CN117883380B

CN117883380B - A transdermal emulsion for treating dry eye

Info

Publication number: CN117883380B
Application number: CN202311737599.2A
Authority: CN
Inventors: 接英; 田磊
Original assignee: Beijing Tongren Hospital
Current assignee: Beijing Tongren Hospital
Priority date: 2023-12-18
Filing date: 2023-12-18
Publication date: 2025-04-01
Anticipated expiration: 2043-12-18
Also published as: CN117883380A

Abstract

The present invention relates to a transdermal emulsion for treating dry eye, which comprises low-concentration menthol as the only active ingredient, an oil phase matrix, an emulsifier and an aqueous phase; the mass concentration of menthol in the emulsion composition is 1.5%-7%, and the mass of the oil phase matrix in the emulsion composition does not exceed 30%. The mass concentration of menthol in the emulsion of the present invention is 1.5%-7%, and the content of the oil phase matrix does not exceed 30%. It can effectively promote the tear secretion of dry eye patients to treat dry eye while avoiding skin irritation and greasy feeling, improve the patient's use experience, maintain the efficacy of long-term use, and is more friendly to users with oily skin, avoiding the strong greasy feeling and acne or local dermatitis caused by applying oily liquid preparations to the skin around the eyes or the nose (the area around the nose is a grease-rich area).

Description

A Chinese medicinal composition for treating xerophthalmia is administered transdermally

Technical Field

The invention relates to the technical field of dry eye disease prevention and treatment medicines, in particular to a percutaneous administration emulsion for treating dry eye disease.

Background

The pathogenesis of dry eye is quite complex and is not fully understood so far no good treatment for dry eye has been known. Dry eye causes are various, and the effects of environmental factors, ocular inflammation, systemic diseases, electronic equipment, air conditioning facilities and the like can cause dry eye. The dry eye can cause a plurality of ocular discomfort, such as dry eyes, itching and the like, and the serious development of the dry eye of a part of patients can even lead to a certain degree of damage to corneal epithelium, a great deal of secretion of eyes, conjunctival congestion, serious pathological changes of eyes and the like, and the vision is seriously affected for a long time. The tears contain various lysozyme, mucopolysaccharide, immune protein and other components, can prevent eye infection, moisten and lubricate the eye surface, and has important protection and nutrition effects on eyes. The most prominent symptom of dry eye is a significant decrease in tear secretion, which causes ocular discomfort. Thus, the treatment of dry eye should be initiated by protection of tears and promotion of tear secretion.

Currently, a variety of eye drops are available on the market for the treatment of dry eye, such as sodium hyaluronate eye drops, cyclosporin a eye drops, and the like. However, the cause of dry eye is complicated, and the mechanism is not completely clear, but the eye drops for treating dry eye on the market can make patients feel comfortable in a short time after eye drop administration, and reduce symptoms such as eye pain, but the effect of relieving the dry eye symptoms is often short in duration, and the repeated use of the eye drops for a long time can cause complications such as corneal toxicity and drug-induced dry eye, and further aggravate the dry eye symptoms of the patients, so that a vicious circle is formed (Malik Kahoo, cullin T.Vogelson, usha Sivaprasad, david A.Ammar. Different artificial tears have toxicity effects on human keratoconjunctival epithelial cells cultured in vitro compared with [ J ]. J.J.J.J. 2012,48 (2): 170-172.). In addition, the use of an ophthalmic solution such as artificial tears for a long period of time affects the functions of the eyes themselves, and the eye relies on the ophthalmic solution to reduce tear secretion, and tear secretion function of lacrimal glands gradually deteriorates, thereby forming a further vicious circle (Lv. Commonly used ophthalmic solutions are carefully "dry eye" [ J ]. Family science and new health, 2010 (5): 27).

The drug OC-01 (valicalan, CAS: 249296-44-4) nasal spray related to the invention patent application CN202010304809 can treat xerophthalmia as well, and the drug is marketed abroad and is currently in clinical test in China. The medicine is administered through nasal cavity, and can stimulate trigeminal nerve and then stimulate lacrimal secretion. However, the medicine has strong nasal cavity irritation, and is counted to cause sneeze of 82% of patients, and adverse reactions such as nasal cavity irritation, cough, sore throat and the like of 5-16% of patients (data are derived from the drug specifications of the medicine marketed in the United states, and the drug trade name is Tyrvaya).

It is also reported in literature that oral administration of bromhexine, pilocarpine hydrochloride, neostigmine and the like can promote secretion of tears of partial patients, but the actual curative effect is still uncertain. In addition, patients with xerophthalmia need to take medicines for a long time, and the defect of oral medicines is that toxic and side effects cannot be ignored. For example, pilocarpine, an oral drug, is clearly shown in the drug specification to cause nausea, vomiting, abdominal pain, diarrhea, salivation, miosis, rapid and slow heart beat, microcrack cramp, excessive mucous secretion and the like, which can cause dyspnea, visual discomfort caused by taking the pilocarpine, and reduce night driving safety, and if patients continuously sweat while taking the pilocarpine and cannot supplement enough beverage, the patients must go to the hospital to check treatment to prevent collapse. This is also the main reason why oral drugs are rarely used in the clinical treatment of dry eye.

Menthol is also called menthol, hexahydrothymol, 5-methyl-2-isopropyl cyclohexanol, and is the main ingredient in peppermint and peppermint essential oils. In nature, menthol exists mainly in the form of L in essential oils of mint-like plants. Modern medicine proves that the mint has excitation effect, can expand skin capillary, dispel wind-heat, can be used for treating cold wind-heat, has good throat effect, can be used for treating wind-heat pharyngalgia and cough, has penetrating effect, and can help measles to penetrate. Herba Menthae also has analgesic, antipruritic, antibacterial, and insecticidal effects. The fresh mint leaves are rubbed and then stuck on the temple, so that the vasodilating headache can be treated. In addition, peppermint can treat rhinitis, resist sepsis, treat skin diseases, etc., and is widely used as a spice in toothpaste, chewing gum, wine, beverages, cosmetics, etc.

U.S. patent application No. US202117511238A (title of invention: TREATMENT FOR DRY EYES) discloses a composition of balm mixture (pasty mixture) containing Menthoxypropanediol, menthonediol, menthol and Caffeine, applied to periocular skin for the treatment of dry eye. However, the active ingredient mixture disclosed in the patent of the invention has a complex composition, the Menthoxypropanediol concentration is 30-40%, the Menthonediol concentration is 30-40%, the menthol concentration is 30-40%, and the Caffeine concentration is 0.01-0.25%. . The content of the menthol and related mint components in the medicine prescription of the invention is up to 20-60 percent, and the menthol and related mint components are smeared on the skin (adjacent the lower eyelid) near the lower eyelid, and the menthol and related mint components are extremely easy to cause strong skin irritation due to the excessive concentration, including skin redness, burning sensation, itching and other discomforts of the smeared part. Since the periocular skin itself is thinner than the other parts, the stratum corneum is also thinner, leading to the periocular skin being sensitive and fragile, whereas menthol and its analogues are more irritating to the skin and more irritating at higher concentrations. The composition is solid wax, is smeared on the skin, active ingredients such as menthol and the like are blocked by the wax and cannot be completely absorbed through the skin quickly, but the menthol and the like with high concentration stay near the eyelid for a long time and have strong volatility, so that the volatile is irritated to eyes, the eyes of a patient are extremely uncomfortable to smoke, the use experience of a user is poor, and most importantly, the effect of promoting tear secretion is not ideal in practice even though the menthol and the like with the concentration of up to 20-60% are used. Thus, the compositions disclosed in this patent application have very low potency and strong irritating side effects can also lead to low patient compliance.

Heretofore, the applicant has filed a patent application (application number 202311621663.0) to the national bureau of patent for a transdermal drug delivery composition for treating dry eye, which is prepared with menthol at a low concentration as the only active ingredient, and which is mainly an oily liquid preparation dispersed in vegetable oil, and which can be sprayed or smeared, and which has an oil content of up to 90 to 97%. Experiments prove that the composition can effectively promote the self tear secretion of a xerophthalmia patient, treat xerophthalmia without causing skin irritation, but the use experience of the composition is poor in the feedback of the patient in the further trial process, and the specific reasons are that the xerophthalmia is a chronic disease, long-time application is needed, the greasy feeling of a liquid preparation with high oil content is very strong, the skin feel experience of the patient is poor after continuous repeated application, the skin feel experience of the patient is more obvious especially for oily skin, after continuous repeated application, the greasy feeling of the face and the easy acne and local skin inflammation are generated after the grease secreted by human bodies around the nasal wings are mixed, the oil covers the pores before the application part, and the curative effect is reduced after continuous repeated continuous application. In order to improve the use experience and the curative effect of long-term use, an improvement scheme is required to be provided.

