CN107137698A - A kind of being used for comprising GDNF treats the pharmaceutical composition of corneal epithelial wound - Google Patents

Abstract

The invention discloses a kind of being used for comprising GDNF treat the pharmaceutical composition of corneal epithelial wound, described pharmaceutical composition contains GDNF (preferably people source), and preferably also containing OPG (preferably people source).The pharmaceutical composition (being preferably the form of eye drops) of the present invention can not only promote limbal stem cell to breed and clonality, the injury repair of corneal epithelium can also be effectively facilitated, includes reparation, the treatment of diabetes keratopathy, the treatment of nerve keratitis, the treatment of limbal stem cell deficiencies of persistent corneal epithelial damage.

Description

A pharmaceutical composition containing GDNF for treating corneal epithelial damage

technical field

The invention relates to the technical field of medicine, in particular to a pharmaceutical composition for treating corneal epithelial damage (preferably corneal epithelial damage induced by diabetes).

Background technique

The transparency and functional integrity of the cornea depend on the normal structure and metabolism of each layer of the cornea, and the outermost corneal epithelial tissue undoubtedly plays a very important role in this. Various physical damage, chemical damage, mechanical damage, pathogenic microorganisms, endocrine and immune factors can cause corneal epithelial damage. The corneal epithelium is a physical barrier against the invasion of external pathogenic factors. The maintenance of its integrity depends on the tight junction and anchor junction between epithelial cells, cells and basement membrane, and the continuous self-renewal of epithelial cells. Damage to the corneal epithelium can affect the connection between cells, causing changes in the permeability and selectivity of the cell membrane, thereby affecting its barrier function, making the cornea vulnerable to external pathogenic factors, causing inflammation, corneal opacity, and even blindness. Persistent corneal epithelial injury can cause pathogenic microorganisms to easily pass through the Bowman's membrane and invade the corneal parenchyma due to the destruction of the normal and complete epithelial barrier, causing corneal inflammation, corneal stroma turbidity, and collagen melting to form corneal ulcers, corneal perforation, and even Eyeball loss with serious consequences.

At present, the traditional treatment methods for persistent corneal epithelial injury include: tear substitutes, contact lenses, eyelid suture, growth factors, autologous serum, etc., but the treatment effect is not good.

Glial cell-derived neurotrophic factor (GDNF) is an important neurotrophic factor. Patent document 96198645.X discloses a method for treating retinal ganglion cell damage (especially glaucoma-related optic nerve damage) with GDNF protein product. OPG (osteoprotegerin) is commonly used to treat osteoporosis. But there is no report that the combination of GDNF and OPG is specially used for repairing corneal epithelial damage.

Contents of the invention

The invention provides a pharmaceutical composition for treating corneal epithelial injury, the pharmaceutical composition includes GDNF and OPG.

Preferably, the content of GDNF in the pharmaceutical composition is 0.001-1wt%;

Preferably, the content of OPG in the pharmaceutical composition is 0.001-0.1wt%;

Preferably, the GDNF is mouse GDNF or human GDNF, more preferably human GDNF;

In a preferred embodiment of the present invention, the human GDNF is human recombinant GDNF.

Preferably, the OPG is murine OPG or human OPG, more preferably human OPG;

In a preferred embodiment of the present invention, the human OPG is human recombinant OPG.

Preferably, the above-mentioned pharmaceutical composition can be prepared into various preparations for topical administration, gastrointestinal administration or parenteral administration; the topical administration preparation is a preparation for ophthalmic administration, such as: Eye drops, injections, powder injections, eye ointments, emulsions, liposomes, microcapsules, gels, implants, inserts, ophthalmic films, inclusion eye drops and others can be used for ocular administration The formulation for parenteral administration is suitable for intravenous injection, intramuscular injection, subcutaneous injection, bone marrow injection, transdermal administration, mucosal administration, inhalation administration and other dosage forms;

In a preferred embodiment of the present invention, the pharmaceutical composition is a preparation for ophthalmic administration, preferably selected from: eye drops, eye ointment, gel and ophthalmic film.

When the pharmaceutical composition of the present invention for treating dry eye is an ophthalmic solution, it is provided in any dosage form for an ophthalmic solution, for example, an aqueous eye drop (such as an aqueous ophthalmic solution, aqueous suspension ophthalmic solution, viscous ophthalmic solution, solubilized ophthalmic solution, etc.), or a nonaqueous ophthalmic solution (such as nonaqueous ophthalmic solution and nonaqueous ophthalmic solution suspension).

