An ophthalmic pharmaceutical composition and its preparation method and application

Abstract

The present invention relates to the technical field of ophthalmic drugs, and more particularly to a kind of ophthalmic pharmaceutical composition and its preparation method and application. The active ingredient of the ophthalmic pharmaceutical composition provided by the present invention includes glutamine-metal ion complex, and glutamine-metal ion complex has significantly higher stability compared with glutamine, thereby being conducive to improving the stability of the pharmaceutical composition. Moreover, compared with the separate effects of glutamine and metal ions, glutamine and metal ions in glutamine-metal ion complex can act synergistically, significantly prolonging their retention time on the ocular surface, and having significantly better efficacy for dry eye, having both the advantages of improving stability, increasing ocular surface retention time and enhancing the therapeutic effect of dry eye, and having good application prospects in the treatment of eye diseases such as dry eye and drug preparation.

Description

Technical Field

[0001] The present invention relates to the technical field of ophthalmic medicines, and in particular to an ophthalmic medicine composition, a preparation method and an application thereof. Background Art

[0002] After dry eye injury, glutaminase 1 (GLS1) expression is upregulated, and corneal glutamine content is reduced. Glutamine can downregulate pro-inflammatory and pro-fibrotic corneal epithelial cell subsets in dry eye models, reduce inflammatory cytokine secretion, and promote corneal epithelial repair. Patent applications such as WO2010107069A1, CN114869873A, and US2021 / 0030833A1 disclose the use of amino acids, including glutamine, in the preparation of drugs for the treatment of dry eye.

[0003] However, the presence of an amide bond at the γ-position of glutamine makes it very unstable, and it is easily decomposed into pyroglutamic acid or glutamic acid under acidic, alkaline or heated conditions. Moreover, glutamine has a single mechanism of action in the treatment of dry eyes and its effect is limited. Conventional glutamine eye drops have problems such as short residence time in the eye, easy dilution by tears, and low bioavailability. Summary of the Invention

[0004] The present invention provides an ophthalmic pharmaceutical composition, a preparation method and application thereof.

[0005] Specifically, the present invention provides the following technical solutions.

[0006] In a first aspect, the present invention provides an ophthalmic pharmaceutical composition comprising an active ingredient and an osmotic pressure regulator, wherein the active ingredient comprises a glutamine-metal ion complex.

[0007] Although metal ions (such as zinc and selenium) inherently possess antibacterial, anti-inflammatory, and tear film stabilization properties, their direct addition can easily cause eye irritation or precipitation with drug ingredients. During the development process, the present invention discovered that glutamine-metal ion complexes, formed by coordinating glutamine with metal ions, not only significantly improve glutamine's stability, but also create a synergistic effect between glutamine and metal ions in alleviating dry eye symptoms, promoting tear secretion, and repairing the cornea. Furthermore, glutamine-metal ion complexes can prolong the retention of active ingredients on the ocular surface, enhancing their efficacy. Experimental results show that glutamine-metal ion complexes are significantly more effective than glutamine and metal ions alone in alleviating dry eye symptoms, promoting tear secretion, and repairing the cornea, demonstrating significantly superior efficacy in the prevention and treatment of dry eye.

[0008] As an embodiment of the present invention, the glutamine-metal ion complex is a complex formed by glutamine or its derivatives and metal ions.

[0009] In the present invention, glutamine can be L-glutamine or D-glutamine. Glutamine derivatives are substances derived from glutamine that can release glutamine in vivo and coordinate with metal ions, such as alanylglutamine.

[0010] The carboxylic acid group (-COOH), amino group (-NH2), and side chain amide group (-CONH2) in the glutamine molecule can all serve as coordination sites for metal ions (generally with a coordination number of 2 or 3), thereby forming complexes with metal ions through coordination bonds. Although glutamine is known to form complexes with metal ions, its ocular activity and efficacy in treating ocular diseases such as dry eye are difficult to predict, and there are no reports on the use of glutamine-metal ion complexes in the preparation of ocular drugs. The present invention significantly improves the stability of ophthalmic medications such as glutamine-containing eye drops by forming a water-soluble complex with a metal ion through coordination bonds. Furthermore, in the glutamine-metal ion complex, the coordination of glutamine with the metal ion can produce a synergistic therapeutic effect through a dual mechanism of metal ion and glutamine. Glutamine reduces the secretion of inflammatory cytokines and promotes corneal epithelial repair. Metal ions can pass through cell membranes using ion channels formed by membrane-bound peptides and proteins, or they can be transported into cells via transporters. Coordination of glutamine with metal ions allows for faster intracellular transport with the metal ion, increasing drug retention time and enhancing efficacy. Furthermore, metal ions can inhibit tear evaporation (by regulating the lipid layer), reduce inflammatory factors (such as IL-6 and TNF-α), and inherently possess certain antibacterial and anti-inflammatory effects, as well as maintain tear film stability. After coordination, they can exert a synergistic therapeutic effect with glutamine, enhancing the therapeutic effect for dry eye. Preferably, the metal ion is a divalent metal ion or a metal ion with a valence higher than divalent.