The inventor researches find that the problem of the oily liquid preparation can be well solved by preparing menthol into emulsion. Furthermore, a lower concentration of menthol in the emulsion achieves the desired effect of promoting tear secretion.

Disclosure of Invention

First, the technical problem to be solved

In view of the above-mentioned shortcomings and disadvantages of the prior art, the present invention provides a transdermal administration emulsion for treating dry eye, wherein the mass concentration of menthol in the emulsion is 1.5% -7%, and the content of oil phase matrix is not more than 30%, so that the self-tear secretion of a dry eye patient can be effectively promoted to treat dry eye, and meanwhile, skin irritation and greasy feeling can be avoided, the use experience of the patient can be improved, the long-term use curative effect can be maintained, and the transdermal administration emulsion is more friendly to oily skin users.

(II) technical scheme

The invention comprises the following technical scheme:

In a first aspect, the present invention provides an emulsion for transdermal administration for treating dry eye, comprising menthol at a low concentration of 1.5% to 7% by mass of the emulsion, an oil phase matrix, an emulsifier, and an aqueous phase as the sole active ingredients, the oil phase matrix constituting no more than 30% by mass of the emulsion, preferably, the menthol constituting 2% to 6% by mass of the emulsion, more preferably, 3% to 5% by mass of the emulsion.

When the oil phase matrix accounts for not more than 30% of the emulsion, the cream can greatly reduce the greasy feeling of being smeared on the face, avoid skin irritation and greasy feeling when treating xerophthalmia, promote the use experience of patients, and is more friendly to oily skin users in particular.

According to a preferred embodiment of the invention, the menthol is natural menthol or synthetic menthol.

According to a preferred embodiment of the present invention, the oil phase base is selected from one or more of tea oil, olive oil, soybean oil, coconut oil, sunflower oil, corn oil, avocado oil, shea butter, grape seed oil, almond oil, castor oil, jojoba oil, lanolin, beeswax, medium chain triglycerides, isopropyl palmitate, safflower oil, stearyl alcohol, stearic acid, cetyl alcohol, cholesterol, silicone oil, dimethicone, liquid paraffin, light liquid paraffin.

According to a preferred embodiment of the present invention, the emulsifier is at least one selected from lecithin, polyoxyethylene (35) castor oil, polyoxyethylene (40) hydrogenated castor oil, tween 80, span 80, poloxamer 188, poloxamer 407, sodium stearate, potassium stearate, sodium dodecyl sulfate, sodium cetyl sulfate, cetyl alcohol, polyoxyethylene 40 stearate, polyoxyethylene lauryl ether, sucrose fatty acid ester.

According to a preferred embodiment of the invention, the emulsion consists of 1.5% -7% menthol, 3% -25% oil phase matrix, 0.05% -3% emulsifier and the balance aqueous phase. Preferably, the menthol is present in a mass concentration of 2% to 6%, more preferably 3% to 5%. Among them, the oil phase matrix is preferably 5 to 20%, more preferably 5 to 15%. The lower the content of the oil phase matrix, the cleaner and thinner the emulsion, and the cleaner and more comfortable the emulsion is after being applied on the facial skin (the skin around the eyelid or the skin at the nasal wings and nasal labial sulcus on both sides).

Preferably, the emulsion is prepared by dissolving menthol in an oil phase matrix to obtain an oil phase, dissolving a water-soluble emulsifier in water if the emulsifier is a water-soluble emulsifier, dissolving an oil-soluble emulsifier in the oil phase matrix if the emulsifier is an oil-soluble emulsifier, mixing the water phase and the oil phase, and rapidly stirring or homogenizing by a homogenizer or shearing uniformly by a shearing machine to obtain the emulsion.

According to a preferred embodiment of the present invention, the aqueous phase is water or an aqueous solution in which a water-soluble auxiliary agent is dissolved, for example, at least one of a water-soluble essence, a preservative (such as propyl paraben), and a stabilizer (such as disodium EDTA).

In a second aspect, the present invention relates to the use of the emulsion described above for the preparation of a medicament for the treatment of dry eye.

Preferably, the application comprises applying the emulsion to periocular skin or to skin at the sides of the nasolabial folds and nasolabial folds. Periocular skin includes skin at the upper eyelid and skin at the lower eyelid. More preferably, the emulsion is applied to the skin at the lower eyelid and the skin at the nasal wings and nasolabial folds on both sides.

The inventor finds in the research that only low-concentration menthol is used for the skin around eyes (the skin at the upper eyelid and the skin at the lower eyelid) or the skin at the nasal wings and the nasal labial sulcus at the two sides, can obviously promote tear secretion of xerophthalmia patients, can effectively treat xerophthalmia, has no irritation to the skin and has huge clinical application potential. Further, the inventor finds that the low-concentration menthol is prepared into emulsion by pharmaceutically acceptable auxiliary materials, compared with an oily liquid preparation, the emulsion is more beneficial to transdermal absorption of menthol, and the curative effect and the onset speed of the low-concentration menthol for treating xerophthalmia are improved. Subsequent researches further find that the emulsion containing the menthol with low concentration is more beneficial to transdermal absorption of menthol, further improves the efficacy of the menthol with low concentration for treating xerophthalmia (the menthol concentration required by the same curative effect is lower than that of the oily liquid preparation), solves the problem that the curative effect is reduced because the skin of a medicine application part is covered with more oil layers after the oily liquid preparation is continuously used for multiple times, and effectively solves the problem of greasy skin feel generated by the continuous multiple times of use of a user, improves the experience of the user, and is more friendly to oily skin patients.

(III) beneficial effects

The invention provides a percutaneous administration emulsion which takes menthol with low concentration as the only active ingredient, can effectively promote the secretion of tears of patients, has obvious effect of relieving dry eye symptoms of dry eye patients, and does not irritate facial skin and generate the problem that eyes are difficult to open because of low menthol content.

The mass concentration of menthol in the emulsion is 1.5% -7%, the content of the oil phase matrix is not more than 30%, the emulsion can effectively promote the secretion of the tear of a xerophthalmia patient to treat xerophthalmia, simultaneously avoid skin irritation and greasy feeling, promote the use experience of the patient, maintain the curative effect of long-term use, and especially be more friendly to oily skin users, and avoid the problems of strong greasy feeling, acne or local dermatitis and the like caused by the fact that the oily liquid preparation is smeared around nosewings (the fat-rich area around the nosewings).

In addition, compared with the oily liquid preparation with the same curative effect, the emulsion provided by the invention has the advantages that the required menthol concentration is lower, the problem that the continuous repeated use curative effect is reduced due to the fact that the oil coating layer is arranged on the application part after the oily liquid preparation is continuously used for multiple times is solved, and the user experience is better.

Drawings

Fig. 1 is a photograph of skin irritation of experimental drug group 2 in the experiment of example 2.

Fig. 2 is a photograph of skin irritation of experimental drug group 5 in the experiment of example 2.

Fig. 3 is a photograph of skin irritation of experimental drug group 8 in the experiment of example 2.

Fig. 4 is a photograph of skin irritation of the oil-soluble control drug 3 group in the experiment of example 2.

Fig. 5 is a photograph of skin irritation of the oil-soluble control drug group 7 in the experiment of example 2.

Fig. 6 is a photograph of skin irritation of the oil-soluble control drug group 8 in the experiment of example 2.

FIG. 7 is a photograph of skin irritation of solid control drug group 1 in the experiment of example 2.

Fig. 8 is a photograph of skin irritation of the solid-state control drug group 2 in the experiment of example 2.

Fig. 9 is a photograph of an alignment of the greasy effect of different formulations on skin in the experiment of example 5.

FIG. 10 is a corneal plot of the sodium fluorescein staining of each group in the experiment of example 9.

Figure 11 is the average scoring results for each group of sodium fluorescein staining in the experiment of example 9.

FIG. 12 shows the measurement results of the secretion amount of tears in each group in the experiment of example 9.

FIG. 13 shows the measurement results of the secretion amount of tears in each group in the experiment of example 10.

FIG. 14 is a graph of staining of various groups of sodium corneal fluorescein in the experiment of example 11.

Figure 15 is the average scoring results for each group of sodium fluorescein staining in the experiment of example 11.

FIG. 16 shows the results of measurement of tear secretion in each group in the experiment of example 11.

Detailed Description

The invention will be better explained by the following detailed description of the embodiments with reference to the drawings.

Example 1 preparation of emulsion with menthol as the sole active ingredient

Specifically, the present example provides specific examples of menthol-based dermatological pharmaceutical compositions, such as the following specific example medicaments and their preparation as follows:

Experimental drug 1:0.1 g menthol was dissolved in 0.9g olive oil, 0.2g tween 80 was dissolved in 8.8ml water, then 8.8ml water with tween 80 dissolved was slowly added to olive oil under rapid stirring to give 1% emulsion (oil 9%).

