stability and easy to dissolve, release and / or absorb, the bioavailability of active drugs is improved, the piperazine compound and the glycyrrhizinate have a synergistic effect, a treatment effect better than that of any single component is achieved, and the piperazine compound and the glycyrrhizinate have a wide application prospect.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to a pharmaceutical composition containing imatinib, a preparation method thereof, and uses thereof. Background Art

[0002] The cornea is an important barrier for the eye to resist external microorganisms and bacteria. Since the cornea is directly exposed to the outside world, it is easily damaged by various factors such as mechanical, infectious, chemical, and burn. The health of the cornea affects the health status of the eyeball. Timely and effective repair of the damaged cornea can prevent the development and deterioration of the condition.

[0003] Ocular chemical burns, as a common ophthalmic emergency of non-mechanical injury, mostly occur in chemical plants, construction sites, laboratories, etc. According to different chemical substances, they can be divided into alkali burns, acid burns, etc. Among them, alkali burns are often more serious than acid burns.

[0004] After the eye suffers chemical injury, eye tissues such as the cornea are damaged and inflammation is triggered. If the inflammatory reaction cannot be inhibited and treated in time, it may further lead to necrosis of eye tissues, thus resulting in a vicious cycle. At present, the treatment means for such ocular diseases are very limited. In addition to drug treatment, severe patients often need corneal transplantation and even have the risk of blindness.

[0005] Imatinib belongs to piperazine compounds (compounds containing a piperazine structure in the compound structure), and is a small molecule protein kinase inhibitor. It has the effect of blocking one or more protein kinases. Clinically, it is mainly used in the form of its salts to treat chronic myeloid leukemia, malignant gastrointestinal stromal tumors, etc. For example, imatinib mesylate capsules, etc.

[0006] Dipotassium glycyrrhizinate (DG) is a white or off-white powder, with anti-inflammatory, anti-allergic, moisturizing and other effects. In the pharmaceutical industry, it is mainly used for relieving cough and reducing phlegm, gastric ulcer, acute and chronic gastritis, eczema, skin itching, and for treating cancer and preventing and treating AIDS, etc.

[0007] So far, no relevant literature reports have been retrieved on using imatinib or its combination with dipotassium glycyrrhizinate for preventing, treating and / or relieving ocular chemical burns (such as alkali burns).

[0008] In view of this, the present invention is specifically proposed. Summary of the Invention

[0009] In view of the problems and / or deficiencies existing in the prior art, one of the objectives of the present invention is to provide a new use of piperazine compounds (imatinib or its pharmaceutically acceptable salts) or their compositions: preparing ophthalmic drugs for preventing and / or treating alkali burns. The piperazine compounds (imatinib or its pharmaceutically acceptable salts) and their compositions of the present invention, as drugs for preventing and / or treating ocular chemical injuries, can accelerate the healing of corneal injuries, improve corneal sensitivity, inhibit neovascularization caused by ocular chemical injuries, etc., have good curative effects, and are safe and non-irritating.

[0010] The present invention provides a new use:

[0011] Use of a piperazine compound or its composition in the preparation of an ophthalmic drug for preventing and / or treating alkali burns, wherein the composition comprises a piperazine compound and a glycyrrhizinate; the piperazine compound is imatinib or its pharmaceutically acceptable salt; the glycyrrhizinate is selected from one or more of sodium glycyrrhizinate, disodium glycyrrhizinate, potassium glycyrrhizinate, dipotassium glycyrrhizinate, ammonium glycyrrhizinate, and diammonium glycyrrhizinate;

[0012] Preferably, the piperazine compound is imatinib; and / or the glycyrrhizinate is dipotassium glycyrrhizinate or disodium glycyrrhizinate.

[0013] Furthermore,

[0014] In any of the above technical solutions (use in the preparation of an ophthalmic drug for preventing and / or treating alkali burns), the solvent of the piperazine compound or its composition is water or phosphate buffer solution, and the ophthalmic drug is eye drops.

[0015] Furthermore,

[0016] In any of the above technical solutions (use in the preparation of an ophthalmic drug for preventing and / or treating alkali burns), the solvent of the piperazine compound or its composition is water or phosphate buffer solution, the composition contains a cellulose derivative and a hyaluronic acid compound, and the ophthalmic drug is an ophthalmic gel;

[0017] wherein the cellulose derivative is selected from one or more of methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, calcium carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylcellulose, methyl ethylcellulose, hydroxyethylcellulose, and hydroxyethylmethylcellulose; the hyaluronic acid compound is hyaluronic acid or its salt;

[0018] Preferably, the cellulose derivative is hydroxypropylmethylcellulose; the hyaluronic acid compound is an alkali metal salt of hyaluronic acid (for example, sodium hyaluronate, potassium hyaluronate), and more preferably sodium hyaluronate.

