CN117815164A - Ophthalmic preparation for treating demodex mites - Google Patents

Abstract

The invention provides a nitroimidazole in-situ gel preparation for eyes, which is used for treating eye diseases caused by Demodex. Nitroimidazoles include, but are not limited to, metronidazole, tinidazole, ornidazole; the in-situ gel specifically comprises a temperature sensitive type, a pH sensitive type and an ion sensitive type, and is characterized in that after the in-situ gel is acted on affected parts of eyes by an eye preparation consisting of an active ingredient, an in-situ gel matrix and other auxiliary materials, the preparation is changed into a semisolid gel from a liquid state within a period of time under the influence of different environments or conditions, has certain viscosity, can stay on eyes for a certain period of time, and overcomes the defects of the traditional eye preparation and a post-improvement type preparation.

Description

Ophthalmic preparation for treating demodex mites

Technical Field

The invention belongs to the field of pharmaceutical preparations, relates to an ophthalmic preparation for treating demodex, and in particular relates to a nitroimidazole in-situ gel ophthalmic preparation for treating demodex blepharitis.

Background

Demodex was first found in cerumen in 1842, and Demodex was found in 1967 as one of the factors causing blepharitis, and specific cases of Demodex infecting eyelashes were first reported in 1984. Demodex is a common permanent small parasitic mite, and two kinds of Demodex are parasitic on eyes of human bodies, specifically, demodex folliculorum with the length of 0.35-0.4mm which is colonized on eyelash hair follicles, and Demodex sebaceous with the length of 0.2-0.3mm which is colonized on eyelash related sebaceous glands and meibomian glands. Parasitic life cycle of Demodex mites has 5 stages, respectively: eggs (60 h), larvae (36 h), pre-nymphs (72 h), nymphs (60 h) and adults (120 h), the whole life cycle is short, and the whole life cycle is about 15 days. Because of its short life cycle, the mating between male and female plays a non-negligible role in the infection process, demodex mites feed on skin keratinocytes and sebum secretions, mating is often carried out at night in the follicular orifice, the breeding level peaks after adequate food supply is obtained, and during this time facial Demodex mites also spread to the eyes, exacerbating the eye infection level.

Demodex can infect human body by directly contacting skin of a person infected with Demodex or indirectly contacting clothes, towels, dust containing Demodex, etc., and has close relation with personal health condition, sebum secretion condition, and cosmetic use, and when host immunity is low, demodex incidence rate will be increased. Because the transmission way is wide, and the parasitic disease does not occur in most of time, and is closely related to personal hygiene habit and autoimmunity of a host, it can be concluded that the human diseases caused by the demodex mites cannot be radically cured.

Among demodex mites which are lodged in eyes of a human body, demodex folliculorum lodges in eye eyelash hair follicles, and eats hair follicle epithelial cells and metabolites, which cause hair follicle expansion, hyperplasia, excessive keratinization, scaling at the root of eyelash, and cause eyelash shedding, trichiasis, mess eyelash, and the like, and the symptoms are called anterior blepharitis; another demodex that is lodged in the eyes of humans, demodex sebaceous colonizes the meibomian glands and their open areas, causing obstruction of the meibomian gland orifices, leading to meibomian gland dysfunction, and thus affecting tear film stability. The meibomian gland dysfunction caused by Demodex sebaceous mailing is considered by the scholars to be one of the causes of dry eye formation. In addition to dry eye, meibomian gland dysfunction can also cause meibomian gland cysts, and the mite body exterior formed by the Demodex sebaceous mites is covered with a hard exoskeleton that presents a foreign body sensation to the eye, which can thereby cause granulomatous responses, which in turn lead to aragonite. The above symptoms are called posterior blepharitis; blepharoconjunctivitis that can be caused when inflammation of the blepharoconjunctival margin caused by demodex spreads to the conjunctiva; and the trichiasis caused by Demodex mite infection causes punctate loss of corneal epithelium, which can form corneal ulcers for a long time and possibly cause keratitis. Besides the pathogenicity of Demodex itself, the bodies can also be used as a medium for other pathogens. Typical clinical manifestations are itching eyes, feeling of foreign matter on eyes, dry eyes, congestion of eyelid margin, scales, oversleeve-like secretion at the root of eyelashes, etc. The damage of the disease to the eyelid margin is large, and repeated attacks are easy to occur, so that lipid secretion is reduced, tear film stability is reduced, and various diseases are further caused. At present, no medicine for treating the disease is available in the market in China.