Experimental drug 2.15 g menthol was dissolved in 0.85g olive oil, 0.2g tween 80 was dissolved in 8.8ml water, then 8.8ml water with tween 80 dissolved was slowly added to olive oil under rapid stirring to get 1.5% emulsion (oil content 8.5%).

Experimental drug 3.2 g menthol was dissolved in 0.8g olive oil, 0.2g tween 80 was dissolved in 8.8ml water, then 8.8ml water with tween 80 dissolved was slowly added to olive oil under rapid stirring to give 2% emulsion (oil content 8%).

Experimental drug 4:0.3 g menthol was dissolved in 1.5g olive oil, 0.2g tween 80 was dissolved in 8ml water, then 8ml water with tween 80 dissolved was slowly added to olive oil under rapid stirring to obtain a 3% emulsion (15% oil).

Experiment drug 5.4 g menthol was dissolved in 2g olive oil, 0.2g tween 80 was dissolved in 7.4ml water, then 7.4ml water with tween 80 dissolved was slowly added to olive oil under rapid stirring to give a 4% emulsion (oil 20%).

Experimental drug 6:0.5 g menthol was dissolved in 2.5g olive oil, 0.2g tween 80 was dissolved in 6.8ml water, then 6.8ml water with tween 80 dissolved was slowly added to olive oil under rapid stirring to give a 5% emulsion (oil 25%).

Experimental drug 7.6 g menthol was dissolved in 2.5g olive oil, 0.2g tween 80 was dissolved in 6.7ml water, then 6.7ml water with tween 80 dissolved was slowly added to olive oil under rapid stirring to obtain a 6% emulsion (oil 25%).

Experimental drug 8.7 g menthol was dissolved in 2.5g tea oil, 0.06g poloxamer 188 was dissolved in 6.74ml water, then 6.74ml water with tween 80 dissolved therein was slowly added to tea oil with rapid stirring to give a 7% emulsion (oil 25%).

Experimental drug 9:0.3 g menthol was dissolved in 1.5g soybean oil, 0.2g tween 80 was dissolved in 8ml water, then 8ml water with tween 80 dissolved was slowly added to soybean oil with rapid stirring to give a 3% emulsion (15% oil).

Experimental drug 10.3 g menthol was dissolved in 1.5g castor oil, 0.2g tween 80 was dissolved in 8ml water, then 8ml water with tween 80 dissolved was slowly added to castor oil with rapid stirring to give a 3% emulsion (15% oil).

Experiment drug 11 1.5g of beeswax was melted under 70 ℃ heating, then 0.3 g of menthol was dissolved in beeswax, 0.2g of tween 80 was dissolved in 8ml of water and heated to 70 ℃, then the 8ml of water was slowly added to beeswax under rapid stirring to give a 3% emulsion (15% containing beeswax).

Experimental drug 12:0.3 g menthol was dissolved in 1.5g medium chain triglyceride, 0.2g tween 80 was dissolved in 8ml water, and then the 8ml water was slowly added to the medium chain triglyceride with rapid stirring to give a 3% emulsion (15% oil).

Experiment drug 13.3 g menthol was dissolved in 1.5g isopropyl palmitate, 0.2g tween 80 was dissolved in 8ml water, and then the 8ml water was slowly added to isopropyl palmitate with rapid stirring to give a 3% emulsion (15% oil).

Experimental drug 14:0.6 g menthol was dissolved in 1.5g castor oil, 0.1g polyoxyethylene (40) castor oil was dissolved in 7.8ml water, and then 7.8ml water was slowly added to the castor oil with rapid stirring to give a 6% emulsion (15% oil).

Experimental drug 15:0.3 g menthol was dissolved in 1.5g castor oil, 0.1g polyoxyethylene (40) castor oil was dissolved in 8.1ml water, and then the 8.1ml water was slowly added to the castor oil with rapid stirring to give a 3% emulsion (15% oil).

Experimental drug 16:0.6 g menthol was dissolved in 1.5g tea oil, 0.1g polyoxyethylene (35) castor oil was dissolved in 7.8ml water, and then 7.8ml water was slowly added to tea oil with rapid stirring to give a 6% emulsion (15% oil).

Experimental drug 17.6 g menthol was dissolved in 1.5g tea oil, 0.1g poloxamer 188 was dissolved in 7.8ml water, and then 7.8ml water was slowly added to tea oil with rapid stirring to give a 6% emulsion (15% oil).

Experimental drug 18:0.3 g menthol was dissolved in 1.5g tea oil, 0.1g polyoxyethylene (35) castor oil was dissolved in 8.1ml water, and then the 8.1ml water was slowly added to the tea oil with rapid stirring to give a 3% emulsion (15% oil).

Experimental drug 19:0.3 g menthol was dissolved in 1.5g tea oil, 0.06g poloxamer 188 was dissolved in 8.14ml water, and then the 8.14ml water was slowly added to the tea oil with rapid stirring to give a 3% emulsion (15% oil).

Experimental drug 20:0.4 g menthol was dissolved in 2.5g tea oil, 0.06g poloxamer 188 was dissolved in 7.04ml water, and then 7.04ml water was slowly added to tea oil with rapid stirring to give a 4% emulsion (25% oil).

Experimental drug 21:0.7 g menthol was dissolved in 2.5g tea oil, 0.06g poloxamer 188 was dissolved in 6.74ml water, and then 6.74ml water was slowly added to tea oil with rapid stirring to give a 7% emulsion (25% oil).

Experimental drug 22:0.3 g menthol was dissolved in 1.5g tea oil, 0.2g Tween 80 was dissolved in 8ml water, and then the 8ml water was slowly added to the tea oil with rapid stirring to give a 3% emulsion (15% oil).

Experimental drug 23:0.3 g menthol was dissolved in 2.5g tea oil, 0.06g poloxamer 188 was dissolved in 7.14ml water, and then 7.14ml water was slowly added to the tea oil with rapid stirring to give a 3% emulsion (25% oil).

Experiment drug 24 1.5g lanolin was melted under 45 ℃ heating, then 0.3 g menthol was dissolved in lanolin, 0.2g tween 80 was dissolved in 8ml water and heated to 45 ℃, then the 8ml water was slowly added to lanolin with rapid stirring to give a 3% emulsion (15% oil).

Comparative example

(1) The following oil-soluble control compositions were prepared:

The oil-soluble control medicine 1:0.1 g menthol is dissolved in 9.9g olive oil, thus obtaining 1% oily liquid preparation.

The oil-soluble control medicine 2:0.2 g menthol is dissolved in 9.8g olive oil, thus obtaining the 2% oily liquid preparation.

The oil-soluble control medicine 3:0.3 g menthol is dissolved in 9.7g olive oil, thus obtaining the 3% oily liquid preparation.

The oil-soluble control medicine 4:0.4 g menthol is dissolved in 9.6g olive oil, thus obtaining 4% oily liquid preparation.

The oil-soluble control medicine is prepared by dissolving 5:0.6 g menthol in 9.4g olive oil to obtain a 6% oily liquid preparation.

The oil-soluble control medicine 6:0.8 g menthol is dissolved in 9.2g olive oil, thus obtaining the 8% oily liquid preparation.

The oil-soluble control medicine 7:1.0 g menthol is dissolved in 9.0g olive oil, thus obtaining the 10% oily liquid preparation.

The oil-soluble control medicine is prepared by dissolving 8:1.2 g menthol in 8.8g olive oil to obtain 12% oily liquid preparation.

(2) The following solid control compositions were prepared:

Solid control drug 1 according to the optimal prescription and preparation process disclosed in U.S. patent application No. US202117511238A (invention name: TREATMENT FOR DRY EYES), solid control drug 1 was prepared, which was composed of 2g of active ingredient, which was composed of ingredients of Menthoxypropanediol concentration 36.35%, menthonediol concentration 36.36%, menthol (menthol) concentration 27.27% and Caffeine concentration 0.018% in proportion, and matrix mixture (balm mixture) of beeswax 8g. The overall menthol and related mint component total concentration was about 20%.

Solid control drug 2 according to the optimal prescription and preparation process disclosed in U.S. patent application No. US202117511238A (invention name: TREATMENT FOR DRY EYES), solid control drug 2 was prepared with 6g of active ingredient consisting of 36.35% of Menthoxypropanediol, 36.36% of Menthonediol, 27.27% of Menthol (menthol) and 0.018% of Caffeine (caffeine) in proportion and 4g of beeswax as a matrix mixture (balmmixture). The overall menthol and related mint component total concentration is about 60%.

