CN114601959A

CN114601959A - Medical skin care dressing and preparation method and application thereof

Info

Publication number: CN114601959A
Application number: CN202111385369.5A
Authority: CN
Inventors: 张景海; 赵华军; 燕姿辰
Original assignee: Shandong Rongyuankang Medical Technology Co ltd
Current assignee: Shandong Rongyuankang Medical Technology Co ltd
Priority date: 2020-11-23
Filing date: 2021-11-22
Publication date: 2022-06-10

Abstract

The present invention provides: a medical skin care dressing comprises a nonionic surfactant with a concentration of not less than 1 wt%, sodium hyaluronate with a concentration of not less than 1 wt%, sodium carboxymethylcellulose with a concentration of not less than 0.1 wt%, collagen, alginate and chitosan with a total concentration of not less than 0.1 wt%, and water, and a preparation method and application thereof. The medical skin care dressing can dissolve and load various skin external medicines which are clinically used at present, can promote the transdermal absorption rate of the medicines and plays an auxiliary treatment role in caring the skin.

Description

Medical skin care dressing and preparation method and application thereof

Technical Field

The invention relates to the field of medical instruments, in particular to the field of medical consumables.

Background

The dressing refers to auxiliary materials used besides the main material of the article. Traditional dressings are mainly dry gauze and oil gauze. Modern wound dressings include interactive wound dressings, calcium alginate dressings, silver dressings, foam dressings, hydrocolloid dressings, hydrogel dressings, and the like.

The interactive wound dressing contains acrylate polymer with super-efficient absorption, continuously releases ringer's solution for 24 hours after being activated by ringer's solution, continuously, actively and powerfully cleans wounds, and is suitable for all open wounds (except narrow and deep wounds), including infectious wounds and healing stages, particularly the debridement stage.

The calcium alginate dressing is a mixture of alginic acid and calcium ions, forms a smooth gel after contacting with wound exudate and oozing blood, protects the wound surface, and effectively promotes wound healing.

The silver dressing is an ideal anti-infection dressing, silver ions of which can directly kill bacteria, control wound infection, accelerate wound healing and remove peculiar smell generated by bacteria.

Most of foam dressings are formed by foaming water molecule materials, and a polyurethane semipermeable membrane is covered on the surface of the foam dressing. High exudate absorption capacity, efficient exudate management capacity, high hydrophilic materials, reduced risk of wound adhesion, promoted wound healing, and foam pad buffering external pressure.

The hydrocolloid dressing adopts a closed moisturizing principle, hydrophilic CMC particles are contacted with wound exudation, a layer of moist gel is formed on the surface of a wound surface, a moist environment is continuously created, and the hydrocolloid dressing is not adhered to the wound surface. For example, patent document 1 discloses a hydrocolloid dressing. The dressing has high liquid absorption, and can be applied to dressing of high exudation wound surface. Patent document 2 discloses an amorphous hydrocolloid dressing containing a liposome. The dressing has active exudation resisting and antibacterial effects, and can promote healing.

The hydrogel dressing is a hydrophilic polyurethane polymer with high water content, automatically adjusts the wettability of a wound, has a small capacity of absorbing seepage, does not stick to the wound and is easy to remove. For example, patent document 3 discloses an alginate-based hydrogel dressing. The dressing has the advantages of good moisture retention, excellent mechanical properties and good transparency. Patent document 4 discloses an antibacterial hydrogel dressing which can wrap a drug therein by utilizing its porous structure, directly acts on a wound surface, and has a bactericidal ability itself.

However, none of the prior arts discloses a dressing which can dissolve and carry various skin external drugs and can promote the transdermal absorption of the skin external drugs and exert an auxiliary treatment effect for caring the skin.

Particularly, the incidence rate of the infantile hemangioma is increased year by year, and epidemiology indicates that the incidence rate of the infantile hemangioma is 4-10% in China and the incidence rate is high in coastal areas. 70% of good-haired parts of infantile hemangioma are in the head and face, and the beauty is directly influenced. There are many methods for treating infantile hemangioma, such as invasive treatment by laser, freezing, electric burn, chemical agent burn, oral drug therapy, and topical drug therapy. The invasive treatment has great pain and leaves traces. Oral medication, such as antineoplastic drugs, propranolol (beta receptor blocker), and the like, but the growth and development of the children are adversely affected because the tissues and organs of the children are mature. The external drug therapy includes the use of liquid, gel, ointment formulations containing beta blockers.

The application of the liquid preparation requires that the treatment liquid medicine containing the beta receptor blocker is soaked in gauze or cotton balls and is adhered to the surface of the infantile hemangioma by medical adhesive plaster. The transdermal properties of the drug are uncertain, and therefore, the dosage cannot be scientifically estimated.

Although the ointment preparation or the gel preparation is reported, the medicine for treating infantile hemangioma is prepared into the ointment preparation or the gel preparation. Because the application and approval period of the exclusive external medicine is as long as several years, the investment is high, and many medicines belong to the medicines of the young and the public, few pharmaceutical factories are willing to declare the external medicine for treating the infantile hemangioma. As a result, although there is a report on an external preparation for treating infantile hemangioma, no external preparation for treating infantile hemangioma is actually used in clinical practice, and thus patients cannot receive external treatment with less damage and high curative effect.

In addition, some external skin drugs, although used in an ointment preparation or a gel preparation, have room for further improvement in their transdermal therapeutic effects against diseases such as infantile hemangioma.

It is considered that, up to now, no medical skin care dressing has been proposed in any prior art, which can dissolve and load various skin external drugs, and improve the transdermal absorption rate of the drugs, and at the same time, has an auxiliary treatment effect.

Documents of the prior art

Patent document

Patent document 1: CN108339145B

Patent document 2: CN106474526B

Patent document 3: CN106492260B

Patent document 4: CN106693042B

Disclosure of Invention

Problems to be solved by the invention

The present invention has been made in view of the above problems, and an object of the present invention is to provide a medical skin care dressing which can dissolve and load various skin external drugs that have been used in clinical practice, and can promote the transdermal absorption rate of these drugs and exert an auxiliary therapeutic effect for caring skin.