[0011] The metal ions with a valence higher than divalent can be trivalent, tetravalent, pentavalent, or hexavalent metal ions.

[0012] Wherein, the divalent metal ion is preferably selected from Zn² + Mg² + 、Se² + 、Cu² + The coordination number of the above-mentioned divalent metal ion is usually 4 or 6.

[0013] Preferably, for the complex formed by glutamine or its derivatives and metal ions, the molar ratio of glutamine or its derivatives to the metal ions is 2:1 to 1:1.

[0014] In some embodiments of the present invention, the metal ion is Zn² + , that is, the glutamine-metal ion complex is glutamine-Zn² +Complex. Glutamine can react with Zn² + Coordinated to form a stable complex, compared with glutamine, glutamine-Zn² + The stability of the complex to high temperature and light is significantly improved, thereby improving the stability of the pharmaceutical composition. In addition, compared with glutamine, glutamine-Zn² + The complex significantly improves the therapeutic effect of dry eyes.

[0015] As another embodiment of the present invention, the glutamine-metal ion complex is a ternary asymmetric coordination structure formed by the metal ion, glutamine or its derivative and a second metal ligand.

[0016] Wherein, the second metal ligand is ethylenediaminetetraacetic acid or its salt, or citric acid.

[0017] The present invention unexpectedly discovered that a ternary asymmetric coordination structure formed by simultaneously coordinating a metal ion with glutamine (i.e., the first metal ion ligand) and a second metal ligand can further enhance stability to high temperatures and light. More importantly, it can slowly release glutamine and metal ions, prolonging the duration of drug action and improving drug efficacy. Its efficacy in treating dry eye is significantly enhanced compared to the complex formed by glutamine and metal ions described above. When administered to animals with dry eye, this ternary complex exhibits excellent effects in promoting tear secretion and repairing corneal damage.

[0018] Preferably, the metal ion is a divalent metal ion or a metal ion with a valence higher than divalent;

[0019] The metal ions with a valence higher than divalent can be trivalent, tetravalent, pentavalent, or hexavalent metal ions.

[0020] Wherein, the divalent metal ion is preferably selected from Zn² + Mg² + 、Se² + 、Cu² + At least one of. More preferably Zn² + .

[0021] Preferably, in the above ternary asymmetric coordination structure, the molar ratio of glutamine or its derivative to the second metal ligand is 4:1-1:1; the molar ratio of the total amount of glutamine or its derivative and the second metal ligand to the metal ion is 2:1-1:1.

[0022] In some embodiments of the present invention, the metal ion is Zn² + , that is, the glutamine-metal ion complex is glutamine-Zn² +-ethylenediaminetetraacetic acid or its salt ternary complex (e.g. glutamine-Zn² + -EDTA-2Na ternary complex), or glutamine-Zn² + -Citric acid ternary complex. When administered to animals with dry eye, the ternary complex demonstrated excellent effects in promoting tear secretion and repairing corneal damage, significantly outperforming glutamine alone or other glutamine ternary complexes.

[0023] In the above-mentioned eye drops, the concentration of the glutamine-metal ion complex is 0.3% to 5% based on the glutamine concentration.

[0024] According to the glutamine concentration meter, the concentration of glutamine-metal ion complex is converted into the concentration of glutamine.

[0025] In the present invention, the glutamine-metal ion complex can be prepared by conventional methods.

[0026] For example, when the glutamine-metal ion complex is a complex formed by glutamine and a metal ion, the preparation method of the glutamine-metal ion complex comprises: dissolving glutamine and a metal ion salt in water, and stirring the mixture at 55-65° C. The molar ratio of glutamine to the metal ion is preferably 10:1 to 1:1.

