Use of TPPU and product thereof, and use of TPPU and product thereof

Abstract

The present invention relates to the technical field of ameliorating sensitive skin, and in particular, to use of TPPU in the preparation of a product for ameliorating skin sensitivity, the product, and a method for applying TPPU or the product thereof to ameliorate skin sensitivity. Specifically provided is use of TPPU in the preparation of a product for ameliorating skin sensitivity and / or promoting skin soothing, or a method for applying TPPU to ameliorate skin sensitivity and / or for skin anti-aging. TPPU itself can ameliorate skin sensitivity and / or promote skin soothing and produce a good effect, particularly on skin-sensitive individuals.

Description

Applications of TPPU and its products, and the use of TPPU and its products Technical Field

[0001] This application relates to the technical field of improving skin conditions, and more particularly to the application, products, and use of TPPU in the preparation of products for improving skin sensitivity and / or promoting skin soothing. Background Technology

[0002] TPPU is a compound among soluble epoxide hydrolase inhibitors that inhibits the activity of soluble epoxide hydrolase. Its chemical name is 1-(1-propionylpiperidin-4-yl)-3-(4-(trifluoromethoxy)phenyl)urea, CAS number 1222780-33-7, molecular weight: 359, molecular formula: C 16 H 20 F3N3O3 has the following chemical structural formula (I):

[0003] The aforementioned chemical components are known to have functions and effects focused on treating inflammation or specific pathological conditions of internal organs, nerves, and blood vessels, such as neuropathic pain, mental disorders, and vascular structural / functional disorders. See CN101084216A and CN101505797A for reference.

[0004] In addition to the conventional applications for treating diseases mentioned above, there are reports in the literature that sEH inhibitors can be used to penetrate the skin and reach blood vessels to treat microvascular structural / functional disorders caused by diabetic diseases, such as ulcers. Summary of the Invention

[0005] While the aforementioned literature reports on the treatment of specific inflammations or lesions by sEH inhibitors and discloses methods for skin application of the ingredient, it is known from these documents that 50% DMSO is used as a solvent and penetrant for sEH inhibitors to promote their penetration into the skin and subcutaneous tissue, thereby treating structural / functional lesions of microvessels. However, DMSO itself is not a skin-friendly ingredient, and high concentrations of DMSO may actually increase skin sensitivity, especially when the subject has sensitive skin. Using DMSO as a solvent and penetrant may further increase the subject's skin sensitivity, leading to adverse effects. Therefore, based on the experimental conclusions of the existing literature, the applicant applied TPPU to the skin of the subjects and concluded that TPPU itself can improve skin sensitivity, especially showing good efficacy in those with sensitive skin.

[0006] Therefore, this application provides the use of TPPU in the preparation of products for improving skin sensitivity and / or promoting skin soothing, or a method for using TPPU to improve skin sensitivity and / or promote skin soothing. This method or application can effectively improve skin sensitivity, for example, by effectively preventing, alleviating, or treating skin sensitivity conditions or improving the sensitivity level of sensitive skin. The prevention of skin sensitivity can effectively provide, for example, protection against ultraviolet rays and sun exposure, or prevent, alleviate, or treat skin allergy symptoms such as redness, swelling, pain, and microvascular dilation caused by sunburn, topical products, etc. The relief or treatment of skin sensitivity can effectively alleviate and treat the aforementioned skin allergy symptoms, thereby reducing patient suffering.

[0007] This application found in its research that, compared to the aforementioned literature descriptions, TPPU, when applied to skin cells or tissues, can effectively prevent and alleviate skin allergy symptoms, improve allergic skin, or promote skin soothing. TPPU acts as a sEH inhibitor. Existing research indicates that sEH, as a soluble epoxide hydrolase, can convert fatty acid epoxides produced by P450 enzymes into corresponding diols. sEH inhibitors can inhibit this conversion process, thereby increasing the level of fatty acid epoxides and decreasing the level of diols, such as the increase in 12,13-epoxyoctadecenoic acid. It is currently known that sEH inhibitors can effectively exert anti-inflammatory effects on nerves, organs, and blood vessels. However, TPPU, as one of the sEH inhibitors, can effectively prevent or alleviate skin allergy symptoms, such as skin redness, swelling, skin tissue pain, and microvascular dilation, after being applied to the skin. Experimental confirmation of this can be found in the subsequent examples section.

