Compounds and methods for treating and / or preventing skin cancer or precancerous lesions thereof

Abstract

The invention relates to 17 beta-dammarin B or pharmaceutically acceptable salts, racemes, stereoisomers, enantiomers, diastereoisomers, tautomers, metabolites, solvates and prodrugs thereof and a method for treating and / or preventing skin cancer or precancerous lesions thereof. The 17 beta-dammarin B is prepared from 17 beta-dammarin B or pharmaceutically acceptable salts, racemes, stereoisomers, enantiomers, diastereoisomers, tautomers, metabolites, solvates and prodrugs thereof. The medicine can permeate through the skin and can directly act on skin lesion parts, the curative effect is improved, and meanwhile high safety is achieved.

Description

Compounds and methods for treating and / or preventing skin cancer or its precancerous lesions

[0001] This application claims priority to and benefits of patent application 202311331579.5 filed with the State Intellectual Property Office of China on October 13, 2023, and incorporates the entire text of it herein by reference. Technical Field

[0002] The present application belongs to the field of medical technology, and specifically relates to compounds and methods for treating and / or preventing skin cancer or its precancerous lesions. Background Art

[0003] Reports indicate that 17β-xanthoside B can target beclin 1, inhibit the formation of LC3-associated phagosomes, and improve the development of experimental autoimmune encephalomyelitis (EAE). Patent No. CN 104736157 A records that 17β-xanthoside B can induce cell cycle arrest and apoptosis in human hepatocellular carcinoma HepG2 cells.

[0004] Skin cancer is the most common malignant tumor in humans worldwide. It is estimated that about 32% of cancers diagnosed each year are skin cancers, which are divided into melanoma skin cancer and non-melanoma skin cancer. The latter is the most common and includes basal cell carcinoma and squamous cell carcinoma of the skin.

[0005] Surgical resection and radiotherapy are traditional treatments for localized non-melanoma skin cancer, but the recurrence rate of surgery or radiotherapy alone is approximately 30%, and some patients are not suitable for surgery or radiotherapy due to disfigurement or inability to achieve radical resection due to anatomical location limitations, or due to various other forms of skin adverse reactions. Most anticancer drugs currently used in clinical practice are administered orally or by injection, and are not suitable for preparation as topical preparations that act directly on skin lesions and cannot be used to treat skin cancer. Topical medications known for the treatment of skin cancer, such as 5-fluorouracil and imiquimod, have limited therapeutic effects. Drugs suitable for the treatment of skin cancer or its precancerous lesions remain to be developed.

[0006] Summary of the Invention

[0007] In one aspect, the present application relates at least in part to a compound, which is 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof, which is topically administered for the treatment and / or prevention of skin cancer or precancerous lesions thereof.

[0008] In one aspect, the present application relates, at least in part, to methods for treating and / or preventing skin cancer or its precancerous lesions, comprising administering 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof.

[0009] In one aspect, the present application relates, at least in part, to a method for treating and / or preventing skin cancer or a precancerous lesion thereof, comprising administering to a subject in need thereof a therapeutically effective amount of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof.

[0010] In one aspect, the skin cancer comprises melanoma and / or non-melanoma skin cancer.

[0011] In one aspect, the non-melanoma skin cancer is basal cell carcinoma and / or squamous cell carcinoma of the skin.

[0012] In one aspect, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof is administered to the affected area of ​​a subject at a dose of about 0.0003 mg to about 100 mg; about 0.008 mg to about 90 mg; about 0.1 mg to about 80 mg; about 0.1 mg to about 70 mg; about 0.1 mg to about 60 mg; about 0.1 mg to about 50 mg; about 0.1 mg to about 40 mg; about 0.1 mg to about 30 mg; about 0.1 mg to about 20 mg; about 0.1 mg to about 15 mg; about 0.1 mg to about 10 mg; about 0.1 mg to about 8 mg; about 0.2 mg to about 5 mg; about 0.5 mg to about 2.5 mg.

[0013] In one aspect, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof is administered to the affected area of ​​a subject at a dose of about 0.02 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 15 ... .9 mg, about 1 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.6 mg, about 1.7 mg, about 1.8 mg, about 1.9 mg, about 2.0 mg, about 2.1 mg, about 2.2 mg, about 2.3 mg, about 2.4 mg, about 2.5 mg, about 2.6 mg, about 2.7 mg, about 2.8 mg, about 2.9 mg, about 3 mg, about 4 mg, or about 5 mg.

[0014] In one aspect, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof is administered to the affected area of ​​a subject at a dose of about 0.0003 mg / cm 2 to about 10 mg / cm 2 ; preferably about 0.001 mg / cm 2 to about 1 mg / cm 2 ; preferably about 0.002 mg / cm 2 to about 0.5 mg / cm 2 ; preferably about 0.004 mg / cm 2 to about 0.25 mg / cm 2 ; preferably about 0.008 mg / cm 2 to about 0.2 mg / cm 2 ; preferably about 0.01 mg / cm 2 to about 0.18 mg / cm 2 ; preferably about 0.016 mg / cm 2 to about 0.16 mg / cm 2 ; preferably about 0.02 mg / cm 2 to about 0.15 mg / cm 2 ; preferably about 0.025mg / cm 2 to about 0.14 mg / cm 2 ; preferably about 0.027mg / cm 2 to about 0.13 mg / cm 2 ; preferably about 0.03mg / cm 2 to about 0.125 mg / cm 2 ; preferably about 0.035mg / cm 2 to about 0.12 mg / cm 2 ; preferably about 0.04 mg / cm 2 to about 0.1 mg / cm 2 .

[0015] In one aspect, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof is administered to the affected area of ​​a subject at a dose of about 0.001 mg / cm 2 , about 0.002mg / cm 2 , about 0.003mg / cm 2 , about 0.004mg / cm 2 , about 0.005mg / cm 2, about 0.006mg / cm 2 , about 0.007mg / cm 2 , about 0.008mg / cm 2 , about 0.009mg / cm 2 , about 0.01mg / cm 2 , about 0.015mg / cm 2 , about 0.02mg / cm 2 , about 0.025mg / cm 2 , about 0.03mg / cm 2 , about 0.035mg / cm 2 , about 0.04mg / cm 2 , about 0.045mg / cm 2 , about 0.05mg / cm 2 , about 0.055mg / cm 2 , about 0.06mg / cm 2 , about 0.065mg / cm 2 , about 0.07mg / cm 2 , about 0.075mg / cm 2 , about 0.08mg / cm 2 , about 0.085mg / cm 2 , about 0.09mg / cm 2 , about 0.095mg / cm 2 , about 0.1mg / cm 2 , about 0.15mg / cm 2 , about 0.2mg / cm 2 , about 0.25mg / cm 2 , about 0.3mg / cm 2 , about 0.35mg / cm 2 , about 0.4mg / cm 2 , about 0.45mg / cm 2 , about 0.5mg / cm 2 , about 0.55mg / cm 2 , about 0.6mg / cm 2 , or about 0.65 mg / cm 2 .

[0016] In one aspect, the subject has an affected area of ​​about 0.01 cm 2 to about 300cm 2 ; Preferably, the affected area of ​​the subject is about 1 cm 2 to about 200cm 2 , about 1cm 2 to about 100cm 2 , about 1cm2 to about 75cm 2 , about 1cm 2 to about 50cm 2 , about 1cm 2 to about 25cm 2 ; about 10cm 2 to about 200cm 2 , about 10cm 2 to about 100cm 2 , about 10cm 2 to about 75cm 2 , about 10cm 2 to about 50cm 2 , about 10cm 2 to about 25cm 2 ; About 25cm 2 to about 200cm 2 , about 25cm 2 to about 100cm 2 , about 25cm 2 to about 75cm 2 ; About 25cm 2 to about 90cm 2 , about 25cm 2 to about 80cm 2 , or about 25cm 2 to about 70cm 2 , about 25cm 2 to about 60cm 2 , about 25cm 2 to about 50cm 2 , about 25cm 2 to about 40cm 2 , or about 25cm 2 to about 30cm 2 Preferably, the affected area of ​​the subject is about 0.2 cm 2 , about 0.5cm 2 , about 1cm 2 , about 2cm 2 , about 3cm 2 , about 4cm 2 , about 5cm 2 , about 6cm 2 , about 7cm 2 , about 8cm 2 , about 9cm 2 , about 10cm 2 , about 11cm 2 , about 12cm 2 , about 13cm 2 , about 14cm 2 , about 15cm2 , about 16cm 2 , about 17cm 2 , about 18cm 2 , about 19cm 2 , about 20cm 2 , about 25cm 2 , about 30cm 2 , about 35cm 2 , about 40cm 2 , about 45cm 2 , about 50cm 2 、approximately 55cm 2 , about 60cm 2 , about 65cm 2 , about 70cm 2 , about 75cm 2 , about 80cm 2 、approximately 85cm 2 , about 90cm 2 , about 95cm 2 , or about 100cm 2 .