Solid control drug 3 according to the optimal prescription and preparation process disclosed in U.S. patent application No. US202117511238A (invention name: TREATMENT FOR DRY EYES), solid control drug 3 was prepared, which was composed of 0.6g of active ingredient, 36.35% in Menthoxypropanediol concentration, 36.36% in Menthonediol concentration, 27.27% in Menthol (menthol) concentration and 0.018% in Caffeine (caffeine) concentration, and 9.4g of beeswax as a matrix mixture (balmmixture). The total concentration of menthol and related mint components was 6%.

Example 2 the efficacy of the above composition in promoting tear secretion in dry eye patients was experimentally compared.

1. Purpose of experiment

The effectiveness of menthol emulsions at different concentrations in promoting tear secretion of dry eye patients was evaluated for the dose-effect relationship, and compared with the efficacy of a control drug, the skin irritation of single administration of menthol emulsions of the present invention was examined.

2. Design of experiment

Because of the menthol dermal pharmaceutical compositions of the present test, which contain menthol, olive oil and/or essence as ingredients, have been widely used in medicine and cosmetics, such as in essential balm, the concentration of menthol is 0.32g/g. Thus, it is believed that safety of at least menthol at 0.32g/g (i.e., 32%) and below this concentration range is widely accepted, as is safety for human skin.

3. Experimental method

The method comprises the steps of recruiting dry eye subjects, selecting standard (1) completely knowing the study of the test and signing informed consent, 2) age >18 years old, men and women are not limited, (3) complaints about subjective symptoms such as dry eyes, foreign body sensation, burning sensation, fatigue sensation, discomfort sensation, red eyes, vision fluctuation and the like, and (4) measuring Schirmer tear secretion the day before the test, wherein the subjects with Schirmer tear secretion less than 5mm/5min are included in the test. On the day of the test, all subjects were required to have no makeup on their faces. At the beginning of the trial, subjects were randomized into the following groups, 8/group, each:

blank drug group (i.e. blank vehicle group without menthol);

experimental drug group 1 (experimental drug 1 in example 1, emulsion containing 1% menthol);

experimental drug group 2 (experimental drug 2 in example 1, emulsion containing 1.5% menthol);

Experimental drug 3 group (experimental drug 3 in example 1, emulsion containing 2% menthol);

experimental drug group 4 (experimental drug 4 in example 1, emulsion containing 3% menthol);

experimental drug group 5 (experimental drug 5 in example 1, emulsion containing 4% menthol);

Experimental drug group 1 (experimental drug 6 in example 1, emulsion containing 5% menthol);

experimental drug group 1 (experimental drug 7 in example 1, emulsion containing 6% menthol);

experimental drug group 1 (experimental drug 8 in example 1, emulsion containing 7% menthol);

group1 oil-soluble control drugs (oil-soluble control drug 1 in example 1, containing 1% menthol);

group 2 oil-soluble control drugs (oil-soluble control drug 2 in example 1, containing 2% menthol);

group 3 oil-soluble control drugs (oil-soluble control drug 3 in example 1, containing 3% menthol);

oil-soluble control drug group 4 (oil-soluble control drug 4 in example 1, containing 4% menthol);

Group 5 oil-soluble control drugs (oil-soluble control drug 5 in example 1, 6% menthol);

Group 6 of oil-soluble control drugs (oil-soluble control drug 6 in example 1, containing 8% menthol);

group 7 oil-soluble control drugs (oil-soluble control drug 7 in example 1, 10% menthol);

group 8 oil-soluble control drugs (oil-soluble control drug 8 in example 1, containing 12% menthol);

Solid-state control drug 1 group (solid-state control drug 1 in example 1, containing 20% of active ingredient such as menthol);

Solid-state control drug group 2 (solid-state control drug group 2 in example 1, containing 60% active ingredient such as menthol);

Solid control drug 3 group (solid control drug 3 in example 1, containing 6% of active ingredient such as menthol).

After grouping, each subject was given the skin 2cm-3cm from the lower eyelid in a horizontal direction to the eyes, and the total mass of the composition of the present invention was applied at about 5 mg/under-eye skin, with an application area under each eye of about 2 square cm. After the application, each of the test subjects recorded the eye sensation and the sensation of the application site. Subjects in each group were randomized into two groups, 4/group, and Schirmer tear secretion was determined 5min and 30min after application, respectively. At the same time, subjects were scored for their compliance 30min after dosing.

Schirmer tear secretion value is an important indicator of eye comfort after treatment of dry eye patients. It is generally considered clinically that the Schirmer tear secretion value of 5min after the administration treatment can reach the normal value (namely, the Schirmer tear secretion value is more than 15 mm), and the Schirmer tear secretion value of 30min after the administration treatment can be more than 10mm, so that the patient has better comfort after the eye treatment.

The scoring criteria are 1 score (no irritation), 2 scores (slightly irritating but acceptable), 3 scores (irritating), 4 scores (strong irritation, burning but no pain), 5 scores (strong irritation, painful), an average score of 1-2 scores (including 2 scores) with good compliance, a lower score is better in compliance, an average score of 2-5 scores (excluding 2 scores) with poor compliance, and a higher score is worse in compliance.

4. Experimental results

The Schirmer tear secretion after the administration for a certain period of time was measured according to the above-described experimental method, and the measurement results are shown in table 1 below.

Table 1 test group tear secretion values (< 0.05 compared to P prior to the test)

From the specific values of tear secretion in each test group shown in Table 1, the pharmaceutical composition of the present invention exhibited a concentration dependence in the concentration range of 1% -7% in terms of the efficacy of promoting tear secretion. An emulsion with a menthol concentration of 1% has a lacrimatory effect, but does not meet the requirements of Schirmer lacrimal secretion values for clinical dry eye comfort. The emulsion of 1.5% menthol has obvious function of promoting tear secretion, and can meet the requirement of Schirmer tear secretion value for clinical treatment of dry eye comfort. With increasing concentration, the lacrimation effect is enhanced, and the maximum value of 30mm measured by Schirmer tear test paper is reached by using the emulsion with the concentration of 4% after 5 min. At a concentration of 7%, the lacrimatory secretion effect reached a maximum value of 30mm as measured by Schirmer tear test paper, and no irritation was exhibited. Referring to fig. 1, a skin irritation photograph (skin irritation was not found) of the experimental drug of the experiment of example 2, fig. 2, a skin irritation photograph (skin irritation was not found) of the experimental drug of the experiment of example 2, fig. 3, a skin irritation photograph (skin irritation was not found) of the experimental drug of the experiment of example 2, and no skin irritation was found.

From the specific values of tear secretion shown in Table 1, the oil-soluble control pharmaceutical composition exhibited concentration dependence in the concentration range of 1% -12% in terms of efficacy for promoting tear secretion. The oil-soluble control pharmaceutical compositions of 1% and 2% menthol have only weak lacrimal secretion promoting effect and do not meet the requirements of Schirmer lacrimal secretion values for clinical treatment of dry eye comfort. The oil-soluble control pharmaceutical composition containing 3% menthol has remarkable function of promoting tear secretion, and can meet the requirement of clinical treatment of the Schirmer tear secretion value of dry eye comfort. And along with the increase of the concentration, the lacrimation promoting effect is enhanced, and the maximum value of 30mm measured by Schirmer tear test paper is reached by using the oil-soluble control pharmaceutical composition with the concentration of 8% menthol after 5 min. However, when the concentration reached 12%, the lacrimation effect reached a maximum value of 30mm as measured by Schirmer tear test paper, but showed weak irritation. Thus, the tear promoting effect of the oil-soluble control pharmaceutical composition containing menthol at the same concentration was weaker than that of the emulsion of the present invention. This is probably because the emulsion composition type preparation of the present invention contains a hydrophilic phase and a lipophilic phase, is more similar to the hydrophilicity and lipophilicity of skin, is thinner and more comfortable after being applied, is less likely to block pores, is more favorable for percutaneous absorption, is favorable for acting on facial nerves to promote tear secretion, and has stronger efficacy.

Referring to fig. 4, the skin irritation photograph of the oil-soluble control drug group 3 (3% menthol) is that the skin of the application site is not reddish, and fig. 5 is that of the oil-soluble control drug group 7 (10% menthol) is that the skin of the application site is slightly reddish. Fig. 6 is a photograph of skin irritation of group 8 oil-soluble control drugs (12% menthol) showing a pronounced redness of the skin at the application site. Fig. 7 is a photograph of skin irritation of the group 1 solid control drug (containing 20% of an active ingredient such as menthol) and fig. 8 is a photograph of skin irritation of the group 2 solid control drug (containing 60% of an active ingredient such as menthol), and it can be seen that the high concentration menthol stimulates a large area of the skin application site to redly and swell. This indicates that menthol at a concentration higher than 10% is irritating to the skin and that the irritation becomes strong with increasing concentration. The oily nature, the high concentration that oil solubility brought to skin are amazing, and the human nasal wing itself is liable to produce reasons such as grease secretion, easily produces local skin inflammation and stifling problem, considers dry eye to be a chronic disease again, needs long-time medicine treatment, and weak irritation also can show the compliance that reduces patient's treatment, therefore oil solubility type control medicine 7 group and 8 group can not be as the preference of dry eye treatment.