[0008] The skin sensitivity described in this application includes, but is not limited to, redness, swelling, pain, itching, and telangiectasia of the epidermis, dermis, and subcutaneous tissue. The skin sensitivity described in this application does not refer to skin problems caused by disease or lesions, such as microvascular structural / functional disorders or ulcers. The skin sensitivity described in this application mainly refers to skin allergy symptoms caused by external factors, without any substantial structural or functional lesions occurring in the skin, skin cells, tissues, or organs.

[0009] The products described in this application can be used for pharmaceutical purposes, or for cosmetic or skincare purposes. They can be topical applications or minimally invasive injectable products. Topical applications can be applied directly to the skin surface, while minimally invasive injectable products can be injected subcutaneously via microneedles to better achieve therapeutic effects. The products described in this application are not limited to specific formulations and can be gels, solutions, suspensions, creams, or patches, etc. Any formulation suitable for topical applications or minimally invasive injectable products is acceptable.

[0010] The skin sensitivity described in this application can originate from any of the following: skin care products, cosmetics, topical medications, or sunlight exposure. The skin allergy described in this application is related to individual constitution, primarily to sensitive skin. This sensitivity is not a disease, but rather an allergic symptom caused by changes in skin homeostasis. The allergens mentioned vary depending on individual constitution, and the causes of individual allergies are diverse and not limited to those listed above.

[0011] Sensitive skin differs from skin allergies. It is generally considered a metastable state characterized by reduced tolerance and increased sensitivity to various internal and external factors. The formation mechanism of sensitive skin is complex, and common possible mechanisms include, for example, impaired skin barrier function, increased vascular reactivity, and immune and inflammatory responses.

[0012] Damage to the skin barrier primarily refers to damage to the "brick wall" structure of the skin's surface—composed of stratum corneum cells and the lipids and natural moisturizing factors between them, with a sebum film attached to its surface. This damage leads to an imbalance in the skin's surface microecology, resulting in the loss of moisture and nutrients, and the invasion of external bacteria. Ultimately, the epidermis thins, weakening its resistance to external factors and making it more susceptible to external stimuli. This vicious cycle can easily lead to sensitive skin and even trigger inflammatory reactions.

[0013] In individuals with sensitive skin, TRPV-1 is overexpressed, and the more frequent the expression, the more severe the skin sensitivity. Dynamic optical coherence tomography (OCT) measurements revealed increased vascular reactivity in individuals with sensitive skin. Compared to normal skin, sensitive skin exhibits higher vascular density and more superficial microvessels closer to the epidermis. These increased superficial vessels are more susceptible to external stimuli, leading to excessive vascular response and the release of inflammatory mediators. Furthermore, the vasodilation of individuals with sensitive skin is stronger than that of those with normal skin.

[0014] In addition to increasing vascular reactivity, TRPV-1 can also induce neurogenic inflammation by promoting the secretion of endothelin, which leads to mast cell degranulation. Under external stress, it can cause skin reactions such as erythema and edema, making the skin sensitive.

[0015] The factors that trigger sensitive skin are usually diverse and can be divided into exogenous factors (living environment, daily behavior, physical or chemical stimulation) and endogenous factors (race, genetic inheritance, endocrine, etc.).

[0016] Sensitive skin is currently believed to be a complex process involving the skin barrier, neurovascular system, and immune inflammation. Common clinical sensory characteristics of allergic skin include stinging, redness, itching, and scaling.

[0017] In the product described in this application, the effective dosage of TPPU is not subject to many restrictions, but is preferably 0.001 g / cm³. 2~0.5g / cm 2 It can also be preferably 0.01 g / cm³. 2 ~0.2g / cm 2 or 0.05g / cm 2 ~0.1g / cm 2 .

[0018] In this application, TPPU itself can prevent, alleviate, or treat skin sensitivity, but it is also preferable to mix TPPU with a polar protic solvent for application / use to dissolve the TPPU and better prepare it into an article. The polar protic solvent is preferably one or more of ethylene glycol, propylene glycol, isopropanol, butylene glycol, hexanediol, and water. Propylene glycol or isopropanol is particularly preferred.

[0019] When the TPPU is mixed with a polar protic solvent, the mass percentage of TPPU is preferably 1% to 50%, more preferably 5% to 30%, and even more preferably 5% to 10%. Polar protic solvents differ from other solvents in that they mostly contain trace amounts of H₂. + It is a solvent, distinct from polar aprotic solvents such as DMSO.

[0020] In this application, TPPU can preferably be used in combination with any or more of the following: a polar protic solvent, a penetration enhancer, and an adhesive. Mixing TPPU with a polar protic solvent can promote its micro-dissolution; adding a penetration enhancer allows it to penetrate more effectively through the skin into the subcutaneous tissue, resulting in better therapeutic effects. The penetration enhancer is preferably one or more of squalane, cetyl ester, and sodium hyaluronate. Squalane is particularly preferred.