[0019] The present invention also provides a pharmaceutical composition, comprising a piperazine compound and a pharmaceutically acceptable excipient; the piperazine compound is imatinib or a pharmaceutically acceptable salt thereof, and the pharmaceutically acceptable excipient comprises glycyrrhizinate;

[0020] Preferably, the mass ratio of the piperazine compound to the glycyrrhizinate is 1:1 to 20 (for example, the mass ratio is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, etc.);

[0021] and / or, the glycyrrhizinate is selected from one or more of sodium glycyrrhizinate, disodium glycyrrhizinate, potassium glycyrrhizinate, dipotassium glycyrrhizinate, ammonium glycyrrhizinate, and diammonium glycyrrhizinate;

[0022] and / or, the piperazine compound is imatinib;

[0023] More preferably,

[0024] the mass ratio of the piperazine compound to the glycyrrhizinate is 1:4 to 10, and further preferably the mass ratio of the piperazine compound to the glycyrrhizinate is 1:5.5 to 6.5;

[0025] and / or, the glycyrrhizinate is dipotassium glycyrrhizinate or disodium glycyrrhizinate.

[0026] Furthermore,

[0027] in any of the above technical solutions (pharmaceutical composition), the piperazine compound exists in the pharmaceutical composition in an amorphous form;

[0028] and / or, the encapsulation efficiency of the piperazine compound is at least 80% (for example, the encapsulation efficiency is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, etc.); preferably, the encapsulation efficiency of the piperazine compound is ≥90% or ≥95%.

[0029] Furthermore,

[0030] in any of the above technical solutions (pharmaceutical composition), the pharmaceutical composition is a liquid preparation, semi-solid preparation or solid preparation; and / or, the piperazine compound in the pharmaceutical composition is in a therapeutically effective amount.

[0031] Furthermore,

[0032] In any of the above technical solutions (pharmaceutical composition), the pharmaceutical composition is a liquid preparation, the solvent of the pharmaceutical composition is water or phosphate buffer solution, and the liquid preparation is an eye drop;

[0033] Preferably, when the concentration of the piperazine compound in the liquid preparation is 0.5 mg / mL, the liquid preparation satisfies one or more of the following conditions ① to ③:

[0034] ①. The average particle size of the liquid preparation is 1 to 100 nm (for example, the average micelle particle size is 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 100 nm, etc.), preferably 5 to 20 nm;

[0035] ②. The polydispersity coefficient of the liquid preparation is ≤0.6, preferably 0.1 to 0.5;

[0036] ③. The Zeta potential of the liquid preparation is -30 to -1 mV, preferably -25 to -15 mV;

[0037] More preferably, the liquid preparation simultaneously satisfies the above conditions ① to ③.

[0038] Furthermore,

[0039] In any of the above technical solutions (pharmaceutical composition), the pharmaceutical composition is a semi-solid preparation, the pharmaceutical composition contains a cellulose derivative and a hyaluronic acid compound, the solvent of the pharmaceutical composition is water or phosphate buffer solution, and the semi-solid preparation is an ophthalmic gel;

[0040] Preferably,

[0041] The mass ratio of the piperazine compound to the cellulose derivative is 1:1 to 20 (for example, the mass ratio is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, etc.);

[0042] And / or, the mass ratio of the piperazine compound and the hyaluronic acid compound is 1:0.5 to 20 (for example, the mass ratio is 1:0.5, 1:0.8, 1:1, 1:1.2, 1:1.5, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, etc.);

[0043] And / or, the cellulose derivative is selected from one or more of methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, methyl ethyl cellulose, hydroxyethyl cellulose, and hydroxyethyl methyl cellulose;

[0044] And / or, the hyaluronic acid compound is hyaluronic acid or its salt;

[0045] More preferably,

[0046] The mass ratio of the piperazine compound and the cellulose derivative is 1:2 to 5, and further preferably 1:2.5;

[0047] And / or, the mass ratio of the piperazine compound and the hyaluronic acid compound is 1:0.9 to 1.5, and further preferably 1:1;

[0048] And / or, the cellulose derivative is hydroxypropyl methyl cellulose;

[0049] And / or, the hyaluronic acid compound is an alkali metal salt of hyaluronic acid (for example, sodium hyaluronate, potassium hyaluronate), and further preferably sodium hyaluronate.

[0050] Furthermore,

[0051] In any of the above technical solutions (pharmaceutical composition), the pharmaceutical composition is a pharmaceutical composition for preventing and / or treating ocular chemical burns;

[0052] Preferably, the ocular chemical burn is an alkali burn.