That is, the present invention aims to: provided is a medical skin care dressing capable of dissolving and loading a known drug for external application to the skin and improving the transdermal therapeutic effect of the drug. Further objects of the invention are also: provides a medical skin care dressing which can dissolve and load an external medicine for treating infantile hemangioma and improve the transdermal treatment effect of the medicine.

Means for solving the problems

In order to achieve the above object, the present invention provides: a medical skin care dressing comprising a nonionic surfactant at a concentration of not less than 1 wt%, sodium hyaluronate at a concentration of not less than 1 wt%, sodium carboxymethylcellulose at a concentration of not less than 0.1 wt%, at least one selected from the group consisting of collagen, alginate and chitosan at a combined concentration of not less than 0.1 wt%, and water, relative to the entirety of the medical skin care dressing.

ADVANTAGEOUS EFFECTS OF INVENTION

The medical skin care dressing improves the transdermal treatment effect of the existing clinical external skin medicines (such as medicines for treating infantile hemangioma, medicines for treating infantile vascular malformation, medicines for treating chloasma, medicines for promoting local healing, or local pain-relieving medicines), and particularly makes up the defects of the existing external clinical medicines for treating infantile hemangioma and medication defects.

Drawings

FIG. 1 is a standard curve diagram of tranexamic acid standard;

FIG. 2 is an HPLC chromatogram of tranexamic acid standard;

FIGS. 3-16 are sequential in vitro transdermal absorption results for samples A01, A02, A03, A04, A05, A06, A07, B01, B02, B03, B04, B05, B06, and B07;

FIG. 17 is a standard graph of a sirolimus standard;

FIG. 18 is an HPLC chromatogram of a sirolimus standard;

FIGS. 19-32 show in sequence the in vitro transdermal absorption results for samples E01, E02, E03, E04, E05, E06, E07, F01, F02, F03, F04, F05, F06, and F07;

FIGS. 33-41 show the results of in vitro transdermal absorption of samples IA 01, IA 02, IA 03, IIB 01, IIB 02, IIB 03, IIIC 01, IIIC 02, and IIIC 03 in sequence;

FIGS. 42-44 are the film forming results for formulation A, formulation B, and formulation C.

Detailed Description

[ medical skin-Care dressing of the invention ]

The medical skin care dressing of the present invention comprises a nonionic surfactant at a concentration of not less than 1 wt%, sodium hyaluronate at a concentration of not less than 1 wt%, sodium carboxymethylcellulose at a concentration of not less than 0.1 wt%, at least one selected from the group consisting of collagen, alginate and chitosan at a total concentration of not less than 0.1 wt%, and water, with respect to the entirety of the medical skin care dressing.

The nonionic surfactant is a polyoxyethylene type nonionic surfactant, and preferably, the nonionic surfactant comprises poloxamer.

The sodium hyaluronate comprises high molecular sodium hyaluronate, medium molecular sodium hyaluronate and small molecular sodium hyaluronate,

preferably, the content of the high molecular sodium hyaluronate is 0.01-2 wt%, the content of the medium molecular sodium hyaluronate is 0.01-5 wt%, and the content of the small molecular sodium hyaluronate is 0.01-5 wt% of the whole medical skin care dressing.

The molecular weight of the high molecular sodium hyaluronate is more than or equal to 1800 kDa.

The molecular weight of the medium molecular sodium hyaluronate is 1000-1800 kDa.

The molecular weight of the small molecule sodium hyaluronate is 10-1000 kDa.

Preferably, the nonionic surfactant is present in a concentration of 1 to 10 wt%, preferably 2 to 8 wt%, more preferably 3 to 6 wt%, for example, in a concentration of 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, 5 wt%, 5.5 wt%, 6 wt%, 6.5 wt%, 7 wt%, 7.5 wt%, 8 wt%, 8.5 wt%, 9 wt%, 9.5 wt%, 10 wt%.

Preferably, the sodium hyaluronate is in a concentration of 1 to 10 wt%, preferably 1 to 5 wt%, more preferably 1 to 3 wt%, for example, in a concentration of 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, 5 wt%, 5.5 wt%, 6 wt%, 6.5 wt%, 7 wt%, 7.5 wt%, 8 wt%, 8.5 wt%, 9 wt%, 9.5 wt%, 10 wt%.

Preferably, the concentration of the sodium carboxymethylcellulose is 0.1-10 wt%, preferably 0.5-2 wt%, more preferably 0.5-1 wt%, for example, the concentration of the sodium carboxymethylcellulose is 0.1 wt%, 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, 5 wt%, 5.5 wt%, 6 wt%, 6.5 wt%, 7 wt%, 7.5 wt%, 8 wt%, 8.5 wt%, 9 wt%, 9.5 wt%, 10 wt%.

Preferably, the total concentration of at least one selected from the group consisting of collagen, alginate and chitosan is 0.1% to 10% by weight, preferably 0.4 to 4% by weight, more preferably 0.5 to 2% by weight, for example, the total concentration of at least one selected from the group consisting of collagen, alginate and chitosan is 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10% by weight.

The amount of water added is not particularly limited as long as it can be used to form the medical skin care dressing of the present invention, and for example, the amount of water added is 80 wt% or more, 90 wt% or more, 97.8 wt% or less, 96 wt% or less, or the like, relative to the entire dressing.

The invention also provides a method for preparing the composition, which is characterized by comprising the following steps: the method comprises the following steps:

(1) weighing a proper amount of water, and placing the water in a container for later use;

(2) weighing sodium hyaluronate, sodium carboxymethylcellulose, collagen, alginate and chitosan according to specified amount;

(3) adding the components weighed in the step (2) into the container (1) containing water, standing at room temperature for 6-8 hours;

(4) stirring to uniformly dissolve the raw materials in the step (3);

(5) weighing the nonionic surfactant according to a specified amount, adding the nonionic surfactant into the mixture obtained in the step (4), and stirring for dissolving;

(6) adding water to make the concentration of each raw material reach the concentration range specified by the invention;

(7) stirring to obtain the final product.

The invention further provides application of the medical skin care dressing in preparation of a reagent for promoting transdermal absorption of a skin external medicine.