[0021] The mass ratio of the total mass of the TPPU and the polar protic solvent to the mass of the penetration enhancer is preferably 1:(0.1-50), more preferably 1:(0.2-20), and even more preferably 1:(0.5-10).

[0022] The mass ratio of the total mass of the TPPU and the polar protic solvent to the mass ratio of the penetration enhancer and the adhesion agent is preferably 1:(0.1-50):(0.1-50), more preferably 1:(0.2-20):(0.2-20), and even more preferably 1:(0.5-10):(0.5-10).

[0023] Therefore, the specific solution provided in this application is as follows:

[0024] The application of TPPU or the use of TPPU, wherein the application is the preparation of products that improve skin sensitivity and / or promote skin soothing using TPPU, and the use is the application of TPPU to improve skin sensitivity and / or promote skin soothing.

[0025] Optionally, the skin sensitivity includes any one or more of the following: microvascular dilation, skin tingling, redness, itching, peeling, and dryness.

[0026] Optionally, the skin includes the epidermis, dermis, and / or subcutaneous microvessels and tissues.

[0027] Optionally, the product is a topical application or a minimally invasive skin injection, or the method of use is a topical application or a minimally invasive skin injection.

[0028] Optionally, the topical product includes any one of cosmetics, skin care products, and topical medicines.

[0029] Optionally, the improvement may include any one of prevention, relief, or treatment.

[0030] Optionally, the dosage of TPPU is 0.01 mg / cm³. 2 ~0.5mg / cm 2 The preferred value is 0.05 mg / cm³. 2 ~0.3mg / cm 2 More preferably 0.1 mg / cm³ 2 ~0.2mg / cm 2 .

[0031] Optionally, the article is any one of gel, solution, suspension, cream or patch.

[0032] Optionally, the skin sensitivity may originate from any one of the following: skin care products, cosmetics, topical medications, or light exposure.

[0033] Optionally, TPPU can be applied / used in combination with polar protic solvents.

[0034] Optionally, TPPU can be used / applied in combination with polar protic solvents and penetration enhancers.

[0035] Optionally, TPPU can be used / applied in combination with polar protic solvents, penetration enhancers, and adhesion agents.

[0036] Optionally, the polar protic solvent is selected from one or more of water, ethylene glycol, propylene glycol, isopropanol, butanediol, and hexanediol.

[0037] Optionally, the TPPU accounts for 1% to 50% of the mass, preferably 5% to 30%, and more preferably 5% to 10%.

[0038] Optionally, the penetration enhancer is selected from one or more of squalane, cetyl ester, and sodium hyaluronate.

[0039] Optionally, the total mass ratio of the TPPU and the polar protic solvent to the mass ratio of the penetration enhancer is 1:(0.1-50), preferably 1:(0.2-20), and more preferably 1:(0.5-10).

[0040] Optionally, the adhesive is selected from one or more of glycerin and petrolatum.

[0041] Optionally, the mass ratio of the total mass of the TPPU and the polar protic solvent to the mass ratio of the penetration enhancer and the adhesion agent is 1:(0.1-50):(0.1-50), preferably 1:(0.2-20):(0.2-20), and more preferably 1:(0.5-10):(0.5-10).

[0042] This application also provides an article for improving skin sensitivity and / or promoting skin soothing, said article comprising TPPU.

[0043] Optionally, the improvement of skin sensitivity and / or promotion of skin soothing includes improving any one or more of the following: microvascular dilation, skin irritation, redness, itching, peeling, and dryness.

[0044] Optionally, the skin includes the epidermis, dermis, and / or subcutaneous microvessels and tissues.

[0045] Optionally, the product may be a topical application or a minimally invasive skin injection.

[0046] Optionally, the topical product includes any one of cosmetics, skin care products, and topical medicines.

[0047] Optionally, the improvement may include any one of prevention, relief, or treatment.

[0048] Optionally, the dosage of TPPU is 0.01 mg / cm³. 2 ~0.5mg / cm 2 The preferred value is 0.05 mg / cm³. 2 ~0.3mg / cm 2 More preferably 0.1 mg / cm³ 2 ~0.2mg / cm 2 .

[0049] Optionally, the article is any one of gel, solution, suspension, cream or patch.

[0050] Optionally, the skin sensitivity may originate from any one of the following: skin care products, cosmetics, topical medications, or light exposure.

