Use of a topical formulation comprising crocetin or a pharmaceutically acceptable salt thereof

Abstract

The application discloses a use of a topical preparation containing crocetin or a pharmaceutically acceptable salt thereof. The topical preparation developed in the application is used for treating cheilitis simplex actinica caused by chemotherapy, is convenient to use, has few toxic and side effects, and can effectively relieve symptoms such as cheilitis simplex actinica, lip crack, desquamation, erosion and scabbing caused by chemotherapy. The application also expands the application field of crocetin drugs, and finds a new use of crocetin and the salt thereof, which can be prepared into a topical preparation for preventing / treating skin inflammation or skin damage, especially for preventing / treating cheilitis simplex actinica caused by chemotherapy and skin damage caused by oxidative stress, and has better effects. When the crocetin and the salt thereof are used in combination with chlorogenic acid, they have better therapeutic effects. The crocetin and the salt thereof have higher stability when prepared into the topical preparation, are easier to obtain high-purity products, and are more suitable for pharmaceutical development.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, and in particular to the use of topical preparations containing crocin or its pharmaceutically acceptable salts. Background Technology

[0002] Chemotherapy-induced angular cheilitis is a common ailment among cancer patients undergoing chemotherapy. The severe side effects of high-intensity chemotherapy drugs severely damage the body's immune system during treatment. Combined with the release of free radicals and inflammatory factors from chemotherapy drugs, this can lead to severe and persistent angular cheilitis. This type of angular cheilitis differs from ordinary angular cheilitis caused by infection, which can be treated with anti-infective drugs, but these are largely ineffective against chemotherapy-induced angular cheilitis. While some oral anti-inflammatory, antioxidant, hormonal, and immunosuppressant medications can alleviate symptoms to some extent, they do not precisely target the symptoms of angular cheilitis, have low oral bioavailability, significant toxic side effects, high drug dependence, and are inconvenient to use. Currently, there is a lack of effective topical medications specifically targeting chemotherapy-induced angular cheilitis. Developing symptomatic topical formulations that directly target the affected area would avoid the drawbacks of oral formulations, such as high dosage, relatively high toxicity, and poor patient compliance, significantly improving treatment efficacy.

[0003] In summary, there is an urgent need in this field to develop topical formulations that are effective against chemotherapy-induced angular cheilitis. Summary of the Invention

[0004] The purpose of this invention is to provide the use of a topical preparation containing crocin and its medicinal salts.

[0005] Another object of the present invention is to provide a method for preventing and / or treating angular cheilitis.

[0006] Another object of the present invention is to provide a method for preventing and / or treating skin damage caused by oxidative stress.

[0007] To address the aforementioned technical problems, the first aspect of this invention provides the use of a topical preparation containing crocin or its pharmaceutically acceptable salt, for:

[0008] (i) Prevention and / or treatment of angular cheilitis caused by chemotherapy;

[0009] (ii) Prevention and / or treatment of skin damage caused by oxidative stress;

[0010] (iii) To prepare medicines for the prevention and / or treatment of chemotherapy-induced angular cheilitis;

[0011] (iv) To prepare drugs for the prevention and / or treatment of skin aging caused by oxidative stress;

[0012] (v) Improves chapped, dry, peeling, eroded, and / or crusted lips; and / or

[0013] (vi) Improve skin redness, erythema, blisters and / or erosion.

[0014] In some preferred embodiments, the oxidative stress-induced skin cell damage is skin damage caused by hydrogen peroxide.

[0015] In some preferred embodiments, the topical preparation further includes chlorogenic acid or a pharmaceutically acceptable salt thereof.

[0016] In some preferred embodiments, the mass percentage of crocin or its pharmaceutically acceptable salt and chlorogenic acid or its pharmaceutically acceptable salt is (0.8-1.2):(0.8-1.2).

[0017] In some preferred embodiments, the crocin or its pharmaceutically acceptable salt constitutes 0.5-2% of the total mass of the topical preparation, for example, 1.5%.

[0018] In some preferred embodiments, the chlorogenic acid or its pharmaceutically acceptable salt constitutes 0.5-2% of the total mass of the topical preparation, for example, 1.5%.

[0019] In some preferred embodiments, the topical formulation also includes pharmaceutically acceptable excipients.

