CN114886938A

CN114886938A - Composite composition suitable for psoriasis and preparation method thereof

Info

Publication number: CN114886938A
Application number: CN202210483829.6A
Authority: CN
Inventors: 不公告发明人
Original assignee: Niten Guangzhou Biotechnology Co ltd
Current assignee: Niten Guangzhou Biotechnology Co ltd
Priority date: 2022-05-05
Filing date: 2022-05-05
Publication date: 2022-08-12
Anticipated expiration: 2042-05-05
Also published as: CN114886938B

Abstract

The invention discloses a composite composition suitable for psoriasis and a preparation method thereof, belonging to the technical field of medicines. The composition is a pure oil ointment with the functions of epidermolysis bacteria prebiotics and penetration barrier, which is formed by combining caprylic/capric triglyceride, perilla seed oil, ascorbic acid ester, cholesterol, white beeswax, ceramide, hydrogenated lecithin and squalane. After the psoriasis patient uses the skin care product, a layer of stable solid grease film can be formed, the physical barrier function of skin permeation is improved, the abundance of beneficial bacteria such as staphylococcus epidermidis, propionibacterium, malassezia and the like in a skin lesion area is obviously increased, the abundance of pathogenic bacteria such as staphylococcus aureus, streptococcus, candida and the like is reduced, the microecology of the skin surface is recovered, the adaptive immune response of the skin is improved, scales and pruritus are obviously reduced, and the severity of psoriasis is reduced. The product is solid at normal temperature, is easy to spread and coat on the surface of skin to form a film, and avoids liquefaction into oil due to body temperature.

Description

Composite composition suitable for psoriasis and preparation method thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a composite composition suitable for psoriasis and a preparation method thereof.

Background

Psoriasis is a chronic inflammatory disease, has a long course of disease and is easy to relapse, and brings adverse effects to the physical health and mental conditions of patients. According to the statistics of the world health organization, about 1.25 million people worldwide suffer from psoriasis, but the causes of the psoriasis are not clearly researched.

At present, no method for completely curing psoriasis exists, and the disease condition can be effectively controlled and the life quality of patients can be improved by selecting proper external medicines, physical therapy or systemic medication and other means. According to the psoriasis diagnosis and treatment guide, the external medicine can be independently adopted for treating the localized psoriasis with skin damage less than 3 percent of the body surface area; clinically, 80% of patients with psoriasis belong to mild psoriasis, and the external medicine treatment can relieve the illness state, and in addition, 20% of patients with psoriasis are moderate psoriasis and severe psoriasis and usually need combined treatment. Therefore, the external medicine plays a significant role in the treatment of psoriasis.

The currently marketed external medicines mainly comprise glucocorticoids, coal tar oils, retinoic acid compounds, vitamin D3 analogs, calcineurin inhibitors, heat-clearing traditional Chinese medicines and the like. The medicines mainly play roles in resisting inflammation, inhibiting immune reaction, loosening cutin and the like, have certain curative effect, are easy to relapse, have more adverse reactions after long-term use, and urgently need a brand new medicine which is safer and has definite curative effect to be put on the market in clinic.

At present, more and more researches show that the surface symbiotic bacteria of the psoriasis lesion area are disordered, the abundance of staphylococcus aureus, streptococcus and candida is increased, the abundance of staphylococcus epidermidis, propionibacterium and malassezia is obviously reduced, and the staphylococcus aureus, streptococcus and candida have superantigens, can induce and strongly induce Th17 reaction, generate cytokines such as IL-17A, IL-17F and IL22 and the like, cause abnormal T cell immune reaction of skin and cause skin lesion and abnormal keratinization. Beneficial bacteria such as Staphylococcus epidermidis, Propionibacterium, and Ralsbad bacteria can decompose triglyceride in sebum to produce short chain fatty acid and other bacteria metabolite, and regulate CD ₄ ⁺ T cells maintain skin adaptive immune homeostasis. Additional studies have found that decreased sebaceous gland content in plaques of psoriasis is an important factor in the bacteria responsible for skin commensal bacterial disorders.

The decrease in sebum, keratosis insufficiency and keratolysis in the skin lesion area of psoriasis patients directly leads to the destruction of the skin epidermal permeation barrier. Abnormal epidermal permeability barrier leads to abnormal proliferation and differentiation of keratinocytes, resulting in parakeratosis; the abnormal permeation barrier can also cause the loss of skin moisture or the permeation of moisture and foreign matters in the environment, the keratin hydration is caused by the permeation of the moisture, and the foreign matter blocking function is reduced; the abnormal permeability barrier causes antigen invasion, promotes the migration of skin immune cells, provides convenience for the migration and aggravates immune response. Many factors are mixed, and the damage of the permeability barrier enters the vicious circle, which is finally shown as thickening and drying of the skin and difficult recovery of the disease.

However, no drug for improving skin epidermal symbiotic bacteria and improving the permeability barrier is on the market at present.

Disclosure of Invention

In order to solve the technical problems that the prior art cannot improve the abnormal conditions of 'skin symbiotic bacteria disorder', 'epidermal penetration barrier damage' and the like of psoriasis patients and has poor healing effect, the invention aims to provide a composite composition suitable for psoriasis and a preparation method thereof.

The technical scheme adopted by the invention is as follows: a composite composition suitable for psoriasis is prepared by mixing prebiotic oil and fat, permeation barrier oil and melting point regulating oil;

the prebiotic oil is prepared from the following raw materials in parts by mass: 39-60 parts of caprylic/capric triglyceride, 0.4-4 parts of perilla seed oil and 3-15 parts of ascorbic acid ester;

the permeation barrier grease is prepared from the following raw materials in parts by mass: 7-15 parts of cholesterol, 19-36 parts of white beeswax, 0.1-1 part of ceramide and 0.1-2 parts of hydrogenated lecithin;

the melting point adjusting grease is squalane 2-9.5 parts.