Conventional eye drops cannot kill Demodex mites, and ophthalmic preparations containing tea tree essential oil components are generally used for treating blepharitis caused by Demodex mites in the prior art. The existing literature experiments indicate that tea tree essential oil with the concentration of more than 15% can effectively kill demodex mites, but tea tree essential oil with the concentration of 10% burns eyes and damages corneas, so that the concentration of tea tree essential oil agents on the market does not exceed 5%, and the mite killing efficiency is greatly reduced.

Chinese patent 2022104012873 discloses a metronidazole ophthalmic gel preparation for treating eye infection caused by mites and acanthamoeba, which is characterized in that seaweed extract is used in preparation auxiliary materials, and the seaweed extract has the special functions of resisting high temperature, improving the administration comfort and preserving moisture. Although the preparation has the characteristics, the preparation still has the defects of the traditional ophthalmic preparation, namely, the medicine has short residence time in eyes, poor stability, is easily influenced by tears to further cause low bioavailability, and the common hydrogel has the problems of poor spreadability and difficult dosage control.

Chinese patent 021095035 discloses an ophthalmic preparation comprising poloxamer 188 and poloxamer 407 as temperature sensitive matrix, wherein the active ingredients are selected from anti-glaucoma drugs; antibiotic drugs; an antibacterial agent; one or a compound of antiviral drugs, but no embodiment supports a specific prescription of combining nitroimidazole drugs with a temperature-sensitive matrix, the disclosure of the patent is insufficient, the content of the specification cannot support the protection scope of the claims, and it cannot be proved that the nitroimidazole drugs can be combined with the temperature-sensitive matrix for use, or the preparation prescription with the feasibility is not designed, or the preparation has technical difficulty, and the problems to be solved in the research and development of the temperature-sensitive preparation are the above.

At present, novel preparations which are researched at home and abroad, such as ointment, suspension, hydrogel, implant and the like, have the defects of eye ointment taking Vaseline as a matrix, have strong greasy feeling and are easy to cause blurred vision after being used, although the retention time is prolonged to different degrees; the hydrogel has better biocompatibility, can prolong the acting time of the medicine, but the administration dosage is not easy to control; at present, the compliance of patients to implants is poor clinically, and the improvement mode still cannot effectively solve the defects of the traditional ophthalmic preparation.

Disclosure of Invention

After the defects of the prior art are deeply studied, the inventor of the application starts with the point that the residence time of the traditional ophthalmic preparation at the eyes is short and the bioavailability is low, and combines active ingredients with polymers of different in-situ gel systems, so that a surprising breakthrough is achieved, the preparation is affected by different conditions, the preparation is converted from a liquid state into a solid gel state within a period of time, the preparation has a certain viscosity, the defects of the traditional ophthalmic preparation and a post-improvement type preparation can be overcome, and besides the bioavailability of the medicine is effectively improved, the problems of poor spreadability and difficult dosage control of the traditional gel are also overcome. The temperature-sensitive gel preparation, the pH-sensitive gel preparation and the ion-sensitive gel preparation can not be lost due to objective conditions such as tear flushing after finishing phase change through environmental change, and can be adhered to eyes for at least 30min.