The group 1 of solid control drugs (containing 20% of active ingredient such as menthol) had a weak lacrimatory secretion-promoting effect, probably because the group 1 of solid control drugs was a matrix of beeswax and was prepared as solid beeswax (in contrast to this, the experimental drug 11 of the present invention was an emulsion using beeswax as an oil phase matrix), and absorption of active drug ingredient such as menthol by the skin was blocked at the skin surface, and the lacrimatory secretion-promoting effect was weaker although the concentration was higher than in each experimental group. The active ingredients such as menthol have volatility, and the active ingredients such as menthol in the wax smeared on the periphery of eyes volatilize, so that the serious discomfort such as the eyes of a patient are not opened and the like is easily fumigated, and the compliance of the patient is poor. The group 2 solid control medicine (containing 60% of active ingredients such as menthol) has strong function of promoting tear secretion, but has extremely obvious irritation. The group 3 solid control drug (containing 6% of active ingredient such as menthol) is not irritating, but has very weak tear secretion promoting effect, and cannot meet the treatment expectations.

In addition, subjects in each group were studied according to the subject feeling in table 2, and subject feeling results were summarized by observing skin and understanding the medication feeling, and specific experimental results are shown in table 2.

TABLE 2 subject specific experience results (8 subjects/group)

As can be seen from Table 2, the menthol emulsion provided by the present invention is effective in promoting tear secretion of a subject in a concentration range of 1.5% -7%, and has no skin irritation. At a menthol concentration of 1%, the lacrimal secretion of the subject was promoted slightly less, and even further, the concentration was increased to 7%, and the skin irritation was not found by the experimenter. Compared with the emulsion, the oil-soluble composition and the solid waxy preparation form, the menthol emulsion has obvious drug effect, has the best effect on the activity of menthol, can completely meet the good treatment experience of dry eye patients when the concentration reaches 7 percent, has lower menthol concentration, uses less oil solvent in the emulsion and has high water phase content, can effectively reduce the greasy skin feel and the acne phenomenon after continuous and repeated use, and improves the use comfort of patients.

In conclusion, the menthol-skin oil-soluble control medicine can effectively promote tear secretion of a subject within the concentration range of 3% -10%, has no skin irritation, and has no obvious effect of promoting tear secretion of the subject when the menthol concentration is lower than 3%, and has slight skin irritation when the menthol concentration reaches 12% in part of experimenters. Whereas the solid control drug group 1 had weaker lacrimal secretion promoting effect and weak skin irritation. The solid control drug group 2 had strong skin irritation. The solid control drug 3 (menthol 6%) showed no skin irritation, but had extremely weak effect of promoting tear secretion. This is probably because the solid-state control drugs 1 to 3 are solid-state beeswax, and they are less active because they block the absorption of active drug ingredients such as menthol into the skin surface, and the active ingredients such as menthol, which are applied to the wax around the eyes, volatilize, and smoke to cause extreme discomfort such as the eyes of the patient being opened and not being opened, and the patient compliance is poor.

5. Conclusion of the experiment

The menthol skin pharmaceutical composition of the invention can effectively promote tear secretion of a subject in a concentration range of 1.5% -7%, and has no skin irritation. The medicinal composition for menthol skin has better medicinal effect than the oil-soluble control medicament with the same concentration.

Example 3 efficacy test of menthol emulsion formulated with different excipients for promoting lacrimal secretion.

1. Purpose of experiment

The effectiveness of menthol emulsions formulated with different adjuvants in promoting tear secretion in dry eye patients and skin irritation were evaluated.

2. Experimental method

The method comprises the steps of recruiting dry eye subjects, selecting standard (1) completely knowing the study of the test and signing informed consent, 2) age >18 years old, men and women are not limited, (3) complaints about subjective symptoms such as dry eyes, foreign body sensation, burning sensation, fatigue sensation, discomfort sensation, red eyes, vision fluctuation and the like, and (4) measuring Schirmer tear secretion the day before the test, wherein the subjects with Schirmer tear secretion less than 5mm/5min are included in the test. On the day of the test, all subjects were required to have no makeup on their faces. At the beginning of the trial, subjects were randomized into the following groups, 8/group, i.e.:

Experiment drug group 4 (experiment drug 4 in example 1, 3% menthol drug group formulated with olive oil as oil phase),

Experimental drug 10 group (experimental drug 10 in example 1, i.e., 3% menthol drug group formulated with castor oil as oil phase),

Experiment drug 12 group (experiment drug 12 in example 1, namely 3% menthol drug group formulated with medium chain triglyceride as oil phase),

Experiment drug 14 group (experiment drug 14 in example 1, namely 6% menthol drug group formulated with polyoxyethylene (40) castor oil as emulsifier),

Experiment drug 16 group (experiment drug 16 in example 1, namely 6% menthol drug group formulated with polyoxyethylene (35) castor oil as emulsifier),

Experimental drug 17 group (experimental drug 17 in example 1, namely, 6% menthol drug group formulated with poloxamer 188 as an emulsifier).

3. Experimental results

According to the experimental method, the Schirmer tear secretion after a certain time of administration is measured, and the detection result shows that the menthol skin pharmaceutical compositions prepared from different selected auxiliary materials have obvious effect of promoting tear secretion, and the measured tear secretion values of the menthol skin pharmaceutical compositions prepared from different auxiliary materials are not obviously different from each other after the menthol skin pharmaceutical compositions are used for 5min and 30min, and the compliance of the menthol skin pharmaceutical compositions is not obviously different from each other. The results are detailed in Table 3.

Table 3 tear secretion values of test group

4. Conclusion of the experiment

The menthol skin medicinal composition prepared from different selected auxiliary materials has obvious function of promoting tear secretion, and the functions and the stimulations of promoting tear secretion of each group have no obvious difference under proper concentration.

Example 4 test of the lacrimation secretion of menthol emulsions at different application sites.

1. The aim of the experiment is to compare the difference of the curative effects of applying menthol emulsion on the skin of different parts

2. Design of experiment

Dry eye subjects were recruited, schirmer tear secretion was measured the day prior to the test, and subjects with Schirmer tear secretion less than 5mm were enrolled in the test. At the beginning of the trial, 8 subjects were randomized into the following groups:

blank drug group (i.e. blank vehicle group without menthol);

experimental drug 4 group (experimental drug 4 in example 1) was smeared on the upper eyelid;

experimental drug 4 group (experimental drug 4 in example 1) was smeared on the lower eyelid;

Experimental drug 4 groups (experimental drug 4 in example 1) were applied to the nasal alar sites;

Experimental drug 6 group (experimental drug 6 in example 1) was smeared on the upper eyelid;

Experimental drug 6 group (experimental drug 6 in example 1) was smeared on the lower eyelid;

Experimental drug 6 group (experimental drug 6 in example 1) was applied to the nasal alar part;

The blank medicine group is simultaneously smeared on the upper eyelid, the lower eyelid and the nasal wing parts.

After grouping, the corresponding parts of each subject are respectively coated with the corresponding group of medicines, and 5 mg/position/measurement is coated. After the application, each of the test subjects recorded the eye sensation and the sensation of the application site. Subjects in each group were randomized into two groups, 4/group, and Schirmer tear secretion was determined 5min and 30min after application, respectively.

3. Experimental results:

the test results show that the effect of the medicine with the same concentration after being smeared on different parts is that the lower eyelid is stronger than the upper eyelid, and the upper eyelid is stronger than the nasal alar part, but the lacrimal secretion of a subject can be obviously promoted even if the medicine with three concentrations is administrated on the nasal alar part. The specific tear secretion values for each test group are shown in Table 4.

Table 4 test group tear secretion values (< 0.05 compared to P prior to the test)

4. Conclusion of experiment:

the effect of the medicine with the same concentration after being smeared on different parts is that the lower eyelid is stronger than the upper eyelid, and the upper eyelid is stronger than the nasal alar part, but even though the nasal alar part is provided with 5 percent of menthol emulsion (experimental medicine 6 groups), the lacrimal secretion of a subject can be obviously promoted.

Example 5 test of menthol emulsion for promoting lacrimal secretion in closed eye condition.

1. Purpose of experiment

Dry eye patients voluntarily participated in the present test, and the test under the eye-closed state was conducted to determine whether the menthol emulsion of the present invention exerts efficacy mainly through skin absorption or is mainly due to irritant tearing caused by volatilization of menthol, non-opening of volatilized menthol fumigated eyes, and the like.