[0027] When the glutamine-metal ion complex is a ternary asymmetric coordination structure formed by the metal ion, glutamine, and a second metal ligand, the preparation method of the glutamine-metal ion complex comprises: mixing and dissolving glutamine, a metal ion salt, and a second metal ligand in water, and stirring the mixture at 55-65°C. The molar ratio of glutamine, the metal ion salt, and the second metal ligand is (1-2):(1-2):(0.4-1). After the mixture is dissolved in water, the concentration of the second metal ligand is 0.05%-3%.

[0028] The stirring reaction time may be 1-2 hours.

[0029] The active ingredients of the ophthalmic pharmaceutical composition of the present invention may include, in addition to the glutamine-metal ion complex, other active ingredients for preventing or treating dry eye, or active ingredients for preventing or treating other eye diseases, or anti-inflammatory and antibacterial active ingredients.

[0030] Examples of the other active ingredients mentioned above for preventing or treating dry eye include: polyvinyl alcohol, sodium hyaluronate, artificial tears in gel form, cyclosporine A (CsA), tacrolimus, etc., and active ingredients of traditional Chinese medicine such as xiasangju can also be used.

[0031] In the ophthalmic pharmaceutical composition of the present invention, the osmotic pressure regulator includes at least one selected from sodium chloride, mannitol, glycerol, glucose, boric acid, borax, potassium chloride, and sorbitol.

[0032] Osmotic pressure regulators (also known as isotonic regulators or isotonic agents) are common components of ophthalmic drugs. The present invention has no particular limitation on the specific selection of such components.

[0033] Since ophthalmic drugs such as eye drops should be isotonic with tears, the amount of osmotic pressure regulator added should be based on the amount that can achieve isotonicity between the pharmaceutical composition and tears. The specific amount can be adjusted according to the selected osmotic pressure regulator.

[0034] Preferably, the concentration of the osmotic pressure regulator in the ophthalmic pharmaceutical composition is 0.5%-5%, preferably 4%-5%.

[0035] In some embodiments of the present invention, the osmotic pressure regulator is mannitol. The concentration of mannitol in the pharmaceutical composition is 4-5%.

[0036] In order to adjust the pH of the ophthalmic pharmaceutical composition to make it closer to physiological pH, the ophthalmic pharmaceutical composition of the present invention further comprises a pH buffer.

[0037] Wherein, the pH buffer comprises at least one selected from phosphate buffer, borate buffer, sodium hydroxide, hydrochloric acid, boric acid buffer, and citrate buffer.

[0038] Preferably, the phosphate buffer is disodium hydrogen phosphate and sodium dihydrogen phosphate, or dipotassium hydrogen phosphate and potassium dihydrogen phosphate.

[0039] The amount of the pH buffer added is based on the ability to maintain the pH of the pharmaceutical composition within the physiological pH range (pH 5-8, preferably pH 6.3-7.3, more preferably 6.4-6.8).

[0040] In some embodiments of the present invention, the pH buffer is disodium hydrogen phosphate and sodium dihydrogen phosphate. In the pharmaceutical composition, the concentration of disodium hydrogen phosphate is 0.4-0.6%, and the concentration of sodium dihydrogen phosphate is 0.1-0.2%.

[0041] The ophthalmic pharmaceutical composition of the present invention may further comprise a solvent, preferably water.

[0042] The ophthalmic pharmaceutical composition of the present invention may optionally contain a polymeric material. This addition can further prolong the drug's retention time on the ocular surface. The polymeric material may include one or more of sodium hyaluronate (molecular weight 10-500 kDa), chitosan derivatives (such as carboxymethyl chitosan), hypromellose, polyethylene glycol, polycarbophil, and polyvinyl alcohol. The concentration of the polymeric material in the pharmaceutical composition is preferably 0.01%-10%, more preferably 0.1%-2%.

[0043] The ophthalmic pharmaceutical composition of the present invention may optionally contain a preservative. There are no particular limitations on the type of preservative; all preservatives permitted for ophthalmic use may be used, such as benzalkonium chloride. The concentration of the preservative is preferably 0.005% to 0.01%.

[0044] In a second aspect, the present invention provides use of the above-mentioned ophthalmic pharmaceutical composition in the preparation of a medicament for preventing and / or treating ophthalmic diseases.