Preferably, the pharmaceutical composition is an aqueous eye drop, wherein the content of GDNF is 1 ng/ml-1000 μg/ml, preferably 10-5000 ng/ml, more preferably 100-1000 ng/ml; and/or, The content of OPG is 1ng/ml-100ng/ml, preferably 10-50ng/ml;

Preferably, the above pharmaceutical composition also includes pharmaceutically acceptable additives;

Preferably, the additive is selected from one or more of: bacteriostats, pH regulators, osmotic pressure regulators, solubilizers (stabilizers), thickeners, chelating agents, etc.;

Preferably, the bacteriostatic agent includes, but is not limited to: one or more combinations of thimerosal, quaternary ammonium salts, dumiphene, chlorhexidine, chlorobutanol, parabens, and sorbic acid; Preferably, the quaternary ammonium salt antibacterial agent such as benzalkonium chloride, benzalkonium bromide, parabens such as ethylparaben, etc.;

Preferably, the preservative content in the pharmaceutical composition is 0.001-1.0%;

Preferably, the pH regulator includes, but is not limited to: one of phosphate, acetate, citric acid and its salts, carbonate solution, sodium hydroxide, potassium hydroxide, hydrochloric acid, boric acid, phosphoric acid, etc. or more;

Preferably, the pH in the pharmaceutical composition is 5.5-7.5;

Preferably, the osmotic pressure regulator includes but not limited to: sugars (such as sorbitol, glucose, mannitol), polyalcohols (such as glycerin, polyethylene glycol, polypropylene glycol), salts such as sodium chloride ;

Preferably, the solubilizer includes, but is not limited to: cyclodextrin and its derivatives, water-soluble polymers (such as polyvinylpyrrolidone), surfactants (such as polysorbate 80, trade name: Tween 80) ;

Preferably, the thickener is selected from one or more of methylcellulose, carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose and salts thereof kind;

Preferably, the chelating agent includes, but is not limited to: sodium edetate, sodium citrate, and sodium condensed phosphate.

In one embodiment of the present invention, the pharmaceutical composition is ophthalmic gel, ophthalmic ointment, which also includes a matrix compound;

Preferably, the ointment base includes vegetable oil, animal fat, and semi-solid hydrocarbons obtained from petroleum, such as: lanolin, vaseline, liquid paraffin, mineral oil, etc.;

Preferably, the gel matrix compound is a polymer substance that has good biocompatibility and can form a gel, such as: poloxamer, gellan gum, chitosan, sodium alginate, sodium hyaluronate, Hypromellose, xanthan gum, carbomer, etc.;

Preferably, the formulations of the above pharmaceutical composition include common formulations, controlled release formulations, targeted formulations and the like.

The above-mentioned pharmaceutical composition can also be used in conjunction with other known corneal epithelial injury treatment methods and/or therapeutic agents. Another aspect of the present invention provides a synergistic pharmaceutical composition for treating and/or preventing corneal epithelial injury, including the above-mentioned drugs A composition and a therapeutic agent for corneal epithelial damage.

Preferably, the corneal epithelial injury includes persistent corneal epithelial injury, diabetic keratopathy, neurokeratitis, and limbal stem cell deficiency.

Another aspect of the present invention also provides a contact lens, which contains, carries, or is connected with the above-mentioned pharmaceutical composition, and patients can achieve the purpose of administration and treatment by wearing the contact lens.

The pharmaceutical composition provided by the invention can not only promote the proliferation and clone formation ability of limbal stem cells, but also effectively promote the repair of corneal epithelial damage, including the repair of persistent corneal epithelial damage, the treatment of diabetic keratopathy, and the treatment of neurokeratitis , The treatment of limbal stem cell deficiency. In addition, in the pharmaceutical composition provided by the present invention, GDNF and OPG are preferably of human origin, have little adverse reactions, and can greatly improve patient compliance.

detailed description

The effective components are present in an effective amount in the pharmaceutical composition of the present invention. As used herein, the term "effective amount" refers to an amount sufficient to treat (therapeutically or prophylactically) the target disorder when administered in an appropriate dosage regimen. For example, an effective amount is sufficient to reduce or ameliorate the severity, duration, or progression of the disorder being treated, prevent progression of the disorder being treated, cause regression of the disorder being treated, or enhance or improve another therapy preventive or therapeutic effect.

The term "treatment" includes both therapeutic treatment and prophylactic treatment (reducing the likelihood of development). The terms mean reducing, inhibiting, attenuating, shrinking, halting or stabilizing the development or progression of, lessening the severity of, or ameliorating symptoms associated with, a disease such as a disease or condition described herein.

Another embodiment of the present invention relates to pharmaceutical compositions contained within a swab or sponge that can be applied to the ocular surface.

The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

GDNF and OPG used in the present invention can be purchased commercially, and can also be extracted according to methods disclosed in the prior art, such as extracting from tissues such as male mice and human placenta, or prepared according to methods disclosed in the prior art, such as through gene Preparation by engineering methods, or use of GDNF or OPG drug substitutes disclosed in the prior art does not limit the scope of the present invention, although the GDNF and OPG used in the following examples are human recombinant GDNF and OPG.