[0051] Optionally, the article may also contain a polar protic solvent.

[0052] Optionally, the article may also contain a polar protic solvent and a penetration enhancer.

[0053] Optionally, the article may also include a polar protic solvent, a penetration enhancer, and an adhesion agent.

[0054] Optionally, the polar protic solvent is selected from one or more of water, ethylene glycol, propylene glycol, isopropanol, butanediol, and hexanediol.

[0055] Optionally, the TPPU accounts for 1% to 50% of the mass, preferably 5% to 30%, and more preferably 5% to 10%.

[0056] Optionally, the penetration enhancer is selected from one or more of squalane, cetyl ester, and sodium hyaluronate.

[0057] Optionally, the total mass ratio of the TPPU and the polar protic solvent to the mass ratio of the penetration enhancer is 1:(0.1-50), preferably 1:(0.2-20), and more preferably 1:(0.5-10).

[0058] Optionally, the adhesive is selected from one or more of glycerin and petrolatum.

[0059] Optionally, the mass ratio of the total mass of the TPPU and the polar protic solvent to the mass ratio of the penetration enhancer and the adhesion agent is 1:(0.1-50):(0.1-50), preferably 1:(0.2-20):(0.2-20), and more preferably 1:(0.5-10):(0.5-10).

[0060] This application also provides the use of any of the foregoing products, including applying the product to improve skin sensitivity and / or promote skin soothing.

[0061] Optionally, the method of use is external application or minimally invasive skin injection. Attached Figure Description

[0062] Figure 1 is a diagram illustrating the mechanism of action of TPPU on the skin;

[0063] Figures 2 to 12 are before-and-after photos of patients' skin in clinical trials before and after using this product. Detailed Implementation

[0064] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0065] The above technical solution will be described in detail below with reference to specific embodiments.

[0066] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0067] To demonstrate that TPPU has the effect of improving skin allergies, the following implementation methods will be described.

[0068] Mechanism of action explanation

[0069] TPPU was invented and synthesized by Professor Bruce Hammock, a partner in the applicant's company and a member of the National Academy of Sciences. Professor Hammock and his team have dedicated over a decade to in-depth and systematic research on its in vivo efficacy, mechanism of action, and metabolic kinetics using various animal models, including mice, rats, and cynomolgus monkeys. Current research clearly demonstrates that TPPU is a soluble epoxide hydrolase inhibitor that inhibits the activity of soluble epoxide hydrolase. Its mechanism of action is as follows: Eicosapentaenoic acid (EET) and related epoxide fatty acids (EpFA) are endogenous anti-inflammatory compounds, but they can be converted by soluble epoxide hydrolase into the less biologically active dihydroxyeicosapentaenoic acid (DHETs). Therefore, inhibiting the activity of soluble epoxide hydrolase can significantly increase EET levels, thereby reducing the levels of various related inflammatory factors (IL-12, INF-γ, Tbet, IL17, IL23) in the body, thus suppressing inflammation.

[0070] Professor Bruce Hammock's research clearly demonstrates that the levels of inflammatory cytokines such as IL-12, INF-γ, Thbet, IL17, and IL23 in inflammation-inducing mice (inflammation group) were significantly increased compared to the negative control group (saline group). After treatment with this novel small-molecule compound (inflammation-inducing mouse novel-compound treatment group), the levels of these inflammatory cytokines were significantly reduced, demonstrating its highly efficient ability to inhibit the expression of inflammatory cytokines (Figure 1). (IL23, IL-17, IL-12, and INF-γ are all potent inflammatory factors in vivo; overexpression in vivo can cause various inflammatory responses.) Therefore, TPPU can broadly inhibit the in vivo expression levels of Th1 and Th17 inflammatory cytokines, suppressing the overactivation and overrelease of multiple inflammatory factors in skin cells, reducing inflammatory factor levels, and thus alleviating vasodilation, redness, and swelling. These results demonstrate its significant application prospects and market value as a novel cosmetic ingredient in the field of skin soothing (improving skin irritation, itching, redness, and swelling).

[0071] Figure 1 illustrates its mechanism of action. IL-23, IL-17, IL-12, and INF-γ are all potent inflammatory cytokines in vivo. The accumulation of these inflammatory cytokines causes vasodilation and redness, and is one of the main indicators of skin inflammation. Overexpression in vivo can lead to various inflammatory responses and autoimmune diseases. The experimental results shown in the figure below indicate that the levels of inflammatory cytokines such as IL-12, INF-γ, Tbet, IL-17, and IL-23 in the inflammatory model mice (inflammation group) were significantly higher than those in the negative control group (saline group). After TPPU treatment, the levels of these inflammatory cytokines were significantly reduced, demonstrating a highly efficient ability to inhibit the expression of inflammatory cytokines.