[0053] Furthermore,

[0054] In any of the above technical solutions (drug composition), the drug composition is prepared by a method comprising the following steps: dissolving or dispersing the piperazine compound and the glycyrrhizinate in an organic solvent, and then removing the organic solvent to obtain a product; optionally including a formulation step: dissolving or dispersing the aforementioned product in the solvent of the drug composition, or dissolving or dispersing the aforementioned product together with the cellulose derivative and the hyaluronic acid compound in the solvent of the drug composition, thus obtaining the product;

[0055] Preferably, the organic solvent is an alcohol solvent; and / or, the amount of the organic solvent used per milligram of the piperazine compound is 0.1 - 20 mL (for example, the amount of the organic solvent used is 0.1 mL, 0.5 mL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL, 17 mL, 18 mL, 19 mL, 20 mL, etc.);

[0056] More preferably, the organic solvent is ethanol; and / or, the amount of the organic solvent used per milligram of the piperazine compound is 1 - 10 mL.

[0057] The present invention also provides a method for preparing the drug composition according to any one of the above, comprising the following steps: dissolving or dispersing the piperazine compound and the glycyrrhizinate in an organic solvent, and then removing the organic solvent to obtain a product; optionally including a formulation step: dissolving or dispersing the aforementioned product in the solvent of the drug composition, or dissolving or dispersing the aforementioned product together with the cellulose derivative and the hyaluronic acid compound in the solvent of the drug composition, thus obtaining the product;

[0058] Preferably, the organic solvent is an alcohol solvent; and / or, the amount of the organic solvent used per milligram of the piperazine compound is 0.1 - 20 mL (for example, the amount of the organic solvent used is 0.1 mL, 0.5 mL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL, 17 mL, 18 mL, 19 mL, 20 mL, etc.);

[0059] More preferably, the organic solvent is ethanol; and / or, the amount of the organic solvent used per milligram of the piperazine compound is 1 - 10 mL.

[0060] Regarding the definitions of the terms used, unless otherwise specified, the initial definitions provided for the terms in this article apply to the term throughout the article; for terms not specifically defined in this article, their meanings should be given according to the disclosed content and / or context and the common knowledge in the art, which can be understood by those skilled in the art.

[0061] The term "pharmaceutically acceptable" means generally chemically or physically compatible with other components constituting a pharmaceutical dosage form and physiologically compatible with the receptor.

[0062] The term "excipient" refers to a substance contained in a dosage form other than the active ingredient.

[0063] The term "therapeutically effective amount" refers to the amount of a pharmaceutical compound administered to a patient that is sufficient to effectively treat a disease. The therapeutically effective amount will vary depending on the pharmaceutical compound, the type of disease, the severity of the disease, the age of the patient, etc., and can be routinely adjusted by those skilled in the art according to the circumstances.

[0064] The term "ophthalmic drug" refers to a drug used to treat and / or diagnose eye diseases in the form of eye washing, eye drops, etc. Ophthalmic drugs can be divided into ophthalmic liquid preparations (including but not limited to eye drops, eye washes, intraocular injection solutions, etc.), ophthalmic semi-solid preparations (including but not limited to eye ointments, ophthalmic creams, ophthalmic gels, etc.), ophthalmic solid preparations (including but not limited to eye films, eye pills, intraocular inserts, etc.), etc. According to different needs and / or administration forms, ophthalmic drugs may contain pH regulators (such as buffers), osmotic pressure regulators, preservatives, antibacterial agents, viscosity regulators, solubilizers, dispersants, cosolvents, antioxidants, etc. commonly used in the art.

[0065] The beneficial effects of the present invention mainly include the following aspects:

[0066] (1) The present invention provides a new use of piperazine compounds (imatinib or its pharmaceutically acceptable salts) or their compositions: the use in the preparation of ophthalmic drugs for preventing and / or treating alkali burns;

[0067] (2) The in vitro and / or in vivo test results show that the piperazine compounds (imatinib or its pharmaceutically acceptable salts) and their compositions of the present invention have excellent efficacy for preventing and / or treating ocular chemical burns (especially alkali burns), are non-irritating, can accelerate the healing of corneal injuries, improve corneal sensitivity, and inhibit neovascularization caused by ocular chemical burns;

[0068] (3) In vitro and / or in vivo test results further show that the combination of piperazine compounds (imatinib or its pharmaceutically acceptable salts) and glycyrrhizinate has high safety, good stability, is easy to dissolve, release, and / or absorb, improves the bioavailability of the active drug, and the two have a synergistic effect, achieving a more superior therapeutic effect than any single component. Description of the Drawings

[0069] Figure 1 It is a graph of the encapsulation efficiency of IMB-DG obtained with different ratios in Example 1.

[0070] Figure 2 It is a comparative FTIR spectrum of IMB-DG in Example 4.

[0071] Figure 3 It is a comparative XRD spectrum of IMB-DG in Example 4.

[0072] Figure 4 It is a comparative graph of the in vitro release curve of IMB in IMB-DG in Example 5.

