Long-acting sustained release preparation for preventing or treating retinal damage, and preparation method thereof

Abstract

The invention belongs to the field of pharmaceutical preparations, and relates to a long-acting sustained release preparation for preventing or treating retinal damage, and a preparation method thereof. The long-acting sustained release preparation takes erythropoietin (EPO) as an active component, takes dextran as a protective agent for the active component, and takes poly(lactic-co-glycolic acid), polylactic acid or polycaprolactone as a coating component to prepare sustained release microspheres, wherein the erythropoietin is coated by the poly(lactic-co-glycolic acid), polylactic acid or polycaprolactone. Proven by animal experiments, the sustained release microspheres of the long-acting sustained release preparation provided by the invention have the same protective actions on the ganglionic cells of damaged retinas by single vitreous chamber injection and repeated EPO protein vitreous chamber injection, and the sustained release microspheres are capable of avoiding a series of complications caused by many times of injection and overcoming the defects of repeated intraocular injection administration and gene therapy. The long-acting sustained release preparation provided by the invention adopts intraocular local administration, can reduce the dosage and treatment cost, and can not generate adverse effects on other organs or tissues in vivo.

Description

technical field

[0001] The invention belongs to the field of pharmaceutical preparations, and relates to a long-acting slow-release preparation for preventing or treating retinal damage and a preparation method thereof. Background technique

[0002] It has been reported in the literature that there are various types of retinal optic nerve diseases, which are one of the important blinding factors. At present, there is no effective treatment method in ophthalmology clinic. Studies have shown that after the optic nerve is injured, the axon of retinal ganglion cells is damaged, resulting in the blockage of axoplasmic transport at the cribriform plate of the axons, resulting in insufficient supply of neurotrophic factors, release of excitatory neurotransmitters, oxidative stress and A large number of free radicals are produced, affecting the normal function of retinal ganglion cells (RGCs), causing their degeneration and death; the toxic media produced by the death of retinal gan...

Examples

Embodiment 1

[0052] ①Preparation of erythropoietin-dextran microparticles

[0053] a) Pre-preparing erythropoietin with a weight percentage of 0.5%, 10%, or 20% of the erythropoietin microspheres into a solution with a weight percentage concentration of 0.01%, 10%, or 20%, respectively, Dextran (with a molecular weight of 10,000, 2,500,000, or 5,000,000 Daltons) accounted for 0.5%, 15%, or 30% by weight of the erythropoietin microspheres was formulated to have a concentration of 1%, 15% by weight, respectively. % or 30% solution; polyethylene glycol (molecular weight: 2,000, 150,000, or 300,000) is prepared into a solution with a concentration of 1%, 20%, or 40% by weight respectively.

[0054] b) Mix the above erythropoietin solution according to the volume ratio of erythropoietin solution: dextran solution: PEG solution = 50:1:200, 1:1:8 or 2:3:12 and mix well ;

[0055] c) Then mix the above step b) evenly, pre-freeze in the freezer for 8-32 hours, and then freeze-dry,

[0056] d) Di...

Embodiment 2

[0064] ①Preparation of erythropoietin-dextran microparticles

[0065] With step 1. of embodiment 1.

[0066] a) Take 1 mg, 2.5 mg or 3 mg of the erythropoietin microparticles prepared in the corresponding proportion of step ① and add them to 330 mg of polylactic acid-glycolic acid (PLGA), polylactic acid (PLA) with a concentration of 30% by weight. , or a dichloromethane solution of polycaprolactone (PCL) (the prepared erythropoietin sustained-release microspheres contain 0.5% erythropoietin and 0.5% dextran), and the weight percent concentration is 15% 50mg of polylactic acid-glycolic acid (PLGA), polylactic acid (PLA), or polycaprolactone (PCL) in dichloromethane solution (the prepared erythropoietin sustained-release microspheres contain 10% erythropoietin and 15% dextran), or 5% of 140mg polylactic-glycolic acid (PLGA), or polycaprolactone (PCL) in dichloromethane solution (the prepared erythropoietin sustained-release microspheres containing 20% erythropoietin and 30% d...