Example 1 eye drops

Prepare eye drops according to the preparation method known in the art, and its main component and content are as follows:

Example 2 eye drops

Prepare eye drops according to the preparation method known in the art, and its main component and content are as follows:

Example 3 eye drops

Prepare eye drops according to the preparation method known in the art, and its main component and content are as follows:

Example 4 eye drops

Prepare eye drops according to the preparation method known in the art, and its main component and content are as follows:

Example 5 Ophthalmic Gel

Prepare ophthalmic gel according to the preparation method known in the art, its main component and content are as follows:

Example 6 Eye ointment

Prepare ophthalmic ointment according to the preparation method known in the art, its main component and content are as follows:

Example 7 Eye film

According to the preparation methods known in the art to prepare ophthalmic film, its main components and content are as follows:

Embodiment 8 drug efficacy test

1. Normal corneal epithelial injury repair test

1.1 Experimental materials and conditions

Test sample: the eye drops prepared by embodiment 1-4;

Comparative sample: removed the eye drops prepared by the embodiment 1-4 of OPG;

Control sample: normal saline;

Test animals: C57BL/6 mice, 6-8 weeks old.

1.2 Experimental method

The epithelium in the central 3 mm area of the mouse right cornea was scraped using a 3 mm trephine and an epithelial spatula, and the drug was administered. Using sodium fluorescein staining and taking pictures under a slit lamp, the epithelial injury repair of mice was observed.

1.3 Experimental results

The results showed that compared with the control sample, both the test sample and the comparison sample could effectively promote the repair of corneal epithelial damage, but the repair effect of the test sample group was significantly better than that of the comparison sample group.

2. STZ-induced diabetic corneal epithelial injury repair test

2.1 Experimental materials and conditions

Test sample: the eye drops prepared by embodiment 1-4;

Comparative sample: removed the eye drops prepared by the embodiment 1-4 of OPG;

Control sample: normal saline;

Test animals: STZ-induced C57BL/6 diabetic mice, 6-8 weeks old.

2.2 Experimental method

The epithelium in the central 3 mm area of the mouse right cornea was scraped using a 3 mm trephine and an epithelial spatula, and the drug was administered. Using sodium fluorescein staining and taking pictures under a slit lamp, the epithelial injury repair of mice was observed.

2.3 Experimental results

The results showed that compared with the control sample, both the test sample and the control sample could effectively promote the repair of STZ-induced diabetic corneal epithelial damage, but the repair effect of the test sample group was significantly better than that of the control sample group.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention within.

Claims (10)

1. A pharmaceutical composition for treating and/or preventing corneal epithelial damage, comprising GDNF in the pharmaceutical composition, and preferably, the pharmaceutical composition also contains OPG. 2. The pharmaceutical composition according to claim 1, wherein the GDNF is murine GDNF or human GDNF; and/or, the OPG is murine OPG or human OPG. 3. The pharmaceutical composition according to claim 1, wherein said human GDNF is human recombinant GDNF; and/or, said human OPG is human recombinant OPG. 4. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is a preparation for local administration, gastrointestinal administration or parenteral administration. 5. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition is a preparation for ophthalmic administration, preferably selected from the group consisting of eye drops, eye ointment, gel and ophthalmic film. 6. The pharmaceutical composition according to claim 5, wherein the pharmaceutical composition is an ophthalmic solution, including aqueous eye drops and non-aqueous eye drops. 7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition is aqueous eye drops, wherein the content of GDNF is 1ng/ml-1000μg/ml, preferably 10-5000ng/ml , more preferably 100-1000ng/ml; and/or, the content of OPG is 1ng/ml-100ng/ml, preferably 10-50ng/ml. 8. The pharmaceutical composition according to any one of claims 1-7, wherein the pharmaceutical composition also includes pharmaceutically acceptable additives; Preferably, the additive is selected from one or more of bacteriostats, pH regulators, osmotic pressure regulators, solubilizers, thickeners and chelating agents; Preferably, the bacteriostatic agent is selected from one or more of: thimerosal, quaternary ammonium salts, dumiphene, chlorhexidine, chlorobutanol, parabens and sorbic acid; Preferably, the preservative content in the pharmaceutical composition is 0.001-1.0wt%; Preferably, the pH regulator is selected from one or more of: phosphate, acetate, citric acid and its salts, carbonate solution, sodium hydroxide, potassium hydroxide, hydrochloric acid, boric acid and phosphoric acid ; Preferably, the osmotic pressure regulator is selected from one or more of sugars, polyalcohols and salts; Preferably, the solubilizer is selected from: one or more of cyclodextrin and its derivatives, water-soluble polymers and surfactants; Preferably, the thickener is selected from one of methylcellulose, carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose and salts thereof or more; Preferably, the chelating agent is selected from one or more of sodium edetate, sodium citrate and sodium condensed phosphate. 9. Use of GDNF alone or in combination with OPG in the preparation of a medicament for treating corneal epithelial damage (preferably diabetic corneal epithelial damage). 10. The use according to claim 9, wherein the medicament is eye drops.