[0072] This study found that sEH is a key epoxide hydrolase in the human body, and its levels are undesirable if too high or too low. Existing conventionally used sEH inhibitors can inhibit inflammatory factors, but they also inhibit sEH, and their high sensitivity leads to a low sEH activity, which is detrimental to skin homeostasis. TPPU application to the skin can promote appropriate sEH levels (low in a non-pathological state) in skin tissue and microvessels. The main model used in existing technologies is the double knockout mouse model, which exhibits low sensitivity (IC50) to other sEH inhibitors such as GSK2256294. 50 =27pM) is 150 times that of TPPU (IC 50 =3.7nM), resulting in excessively low sEH levels in the measured results (pathologically low, or even completely absent). Therefore, the TPPU used in this application is more suitable for direct application to the skin and can effectively improve skin sensitivity and / or promote skin soothing. As shown in Figure 1, TPPU can reduce inflammatory cytokines, but it does not adjust them to an excessively low state.

[0073] Example Description

[0074] For the TPPU component, it was formulated into a product with the following component ratios to facilitate subsequent experimental testing. The specific component ratios are shown in the table below, where the ratios are parts by weight. For example, Example 1 contains 0.01 parts by weight of TPPU and 0.99 parts by weight of ethylene glycol, without other components or containing trace impurities. When using, the dosage can be adjusted according to the area of ​​skin sensitivity to application, typically at the dosage of a conventional topical application or a minimally invasive injection, such as (0.01 mg to 0.5 mg) / cm². 2 .

[0075] The specific composition is as follows:

[0076] Table 1

[0077] The formulations for each example in Table 1 were prepared according to the proportions of the ingredients in Table 1 and with reference to conventional preparation methods for topical products, for subsequent experiments. The preparation methods for the above examples can refer to conventional preparation methods for existing topical products, and the TPPU can also be prepared according to existing conventional preparation methods.

[0078] I. Animal Experiments (Phototoxicity Experiment - Sunscreen)

[0079] 1. Skin safety evaluation of TPPU:

[0080] Laboratory animals: Several adult, healthy, undamaged, common-grade albino guinea pigs were selected. Females were non-pregnant and had never given birth. Initial weight range: 298-339g. Provided by Jiashan County Jin Tu Rabbit Industry Professional Cooperative (production).

[0081] Relevant toxicological tests were conducted in accordance with the "Cosmetic Safety Technical Specifications" (2015 edition). The specific procedures are as follows:

[0082] (a) 24 hours before the test, the skin on both sides of the spine on the back of the guinea pig was shaved with a razor close to the skin, taking care to avoid damaging the animal's skin. According to the method of "7 Skin Phototoxicity Test" in "Chapter 6 Toxicological Test Methods" of the "Cosmetic Safety Technical Specifications" (2015 edition), the hair removal area of ​​the guinea pig was divided into hair removal areas a, b, c, d, e, f, g, and h, each area being 2cm × 2cm.

[0083] (b) The animal was fixed in place. 0.2 g of the test substance (Examples 1-60) was evenly applied to the hair-removed areas a and b after moistening with distilled water. The control (corresponding to the adjuvants used in Examples 1-60, i.e., components other than TPPU) was evenly applied to the hair-removed areas c and d after moistening with distilled water. A separate hair-removed area was left untreated (blank). After 30 min of application, the area was irradiated. The hair-removed areas a, c, and the blank were covered with aluminum foil and secured with tape. The hair-removed areas b and d were irradiated with UVA at 365 nm at a dose of 10 J / cm². 2 .

[0084] (c) Remove residual test substance with warm water after irradiation. Observe skin reaction at 1h, 24h, 48h and 72h respectively. Determine the skin reaction score of each animal according to the "Skin Irritation Reaction Score" in "7 Skin Phototoxicity Test" of "Chapter 6 Toxicological Test Methods" of the "Cosmetic Safety Technical Specifications" (2015 edition).

[0085] Results evaluation: No skin reaction was recorded as 0, and a skin reaction was recorded as 1, indicating that the test substance was phototoxic. The aforementioned experiment was used to evaluate the preventive effect of TPPU on UV-induced allergy.

[0086] The experimental results are shown in Table 2. It can be seen that the results of the phototoxicity test of the test substance TPPU on guinea pig skin are: no skin phototoxicity was observed.