[0045] Preferably, the ophthalmic disease comprises dry eye.

[0046] The drug can be a liquid preparation (such as eye drops, intraocular injection), a semi-solid preparation (eye gel, eye ointment, eye cream, etc.), or a solid preparation (eye pills, etc.).

[0047] In a third aspect, the present invention provides an ophthalmic drug comprising the above-mentioned ophthalmic pharmaceutical composition.

[0048] In a fourth aspect, the present invention provides a method for preparing the above-mentioned ophthalmic pharmaceutical composition, the method comprising: mixing the glutamine-metal ion complex with an osmotic pressure regulator.

[0049] Preferably, when the glutamine-metal ion complex is a complex formed by glutamine and a metal ion, the preparation method of the glutamine-metal ion complex comprises: dissolving glutamine and a metal ion salt in water, and stirring the mixture at 55-65°C for reaction.

[0050] When the glutamine-metal ion complex is a ternary asymmetric coordination structure formed by the metal ion, glutamine and a second metal ligand, the preparation method of the glutamine-metal ion complex comprises: dissolving glutamine, a metal ion salt and a second metal ligand in water, and stirring the mixture at 55-65° C. to react.

[0051] In a fifth aspect, the present invention provides use of a glutamine-metal ion complex in the preparation of an ophthalmic pharmaceutical composition;

[0052] Wherein, the glutamine-metal ion complex is a complex formed by glutamine or its derivatives and metal ions;

[0053] Alternatively, the glutamine-metal ion complex is a ternary asymmetric coordination structure formed by the metal ion, glutamine or its derivative and a second metal ligand; wherein the second metal ligand is ethylenediaminetetraacetic acid or its salt, or citric acid.

[0054] Preferably, the metal ion is a divalent metal ion or a metal ion with a valence higher than divalent;

[0055] Preferably, the divalent metal ion is selected from Zn² + Mg² + 、Se² + 、Cu² + At least one of; preferably Zn² + .

[0056] For the complex formed by glutamine or its derivatives and metal ions, the molar ratio of glutamine or its derivatives to the metal ions is preferably 2:1 to 1:1.

[0057] For the above-mentioned ternary asymmetric coordination structure, the molar ratio of glutamine or its derivative to the second metal ligand is 4:1~1:1; the molar ratio of the total amount of glutamine or its derivative and the second metal ligand to the metal ion is 2:1~1:1.

[0058] All the contents of the first aspect above apply to the above-mentioned glutamine-metal ion complex.

[0059] The beneficial effects of the present invention include at least one aspect: the active ingredient of the ophthalmic pharmaceutical composition provided by the present invention comprises a glutamine-metal ion complex. Compared to glutamine, the glutamine-metal ion complex exhibits significantly greater stability to high temperatures and light, thereby improving the stability of the pharmaceutical composition. Furthermore, compared to the individual effects of glutamine and the metal ion, the glutamine and metal ion in the glutamine-metal ion complex can act synergistically, significantly prolonging its retention time on the ocular surface, resulting in significantly superior therapeutic effects for dry eye. Combining the advantages of improved stability, increased ocular surface retention time, and enhanced dry eye treatment efficacy, the glutamine-metal ion complex has promising application prospects in the treatment of ocular diseases such as dry eye and in pharmaceutical preparation. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] In order to more clearly illustrate the technical solutions in the present invention or the prior art, a brief introduction is given below to the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0061] Figure 1 This is the infrared spectrum scanning result of the glutamine solution in Experimental Example 1 of the present invention.

[0062] Figure 2 This is the infrared spectrum scanning result of the complex solution of Example 1 in Experimental Example 1 of the present invention.

[0063] Figure 3 This is the infrared spectrum scanning result of the EDTA-2Na solution in Experimental Example 1 of the present invention.

[0064] Figure 4 This is the infrared spectrum scanning result of the complex solution of Example 3 in Experimental Example 1 of the present invention. DETAILED DESCRIPTION

[0065] The key terms and technical abbreviations used in this invention are explained as follows:

[0066] GLS1: glutaminase 1.

[0067] The Schirmer test quantifies tear production by placing a standardized filter paper strip in the conjunctival sac and measuring the length of the filter paper that tears soak over a certain period of time. It is an important tool in the diagnosis of dry eye.