[0017] In one aspect, the subject's affected area is skin.

[0018] In one aspect, the subject's affected area is located at one or more locations independently selected from the group consisting of: scalp, forehead, forearm, face, nose, ear, eyelid, lip, neck, arm, hand, torso, leg, and foot.

[0019] In one aspect, the subject has more than one affected area. In one aspect, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof is administered to the patient topically.

[0020] In one aspect, the 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug is prepared into a pharmaceutical composition suitable for external use.

[0021] In one aspect, based on the total weight of the pharmaceutical composition, the content of 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is about 0.025-4 weight%, preferably about 0.05-3 weight%, preferably about 0.2-0.8 weight%, preferably about 0.4-0.8 weight%, preferably about 0.075 weight%, about 0.1 weight%, about 0.125 weight%, about 0.15 weight%, about 0.175 weight%, about 0.2 weight%, about 0.225 weight%, about 0.25 weight%, about 0.275 weight%, about 0.3 weight%, about 0.325 weight%, about 0.35 weight%, about 0.37 5 wt%, about 0.4 wt%, about 0.425 wt%, about 0.45 wt%, about 0.475 wt%, about 0.5 wt%, about 0.525 wt%, about 0.55 wt%, about 0.575 wt%, about 0.6 wt%, about 0.625 wt%, about 0.65 wt%, about 0.675 wt%, about 0.7 wt%, about 0.725 wt%, about 0.75 wt%, about 0.775 wt%, about 0.8 wt%, about 0.825 wt%, about 0.85 wt%, about 0.875 wt%, about 0.9 wt%, about 0.925 wt%, about 0.95 wt%, about 0.975 wt%, about 1 wt%, about 1.25 wt%, about 1.5 wt%, about 1.75 wt%, about 2.0 wt%, about 2.25 wt%, about 2.5 wt%, about 2.75 wt%.

[0022] In one aspect, the pharmaceutical composition is a gel.

[0023] In one aspect, the pharmaceutically acceptable excipients in the gel may optionally include a gel matrix. The gel matrix is ​​at least one selected from sodium carboxymethylcellulose, carbomer, gelatin, and alginate. In one aspect, the gel matrix is ​​present in an amount of about 0.5-5% by weight, preferably about 1-4% by weight, preferably about 1-3% by weight, more preferably about 1.5% by weight, about 2% by weight, about 2.5% by weight, or about 3% by weight, based on the total weight of the pharmaceutical composition.

[0024] In one aspect, the pharmaceutically acceptable excipient in the gel may optionally include a humectant. The humectant is at least one selected from alcoholic humectants; preferably, the humectant is glycerol. In one aspect, the humectant is present in an amount of about 5-35% by weight, preferably about 10-30% by weight, more preferably about 15-25% by weight, and more preferably about 15%, about 20%, about 25%, or about 30% by weight, based on the total weight of the pharmaceutical composition.

[0025] In one aspect, the pharmaceutically acceptable excipient in the gel may optionally include a solvent; preferably, the solvent is at least one of ethanol, propylene glycol, and benzyl alcohol, and the ethanol is 95% ethanol or anhydrous ethanol. In one aspect, the content of the solvent is about 20-75% by weight, preferably about 25-70% by weight, and more preferably about 30% by weight, about 35% by weight, about 40% by weight, about 45% by weight, about 50% by weight, about 55% by weight, about 60% by weight, or about 65% by weight, based on the total weight of the pharmaceutical composition.

[0026] In one aspect, the pharmaceutically acceptable excipient in the gel may optionally include water, preferably purified water, distilled water or deionized water. In one aspect, the water content is about 20-75% by weight, preferably about 25-70% by weight, more preferably about 30% by weight, about 35% by weight, about 40% by weight, about 45% by weight, about 50% by weight, about 55% by weight, about 60% by weight, or about 65% by weight based on the total weight of the pharmaceutical composition.

[0027] On the one hand, the pharmaceutically acceptable excipients in the gel may optionally include a penetration enhancer. On the one hand, the penetration enhancer is at least one selected from azone, borneol, Tween 80, sodium lauryl sulfate, and poloxamer, and more preferably the penetration enhancer is azone or borneol. On the one hand, based on the total weight of the pharmaceutical composition, the content of the penetration enhancer is about 0-5 weight %; preferably about 0.1-4 weight %; more preferably about 0.25 weight %; about 0.5 weight %; about 0.75 weight %; about 1 weight %; about 1.25 weight %; about 1.5 weight %; about 1.75 weight %; about 2 weight %; about 2.25 weight %; about 2.5 weight %; about 2.75 weight %; about 3 weight %; about 3.25 weight %; about 3.5 weight %; about 3.75 weight %. On the one hand, the pharmaceutically acceptable excipients may optionally include a preservative. The preservative is a paraben preservative and / or butylated hydroxytoluene; preferably, the paraben preservative is methylparaben and / or ethylparaben. In one aspect, based on the total weight of the pharmaceutical composition, the preservative is present in an amount of about 0-4% by weight; preferably about 0.1-3% by weight; more preferably about 0.25% by weight; about 0.5% by weight; about 0.75% by weight; about 1% by weight; about 1.25% by weight; about 1.5% by weight; about 1.75% by weight; about 2% by weight; about 2.25% by weight; about 2.5% by weight; about 2.75% by weight.

[0028] In one aspect, the pharmaceutical composition comprises:

[0029] About 0.2-0.8% by weight of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof; and / or

[0030] about 1.5% by weight of a gel matrix; and / or

[0031] about 40% by weight of a solvent; and / or

[0032] About 0.5% by weight of a penetration enhancer; and / or

[0033] about 20% by weight of a humectant;

[0034] and the balance of water.

[0035] In one aspect, the pharmaceutical composition is a topical composition comprising 17β-xanthoside B free base, wherein the 17β-xanthoside B free base is greater than about 90% as determined by HPLC; preferably greater than about 90.5%, greater than about 91%, greater than about 91.5%, greater than about 92%, greater than about 92.5%, greater than about 93%, greater than about 93.5%, greater than about 94%, greater than about 94.5%, greater than about 95%, greater than about 95.5%, greater than about 96%, greater than about 96.5%, greater than about 97%, greater than about 97.5%, greater than about 98%, greater than about 98.5%, greater than about 99%, greater than about 99.5%, greater than about 99.6%, greater than about 99.7%, greater than about 99.8% or greater than about 99.9%.

[0036] In one aspect, dosage forms of topical compositions include powders, sprays, ointments, emulsions, pastes, creams, lotions, gels, solutions, patches, and inhalants.

[0037] In one aspect, the dosage form of the topical composition comprises a gel.