The skin external medicine comprises a medicine for treating infantile hemangioma, a medicine for treating infantile vascular malformation, a medicine for treating chloasma, a medicine for promoting local healing, or a local analgesic medicine.

The medicament for treating the infantile hemangioma comprises a beta receptor blocker, wherein the beta receptor blocker is preferably selected from one or more of timolol maleate, propranolol, carteolol hydrochloride and betaxolol hydrochloride.

The medicament for treating the infantile vascular malformation comprises sirolimus and tacrolimus.

The medicine for treating chloasma comprises tranexamic acid and levovitamin C.

The medicine for promoting local healing comprises growth factor and rehabilitation new liquid.

The local analgesic comprises lidocaine and nitroglycerin.

[ method for producing dressing of the present invention ]

The invention provides a method for preparing a medical skin care dressing, which comprises the following steps of (1) weighing a proper amount of water, and placing the water in a container for later use; (2) weighing sodium hyaluronate, sodium carboxymethylcellulose, collagen, alginate, chitosan and optional components according to a specified amount; (3) sequentially adding the components weighed in the step (2) into the container (1) containing water, and standing at room temperature for 6-8 hours; (4) stirring to uniformly dissolve the raw materials such as sodium hyaluronate, sodium carboxymethylcellulose and the like in the step (3); (5) weighing the nonionic surfactant according to a specified amount, adding the nonionic surfactant into the mixture obtained in the step (4), and stirring to dissolve the nonionic surfactant; (6) adding water to make the concentration of each raw material reach the concentration range specified by the invention; (7) stirring to obtain the medical skin care dressing.

Preparation example 1

Preparing high molecular sodium hyaluronate (purchased from Huaxi Biotechnology corporation, product code: HA-THM) with total content of 0.3g, medium molecular sodium hyaluronate (purchased from Huaxi Biotechnology corporation, product code: HA-T) with total content of 0.6g, small molecular sodium hyaluronate (purchased from Huaxi Biotechnology corporation, product code: HA-TLM) with total content of 0.6g, and sodium carboxymethylcellulose (purchased from Anhui mountain river pharmaceutic adjuvant corporation), 0.6g of collagen (purchased from West Ann Gemini Biogene technology Co., Ltd.), sodium alginate (purchased from Qingdao Huanghai biopharmaceuticals) and chitosan (purchased from Meclin biosciences) (ratio 1:1:1) were added to a vessel containing 50g of process water, and left to stand at room temperature for 6-8 hours. Then, the raw materials such as sodium hyaluronate and sodium carboxymethylcellulose are dissolved uniformly by continuous stirring with an electric stirrer, 6g of poloxamer 407 is added, the raw materials are dissolved by continuous stirring with an electric stirrer, and the total weight of the product is supplemented to 100g with production water so that the concentration of each raw material reaches the concentration specified by the invention. And (4) continuously stirring by using an electric stirrer, and uniformly mixing to obtain the product to be subpackaged. And (4) filling the obtained product into a pre-treated container according to the filling requirement by using a filling device, and sealing and capping to obtain the finished product.

Preparation example 2

The medical skin care dressing of the present invention was prepared in the same preparation method as in preparation example 1, wherein the total content of the polymeric sodium hyaluronate was 0.9g, the content of the medium molecular sodium hyaluronate was 1.8g, the content of the small molecular sodium hyaluronate was 1.8g, and the amounts of the other components added were the same as in preparation example 1.

Preparation example 3

The medical skin care dressing of the present invention was prepared in the same manner as in preparation example 1, wherein the amount of sodium carboxymethylcellulose added was 5g, and the amounts of the other ingredients added were the same as in preparation example 1.

Preparation example 4

The medical skin care dressing of the present invention was prepared according to the same preparation method as in preparation example 1, wherein the amount of poloxamer added was 1g, and the amounts of other components added were the same as in preparation example 1.

Preparation example 5

The medical skin care dressing of the present invention was prepared in the same preparation method as in preparation example 1, wherein the total addition amount of collagen, alginate and chitosan was 5g, and the addition amounts of the other ingredients were the same as in preparation example 1.

Preparation example 6

The medical skin care dressing of the present invention was prepared in the same preparation method as in preparation example 1, wherein the total content of the polymeric sodium hyaluronate was 1.8g, the content of the medium molecular sodium hyaluronate was 3.6g, the content of the small molecular sodium hyaluronate was 3.6g, and the amounts of the other components added were the same as in preparation example 1.

Preparation example 7

The medical skin care dressing of the present invention was prepared in the same manner as in preparation example 1, wherein the amount of sodium carboxymethylcellulose added was 10g, and the amounts of the other ingredients added were the same as in preparation example 1.

Preparation example 8

The medical skin care dressing of the present invention was prepared according to the same preparation method as in preparation example 1, wherein the amount of poloxamer added was 10g, and the amounts of other components added were the same as in preparation example 1.

Preparation example 9

The medical skin care dressing of the present invention was prepared in the same preparation method as in preparation example 1, wherein the total addition amount of collagen, alginate and chitosan was 10g, and the addition amounts of the other components were the same as in preparation example 1.

Performance testing

And (3) testing the sample: timolol maleate eye drops, specification: 25mg in 5mL, wherein the content of timolol is 0.5 percent, and the chemical names of the effective components are as follows: (mono) - (tert-butylamino) -3- [ (4-morpholinyl-1, 2, 5-thiadiazol-3-yl)]-2-propanol maleate (C)₁₃H₂₄N₄O₃S·C₄H₄O₄Molecular weight: 432.49, structural formula:

)

tranexamic acid injection, specification: 10ml:1.0g, approved article No.: the national Standard of medicine H200569B7, Guangzhou Baiyunshan Tianxin pharmaceutical GmbH.

Sirolimus oral solution, 50ml:50mg, Hangzhou Med Huadong pharmaceutical Co., Ltd.

First, stability test

The product of preparation example 1 was mixed with the test article in a ratio of 1:1, and the change in timolol content after mixing and standing at room temperature was examined. The test was divided into 3 groups, the first group (W1) being only samples of preparation 1, parallel with 4, 0.5g each; the second group (W2) is a mixed sample of preparation example 1 and the test sample, 5g (about 5ml) of the liquid medicine of preparation example 1 is taken, 5ml of the test sample is added and mixed evenly, and the mixture is divided into 12 parts in parallel, wherein each part is 500 mu l; the third group (W3) was test samples only, 12 aliquots of 500. mu.l each; the above 28 samples were left at room temperature for 5 days and 12 days, respectively.