[0068] EDTA-2Na: disodium ethylenediaminetetraacetic acid.

[0069] To make the objectives, technical solutions, and advantages of the present invention more clear, the following will be a clear and complete description of the technical solutions in the present invention. Obviously, the embodiments described are part of the embodiments of the present invention, not all of them. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0070] Example 1: Glutamine-Zn² + Preparation of complex eye drops (1)

[0071] This embodiment provides a glutamine-Zn² + The preparation method of the complex eye drops comprises the following steps: dissolving glutamine and zinc chloride in deionized water at a molar ratio of 2:1, stirring at 60°C for 2 hours, and obtaining glutamine-Zn². + The complex solution contains 0.6% glutamine.

[0072] Take the glutamine-Zn² prepared above +To 50 mL of the complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0073] Example 2: Glutamine-Zn² + Preparation of complex eye drops (2)

[0074] This embodiment provides a glutamine-Zn² + The preparation method of the complex eye drops comprises the following steps: dissolving glutamine and zinc chloride in deionized water at a molar ratio of 1:1, stirring at 60°C for 1 hour, and obtaining glutamine-Zn². + The complex solution contains 10.0% glutamine.

[0075] Take the glutamine-Zn² prepared above + To 50 mL of the complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0076] Example 3: Glutamine-Zn² + -EDTA-2Na complex eye drops preparation (1)

[0077] This embodiment provides a glutamine-Zn² + -EDTA-2Na complex eye drops, the preparation method thereof comprises the following steps: dissolving glutamine, zinc chloride and EDTA-2Na in deionized water at a molar ratio of 1:1:1, stirring at 60°C for 2 hours, and preparing glutamine-Zn² + -EDTA-2Na ternary complex solution, wherein the glutamine concentration is 2.0%.

[0078] Take the glutamine-Zn² prepared above + To 50 mL of EDTA-2Na ternary complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0079] Example 4: Glutamine-Zn² + -EDTA-2Na complex eye drops preparation (2)

[0080] This embodiment provides a glutamine-Zn² +-EDTA-2Na complex eye drops, the preparation method thereof comprises the following steps: dissolving glutamine, zinc chloride and EDTA-2Na in deionized water at a molar ratio of 1:2:1, stirring at 60°C for 2 hours, and preparing glutamine-Zn² + -EDTA-2Na ternary complex solution, wherein the glutamine concentration is 0.6%.

[0081] Take the glutamine-Zn² prepared above + To 50 mL of EDTA-2Na ternary complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0082] Example 5: Glutamine-Zn² + - Preparation of citric acid complex eye drops (1)

[0083] This embodiment provides a glutamine-Zn² + -citric acid complex eye drops, the preparation method thereof comprises the following steps: dissolving glutamine, zinc sulfate and citric acid in deionized water at a molar ratio of 1:1:1, stirring at 60°C for 1.5 hours, and preparing glutamine-Zn² + -citric acid ternary complex solution, wherein the glutamine concentration is 5.0%.

[0084] Take the glutamine-Zn² prepared above + -To 50 mL of citric acid ternary complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0085] Example 6: Glutamine-Zn² + - Preparation of citric acid complex eye drops (2)

[0086] This embodiment provides a glutamine-Zn² + -citric acid complex eye drops, the preparation method thereof comprises the following steps: dissolving glutamine, zinc sulfate and citric acid in deionized water at a molar ratio of 1.6:1:0.4, stirring at 60°C for 2 hours, and preparing glutamine-Zn² + -Citrate ternary complex solution, in which the glutamine concentration is 0.6%.

[0087] Take the glutamine-Zn² prepared above +-To 50 mL of citric acid ternary complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0088] Comparative Example 1: Glutamine-Zn² + Preparation of ethylenediamine complex eye drops

[0089] This comparative example provides a glutamine-Zn² + -ethylenediamine complex eye drops, the preparation method thereof comprises the following steps: dissolving glutamine, zinc chloride and ethylenediamine in deionized water at a molar ratio of 1:2:1, stirring at 60°C for 2 hours, and preparing glutamine-Zn² + -ethylenediamine ternary complex solution, wherein the glutamine concentration is 0.6%.