[0038] In one aspect, the amount of the pharmaceutical composition applied to the affected skin area is about 0.001 g / cm 2 Skin surface area: approximately 0.5 g / cm 2 The skin surface area is preferably within the range of about 0.0015 g / cm 2 Skin surface area: approximately 0.2 g / cm 2 Skin surface area; preferably about 0.0015 g / cm 2 Skin surface area: approximately 0.15 g / cm 2 Skin surface area; preferably about 0.002 g / cm 2 Skin surface area: approximately 0.125 g / cm 2 Skin surface area; preferably about 0.003 g / cm 2 Skin surface area: approximately 0.1 g / cm 2 Skin surface area; preferably about 0.004 g / cm 2 Skin surface area: approximately 0.005 g / cm2 Skin surface area: approximately 0.006 g / cm 2 Skin surface area: approximately 0.007 g / cm 2 Skin surface area: approximately 0.008 g / cm 2 Skin surface area: approximately 0.009 g / cm 2 Skin surface area: approximately 0.01 g / cm 2 Skin surface area: approximately 0.0125 g / cm 2 Skin surface area: approximately 0.015 g / cm 2 Skin surface area: approximately 0.0175 g / cm 2 Skin surface area: approximately 0.02 g / cm 2 Skin surface area: approximately 0.03 g / cm 2 Skin surface area: approximately 0.04 g / cm 2 Skin surface area: approximately 0.05 g / cm 2 Skin surface area: approximately 0.06 g / cm 2 Skin surface area: approximately 0.07 g / cm 2 Skin surface area: approximately 0.08 g / cm 2 Skin surface area: approximately 0.09 g / cm 2 Skin surface area.

[0039] In one aspect, 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is administered every day, every other day, every two days, every three days, every four days, every five days, every six days, or every seven days. Typically, the administration day is one, two, three, or four times.

[0040] In one aspect, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof is administered once a week, once every six days, once every five days, once every four days, once every three days, once every two days, once a day, twice a day, three times a day, or four times a day; preferably once every two days or once a day.

[0041] In one aspect, the administration period of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof can be 1 to 100 days, preferably 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 days.

[0042] In one aspect, the frequency of administration of 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug can be once or twice a day, continued for more than one day per week, and then stopped for the rest of the week. In one aspect, it is administered every other day, once or twice a day. In one aspect, it is administered every three days, every four days, every five days, every six days, or every seven days, once or twice a day. In one aspect, it is administered every three days, every four days, every five days, every six days, or every seven days, for two consecutive days, once or twice a day. In one aspect, it is administered every four days, every five days, every six days, or every seven days, for three consecutive days, once or twice a day. In one aspect, it is administered every five days, every six days, or every seven days, for four consecutive days, once or twice a day. In one aspect, it is administered until the skin cancer or its precancerous lesions are completely cured. In one aspect, it is administered topically.

[0043] In one aspect, the drug is maintained in contact with the skin for about 0.5 to about 24 hours per administration, optionally about 1 hour, about 1.5 hours, about 2 hours, about 2.5 hours, about 3 hours, about 3.5 hours, about 4 hours, about 4.5 hours, about 5 hours, about 5.5 hours, about 6 hours, about 6.5 hours, about 7 hours, about 7.5 hours, about 8 hours, about 8.5 hours, about 9 hours, about 9.5 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, or about 24 hours.

[0044] In one aspect, administration of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof reduces the number and / or severity of local skin reactions or other adverse side effects in a subject compared to other treatments for skin cancer or its precursor lesions.

[0045] In one aspect, administration of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof reduces the number of subjects experiencing local skin reactions or other adverse side effects compared to other treatments for skin cancer or its precursor lesions.

[0046] In one aspect, the local skin reaction is selected from the group consisting of vesicle formation, pustule formation, erosion, ulceration, redness, swelling, flaking, scaling, lumps, dryness, pus, and blisters.

[0047] In one aspect, the other side effect is selected from the group consisting of application site pain, application site itching, application site irritation, application site swelling, application site burning, application site infection, peri-ocular edema, nasopharyngitis, chills, sore throat, ptosis, puffy eyes, hypopigmentation, hyperpigmentation, and headache.

[0048] In one aspect, the present application relates at least in part to 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof, which is used for (e.g., topically for) treating and / or preventing skin cancer or its precancerous lesions. In some aspects, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof is used as described herein at the dosage, dosing schedule, and / or in one or more diseased areas in a subject in need thereof.

[0049] In one aspect, the present application relates at least in part to the use of 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug in the treatment and / or prevention of skin cancer or its precancerous lesions (e.g., topical use). In some aspects, 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is used as described herein at the dosage, administration schedule, and / or in one or more diseased areas in a subject in need thereof.

[0050] In one aspect, the present application relates at least in part to the use of 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug in the preparation of a medicament for treating and / or preventing skin cancer or its precancerous lesions. In some aspects, 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is used as described herein at the dosage, administration schedule, and / or in one or more diseased areas in a subject in need thereof.

[0051] In one aspect, the present application relates, at least in part, to a skin cancer or precancerous lesion therapeutic agent comprising 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof. In some aspects, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof is used as described herein at the dosage, dosing schedule, and / or in one or more diseased areas in a subject in need thereof.

[0052] As used herein, the term "skin cancer" is used to refer to malignant skin tumors, including melanoma and non-melanoma skin cancers, including primary and secondary skin cancers. In this regard, the term includes metastatic lesions arising from a primary skin cancer or metastasis to another cancer of the skin.

[0053] Among them, basal cell carcinoma and squamous cell carcinoma of the skin are examples of non-melanoma skin cancers. The term "basal cell carcinoma" used in this article refers to a low-grade malignant tumor originating from the basal cells of the skin. It includes nodular, superficial spreading, micronodular, mixed, etc. The term "squamous cell carcinoma of the skin" used in this article, also known as epidermoid carcinoma, is a malignant tumor originating from the squamous epithelial cells of the epidermis or adnexa, accounting for 20-50% of skin cancers. It is more invasive than basal cell carcinoma and is more prone to local and distant metastasis.

[0054] As used herein, the term "skin precancerous lesion" refers to an epithelial lesion or abnormal proliferation that is or may be a precursor to cancer, including but not limited to Bowen's disease, arsenic keratosis, radiation dermatitis, leukoplakia, or junctional nevus.

[0055] As used herein, the term "torso" refers to the body of a subject other than the arms, legs, or head.

[0056] As used herein, the phrase "until the skin cancer or its precancerous lesions are cleared" refers to a situation in which the lesions on a subject having skin cancer or its precancerous lesions are substantially or completely cleared from the treated area of ​​the subject. In one embodiment, "substantially" in this context means that more than 50% of the skin cancer or its precancerous lesions are cleared from the treated area of ​​the subject. In another embodiment, "substantially" means that more than 60% of the skin cancer or its precancerous lesions are cleared from the treated area of ​​the subject. In another embodiment, "substantially" means that more than 70% of the skin cancer or its precancerous lesions are cleared from the treated area of ​​the subject. In another embodiment, "substantially" means that more than 80% of the skin cancer or its precancerous lesions are cleared from the treated area of ​​the subject. In another embodiment, "substantially" means that more than 90% of the skin cancer or its precancerous lesions are cleared from the treated area of ​​the subject. In another embodiment, "substantially" means that more than 95% of the skin cancer or its precancerous lesions are cleared from the treated area of ​​the subject. In another embodiment, "substantially" means that more than 99% of the skin cancer or its precancerous lesions are cleared from the treated area of ​​the subject.

[0057] As used throughout this disclosure, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a method" includes a plurality of such methods and reference to "a dosage" includes reference to one or more dosages and equivalents thereof known to those skilled in the art, and so forth.

[0058] The term "comprising" is intended to indicate that the method includes the listed elements, but does not exclude other elements. When used to define a method, "consisting essentially of" shall mean excluding other elements that have any essential significance to the combination when used for the intended purpose. Thus, a method consisting essentially of the elements defined herein does not exclude substantial method steps. "Consisting of" means excluding multiple substantial method steps. Embodiments defined by each of these transition terms are within the scope of this application.

[0059] Unless expressly indicated otherwise, the terms "approximately" and "about" are synonymous. In one embodiment, "approximately" and "about" refer to ±5%, ±4.5%, ±4%, ±3.5%, ±3%, ±2.5%, ±2%, ±1.75%, ±1.5%, ±1.25%, ±1%, ±0.9%, ±0.8%, ±0.7%, ±0.6%, ±0.5% ±0.4%, ±0.3%, ±0.2%, ±0.1%, ±0.09%, ±0.08%, ±0.07%, ±0.06%, ±0.05%, ±0.04%, ±0.03%, ±0.02%, or ±0.01% of a stated amount, value, or duration. In another embodiment, "approximately" and "about" refer to ±2.5%, ±2%, ±1.75%, ±1.5%, ±1.25%, ±1%, ±0.9%, ±0.8%, ±0.7%, ±0.6%, ±0.5% of the listed amount, value, or duration. In another embodiment, "approximately" and "approximately" refer to ±1% of the listed amount, value, or duration. In another embodiment, "approximately" and "approximately" refer to ±0.5% of the listed amount, value, or duration. In another embodiment, "approximately" and "approximately" refer to ±0.1% of the listed amount, value, or duration.