Preferably, the prebiotic oil and fat is prepared from the following raw materials in parts by mass: 45-52 parts of caprylic/capric triglyceride, 1.5-3 parts of perilla seed oil and 5-13 parts of ascorbic acid ester;

preferably 48 parts of caprylic/capric triglyceride, 2.9 parts of perilla seed oil and 8 parts of ascorbyl ester.

Preferably, the permeation barrier grease is prepared from the following raw materials in parts by mass: 9-12 parts of cholesterol, 23.5-31 parts of white beeswax, 0.4-0.9 part of ceramide and 0.5-1.7 part of hydrogenated lecithin;

preferably, the permeation barrier grease is prepared from the following raw materials in parts by mass: 10 parts of cholesterol, 26 parts of white beeswax, 0.5 part of ceramide and 1.2 parts of hydrogenated lecithin.

Preferably, the melting point adjusting grease is 4-9 parts of squalane;

preferably, the melting point adjusting oil is squalane 6 parts.

Preferably, the ascorbyl ester comprises one or more of ascorbyl palmitate, ascorbyl dipalmitate, ascorbyl methylsilanol pectate, ascorbyl tocopherol maleate, ascorbyl tetraisopalmitate, ascorbyl stearate, tetrahexyldecyl ascorbate, aminopropanol ascorbyl phosphate;

ascorbyl palmitate is preferred.

Preferably, the ceramide is ceramide phosphorylcholine and/or ceramide phosphorylethanolamine.

Preferably, the hydrogenated lecithin is soybean hydrogenated lecithin.

A method of preparing a composite composition suitable for psoriasis, the method comprising:

weighing caprylic/capric triglyceride, perilla seed oil, ascorbic acid ester, cholesterol, white beeswax, ceramide, hydrogenated lecithin and squalane according to corresponding mass fractions, mixing, stirring and cooling.

Preferably, the mixing and stirring conditions are as follows:

the rotation speed is 400-500r/min, and the temperature is raised to 130-150 ℃, and the stirring is continued for 2-3 min.

Preferably, the mixed and stirred raw materials are cooled to 40-50 ℃.

The invention has the beneficial effects that:

the invention provides a composite composition suitable for psoriasis, which is prepared by combining caprylic/capric triglyceride, perilla seed oil, ascorbyl ester, cholesterol, white beeswax, ceramide, hydrogenated lecithin and squalane. The composition of the application is an external grease ointment preparation.

Wherein, caprylic/capric triglyceride is the main component of sebum and is also the main nutrient substance of body surface symbiotic bacteria, and perilla seed oil and ascorbyl ester have the function of inhibiting the proliferation of pathogenic bacteria and are also the nutrient substance of body surface symbiotic bacteria. The caprylic acid/capric acid triglyceride, the perilla seed oil and the ascorbyl ester are mixed to generate the optimal effect of the skin commensal bacteria prebiotics, so that the abundance of staphylococcus epidermidis, propionibacterium, lipophilic malassezia and other probiotics in psoriasis lesion areas is remarkably increased, and the abundance of staphylococcus aureus, streptococcus, candida and other harmful bacteria is reduced.

The white beewax is wax with good sealing performance, strong barrier effect, high melting point, close to the wax in the epidermis, stable chemical property, high viscosity, larger brittleness and insufficient plasticity; cholesterol is a main component of a cell membrane of an organism, has high melting point, stable chemical property, good adhesion with a skin cuticle, and has the defects of too high viscosity and poor film forming property; ceramide is a matrix between skin horny cells, has high melting point, stable chemical property and good adhesiveness with skin horny layers, and has the defects of high viscosity and poor film forming; hydrogenated lecithin is a component of cell membranes, has high melting point, chemical stability and good air permeability, and has the defects of poor sealing property and barrier property. The 4 kinds of grease are compounded, make up for deficiencies of each other, have high melting point, stability, sealing property, plasticity and skin affinity, have high adhesion with skin and cutin, are smeared on the surface of a psoriasis skin damage area, are easily filled between loose cutins, form a stable solid grease film, play the best role of permeating physical barrier (barrier function), can slow down the permeation of moisture, can also prevent the loss of the moisture in the skin and can also prevent the invasion of other foreign matters including pathogenic bacteria and other antigens.

The squalane is stable liquid oil, has high skin affinity, good gas permeability, easy spreading, no greasiness, low melting point, and no solidification below-38 deg.C. In the test construction, the ointment is easy to be smeared into a film when the physical characteristic melting point of the product mixed with the oil is in the range of 45-65 ℃, the film has good stability at the body surface temperature, is not easy to liquefy, and has good use feeling. Therefore, squalane is selected as the raw material for adjusting the melting point of the mixed grease, and the proper dosage is adopted according to the mixture ratio of other raw materials.

According to the application, a formula is established through a large number of experiments, and a pure oil ointment with the functions of epidermal symbiotic bacteria prebiotics benefiting and permeation barrier is formed. After bathing, after phototherapy, in the morning every day and before falling asleep, a proper amount of ointment is applied to the skin lesion part, a layer of stable solid grease film can be formed, the skin permeation physical barrier function is improved, the abundance of beneficial bacteria such as staphylococcus epidermidis, propionibacterium, malassezia and the like in the skin lesion area can be obviously increased, the abundance of pathogenic bacteria such as staphylococcus aureus, streptococcus, candida and the like is reduced, the skin surface microecology is restored, the skin adaptive immune response is improved, the scales and the pruritus are obviously reduced, and the severity of psoriasis is reduced.

