Formula of eye drops with solubilizing and permeation promoting effects

Abstract

The invention provides application of a medicinal auxiliary material diethylene glycol monoethyl ether as an eye drop auxiliary material, and belongs to the technical field of medicinal preparations. 0.01%-20% diethylene glycol monoethyl ether normal saline solution is good in stability, free of irritation to eyes and capable of achieving the transcorneal penetration promoting effect, medicine solubility is improved, meanwhile, medicine absorption and permeation are promoted, and the eye drop effect is improved. The method has potential medical value and commercial value in the aspects of preparation and application of the eye drops, especially the eye drops for reducing intraocular pressure.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations and relates to an eye drop formula with solubilizing and penetrating-promoting effects. Background Art

[0002] Conventional eye drops are based on water for injection as solvent, or a small amount of surfactant (tween80 / 20, CMC-Na, POLOXAMER 188 / 407, glyceryl monostearate, HEC, hydroxypropyl gamma-cyclodextrin, OP-40, PEG fatty acid esters, etc.) is added to the aqueous solution to improve the solubility of poorly soluble drugs, thereby meeting the requirements of eye drop formulations. Penetration-enhancing technology is mostly used in the field of transdermal administration or cosmetics. In addition to pharmaceutical technology, physical penetration-enhancing technology, and biological penetration-enhancing technology, commonly used chemical penetration enhancers include ethanol, propylene glycol, azone, organic acids, and surfactants. Traditional Chinese medicine penetration enhancers include borneol, eucalyptus, and mint.

[0003] However, when encountering extremely insoluble drugs, the existing injection water formula and surfactants cannot meet the needs of solubilization and dissolution-aiding, and the penetration-promoting effect is poor. It is necessary to explore new compound solvent eye drop formulas.

[0004] Diethylene glycol monoethyl ether is a protic solvent with the appearance of a colorless and transparent solution. It is a strong solubilizer that can increase the solubility of lipophilic and hydrophilic drugs. In topical preparations, diethylene glycol monoethyl ether is a highly effective penetration enhancer with a clear mechanism of effective penetration enhancement: reducing drug charge through solvent effect, increasing drug solubility and distribution in the stratum corneum, and maintaining hydration dynamics in the stratum corneum without disrupting the lipid bilayer structure. It has toxicology-related studies and a history of use in approved drugs, with high safety and low irritation.

[0005]

[0006] Diethylene glycol monoethyl ether (Molecular formula C6H 14 O3, molecular weight 134.17) Summary of the invention

[0007] The purpose of the present invention is to solve the problem of drug delivery of existing poorly soluble drugs to the eye, especially to the posterior chamber, by using commercial pharmaceutical excipients and simple preparation processes. The present invention develops an eye drop formula containing: diethylene glycol monoethyl ether, a pH regulator, an osmotic pressure regulator and a solvent, which solves the problem of promoting penetration while solubilizing the drug.

[0008] The present invention is achieved through the following technical solutions: Firstly, the invention provides an eye drop formula with solubilizing and penetration-promoting effects, which contains at least diethylene glycol monoethyl ether.

[0009] Secondly, in the formula of the present invention, the preferred safe concentration ratio of diethylene glycol monoethyl ether is 0.01%-20%.

[0010] Again, for solubilization, the preferred concentration ratio of diethylene glycol monoethyl ether is 0.01%-20%.

[0011] Furthermore, poorly soluble drugs solubilized by the present invention include but are not limited to: cilostazol, cyclosporine, sirolimus, tacrolimus, latanoprost, and difluprednate.

[0012] Again, in order to achieve drug delivery to the posterior chamber of the eye, the preferred concentration ratio of diethylene glycol monoethyl ether is 5%-20%.

[0013] The beneficial effects of the present invention are as follows: according to the safety evaluation of the eye irritation and other safety evaluations of the formula and the MRI verification of transcorneal delivery, a 0.01%-20% diethylene glycol monoethyl ether physiological saline solution can be used as an eye drop solubility aid and penetration enhancer, thereby increasing the solubility of the drug and achieving transcorneal delivery, being suitable for solubilization and penetration enhancement of intraocular pressure-lowering drugs and improving the efficacy of the intraocular pressure-lowering drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 . T1 sequence time-imaging changes Figure 2 . T2 sequence time-imaging changes Implementation

[0015] The present invention discloses an eye drop formula with solubilizing and permeation-promoting effects, which contains at least diethylene glycol monoethyl ether, and studies the safety and hyperosmotic application in ophthalmic preparations. Those skilled in the art can refer to the content of the present invention, and all similar substitutions and modifications are obviously considered to be included in the scope of the present invention. The application of the present invention has been described through better embodiments, and relevant personnel can obviously modify or appropriately change the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0016] The present invention is further described below by way of examples, but the examples do not limit the present invention in any way. Example 1