[0073] Figure 5 It is an observation diagram of the irritation of the cornea and conjunctiva in Example 6.

[0074] Figure 6 It is a comparative graph of corneal fluorescein sodium staining of different administration groups in Example 7.

[0075] Figure 7 It is a comparative graph of the corneal epithelial area defect rate of different administration groups in Example 7.

[0076] Figure 8 It is a comparative graph of the corneal opacity score of different administration groups in Example 7.

[0077] Figure 9 It is a comparative graph of the corneal sensitivity of different administration groups in Example 7.

[0078] Figure 10 It is a comparative observation diagram of ocular neovascularization of different administration groups in Example 7.

[0079] Figure 11 It is a comparative graph of the corneal neovascularization score of different administration groups in Example 7. Detailed Description of the Invention

[0080] The present invention will be clearly and completely described below in conjunction with specific embodiments. Those skilled in the art will understand that the following embodiments are some embodiments of the present invention, rather than all embodiments, and are only used to illustrate the present invention and should not be regarded as limiting the protection scope of the present invention.

[0081] In the embodiments of the present invention, for the tests without specific conditions noted, they are carried out according to the conventional conditions in the art or the conditions recommended by the manufacturer. For the reagents and instruments without the manufacturer noted, they are all conventional products that can be obtained by purchasing in the market.

[0082] Imatinib (abbreviated as "IMB"): Purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0083] Dipotassium Glycyrrhizinate (abbreviated as "DG"): Purity ≥ 98%, purchased from Shaanxi Fujie Pharmaceutical Co., Ltd.

[0084] Phosphate Buffered Saline (PBS): Purchased from Wuhan Sevier Biotechnology Co., Ltd.

[0085] Male New Zealand white rabbits: Purchased from Qingdao Kangda Biotechnology Co., Ltd.

[0086] C57BL / 6J male mice: 8 weeks old, purchased from Jinan Pengyue Laboratory Animal Breeding Co., Ltd.

[0087] SPSS Statistics 26 software is used for data analysis. P < 0.05 indicates a significant difference in statistical significance.

[0088] Example 1

[0089] Dissolve or disperse 5 mg of imatinib (IMB) with different masses of dipotassium glycyrrhizinate (DG) (the mass ratios of IMB to DG are 1:1, 1:2, 1:3, 1:4, 1:5, 1:6 respectively) in ethanol (for example, 20 mL). After mixing evenly, use a rotary evaporator to evaporate ethanol at 40 °C to obtain the solid product IMB-DG.

[0090] 1.1 Encapsulation efficiency test

[0091] Dissolve the obtained IMB-DG with water (the concentration of imatinib is about 1 mg / mL), filter through a 0.22 μm filter membrane to separate the unencapsulated IMB. Dilute the solutions before and after filtering of IMB-DG with methanol to disrupt the micelles. Use a UV spectrophotometer to measure the absorbance OD value at the maximum absorption wavelength and calculate the encapsulation efficiency. The encapsulation efficiency is the ratio (%) of the OD value detected after filtering to the OD value detected before filtering.

[0092] The encapsulation efficiencies of IMB-DG obtained from the aforementioned different ratios (the mass ratios of IMB to DG are 1:1, 1:2, 1:3, 1:4, 1:5, 1:6 respectively) are as Figure 1As shown in the figure. The results showed that the encapsulation efficiencies measured when the mass ratio of IMB to DG was 1:4, 1:5, and 1:6 were 85.23±2.56%, 92.91±2.79%, and 99.16±2.97% respectively. IMB-DG with a mass ratio of IMB to DG of 1:6 was selected for subsequent experiments.

[0093] 1.2 Average particle size, polydispersity index and Zeta potential

[0094] The solid product IMB-DG obtained above (5 mg imatinib and 30 mg dipotassium glycyrrhizinate, with a mass ratio of IMB to DG of 1:6) was dissolved in 10 mL of water, and the average particle size, polydispersity index (abbreviated as PDI), and Zeta potential were measured at 25 °C using a Zetasizer Nano ZS90 (dynamic light scattering method, DLS).

[0095] The results showed that the average particle size of IMB-DG in aqueous solution was 11.26±0.24 nm, the particle size distribution was narrow (PDI = 0.348±0.04), and the Zeta potential was -(21.65±1.02) mV.

[0096] 1.3 Apparent solubility

[0097] Excess IMB and the solid product IMB-DG obtained above (5 mg imatinib and 30 mg dipotassium glycyrrhizinate, with a mass ratio of IMB to DG of 1:6) were separately added to different solvents: water, phosphate buffer solution (PBS), or simulated tear fluid, shaken on a shaker for 24 hours, filtered, and the solubility was measured and calculated.