The composition is used for solving the defects that the existing external preparation is easy to relapse and has more adverse reactions after long-term use mainly through pharmacological actions such as anti-inflammation, immune reaction inhibition, cutin loosening and the like, is specially used for solving the abnormal conditions of ' skin commensal bacteria disorder ', epidermal penetration barrier damage ' and the like existing in the skin damage part of a psoriasis patient, has the effects of improving the skin commensal bacteria steady state, improving adaptive immunity, supplementing sebum, improving the epidermal penetration barrier function and the like, can obviously reduce scales and pruritus, delays relapse and reduces the severity of psoriasis.

(IV) the product of the application belongs to a brand-new psoriasis treatment medicinal preparation, and has the following beneficial effects compared with the prior art: 1) can improve the phenomenon of 'symbiotic bacteria disorder' at the skin lesion part of psoriasis, increase the abundance of probiotics such as staphylococcus epidermidis, propionibacterium, lipophilic malassezia and the like, reduce the abundance of harmful bacteria such as staphylococcus aureus, streptococcus, candida and the like, and improve T cell immune response; 2) the grease ointment is smeared on the surface of the psoriasis skin lesion to form a stable solid grease film with good air permeability, supplements the sebum of the psoriasis skin lesion part, plays a role in penetrating physical barrier (barrier), can reduce the penetration and the loss of water, and can also prevent the invasion of other foreign matters including pathogenic bacteria and other antigens; 3) the product of the invention is remarkably different from the externally applied psoriasis medicament on the market at present in the aspects of formula raw materials and treatment mechanism, and can remarkably reduce scales and pruritus of psoriasis, delay relapse and reduce the severity of psoriasis; 4) the product does not contain chemical surfactant (emulsifier), preservative, pigment, essence and moisture. Use of emollients under dry skin conditions: consensus statement (2013): products containing emulsifiers can aggravate skin itch and reduce the skin permeability barrier; the antiseptic, pigment and essence are all used as foreign matters to aggravate the itching and symptoms of psoriasis. The water can swell the loose horny layer of the psoriasis skin, and aggravate the damage of the barrier function of the skin; 5) the mixed grease is solid at normal temperature, is easy to spread and coat on the surface of the skin to form a film, avoids being liquefied into oil due to body temperature, prevents 'greasy feeling', and adopts raw materials with higher affinity with the skin as compatible raw materials, so that the product has better use feeling than the marketed pure grease product, and achieves the effect of 'grease is not greasy'.

Drawings

FIG. 1 is a graph of experimental studies on D6 skin lesions in a psoriasis mouse;

FIG. 2 shows the pathological examination results of skin lesions of psoriasis mice in experimental example study.

Detailed Description

The present invention is further illustrated below with reference to specific examples. It will be appreciated by those skilled in the art that the following examples, which are set forth to illustrate the present invention, are intended to be part of the present invention, but not to be construed as limiting the scope of the present invention. The reagents used are all conventional products which are commercially available.

Example 1:

the formula is as follows: 45 grams of caprylic/capric triglyceride, 2.9 grams of perilla seed oil, 15 grams of ascorbyl palmitate, 7 grams of cholesterol, 19 grams of white beeswax, 0.1 gram of ceramide phosphorylcholine, 2 grams of soybean hydrogenated lecithin and 9 grams of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 2min under the condition that the temperature is gradually increased to 130 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 50 ℃, and discharging, thereby obtaining the grease ointment with the melting point of 50-55 ℃.

Example 2:

the formula is as follows: 60 grams of caprylic/capric triglyceride, 0.5 gram of perilla seed oil, 3 grams of ascorbyl dipalmitate, 15 grams of cholesterol, 19 grams of white beeswax, 0.4 gram of ceramide phosphoethanolamine, 0.1 gram of soybean hydrogenated lecithin and 2 grams of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 3min under the condition that the temperature is gradually increased to 150 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 40 ℃, and discharging, thereby obtaining the grease ointment with the melting point of 45-51 ℃.

Example 3:

the formula is as follows: 52 g of caprylic/capric triglyceride, 4 g of perilla seed oil, 5 g of ascorbyl methyl silanolate pectate, 10 g of cholesterol, 23.5 g of white beeswax, 1 g of ceramide phosphorylcholine, 0.5 g of soybean hydrogenated lecithin and 4 g of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 2min under the condition that the temperature is gradually increased to 130 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 50 ℃, and discharging, thereby obtaining the grease ointment with the melting point of 45-52 ℃.

Example 4:

the formula is as follows: 45 grams of caprylic/capric triglyceride, 4 grams of perilla seed oil, 5 grams of ascorbyl tocopherol maleate, 7 grams of cholesterol, 36 grams of white beeswax, 0.5 gram of ceramide phosphoethanolamine, 0.5 gram of soybean hydrogenated lecithin and 9.5 grams of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 3min under the condition that the temperature is gradually increased to 150 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 40 ℃, and discharging to obtain the grease ointment with the melting point of 51-56 ℃.

Example 5:

the formula is as follows: 39 g of caprylic/capric triglyceride, 0.4 g of perilla seed oil, 13 g of ascorbyl tetraisopalmitate, 10 g of cholesterol, 31 g of white beeswax, 0.9 g of ceramide phosphorylcholine, 1.7 g of soybean hydrogenated lecithin and 4 g of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 2min under the condition that the temperature is gradually increased to 130 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 50 ℃, and discharging to obtain the grease ointment with the melting point of 62-65 ℃.