2. Design of experiment

Dry eye subjects were recruited, schirmer tear secretion was measured the day prior to the test, and subjects with Schirmer tear secretion less than 5mm were enrolled in the test. At the beginning of the trial, subjects were randomized into the following groups, 4/group, respectively:

blank drug group (i.e., drug group without menthol);

experimental drug 3 group (experimental drug 3 in example 1);

Experimental drug 7 group (experimental drug 7 in example 1).

The self-adhesive transparent laser postoperative protection light-transmitting single eye mask is used for blocking all the ventilation holes on the eye mask by using a medical adhesive tape. After grouping, both eyes of each subject were covered with an eye mask, and then when the subject was about to be coated with the drug, the eyes were gently closed until tear secretion was measured, and the eyes were not opened. After the eyes of the subjects are closed, medicines are coated, and when medicines are coated, medicines of corresponding groups are coated on the lower eyelid of each subject, and the coating is about 5 mg/position/measurement. The repeated delivery of the subjects gently closes eyes for rest and does not open eyes. After the application, each of the test subjects recorded the eye sensation and the sensation of the application site after the end of the experiment. Schirmer tear secretion was measured 5min after application of each group of subjects.

3. Experimental results

Experimental results show that under the condition of eye shielding and eye closure of a subject, the menthol emulsion provided by the invention can obviously promote tear secretion of the subject, and the quantity of the tear secretion is not obviously different from that recorded in the example 2. Menthol has certain volatility, and the test result proves that the medicine of the invention does not stimulate the ocular surface through the volatilized menthol to promote tear secretion. The specific tear secretion values for each test group are shown in Table 5.

Table 5 test group tear secretion values (< 0.05 compared to P prior to the test)

4. Conclusion of the experiment

The medicine of the invention does not stimulate ocular surface through volatilized menthol to promote tear secretion, and is supposed to play a role in drug effect mainly through skin absorption.

EXAMPLE 6 evaluation of greasiness of menthol dermal pharmaceutical compositions and control drugs

1. Purpose of experiment

Evaluation of skin greasiness of menthol emulsion after continuous multiple use

2. Design of experiment

Greasiness refers to the excessive greasy feel of a dermatological product in use, known as "oily" which affects the compliance of the product with use. Dry eye subjects were recruited, and the day before the test required no cosmetic such as emollient, and all subjects were cleaned 1h before the test day, and the test required no rubbing of the smears such as wet tissues during the test. The 24 subjects (lower eyelid application site at 48) were enrolled in the trial, with random lower eyelid application of test drug 3 (2% menthol, 8% olive oil), test drug 20 (4% menthol, 25% olive oil), and oil-soluble control drug 4 (4% menthol, olive oil dissolved) applied to the lower eyelid. The above is applied once every two hours, and 6 times are continuously applied (dry eye patients are usually treated with drugs as needed, and the use frequency is 6 times or more than 6 times a day) to simulate the use of the emulsion of the invention in one day. 2h after the 6 th application, observing the greasy skin condition of the lower eyelid of the subject, and grading the comfort degree of the lower eyelid of the two eyes of the subject, wherein 1 is the most refreshing, no greasy feeling is caused, 9 is the most greasy feeling, and the subject is free to select which medicament is prone to be used subsequently.

3. Experimental results

The skin of the subject on which the oil-soluble control drug 4 was smeared was visually oily to the skin of the lower eyelid smears, whereas the skin of the subject on which the lower eyelid smears were smeared was only faintly oily. Subjects had a heavy greasy feel on the skin from the group 4 of the oil-soluble control drug, especially on the 5 th and 6 th times of application, had a strong greasy feel on the skin, had an impulse to wash the face or wipe with a wet wipe or rub with the hands, and had a greasy feel score of 8.5±0.5 (see fig. 9).

The skin of the lower eyelid smeared part of the experimental drug 20 (containing 25% of tea oil) has a slightly greasy feel, and the overall refreshing degree is far better than that of the oil-soluble control drug 4 group (containing 96% of olive oil), and the greasy feel score of the subjects aiming at the experimental drug 20 is 2.9+/-0.5 (P <0.05 compared with the oil-soluble control drug 4 group). The skin of the lower eyelid smeared part of the experimental drug 3 (containing 8% of olive oil) has no greasy feel, the overall refreshing degree is far better than that of the oil-soluble control drug 4 group, and the greasy feel score of the subjects aiming at the experimental drug 3 is 1.8+/-0.8 (P <0.05 compared with the oil-soluble control drug 4 group and P <0.05 compared with the experimental drug 20 group). All subjects selected the order of use of the preferred experimental drug 3, the next experimental drug 20, and the 3 rd selection was the oil-soluble control drug 4.

4. Conclusion of the experiment

The menthol emulsion provided by the invention has the advantages that the greasy feeling of a subject after the menthol emulsion is continuously used for many times is weaker than that of an oil-soluble control medicament, and the comfort feeling and the refreshing feeling are better than those of the oil-soluble control medicament.

EXAMPLE 7 evaluation of drug efficacy of menthol emulsion and oil-soluble control drug after multiple administration

1. Purpose of experiment

Evaluating whether the efficacy of menthol emulsion after continuous multiple use has a decreasing trend

2. Design of experiment

Dry eye is a chronic disease that is administered for a long period of time and requires multiple consecutive administrations per day. The oil-soluble liquid preparation is used for a patient for a plurality of times continuously, the skin feel of the patient is poor, the patient has the problem of greasy skin after the patient uses the oil-soluble liquid preparation for a plurality of times continuously, and after the oil-soluble liquid preparation is used for a plurality of times continuously, the medicine application part is covered by an oil layer, the oil molecules block pores, and the curative effect is reduced after the oil-soluble liquid preparation is used for a plurality of times continuously. Dry eye subjects were recruited, and the day before the test required no cosmetic such as emollient, and all subjects were cleaned 1h before the test day, and the test required no rubbing of the smears such as wet tissues during the test. Schirmer tear secretion was measured prior to dosing in the subjects. Subjects were then divided into 4 groups of 8 each, with experimental drug 3 (2% menthol, 8% olive oil) applied to the lower eyelid of group 1, experimental drug 23 (3% menthol, 25% tea oil) applied to the lower eyelid of group 2, oil-soluble control drug 3 (3% menthol, 97% olive oil dissolved) applied to the lower eyelid of group 3, oil-soluble control drug 3 (3% menthol, 97% olive oil dissolved) applied to the lower eyelid of group 4. Groups 1,2 and 3 above were each applied once every two hours, and 6 times in succession (dry eye patients were usually treated with the drug as needed, with a frequency of use of 6 times or more per day) to simulate the use of the drug of the present invention per day. The group 4 is applied every two hours, and is continuously applied for 6 times by lightly wiping and degreasing with a mind act skin cleaning and degreasing wet tissue (special for cleaning) 30min before the second application. Each of the above groups was divided into two groups, and Schirmer tear secretion was measured 1 time after the application and 5 minutes after 6 times of the application, respectively.

3. Experimental results

Subjects in group 3 (oil-soluble control drug group 3, 3% menthol, 97% olive oil dissolved) had lower Schirmer values after the 6 th application than after the first administration (P < 0.05). And subjects in group 4 (oil-soluble control drug 3, 3% menthol, 97% olive oil dissolved, external wet wipe clean) had a Schirmer value after 6-application dose comparable to the Schirmer value after the first dose (P > 0.05).

Subjects in group 2 (experimental 23,3% menthol, 25% tea oil) had slightly lower Schirmer values after the 6 th application than after the first application but no statistical differences (P > 0.05). Whereas subjects in group 1 (experimental 3,3% menthol, 8% olive oil) had a Schirmer value after the 6 th application of the dose comparable to the Schirmer value after the first dose (P > 0.05), the specific results are detailed in table 6.

Table 6 lacrimal secretion values of test group (< 0.05 compared to 5min after the first application)

4. Conclusion of the experiment

As can be seen from the 3 rd combination and the 4 th group experiments, the continuous repeated use curative effect of the oil-soluble liquid preparation can be reduced, if the efficacy is to be maintained, the oil layer which is remained after repeated use is cleaned by wet tissues and the like, and the menthol emulsion prepared by the invention also has the obvious problem of the continuous repeated use curative effect reduction.

Example 8 evaluation of safety of menthol emulsion and control drug for long-term skin administration.

1. Purpose of experiment

The safety of the menthol emulsion of the invention for long term administration (28 days) was evaluated using a mouse animal experiment.

2. Design of experiment

Female C57BL/6 mice were randomly divided into groups of 6 mice each, i.e.:

healthy murine controls (vehicle blank administration);

Experimental drug 7 group (experimental drug 7 in example 1, menthol 6%, olive oil 25%);

experimental drug group 8 (experimental drug 8 in example 1, menthol 7%, olive oil 25%);

group 7 of oil-soluble control drugs (oil-soluble control drug 7, menthol 10% in example 1);

Group 8 of oil-soluble control drugs (oil-soluble control drug 8, menthol 12% in example 1);

Solid state control drug group1 (solid state control drug 1, menthol 20% in example 1);

solid control drug group 2 (solid control drug 2, menthol 60% in example 1).

