Composition for reducing intraocular pressure and application

Abstract

The invention provides a composition for reducing intraocular pressure and application, and relates to the technical field of medical chemistry. The composition comprises potassium hyaluronate. It is proved for the first time that potassium hyaluronate can achieve the effect of reducing intraocular pressure by down-regulating expression of TIPARP.

Description

Technical Field

[0001] The present application relates to the technical field of pharmaceutical chemistry, and in particular to a composition for reducing intraocular pressure and its application. Background Art

[0002] Potassium hyaluronate is a potassium salt of hyaluronic acid evolved from sodium hyaluronate. It has good moisturizing, film-forming, lubricating and nourishing effects, anti-wrinkle and anti-aging effects, UV protection and repair capabilities. Hyaluronic acid and its salt products have been widely used in skin care, hair care, wound repair and other fields. However, the auxiliary antihypertensive effect of potassium hyaluronate has only been demonstrated in blood pressure lowering products, while its role in lowering intraocular pressure has not been widely developed.

[0003] In terms of lowering blood pressure, the effect of potassium hyaluronate mainly comes from potassium ions. Potassium ions can cause more sodium ions to be excreted from the urine through the blood via the sodium-potassium pump, thereby reducing the tightening of blood vessel walls. When blood vessel walls are relaxed, blood can flow more smoothly and lower blood pressure.

[0004] High intraocular pressure is an eye disease caused by the pressure of the aqueous humor on the cornea from the inside to the outside. The space inside the cornea of ​​the eye and outside the pupil is called the anterior chamber, which is filled with aqueous humor. Aqueous humor is continuously produced by the ciliary body and flows into the anterior chamber. After providing nutrition to the cornea and other parts of the eye, it flows out from the trabecular meshwork (uveal and corneoscleral meshwork) or Schlemm's canal. Usually, high intraocular pressure is caused by the inflow of aqueous humor being greater than the outflow of aqueous humor. This situation mainly occurs because the outflow of aqueous humor is blocked. The blood pressure reduction effect can be achieved by increasing the outflow of aqueous humor or reducing the secretion of aqueous humor.

[0005] Drugs that can reduce intraocular pressure mainly include prostaglandins, beta-blockers, etc. These drugs reduce intraocular pressure by increasing aqueous humor outflow or reducing aqueous humor production. However, these drugs are usually irritating, and long-term use will lead to dependence, which will reduce the body's own regulation of intraocular pressure. Summary of the invention

[0006] The purpose of the present invention is to provide a composition and application for reducing intraocular pressure. The present invention proves for the first time that potassium hyaluronate can reduce intraocular pressure by down-regulating the expression of TIPARP.

[0007] In one aspect, the present application provides a composition for reducing intraocular pressure, the composition comprising potassium hyaluronate (CAS#: 31799-91-4).

[0008] Furthermore, the molecular weight of the potassium hyaluronate is 1 KDa-2000 KDa.

[0009] Preferably, the molecular weight of the hyaluronic acid can be selected from any value of 1KDa, 2KDa, 3KDa, 4KDa, 5KDa, 6KDa, 7KDa, 8KDa, 9KDa, 10KDa, 20KDa, 30KDa, 40KDa, 50KDa, 60KDa, 70KDa, 80KDa, 90KDa, 100KDa, 200KDa, 300KDa, 400KDa, 500KDa, 600KDa, 700KDa, 800KDa, 900KDa, 1000KDa, 1500KDa, and 2000KDa.

[0010] Preferably, the molecular weight of the potassium hyaluronate is 1 KDa-500 KDa.

[0011] More preferably, the molecular weight of the potassium hyaluronate is 1 KDa-300 KDa.

[0012] Furthermore, the concentration of the potassium hyaluronate is 0.001%-5% by mass of the composition.

[0013] Furthermore, the concentration of the potassium hyaluronate is 0.005%-1% by mass of the composition.

[0014] Preferably, the concentration of potassium hyaluronate can be selected from any value of 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, and 5%.

[0015] The composition of the present application may also be added with excipients, which may be appropriate solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesive agents, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, etc.

[0016] The composition of the present application can be prepared by a general method, wherein one or more diluents or carriers can be added, such as pills, tablets, capsules, granules, powders, lozenges, syrups, emulsions, suspensions, etc.; preferably, eye drops.

[0017] Those skilled in the art will appreciate that the composition can be prepared using a general method.

[0018] In a preferred embodiment, the preparation method of the composition comprises: dissolving potassium hyaluronate in water and stirring evenly.