The invention provides a nitroimidazole in-situ gel preparation for eyes, which is used for treating eye diseases caused by Demodex. Nitroimidazoles include, but are not limited to, metronidazole, tinidazole, ornidazole, morpholine Lin Xiao; in-situ gel matrices, including but not limited to temperature sensitive polymers, pH sensitive polymers, ion sensitive polymers, are characterized in that ophthalmic formulations composed of an active ingredient, in-situ gel matrix, and other excipients, which are typically liquid under ambient conditions, after application to a specific site, the hydrogel of the composition will begin to phase change under the action of the in-situ gel matrix due to its highly porous structure, which allows for prolonged local high concentrations of drug eluting the gel after phase change in the patient's area. In some embodiments, after the nitroimidazole ophthalmic preparation is applied to the affected part of the eye, the phase of the pharmaceutical preparation starts to change under the influence of different conditions such as temperature, pH, cations and the like, and when the storage modulus is greater than the loss modulus, the preparation is changed from a liquid state to a solid state, has certain viscosity, can stay in the eye for at least 30 minutes, and overcomes the defects of poor drug spreadability, short residence time, low bioavailability and the like of the traditional ophthalmic preparation and the post-improvement type preparation.

The technical scheme of the invention can be as follows:

A. nitroimidazoles temperature sensitive ophthalmic formulations:

scheme one: a nitroimidazole temperature-sensitive ophthalmic preparation has an activity of 0.2% -5.0% of metronidazole, and the temperature-sensitive polymer comprises one or more of N-isopropyl acrylamide, poloxamer 407 and poloxamer 188, which are combined according to a certain proportion. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive, pH regulator and penetration enhancer, and are mixed with active ingredient, temperature-sensitive polymer and water according to a certain proportion. The total impurity of the metronidazole raw material is not more than 0.2%; the humectant can be propylene glycol or glycerin, and the dosage of the humectant accounts for 1% -5% of the prescription; the bacteriostatic agent is one or more of nipagin esters and benzalkonium chloride, and the total dosage is less than or equal to 1% of the prescription dosage; the bioadhesive agent can be selected from one or more of carbomer, polycarbophil, hydroxypropyl cellulose, hypromellose and sodium carboxymethyl cellulose; the preparation auxiliary materials also comprise osmotic pressure regulator. The viscosity of the preparation is 5-100Pa.s after gelation, the gelation time is 2-15 seconds, and the gelation temperature is 25-33 ℃. Can be used for treating blepharitis caused by Demodex. The drug administration is carried out through the auxiliary device, so that the drug administration uniformity can be effectively ensured, and the relative rest is ensured before the phase change is finished.

The preparation method comprises the following steps: dispersing metronidazole in humectant, adding water, dissolving thermosensitive polymer and bioadhesive in water solution of bacteriostat, mixing A, B, regulating pH regulator to 5.0-9.0, adding water, and stirring.

Scheme II: the nitroimidazole temperature-sensitive ophthalmic preparation has the activity of tinidazole, and the temperature-sensitive polymer comprises one or more of N-isopropyl acrylamide, poloxamer 407 and poloxamer 188, which are combined according to a certain proportion. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive, pH regulator and penetration enhancer, and are mixed with active ingredient, temperature-sensitive polymer and water according to a certain proportion. The viscosity of the preparation is 5-100Pa.s after gelation, the gelation time is 2-15 seconds, and the gelation temperature is 25-33 ℃. Can be used for treating blepharitis caused by Demodex. The drug administration is carried out through the auxiliary device, so that the drug administration uniformity can be effectively ensured, and the relative rest is ensured before the phase change is finished.

The preparation method comprises the following steps: dissolving tinidazole in humectant and adding a certain amount of water, dissolving temperature sensitive polymer and bioadhesive in aqueous solution of bacteriostatic agent, mixing A, B, adjusting pH regulator to 5.0-9.0, adding a certain amount of water, and stirring uniformly.

Scheme III: the nitroimidazole temperature-sensitive ophthalmic preparation has the activity of ornidazole, and the temperature-sensitive polymer is one or more of N-isopropyl acrylamide, poloxamer 407 and poloxamer 188, and is combined according to a certain proportion. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive and pH regulator, and are mixed with active ingredient, temperature-sensitive polymer and water according to a certain proportion. The viscosity of the preparation is 5-100Pa.s after gelation, the gelation time is 2-15 seconds, and the gelation temperature is 25-33 ℃. Can be used for treating blepharitis caused by Demodex. The drug administration is carried out through the auxiliary device, so that the drug administration uniformity can be effectively ensured, and the relative rest is ensured before the phase change is finished.