The corresponding drugs were applied to the skin of about 4mm on the lower eyelid of the mice, about 5mg each time on the lower eyelid of each eye, and the healthy control group was given a blank vehicle 6 times daily (8 points, 11 points, 13 points, 15 points, 17 points, 20 points), about 5mg each time on the lower eyelid of each eye. 30min after the last dose on day 28, the skin irritation at the site of application of each group of mice was examined and observed and scored according to the skin irritation response scoring criteria (scoring criteria are shown in Table 7).

3. Experimental results

TABLE 7 skin irritation response scoring criteria

Healthy control mice are smeared under the eyelid by a blank solvent, and after 28 days of continuous smearing, the skin is not stimulated by abnormal red swelling and the like. In addition, the oil-soluble control drug group 7 (10% menthol drug group) mice were also not abnormal throughout the observation.

After each administration, the mice in the oil-soluble control drug group 8 (12% menthol drug group) were occasionally left with the front paws of the mice to grasp the drug-coated parts, but only showed a transient reaction, no continuous scratching of the mice, continuous administration for 28 days, no other abnormalities were seen except the red coated parts.

Mice in group 1 of solid control drug (solid control drug 1 in example 1, i.e., menthol and related mint component concentration of about 20%) were seen to have had their forepaws removed from the drug application site after each administration, and occasionally had their continuous scratching, continued administration for 28 days, and the application site was slightly reddish, with no other abnormalities.

Immediately after the administration of the mice in the group 2 of solid control drugs (the group 2 of solid control drugs in example 1, namely, the group containing 60% of active ingredients such as menthol) the front paws of the mice were found to be in a position to catch the drug severely, and the coated parts were red and swollen after the administration of the mice for a plurality of times, which reflects the strong irritation of menthol at the concentration to the lower eyelid of the mice. The results of the specific group of stimuli are summarized in Table 8.

Table 8 evaluation of long-term administration irritation in mice

4. Conclusion of the experiment

The 7% menthol emulsion of the invention has no skin irritation even if used for a long time. 10% of the oil-soluble control drugs had no long-term skin irritation, 12% of the oil-soluble control drugs had mild skin irritation after long-term use, and the solid-state control drug 1 group (solid-state control drug 1 in example 1) had mild skin irritation after long-term use, and the solid-state control drug 2 group (solid-state control drug 2 in example 1) had strong skin irritation after long-term use.

Example 9 experiments of menthol emulsion to promote tear secretion in dry eye model mice.

1. Purpose of experiment

The effectiveness of the menthol emulsion of the present invention in promoting tear secretion and treating dry eye was evaluated by using a mouse animal experiment.

2. Design of experiment

(1) Grouping modeling and processing of experimental animals

The dry stress environment box was set at constant temperature (24.+ -. 2 ℃), constant humidity (30.+ -. 5%), constant wind speed (2.5.+ -. 0.5 km/s). Female C57BL/6 mice were randomly divided into groups of 6 mice each, i.e.:

Healthy murine controls (normal saline for modeling, blank vehicle oil for treatment);

Blank (blank drug group without menthol after modeling);

experimental drug group 2 (2,1.5% menthol after modeling with experimental drug in example 1);

experimental drug 3 group (experimental drug 3,2% menthol in example 1 after modeling);

experimental drug group 5 (experimental drug 5,4% menthol in example 1 after modeling);

Experimental drug 7 group (7, 6% menthol with experimental drug in example 1 after modeling);

experimental drug group 8 (experimental drug 8,7% menthol in example 1 after modeling);

oil-soluble control drug 3 group (3, 3% menthol with oil-soluble control drug after modeling);

Oil-soluble control drug group 4 (oil-soluble control drug 4,4% menthol after modeling);

Oil-soluble control drug group 5 (5, 6% menthol with oil-soluble control drug after modeling);

Group 6 of oil-soluble control drugs (6, 8% menthol with oil-soluble control drug after modeling);

oil-soluble control drug group 7 (modeling followed with oil-soluble control drug 7,10% menthol);

oil-soluble control drug group 8 (oil-soluble control drug 8,12% menthol after modeling);

solid control drug group 1 (solid control drug 1,20% menthol after modeling);

solid control drug group 2 (solid control drug 2,60% menthol after modeling).

Mice in the healthy mouse control group were subcutaneously injected with 0.2mL of physiological saline into the hind limbs 4 times daily (9, 12, 15, 18) and kept in a normal environment with 55% ± 10% relative humidity for 5 days, for 5 consecutive days. Mice in groups other than the healthy mouse control group were injected subcutaneously into the hind limbs 6 times daily (8, 11, 13, 15, 17, 20) with 2.5mg/mL scopolamine, 0.2mL each time, and placed in a dry stress environment for 5 consecutive days. At the same time of modeling, the corresponding drugs of each group were applied to the skin of about 4mm at the lower eyelid of the mouse, about 5mg each time at the lower eyelid of each eye, and the healthy control group was given a blank vehicle, 6 times per day, about 5mg each time at the lower eyelid of each eye. Each index of each group of mice was examined 30min after the last dose on day 5.

(2) Tear secretion measurement

Phenol red cotton thread is a common index for detecting tear secretion, and the tear secretion condition of a mouse can be quantified by the length of the phenol red cotton thread changing to red. Modeling tear secretion was measured for each group of mice on the fifth day. The lower eyelid of the mouse is gently pulled up by using an ophthalmic forceps at the same time under the condition of gas anesthesia, a phenol red cotton thread is inserted into the inner side of a conjunctival fornix, the lower eyelid is taken out after being placed for 20 seconds, and the length of the reddened cotton thread is measured by using a ruler and recorded.

(3) Fluorescein sodium staining

Corneal epithelial barrier function injury was observed by corneal fluorescein staining. Mice were anesthetized with gas and then 2.5 μl of 0.25% sodium fluorescein dye was topically applied to the cornea. Manual flash 5 times, 1mL saline flush with 1mL syringe, and a rough photograph under cobalt blue light of a slit microscope. For grading of fluorescein staining, the cornea was divided into four quadrants, each scored. The 4 scores were added to give the final score for each eye (minimum 0 and maximum 16). The fluorescein scores in each quadrant were none, 0, less than 30 spots with light punctate staining, 1, more than 30 spots with dense punctate staining, 2, severe diffuse staining with no positive plaques, 3, fluorescein positive plaques, 4.

3. Experimental results

After 5 days of modeling, sodium fluorescein was dropped onto the mouse ocular surface, and then photographed under a slit lamp with cobalt blue light to score the photographs clinically. The results showed an increase in corneal fluorescein score (12.37.+ -. 2.64VS 0.57.+ -. 0.21) for the placebo group, suggesting a significant defect in the corneal epithelium. However, in contrast, the corneal fluorescein staining score was significantly decreased after 5 days of treatment (fig. 10 and 11) and tear secretion was also significantly increased (fig. 12) for each experimental drug-treated group. Healthy control mice had no difference in corneal epithelial scores after application of blank vehicle under the eyelid as compared to before the start of the experiment. And after the five groups of menthol emulsion provided by the invention are administrated, the mice have no manifestation reflecting the irritation of the medicine, such as the positions where the medicine is coated by the front paws.

After mice were dosed with the oil-soluble control drug group 8 (oil-soluble control drug group 8 in example 1, i.e., 12% oil-soluble menthol drug group), the mice were presented with the forepaw of the mice at the site of drug application, reflecting the slight irritation of menthol at this concentration to the lower eyelid of the mice. After the mice in the group 1 (group containing 20% of active ingredients such as menthol) are dosed with the solid control drug, the front paws of the mice can be seen to remove the drug coating part, and the irritation of the menthol with the concentration to the lower eyelid of the mice is reflected. Immediately after the administration of the mice in the group 2 (group containing 60% of active ingredients such as menthol) of the solid control drug, the front paws of the mice were found to be at the place where the drug was applied, reflecting the strong irritation of menthol at this concentration to the lower eyelid of the mice.

4. Conclusion of the experiment

From the above experimental results, the menthol emulsion provided by the invention has obvious curative effect of promoting tear secretion on a xerophthalmia model mouse within the menthol concentration range of 1.5% -7%, does not cause uncomfortable reactions such as irritation and the like of the mouse, and has good use safety.

Example 10 study of mechanism of menthol emulsion to promote tear secretion.

1. Purpose of experiment

The mechanism of the menthol emulsion for promoting tear secretion is disclosed by using a mouse animal experiment.