Example 6:

the formula is as follows: 45 grams of caprylic/capric triglyceride, 1.5 grams of perilla seed oil, 5 grams of ascorbyl stearate, 9 grams of cholesterol, 23.5 grams of white beeswax, 0.4 gram of ceramide phosphoethanolamine, 0.5 gram of soybean hydrogenated lecithin and 4 grams of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 3min under the condition that the temperature is gradually increased to 130 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 40 ℃, and discharging to obtain the grease ointment with the melting point of 48-54 ℃.

Example 7:

the formula is as follows: 52 g of caprylic/capric triglyceride, 3 g of perilla seed oil, 13 g of tetrahexyldecanol ascorbate, 12 g of cholesterol, 31 g of white beeswax, 0.9 g of ceramide phosphorylcholine, 1.7 g of soybean hydrogenated lecithin and 9 g of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 2min under the condition that the temperature is gradually increased to 150 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 50 ℃, and discharging, thereby obtaining the grease ointment with the melting point of 53-59 ℃.

Example 8:

the formula is as follows: 48 g of caprylic/capric triglyceride, 2.9 g of perilla seed oil, 8 g of aminopropanol ascorbyl phosphate, 10 g of cholesterol, 26 g of white beeswax, 0.5 g of ceramide phosphoethanolamine, 1.2 g of soybean hydrogenated lecithin and 6 g of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into a mixing and stirring device, continuously stirring for 3min under the condition that the temperature is gradually increased to 150 ℃ at the rotation speed of 400-500r/min, then cooling and stirring to 40 ℃, and discharging, thereby obtaining the grease ointment with the melting point of 50-56 ℃.

Based on the recipe in example 3, a control example was designed

Comparative example 1:

the formula is as follows: 52 g of caprylic/capric triglyceride, 4 parts of olive oil and 5 parts of ascorbyl palmitate; 23.5 parts of white beewax, 10 parts of cholesterol, 0.5 part of hydrogenated lecithin, 1 part of ceramide and 4 parts of squalane;

weighing the raw materials according to the corresponding mass, adding the raw materials into mixing and stirring equipment, continuously stirring for 2min under the condition that the temperature is gradually increased to 130-150 ℃ at the rotating speed of 400-500r/min, and then cooling and stirring to 50 ℃, and discharging to obtain a finished product.

To verify the efficacy of perilla seed oil, the perilla seed oil of example 3 was changed to olive oil to obtain comparative example 1.

Comparative example 2:

the formula is as follows: 57 parts of caprylic/capric triglyceride and 4 parts of perilla seed oil; 10 parts of cholesterol, 23.5 parts of white bee wax, 1 part of ceramide, 0.5 part of hydrogenated lecithin and 4 parts of squalane;

To verify the efficacy of ascorbate, ascorbate in example 3 was removed to provide comparative example 2.

Comparative example 3:

the formula is as follows: 52 parts of caprylic/capric triglyceride, 4 parts of perilla seed oil and 5 parts of ascorbic acid ester; 10 parts of cholesterol, 23.5 parts of white beeswax, 1 part of ceramide and 4 parts of squalane;

In order to verify the therapeutic effect of hydrogenated lecithin, the hydrogenated lecithin material in example 3 was removed to obtain comparative example 3.

Comparative example 4:

the formula is as follows: 52 parts of caprylic/capric triglyceride, 4 parts of perilla seed oil and 5 parts of ascorbic acid ester; 10 parts of cholesterol, 23.5 parts of white beeswax, 0.5 part of hydrogenated lecithin and 4 parts of squalane;

In order to verify the efficacy of ceramide, the ceramide material in example 3 was removed to obtain comparative example 4.

Comparative example:

"A sebum-like membrane fraction and method for its preparation" in the data "example 1", as a comparative example:

the raw materials comprise the following components in percentage by weight: 60 parts of triglyceride (15 parts of sunflower seed oil, chicory seed oil, shea butter and grape seed oil respectively), 23 parts of waxes (5 parts of carnauba wax palmitate wax, 10 parts of beeswax and 8 parts of candelilla wax), 12 parts of squalane and 5 parts of dihydrocholesterol.

Weighing wax and dihydrocholesterol, premixing at 150 deg.C, stirring at rotation speed of 400-500r/min, cooling to 90 deg.C, slowly adding the premixed raw materials, stirring to 30 deg.C, and discharging to obtain paste with melting point of 12-18 deg.C, which is liquid product at 30 deg.C.

Experimental example:

the first study is as follows: evaluation of the therapeutic efficacy of Imquimod cream for psoriasis in a mouse model

An animal SPF grade C57BL/6J mouse, male, with a body mass of 20-22g, purchased from Beijing Huafukang Biotechnology GmbH, with an animal certification number of SCXK (Jing) 2014-0004, imiquimod cream (Sichuan mingxin pharmaceutical industry, Inc.); peroxidase sealing solution, DAB color development solution and fluorescent sealing tablet (China fir gold bridge); anti-CD 3 antibody (Abcam), Alexa

488 goat anti-rabbit IgG (Guanxing science and technology Limited space Co.), BCA kit (Thermo), tissue dehydrator (Leica-ASP300S), embedding machine (Leica-EG1140C) upright fluorescence microscope and image analysis system (Zeiss-IMAGERZ 2).