[0020] In some preferred embodiments, the pharmaceutically acceptable excipients include emulsifiers, humectants, pH adjusters, and water.

[0021] In some preferred embodiments, the emulsifier is selected from at least one of stearic acid and glyceryl monostearate.

[0022] In some preferred embodiments, the moisturizer is selected from at least one of petrolatum and glycerin.

[0023] In some preferred embodiments, the acid-base regulator is sodium hydroxide.

[0024] In some preferred embodiments, the pharmaceutically acceptable excipients include stearic acid, glyceryl monostearate, petrolatum, glycerin, sodium hydroxide, and water.

[0025] In some preferred embodiments, the topical preparation is used to prevent and / or treat skin cell damage caused by oxidative stress and / or to prepare a drug for preventing and / or treating skin cell damage caused by oxidative stress, which may increase the expression level of the antioxidant enzyme superoxide dismutase; decrease the level of malondialdehyde; and / or increase the hydroxyproline content of epidermal cells.

[0026] A second aspect of the present invention provides a method for preventing and / or treating chemotherapy-induced angular cheilitis, the method comprising the steps of:

[0027] Administer a therapeutically effective amount of a topical preparation containing crocin or its medicinal salts to the subject.

[0028] In some preferred embodiments, the application is performed at least once a day for at least one consecutive day.

[0029] A third aspect of the present invention provides a method for preventing and / or treating skin cell damage caused by oxidative stress, the method comprising the steps of:

[0030] Administer a therapeutically effective amount of a topical preparation containing crocin or its medicinal salts to the subject.

[0031] In some preferred embodiments, the oxidative stress-induced skin cell damage is skin cell damage caused by hydrogen peroxide.

[0032] In some preferred embodiments, the application is performed at least once daily for at least one day.

[0033] A fourth aspect of the present invention provides a topical preparation comprising: crocin or a pharmaceutically acceptable salt thereof; and pharmaceutically acceptable excipients.

[0034] In some preferred embodiments, the topical preparation further includes chlorogenic acid or a pharmaceutically acceptable salt thereof.

[0035] Compared with the prior art, the present invention has at least the following advantages:

[0036] (1) This invention has developed a topical preparation for angular cheilitis caused by chemotherapy. It is easy to use, has few toxic side effects, and effectively relieves symptoms such as cracked lips, peeling, erosion, and crusting caused by chemotherapy.

[0037] (2) This invention overcomes the defect that crocin is not suitable for non-injection administration and discovers new uses for crocin acid and its salts. It can be prepared into topical preparations for the prevention / treatment of angular cheilitis caused by radiotherapy or skin damage caused by oxidative stress. Chemotherapy-induced angular cheilitis is more effective. Topical preparations of crocin acid and its salts have higher stability, are easier to obtain high-purity products, and are more suitable for pharmaceutical development.

[0038] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0039] One or more embodiments are illustrated by way of example with reference to the accompanying drawings, and these illustrative descriptions do not constitute a limitation on the embodiments.

[0040] Figure 1 This is a schematic diagram illustrating the protective effect of crocin and its sodium salt against oxidative damage to epidermal cells according to an embodiment of the present invention. **, p < 0.01 vs. model group;

[0041] Figure 2 The high-temperature stability test (60℃ for 8h) of crocin I according to the embodiments of the present invention;

[0042] Figure 3 According to the embodiments of the present invention, the high temperature stability test of crocin (60℃ for 10 days) is performed.

[0043] Figure 4 This is a high-temperature stability test (60℃ for 10 days) of sodium crocin according to an embodiment of the present invention. Detailed Implementation

[0044] Saffron, also known as crocus or saffron crocus, is a traditional Chinese medicine derived from the dried stigmas of the saffron crocus (Crocus sativus), a plant in the Iridaceae family. It possesses properties that promote blood circulation, remove blood stasis, cool the blood, detoxify, relieve depression, and calm the mind. The main active ingredient in saffron is crocin, which accounts for over 10% of the medicinal material. Existing literature reports the therapeutic effects of crocin on cardiovascular diseases. However, due to its low stability and difficulty in penetrating biological membranes, crocin has significantly low bioavailability when administered non-injectable. Since most treatments for skin diseases are administered non-injectable, research on the application of crocin in treating skin diseases is limited. The consensus that crocin has low bioavailability when administered non-injectable has deterred researchers in this field from exploring its application value in diseases requiring topical application or other non-injectable routes of administration (such as skin diseases). During their research, the inventors discovered that crocin has better stability and significantly better bioavailability via non-injection routes. Based on this, the inventors unexpectedly discovered that crocin or its pharmaceutically acceptable salts, when prepared into topical formulations, are remarkably effective in treating chemotherapy-induced angular cheilitis.