[0019] In a preferred embodiment, the composition comprises:

[0020] 0.001%-5% potassium hyaluronate with a molecular weight of 1KDa-2000 KDa, 0.01%-0.7% chitosan derivative, 0.01%-0.7% hydroxypropyl methylcellulose, 0.01%-1.5% osmotic pressure regulator, 0.01%-0.5% chelating agent, 0.005%-0.1% antibacterial agent, 0.001%-0.1% sodium perborate tetrahydrate, 0.5%-2% propylene glycol, 0.5%-5.5% boric acid, and the rest is water.

[0021] On the other hand, the present application also provides the use of potassium hyaluronate or the composition in inhibiting the expression of TCDD-induced poly (ADP-ribose) polymer (TIPARP).

[0022] On the other hand, the present application also provides the use of potassium hyaluronate or the composition in the preparation of a drug for reducing intraocular pressure.

[0023] Preferably, the lowering of intraocular pressure is achieved by down-regulating the expression of TIPARP.

[0024] The present invention has the following beneficial effects:

[0025] This application proves for the first time that potassium hyaluronate can reduce intraocular pressure by down-regulating the expression of TIPARP, providing a new effective ingredient for reducing intraocular pressure and a new preventive drug ingredient for diseases caused by high intraocular pressure. DETAILED DESCRIPTION

[0026] In order to more clearly illustrate the overall concept of the application, the following is described in detail by way of embodiment. In the following description, a large amount of specific details are provided to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0027] Among them, Lentivirus-GFP-TIPARP was purchased from Jiangsu Hanheng Medical Instrument Co., Ltd., and the trabecular meshwork cell model (HTM) was purchased from ScienCell (USA).

[0028] Unless otherwise specified, in the following embodiments, the reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.

[0029] If no specific conditions are specified in the examples, the experiments were carried out according to conventional conditions or conditions recommended by the manufacturer.

[0030] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in the field of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields, and can be specifically carried out according to Molecular Cloning: A Laboratory Manual (Fourth Edition).

[0031] In addition, the "water" mentioned in the present invention includes any feasible water that can be used in the pharmaceutical field, such as deionized water, distilled water, ion exchange water, double distilled water, high-purity water, and purified water.

[0032] The details of the reagents and instruments involved in the following examples are shown in Table 1.

[0033] Table 1 Reagents and instruments

[0034] name factory Potassium hyaluronate (purity> 99%) Bloomage Biopharmaceuticals Sodium hyaluronate (purity>99%) Bloomage Biopharmaceuticals <![CDATA[CO2 Incubator]]> Thermo, 160i ELISA reader Tecan, Spark Trabecular meshwork cells ScienCell High glucose DMEM medium VivaCell TIPARP ELISA Kit Human

[0035] Example 1

[0036] Weigh 0.0016 g of potassium hyaluronate (molecular weight 1800 KDa) powder and dissolve it in 99.9984 g of deionized water, and stir evenly until it is completely dissolved.

[0037] Example 2

[0038] Weigh 0.0016 g of potassium hyaluronate (molecular weight 1 KDa) powder and dissolve it in 99.9984 g of deionized water, stirring evenly until completely dissolved.

[0039] Example 3

[0040] Weigh 0.0016 g of potassium hyaluronate (molecular weight 300 KDa) powder and dissolve it in 99.9984 g of deionized water, stirring evenly until completely dissolved.

[0041] Comparative Example 1

[0042] Weigh 0.0016 g of sodium hyaluronate (molecular weight 1800 KDa) powder and dissolve it in 99.9984 g of deionized water, stirring evenly until completely dissolved.

[0043] Comparative Example 2

[0044] Weigh 0.0016 g of sodium hyaluronate (molecular weight 1 KDa) powder and dissolve it in 99.9984 g of deionized water, stirring evenly until completely dissolved.

[0045] Comparative Example 3

[0046] Weigh 0.0016 g of sodium hyaluronate (molecular weight 300 KDa) powder and dissolve it in 99.9984 g of deionized water, stirring evenly until it is completely dissolved.

[0047] Comparative Example 4

[0048] Weigh 0.0016 g of sodium hyaluronate (molecular weight 300 KDa) and 0.0094 g of potassium chloride powder and dissolve them in 99.9890 g of deionized water, stirring evenly until completely dissolved.

[0049] The specific preparation parameters of the above-mentioned examples and comparative examples are shown in Table 2.