The method is described in the literature [ van der Fits L, Mourits S, Voerman JS, et al, Imiquormated psioriasis-like skin inflammation in mice is meditated via the IL-23/IL-17axis. J Immunol,2009,182 (9): 5836 + 5845] A classical model of psoriasis-like lesions was induced by imiquimod by intraperitoneal injection of 80mg/kg sodium pentobarbital anaesthetized and scraping of the hair on the back by about 2 cm. times.2 cm.

Randomized into 6 groups: blank control group; a model group; example 3 (experimental group); comparative example 1; comparative example 2; comparative example group. Each group contains 5 animals, and is fed in a single cage. Except for the blank control group, 62.5mg of 5% imiquimod cream was applied externally 1 time a day. After 6 days, the mice were sacrificed. The administration mode comprises the following steps: after the imiquimod cream is smeared, 50mg of the paste to be tested is smeared for 2h and 12h, and the continuous administration is carried out for 5 days.

Observation indexes are as follows:

1) skin lesion performance and PASI score in mice: the skin lesions of the mice were photographed every day, and the severity scores were made according to erythema, scaling and infiltration, and the evaluation was made from 3 aspects of erythema, infiltration and scaling, respectively, and classified into 5 grades according to severity: no point of 0, a lightness point of 1, a moderate point of 2, a severe point of 3, a very severe point of 4 and a total point of 12.

2) Detecting the water content of the epidermis of the mouse skin: before material taking, the water content of the skin damage epidermis on the back of each group of mice is detected by a water-oil pen, repeated for 3 times, and the average value is taken.

3) Pathological change detection tissues of the mouse skin lesion are fixed and dehydrated to prepare paraffin sections of 5 mu m for HE staining. And measuring the thickness of the epidermis of the picture under the 40-time objective by using a ZEN image analysis system.

4) Immunohistochemistry method for detecting infiltration of mouse skin damage CD3+ T cells:

paraffin sections were dewaxed and antigen repaired, then sealed with peroxidase-sealed chamber at 37 ℃ for 10min, incubated with 10% goat serum at 37 ℃ for 30min, and incubated overnight at 4 ℃ with loicrin (1: 50) I antibody. After incubation for 1h at 37 ℃ with goat anti-rabbit IgG antibody, color was developed. CD3+ T cell infiltration was observed under an upright optical microscope.

The research results are as follows:

1) skin lesion expression in psoriasis-like mice:

as shown in fig. 1, the placebo mice had smooth skin on the back and a reddish flesh color. With the application of the mequinmod, the skin lesions of the model group mice show psoriasis-like manifestations such as erythema, scaling, infiltration and the like.

TABLE 1 psoriasis mice skin lesion PASI score (mean)

Grouping	The next day	The third day	Day six
				Blank control	0	0	0
Model set	3	6	11
				Example 3	2	3	2*
Comparative example 1	3	4	5*△
				Comparative example 2	3	5	6*△
Comparative example	3	6	9△

Note: *: through the rank sum test, compared with a model group, the significance difference is generated (p is less than 0.05); and (delta): compared with example 3, the difference is significant by the rank sum test (p is less than 0.05).

As shown in table 1, compared with the model group, the mice of each group showed erythema, scaling, infiltration, etc. on the 2 nd day of administration; the mice in each group were progressively trended differently in the PASI score 3 days after dosing.

2) Water content and thickness of skin lesion

As shown in fig. 2, pathological observation shows that the skin of blank mice on day 6 is thin, the infiltration of dermal lymphocytes is less, the epidermis of model mice is obviously thickened (P <0.05), the epidermal cells are parakeratosis, and the dermis is infiltrated by a large amount of lymphocytes; the epidermis of the other groups was thinned and dermal lymphocyte infiltration was reduced compared to the model group; the comparative example group had a skin thickness that was thicker than the comparative examples 1-2(P <0.05), and the example 3 group had a skin thickness that was significantly lower than the comparative examples 1-2(P < 0.05).

TABLE 2 moisture content and skin damage thickness of the epidermis

Grouping	Moisture content (%) of epidermis at skin lesion site	Thickness of skin damage (mum)
			Blank control	34.65±1.02	12.34±0.25
Model set	17.24±0.89	89.36±6.41
			Example 3	33.26±0.93*	13.95±0.47*
Comparative example 1	29.74±1.59*	31.01±2.83*△
			Comparative example 2	28.38±1.16*	33.79±3.84*△
Comparative example	18.43±1.42	78.42±7.35△

Note: *: compared with the model group, the significant difference exists (p is less than 0.05); and (delta): compared with example 3, the difference is significant (p < 0.05).

As shown in table 2, the water content of the epidermis of the model group mice was significantly reduced (P <0.05) at day 6 compared to the blank control group, suggesting that the epidermal permeability barrier was impaired; the water content of the epidermis was increased in all the other groups of mice compared with the model group.

3) T lymphocyte activation infiltration of psoriatic mouse skin lesions

TABLE 3 activation count of CD3+ T cells at skin lesion site in psoriatic mice

Grouping	CD3+ T (Positive cell number)
		Blank control	1.41±0.37
Model set	13.42±1.95
		Example 3	3.06±0.24*
Comparative example 1	7.38±2.03*△
		Comparative example 2	7.57±2.93*△
Comparative example	12.03±1.76△

As shown in table 3, CD3 marked activated T lymphocytes in the skin, with a positive display of brown. In the blank control group, a small amount of brown positive particles can be seen in the dermis, and the number of positive points in the model group is obviously increased (P is less than 0.05); the example 3 group showed a significant reduction in CD3+ T cells (P <0.05) compared to the model group, as did the control examples 1-2, but not as much as the example 3, and the control group was comparable to the model group.