[0087] Table 2 - Skin reaction scores of TPPU on guinea pigs

[0088] II. Safety and toxicity experiments

[0089] Regarding the TPPU component, relevant toxicological tests were carried out in accordance with the "Technical Specifications for Cosmetics Safety" (2015 edition). The toxicological characteristics of the raw material are as follows:

[0090] (I). Materials and methods:

[0091] 1. Test substance:

[0092] 1.1 Physical state: Off-white to white solid.

[0093] 1.2 Preparation method: Since the sample is insoluble in water, vegetable oil was used as the solvent. According to the ratio of adding 5 g of the sample to 20 mL of vegetable oil, it was fully mixed to prepare a test solution for gavage exposure.

[0094] 2. Experimental animals and rearing environment:

[0095] 2.1 Experimental animals: SD rats, SPF grade, 10 rats, 5 males and 5 females. The body weight of males was (180 - 194) g, and the body weight of females was (180 - 192) g. They were provided by Zhejiang Vital River Laboratory Animal Technology Co., Ltd., and the production license number is: SCXK(Zhe)2,024 - 0,001.

[0096] 2.2 Reasons for animal selection: The "Technical Specifications for Cosmetics Safety" (2015 edition) stipulates that rats are one of the preferred animals for acute oral toxicity tests.

[0097] 2.3 Rearing environment: The experimental animals were reared in plastic breeding boxes (L466×W314×H200 mm) in a barrier environment, and the cages were changed twice a week. Environmental conditions: Temperature: (20 - 22)°C, relative humidity: (52 - 60)%, 12-hour alternating lighting. The experimental animal use license number is: SYXK(Shanghai)2,023 - 0,046.

[0098] 2.4 Feed and Drinking Water: The animal feed was provided by Shanghai Zhouyu Biotechnology Co., Ltd., with the registration certificate number: Shanghai Feed Certificate (2021) 04027. The animal drinking water met the requirements of the national GB5749 "Hygienic Standards for Drinking Water". During the entire test period, the animals were allowed to freely eat and drink.

[0099] 3 Instruments and Equipment: Electronic scale model: ACS-3A 90401582

[0100] Electronic balance model: BS310S 91006909

[0101] 4 Test Methods:

[0102] 4.1 Preliminary Test: Select 2 SPF-grade SD rats of each sex and conduct oral gavage exposure on the animals at a dose of 5000 mg / kg. The gavage volume was calculated based on 1 mL / 100 g body weight and the animals were gavaged twice within 24 hours with an interval of 6 hours. Observe and record the poisoning signs and death conditions of the test animals. Based on the death conditions of the rats within 48 hours, determine the formal test dose.

[0103] 4.2 Formal Test: According to the results of the preliminary test, no obvious poisoning signs or deaths were observed in the rats of the preliminary test within 48 hours after gavage. Therefore, based on the results of the preliminary test, the test substance was subjected to a limit test at a dose of 5000 mg / kg body weight, that is, 6 more rats (half male and half female) were orally exposed to this dose again. The gavage volume was calculated based on 1 mL / 100 g body weight and the animals were gavaged twice within 24 hours with an interval of 6 hours.

[0104] 5 Clinical Observation and Examination

[0105] 5.1 Observe once at 0.5 hour, 1 hour, 2 hours, and 4 hours after exposure, and regularly observe every day within the following 14 days, and record the individual conditions of each animal in detail, including clinical status, toxicity signs, and death conditions. Weigh the animals once a week before dosing and after exposure. At the end of the experiment, weigh the animals again.

[0106] 5.2 At the end of the experiment, all animals were anesthetized and then dissected. Macroscopic pathological changes were observed with the naked eye and all macroscopic lesions were recorded. If necessary, pathological histological analysis was performed; if no obvious abnormalities were found during macroscopic dissection, pathological histological examination was not necessary.

[0107] 6 Data Processing:

[0108] Judge the size of LD50 based on the number of animal deaths.

[0109] (2). Results:

[0110] 1 After exposure, no other obvious poisoning symptoms or deaths were observed in all animals. See the following table for details.

[0111] 2. Gross anatomical findings:

[0112] At the end of the experiment, all experimental animals underwent gross anatomical examination, and no organs with gross pathological changes were found. Therefore, no further pathological histological examination was performed.

[0113] (III) Conclusion:

[0114] Under the conditions of this experiment, the test substance 1-(1-propyl-4-piperidinyl)-3-[4-(trifluoromethoxy)phenyl]urea (TPPU) had an acute oral median lethal dose (LD50) of greater than 5000 mg / kg body weight in male and female rats, which is practically non-toxic.