Each sampling time point (5 days, 12 days): taking 2 parts of the first group of samples, diluting the first group of samples by 20 ten thousand times with methanol, and uniformly mixing the samples; taking 6 parts of the second group of samples, diluting the samples by 10 ten thousand times with methanol, and uniformly mixing the samples; and taking 6 parts of the third group of samples, diluting the samples by 20 ten thousand times with methanol, and uniformly mixing the diluted samples. On the day of analysis, samples (0 day-W1 and 0 day-W3) were freshly prepared according to the methods of group W1 and group W3, diluted 20 ten thousand times with methanol and mixed. On the day of analysis, a set of timolol standard curves was prepared. And analyzing the content of timolol in the uniformly mixed sample by adopting an LC-MS method.

TABLE 1

The test results are shown in tables 2 and 3 below:

TABLE 2 stability data-5 days

Note: the mean and relative standard deviation are statistics without day 0 data;

stability% -mean timolol content/timolol content 0 day × 100%

TABLE 3 stability data-12 days

stability% -mean timolol content/timolol content 0 day x 100%

And (3) observing the room-temperature placement stability by adopting the ratio of the average timolol content of the samples placed at room temperature to the timolol content of the samples of the same type prepared on the same day. Stability test data show that after the sample of the preparation example 1 and the test sample are mixed in equal proportion, the stability of the mixture placed at room temperature is 93.7-109.8%; the stability of the test sample placed at room temperature is 100.3%. The sample of preparation example 1 had no effect on the room temperature stability of the sample, and the sample of preparation example 1 and the sample did not undergo a chemical reaction.

Second, cytotoxicity test

This assay tested the samples of preparative example 1 for potential cytotoxic effects using mammalian L-929 cells cultured in vitro as per the requirements of the method of GB/T16886.5-2017.

The sample of preparation example 1 and the control sample were each placed in MEM medium containing 10% fetal bovine serum and extracted at 37 ℃ for 24 hours. 96-well plates (10) incubated for 24 hours after leaching was complete⁴One/well) cell culture medium was removed, replaced with corresponding extract, and placed in a cell culture chamber (37 ℃, 5% CO)₂，>90% humidity) for 24 hours. And observing the cell morphology and the cell lysis condition under the microscope after the culture is finished, and determining the cytotoxicity value of the test sample by adopting an MTT method.

The results showed that the cells in the blank control group (MEM containing 10% FBS, manufacturer: Hyclone) and the negative control group (high density polyethylene, manufacturer: Hatano Research institute. FDSC) were morphologically intact throughout the experiment and showed no cytotoxic reaction. A severe cytotoxic response was shown in the positive control group (ZDEC, manufacturer: Sigma-Aldrich). The 100% strength leaching solution of the test sample was essentially intact in cell morphology after 24 hours of incubation of the cells (Table 4), the cell viability value was 82.0% (Table 5), and the test substance was not potentially cytotoxic to L-929 cells.

TABLE 4 Observation of cell morphology

TABLE 5 MTT results

Skin irritation test

The test is carried out by adopting a New Zealand white rabbit according to the method requirement of GB/T16886.5-2017, and the potential of the sample prepared in the preparation 1 to generate skin irritation reaction under the test condition is evaluated.

In this experiment, the samples were extracted using 0.9% sodium chloride injection, shaking table extraction was performed at constant temperature of 60rpm for 72 hours at 37 ℃. Negative control solutions (sodium chloride injection, manufacturer: Shijiazhuang Siyao Co., Ltd.) were prepared under the same conditions. Gauze (about 2.5 cm. times.2.5 cm) saturated with the leach solution of the test and control samples was placed in direct contact with the skin on both sides of the spine of the animal (shaved 24 hours prior to contact) and then applied securely with a bandage for at least 4 hours. And taking down the patch after finishing. After 1 hour, 24 hours, 48 hours and 72 hours, the application site and surrounding skin tissue reactions including erythema, edema and necrosis, etc. were observed and the scores were recorded.

The results show (table 6) that animals in the negative control group (0.9% sodium chloride injection) had intact skin and no erythema, edema and necrosis of the skin during the experiment. The positive control group (SDS, manufacturer: SIGMA) animals showed marked irritation reactions such as erythema, edema, and necrosis on their skin. The animals of the leaching solution group of the test sample (preparation example 1) had intact skin and no erythema, edema and necrosis. All groups of data meet the acceptance criteria, and the test result is effective. Based on the above results, it can be concluded that: under the present experimental conditions, the test samples had no skin irritation in the irritation experiment.

TABLE 6 Observation of skin reaction results

Fourth, sensitization test

The potential skin sensitization of the samples of preparation example 1 was observed using a guinea pig maximum test method in accordance with the method requirements of GB/T16886.5-2017.

In this experiment, the samples were extracted using 0.9% sodium chloride injection and incubated at 37 ℃ for 72 hours on a constant temperature shaker at 60 rpm. Negative control solutions were prepared under the same conditions. Mixing the prepared leaching solution with Freund's complete adjuvant to obtain stable emulsifier, and injecting the emulsifier into the inner side of scapula with hair removed to perform intradermal induction and local induction. Challenge tests were performed 14 days after local induction at the non-test sites during the induction phase. The skin reactions at the challenge sites of the test and control animals were observed under full spectrum light at 24 hours and 48 hours after challenge, and the skin erythema and edema responses at each challenge site were scored according to Magnusson and Kligman grading standards.

The results (Table 7) show that the animals in the negative control group (0.9% sodium chloride injection, manufacturer: Shijiazhuang Siyao Co., Ltd.) had intact skin and no skin erythema and edema reaction during the experiment. Animals of the positive control group (2, 4-dinitrochlorobenzene (DNCB, manufacturer: TOKYO CHEMICAL INDUSTRY CO., LTD)) developed marked skin erythema and edema reactions. The test sample group (preparation example 1) leaching solution group animals had intact skin and no skin erythema and edema reaction. All groups of data meet the acceptance criteria, and the test result is effective. Based on the above results, it can be concluded that: under the present experimental conditions, the test sample has no skin sensitization in the sensitization experiment.