[0017] Effect of diethylene glycol monoethyl ether on hemolysis of erythrocytes in rats 1 Materials and methods 15-20 mL of blood was collected from rats (Sprague-Dawley (SD) rats, Shanghai Slake Laboratory Animal Co., Ltd.) through the abdominal aorta and placed in an appropriate container; a stirrer (or glass beads) was placed in the container and stirred slowly for several minutes to remove fibrinogen from the blood and make it defibrinated blood; about 10 times the amount of 0.9% sodium chloride injection was added, shaken and dispensed into centrifuge tubes, centrifuged at 1500r / min for 15 minutes, and the supernatant was removed; the precipitated red blood cells were washed with 0.9% sodium chloride injection according to the above method until the supernatant was no longer red; the red blood cells were prepared with 0.9% sodium chloride injection to make a 2% red blood cell suspension. Different concentrations of diethylene glycol monoethyl ether (purchased from Gattefossé, France) solution (5%-40%) were mixed with 2% red blood cell suspension in a volume ratio of 1:1, and the total incubation volume was 500ul. Incubated at 32℃ for 10min, and centrifuged at 10000rpm for 1min to terminate the reaction. 200ul of supernatant was aspirated and placed in a 96-well plate. The absorbance was measured at 540nm using a microplate reader. Physiological saline was used as a control for autohemolysis, and 100mg / L SDS was used as a control for total hemolysis. Three parallel wells were measured in each group.

[0018] The hemolysis rate calculation formula of sample solutions with different concentrations is: Hemolysis rate % (540nm) = (sample absorption - negative control absorption) / (full hemolysis control absorption - negative control absorption) × 100 2 Experimental Results A hemolysis rate of more than 5% is considered hemolysis. Experimental observations show that the supernatant of the negative control group is clear and transparent, while the supernatant of the full hemolysis control group is red. Compared with the negative control group, when the concentration of diethylene glycol monoethyl ether (HTP) is ≥20%, the hemolysis rate exceeds 5%, and the color of the supernatant changes, indicating that red blood cell hemolysis has occurred.

[0019] Table 1. Hemolytic effect of different concentrations of diethylene glycol monoethyl ether on rat erythrocytes Group (final concentration) Hemolysis rate% Negative control group 0.00±0.00 5% diethylene glycol monoethyl ether -2.7±0.0005 10% Diethylene glycol monoethyl ether -2.6±0.0009 20% Diethylene glycol monoethyl ether 0.5±0.0005 25% Diethylene glycol monoethyl ether 9.0±0.0003 30% Diethylene glycol monoethyl ether 43.9±0.002 35% Diethylene glycol monoethyl ether 52.0±0.007 40% Diethylene glycol monoethyl ether 65±0.014 100mg / L SDS (100 μM) 100.0±0.00 Data are expressed as Mean±SD. Example 2

[0020] Effects of diethylene glycol monoethyl ether on the viability of human corneal epithelial cells 1 Materials and methods Human corneal epithelial cell line (HCE-T cells, purchased from Pronocell Bio) was cultured in an incubator at 37°C, 5% carbon dioxide, and 70%-80% humidity. When the cell density reached 80%-90%, the cells were plated for experiments. The specific operation is as follows: discard the culture supernatant and rinse the cells 1-2 times with PBS without calcium and magnesium ions. Add 0.25% (w / v) trypsin-0.53 mM EDTA to the culture bottle (1-2mL for T25 bottle, 2-3mL for T75 bottle), place it in a 37°C incubator for digestion for 1-2 minutes, and then observe the cell digestion under a microscope. If most of the cells become round and fall off, quickly take it back to the operating table, tap the culture bottle a few times, and add 3-4ml of culture medium containing 10% FBS to terminate the digestion. Gently mix and aspirate, centrifuge at 1000rpm for 3-5min, discard the supernatant, add 1-2mL of culture medium and blow it evenly. The cell suspension was plated in a 96-well plate at an appropriate density. After the cells adhered to the wall, different concentrations of diethylene glycol monoethyl ether were added, and the final concentration was set to 0.16% (Control), 0.31%, 0.63%, 1.25%, 2.5%, 5%, 10%, 15%, 20%, and 4 replicates were set for each concentration. 30 minutes after administration, the cell viability was measured using the Cell Titer-Glo luminescent live cell assay to evaluate the irritation of different concentrations of diethylene glycol monoethyl ether to corneal epithelial cells.

[0021] 2 Experimental Results The experimental results were analyzed by one-way ANOVA and showed that compared with the control group, the cell viability of the 15% and 20% diethylene glycol monoethyl ether groups was significantly reduced, indicating that it had a significant stimulating effect on human corneal epithelial cells. Concentrations of 10% and below had no significant stimulating effect on corneal epithelial cells and had good safety.