[0115] III. Skin Clinical Trials

[0116] We recruited several patients with sensitive skin, such as those experiencing dryness, peeling, redness, and stinging due to allergies caused by the application of topical products. We then used human efficacy studies (pixel area of ​​skin wheals on the arm, hemoglobin content of skin on the inner forearm, lactic acid stinging, and pruritus scores) to evaluate the soothing effects of the following formulation examples.

[0117] The results are shown in the table below. "Reduction in wheal pixel area on the arm" refers to the percentage decrease in wheal pixel area on the arm skin after 1 hour of product use, compared to "no soothing skincare composition." Similarly, "Hemoglobin content on the inner forearm skin" refers to the percentage decrease in wheal pixel area on the inner forearm skin after 5 minutes of product use, compared to "no soothing skincare composition." The reduction in lactic acid stinging and itching scores refers to the percentage decrease in lactic acid stinging and itching scores after 5 minutes of product use, respectively, compared to "no soothing skincare composition." Figures 2-12 are before-and-after photographs of patients' skin before and after application of the examples in Tables 3 and 7 below. The figures are arranged in order: Figure 2 shows before-and-after photographs of the skin before and after application of the user formulation example in Table 3 (TPPU + propylene glycol): squalane = 1:2; Figure 3 shows before-and-after photographs of the skin before and after application of the user formulation example in Table 3 (TPPU + propylene glycol): squalane = 1:5; Figure 4 shows before-and-after photographs of the skin before and after application of the user formulation example in Table 3 (TPPU + propylene glycol): squalane = 1:10; Figure 5 shows before-and-after photographs of the skin before and after application of the user formulation example in Table 3 (TPPU + propylene glycol); Figure 6 shows before-and-after photographs of the skin before and after application of the user formulation example in Table 3 (water); Figure 7 shows before-and-after photographs of the user formulation example in Table 7 (TPPU + propylene glycol): squalane = 1:10. Figure 8 shows before-and-after photos of skin using the user formulation example in Table 7 with TPPU + propylene glycol: isopropanol = 1:5; Figure 9 shows before-and-after photos of skin using the user formulation example in Table 7 with TPPU + propylene glycol: butylene glycol = 1:5; Figure 10 shows before-and-after photos of skin using the user formulation example in Table 7 with TPPU + propylene glycol: squalane = 1:5; Figure 11 shows before-and-after photos of skin using the user formulation example in Table 7 with TPPU + propylene glycol: squalane: petrolatum = 1:5:5; Figure 12 shows before-and-after photos of skin using the user formulation example in Table 7 with TPPU + propylene glycol: synthetic squalane = 1:5.

[0118] Table 3

[0119] Table 4

[0120] Table 5

[0121] Table 6

[0122] Table 7

[0123] Table 8

[0124] Table 9

[0125] Table 10

[0126] As can be seen from the table above, for skin allergy symptoms caused by sensitive skin, after the topical application or subcutaneous minimally invasive injection of the TPPU-containing product of this application, the allergy symptoms (dry skin, peeling, redness, swelling, itching, etc.) are relieved or even cured.

[0127] The embodiments of this application can be formulated into various dosage forms, such as ointments, suspensions, patches, etc., depending on the actual drug application method.

[0128] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. Application of TPPU or use of TPPU, wherein the application is the preparation of products that improve skin sensitivity and / or promote skin soothing using TPPU, and the use is the application of TPPU to improve skin sensitivity and / or promote skin soothing.

2. The use according to claim 1, characterized in that, The improvement of skin sensitivity and / or promotion of skin soothing includes improving any one or more of the following: microvascular dilation, skin tingling, redness, itching, peeling, and dryness.

3. The use according to claim 1, characterized in that, The skin includes the epidermis, dermis, and / or subcutaneous microvessels and tissues.

4. The use according to claim 1, characterized in that, The product is a topical application or a minimally invasive skin injection, or the method of use is a topical application or a minimally invasive skin injection.

5. The use of claim 1, wherein The external application includes any one of cosmetics, skin care products, and topical medicines.

6. The use of claim 1, wherein The improvement includes any one of prevention, relief, or treatment.

7. The use of claim 1, wherein The amount of the TPPU to be applied is 0.01 mg / cm 2 ~ 0.5 mg / cm 2 , preferably 0.05 mg / cm 2 ~ 0.3 mg / cm 2 , more preferably 0.1 mg / cm 2 ~ 0.2 mg / cm 2 .