[0045] compound

[0046] The scope of compounds in this invention includes crocin, its pharmaceutically acceptable salts, and its prodrugs.

[0047] As used in this invention, the term "croceic acid" has the structure shown in Formula I, with CAS number 27876-94-4.

[0048]

[0049] As used herein, the term "pharmaceutical-grade salt" refers to a salt prepared from a pharmaceutically acceptable, non-toxic alkali (including inorganic and organic bases). Where applicable, pharmaceutically acceptable salts include salts derived from pharmaceutically acceptable inorganic or organic bases formed with crocin. Suitable salts include salts derived from alkali metals such as potassium and sodium, alkaline earth metals such as calcium and magnesium, and ammonium salts, as well as numerous other bases well known in the pharmaceutical field. In a preferred embodiment of the invention, "pharmaceutical-grade salts" are sodium, potassium, ammonium, magnesium, calcium, ferrous, and zinc salts.

[0050] Indications

[0051] This invention relates to the use of crocin and its medicinal salts for:

[0052] (i) Prevention and / or treatment of chemotherapy-induced angular cheilitis; (ii) preparation of medicines for prevention and / or treatment of chemotherapy-induced angular cheilitis; (iii) prevention and / or treatment of skin cell damage caused by oxidative stress; (iv) preparation of medicines for prevention and / or treatment of skin cell damage caused by oxidative stress; (v) improvement of cracked, dry, peeling, erosion and / or crusting of the lips; and / or (vi) improvement of skin redness, erythema, vesicles and / or erosion.

[0053] As used in this invention, the term "angular cheilitis" refers to the symptoms such as cracking, dryness, peeling, erosion, and / or crusting of the lips and corners of the mouth caused by severe bodily damage and inflammatory reactions due to reactive oxygen free radicals, inflammatory factors, and other components produced during chemotherapy in cancer patients. Angular cheilitis can cause pain at the corners of the mouth, affecting functions such as speaking and swallowing. In severe cases, it may be necessary to reduce the patient's chemotherapy dosage, thereby affecting the effectiveness of cancer treatment.

[0054] Topical preparations

[0055] The present invention also relates to topical preparations comprising crocin or a pharmaceutically acceptable salt thereof, including crocin or a pharmaceutically acceptable salt thereof; and pharmaceutically acceptable excipients.

[0056] In a preferred embodiment of the present invention, the topical preparation further includes chlorogenic acid (CAS No. 327-97-9) or its pharmaceutically acceptable salt. The inventors have found that when the topical preparation contains both crocin or its pharmaceutically acceptable salt and chlorogenic acid or its pharmaceutically acceptable salt, a significant synergistic effect can be produced, resulting in better treatment of angular cheilitis caused by chemotherapy.

[0057] Preferably, the mass of crocin or its medicinal salt accounts for 0.5-2% of the total mass of the topical preparation.

[0058] Preferably, the mass of chlorogenic acid or its pharmaceutically acceptable salt accounts for 0.5-2% of the total mass of the topical preparation. Preferably, the ratio of m-crocin or its pharmaceutically acceptable salt to m-chlorogenic acid or its pharmaceutically acceptable salt is (0.8-1.2):(0.8-1.2).

[0059] As used in this invention, the term "topical preparation" refers to a preparation applied to the surface of the skin for the purpose of protecting the skin, treating diseases of various parts of the skin, or treating certain systemic diseases, such as powders, lotions, patches, tinctures, liniments, ointments, gels, pastes, aerosols, etc.