TABLE 7 Guinea pig sensitization skin reactions

Fifth, percutaneous absorption test

(1) The percutaneous absorption effect of the medical skin care dressing on the timolol maleate eye drops is investigated

Preparing animal skin: taking 1 male SD rat, removing skin surface villi, and adopting CO₂Euthanasia, use scalpel to remove the skin on the back of rat, and use scissors to clean the fat on the inner side of skin for transdermal absorption test.

Transdermal test, using 0.9% normal saline as absorption liquid, putting a magnetic stirrer into each diffusion cell and filling the absorption liquid (about 10 mL); covering the skin of a rat on the opening part of the diffusion cell, covering the upper sample adding groove on the skin, and fixing the diffusion cell, the skin of the rat and the sample adding groove together by using a clamp: filling water in a water tank of the percutaneous absorption instrument, and placing a diffusion cell covering the skin of a rat into a sample hole of the percutaneous absorption instrument; setting the water bath temperature to 32 ℃, setting the magnetic stirring speed to 350 r/min, and starting the instrument; after the temperature of the instrument is stable, adding 2mL of sample solution into the sample adding groove; after the addition of the sample solution, 500. mu.L of the absorbent solution was taken out from the middle of the diffusion cell at a time of 30 hours from the time when the previously determined transmittance was stable, diluted with an equal amount of methanol, and subjected to LC-MS analysis. On the day of analysis, a set of standard curve solutions of phamologel was prepared and injected together.

Preparing a sample solution: samples were set as 3 groups, the first group (G1) was a blank group, and 16mL of 0.9% physiological saline solution was used in parallel with 2 portions; the second group (G2) was a test group, and 8G (about 8mL) of the sample of preparation example 1 was added to 8mL of the sample to be tested and mixed, and used as a second group sample solution, and 6 parts were used in parallel; the third group (G3) was a control group, and 8mL of 0.5% timolol maleate eye drops were added to 8mL of 0.9% physiological saline and mixed to prepare a third group sample solution, which was divided into 6 portions in parallel.

Sample analysis

An appropriate amount of timolol maleate standard substance is accurately weighed, dissolved by methanol and subjected to constant volume, so as to prepare a stock solution of 1.0 mg/mL. Timolol maleate stock solutions were diluted with acetonitrile into standard curve samples 10ng/mL (LLOQ), 20ng/mL, 50ng/mL, 100ng/mL, 200 ng/mL. Since the substance detected by LC-MS method is timolol, the standard curve sample concentration is converted into timolol mass concentrations of 7.31ng/mL (LLOQ), 14.6ng/mL, 36.5ng/mL, 73.1ng/mL and 146 ng/mL.

The standard curve contains 5 non-zero points and the accuracy of LLOQ should be between 80-120%: besides LLOQ, the accuracy of each point on the standard curve should be between 85-115%, and the correlation coefficient (r) of the standard curve is ≧ 0.98.

Data processing and statistical analysis

Precision (relative standard deviation RSD): RSD [ ∑ (R-R) ]_Average)²/(n-1)]^1/2/R_Average×l00

Accuracy: acu% ═ C_{Determination of concentration}/C_{Theoretical concentration}×l00

Test results

TABLE 8 in vitro transdermal absorption test

Note: the concentration of the receiving solution is the measured concentration x 2 (dilution factor); the italic black data is abnormal data, and is not involved in statistics of in vitro transdermal absorption test by using SD male rat skin, the influence of the medical skin care dressing on the transdermal absorption of timolol maleate is investigated, and the result is shown in the attached table 8. Mixing timolol maleate and normal saline in equal proportion, and after transdermal absorption for 30 hours, ensuring that the content of timolol in a receiving solution is less than 7.31ng/mL of minimum limit of quantitation (LLOQ); after the sample of preparation example 1 of the present invention was transdermally absorbed for 30 hours, the mean timolol content of the receiving solution was 49.7ng/mL, and the relative standard deviation was 44.9%.

Conclusion of the experiment

The data of the percutaneous absorption test show that the percutaneous absorption rate is very low and almost not penetrated under the condition of only the timolol maleate; after the medical skin care dressing disclosed by the invention is adopted, the concentration of timolol in the receiving liquid is greatly increased, so that the medical skin care dressing disclosed by the invention can promote the transdermal absorption of timolol maleate.

The samples of preparation examples 2 to 9 were tested according to the same test method as above, and the results showed that they all had stability, no cytotoxicity, no skin irritation, no sensitization, and improved transdermal absorption rate of timolol maleate.

(2) The experimental principle of the transdermal absorption effect of the medical skin care dressing on tranexamic acid injection is examined: franze diffusion cell principle.

Grouping samples:

group A: physiological saline and tranexamic acid injection

Taking 1 tranexamic acid injection, transferring 10ml of the drug into a mixing bottle, taking 5ml of physiological saline by a pipette, putting the physiological saline into the mixing bottle, and shaking up and down for 1 minute to uniformly mix the drug and the physiological saline serving as a dressing.

Group B: the invention relates to a medical skin care dressing and tranexamic acid injection

Taking 1-count tranexamic acid injection, transferring 10ml of the medicine into a mixing bottle, pushing 5g of the medical skin care dressing prepared in preparation example 8 into the mixing bottle, shaking up and down for 1 minute to uniformly mix the medicine and the dressing, and standing for 30min until all air bubbles in the mixture disappear.

Time points for experimental investigation:

the medical skin care dressing combines the tranexamic acid injection and the tranexamic acid injection which is used independently, and 1ml of samples in the diffusion cell are received at 7 time points of transdermal 1h, transdermal 2h, transdermal 4h, transdermal 6h, transdermal 8h, transdermal 12h and transdermal 24h for subsequent analysis.

The analysis method comprises the following steps:

reagent: as shown in Table 9, all solvents and reagents were of chromatographic grade, and tranexamic acid standard was purchased from Shanghai-derived PhylloBiotech Ltd.