[0098] The results showed that the apparent solubilities of IMB in water, PBS, and simulated tear fluid were 41.28±7.09, 37.37±1.31, and 61.69±5.55 μg / mL respectively, while the apparent solubilities of IMB in IMB-DG in water, PBS, and simulated tear fluid were 239180.63±68260.89, 333509.93±14837.99, and 359967.62±20059.42 μg / mL respectively.

[0099] Compared with IMB, the solubility of IMB in IMB-DG in water, PBS, and simulated tear fluid was significantly increased.

[0100] Example 2

[0101] Preparation of IMB-DG into eye drops

[0102] The solid product IMB-DG obtained in Example 1 (5 mg imatinib and 30 mg dipotassium glycyrrhizinate, mass ratio of IMB to DG is 1:6) was dissolved in 5 mL of phosphate buffer (PBS: pH = 7.2 - 7.4), and sterilized by filtration through a 0.22 μm filter membrane, obtaining the IMB-DG eye drops (concentration of imatinib is 1 mg / mL).

[0103] Example 3

[0104] IMB-DG was made into an ophthalmic gel

[0105] The solid product IMB-DG obtained in Example 1 (5 mg imatinib and 30 mg dipotassium glycyrrhizinate, mass ratio of IMB to DG is 1:6) was added to 5 mL of phosphate buffer (PBS: pH = 7.2 - 7.4) containing 12.5 mg of hydroxypropyl methylcellulose (HPMC) and 5 mg of sodium hyaluronate (HA), mixed well, and placed at 4 °C overnight, obtaining the IMB-DG gel (content of imatinib is 1 mg / mL).

[0106] Example 4

[0107] Fourier transform infrared spectroscopy (FTIR) and XRD tests

[0108] Fourier transform infrared spectroscopy was used to study the intermolecular interactions in IMB-DG (FTIR, Nicoleti S10, Thermo Fisher, Madison, WI, USA), and X-ray diffraction (XRD, D / max-2400, Rigaku, Japan) was used to study the crystal form changes of IMB in IMB-DG; the results are shown in Figure 2 and Figure 3 respectively. Among them, I.G.M. represents the solid product IMB-DG obtained in Example 1 (5 mg imatinib and 30 mg dipotassium glycyrrhizinate, mass ratio of IMB to DG is 1:6), and I.G.M.H. represents the IMB-DG gel obtained in Example 3.

[0109] The results show that

[0110] (1) In the FTIR spectrum, IMB shows characteristic absorption peaks at 2361 cm -1 , 1650 cm -1 , 1575 cm -1 and 1450 cm -1 . However, the absorption peaks of the IMB part in IMB-DG disappear, and no new peaks appear, indicating that the IMB in IMB-DG is encapsulated by DG, and no chemical reaction occurs during its preparation process;

[0111] (2) In the XRD pattern, IMB has characteristic peaks at 2θ = 5.9°, 14.2°, 17.1° and 24.2°, while these characteristic peaks disappear in the XRD pattern of IMB-DG, indicating that IMB in IMB-DG is amorphous.

[0112] Example 5

[0113] In vitro release test

[0114] Test solution:

[0115] (1) IMB suspension: IMB is dissolved or dispersed in PBS, and the content of IMB is 5 mg / mL;

[0116] (2) The solid product IMB-DG obtained in Example 1 (5 mg imatinib and 30 mg dipotassium glycyrrhizinate, the mass ratio of IMB to DG is 1:6) is dissolved or dispersed in 1 mL of phosphate buffer, and the concentration of IMB is 5 mg / mL;

[0117] (3) The solid product IMB-DG obtained in Example 1 (5 mg imatinib and 30 mg dipotassium glycyrrhizinate, the mass ratio of IMB to DG is 1:6) is added to 1 mL of phosphate buffer containing 12.5 mg of hydroxypropyl methylcellulose (HPMC) and 5 mg of sodium hyaluronate (HA), mixed well, placed at 4 °C overnight, and the concentration of IMB is 5 mg / mL.

[0118] Place 1 mL of the test solution in a dialysis membrane bag (cut-off molecular weight [MWCO] = 14000 Da), then immerse it in 100 mL of PBS (pH = 7.4), culture it on a shaker at 37 °C at 100 rpm, take 1 mL of the dialysis fluid at the set time points (while supplementing 1 mL of PBS), and determine the concentration of IMB by the aforementioned ultraviolet spectrophotometer method, and then calculate the cumulative release amount of IMB. The in vitro release curve is as Figure 4 shown (n = 3).

[0119] The results show that the cumulative release rate of the IMB suspension in 6 hours is only 17.71 ± 0.52%, while the solid products IMB-DG and IMB-DG gel show faster in vitro release rates compared with IMB. Among them, the cumulative release rate of the IMB-DG gel in 6 hours is 53.05 ± 5.95%.