[0060] As used herein, the terms "pharmaceuticalally acceptable excipient" and "pharmaceuticalally acceptable carrier" are used interchangeably to refer to pharmaceutically acceptable materials, compositions, or solvents, such as liquid or solid fillers, diluents, solvents, or encapsulating materials. In one embodiment, each component is "pharmaceutically acceptable," meaning that it is compatible with other components in a pharmaceutical formulation and suitable for contact with human or animal tissues or organs without excessive toxicity, irritation, allergic reactions, immunogenicity, or other problems or complications, with a reasonable benefit / risk ratio. In a preferred embodiment of the present invention, "pharmaceuticalally acceptable excipients" are commonly used excipients in the art for preparing formulations administered topically via application, spraying, or topical application. Examples include petrolatum, paraffin, liquid paraffin, stearic acid or its salts, glyceryl monostearate, lanolin, beeswax, cetyl wax, sodium lauryl sulfate, stearyl alcohol, cetyl alcohol, fatty sorbitol, polysorbate, methylparaben, ethylparaben, glyceryl monostearate, sodium alginate, gelatin, benzyl benzoate, dimethicone, glycerin, and sodium hydroxide. In a preferred embodiment of the present invention, pharmaceutically acceptable excipients include emulsifiers, humectants, pH adjusters, and water. The emulsifier is selected from at least one of stearic acid and glyceryl monostearate. The humectant is selected from at least one of petrolatum and glycerin. The pH adjuster is sodium hydroxide.

[0061] In a more preferred embodiment of the present invention, the topical preparation comprises crocin or a pharmaceutically acceptable salt thereof, stearic acid, glyceryl monostearate, petrolatum, glycerin, sodium hydroxide, and water. As an active ingredient, the content of crocin or a pharmaceutically acceptable salt thereof is not less than 1% of the total mass of the topical preparation. As pharmaceutically acceptable excipients, the components can be rationally combined to form liquids, gels, ointments, etc., and the excipients used to form these preparations and their appropriate proportions are well known to those skilled in the art through conventional techniques. Preferably, in this invention, the topical preparation is prepared as an ointment.

[0062] Treatment

[0063] The present invention also relates to the prevention and / or treatment of chemotherapy-induced angular cheilitis, comprising the steps of administering to a subject a therapeutically effective amount of a topical preparation containing crocin or a pharmaceutically acceptable salt thereof.

[0064] As used herein, the term "prevention" refers to the prevention of the onset, recurrence, or spread of a disease or condition, or one or more symptoms associated with such disease or condition. In one embodiment, such symptoms are known to those skilled in the art to be associated with the disease or condition to be prevented. In a particular embodiment, the term refers to the administration of the compound of the invention, with or without other additional active agents, to a patient at risk of developing the disease or disorder described herein before the onset of symptoms. The term includes the suppression and reduction of symptoms of a particular disease. In a particular embodiment, patients with a family history of a disease are specifically considered as candidates. Furthermore, patients with a history of recurrent symptoms are also potential candidates for prevention. In this regard, the term "prevention" may be used interchangeably with the term "preventive treatment."

[0065] As used herein, the term "treatment" refers to the eradication or improvement of a disease or condition, or one or more symptoms associated with such disease or condition. In one embodiment, such symptoms are known to those skilled in the art to be associated with the disease or condition to be treated. In certain embodiments, the term refers to minimizing the spread or aggravation of a disease or condition by administering one or more preventative or therapeutic agents to a subject suffering from such a disease or condition. In some embodiments, the term refers to the administration of the compound of the present invention, with or without other additional active agents, after the onset of symptoms of a particular disease.

[0066] As used in this invention, the term "subject" is defined herein as including animals, such as mammals, including but not limited to primates (e.g., humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In a particular embodiment, the subject is a human.

[0067] As used herein, the term "therapeuticly effective amount" refers to the amount of a compound sufficient to provide a therapeutic effect in the treatment or control of a disease or disorder, or sufficient to delay or minimize one or more symptoms associated with such disease or disorder. A therapeutically effective amount of a compound refers to the amount of a therapeutic agent that, when used alone or in combination with other therapies, can provide a therapeutic effect in the treatment or control of a disease or disorder. The term "therapeuticly effective amount" may include amounts that improve overall therapy, reduce or avoid symptoms or causes of a disease or disorder, or enhance the therapeutic efficacy of another therapeutic agent. Preferred doses of the active compound used in all situations mentioned in this invention are in the range of about 10 ng / kg to 300 mg / kg, preferably 0.1 to 100 mg / kg / day. The compound is conveniently administered in any suitable unit dosage form, including but not limited to unit dosage forms containing less than 1 mg, 1 mg to 3000 mg, preferably 5 to 500 mg of the active ingredient per unit dosage form.