The preparation method comprises the following steps: dissolving ornidazole in humectant and adding water, dissolving thermosensitive polymer and bioadhesive in water solution of bacteriostat, mixing A, B, regulating pH regulator to 5.0-9.0, adding water, and stirring.

B. Nitroimidazoles pH-sensitive ophthalmic formulations:

the pH value of the normal tear of the human body is 7.4, the tolerable pH value range of the normal human eye is 5.0-9.0, and when the pH value is less than 5.0 or more than 9.0, obvious irritation can be caused, so that the secretion of the tear is increased, the medicine is quickly lost, and the cornea tissues are injured. The pH-sensitive ophthalmic preparation has a phase transition pH of 6.5-7.5, so that the pH of the prepared ophthalmic preparation needs to be controlled between 6.0-6.3 by a pH regulator. After the preparation contacts the eye to produce tears, the preparation is forced to complete the transition from liquid to gel by the pH environment of human tears.

Scheme IV: a nitroimidazole pH-sensitive ophthalmic preparation has the activity of metronidazole and the pH-sensitive polymer of chitosan. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive and pH regulator, and are mixed with active ingredient, chitosan and water according to a certain proportion to form the preparation. The viscosity of the preparation after gelation is 10-100Pa.s, and the gelation time is 2-8 seconds after contact with tear. Can be used for treating blepharitis caused by Demodex.

The preparation method comprises the following steps: a, dissolving metronidazole in a humectant and a certain amount of water, B, dissolving chitosan and a bioadhesive in a bacteriostatic agent aqueous solution, mixing A, B, adding a certain amount of water, uniformly stirring, and controlling the pH value of the preparation to be within a range of 6.0-6.3 by using a pH regulator.

Scheme five: a nitroimidazole pH-sensitive ophthalmic preparation has the activity of tinidazole and a pH-sensitive polymer of chitosan. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive and pH regulator, and are mixed with active ingredient, chitosan and water according to a certain proportion to form the preparation. The viscosity of the preparation after gelation is 10-100Pa.s, and the gelation time is 2-8 seconds after contact with tear. Can be used for treating blepharitis caused by Demodex.

The preparation method comprises the following steps: a, dissolving tinidazole in a humectant and a certain amount of water, B, dissolving chitosan and a biological adhesive in a bacteriostatic agent aqueous solution, mixing A, B, adding a certain amount of water, uniformly stirring, and controlling the pH value of the preparation to be within a range of 6.0-6.3 by using a pH regulator.

Scheme six: a nitroimidazole pH-sensitive ophthalmic preparation has the activity of ornidazole and the pH-sensitive polymer of chitosan. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive and pH regulator, and are mixed with active ingredient, chitosan and water according to a certain proportion to form the preparation. The viscosity of the preparation after gelation is 10-100Pa.s, and the gelation time is 2-8 seconds after contact with tear. Can be used for treating blepharitis caused by Demodex.

The preparation method comprises the following steps: ornidazole is dissolved in humectant and a certain amount of water, chitosan and bioadhesive are dissolved in bacteriostatic agent water solution, A, B is mixed and added with a certain amount of water, and the mixture is stirred uniformly, and the pH value of the preparation is controlled within the range of 6.0-6.3 by using pH regulator.

C. Nitroimidazole ion-sensitive ophthalmic formulation:

ion-sensitive matrix means that it is capable of responding to cations in the external environment, thereby completing a phase change. Deacetylated gellan gum is particularly sensitive to calcium and magnesium ions, which are present in tears. After the preparation contacts the eyes to generate tears, the preparation can be converted from liquid state to gel state through the cationic environment brought by human tears.