[0090] Take the glutamine-Zn² prepared above + To 50 mL of ethylenediamine ternary complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0091] Comparative Example 2: Glutamine-Zn² + -Preparation of sodium tripolyphosphate complex eye drops

[0092] This comparative example provides a glutamine-Zn² + -Sodium tripolyphosphate complex eye drops, the preparation method of which comprises the following steps: dissolving glutamine, zinc chloride and sodium tripolyphosphate in deionized water at a molar ratio of 1:2:1, stirring at 60°C for 2 hours, and preparing glutamine-Zn² + -Sodium tripolyphosphate ternary complex solution, in which the glutamine concentration is 0.6%.

[0093] Take the glutamine-Zn² prepared above + -To 50 mL of sodium tripolyphosphate ternary complex solution, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtration and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0094] Comparative Example 3: Preparation of glutamine eye drops

[0095] Take 0.3 g of glutamine, add 80 mL of water for injection, stir until dissolved, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtering and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0096] Comparative Example 4: Preparation of zinc chloride eye drops

[0097] Take 0.14 g of zinc chloride, add 80 mL of water for injection, stir until dissolved, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtering and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0098] Comparative Example 5: Glutamine eye drops containing zinc chloride

[0099] Take 0.3 g of glutamine and 0.14 g of zinc chloride, add 80 mL of water for injection, stir until dissolved, add 4.5 g of mannitol, 0.51 g of disodium hydrogen phosphate, and 0.17 g of sodium dihydrogen phosphate, and finally add water for injection to 100 mL. After filtering and sterilization, dispense into low-density polyethylene single-dose eye drop bottles.

[0100] Experimental Example 1: Infrared spectroscopy characterization of the complex

[0101] Take glutamine solution, EDTA-2Na solution, and the complex solution prepared in Example 1 and Example 3 respectively, and stir at 400~1000cm -1 Infrared spectrum scanning is performed within the wavenumber range, and the results are as follows Figure 1 、 Figure 2 、 Figure 3 and Figure 4 The characteristic peaks in the infrared spectral fingerprint region are more complex and not as obvious as those in the functional group region, but they are more sensitive to subtle changes in molecular structure. Subtle structural changes can lead to significant differences in the fingerprint region. Figure 1 and Figure 2 , Figure 2 Medium 850~900cm -1 , 750~800cm -1 550~600cm -1 The peak shape and intensity changed significantly within the wavenumber range, indicating that glutamine-Zn 2+ Complex formation. Compare Figure 1 、 Figure 3 and Figure 4 , Figure 4 Medium 850~950cm -1 The absorption peaks on the left and right changed significantly in shape and intensity, indicating that the structures of glutamine and EDTA-2Na changed, confirming the formation of a ternary complex.

[0102] Experimental Example 2: Stability Test

[0103] For the eye drops in the above embodiments and comparative examples, the stability of the eye drops was tested by a high temperature influencing factor test. The eye drops in the above embodiments 1 to 6 and comparative examples 1 to 3 were placed at a high temperature of 60°C for 30 days, and samples were taken on the 10th and 30th days to detect changes in the properties, pH value, content and related substances of the eye drops. The content of glutamine in each eye drop was determined using high-performance liquid chromatography (HPLC). The content (%) was calculated as follows: detected glutamine concentration / labeled glutamine concentration (theoretical concentration) × 100%. The content change (%) was calculated as follows: content at 10 or 30 days minus content at day 0. Related substance testing involved glutamine-related impurities using HPLC. The maximum single impurity (%) and total impurities (%) were calculated as the percentage of the peak area of ​​the largest detected single impurity and the sum of the impurity peak areas to the glutamine peak area, respectively. The maximum single impurity change (%) was calculated as follows: maximum single impurity at 10 or 30 days minus maximum single impurity at day 0. The total impurity change (%) was calculated as follows: total impurities at 10 or 30 days minus total impurities at day 0. The results are shown in Table 1.

[0104] Table 1 Stability test results

[0105]

[0106] The above results show that the stability of glutamine complexes with metal ions increases significantly. Among them, the ternary complex formed by glutamine, zinc ion, EDTA-2Na and citric acid has the best stability, while the ternary complex formed with ethylenediamine and sodium tripolyphosphate has poor stability.