[0068] As used herein, the term "administration" refers to the means of delivering a drug to a subject via various routes, such as oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include penetration enhancers), buccal, sublingual, and suppository forms, in a single dose or multiple doses. In this invention, the topical formulation is administered transdermally.

[0069] Cosmetic Composition

[0070] The present invention also relates to cosmetic compositions comprising crocin or a salt thereof, and cosmetically acceptable excipients.

[0071] As used herein, the term "cosmetics" refers to daily chemical industrial products that are applied, sprayed, or otherwise similarly dispersed on any part of the human body surface (skin, hair, nails, lips, etc.) for the purposes of cleaning, eliminating unpleasant odors, skin care, beautification, and adornment. Exemplary examples include shampoos, conditioners, lotions, creams, body lotions, lipsticks, lip glosses, lip oils, lip masks, face masks, foundations, facial cleansers, essential oils, perfumes, and toners. In a preferred embodiment of the present invention, cosmetics refer to at least one of lip oils, lip glosses, lip masks, lipsticks, and lip balms. In a preferred embodiment of the present invention, cosmetics refer to at least one of lotions, creams, face masks, body lotions, and toners.

[0072] As used in this invention, the term "cosmetic acceptable excipient" means any chemical ingredient added to protect the product, facilitate storage, or facilitate use, such as thickeners, moisturizers, sunscreens, preservatives, surfactants, antioxidants, colorants, fragrances, chelating agents, and film-forming agents.

[0073] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the present invention is further described below in conjunction with specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight. Unless otherwise specified, the experimental materials and reagents used in the following embodiments are commercially available.

[0074] Unless otherwise specified, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. It should be noted that the terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the exemplary embodiments of this application.

[0075] Example 1: Formulation and preparation method of human ointment

[0076] In this embodiment, ointment formulations A and B containing crocin were prepared.

[0077] Prescription A contains the following ingredients per 100g of ointment:

[0078] Stearic acid: 15g;

[0079] Glyceryl monostearate: 3g;

[0080] Vaseline: 10g;

[0081] Sodium hydroxide: 0.5g;

[0082] Glycerin: 10g;

[0083] Crocin: 1g;

[0084] Water: The remainder is water.

[0085] Stearic acid: 15g; Glyceryl monostearate: 3g; Vaseline: 10g; Sodium hydroxide: 0.5g; Glycerin: 10g; Crocin: 1g. Mix the ingredients and slowly add water while stirring continuously until the mixture reaches 100g. Continue stirring until a uniform paste is formed.

[0086] Prescription B contains the following ingredients per 100g of ointment:

[0087] Stearic acid: 15g;

[0088] Glyceryl monostearate: 3g;

[0089] Vaseline: 10g;

[0090] Sodium hydroxide: 0.5g;

[0091] Glycerin: 10g;

[0092] Crocin: 1g;

[0093] Chlorogenic acid: 1g;

[0094] Water: The remainder is water.

[0095] Stearic acid: 15g; Glyceryl monostearate: 3g; Vaseline: 10g; Sodium hydroxide: 0.5g; Glycerin: 10g; Crocinic acid: 1g; Chlorogenic acid: 1g. Mix the ingredients and slowly add water while stirring continuously. Add water to make up to 100g while continuing to stir until a uniform paste is formed.