2. Design of experiment

Female C57BL/6 mice were randomly divided into 8 mice per group, i.e.:

healthy murine control group;

healthy murine treatment group (experimental drug 5 in example 1, 4% menthol drug group);

blank treatment group (i.e., blank vehicle group without menthol);

experimental drug group 2 (experimental drug group 2 in example 1, i.e., 1.5% menthol drug group);

Experimental drug 5 group (experimental drug 5 in example 1, i.e. 4% menthol drug group);

experimental drug group 8 (experimental drug 8 in example 1, i.e. 7% menthol drug group).

Nerve blocking mouse modeling principle that compound lidocaine and carbamazepine are nerve blocking agents, and after combined administration, the nerve in the periocular skin is blocked. With a delayed nerve block, tear secretion is reduced compared to normal mice.

Healthy murine control mice and healthy murine treatment mice were intraperitoneally injected 1 time per day (9 am) with 0.2mL of physiological saline for 5 consecutive days. The other groups of mice were intraperitoneally injected with carbamazepine at a dose of 30mg/kg 1 time per day (9 am) and coated with 5% of the compound lidocaine cream (national drug standard 2063466, manufactured by the same pharmaceutical company, inc.) at a position around the eyes 2 times per day (9 am, 18 am), 250 mg/time, and the coated part of the compound lidocaine was composed of the part coated with the menthol skin drug composition, and the coated area was at least twice the area coated with the menthol skin drug composition, and was continued for 5 days per day. On the fourth day of the experiment, all mice measured tear secretion.

On the fifth day of the experiment, 1h after the compound lidocaine is coated for the second time, the part coated with the compound lidocaine is gently rubbed by a cotton swab wetted by normal saline, each group of corresponding experiment drugs are respectively given 15min after the rubbing is finished, each group of mice is randomly divided into two groups of 4 mice, 5min and 30min after the administration, one group of mice is randomly selected respectively, and tear secretion of the mice is measured by using a phenol red cotton thread. On the fifth day of the experiment, the healthy rat control group mice were coated with a blank solvent solution, the healthy rat treatment group mice were coated with an experiment drug 5, the other groups of mice were dosed according to the experimental group, then each group of mice was randomly divided into two groups of 4 mice, 5min and 30min after dosing, one group was randomly selected, and the tear secretion of the mice was measured by phenol red cotton line.

3. Experimental results

As shown in fig. 13, the tear secretion of healthy control mice was not significantly changed after the healthy control mice were coated under the eyelid with the blank vehicle, while the tear secretion of healthy mice was significantly increased after the treatment with drug 5 (4% menthol emulsion). The compound lidocaine and carbamazepine are nerve blocking agents, nerve in the skin around eyes is blocked, tear secretion is reduced compared with that of a normal mouse, and even if menthol emulsion of the invention with different concentrations is further administered, the tear secretion of the mouse is not obviously increased. This demonstrates that menthol compositions of the present invention promote tear secretion primarily by acting on the facial nerves of animals.

4. Conclusion of the experiment

The medicine of the present invention promotes tear secretion mainly by acting on facial nerves.

Example 11 menthol emulsion promotes self tear secretion compared to commonly used drugs on the market.

1. Purpose of experiment

The difference of the curative effect of the menthol emulsion of the invention and the common medicines for treating xerophthalmia on the market is compared through a mouse experiment.

2. Design of experiment

3 Drugs commonly used for dry eye treatment, which have been marketed, are 0.3% sodium hyaluronate eye drops (trade name: alic), 0.4% polyethylene glycol eye drops (trade name: plausin), 3% diquafosol sodium eye drops (trade name: liai), as control drugs.

healthy murine control group;

blank (i.e., blank vehicle without menthol);

Experimental drug group 5 (experimental drug 5 in example 1, i.e. 4% menthol emulsion);

control drug 4 group (0.3% sodium hyaluronate eye drops group);

Control drug group 5 (0.4% polyethylene glycol eye drops);

control drug 6 group (3% diquafosol sodium eye drops).

Healthy murine control mice were subcutaneously injected into the hind limbs 4 times daily (9, 12, 15, 18) with 0.2mL saline and kept in a normal environment with 55% ± 10% relative humidity for 5 days, for 5 consecutive days. The rats of the other groups except the healthy rat control group were subcutaneously injected with 2.5mg/mL scopolamine into the hind limbs 6 times daily (8, 11, 13, 15, 17, 20), 0.2mL each time, and placed in a dry stress environment for 5 consecutive days. While modeling, the applied drug was applied to the skin of about 4mm at the lower eyelid of the mouse, about 5mg each time at the lower eyelid of each eye. The healthy control group was given a blank vehicle 4 times daily, about 5mg each time applied to the lower eyelid of each eye. The control drug 4 group (0.3% sodium hyaluronate eye drops), the control drug 5 group (0.4% polyethylene glycol eye drops) and the control drug 6 group (3% quartenary sodium eye drops) were administered 4 times daily, with about 5 μl (about 5 mg) per eye drop. The tear secretion measurements and corneal fluorescein staining were measured for each group of mice 30min after the last dose on day 5, and the tear secretion measurements were measured for each group of mice 2h and 4 h.

3. Experimental results

The results showed an increase in corneal fluorescein score for the blank group, suggesting a significant defect in the corneal epithelium. In contrast, however, the cornea was not significantly defective after 5 days of treatment for each of the experimental and control drug groups, and the fluorescein staining score was significantly decreased (fig. 14 and 15). The measured tear secretion values at three time points of 30min, 2h and 4h after the administration of the experimental drug 5 group showed a decreasing trend with time, but the tear secretion was significantly increased compared with the blank treatment group, indicating that the menthol emulsion of the present invention had the effect of stimulating and promoting the recovery of the tear function of the dry eye model mice to a certain extent. The three control drugs have no obvious increase of tear secretion at three time points after administration, which indicates that the three control drugs have no obvious effect on restoring the function of self tear secretion of the xerophthalmia mice, the action mechanism is mainly to play a role of an exogenous simulated tear supplement, after eye drops, the measured simulated tear of the ocular surface (strictly speaking, the exogenous dripped simulated tear is dripped eye drops) is increased under the exogenous liquid supplement action, but the exogenous dripped simulated tear is lost on the ocular surface and the ocular surface begins to become dry and astringent again along with the extension of time. The experiment fully shows that the low-concentration menthol medicine of the invention and the existing artificial tear type eye drops belong to different treatment mechanisms for treating xerophthalmia, and the medicine has more remarkable treatment effect for acting on facial nerves to promote the organism to produce tear.

4. Conclusion of the experiment

The low-concentration menthol medicine is a different therapeutic mechanism from the existing artificial tear type eye drops for treating xerophthalmia, and the therapeutic effect of the medicine is more remarkable. The medicine provided by the invention has the curative effect of stimulating tear function of a dry eye model mouse to achieve a certain degree of recovery, and 3 medicines widely used in clinic have no effect.

It should be noted that the above embodiments are merely for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that the technical solution described in the above embodiments may be modified or some or all of the technical features may be equivalently replaced, and these modifications or substitutions do not make the essence of the corresponding technical solution deviate from the scope of the technical solution of the embodiments of the present invention.

Claims

1. Use of low-concentration menthol as the sole active ingredient in the preparation of a transdermal emulsion for facial administration for treating dry eye, wherein the transdermal emulsion contains low-concentration menthol as the sole active ingredient and an oil phase matrix, an emulsifier and an aqueous phase, wherein the mass proportion of menthol is 1.5%-7%, the mass proportion of the oil phase matrix is 3%-25%, the mass proportion of the emulsifier is 0.05%-3%, and the balance is an aqueous phase;

The oil phase matrix is one or more selected from tea oil, olive oil, soybean oil, coconut oil, sunflower oil, corn oil, avocado oil, shea butter, grape seed oil, almond oil, castor oil and jojoba oil, lanolin, beeswax, medium chain triglycerides, isopropyl palmitate, safflower oil, stearyl alcohol, stearic acid, cetyl alcohol, cholesterol, silicone oil, liquid paraffin;

The emulsifier is at least one selected from lecithin, polyoxyethylene 35 castor oil, polyoxyethylene 40 castor oil, polyoxyethylene 40 hydrogenated castor oil, Tween 80, Span 80, poloxamer 188, poloxamer 407, sodium stearate, potassium stearate, sodium lauryl sulfate, sodium hexadecyl sulfate, cetyl alcohol, polyoxyethylene 40 stearate, polyoxyethylene lauryl ether, and sucrose fatty acid ester.

2. The use according to claim 1, characterized in that the menthol is natural menthol or synthetic menthol.

3. The use according to claim 1, characterized in that the emulsion consists of 2%-6% menthol, 5%-20% oil phase base, 0.05%-3% emulsifier and the balance water phase.

4. The use according to claim 1, characterized in that the aqueous phase is water, or an aqueous solution in which a water-soluble auxiliary agent is dissolved.