[0120] Example 6

[0121] Eye irritation test

[0122] Male New Zealand white rabbits were randomly divided into the following groups and administered drugs three times a day for one week. At 24 h after the last eye drop, the corneal and conjunctival tissues of the rabbits were observed and recorded using a slit lamp. Then, the corneal and conjunctival tissues were sectioned and stained with hematoxylin-eosin (HE) and observed under a microscope.

[0123] 1) PBS group;

[0124] 2) 0.01 wt.% BAC group: Aqueous solution of benzalkonium chloride (BAC);

[0125] 3) Hyd group: 12.5 mg of hydroxypropyl methylcellulose (HPMC) and 5 mg of sodium hyaluronate (HA) were added to 5 mL of phosphate buffer solution (PBS: pH = 7.2 - 7.4), mixed well, and placed at 4 °C overnight;

[0126] 4) I.G.M. group: The IMB-DG eye drops obtained in Example 2;

[0127] 5) I.G.M.H. group: The IMB-DG gel obtained in Example 3.

[0128] The results were as Figure 5 shown. The 0.01% BAC group showed ocular irritation symptoms such as increased blinking frequency, conjunctival edema, and inflammatory cell infiltration. However, no irritation symptoms such as corneal edema, inflammatory cell infiltration, conjunctival redness, or congestion were observed in the PBS, IMB-DG eye drops (I.G.M. group), and IMB-DG gel (I.G.M.H. group), indicating that PBS, IMB-DG eye drops, and IMB-DG gel do not cause ocular irritation and have good safety.

[0129] Example 7

[0130] 7.1 Establishment of a mouse corneal alkali burn model and grouped drug administration

[0131] After general and local ocular anesthesia, the eyelashes of C57BL / 6J mice were removed, and a filter paper soaked with an alkaline solution (NaOH concentration of 1 M) was placed in the center of the mouse cornea for 30 seconds to induce acute alkali burn of the mouse cornea. The filter paper was removed, and the eyes were rinsed with saline to wash away the residual NaOH solution in the eyes. Alkali burns were performed on both eyes of each mouse.

[0132] After inducing corneal injury, the mice were randomly divided into six groups of 5 mice each, and the following solutions (10 μL each time) were dropped onto the mouse corneas and administered three times a day for 10 days.

[0133] ① PBS group (negative control group);

[0134] ②HA group (positive treatment group): 0.1 wt.% sodium hyaluronate solution;

[0135] ③IMB suspension group: IMB was dissolved or dispersed in PBS, and the content of IMB was 1 mg / mL;

[0136] ④Hyd group: 12.5 mg of hydroxypropyl methylcellulose (HPMC) and 5 mg of sodium hyaluronate (HA) were added to 5 mL of phosphate buffer (PBS: pH = 7.2 - 7.4), mixed well, and placed at 4 °C overnight;

[0137] ⑤I.G.M. group: IMB-DG eye drops obtained in Example 2;

[0138] ⑥I.G.M.H. group: IMB-DG gel obtained in Example 3.

[0139] 7.2 Fluorescein sodium staining of mouse cornea

[0140] On the 1st, 3rd, 5th, 7th, and 10th days, mice in each group were randomly selected. A 0.25% fluorescein sodium solution was applied to the eyes of the mice to cover the eyeballs. After staying on the ocular surface for 10 s, the excess fluorescein sodium solution was rinsed off, the eyeballs were exposed, and observed in front of a slit lamp, and the fluorescein sodium staining of the corneal epithelium of the mice was photographed and recorded. The corneal epithelial staining area was measured using the image analysis software Image J, and the corneal epithelial repair area was calculated.

[0141] The observation results of fluorescein sodium staining are as Figure 6 shown. The results showed that after alkali burn, sodium hydroxide would erode the entire corneal epithelium. After fluorescein sodium staining under cobalt blue light, the corneal epithelium of all groups of mice was fluorescent green (day 0). After administration, IMB-DG eye drops (I.G.M. group) and IMB-DG gel (I.G.M.H. group) were the most effective in treating alkali burns of the mouse cornea and had a therapeutic effect on promoting the healing of epithelial damage.

[0142] According to the corneal epithelial area defect rate (n = 10) statistically analyzed by fluorescein sodium staining, the results are as Figure 7 shown ("*": indicates P < 0.05 compared with the PBS group; "#": indicates P < 0.05 compared with the HA group; "&": indicates P < 0.05 compared with the IMB group; "$": indicates P < 0.05 compared with the Hyd group; "@": indicates P < 0.05 compared with the I.G.M. group).

[0143] The results showed that compared with the PBS group (negative control), IMB-DG eye drops (I.G.M. group) and IMB-DG gel (I.G.M.H. group) could effectively repair the corneal epithelial damage caused by alkali burn and accelerate the healing of corneal injury.