The results of the above studies show that:

1) the effect of example 3 (with caprylic capric triglyceride, without perilla seed oil) and the effect of comparative example 1 (with caprylic capric triglyceride and without perilla seed oil) and comparative example 2 (with caprylic capric triglyceride and without ascorbic acid ester) are also very significant for the improvement of psoriasis scales, erythema and other symptoms (PASI score), and the effect is different from that of example 3 significantly. The perilla seed oil and the ascorbyl ester have a treatment effect on psoriasis in cooperation with caprylic capric triglyceride, and the synergistic effect is generated by combining the perilla seed oil and the ascorbyl ester with the caprylic capric triglyceride, so that the effect is the best;

2) the detection result of the water content of the psoriasis skin lesion area shows that: example 3 (with caprylic/capric triglyceride, perilla seed oil and ascorbate) and comparative examples 1 (with caprylic/capric triglyceride and without perilla seed oil) and 2 (with caprylic/capric triglyceride and without ascorbate) significantly increased skin moisture content (reflecting skin permeation barrier), but there was no significant difference in the effects between the three, indicating that perilla seed oil and ascorbate had little effect on skin permeation barrier function;

3) the detection results of psoriasis skin lesion area thickness, inflammatory cell infiltration, CD3T cell positive and the like show that: example 3 (with caprylic capric triglyceride, perilla seed oil and ascorbate) significantly improved inflammatory response in the skin lesions of psoriasis. Compared with the comparative example 1 (with caprylic capric triglyceride and without perilla seed oil) and the comparative example 2 (with caprylic capric triglyceride and without ascorbic acid ester), the skin inflammation reaction can be improved, but the effect is not as good as that of the example 3(p is less than 0.05), which shows that the perilla seed oil and the ascorbic acid ester have the effect of inhibiting the inflammation immune reaction in cooperation with the caprylic capric triglyceride, and the synergistic effect can be generated by combining the three.

4) The comparative examples have weak improvement effects on PASI scores, moisture content of skin lesion areas, epidermal thickness, inflammatory cell infiltration, CD3T positive cells and the like, and cannot be used as external preparations for psoriasis;

in conclusion, the product has definite curative effect when being used as an external preparation for psoriasis, and in the raw materials of the product, caprylic capric triglyceride, perilla seed oil and ascorbyl ester are indispensable key raw materials.

Study two: evaluation of improving Permeability skin Barrier function

Evaluation of skin permeation barrier function in psoriasis imiquimod cream mouse model

A mouse model of the imiquimod cream psoriasis is established, the mice are divided into 6 groups such as blank control, a model group, example 3, control example 4, comparative example and the like, 10 mice are fed in each group, and each group is fed in a single cage.

The barrier effect of the skin lesion part on water penetration is examined.

The administration method comprises the following steps: each group was treated with 5 individuals except for the blank control group, 62.5mg of 5% imiquimod cream was applied to the skin for external use, and the groups except for the model group were applied with 50mg of the corresponding test cream (ointment prepared in example 3 or control 3/4 or comparative example) to the corresponding groups after 2 hours of application of the imiquimod cream, and after 1 hour, the skin lesion area was covered with a saturated cotton strip, fixed with a tape, and allowed to intervene for 5 days 1 time a day continuously, and died on day 6.

The barrier effect of the skin lesion part on the penetration of imiquimod is inspected, and the administration method comprises the following steps: each group was prepared by taking 5, giving 50mg of test cream except for the blank control group, applying 62.5mg of 5% imiquimod cream after 1h, 1 time daily, continuously intervening for 5 days, and killing on day 6.

Observation indexes are as follows: performing PASI scoring and epidermis water content and epidermis thickness detection on the skin damage of the mouse; modeling methods and PASI scores, determination of epidermal water content and epidermal thickness were the same as in "study one". The research result shows that:

1) barrier effect of psoriasis skin damage part on water penetration

TABLE 4 psoriasis mice skin lesion PASI score, moisture content and thickness (mean)

As shown in table 4, the psoriatic lesions had a weak barrier to penetration of moisture in the environment, and the model group had significantly increased skin moisture content and lesion thickness, and a higher PASI score. While example 3 blocked water penetration and reduced the PASI score, the skin thickness was closest to the blank, and the improvement of comparative examples 3-4 was less than example 3 but more than the comparative and model groups.

2) Penetration barrier function of psoriasis skin damage part on imiquimod cream

TABLE 5 psoriasis mice skin lesion PASI score, moisture content and thickness

Note: *: compared with the model group, the significant difference (p is less than 0.05); and (delta): compared with example 3, the difference is significant (p < 0.05).

As shown in table 5, the PASI score was significantly increased, skin moisture content was significantly decreased and skin lesion thickness was significantly increased in the model group compared to the blank group, while PASI, skin moisture content and thickness were significantly improved when the imiquimod cream was applied after example 3. The improvement effect of comparative examples 3 to 4 was weaker than that of example 3, but stronger than that of comparative example and model group.