The instrument comprises the following steps: as shown in table 10.

TABLE 9 Experimental reagents

TABLE 10 Experimental instruments

Sample treatment: 1) centrifuging at the rotating speed of 12000r/min for 10 min; 2) the supernatant was taken out of the lined tube and checked by HPLC.

HPLC analysis:

the instrument used in this experiment was a waters 2695 model hplc equipped with a waters 2424 evaporation photodetector. The column was Diamonsil C18(2) (150 x 4.6mm,5 μm), the mobile phase was acetonitrile: 0.3% TFA water/7: 93, drift tube temperature 85 deg.C, atomizer temperature 30 deg.C, nitrogen flow rate 25psi, gain 100, mobile phase flow rate 1mL/min, column temperature 35 deg.C, sample size 10 μ L.

The standard curve of tranexamic acid standard is shown in FIG. 1, and the chromatogram is shown in FIG. 2.

TABLE 11

The experimental results are as follows:

TABLE 12 in vitro transdermal absorption test

Note: sample name 01-07 is 7 sampling time points.

And (4) conclusion: as shown in Table 12, the transdermal amounts of the drugs were higher in the group B than in the group A at the same sampling point, and it can be seen that the medical skin care dressing of the present invention has a significant penetration promoting effect on tranexamic acid, and the results are shown in FIGS. 3 to 16.

(3) The percutaneous absorption effect of the medical skin care dressing on the sirolimus oral liquid is investigated

The experimental principle is as follows: franze diffusion cell principle.

Grouping samples:

group E: CMC-Na & sirolimus oral liquid

Taking 5ml of sirolimus oral liquid by a pipette, putting the sirolimus oral liquid into a beaker, pushing 5g of CMC-Na dressing into the beaker, and stirring by a stirring rod to uniformly mix the medicine and the dressing into ointment. (particularly, it is suggested that the preparation is refrigerated in a dark place after the preparation).

And group F: the invention relates to a medical skin care dressing and sirolimus oral liquid

And (3) taking 5ml of sirolimus oral liquid by using a pipette, putting 5g of the medical skin care dressing prepared in the preparation example 8 into a beaker, and stirring by using a stirring rod to uniformly mix the medicine and the dressing into ointment. (particularly, it is suggested that the preparation is refrigerated in a dark place after the preparation).

Time points for experimental investigation:

the medical skin care dressing combines the sirolimus oral liquid and the blank CMC-Na gel combines the sirolimus oral liquid, and 1ml of samples in the diffusion cell are received at 7 time points of transdermal 1h, transdermal 2h, transdermal 4h, transdermal 6h, transdermal 8h, transdermal 12h and transdermal 24h for subsequent analysis.

The analysis method comprises the following steps:

reagent: as shown in Table 13, all solvents and reagents were of chromatographic grade and the standards were purchased from Shanghai-derived leaf Biotech Co.

The instrument comprises: as shown in table 14.

TABLE 13 Experimental reagents

TABLE 14 Experimental instruments

HPLC analysis:

the instrument used in the experiment is a waters 2695 type high performance liquid chromatograph equipped with a DAD detector. The chromatographic column was Diamonsil C18(2) (150 × 4.6mm,5 μm), the mobile phase was acetonitrile-methanol-water isocratic elution (7.5:62.5:30), the detection wavelength was 276nm, the flow rate was 1mL/min, the column temperature was 50 ℃ and the sample size was 10 μ L.

The standard curve of sirolimus standard is shown in FIG. 17, and the chromatogram is shown in FIG. 18.

Watch 15

The experimental results are as follows:

TABLE 16 in vitro transdermal absorption test

Sample name	Peak area	Concentration (ug/ml)	Contrast value
				E01	NA	NA		100％
E02	2365	0.42	100％
				E03	17354	1.26	100％
E04	21836	1.51	100％
				E05	24876	1.68	100％
E06	35513	2.27	100％
				E07	49181	3.04	100％
F01	1462	0.37	88％
				F02	6006	0.62	49％
F03	7207	0.69	46％
				F04	11326	0.92	55％
F05	37756	2.40	143％
				F06	64941	3.92	173％
F07	124714	7.27	239％

Note: sample name 01-07 is 7 sampling time points.

And (4) conclusion: as shown in Table 16, the skin penetration of the F group was slightly lower than that of the E group at the first 4 sampling points (1h, 2h, 4h and 6h), because the sustained release function of the product was observed, the skin penetration of the F group was greatly increased at the 5 th to 7 th sampling points, and the skin absorption capacity of the F group was 2.39 times that of the E group at 24 h. Since sirolimus is relatively irritating to the skin, the dosage of sirolimus is 1-2 times per day, and the slow-release and penetration-promoting effects of group F are consistent with the product characteristics, and the results are shown in FIGS. 19-32.

(4) Examine the influence of different contents of nonionic surfactant in the medical skin care dressing on the transdermal absorption

The experimental principle is as follows: franze diffusion cell principle.

Grouping samples:

group I: the content of poloxamer in the dressing of preparation example 8 was changed to 5% & tranexamic acid injection: taking 1-tranexamic acid injection, transferring 10ml of the medicine into a mixing bottle, then pushing 5g of the medical skin care dressing containing 5% of poloxamer into the mixing bottle, shaking up and down for 1 minute to uniformly mix the medicine and the dressing, and standing for 30min until all air bubbles in the mixture disappear.

And (II) group: the content of poloxamer in the dressing of preparation example 8 was changed to 7% & tranexamic acid injection: the preparation method is the same as that of the group I samples.

Group III: the content of poloxamer in the dressing of preparation example 8 was changed to 9% & tranexamic acid injection: the preparation method is the same as that of the group I samples.

Time points for experimental investigation:

the samples in the above groups are subjected to transdermal 1h, 4h and 12h at 3 time points, and 1ml of the sample in the diffusion cell is received for subsequent analysis.

The analysis method comprises the following steps: same as in experiment (2).