[0060] The term "subject" includes any living organism that has skin cancer or a precancerous lesion thereof, or is at risk of developing skin cancer or a precancerous lesion thereof. In one embodiment, the term "subject" refers to a mammal that has skin cancer or a precancerous lesion thereof, or is at risk of developing skin cancer or a precancerous lesion thereof. In one embodiment, the term subject refers to a human that has skin cancer or a precancerous lesion thereof, or is at risk of developing skin cancer or a precancerous lesion thereof. Unless expressly indicated otherwise, the term "patient" is synonymous with "subject."

[0061] As used herein, the term "therapeutically effective amount" refers to the amount of an agent, such as 17β-xanthoside B, or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof, used to treat, improve, or prevent a confirmed disease or condition, such as skin cancer or its precancerous lesions, or to exhibit a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The exact effective amount for a subject depends on the subject's weight, size, and health; the nature and extent of the condition; and the therapeutic agent or combination of therapeutic agents selected for administration. The therapeutically effective amount for a given situation can be determined by routine experiments within the ability and judgment of a clinician.

[0062] For any compound, the therapeutically effective amount can be estimated in an animal model, typically rats, mice, rabbits, dogs or pigs. Animal models can also be used to determine suitable concentration ranges and routes of administration. Such information can then be used to determine useful doses and routes for administration in humans. Therapeutic / preventive efficacy and toxicity can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, such as ED50 (dose effective for 50% of the population treatment) and LD50 (dose lethal to 50% of the population). The dose ratio between toxicity and therapeutic effect is the therapeutic index, which can be expressed as the ratio of LD50 / ED50. The dosage can vary within this range, depending on the dosage form employed and the sensitivity of the subject.

[0063] Dosage and administration are adjusted to provide sufficient levels of the active ingredient or to maintain the desired effect. Factors that may be considered include the severity of the disease state, the location of the disease on the subject, the subject's overall health, the subject's age, weight and sex, diet, administration time and frequency, drug combination, reaction sensitivity and tolerance / response to treatment. Depending on the half-life and clearance rate, 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug can be administered daily, every other day, every three days, every four days, every five days, every six days, weekly, twice a week or once every two weeks.

[0064] For any of the methods described herein, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof can be administered topically, intradermally, intercutaneously, intragingivally, intraocularly, nasally, ocularly, transdermally, periodontally, subconjunctivally, sublingually, transmucosally, or otically. In one embodiment, 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof can be administered topically.

[0065] All percentages and ratios used herein are by weight unless otherwise specified.

[0066] The molecular formula of the term "17β-xanthoside B" used in this article is as follows:

[0067] Skin cancer is often caused by long-term sun exposure, ultraviolet radiation, exposure to biochemical carcinogens, and genetic factors. For example, squamous cell carcinoma tumor cells typically originate from the epidermis and irregularly infiltrate the dermis as the tumor grows. Therefore, specific delivery is required at the lesion site to meet the delivery requirements of the disease. The inventors of this application have discovered that 17β-xanthoside B or its pharmaceutically acceptable salts, racemates, stereoisomers, enantiomers, diastereomers, tautomers, metabolites, solvates, and prodrugs have a significant inhibitory effect on skin cancer tumor cells and can therefore be used to prepare drugs for the treatment of skin cancer.

[0068] It has been further discovered that 17β-xanthoside B or its pharmaceutically acceptable salts, racemates, stereoisomers, enantiomers, diastereomers, tautomers, metabolites, solvates, and prodrugs can penetrate the skin and, when administered topically to skin lesions, can significantly inhibit tumor cell growth. Thus, 17β-xanthoside B can be formulated into a topical drug that acts directly on skin lesions. Compared to oral or injectable drug formulations, topical formulations of 17β-xanthoside B can increase drug retention in the target skin and reduce the amount of drug entering the bloodstream, thereby reducing systemic adverse drug reactions.

[0069] 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug in the drug remains in a substantially dissolved form. Preferably, the drug contains at least 50% dissolved form, more preferably the drug contains at least 70% dissolved form, more preferably the drug contains at least 90% dissolved form, more preferably the drug contains 100% dissolved form. Undissolved crystals can exist in any known polymorphic form. As measured using laser diffraction, the particle size of 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug can vary between 1 micron and 100 microns (D90).

[0070] In some embodiments, the pharmaceutical composition is made of 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug and pharmaceutically acceptable excipients, so that the pharmaceutical composition is a drug suitable for external use.

[0071] The medicine provided according to the application can be in any suitable form, in the corresponding suitable form depending on the required purposes. For example, but not limited to, the medicine can be in the form of powders, sprays, ointments, pastes, creams, lotions, solutions (such as lotions, suspensions), patches, inhalants, gels (such as hydrogels) or emulsifiable pastes for direct external use. Wherein, depending on the form of use and purposes, suitable pharmaceutically acceptable adjuvants can be selected to provide the external medicine of corresponding form, wherein, pharmaceutically acceptable adjuvants include excipients, diluents, solubilizers, emulsifiers, antioxidants, adhesives, dispersants, lubricants, preservatives, etc.

[0072] In some embodiments, the pharmaceutical composition is a gel.

[0073] In some embodiments, the pharmaceutically acceptable excipients in the gel may optionally include a gel matrix. The gel has a large drug loading capacity, flexible dosing, long-term adhesion to the administration site, a long-lasting and mild effect, good biocompatibility and stability, and is easy to apply and clean.

[0074] In some embodiments, the gel matrix is ​​at least one selected from sodium carboxymethylcellulose, carbomer, gelatin, and alginate. Various types of carbomer can be used, such as carbomer 934, 940, 910, or mixtures thereof, with carbomer 940 being preferred. Carbomer can be commercially available, for example, from BF Goodrich. In some embodiments, pharmaceutically acceptable excipients in the gel may optionally include a humectant. Humectants can increase the adhesiveness of the formulation, improving its spreadability and viscosity.

[0075] Penetration enhancers can increase the rate at which active ingredients penetrate the skin and deliver the active ingredients to the desired site of action in the epidermis and / or dermis. Penetration enhancers can minimize skin irritation and systemic exposure, while achieving local skin / epidermal delivery and ensuring that most of the drug remains in the skin (i.e., the site of action) to treat specific skin conditions and reduce systemic toxic side effects.

[0076] As used herein, the term "weight %" refers to the weight of a component based on the total components, unless otherwise indicated.

[0077] In the embodiments of the present application, "treatment" refers to obtaining a desired pharmacological and / or physiological effect. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or may be therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, particularly humans, and includes inhibiting the disease, such as arresting the progression of the disease; or alleviating the disease, such as alleviating the symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a drug or compound to an individual to treat, cure, alleviate, improve, mitigate or inhibit the individual's disease, including but not limited to administering a drug containing the drugs described herein to an individual in need.

[0078] For the treatment of skin cancer or its precancerous lesions, the pharmaceutical compositions of the present application can be applied topically to the affected area in any conventional manner known in the art, for example, by a dropper, applicator stick or cotton swab, by an intradermal or transdermal patch, or simply by applying the formulation to the skin area with a finger, sponge, pad or swab.

[0079] The pharmaceutical composition can be used in combination with one or more other methods and drugs for treating skin cancer or its precancerous lesions. Within a certain time interval of administration of the pharmaceutical composition, it can be administered to a subject simultaneously or sequentially with other drugs or treatments so that the active ingredients or agents can work together to treat or prevent skin cancer or its precancerous lesions.

[0080] The compound contains an asymmetric center and can therefore exist as an enantiomer. The compound has multiple asymmetric centers, which can additionally exist as diastereomers. Enantiomers and diastereomers fall into the broader category of stereoisomers. All these possible stereoisomers include substantially pure resolved enantiomers, their racemic mixtures, and diastereomeric mixtures. All stereoisomers of the compound and / or their pharmaceutically acceptable salts are included. Unless otherwise stated, a reference to an isomer is applicable to any possible isomer. When an isomer component is not specifically indicated, all possible isomers are included.