[0107] Experimental Example 3: Treatment Effects on Dry Eye Animal Model

[0108] Experiments were conducted using a dry eye model induced by desiccation stress in mice to test the therapeutic efficacy of the eye drops described in the above examples and comparative examples. The successfully modeled experimental animals were divided into 11 groups of 6 animals each, consisting of a model control group, a blank excipient group, and 9 experimental groups. Six unmodeled animals were also selected as a normal control group. Grouping and dosing were performed as shown in Table 2. Tear secretion (Schirmer test) and corneal sodium fluorescein staining scores were measured before modeling (D-1), on the 7th day after dosing (D7), on the 14th day after dosing (D14), and on the 21st day after dosing (D21). The results are shown in Tables 3 and 4.

[0109] Table 2 Grouping and dosing schedule

[0110]

[0111] The tear secretion results are shown in Table 3. The corneal sodium fluorescein staining score results are shown in Table 4.

[0112] Table 3 Summary of tear secretion (mm)

[0113]

[0114] Note: Compared with the normal control group, a express p ≤0.05; compared with the blank excipient group, b express p ≤0.05; compared with experimental group 1, c express p ≤0.05.

[0115] Table 4 Summary of corneal fluorescein sodium staining scores

[0116]

[0117] Note: Compared with the normal control group, a express p ≤0.05; compared with the blank excipient group, b express p ≤0.05; compared with experimental group 1, c express p ≤0.05.

[0118] The above tear secretion volume and corneal sodium fluorescein staining results both show that glutamine-metal ion complex has a better therapeutic effect on dry eye, which is significantly better than glutamine eye drops and zinc chloride eye drops.

[0119] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. An ophthalmic pharmaceutical composition, characterized in that: The invention comprises an active ingredient and an osmotic pressure regulator, wherein the active ingredient comprises a glutamine-metal ion complex; Wherein, the glutamine-metal ion complex is a complex formed by glutamine or its derivatives and metal ions; Alternatively, the glutamine-metal ion complex is a ternary asymmetric coordination structure formed by the metal ion, glutamine or its derivative and a second metal ligand, wherein the second metal ligand is ethylenediaminetetraacetic acid or its salt, or citric acid; The metal ion is selected from Zn² + Mg² + 、Cu² + At least one of .

2. The ophthalmic pharmaceutical composition according to claim 1, characterized in that In the complex formed by glutamine or its derivative and the metal ion, the molar ratio of glutamine or its derivative to the metal ion is 2:1 to 1:

1.

3. The ophthalmic pharmaceutical composition according to claim 1, characterized in that In the ternary asymmetric coordination structure, the molar ratio of glutamine or its derivative to the second metal ligand is 4:1-1:1; the molar ratio of the total amount of glutamine or its derivative and the second metal ligand to the metal ion is 2:1-1:

1.

4. The ophthalmic pharmaceutical composition according to any one of claims 1 to 3, characterized in that In the ophthalmic pharmaceutical composition, the concentration of the glutamine-metal ion complex is 0.3% to 5% based on the concentration of glutamine or its derivatives.

5. The ophthalmic pharmaceutical composition according to any one of claims 1 to 3, characterized in that The osmotic pressure regulator includes at least one selected from sodium chloride, mannitol, glycerol, glucose, boric acid, borax, potassium chloride, and sorbitol.

6. The ophthalmic pharmaceutical composition according to any one of claims 1 to 3, characterized in that The ophthalmic pharmaceutical composition further comprises a pH buffer; Wherein, the pH buffer comprises at least one selected from phosphate buffer, borate buffer, sodium hydroxide, hydrochloric acid, boric acid buffer, and citrate buffer.

7. Use of the ophthalmic pharmaceutical composition according to any one of claims 1 to 6 in the preparation of medicaments for preventing and / or treating ophthalmic diseases.

8. The method for preparing the ophthalmic pharmaceutical composition according to any one of claims 1 to 6, characterized in that: The method comprises: mixing a glutamine-metal ion complex with an osmotic pressure regulator.

9. Application of glutamine-metal ion complexes in the preparation of ophthalmic pharmaceutical compositions; in, The glutamine-metal ion complex is a complex formed by glutamine or its derivatives and metal ions; Alternatively, the glutamine-metal ion complex is a ternary asymmetric coordination structure formed by the metal ion, glutamine or its derivative and a second metal ligand, wherein the second metal ligand is ethylenediaminetetraacetic acid or its salt, or citric acid; The metal ion is selected from Zn² + Mg² + 、Cu² + At least one of .

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