TABLE 17 in vitro transdermal absorption assay

Sample name	Peak area	Concentration (mg/ml)	Contrast value
				ⅠA01	1878	0.0571	100％
ⅠA02	76382	0.0943	100％
				ⅠA03	719737	0.4160	100％
ⅡB01	6782	0.0595	104％
				ⅡB02	191671	0.1520	161％
ⅡB03	1275459	0.6939	167％
				ⅢC01	454657	0.2835	496％
ⅢC02	1327373	0.7198	173％
				ⅢC03	3013105	1.5627	376％

Note: sample name 01-03 is 3 sampling time points.

And (4) conclusion: as shown in table 17, as the addition amount of poloxamer, which has a main permeation enhancing effect, in the formulation increases, the drug permeation amount at each sampling point tends to be greatly increased, and it can be seen that the permeation enhancing efficiency is directly proportional to the addition amount of poloxamer within a certain range, and the results are shown in fig. 33 to 41.

Sixth, moisture retention test

1. Principle of testing

The capacitance method is used for measuring the moisture content of the human skin stratum corneum, and is based on the obvious difference of dielectric constants of water and other substances, the capacitance values of the measured skin are different according to the moisture content of the skin stratum corneum, and the parameters can represent the moisture content of the skin.

2. Detecting an environment

The test environment of the test is at the temperature of 20-22 ℃; the relative humidity is 45.0-55.0%, and the test environmental requirements of QB/T4256-2011 cosmetic moisturizing efficacy evaluation guideline are met.

3. Detection instrument

Coriometer CM825 probe for Courage + Khazaka skin moisture test

The instrument adopts a capacitance method to measure the moisture content of the stratum corneum of the skin, the test area is the inner side of the forearm, and an average value is taken after each area is measured for 3 times. The larger the value measured by the instrument is, the higher the moisture content of the stratum corneum is.

4. Test method

According to QB/T4256-. The moisture content of the stratum corneum in the medial forearm test area was tested before and after a single use of the test product.

5. The samples were divided into three groups:

group A: examining the moisturizing effect of the gel prepared by mixing 5ml of the medical skin care dressing prepared in the preparation example 1 and 5ml of purified water on skin;

group B: examining the moisturizing effect of the gel prepared by mixing 5ml of the medical skin care dressing and 10ml of purified water in the preparation example 1 on the skin;

group C: the skin moisturizing effect of the medical skin care dressing of preparation example 1 was examined by spraying 5ml of purified water onto the skin of a gel prepared by mixing 5ml of the dressing with 10ml of purified water.

6. Experimental time points:

the moisture content of the skin after 0, 2,4 and 8 hours of gel application is respectively tested, 10 persons in each group are taken as an average value.

7. Results of the experiment

Watch 18

And (4) conclusion:

(1) the highest percentage of skin moisture increase occurs in the group with the lowest moisture content in the skin (i.e., age group 45-55 years), indicating that the product has better skin moisturization performance in the more water deficient state. However, the moisturizing time in the water-deficient state is not long enough, and the administration frequency needs to be increased.

(2) The moisturizing effect is gradually reduced along with the prolonging of the using time, the level is recovered to the level before the use after 8 hours, the maximum time interval of the administration time is not more than 8 hours, and the administration times are preferably 4-6 times per day.

(3) The higher the concentration of the product used by the subjects in the same age group, the better the moisturizing effect, which indicates that the external water amount is not the source of the moisturizing performance of the product, and the moisturizing mechanism of the product is derived from improving the skin cell permeability, but not from the external water source.

Seventh, film Forming Property experiment

1. Formulation components

The formula A is as follows: the medical skin care dressing in which the poloxamer content in preparation example 8 was changed to 5%; and the formula B is as follows: the medical skin care dressing in which the poloxamer content in preparation example 8 was changed to 7%; and a formula C: the content of poloxamer in the dressing of preparation example 8 was changed to 9% of that of the medical skin-care dressing.

2. Environment(s)

Temperature: room temperature (25 ℃. + -. 2 ℃); humidity: 60% +/-10%.

3. Efficiency of film formation

As shown in fig. 42-44.

4. Conclusion

The film forming time of the formula C (namely the poloxamer content in the formula is 9%) is shortest, and the film forming time of the formula B is longest, so that the film forming time is inversely proportional to the addition amount of the poloxamer within a certain addition amount range.

The medical skin care dressing provided by the invention can be uniformly coated on the skin for 20-30 min to form a film quickly, and the film is not adhered to clothes after being formed, so that the daily work and life of a patient are not influenced. The timolol eye drops are eye pressure lowering medicines, and serious adverse reactions can occur when the timolol eye drops flow into eyes of children patients if the timolol eye drops cannot be fixed, so that the product has the advantage especially at special diseased parts such as eye sockets, ear margins and perineum parts.

Eighthly, the medical skin care dressing provided by the invention has a treatment effect on infantile hemangioma by combining with timolol maleate eye drops

The experimental inclusion criteria are as follows:

superficial infantile hemangiomas were diagnosed according to the hemangioma classification method of Waner et al:

1. the tumor thickness is less than or equal to 3 mm; the age is less than 6 months;

2. has not received other related treatments;

3. the maximum diameter of hemangioma is less than 5 cm; the surface has no ulceration, infection, bleeding and the like;

4. eliminating congenital heart disease, bronchial asthma, etc.

Experimental exclusion criteria:

1. the age is more than 6 months;

2. the treatment of laser, freezing, local injection and the like is performed before;

3. the tumor body has regression recently or has regression signs;

4. the tumor thickness is more than 3 mm.

The experimental method comprises the following steps:

40 patients with superficial infantile hemangioma are selected as the study object and randomly divided into 20 cases of the experimental group and the control group according to different treatment methods.

Experimental groups: 0.5% timolol maleate eye drops (Wuhan Wujing pharmaceutical Co., Ltd., national standard H42021078) are uniformly mixed with 5g of the medical skin care dressing obtained in preparation example 8 to form a gel, and the gel is uniformly applied to the surface of hemangioma 3 times a day for 6 months continuously.

Control group: the medicine cotton ball is wetted by 0.5% timolol maleate eye drops and is uniformly applied on the surface of hemangioma for 20min, 3 times a day and 6 months continuously.

And (3) recording an experiment:

observing the changes of hemangioma color, size and texture of two groups of patients;

observing whether the skin around the affected part has the adverse reactions such as ulceration, rash, dryness, desquamation and the like;

and observing the consumption amount of the timolol maleate eye drops.