Embodiment 3

[0072] Apoptosis in situ cell apoptosis detection method (TUNEL detection) to observe the protective effect of the long-acting sustained-release preparation EPO sustained-release microspheres of the present invention on damaged retinal ganglion cells (RGCs)

[0073] ①Experimental animals and grouping: Adult male SD rats (about 200g), after 7 days of adaptive feeding, check both eyes, check that the refractive interstitium is clear, the pupils are equal in size and round, sensitive to light reflex, and the fundus is normal. The left eye was regarded as the optic nerve contusion eye, and the right eye was regarded as the normal control eye. According to the intravitreal injection administration after optic nerve contusion, they were divided into the following five groups: untreated group (no injection into vitreous cavity), EPO group (injection of EPO into vitreous cavity), EPO-PLGA group (injection of EPO-PLGA microspheres into vitreous cavity) , PBS group (intravitreal injecti...

technical field

[0001] The invention belongs to the field of pharmaceutical preparations, and relates to a long-acting slow-release preparation for preventing or treating retinal damage and a preparation method thereof. Background technique

[0002] It has been reported in the literature that there are various types of retinal optic nerve diseases, which are one of the important blinding factors. At present, there is no effective treatment method in ophthalmology clinic. Studies have shown that after the optic nerve is injured, the axon of retinal ganglion cells is damaged, resulting in the blockage of axoplasmic transport at the cribriform plate of the axons, resulting in insufficient supply of neurotrophic factors, release of excitatory neurotransmitters, oxidative stress and A large number of free radicals are produced, affecting the normal function of retinal ganglion cells (RGCs), causing their degeneration and death; the toxic media produced by the death of retinal gan...

Examples

Embodiment 1

[0052] ①Preparation of erythropoietin-dextran microparticles

[0053] a) Pre-preparing erythropoietin with a weight percentage of 0.5%, 10%, or 20% of the erythropoietin microspheres into a solution with a weight percentage concentration of 0.01%, 10%, or 20%, respectively, Dextran (with a molecular weight of 10,000, 2,500,000, or 5,000,000 Daltons) accounted for 0.5%, 15%, or 30% by weight of the erythropoietin microspheres was formulated to have a concentration of 1%, 15% by weight, respectively. % or 30% solution; polyethylene glycol (molecular weight: 2,000, 150,000, or 300,000) is prepared into a solution with a concentration of 1%, 20%, or 40% by weight respectively.

[0054] b) Mix the above erythropoietin solution according to the volume ratio of erythropoietin solution: dextran solution: PEG solution = 50:1:200, 1:1:8 or 2:3:12 and mix well ;

[0055] c) Then mix the above step b) evenly, pre-freeze in the freezer for 8-32 hours, and then freeze-dry,

[0056] d) Di...

Embodiment 2

[0064] ①Preparation of erythropoietin-dextran microparticles

[0065] With step 1. of embodiment 1.

[0066] a) Take 1 mg, 2.5 mg or 3 mg of the erythropoietin microparticles prepared in the corresponding proportion of step ① and add them to 330 mg of polylactic acid-glycolic acid (PLGA), polylactic acid (PLA) with a concentration of 30% by weight. , or a dichloromethane solution of polycaprolactone (PCL) (the prepared erythropoietin sustained-release microspheres contain 0.5% erythropoietin and 0.5% dextran), and the weight percent concentration is 15% 50mg of polylactic acid-glycolic acid (PLGA), polylactic acid (PLA), or polycaprolactone (PCL) in dichloromethane solution (the prepared erythropoietin sustained-release microspheres contain 10% erythropoietin and 15% dextran), or 5% of 140mg polylactic-glycolic acid (PLGA), or polycaprolactone (PCL) in dichloromethane solution (the prepared erythropoietin sustained-release microspheres containing 20% erythropoietin and 30% d...

Embodiment 3

[0072] Apoptosis in situ cell apoptosis detection method (TUNEL detection) to observe the protective effect of the long-acting sustained-release preparation EPO sustained-release microspheres of the present invention on damaged retinal ganglion cells (RGCs)

[0073] ①Experimental animals and grouping: Adult male SD rats (about 200g), after 7 days of adaptive feeding, check both eyes, check that the refractive interstitium is clear, the pupils are equal in size and round, sensitive to light reflex, and the fundus is normal. The left eye was regarded as the optic nerve contusion eye, and the right eye was regarded as the normal control eye. According to the intravitreal injection administration after optic nerve contusion, they were divided into the following five groups: untreated group (no injection into vitreous cavity), EPO group (injection of EPO into vitreous cavity), EPO-PLGA group (injection of EPO-PLGA microspheres into vitreous cavity) , PBS group (intravitreal injecti...

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