[0144] In particular, the corneal injury healing rates on the 5th and 10th days of treatment with IMB-DG gel (corneal injury healing rate = 1 - corneal epithelial area defect rate) were 93.25 ± 3.19% and 99.25 ± 0.83% respectively, which were superior to the corneal injury healing rates on the 5th and 10th days of the PBS group (46.23 ± 2.58%, 76.78 ± 2.11%) and the corneal injury healing rates on the 5th and 10th days of the HA group (75.53 ± 1.80%, 92.73 ± 1.08%).

[0145] 7.3 Corneal opacity evaluation

[0146] Using a slit lamp microscope to observe, the corneal opacity was evaluated and scored (n = 10): the corneal opacity was graded from 0 to 5, with the highest score of 5 points, and the scoring results were as Figure 8 shown ("*": indicates P < 0.05 compared with the PBS group; "#": indicates P < 0.05 compared with the HA group; "&": indicates P < 0.05 compared with the IMB group; "$": indicates P < 0.05 compared with the Hyd group; "@": indicates P < 0.05 compared with the I.G.M. group).

[0147] 7.4 Corneal sensitivity test

[0148] Detected using a Cochet-Bonnet corneal esthesiometer (n = 10), and the results were as Figure 9 shown ("*": indicates P < 0.05 compared with the PBS group; "#": indicates P < 0.05 compared with the HA group; "&": indicates P < 0.05 compared with the IMB group; "$": indicates P < 0.05 compared with the Hyd group; "@": indicates P < 0.05 compared with the I.G.M. group).

[0149] The results showed that alkali burn severely damaged the corneal sensitivity, and the corneal sensitivity in the PBS group recovered slowly; compared with the PBS group, the corneal sensitivity of IMB-DG eye drops (I.G.M. group) and IMB-DG gel (I.G.M.H. group) was significantly improved.

[0150] 7.5 Observation of corneal neovascularization

[0151] Using a slit lamp microscope to observe, the corneal neovascularization was evaluated and scored (n = 10): the score was between 0 and 4, and the results were as Figure 10 and Figure 11Shown as follows (“*”: indicates P < 0.05 compared with the PBS group; “#”: indicates P < 0.05 compared with the HA group; “&”: indicates P < 0.05 compared with the IMB group; “$”: indicates P < 0.05 compared with the Hyd group; “@”: indicates P < 0.05 compared with the I.G.M. group).

[0152] The results showed that compared with the PBS group, the IMB-DG eye drops (I.G.M. group) and the IMB-DG gel (I.G.M.H. group) had a significant inhibitory effect on the neovascularization caused by corneal alkali burn. In particular, the effect of the IMB-DG gel was the best.

[0153] 7.6 Histopathological examination

[0154] On the 10th day, mice in each group were randomly selected and sacrificed by cervical dislocation. The eyeballs were removed for histopathological examination.

[0155] The results showed that the corneas of the PBS and IMB groups were still damaged, showing symptoms such as corneal epithelial defect and inflammatory cell infiltration, while the tissue structure of the IMB-DG gel (I.G.M.H. group) was normal and no obvious inflammatory cell infiltration was seen.

[0156] Of course, the present invention can also have many other embodiments. Without departing from the spirit and essence of the present invention, those skilled in the art can make various corresponding changes and / or deformations according to the present invention, and these corresponding changes and / or deformations should all fall within the protection scope of the appended claims of the present invention.

Claims

1. Use of a piperazine compound or a composition thereof in the preparation of an ophthalmic drug for preventing and / or treating alkali burns, characterized in that, The described composition contains piperazine compounds and glycyrrhizinate; the piperazine compounds are imatinib or its pharmaceutically acceptable salts; the glycyrrhizinate is selected from one or more of sodium glycyrrhizinate, disodium glycyrrhizinate, potassium glycyrrhizinate, dipotassium glycyrrhizinate, ammonium glycyrrhizinate, and diammonium glycyrrhizinate; Preferably, the piperazine compound is imatinib; and / or, the glycyrrhizinate is dipotassium glycyrrhizinate or disodium glycyrrhizinate.

2. The use according to claim 1, characterized in that, The solvent of the piperazine compound or its composition is water or phosphate buffer solution, and the ophthalmic drug is eye drops.

3. The use according to claim 1, characterized in that, The solvent of the piperazine compound or its composition is water or phosphate buffer solution, the composition contains cellulose derivatives and hyaluronic acid compounds, and the ophthalmic drug is an ophthalmic gel; Among them, the cellulose derivatives are selected from one or more of methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, methyl ethyl cellulose, hydroxyethyl cellulose, and hydroxyethyl methyl cellulose; the hyaluronic acid compounds are hyaluronic acid or its salts; Preferably, the cellulose derivative is hydroxypropyl methyl cellulose; the hyaluronic acid compound is an alkali metal salt of hyaluronic acid, more preferably sodium hyaluronate.