[0050] Table 2 Different hyaluronic acid molecular weight and concentration

[0051]

[0052] Experimental example: blood pressure reduction effect test

[0053] There are documents that show that the expression of TIPARP is directly related to the regulation of intraocular pressure. Therefore, the effect of lowering intraocular pressure can be characterized by measuring the expression of TIPARP. The reduction in the expression of TIPARP can prove that the sample has the effect of lowering intraocular pressure (reference: Zhang Y, Song M, Bi Y, et al. TIPARP is involved in the regulation of intraocular pressure[J]. Communications Biology[2025-02-06]. DOI:10.1038 / s42003-022-04346-0.).

[0054] Trabecular meshwork cells were collected at a rate of 5 × 10 4 Inoculate cells into a 24-well plate at a seeding density of cells / well and incubate overnight in an incubator (37°C, 5% CO2). When the cell plating rate in the 24-well plate reaches 40%-60%, administer the drug. Add 1 mL of cell culture medium to each well of the blank control group; add 1 mL of culture medium containing the corresponding concentration of the test substance (Examples 1-3 and Comparative Examples 1-4) to each well of the sample group. After the incubation, collect the cells and lyse them for subsequent detection (Note: the amount of collected samples is determined according to the detection indicators). After collection, the samples are placed in a -80°C refrigerator for freezing. Detect the expression level of TIPARP according to the operating instructions of the Human TIPARP ELISA kit.

[0055] The test results are shown in Table 3.

[0056] Table 3 TIPARP expression

[0057] No TIPARP expression Blank control 8.124 Example 1 <![CDATA[3.712 ***# ]]> Example 2 <![CDATA[3.519 ***# ]]> Example 3 <![CDATA[3.647 **#& ]]> Comparative Example 1 <![CDATA[4.361 ** ]]> Comparative Example 2 <![CDATA[4.213 * ]]> Comparative Example 3 <![CDATA[4.398 * ]]> Comparative Example 4 <![CDATA[4.533 * ]]>

[0058] Note: *** indicates that compared with the blank control, p<0.001;

[0059] ** indicates, compared with blank control, p < 0.01;

[0060] * indicates, compared with blank control, p < 0.05;

[0061] # It indicates that there is a significant difference compared with sodium hyaluronate of the same molecular weight, p<0.05;

[0062] & It indicates that there is a significant difference compared with Comparative Example 4, p<0.05.

[0063] The results in Table 3 prove that potassium hyaluronate can significantly reduce the expression of TIPAPR (p < 0.01) relative to the blank control, and also significantly reduce the expression of TIPAPR relative to sodium hyaluronate of the same molecular weight and concentration, indicating that potassium hyaluronate has a significant effect of reducing intraocular pressure, and the effect is better than sodium hyaluronate. At the same time, potassium hyaluronate has a significant effect of reducing TIPARP relative to the same molar amount of sodium hyaluronate and potassium chloride combination, indicating that potassium hyaluronate has a significantly better intraocular pressure-reducing effect than a physical mixture of sodium hyaluronate and potassium chloride.

[0064] Example 4

[0065] Disclosed is an eye drop containing potassium hyaluronate, comprising 5% potassium hyaluronate, 0.01%-0.7% chitosan derivative, 0.01%-0.7% hydroxypropyl methylcellulose, 0.01%-1.5% osmotic pressure regulator, 0.01%-0.5% chelating agent, 0.005%-0.1% antibacterial agent, 0.001%-0.1% sodium perborate tetrahydrate, 0.5%-2% propylene glycol, 0.5%-5.5% boric acid, and the remainder being deionized water.

[0066] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. A composition for reducing intraocular pressure, characterized in that The composition includes potassium hyaluronate.

2. The composition according to claim 1, characterized in that The molecular weight of the potassium hyaluronate is 1KDa-2000KDa.

3. The composition according to claim 2, characterized in that The molecular weight of the potassium hyaluronate is 1KDa-500KDa.

4. The composition according to claim 3, characterized in that The molecular weight of the potassium hyaluronate is 1KDa-300KDa.

5. The composition according to claim 1, characterized in that Calculated by mass percentage of the composition, the concentration of potassium hyaluronate is 0.001%-5%.

6. The composition according to claim 5, characterized in that Calculated by mass percentage of the composition, the concentration of potassium hyaluronate is 0.005%-1%.

7. Use of potassium hyaluronate or the composition according to any one of claims 1 to 6 in inhibiting the expression of TCDD-inducible poly (ADP-ribose) polymerase.

8. Use of potassium hyaluronate or the composition according to any one of claims 1 to 6 in the preparation of a medicament for reducing intraocular pressure.

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