[0081] Diastereomeric mixtures can be separated into individual diastereomers based on their physical chemical differences by methods well known to those skilled in the art, such as by chromatography and / or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with a suitable optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) each diastereomer to the corresponding pure enantiomer. Enantiomers can also be separated using a chiral HPLC column.

[0082] "Pharmaceutically acceptable salts" include, but are not limited to, salts of inorganic acids selected from, for example, hydrochlorides, phosphates, hydrogenphosphates, hydrobromides, sulfates, sulfites, and nitrates; and salts of organic salts selected from, for example, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate, benzoate, salicylate, stearate, alkanoates such as acetate, and salts of HOOC-(CH2)n-COOH, wherein n is selected from 0 to 4. Similarly, examples of pharmaceutically acceptable cations include, but are not limited to, sodium, potassium, calcium, aluminum, lithium, and ammonium salts.

[0083] Alternatively, if the compound is obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, an addition salt (e.g., a pharmaceutically acceptable addition salt) can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid, consistent with conventional procedures for preparing acid addition salts from basic compounds. Those skilled in the art will appreciate the various synthetic methods that can be used to prepare non-toxic, pharmaceutically acceptable addition salts without undue experimentation.

[0084] "Prodrug" refers to a compound that is converted into a parent drug in vivo. Prodrugs are often useful because, in some cases, they can be easier to administer than the parent drug. They can be bioavailable, for example, by oral administration, whereas the parent drug cannot. The prodrug can also have improved solubility in a pharmaceutical composition than the parent drug. An example of a prodrug is (but not limited to) a compound described herein that is administered as an ester (the "prodrug") to facilitate transport across the cell membrane, where water solubility is detrimental to this transfer, but which is then metabolically hydrolyzed into a carboxylic acid, an active entity, which, once inside the cell, is water-soluble. A further example of a prodrug can be a short peptide (polyamino acid) attached to an acid group, where the peptide is metabolized to reveal the active moiety. In certain embodiments, when administered in vivo, the prodrug is chemically converted into a biological, pharmaceutical, or therapeutically active form of the compound. In certain embodiments, the prodrug is enzymatically metabolized into a biological, pharmaceutical, or therapeutically active form of the compound by one or more steps or methods. To produce a prodrug, a pharmaceutically active compound is modified so that it is regenerated upon in vivo administration. Such prodrugs can be designed to alter the metabolic stability or transport properties of a drug, mask side effects or toxicity, thereby improving the drug's effect or altering other characteristics or properties of the drug. Once a pharmaceutically active compound is known, those skilled in the art can design a prodrug of the compound based on knowledge of pharmacodynamics and drug metabolism in vivo.

[0085] A "metabolite" of a compound disclosed herein is a derivative of the compound formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound formed when the compound is metabolized. As used herein, the term "metabolized" refers to the sum of processes by which a particular substance is altered by an organism (including but not limited to hydrolysis reactions and reactions catalyzed by enzymes, such as oxidation reactions). Thus, enzymes can produce specific structural transformations into compounds. For example, cytochrome P450 catalyzes various oxidation and reduction reactions, while diphosphoglucuronyltransferase catalyzes the conversion of activated glucuronic acid molecules to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines, and free sulfhydryl groups.

[0086] While particular embodiments of the present disclosure have been illustrated and described, various other changes and modifications may be made without departing from the spirit and scope of the present disclosure. It is intended that the scope of the appended claims include all such changes and modifications that are within the scope of the present disclosure.

[0087] Other features and advantages of the present application are apparent from the various examples provided. The examples provided illustrate various components and methods useful in implementing the present application.

[0088] These embodiments do not limit the claimed application. Based on this disclosure, a skilled artisan can identify and adopt other components and methods that can be used to implement the present application.

[0089] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become obvious from the description below, or will be learned through practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0090] Other features, objects and advantages of the present application will become more apparent upon reading the detailed description of non-limiting embodiments made with reference to the following drawings:

[0091] Figure 1 shows the effects of different concentrations of 17β-xanthoside B on the activity of TE.354.T cells (human skin basal cell carcinoma).

[0092] Figure 2 shows the effects of different drugs on the tumor volume of nude mice bearing SCL-1 cells (human skin squamous cell carcinoma cells): model control group, imiquimod cream group (50 mg / g), 17β-xanthoside B raw material group (1.6 mg / kg, administered once every other day by gavage), 17β-xanthoside B I, II, III (2 mg / g, 4 mg / g, 8 mg / g, administered once every other day to the skin), and group IV (4 mg / g, administered once a day to the skin). Each group was administered for 3 consecutive weeks.

[0093] Figure 3 shows the effects of different drugs on the tumor volume of COLO-16 cell (human skin squamous cell carcinoma)-bearing nude mice: model control group, imiquimod cream group (50 mg / g), 17β-xanthoside B raw material group (0.8 mg / kg, administered once every other day by gavage), 17β-xanthoside B I, II, III (2 mg / g, 4 mg / g, 8 mg / g, administered once every other day to the skin), and group IV (4 mg / g, administered once a day to the skin). Each group was administered for 3 consecutive weeks.

[0094] Figure 4 shows the effects of different drugs on the tumor size of COLO-16 cell (human skin squamous cell carcinoma)-bearing nude mice: model control group, imiquimod cream group (50 mg / g), 17β-xanthoside B raw material group (0.8 mg / kg, administered once every other day by gavage), 17β-xanthoside B I, II, III (2 mg / g, 4 mg / g, 8 mg / g, administered once every other day to the skin), and group IV (4 mg / g, administered once a day to the skin). Each group was administered for 3 consecutive weeks. DETAILED DESCRIPTION

[0095] The present invention will be further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are only used to explain the relevant invention, rather than to limit the invention.

[0096] It should be noted that, unless there is any conflict, the embodiments and features in the embodiments of this application can be combined with each other.

[0097] It should be noted that the endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be considered to be specifically disclosed herein.

[0098] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. All reagents or instruments used without specifying the manufacturer are commercially available conventional products.

[0099] Example 1

[0100] This example investigates the inhibitory effect of 17β-xanthoside B on TE.354.T, A431, SCL-1, and COLO-16 cancer cells. The experimental results show that it has a strong inhibitory effect on these cancer cells. The experimental process is as follows:

[0101] The passaged cells were cultured in a 37°C, 5% CO2 saturated humidity incubator using complete medium containing 10% FBS, and the cell density was adjusted to 3-5×10 4 / mL, 100 μL of cell suspension was added to each well of a 96-well culture plate and cultured overnight (approximately 16-24 hours) in a 37°C, 5% CO2 saturated humidity incubator. Test substance dose groups and a vehicle control group were established. The next day, the original culture medium was discarded, and 100 μL / well of 20% FBS complete medium was added, followed by 100 μL / well of the test substance or vehicle, with a final FBS concentration of 10%. A negative control group was also established: 100 μL of 20% FBS complete medium + 100 μL of incomplete medium were added to each well, with a final FBS concentration of 10%, containing cells. A zero adjustment group (background) was established: 200 μL of culture medium alone (100 μL of 20% FBS complete medium + 100 μL of incomplete medium) was added, without drug or cells. Four replicates were performed for each concentration group.