[0096] Example 2: Exploration of Animal Modeling Methods for Chemotherapy-Induced Angular Cheilitis

[0097] In this embodiment, the method described in the inventor's literature (Zou Xiaolong et al., Research Progress on Animal Models of Chemotherapy-Induced Oral Mucositis, Prevention and Treatment of Oral Diseases, 2020, Vol. 28, 322-326) was used to construct an animal model of chemotherapy-induced angular cheilitis using a combination of chemotherapy drugs and mechanical injury. Experimental Methods:

[0098] SD rats were selected and, after adaptive culture, were intraperitoneally injected with 5-fluorouracil (5-FU) 40 mg / kg for 3 consecutive days. On the 4th day, under anesthesia, the rats' lips were mechanically injured using an injection needle. The severity of angular cheilitis was determined using the Sonis scoring method, with the following scoring criteria:

[0099] 0 points: Lips are normal, without congestion or erosion;

[0100] 1 point: Red spots appear on the lips, but there is no erosion;

[0101] 2 points: Severe erythema and congestion appear on the lips, forming superficial erosions;

[0102] 3 points: Severe redness and congestion on the lips, forming one or more ulcers, but the area does not exceed 1 / 4 of the entire lip;

[0103] 4 points: The ulcer area is close to 1 / 2 of the lip;

[0104] 5 points: The lips show complete patchy ulcers, and the surface of the lips loses its softness.

[0105] Example 3: The therapeutic effect of a topical preparation on angular cheilitis caused by chemotherapy damage

[0106] In this embodiment, the preparation methods of the test drug and placebo are as follows:

[0107] Test drugs: Weigh 50, 100, and 200 mg of crocin (or sodium crocin) respectively, dissolve them in 2 mL of dimethyl sulfoxide (DMSO), add 10 g of petrolatum, mix well, and prepare low, medium, and high dose ointments for later use.

[0108] Placebo: Add 2 mL of DMSO to 10 g of Vaseline, mix well, and set aside.

[0109] In this embodiment, we selected the animal modeling method described in Example 2 to study the therapeutic effect of the drug composition on angular cheilitis caused by chemotherapy damage. The specific grouping was as follows: 10 animals per group.

[0110] Healthy animal group: Experimental animals were not modeled and were given a placebo without drugs;

[0111] Placebo group: Experimental animals were used to establish the model and were given a placebo that did not contain the drug;

[0112] Crocin low (L), medium (M), and high (H) dose groups: experimental animals were modeled and the drugs containing crocin (50, 100, 200 mg) were administered to the animals topically;

[0113] Low (L), medium (M), and high (H) dose groups of sodium crocin: Experimental animals were modeled and the drug containing sodium crocin (50, 100, 200 mg) was administered to the animals externally.

[0114] The experimental method is briefly described below:

[0115] After the experimental animals underwent adaptive culture, they were randomly selected into groups to begin modeling. Depending on the group, the corresponding drug or placebo was applied topically to the lips after modeling treatment (twice daily). After the mechanical injury model was established on the fourth day, medication was continued until days 6, 8, 10, and 12, when scores were recorded. The results are shown in Table 1.

[0116] Table 1. Therapeutic effect of the drug composition on chemotherapy-induced angular cheilitis (score) (mean ± SD, n = 10)

[0117]

[0118] *p < 0.05 vs. model group, **, p < 0.01 vs. model group

[0119] As shown in Table 1 above, compared with the healthy animal group, the model group animals exhibited more typical symptoms of angular cheilitis, and the symptoms of angular cheilitis in the model group gradually improved after the modeling was terminated. Compared with the model group, both crocin and sodium crocin salt groups showed better therapeutic effects, especially the high-dose group, where the symptoms of angular cheilitis were significantly milder than those in the model group, and most data showed significant differences. The active site of crocin and its sodium salt is its acid radical, which is not related to the metal ions that form the salt. At the same dose, the therapeutic effects are similar, and it can be inferred that mixtures of them in different proportions can still produce similar therapeutic effects, similar to the effects of the same dose of crocin.

[0120] Example 4: The therapeutic effect of sodium crocin and chlorogenic acid compound on chemotherapy-induced angular cheilitis

[0121] During the research and development process, we studied the combined effects of sodium crocin and different monomeric drugs. The results showed that when sodium crocin and chlorogenic acid were used in combination, the therapeutic effect on chemotherapy-induced angular cheilitis was significantly enhanced.

[0122] The placebo used in this embodiment is the same as in Example 3.

[0123] In this embodiment, three drug groups were prepared: a high-dose sodium crocin group (200 mg), a high-dose chlorogenic acid group (200 mg), and a sodium crocin group (100 mg) + chlorogenic acid group (100 mg). The preparation method is the same as that in Example 3.