Scheme seven: a nitroimidazole ion-sensitive ophthalmic preparation has the activity of metronidazole and the ion-sensitive polymer of deacetylated gellan gum. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive and pH regulator, and are mixed with active ingredient, deacetylated gellan gum and water according to a certain proportion. The viscosity of the preparation after gelation is 5-100Pa.s, and the gelation time is 2-12 seconds after contact with tear. Can be used for treating blepharitis caused by Demodex.

The preparation method comprises the following steps: dissolving metronidazole in humectant and water, dissolving deacetylated gellan gum and bioadhesive in water solution of bacteriostat, mixing A, B, regulating pH regulator to 5.0-9.0, adding water, and stirring.

Scheme eight: a nitroimidazole ion-sensitive ophthalmic preparation has the activity of tinidazole and the ion-sensitive polymer of deacetylated gellan gum. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive and pH regulator, and are mixed with active ingredient, deacetylated gellan gum and water according to a certain proportion. The viscosity of the preparation after gelation is 5-100Pa.s, and the gelation time is 2-12 seconds after contact with tear. Can be used for treating blepharitis caused by Demodex.

The preparation method comprises the following steps: dissolving tinidazole in humectant and adding a certain amount of water, dissolving deacetylated gellan gum and bioadhesive in antibacterial agent water solution, mixing A, B, adjusting pH regulator to 5.0-9.0, adding a certain amount of water, and stirring uniformly.

Scheme nine: a nitroimidazole ion-sensitive ophthalmic preparation has the activity of ornidazole and the ion-sensitive polymer of deacetylated gellan gum. The other auxiliary materials are selected from one or more of humectant, antibacterial agent, biological adhesive and pH regulator, and are mixed with active ingredient, deacetylated gellan gum and water according to a certain proportion. The viscosity of the preparation after gelation is 5-100Pa.s, and the gelation time is 2-12 seconds after contact with tear. Can be used for treating blepharitis caused by Demodex.

The preparation method comprises the following steps: dissolving ornidazole in humectant and adding water, dissolving deacetylated gellan gum and bioadhesive in water solution of bacteriostat, mixing A, B, regulating pH regulator to 5.0-9.0, adding water, and stirring.

In certain embodiments, optional in-situ gel matrices include, but are not limited to, polyethers (e.g., polyethylene oxide, polypropylene oxide, polyethylene oxide-polypropylene oxide copolymers), poloxamers, celluloses, and hemicelluloses (e.g., methylcellulose, hydroxypropyl methylcellulose, ethyl (hydroxyethyl) cellulose, xyloglucan), acetates, phthalates, latexes, polyacrylates (e.g., polyacrylic acid), N-alkylacrylamides (e.g., poly (N-isopropylacrylamide, amine)), hyaluronic acid, chitosan, dextran, and gellan gum, and derivatives thereof; in some embodiments, the polymer or copolymer comprises polyethylene oxide or polypropylene oxide; in some embodiments, the copolymer comprises a polyethylene/polypropylene copolymer or a polyethylene/polypropylene block copolymer; in some embodiments, the copolymer comprises a poloxamer; in some embodiments, the copolymer comprises poloxamer 407, poloxamer 188, poloxamer sarin, poloxamer 124, poloxamer 237, or poloxamer 338; in some embodiments, the copolymer comprises poloxamer 407; in certain embodiments, the polymer or copolymer comprises a phosphate monomer; in certain embodiments, the copolymer comprises poloxamer and phosphate ester monomers.

In certain embodiments, the bacteriostatic agent is any one or more of thimerosal, benzalkonium bromide, benzalkonium chloride, parabens; in certain embodiments, the pH adjuster is any one or more of phosphoric acid, hydrochloric acid, sodium hydroxide; in certain embodiments, the humectant is any one or more of glycerin, lactic acid, 1, 2-hexanediol, octanediol, allantoin, sodium hyaluronate; in certain embodiments, the bioadhesive is any one or more of carbomer, gelatin, hydrolyzed collagen, cellulose, xanthan gum, sodium alginate; the method comprises the steps of carrying out a first treatment on the surface of the In certain embodiments, the osmolality adjusting agent is any one or more of sodium chloride, glucose, boric acid.