The results of the above two studies show that:

1) for the improvement of psoriasis scales, erythema and other symptoms (PASI score), the effect of example 3 (with hydrogenated lecithin and ceramide, white beeswax and cholesterol) is very obvious, and the effect of control 3 (without hydrogenated lecithin, with ceramide, white beeswax and cholesterol) and control 4 (without ceramide, with hydrogenated lecithin, white beeswax and cholesterol) is also certain, which is obviously different from that of example 3. The combination of the hydrogenated lecithin and the ceramide has a treatment effect on psoriasis, and the combination of the hydrogenated lecithin and the ceramide and the white beeswax and cholesterol has a best effect;

2) the detection result of the water content of the psoriasis skin lesion area shows that: example 3 (with hydrogenated lecithin and ceramide, white beeswax and cholesterol) and control 3 (without hydrogenated lecithin, with ceramide, white beeswax and cholesterol) and control 4 (without ceramide, with hydrogenated lecithin, white beeswax and cholesterol) can obviously block the external moisture and the imiquimod from permeating (reflecting the skin permeation barrier), but the significant difference exists between the control 3 and the control 4 and the example 3, which shows that the hydrogenated lecithin and the ceramide have the function of improving the skin permeation barrier in cooperation with the white beeswax and the cholesterol, and the combination of the four has the synergistic effect and the best effect;

3) the comparative example has weak effects on PASI score, water infiltration resistance of skin lesion areas and imiquimod infiltration, and cannot be used as an external preparation for psoriasis;

in conclusion, the product has definite curative effect as an external preparation for psoriasis, and the raw materials of the product comprise hydrogenated lecithin, ceramide, white beeswax and cholesterol which are indispensable key raw materials.

And (3) research III: functional evaluation research affecting skin' epidermis symbiotic bacteria

Evaluation of efficacy evaluation of skin 'epidermal symbiotic bacteria' affecting Imquimod cream mouse model of psoriasis

Culturing symbiotic bacteria on the skin surface, namely standard strains of staphylococcus aureus ATCC25923, candida albicans ATCC10231, streptococcus pyogenes ATCC19615, staphylococcus epidermidis ATCC12228, propionibacterium ATCC6919 and malassezia ATCC44344, wherein the standard bacteria are all purchased from Bauscere organisms, culturing for 20 hours by adopting nutrient broth, and mixing bacterial liquids of all bacteria in the same volume to prepare a compound bacterial liquid.

A psoriasis mouse model of imiquimod cream is established, animals are divided into 7 groups such as blank control, a model group, an example 3, control examples 1-3, a comparative example and the like, 5 animals are fed in each group in a single cage. Except for the blank control group, 62.5mg of 5% imiquimod cream was applied externally 1 time a day. On the first day, after the imiquimod cream is smeared for 4 hours, 1ml of smeared compound bacterial liquid is taken by a cotton swab. On days 2-6, after 2 hours of application of the imiquimod cream, 60mg of the test cream was applied, once a day, and sacrificed on day 7.

Observation indexes are as follows: before the ointment to be tested is applied on the 2 nd and the sixth day, a cotton swab is used for wiping the surface of the skin lesion to take a microbial sample for gene sequencing (Meige V1V9), and the abundance of various bacteria is analyzed. Performing PASI scoring and detection of epidermis water content and skin damage thickness on the skin damage of the mouse before smearing the paste to be detected on the 2 nd day and the 6 th day; other operations were the same as in "efficacy evaluation study 1".

Three results of the study:

1) abundance analysis of epidermal symbiotic bacteria at skin lesion part of psoriasis mouse

TABLE 6 psoriasis mouse skin lesion symbiotic bacteria abundance

Note: *: compared with the abundance of D2 bacteria in the same group, the abundance of D6 bacteria has significant difference (p <0.05) 2) PASI score, water content and thickness of skin lesions of psoriasis mice.

As shown in table 6, changes in the abundance of bacteria on day six were examined with the abundance of bacteria on day two as a reference. The abundances of staphylococcus aureus, candida and streptococcus are obviously increased after skin damage occurs in the model group and the comparative example group, the abundances of staphylococcus epidermidis, propionibacterium and malassezia are obviously reduced, and the like, while the examples 3 and the comparative examples 1-3 have prebiotic effects, wherein the effects of the examples 3 and the comparative examples 3 are stronger than those of the comparative examples 1-2; no prebiotic effect was shown for the control.

TABLE 7 psoriasis mice skin lesion PASI score, Water content and thickness of skin lesion

As shown in table 7, the PASI scores of the model group and the comparative example group were significantly increased, the skin moisture content was significantly decreased, and the thickness of the skin lesion was significantly increased, compared to the blank group, while the PASI, the skin moisture content, and the thickness of example 3 and comparative examples 1 and 2 were significantly improved, wherein the effect of example 3 was stronger than that of comparative examples 1-2; comparative example 3 and comparative example were poor in effect.

The three results of the study show that:

1) the prebiotics effect is strongest on the paragonial bacteria in the psoriasis skin lesion area in example 3 and comparative example 3 (both containing perilla seed oil, ascorbate and caprylic/capric triglyceride), and secondly, the prebiotics effect is also certain in comparative example 1 (containing no perilla seed oil, ascorbate and caprylic/capric triglyceride) and comparative example 2 (containing no ascorbate, perilla seed oil, ascorbate and caprylic/capric triglyceride), and the prebiotics effect is not in comparative example. The perilla seed oil, the ascorbic acid ester and the combined caprylic capric triglyceride are key raw materials with prebiotics effect, and the prebiotics function of the combination of the three raw materials is optimal.

2) In terms of moisture content in the skin lesion area of psoriasis, the effect of example 3 and comparative examples 1-2 (with hydrogenated lecithin and ceramide, cholesterol, and white beeswax) is very significant, and the effect of comparative example 3 (without hydrogenated lecithin, with ceramide, cholesterol, and white beeswax) in preventing water loss is reduced, indicating that hydrogenated lecithin is very important for the recovery of the permeability barrier.

3) In the aspects of thickness of skin lesion areas of psoriasis, PASI score and the like, the effect of the example 3 and the control examples 1-2 is very obvious, and the effect of the control example 3 (containing perilla seed oil and ascorbic acid ester and no hydrogenated lecithin) is poor. It is indicated that the consideration of the 'epidermal symbiotic bacteria prebiotic' effect and the 'skin permeation barrier' function is the key to ensure the treatment of psoriasis.