[0124] The facilities in this embodiment are grouped as follows:

[0125] Healthy animal group: Experimental animals were not modeled and were given a placebo without drugs;

[0126] Placebo group: Experimental animals were used to establish the model and were given a placebo that did not contain the drug;

[0127] High-dose sodium crocin group, high-dose chlorogenic acid group, and high-dose sodium crocin + chlorogenic acid group: experimental animals were modeled and three drugs were used respectively.

[0128] To further investigate the therapeutic effect of the compound drug composition on chemotherapy-induced angular cheilitis, the modeling method in this embodiment, based on Example 2, appropriately increased the degree of mechanical injury to make the angular cheilitis symptoms in the model animals relatively more severe. The drug administration method, scoring method, and standards in this embodiment are the same as in Example 3. The final scores and statistical results of each group of treated experimental animals are as follows:

[0129] Table 2. Therapeutic effect of drug composition on chemotherapy-induced angular cheilitis (score) (mean±SD, n=10)

[0130]

[0131] *p<0.05 vs model group, **, p<0.01 vs model group, #p<0.05 vs crocin H group, ##p<0.01 vs crocin H group, △p<0.05 vs chlorogenic acid H group, △△p<0.01 vs chlorogenic acid H group

[0132] As shown in Table 2 above, when used in combination, the drug substitutes (crocin or chlorogenic acid) with the same concentration were not only significantly more effective than the model group, but also better than the crocin H group and the chlorogenic acid H group in most aspects. This result shows that when they are used in combination, they have a certain synergistic effect and can better combat the inflammation of the corners of the mouth caused by chemotherapy drugs.

[0133] Example 5: Protective effect of the pharmaceutical composition against skin cell damage caused by oxidative stress

[0134] The free radical theory of aging posits that the accumulation of free radicals leading to skin cell damage is a major cause of skin inflammation and aging. Ultraviolet radiation, high-energy ionizing radiation used in radiotherapy, and chemotherapy drugs can all generate large amounts of free radicals (which is a key reason why the latter primarily kills cancer cells and damages healthy cells), thereby causing skin inflammation, cell damage, and apoptosis. The dermis, mainly composed of fibroblasts, plays a crucial role in skin inflammation and aging.

[0135] In this embodiment, the inventors followed the method described in the reference (Du Xianhua et al., H2O2-induced oxidative stress damage in cultured skin fibroblasts, Chinese Journal of Gerontology, 2011, Vol. 31, 644-646), selecting human dermal fibroblasts, and using H2O2 to induce oxidative stress damage in the cells to determine the protective effect of the pharmaceutical composition of the present invention against oxidative stress damage. Experimental methods:

[0136] Human dermal fibroblasts were cultured using the method described in the references. After adhering and growing, they were grouped as follows:

[0137] Normal cell group: Normally cultured cells, without H2O2 to induce damage, were treated with the same volume of DMSO solvent for the same time;

[0138] H2O2 treatment group: Cells were treated with 100 μM H2O2 to induce damage (4 h), and the same volume of DMSO solvent was added for the same duration.

[0139] Low (L), medium (M), and high (H) dose groups of crocin: Cells were induced to be damaged by adding 100 μM H2O2 (4 h), and crocin DMSO solution of different concentrations was added at the same time to make the final concentration of crocin in the culture medium 50, 100, and 200 μg / mL, respectively.

[0140] Low (L), medium (M), and high (H) dose groups of sodium crocin: Cells were induced to be damaged by adding 100 μM H2O2 (4 h), and at the same time, different concentrations of crocin DMSO solution were added to make the final concentration of crocin in the culture medium 50, 100, and 200 μg / mL, respectively.

[0141] After the above treatment was completed, cell viability was measured using the MTT assay, and the data were statistically analyzed. The results are as follows: Figure 1 As shown.

[0142] The results showed that crocin and its sodium salt had good effects in combating H2O2-induced oxidative damage. The activity of epidermal cells treated with these substances was significantly increased, and a good dose-response relationship was observed. We speculate that the mechanism of their activity may be related to their antioxidant and anti-apoptotic effects.