8. The use of claim 1, wherein, The product is any one of gel, solution, suspension, cream or patch.

9. The use of claim 1, wherein, The skin sensitivity is caused by any one of the following: skin care products, cosmetics, topical medications, or sunlight exposure.

10. The use of claim 1, wherein, TPPU is used in combination with polar protic solvents.

11. The use of claim 1, wherein, TPPU is used in combination with polar protic solvents and penetration enhancers.

12. The use of claim 1, wherein, TPPU is used in combination with polar protic solvents, penetration enhancers, and adhesion agents.

13. Use of any of claims 10, 11 or 12, wherein the compound is ###0002### The polar protic solvent is selected from one or more of water, ethylene glycol, propylene glycol, isopropanol, butanediol, and hexanediol.

14. The use of claim 10, wherein, The TPPU accounts for 1% to 50% of the total mass, preferably 5% to 30%, and more preferably 5% to 10%.

15. The use of any of claims 11 or 12, wherein the compound is ###0002### The penetration enhancer is selected from one or more of squalane, cetyl ester, and sodium hyaluronate.

16. The use of claim 11, wherein, The total mass ratio of the TPPU and the polar protic solvent to the penetration enhancer is 1:(0.1-50), preferably 1:(0.2-20), and more preferably 1:(0.5-10).

17. The use of claim 12, wherein, The adhesive is selected from one or more of glycerin and petrolatum.

18. The use of claim 12, wherein, The total mass ratio of the TPPU and the polar protic solvent to the mass ratio of the penetration enhancer and the adhesion agent is 1:(0.1-50):(0.1-50), preferably 1:(0.2-20):(0.2-20), and more preferably 1:(0.5-10):(0.5-10).

19. An article of manufacture for improving skin sensitivity and / or promoting skin soothing, characterized in that, The product contains TPPU.

20. The article of claim 19, wherein, The improvement of skin sensitivity and / or promotion of skin soothing includes improving any one or more of the following: microvascular dilation, skin tingling, redness, itching, peeling, and dryness.

21. The article of claim 19, wherein The skin includes the epidermis, dermis, and / or subcutaneous microvessels and tissues.

22. The article of claim 19, wherein The product is a topical application or a minimally invasive skin injection.

23. The article of claim 22, wherein, The external application includes any one of cosmetics, skin care products, and topical medicines.

24. The article of claim 19, wherein, The improvement includes any one of prevention, relief, or treatment.

25. The article of claim 19, wherein, The amount of the TPPU to be applied is 0.01 mg / cm 2 ~ 0.5 mg / cm 2 , preferably 0.05 mg / cm 2 ~ 0.3 mg / cm 2 , more preferably 0.1 mg / cm 2 ~ 0.2 mg / cm 2 .

26. The article of claim 19, wherein The product is any one of gel, solution, suspension, cream or patch.

27. The article of claim 19, wherein The skin sensitivity is caused by any one of the following: skin care products, cosmetics, topical medications, or sunlight exposure.

28. The article of claim 19, wherein, The article also contains a polar protic solvent.

29. The article of claim 19, wherein, The product also contains a polar protic solvent and a penetration enhancer.

30. The article of claim 19, wherein, The product also includes a polar protic solvent, a penetration enhancer, and an adhesion agent.

31. The article of any one of claims 28, 29, or 30, wherein, The polar protic solvent is selected from one or more of water, ethylene glycol, propylene glycol, isopropanol, butanediol, and hexanediol.

32. The article of claim 28, wherein The TPPU accounts for 1% to 50% of the total mass, preferably 5% to 30%, and more preferably 5% to 10%.

33. The article of any of claims 29, 30, wherein, The penetration enhancer is selected from one or more of squalane, cetyl ester, and sodium hyaluronate.

34. The article of claim 29, wherein The total mass ratio of the TPPU and the polar protic solvent to the penetration enhancer is 1:(0.1-50), preferably 1:(0.2-20), and more preferably 1:(0.5-10).

35. The article of claim 30, wherein The adhesive is selected from one or more of glycerin and petrolatum.

36. The article of claim 30, wherein The total mass ratio of the TPPU and the polar protic solvent to the mass ratio of the penetration enhancer and the adhesion agent is 1:(0.1-50):(0.1-50), preferably 1:(0.2-20):(0.2-20), and more preferably 1:(0.5-10):(0.5-10).

37. Use of the article of claim 19, comprising applying the article to improve skin sensitivity and / or promote skin soothing.

38. The use of the article of claim 37, wherein, The method of use is external application or minimally invasive skin injection.

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