[0102] After culturing for 48 hours according to the test conditions, the supernatant was discarded and 110 μL of CCK-8 working solution (100 μL of 10% FBS complete medium and 10 μL of CCK-8 solution) was added to each well. After further culturing for 4 hours, the optical density (A450) of each well at a wavelength of 450 nm was measured using a microplate reader. The mean value (T) of each drug-treated group well was compared with the mean value (C) of the negative control well. The percentage inhibition rate of each concentration group was calculated using the following formula [inhibition rate = (1-T / C) × 100%], and the half-maximal inhibitory concentration (IC) was calculated. 50 The specific experimental results are shown in the following table:

[0103] TE.354.T: TE.354.T cells were cultured and passaged in a complete medium containing 15% FBS in a 37°C, 5% CO2 saturated humidity incubator. The cell density was adjusted to 1×10 4 / mL, add 1mL of cell suspension to each well of a 24-well culture plate and incubate overnight (approximately 16-24 hours) in a humidified incubator at 37°C and 5% CO2. Dose groups of test substance, a positive control group (cisplatin), and a vehicle control group were established. The next day, 1μL / well of test substance at various concentrations and 2μL / well of 5mg / mL of the positive control (cisplatin) were added. Cells were cultured according to experimental conditions, and daily photographs were taken to record cell growth and survival. After 72 hours, the supernatant was discarded, and 0.5mL of CCK8 working solution (containing 50μL of CCK8 stock solution) was added to each well. After incubation in a cell culture incubator for 2-3 hours, the optical density (A450) of each well at a wavelength of 450nm was measured using a microplate reader. The values ​​(T) of each drug-treated well were compared with the values ​​(C) of the negative control wells. The percent inhibition rate of each concentration group was calculated using the following formula: [Inhibition rate = (1-T / C) × 100%], and the half-maximal inhibitory concentration (IC50) was determined. The results are shown in FIG1 , which indicate that the half-maximal inhibitory concentration obtained by the assay was approximately 10 ng / mL, demonstrating a high activity in inhibiting cancer cell proliferation.

[0104] Example 2

[0105] 17β-xanthoside B is poorly soluble in water, and its solubility in water is about 70μg / mL. Therefore, the solubility of 17β-xanthoside B in organic solvents of different concentrations was investigated. The experimental results are shown in the following table:

[0106] It can be seen that 17β-xanthoside B has good solubility in ethanol and propylene glycol, and can be used as solvents for dissolving 17β-xanthoside B.

[0107] Example 3

[0108] The solubility of 17β-xanthoside B in ethanol at different concentrations (20℃) was studied.

[0109] Weigh an appropriate amount of 17β-glucosidoside B raw material into a beaker, a total of 3 parts. Dissolve the first part in anhydrous ethanol first, then dilute with water to a final ethanol concentration of 20%. The sample concentration is 8 mg / g, and observe whether there is precipitation. Dissolve the second part in anhydrous ethanol first, then dilute with water to a final ethanol concentration of 75%, 66%, 50% and 40%, respectively. The corresponding sample concentrations are 50, 40, 32 and 24 mg / g, respectively, and observe whether there is precipitation. Dissolve the third part in anhydrous ethanol, then dilute with water to an ethanol concentration of 45%, and the sample concentration is 20 mg / g. Observe whether there is precipitation. The results are shown in the following table:

[0110] It can be seen that 8 mg / g of 17β-glucosidoside B can be dissolved in 20% ethanol; 20 mg / g of 17β-glucosidoside B can be dissolved in 45% ethanol.

[0111] Example 4

[0112] Compatibility test of sodium carboxymethylcellulose and carbomer with 17β-xanthoside B:

[0113] After 17β-glucosidoside B was mixed with sodium carboxymethyl cellulose and carbomer in a mass ratio of 1:5, the mixture was placed at a temperature of 60℃±2℃, a relative humidity of 90%±5% for 10 days and under conditions of cumulative total illumination greater than 1.2×106Lux·hr to investigate the effect of the gel matrix on the stability of 17β-glucosidoside B. The experimental results showed that 17β-glucosidoside B and sodium carboxymethyl cellulose had no effect on the relevant substances and content under high temperature, high humidity and light conditions, and its stability was significantly better than that of carbomer.

[0114] The compatibility test results of 17β-glucosidoside B and sodium carboxymethyl cellulose are shown in the following table.

[0115] Example 5

[0116] Study the effect of different contents of gel matrix on the quality of the gel of this application:

[0117] Weigh 17β-xanthoside B raw material, add anhydrous ethanol and stir ultrasonically to dissolve; add azone and stir evenly; add water and stir evenly; add sodium carboxymethyl cellulose and stir evenly; add glycerol and stir evenly, and swell overnight; homogenize to obtain the finished gel. The physical properties of each gel were scored. The experimental results are shown in the following table:

[0118] A total score of 1-10 indicates that the physical properties of the prepared gel are poor, which may affect the patient's medication experience. A total score of 11-15 indicates that the gel is suitable for topical administration and provides a good medication experience. A total score of 16-20 indicates that the gel has good physical properties and is easy to administer without any discomfort. It can be seen that the gel obtained when the sodium carboxymethyl cellulose content is within the range of 1.0-3.0% by weight has good physical properties.

[0119] Example 6

[0120] The gel preparation was prepared according to the method of Example 5, and the effect of different contents of moisturizer on the quality of the gel preparation of the present application was studied. The experimental results are shown in the following table:

[0121] It can be seen that when the glycerol content is in the range of 15-25 wt %, the obtained gel has good physical properties.

[0122] Example 7

[0123] Preparation of gel:

[0124] Take 25 ml of distilled water and 12.5 ml of glycerol and mix them evenly, add 17β-glucosidoside B and 10 ml of ethanol, and dissolve them by ultrasonication.

[0125] Dissolve the prescribed amount of ethylparaben in 2 ml of ethanol, stir evenly, add to the above solution, and stir evenly.

[0126] Add the prescribed amount of CMC-Na to the above mixed solution, stir until it becomes uniform, add water to 50g, and stir evenly to obtain the product.

[0127] Example 8

[0128] Gels were prepared according to the method of Example 5, and the skin irritation of gels containing different penetration enhancers and different amounts of penetration enhancers was studied:

[0129] Healthy young pigs with no back injuries were used, half male and half female. The left and right sides of the same pigs were used for drug administration. Transdermal patch administration was performed, with the test gel administered on the left side and white vaseline administered on the right side as a negative control. The administration area was 3.0 cm × 3.0 cm, and the dosage was 20 mg / cm. 2 The drug was administered once daily, with the application continued on both sides of the skin for approximately 4 hours. The application site was then cleaned with warm 0.9% sodium chloride injection. A single dose was administered. Erythema and edema at the application site were observed and recorded before administration, and 1 hour, 24 hours, 48 ​​hours, 72 hours, and 14 days after drug removal. Erythema and edema were scored according to the standard. All tests met the irritation requirements.

[0130] Among them, the test gel contains 1.2% by weight of 17β-xanthoside B, 1.5% by weight of sodium carboxymethyl cellulose, 40% by weight of anhydrous ethanol, 0.5-1.0% by weight of a permeation enhancer, 20% by weight of glycerol and the balance of water. The permeation enhancers of 0.5-1.0% by weight are 1.0% azone, 1.0% borneol, 0.75% borneol and 0.5% borneol respectively.

[0131] Example 9

[0132] The Franz diffusion cell method was used to determine the in vitro transdermal absorption, including the transdermal amount, epidermal residual amount and intradermal retention amount.

[0133] Transdermal dose: Pigskin (soaked in physiological saline for about 30 minutes before use and dried with filter paper) was fixed between the receiving cell and the dosing cell, with the stratum corneum facing the dosing cell. Approximately 200 mg of the gel of the present application was taken, accurately weighed, and evenly applied to the pigskin on one side of the dosing cell. 18 mL of 15% ethanol-physiological saline preheated to 32°C ± 0.5°C was added to the receiving cell and stirred at 200 rpm. 0.5 mL of the receptor solution was taken at 1, 2, 3, 4, and 6 hours (a blank receptor solution of the same volume and temperature was added to the receiving cell at the same time). The solution was centrifuged, the supernatant was taken, and HPLC determination was performed. The transdermal drug amount was calculated by the peak area.

[0134] Epidermal residue: Take pig skin after 6 hours of penetration of this product (note that the gel remaining on the surface has flowed out), wash it with 25 mL of saturated sodium chloride solution 5 times (5 mL each time, vortex for 0.5 minute), place the washing solution in a 100 mL volumetric flask, add 25 mL of methanol, and extract by ultrasonic for 20 minutes. Dilute to the scale with 50% methanol-saturated sodium chloride solution, shake well, filter, take the filtrate, and perform HPLC determination. The epidermal residue is calculated by the peak area.

[0135] Intradermal retention: Take the rinsed pig skin, add 25 mL of 80% methanol-saturated sodium chloride solution, heat in a 70°C water bath for 30 minutes, chop, homogenize, centrifuge (4°C, 14000 rpm for 10 minutes), take the supernatant, filter, take the filtrate, perform HPLC determination, and calculate the residual amount by peak area.