Temperature sensitive ophthalmic formulations the composition is a liquid below the phase transition temperature. In some embodiments, the phase transition temperature is at or below eye temperature (e.g., about 33 ℃). Thus, the composition forms a gel when applied to the subject's eye, for example, when the composition contacts a biological surface. In some embodiments, the phase transition temperature is from about 0 ℃ to about 33 ℃, from about 10 ℃ to about 33 ℃, from about 15 ℃ to about 33 ℃, from about 20 ℃ to about 33 ℃, from about 25 ℃ to about 33 ℃, from about 30 ℃ to about 33 ℃, or from about 31 ℃ to about 33 ℃; in some embodiments, the phase transition temperature is from about 20 ℃ to about 37 ℃; in some embodiments, the phase transition temperature is from about 25 ℃ to about 35 ℃; in some embodiments, the phase transition temperature is about 20 ℃ to 25 ℃, about 25 ℃ to about 30 ℃, about 30 ℃ to about 35 ℃, or about 35 ℃ to about 40 ℃.

In certain embodiments, the compositions are useful for treating diseases; in some embodiments, the composition is useful for treating an infectious disease; in some embodiments, the composition is useful for treating an ocular disease; in some embodiments, the composition may be used to treat blepharitis. In some embodiments, the composition is useful for treating demodex blepharitis.

The phase transition temperature of the formulation is one factor in determining whether the composition is suitable. The temperature at which the storage modulus exceeds the loss modulus is considered the gelation temperature. The gelling temperature of the compositions herein may be below or above 33 ℃, but is preferably below 33 ℃, to accelerate the gelling of the composition immediately after administration, particularly after exposure of the matrix forming agent to the eye. The timing of the sol-gel transition will affect the ease of application. In certain embodiments, the composition is internal gelled at the edge of the applied eyelid for about 3 seconds, about 5 seconds, about 10 seconds, about 20 seconds, about 30 seconds, about 1 minute; in some embodiments, the composition gels from about 1 second to about 20 seconds after application. In certain embodiments, the composition is stored refrigerated (e.g., refrigerated at about 5 ℃) prior to administration. Cold storage may be useful for compositions having a gelation temperature below room temperature to prevent gelation prior to application or during handling.

Drawings

Fig. 1 is an eye structure diagram.

Detailed Description

In order that those skilled in the art will better understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in which it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, shall fall within the scope of the invention.

Example 1

Metronidazole temperature-sensitive ophthalmic preparation prescription and preparation process thereof

Metronidazole: 0.75%

Poloxamer 407:18%

Poloxamer 188:3.2%

Glycerol: 5%

Polycarbophil: 0.5%

Benzalkonium chloride: 0.05%

Sodium hydroxide adjusts the pH value to be: 6.5 to 8.0

Purified water was about: 72.50%

The preparation method comprises the following steps: (1) dissolving metronidazole in a mixed solution of glycerol and part of water; (2) poloxamer 407, poloxamer 188 and polycarbophil are respectively swelled in benzalkonium chloride aqueous solution; (3) and (3) uniformly mixing the components (1) and (2), regulating the pH value to 7.4 by using a sodium hydroxide solution, adding the rest water, and uniformly stirring to obtain the water-soluble emulsion.

Experimental results: the gelation temperature of the obtained eye metronidazole temperature-sensitive gel sample is 28-30 ℃, the viscosity after gelation is about 15Pa s, and the gelation time is about 5 seconds.

Example 2

Metronidazole: 2%

Poloxamer 407:32%

Poloxamer 188:6%

Propylene glycol: 9%

Hypromellose: 4%

Ethyl hydroxy benzoate: 0.1%

Sodium hydroxide adjusts the pH value to be: 6.5 to 8.0

Purified water was about: 46.50%

The preparation method comprises the following steps: (1) dissolving metronidazole in a mixed solution of propylene glycol and part of water; (2) poloxamer 407, poloxamer 188 and hypromellose are respectively swelled in an aqueous solution of ethyl hydroxybenzoate; (3) and (3) uniformly mixing the components (1) and (2), regulating the pH value to 7.4 by using a sodium hydroxide solution, adding the rest water, and uniformly stirring to obtain the water-soluble emulsion.