The color, texture and size of hemangioma, whether dry desquamation lesion exists on the skin around the affected part, and the using amount of the timolol maleate eye drops are observed and recorded.

Judging the treatment effect:

the treatment effect is divided by adopting four-level standard proposed by Achauer et al:

watch 19

Grade	Criterion of evaluation
		Difference (D)	The tumor size is reduced by less than 25%
In (1)	The skin color fading and tumor body are not less than 26%
		Good taste	The color of the skin damage is faded and the tumor body is not lower than 51 percent;
superior food	Preferably, the skin loss is more than 75 percent, and the tumor body does not see the disease damage part and recovers or has little difference with the normal skin color

The effective rate is calculated to be equal to or more than moderate, the cure rate is calculated to be superior, and the difference is regarded as ineffective.

Table 20 (experimental groups a001 to a020, control groups B001 to B020):

and (4) conclusion: the effective rate of the experimental group is 95 percent, and the effective rate of the control group is 90 percent; the cure rate of the experimental group is 75 percent, and the cure rate of the control group is 45 percent; the experimental group has no adverse reaction, and the control group has 25% of adverse reaction; the average dosage of the drugs in the experimental group was 18 bottles, and the average dosage of the drugs in the control group was 43 bottles.

Therefore, the medical skin care dressing disclosed by the invention is combined with timolol maleate, so that a good treatment effect on infantile hemangioma can be realized. The medical grade sodium hyaluronate with multiple molecular weights in the dressing can be deeply moisturized and integrally treated and repaired, and the children patients do not have the side reaction symptoms of medicines such as red swelling, peeling, desquamation, ulceration and the like.

Ninth, the medical skin care dressing of the invention combines with tranexamic acid to treat chloasma

Experimental drugs: taking 1-count tranexamic acid injection, transferring 10ml of the medicine into a mixing bottle, pushing 5g of the medical skin care dressing prepared in preparation example 8 into the mixing bottle, shaking up and down for 1 minute to uniformly mix the medicine and the dressing, and standing for 30min until all air bubbles in the mixture disappear.

Control drugs: tranexamic acid injection.

Grouping experiments:

experimental groups: the experimental medicine is used for 11 chloasma patients;

control group: the control drug was used for 14 chloasma patients.

Change of chloasma area before and after treatment of two groups of subjects

The change in the chloasma area was counted at 2 weeks (2W), 4 weeks (4W), 8 weeks (8W) and 12 weeks (12W) after the administration, and the results are shown below:

TABLE 21

TABLE 22

And (4) conclusion: the external application of the medical skin care dressing of the invention and the treatment scheme of tranexamic acid is superior to the treatment scheme of single external application of tranexamic acid injection in the aspect of treating chloasma.

Industrial applicability

The medical skin care dressing provided by the invention is a clinical skin medicament, can be used in combination with a medicament for treating infantile hemangioma, a medicament for treating infantile vascular malformation, a medicament for treating chloasma, a medicament for promoting local healing, or a local analgesic medicament, particularly provides a convenient administration mode for the medicament for treating infantile hemangioma, improves the transdermal absorption rate of the medicament, and has the effects of moisturizing, repairing and promoting healing of an affected part.

Claims

1. A medical skin care dressing characterized by comprising a nonionic surfactant at a concentration of not less than 1 wt%, sodium hyaluronate at a concentration of not less than 1 wt%, sodium carboxymethylcellulose at a concentration of not less than 0.1 wt%, at least one selected from the group consisting of collagen, alginate and chitosan at a total concentration of not less than 0.1 wt%, and water, with respect to the entirety of the medical skin care dressing.

2. A skin care dressing according to claim 1, characterized in that the non-ionic surfactant is a polyoxyethylene type non-ionic surfactant, preferably the non-ionic surfactant comprises a poloxamer.

3. The skin care dressing of claim 1 or 2, wherein the sodium hyaluronate comprises a polymeric sodium hyaluronate, a medium molecular sodium hyaluronate, and a small molecular sodium hyaluronate,

4. The skin care dressing of claim 3, wherein the molecular weight of the polymeric sodium hyaluronate is 1800kDa, the molecular weight of the medium molecular sodium hyaluronate is 1000-1800kDa, and the molecular weight of the small molecular sodium hyaluronate is 10-1000 kDa.

5. A medical skin care dressing according to any one of claims 1 to 4, wherein: the concentration of the nonionic surfactant is 1 to 10 wt%, preferably 2 to 8 wt%, more preferably 3 to 6 wt%,

the concentration of the sodium hyaluronate is 1-10 wt%, preferably 1-5 wt%, more preferably 1-3 wt%,

the concentration of the sodium carboxymethylcellulose is 0.1-10 wt%, preferably 0.5-2 wt%, more preferably 0.5-1 wt%,

the total concentration of at least one selected from the group consisting of collagen, alginate and chitosan is 0.1% to 10% by weight, preferably 0.4 to 4% by weight, more preferably 0.5 to 2% by weight.

6. A method of making a medical skin care dressing according to any one of claims 1 to 5, characterized in that: the method comprises the following steps:

(4) stirring to uniformly dissolve the raw materials in the step (3);

(7) stirring to obtain the final product.

7. Use of a medical skin care dressing according to any one of claims 1 to 5 in the manufacture of an agent for promoting transdermal absorption of a drug for external application to the skin.

8. Use according to claim 7, characterized in that: the skin external medicine comprises a medicine for treating infantile hemangioma, a medicine for treating infantile vascular malformation, a medicine for treating chloasma, a medicine for promoting local healing, or a local analgesic medicine.

9. The use according to claim 8, the medicament for treating pediatric hemangiomas comprises a beta blocker, preferably selected from one or more of timolol maleate, propranolol, carteolol hydrochloride, betaxolol hydrochloride,

the medicine for treating the infantile vascular malformation comprises sirolimus and tacrolimus,

the medicine for treating chloasma comprises tranexamic acid and levorotatory vitamin C,

the medicine for promoting the local healing comprises growth factors and new rehabilitation liquid,

the local analgesic comprises lidocaine and nitroglycerin.