In a word, the product has definite curative effect when being used as an external preparation for psoriasis, and the raw materials of the product comprise hydrogenated lecithin, ceramide, perilla seed oil, ascorbic acid ester, cholesterol, white beeswax, caprylic capric triglyceride and the like which are all indispensable key raw materials of the product.

And (4) researching a fourth step: evaluation of melting Point of Mixed fat/oil for skin feeling in use

A series of different melting point formulations (see Table 8) were prepared according to the following table, and the ointment in Table 8 was evaluated for skin feel. The test method for crowd evaluation comprises the following steps: 18-45-year-old volunteers were selected for 30 cases. The test period was six weeks, daily, weekly evaluating an ointment, application method: a proper amount of the skin lipid membrane component is smeared on the back of the hand to experience the skin feel, and the result of crowd evaluation is shown in Table 9.

TABLE 8 raw material ratios and melting point ranges for different greases

The evaluation of skin feel index adopts a grading system. The skin feel after use was specifically graded as:

level 1: after the skin care product is used, the skin has no adverse reaction, is not dried, and is fresh and moist.

And 2, stage: after the skin care product is used, the skin has no adverse reaction, and cannot be dry and wet.

And 3, level: after the skin care product is used, the skin does not have adverse reaction, and the feeling of the skin care product is greasy or very dry.

TABLE 9 evaluation of skin feel after application of different melting Point greases

Evaluation index

Ointment 1

Ointment 2

Ointment 3

Example 3

Example 5

Ointment 7

After-use skin feel

Grade 3

Stage 2

Level 1

Stage 2

Number of people

30 persons

The results of the investigation showed that the melting point of the mixed fat was adjusted to be in the range of 45 to 65 ℃ for the best feeling in use.

In a word, the product has the efficacies of epidermal symbiotic bacteria prebiotics in the psoriasis skin lesion area, improves the 'permeation barrier function', improves the symptoms of pruritus, scale, erythema and the like, reduces the activation of T cells, reduces the recurrence and reduces the psoriasis severity, and is a brand-new external preparation for treating psoriasis. In the formula of the product, caprylic acid capric triglyceride, perilla seed oil and ascorbic acid ester are key raw materials for the product to play a prebiotics function, white beeswax, cholesterol, ceramide, hydrogenated lecithin and the like are key raw materials for playing a role in physical barrier effect, and squalane is an important raw material for adjusting the melting point of mixed oil and improving the skin affinity of a solid oil film. The curative effect of the product for treating psoriasis can be guaranteed as long as the key raw materials are adjusted and prepared within the scope of the right of the application, so that small amounts of other auxiliary components (including spices and other animal and vegetable oil and fat) are added on the basis of the proportion of the key raw materials and the important raw materials, and the invention still belongs to the protection scope of the invention.

The present invention is not limited to the above alternative embodiments, and any other various products can be obtained by anyone in the light of the present invention, and all fall within the protection scope of the present invention. The above-described embodiments should not be construed as limiting the scope of the present invention, and it will be understood by those skilled in the art that modifications may be made to the technical solutions described in the above-described embodiments, or that equivalent substitutions may be made to some or all of the technical features thereof, without departing from the scope of the present invention, and at the same time, such modifications or substitutions may not make the essence of the corresponding technical solutions depart from the scope of the embodiments of the present invention.

Claims

1. A composite composition suitable for psoriasis, wherein the composition is prepared by mixing prebiotic oil, permeation barrier oil and melting point adjusting oil;

the melting point adjusting grease is squalane 2-9.5 parts.

2. The composite composition suitable for psoriasis according to claim 1, wherein the prebiotic oil and fat is prepared from the following raw materials in parts by mass: 45-52 parts of caprylic/capric triglyceride, 1.5-3 parts of perilla seed oil and 5-13 parts of ascorbic acid ester;

3. The composite composition suitable for psoriasis according to claim 1, wherein the permeation barrier oil is prepared from the following raw materials in parts by mass: 9-12 parts of cholesterol, 23.5-31 parts of white beeswax, 0.4-0.9 part of ceramide and 0.5-1.7 part of hydrogenated lecithin;

4. The composite composition suitable for psoriasis according to claim 1 wherein the melting point adjusting oil is squalane 4 to 9 parts;

preferably, the melting point adjusting oil is squalane 6 parts.

5. The combination composition suitable for psoriasis according to any of claims 1 to 4 wherein the ascorbyl esters comprise one or more of ascorbyl palmitate, ascorbyl dipalmitate, ascorbyl methylsilanol pectate, ascorbyl tocopherol maleate, ascorbyl tetraisopalmitate, ascorbyl stearate, tetrahexyldecyl ascorbate, aminopropanol ascorbyl phosphate;

ascorbyl palmitate is preferred.

6. Composite composition suitable for psoriasis according to any of claims 1 to 4, wherein the ceramide is ceramide phosphorylcholine and/or ceramide phosphorylethanolamine.

7. The composite composition suitable for psoriasis according to any of claims 1 to 4, wherein the hydrogenated lecithin is soybean hydrogenated lecithin.

8. A method of preparing a composite composition as claimed in any one of claims 1 to 7, suitable for use in psoriasis, the method comprising:

9. The method for preparing a composite composition suitable for psoriasis according to claim 8 wherein the mixing conditions are:

10. The process for preparing a composite composition suitable for psoriasis according to claim 8 wherein the ingredients are cooled to a temperature of from 40 to 50 ℃ after mixing and stirring.