[0143] Following the method described in the reference, another batch of cells with different groupings was prepared using the same method. The activities and contents of superoxide dismutase (SOD), malondialdehyde (MDA), and hydroxyproline (HP, a characteristic amino acid of collagen) in the cell culture medium were then determined. The results are shown in Table 3 below.

[0144] Table 3. Effects of the drug composition on H2O2-induced cell damage (mean ± SD, n = 6)

[0145]

[0146] *p < 0.05 vs. model group, **, p < 0.01 vs. model group

[0147] As shown in Table 3 above, crocin and its sodium salt can significantly increase the expression of the antioxidant enzyme SOD, reduce the production of the oxidation product MDA, and increase the content of HP in epidermal cells, with significant differences at medium and high doses.

[0148] Example 6: Comparative Study on the High-Temperature Stability of Crocin and Crocinoside

[0149] Crocin is rare in plants and can be prepared by chemical or biological hydrolysis of crocin (as shown in Formula II below). It has poor water solubility, but can be improved by forming common sodium and potassium salts.

[0150]

[0151] In this embodiment, the inventors will compare the stability of crocin I, crocin acid, and sodium crocinate. The test will be conducted in accordance with the methods for testing high-temperature effects as described in the Chinese Pharmacopoeia (2020 edition, Part IV), and the test will be terminated at appropriate times based on the product's stability.

[0152] Experiments revealed that crocin has very poor stability, undergoing significant degradation within just 4 hours at 60°C. In contrast, crocin acid and its sodium salt exhibit excellent stability, showing only minor degradation even after 10 days at 60°C (see details in [link to results]). Figure 2-4 ).

[0153] The results of this embodiment demonstrate that, compared to crocin, crocinic acid and its sodium salt exhibit significantly higher stability, making them highly suitable for drug development and meeting the "quality controllable" requirements of pharmaceuticals. In contrast, crocin, due to its extremely poor stability, may degrade during its shelf life, rendering it unsuitable as a pharmaceutical raw material. Numerous types of crocin exist in plants (such as I, II, and III), which share similar structures and exhibit strong water solubility, making their separation and purification extremely difficult and hindering their development into pharmaceutical raw materials. Using the crocinic acid fraction, its primary active unit, can significantly reduce purification difficulty and increase product purity.

[0154] In summary, the inventors' research has found that crocin and its sodium salt, when used alone or in pharmaceutical compositions, have significant effects against angular cheilitis caused by radiotherapy and chemotherapy, and dermatitis caused by sun exposure. Furthermore, its high stability makes it suitable for use as a pharmaceutical raw material. Simultaneously, due to crocin's poor water solubility and good lipid solubility, when used as a topical medication or daily chemical product, it is not easily dissolved by water, saliva, sweat, or other bodily fluids. This reduces the difficulty of cleaning the pigment as it flows onto the affected area, making it more convenient to use and thus more suitable for use as a topical preparation.

[0155] Those skilled in the art will understand that the above embodiments are specific examples of implementing the present invention, and in practical applications, various changes in form and detail may be made without departing from the spirit and scope of the present invention.

Claims

1. The use of topical preparations containing crocin and its pharmaceutically acceptable salts and chlorogenic acid or its pharmaceutically acceptable salts for the preparation of drugs for the prevention and / or treatment of chemotherapy-induced angular cheilitis; The crocin and its medicinal salts have a mass percentage content of 0.5-2%; The chlorogenic acid and its pharmaceutically usable salts have a mass percentage content of 0.5-2%.

2. The use according to claim 1, characterized in that, In the topical preparation, the mass percentage of crocin or its pharmaceutically acceptable salt and chlorogenic acid or its pharmaceutically acceptable salt is (0.8-1.2):(0.8-1.2).

3. The use according to claim 1, characterized in that, The topical preparation also includes pharmaceutically acceptable excipients.

4. The use according to claim 3, characterized in that, Pharmaceutically acceptable excipients include emulsifiers, humectants, pH adjusters, and water.

5. The use according to claim 4, characterized in that, The emulsifier is selected from at least one of stearic acid and glyceryl monostearate.

6. The use according to claim 4, characterized in that, The moisturizer is selected from at least one of petrolatum and glycerin.

7. The use according to claim 4, characterized in that, The acid-base regulator is sodium hydroxide.

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