[0136] The test gel contains 0.4 wt% of 17β-glucosidoside B, 1.5 wt% of sodium carboxymethyl cellulose, 40 wt% of anhydrous ethanol, 0.5 wt% of borneol, 20 wt% of glycerol and the balance of water, and is prepared according to the method of Example 5.

[0137] Calculations show that the drug in the gel of the present invention penetrates less than 1% of the skin within 6 hours, with an epidermal residual amount of 80-95% and an intradermal retention of approximately 10%. This indicates that the drug in the gel prepared by the present invention is essentially retained in the skin, resulting in a high local drug concentration and preventing absorption into the bloodstream, thereby reducing side effects associated with systemic drug distribution.

[0138] Example 10

[0139] Healthy, SPF-grade Hartley guinea pigs with intact dorsal skin were selected, half male and half female. A negative control (white petrolatum), a positive control (2,4-dinitrochlorophenyl petrolatum ointment), a test group (17β-xanthoside B gel, 2 mg / g of 17β-xanthoside B), a test group (17β-xanthoside B gel, 4 mg / g of 17β-xanthoside B), and a test group (17β-xanthoside B gel, 8 mg / g of 17β-xanthoside B) were set up. Each group received 0.5 g of the drug per dose, and the skin application area was 2.5 cm × 2.5 cm. Each animal in each group was exposed to the left dorsal skin for 6 hours. Skin reactions were observed and scored before drug administration and 1 and 24 hours after cleaning the residue. Sensitization was performed three times, once on the 1st, 8th, and 15th days of administration. On the 29th day of administration, the drug was placed in contact with the right side of the animal's skin for 6 hours. The skin reaction was observed and recorded before administration and 24 and 48 hours after cleaning the residue. The test results were then read.

[0140] The test gels of test group 1, group 2 and group 3 respectively contained 1.5 wt% sodium carboxymethyl cellulose, 40 wt% anhydrous ethanol, 0.5 wt% borneol, 20 wt% glycerol and the balance water, and were prepared according to the method of Example 5.

[0141] The results showed that the gel preparations of the present application did not cause allergic reactions in guinea pig skin, and the allergic reactions were all negative.

[0142] Example 11

[0143] Healthy New Zealand rabbits with no dorsal injuries were used, half male and half female. A blank gel control group and a 17β-xanthoside B gel group (17β-xanthoside B content of 2 mg / g and 4 mg / g, respectively) were set up. The test substance was administered to the left side of the rabbits, while white petrolatum was administered to the right side as a negative control. The administration area was 3.0 cm × 3.0 cm. After the application was continued for approximately 4 hours on both sides of the skin, the administration site was cleaned with warm 0.9% sodium chloride injection. A single dose was administered. Erythema and edema at the administration site were observed and recorded before administration and 1 hour, 24 hours, 48 ​​hours, 72 hours, and 14 days after drug removal. Erythema and edema were scored according to the scoring criteria. Histopathological examinations were performed on the left side of the skin at the administration site in each group 72 hours and 14 days after drug removal.

[0144] The results showed that when the blank gel control group and the 17β-xanthoside B gel group (2 mg / g, 4 mg / g) were administered percutaneously to the intact skin of New Zealand rabbits, the skin irritation reaction scores on the left and right sides of each animal were within the non-irritating range, and no abnormalities were found in the microscopic examination results, and no skin irritation reaction related to the test substance was observed.

[0145] The test gel contains 1.5 wt % sodium carboxymethyl cellulose, 40 wt % anhydrous ethanol, 0.5 wt % borneol, 20 wt % glycerol and the balance water, and is prepared according to the method of Example 5.

[0146] Example 12

[0147] Quarantine-qualified male BALB / c-nu nude mice weighing 13.4-20.5 g were inoculated with 3×10 SCL-1 cells in 0.2 mL of logarithmic growth phase. 6 cells / mL (matrigel and cell suspension were mixed in a ratio of 1:2) and inoculated into the right armpit of nude mice until the average tumor volume of the nude mice reached 100 mm 3 At about 14 days, 84 tumor-bearing nude mice were randomly divided into 7 groups according to tumor volume (intact tumor, no ulceration), including model control group, imiquimod cream group (50 mg / g), 17β-xanthoside B raw material group (1.6 mg / kg, administered orally every other day), 17β-xanthoside B I, II, III (2 mg / g, 4 mg / g, 8 mg / g, administered to the skin every other day), and IV group (4 mg / g, administered to the skin once a day). The model control group, imiquimod cream group, and 17β-xanthoside B I, II, III, and IV groups were treated with 0.01 g gel or cream / cm 2 Drug administration was performed via transdermal application at the tumor site, with each drug contact period lasting 4 hours. Each group received drug administration for 3 consecutive weeks. Every 3 days during the administration period, the major diameter (a) and minor diameter (b) of the tumor were measured with a vernier caliper. Tumor volume (TV), relative tumor volume (RTV), and relative tumor growth rate (T / C, %) were calculated. Tumor tissue was dissected and weighed after the last dose, and the tumor growth inhibition rate was calculated.

[0148] Among them, 17β-xanthoside B gel groups I, II, III, and IV respectively contain 1.5% by weight of sodium carboxymethyl cellulose, 40% by weight of anhydrous ethanol, 0.5% by weight of borneol, 20% by weight of glycerol, the remainder being water and the corresponding 17β-xanthoside B, and are prepared according to the method of Example 5.

[0149] The experimental results are shown in Figure 2. It can be seen that the tumor inhibition rate of 17β-xanthoside B raw material (1.6 mg / kg) after 21 days of administration is greater than 70%, and the tumor inhibition rates of 17β-xanthoside B gel groups I, II, III, and IV are greater than 60%, 80%, 95%, and 85%, respectively. It has a significant tumor inhibition effect, and the tumor inhibition effect is significantly better than that of imiquimod cream. It is suitable for preparing a topical drug to act on skin lesions for the treatment of skin cancer, while improving the therapeutic effect and reducing the systemic adverse reactions of the drug.

[0150] Example 13

[0151] Male BALB / c-nu nude mice that passed quarantine, weighing 13.7-21.1 g, were inoculated with 0.2 mL of COLO-16 cells in the logarithmic growth phase (6.7 × 10 6 The cells were inoculated into the right armpit of nude mice until the average tumor volume of the nude mice reached 100 mm 3 At about 14 days, 56 tumor-bearing nude mice were randomly divided into 7 groups according to tumor volume (intact tumor, no rupture), including model control group, imiquimod cream group (50 mg / g), 17β-xanthoside B raw material group (0.8 mg / kg, oral administration once every other day), 17β-xanthoside B I, II, III (2 mg / g, 4 mg / g, 8 mg / g, skin administration once every other day), and IV group (4 mg / g, skin administration once a day). The model control group, imiquimod cream group, and 17β-xanthoside B I, II, III, and IV groups were treated with 0.01 g gel or cream / cm 2 Drug administration was performed via transdermal application at the tumor site, with each drug contact period lasting 4 hours. Each group received drug administration for 3 consecutive weeks. Every 3 days during the administration period, the major diameter (a) and minor diameter (b) of the tumor were measured with a vernier caliper. Tumor volume (TV), relative tumor volume (RTV), and relative tumor growth rate (T / C, %) were calculated. Tumor tissue was dissected and weighed after the last dose, and the tumor growth inhibition rate was calculated.

[0152] Among them, gel groups I, II, III, and IV respectively contain 1.5% by weight of sodium carboxymethyl cellulose, 40% by weight of anhydrous ethanol, 0.5% by weight of borneol, 20% by weight of glycerol, the remainder being water, and the corresponding amounts of 17β-glucosidoside B in groups I, II, III, and IV, and are prepared according to the method of Example 5.

[0153] The experimental results are shown in Figures 3-4. It can be seen that the tumor inhibition rate of 17β-xanthoside B raw material (0.8 mg / kg) after 21 days of administration is greater than 80%, and the tumor inhibition rates of 17β-xanthoside B gel groups I, II, III, and IV are greater than 70%, 90%, 99%, and 99%, respectively. It has a significant tumor inhibition effect, and the tumor inhibition effect is significantly better than that of imiquimod cream. It is suitable for preparing a topical drug to act on skin lesions for the treatment of skin cancer, while improving the therapeutic effect and reducing the systemic adverse reactions of the drug.