4. A pharmaceutical composition, characterized in that, Containing piperazine compounds and pharmaceutically acceptable excipients; the piperazine compounds are imatinib or its pharmaceutically acceptable salts, and the pharmaceutically acceptable excipients contain glycyrrhizinate; Preferably, the mass ratio of the piperazine compound to the glycyrrhizinate is 1:1 to 20; and / or, the glycyrrhizinate is selected from one or more of sodium glycyrrhizinate, disodium glycyrrhizinate, potassium glycyrrhizinate, dipotassium glycyrrhizinate, ammonium glycyrrhizinate, and diammonium glycyrrhizinate; and / or, the piperazine compound is imatinib; More preferably, the mass ratio of the piperazine compound to the glycyrrhizinate is 1:4 to 10, and further preferably the mass ratio of the piperazine compound to the glycyrrhizinate is 1:5.5 to 6.5; and / or, the glycyrrhizinate is dipotassium glycyrrhizinate or disodium glycyrrhizinate.

5. The pharmaceutical composition according to claim 4, wherein The piperazine compound exists in the pharmaceutical composition in an amorphous form; and / or, the encapsulation efficiency of the piperazine compound is at least 80%; preferably, the encapsulation efficiency of the piperazine compound ≥ 90% or ≥ 95%.

6. The pharmaceutical composition according to claim 4, wherein The pharmaceutical composition is a liquid preparation, semi-solid preparation or solid preparation; and / or, the piperazine compound in the pharmaceutical composition is in a therapeutically effective amount.

7. The pharmaceutical composition according to any one of claims 4 to 6, characterized in that, The pharmaceutical composition is a liquid preparation, the solvent of the pharmaceutical composition is water or phosphate buffer solution, and the liquid preparation is eye drops; Preferably, when the concentration of the piperazine compound in the liquid preparation is 0.5 mg / mL, the liquid preparation meets one or more of the following conditions ① to ③: ①. The average particle size of the liquid preparation is 1 to 100 nm, preferably 5 to 20 nm; ②. The polydispersity coefficient of the liquid preparation is ≤ 0.6, preferably 0.1 to 0.5; ③ The Zeta potential of the liquid preparation is -30 to -1 mV, preferably -25 to -15 mV; More preferably, the liquid preparation simultaneously satisfies the above conditions ① to ③.

8. The pharmaceutical composition according to any one of claims 4 to 6, characterized in that, The pharmaceutical composition is a semi-solid preparation, which contains a cellulose derivative and a hyaluronic acid compound. The solvent of the pharmaceutical composition is water or a phosphate buffer solution, and the semi-solid preparation is an ophthalmic gel; Preferably, The mass ratio of the piperazine compound to the cellulose derivative is 1:1 to 20; And / or, the mass ratio of the piperazine compound to the hyaluronic acid compound is 1:0.5 to 20; And / or, the cellulose derivative is selected from one or more of methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, calcium carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylcellulose, methylethylcellulose, hydroxyethylcellulose, and hydroxyethylmethylcellulose; And / or, the hyaluronic acid compound is hyaluronic acid or its salt; More preferably, The mass ratio of the piperazine compound to the cellulose derivative is 1:2 to 5, and further preferably 1:2.5; And / or, the mass ratio of the piperazine compound to the hyaluronic acid compound is 1:0.9 to 1.5, and further preferably 1:1; And / or, the cellulose derivative is hydroxypropylmethylcellulose; And / or, the hyaluronic acid compound is an alkali metal salt of hyaluronic acid, and further preferably sodium hyaluronate.

9. The pharmaceutical composition according to any one of claims 4 to 6, characterized in that, The pharmaceutical composition is a pharmaceutical composition for preventing and / or treating ocular chemical burns; Preferably, the ocular chemical burn is an alkali burn.

10. A method for preparing the pharmaceutical composition according to any one of claims 4 to 9, characterized in that, Comprising the following steps: Dissolving or dispersing the piperazine compound and the glycyrrhizinate in an organic solvent, and then removing the organic solvent to obtain a product; Optionally, a formulation step may be included: dissolving or dispersing the aforementioned product in the solvent of the pharmaceutical composition, or dissolving or dispersing the aforementioned product together with the cellulose derivative and the hyaluronic acid compound in the solvent of the pharmaceutical composition, thus obtaining the product; Preferably, the organic solvent is an alcohol solvent; and / or, the amount of the organic solvent corresponding to each milligram of the piperazine compound is 0.1 to 20 mL; More preferably, the organic solvent is ethanol; and / or, the amount of the organic solvent corresponding to each milligram of the piperazine compound is 1 to 10 mL.

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