Experimental results: the gelation temperature of the obtained metronidazole temperature-sensitive gel sample for eyes is 29-33 ℃, the viscosity after gelation is about 20Pa s, and the gelation time is about 10-12 seconds.

Example 3

The test and analysis method of the pharmaceutical preparation were carried out by using the ophthalmic preparation prepared in example 1.

A. System applicability

In a liquid chromatogram of the reference solution, the theoretical plate number of the metronidazole peak is 8000; the degree of separation between the metronidazole peak and its impurity I peak was 4.1; in the sensitivity solution liquid chromatograph, the signal to noise ratio of the main peak is more than 10, and the three data structures indicate that the system has good applicability.

B. Specialization of

The blank solvent/negative test sample has no interference to the detection of the metronidazole and the metronidazole impurity I, which shows that the method has good specificity.

C. Durability of

When the chromatographic column, the column temperature and the flow rate change slightly, the requirements of the system applicability can be met, the influence on the detection result is small, and the durability is good.

D. Stability of

The sample solution is placed for 48 hours at room temperature for continuous sampling for 30 needles, the absolute difference between the impurity I and the total impurity content is less than 0.1 percent, and the absolute difference between the total impurity content is less than 0.1 percent, which indicates that the sample solution is stable when placed for 48 hours at room temperature.

E. Repeatability of

The average content of impurity I in 9 parts of marked test sample solution is 0.14%, and the content RSD=1.0% < 10%; total miscellaneous rsd=3.0% < 5.0%, good repeatability.

F. Limit of detection and limit of quantification

Metronidazole was detected at a limit of 0.2101ng and quantified at a limit of 0.4204ng. The detection limit of the impurity I is 0.1564ng, and the quantitative limit is 0.6315ng.

G. Linearity and range

Metronidazole has a linear relationship between concentration and peak area in the range of 0.04204-0.8061 μg/ml, the linear equation is y=51.78x+1.4, the correlation coefficient r=0.9999 > 0.998, and the sum of squares of the residuals is 0.01. The impurity I is in the range of 0.03101-4.672 mug/ml, the concentration and the peak area are in linear relation, the linear equation is y=51.41x+3.483, the correlation coefficient r=0.9997 > 0.998, and the sum of squares of residual errors is 0.01.

Example 4

The content measurement and detection method of the pharmaceutical preparation was verified by the ophthalmic preparation prepared in example 1.

A. System applicability

The separation degree between the metronidazole peak and the metronidazole impurity I peak is 4.0 and is more than 2.0. The system has good applicability.

B. Specialization of

And taking a control substance solution of the content measurement item, a test substance solution and a negative sample solution of the metronidazole, and measuring by the same method, wherein the result shows that the retention time of a main peak in the test substance solution is the same as that of the main peak of the metronidazole control substance solution, and the negative sample has no same characteristic peak, so that the negative sample has no interference.

C. Linearity and range

Under the condition of the intrinsic color spectrum, the concentration of the metronidazole is in a range of 0.06172mg/ml-0.16452mg/ml, the linear relation between the concentration and the peak area is shown, the linear equation is y=50665x+103.12, the correlation coefficient r=0.9997, and the sum of squares of residual errors is 0.000004.

D. Accuracy of

Samples were treated and recovery was measured at three concentrations, 80%, 100% and 120%, respectively, resulting in an average recovery of 99.17% and rsd=0.2%.

E. Precision of instrument

The control solution under the content measurement item is continuously measured for 6 times, the peak area and the retention time RSD are respectively 0.2% and 0.1%, and the precision is good.