[0022] Table 2. Effects of different concentrations of diethylene glycol monoethyl ether on the viability of human corneal epithelial cells Group (final concentration) Cell viability% Control Group 100.00±18.54 0.31%Diethylene glycol monoethyl ether 105.53±15.97 0.63%Diethylene glycol monoethyl ether 106.26±14.83 1.25% Diethylene glycol monoethyl ether 108.65±14.49 2.5% diethylene glycol monoethyl ether 108.78±9.61 5% diethylene glycol monoethyl ether 94.96±11.51 10% Diethylene glycol monoethyl ether 94.21±13.25 15% Diethylene glycol monoethyl ether 60.20±7.49** 20% Diethylene glycol monoethyl ether 12.76±2.61**** Data are expressed as Mean ± SD. **p < 0.01, ****p < 0.0001 compared with Control Example 3

[0023] Penetration effect of 10% diethylene glycol monoethyl ether in eyes 1 Materials and methods Isoflurane (purchased from Rayward), Rayward small animal anesthesia system, mice (C57BL / 6 mice, Shanghai Slake Laboratory Animal Co., Ltd.), Bruker BioSpec 94 / 20 USR 9.4 T small animal magnetic resonance imaging instrument (Animal Imaging Research Center, Nanjing Drum Tower Hospital).

[0024] After the mouse was anesthetized with isoflurane, it was placed on the transmission track of the small animal magnetic resonance imaging instrument. The head of the mouse was fixed in a prone position using a mouse head volume coil with an inner diameter of 23 mm, and the body was straightened. Since the solvent to be tested (10% diethylene glycol monoethyl ether) is an aqueous solution, there is no need to add an MRI probe, because water increases the signal intensity of MRI T2 and allows the solution molecules to be visualized. A pre-scan image was obtained by rapid scanning with the simplest sequence. According to the pre-scan image, the animal positioning and magnetic field uniformity were adjusted. The eye scanning position was enlarged, and an additional magnetic field was added to make the scanning site and other areas have a magnetic field difference (i.e., gradient difference). Radio frequency signals were transmitted to the scanning area, and the coil received analog signals to read clear eye images. After the T1 / T2 sequence obtained the physiological baseline image, two drops of 10% diethylene glycol monoethyl ether saline solution were added to the left eye of the mouse, and an equal volume of saline was given to the right eye. The T1 / T2 sequence alternately acquired images every 10-15 minutes.

[0025] 2 Experimental Results MRI obtains images of the entire head, such as Figure 1 , Figure 2 , 16 coronal planes were evenly divided anteriorly and posteriorly, and it was observed that the largest range of ocular vision could be obtained on the 7th coronal plane. The observable ocular structures included the anterior chamber, lens, vitreous body and other tissue structures around the eye. The images were collected continuously for 90 min. The T1 sequence observed that compared with the time-base image without drug administration, the left eye with 10% diethylene glycol monoethyl ether showed that the solution was evenly spread on the cornea, and the brightness was higher than that of the right eye in the saline group. The T2 sequence showed that compared with the saline group, 10% diethylene glycol monoethyl ether could penetrate the cornea to reach the ciliary body, and showed a high-brightness mode within the 1.5h observation time range, indicating that the solution could stay in the ciliary body for a long time. This shows that diethylene glycol monoethyl ether has the potential to carry drugs across the corneal barrier to the ciliary body, which is helpful for the delivery of eye drops that need to cross the corneal barrier to work, such as intraocular pressure-lowering drugs.

Claims

1. An eye drop formula with solubilizing and penetration-promoting effects, the formula contains diethylene glycol monoethyl ether, and the diethylene glycol monoethyl ether is used as a cosolvent and a penetration-promoting agent.

2. The formulation of claim 1, Features The preferred safe concentration ratio of diethylene glycol monoethyl ether is 0.01-20%, and the remainder can be used as a solvent, pH adjuster, and osmotic pressure regulator for eye drops.

3. The formulation of claim 2, Features The preferred solubilization concentration ratio of diethylene glycol monoethyl ether is 0.01%-20%.

4. The formulation of claim 1, Features The poorly soluble drugs solubilized by the present invention include but are not limited to: cyclosporine, sirolimus, tacrolimus, latanoprost, and difluprednate.

5. The formulation of claim 1, Features In order to achieve drug delivery to the posterior chamber of the eye, the preferred concentration ratio of diethylene glycol monoethyl ether is 5%-20%.

6. The formulation of claim 2, wherein the remainder is a solvent for eye drops Features: Purified water, water for injection, purified water containing sodium carboxymethyl cellulose, purified water for injection containing sodium carboxymethyl cellulose.

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