F. Repeatability of

Taking 9 parts of sample solution with the average content of 101.5% and the content RSD=0.2% < 4.0% under the content measurement; the repeatability is good.

G. Intermediate precision

Two technicians prepare 12 parts of repeated test sample solution/intermediate precision solution, and the average content of the metronidazole is 101.4 percent, and the content RSD=0.3% < 2.0 percent measured by different instruments; meets the requirements. Another technician prepares 6 parts of intermediate precision solution, the average content of the metronidazole is 101.6%, and the content RSD=0.3% < 2.0%; meets the requirements.

H. Solution stability

Taking a reference substance solution and a test substance solution under the content measurement item, placing under the room temperature environment, and measuring peak areas of the reference substance solution and the test substance solution at 0, 4, 8, 12, 18, 24, 48 and 72 hours respectively, wherein RSD is 0.3% and 0.2% respectively, and the solution stability is good.

I. Durability of

The durability is better, and when the chromatographic column, the column temperature and the flow velocity are slightly changed, the requirements of the system applicability can be met, and the influence on the measurement result is smaller.

The examples described in this application are provided to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting the scope thereof.

Claims (10)

1. A nitroimidazole temperature-sensitive ophthalmic formulation, characterized by:

A. comprises one or more nitroimidazole drugs as active ingredients, wherein the nitroimidazole drugs comprise metronidazole, tinidazole, ornidazole and morpholine Lin Xiao;

B. comprises one or more combinations of temperature-sensitive polymers as auxiliary materials, and is characterized in that: formulations composed of temperature sensitive polymers have a lower storage modulus than loss modulus at below 25 ℃; the storage modulus is higher than the loss modulus at 33 ℃ and above; the temperature range of storage modulus equal to loss modulus is between 25 ℃ and 33 ℃.

2. A nitroimidazole temperature-sensitive ophthalmic formulation as claimed in claim 1, wherein the temperature-sensitive polymer types include: polyethylene oxide, polypropylene oxide, polyethylene oxide-polypropylene oxide copolymers, poloxamer 407, poloxamer 188, poloxamer 124, poloxamer 237 or poloxamer 338, poloxamer 407, poloxamer salad amine, methylcellulose, hydroxypropyl methylcellulose, ethyl (hydroxyethyl) cellulose, xyloglucan, acetate, phthalate, latex, polyacrylic acid, N-alkyl acrylamide, hyaluronic acid, chitosan, dextran and gellan gum, and derivatives thereof.

3. The nitroimidazole temperature-sensitive ophthalmic preparation according to claim 1, wherein the active ingredient is metronidazole and the content of the active ingredient is 0.2% -5.0%.

4. The nitroimidazole temperature-sensitive ophthalmic formulation of claim 1, wherein the formulation excipients further comprise: one or more of humectant, antibacterial agent, bioadhesive, pH regulator, osmotic pressure regulator and permeation promoter are selected.

5. The nitroimidazole temperature-sensitive ophthalmic preparation according to claim 1-4, wherein the semisolid gel formed by the preparation under the influence of temperature when the preparation acts on the eyelid has a viscosity of 5-100 Pa.s.

6. The nitroimidazole temperature-sensitive ophthalmic preparation according to claim 1-5, wherein the phase transition temperature of the preparation is 25-33 ℃.

7. The nitroimidazole temperature-sensitive ophthalmic preparation according to claim 1-6, wherein the conversion time of the preparation from a liquid state to a solid state gel is 2-20 seconds.

8. The nitroimidazole temperature-sensitive ophthalmic preparation according to claim 1-7, wherein the total impurities of the active ingredient metronidazole raw material are not more than 0.2%; the content of impurity I is not more than 0.1%.

9. A nitroimidazole temperature-sensitive ophthalmic preparation, which is characterized by being used for treating eye diseases caused by demodex.

10. A nitroimidazole temperature-sensitive ophthalmic formulation as claimed in claim 9 for the treatment of blepharitis caused by demodex, in particular including anterior blepharitis caused by demodex folliculorum and meibomian dysfunction caused by demodex sebaceous.