[0154] Example 14

[0155] A375.S2 human malignant melanoma cells were subcutaneously inoculated in the right axilla of nude mice until the average tumor volume reached 100 mm. 3 When the rats were treated with the drug, they were divided into model control group, gel group 1 (0.5% borneol), gel group 2 (1.5% borneol), and gel group 3 (1.0% azone). There were 6 animals in each group, all male. The drug was administered through skin application, and the administration site was the tumor site. The test substance was applied to the corresponding administration area of ​​the animal's administration site, covered with a layer of plastic film, then covered with two layers of gauze, and fixed with non-irritating paper tape. The drug was administered once a day for 2 consecutive weeks, and the dosage was 0.2g / cm 2 The tumor diameters of the animals were measured and the relative tumor growth rate (T / C) was calculated. The nude mice were sacrificed by cervical dislocation and the tumors were removed and weighed to calculate the tumor inhibition rate.

[0156] Among them, gel groups 1, 2, and 3 respectively contain 0.2% by weight of 17β-glucosidoside B, 1.5% by weight of sodium carboxymethyl cellulose, 40% by weight of anhydrous ethanol, 20% by weight of glycerol, the remainder of water, and the corresponding penetration enhancer, and are prepared according to the method of Example 5.

[0157] The experimental results are shown in the following table.

[0158] It can be seen that applying the gel (2 mg / g) containing 0.5% borneol and 1.5% borneol once a day has an anti-tumor effect on nude mice bearing human malignant melanoma cells (A375.S2); applying the gel (2 mg / g) containing 1.0% azone once a day has an anti-tumor effect on nude mice bearing human malignant melanoma cells (A375.S2).

[0159] Experiments have shown that the tumor inhibition rate of 17β-xanthoside B on SCL-1 cells and COLO-16 cells is better than that on A375.S2 human malignant melanoma cells.

[0160] Example 15

[0161] Pharmaceutical composition for clinical treatment of non-melanoma skin cancer and Bowen's disease

[0162] Clinical studies were conducted to evaluate the activity and safety of topical 17β-xanthoside B in subjects with non-melanoma skin cancer and Bowen's disease. 17β-xanthoside B demonstrated promising, clinically relevant activity in the treatment of non-melanoma skin cancer and Bowen's disease. 17β-xanthoside B was safe based on local tolerability and local skin reactions (LSRs).

[0163] This trial consists of a Phase I (single-dose ascending trial) and a Phase II (expansion trial). The Phase I trial is a multicenter, open-label, dose-escalation trial that investigates the safety, tolerability, and PK characteristics of different doses of 17β-xanthoside B gel applied to the lesions in Chinese patients with non-melanoma skin cancer (including basal cell carcinoma and squamous cell carcinoma of the skin) and Bowen's disease, and preliminarily investigates the efficacy of 17β-xanthoside B. The Phase II trial is a multicenter, open-label trial that investigates the efficacy, safety, and PK characteristics of 17β-xanthoside B gel at the target dose in Chinese subjects with non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma of the skin) and Bowen's disease.

[0164] 1. Case selection

[0165] Patients aged ≥18 years were selected. All patients cooperated with the study and provided informed consent.

[0166] 2. Inclusion criteria

[0167] Inclusion criteria: ① patients met the diagnostic criteria for cutaneous squamous cell carcinoma, basal cell carcinoma, and Bowen's disease in Clinical Dermatology and were diagnosed by histopathological examination or non-invasive diagnosis (dermoscopy, skin CT, etc.); ② complete clinical data.

[0168] 3. Methods

[0169] The first phase (single ascending dose trial) included single and multiple dosing. After single dosing, DLT was observed for 3 days. On the 4th day, multiple dosing was started, with dosing every other day for 3 consecutive weeks.

[0170] Dosage: Apply the gel evenly to the affected area, avoiding rinsing and friction. Cover with a breathable, waterproof dressing and secure with a non-irritating adhesive tape. Ensure the drug remains in contact with the skin for 4 ± 0.5 hours between doses. To remove the drug, cleanse the application area with room-temperature saline.

[0171] Dosage calculation: Dosage area (cm 2 ) = square of the radius of the affected area * π

[0172] Dosage (gel) mg = current dose group per cm 2 Dosage (gel dosage)*dosage area

[0173] The conversion formula between mg and microliters is: gel weight ÷ gel density = volume (corresponding to microliters ul)

[0174] Dosing area: The dosing area is 2 treatment units (treatment unit: the area of ​​one index finger joint of each subject is one treatment unit). The total dosing area of ​​the target lesion does not exceed 25cm 2 .

[0175] Medication administration during hospitalization must be under the supervision of a physician or nurse, and the dosage and schedule must be recorded in the subject's diary card. During non-hospitalized periods, the study physician and nurse should instruct the subject to take medication according to the prescribed dosing schedule at a fixed time each day. For detailed dosing schedules, please refer to the Standard Operating Procedures for Medication Administration.

[0176] The frequency of administration of the trial drug and the setting of the interval period may be adjusted appropriately based on human safety and PK data as well as efficacy.

[0177] 4. Observation indicators

[0178] Evaluate clinical effects.

[0179] Efficacy evaluation indicators applicable to the cohorts of subjects with basal cell carcinoma of the skin, squamous cell carcinoma of the skin, and Bowen's disease:

[0180] CR: complete remission, indicating that all tumor target lesions disappear;

[0181] PR: partial response, indicating a reduction in tumor volume of at least 30%;

[0182] SD: stable disease, indicating no significant change in tumor volume;

[0183] Objective response rate (ORR): The proportion of subjects with CR and PR (calculated based on the best response during the dosing period) in the corresponding analysis set;

[0184] Disease control rate (DCR): the proportion of subjects with CR, PR and SD (calculated based on the best response during the dosing period) in the corresponding analysis set;

[0185] Duration of response (DOR): refers to the time from the first assessment of CR or PR to the first assessment of PD or death from any cause;

[0186] Time to tumor progression (TTP): refers to the time from the first dose of the subject to the date of clear disease progression;

[0187] Progression-free survival (PFS): refers to the time from the first medication of the subject to the date of clear disease progression or death due to any cause;

[0188] Dermatology Life Quality Index: SF-36 scoring scale evaluates the subjects' quality of life;

[0189] Safety indicators: adverse events (including local reactions at the medication site), physical examination, vital signs, laboratory tests (including blood routine, blood biochemistry, urine routine, etc.), electrocardiogram, etc.

[0190] 5. The following is a summary of the safety and efficacy data of the subjects enrolled in the first phase

[0191] A total of 29 subjects were enrolled in this study. The demographic information of the subjects is shown in the table below.

[0192] Summary demographic data – All enrolled subjects (China and Australia)

[0193] Safety results

[0194] Overall, 29 subjects received at least one topical application of 17β-xanthoside B gel, with the doses in the five groups being 0.008 mg / cm 2 , 0.016mg / cm 2 , 0.027mg / cm 2 , 0.04mg / cm 2 , 0.06mg / cm 2 , and exhibited a good safety profile. In terms of safety, 17β-xanthoside B topical gel performed similarly in Chinese and Australian subjects. According to the researchers' evaluation, drug-related adverse events were mainly skin administration site reactions.

[0195] A total of 20 subjects (69%) experienced treatment-emergent adverse events (TRAEs) assessed by the investigators as drug-related. The most commonly reported TRAEs were pruritus (7 / 29, 24.1%), desquamation (5 / 29, 17.2%), and crusting (5 / 29, 17.2%), primarily mild local reactions at the administration site. All AEs were grade 1 or 2 in severity, and no TEAEs of CTCAE grade 3 or higher occurred.

[0196] No SAEs occurred and no DLTs were observed.

[0197] No AEs leading to death occurred.

[0198] In summary, the safety profile of 17β-xanthoside B was similar in Chinese and Australian subjects. According to the researchers' evaluation, drug-related adverse events were primarily local reactions at the skin administration site, such as pruritus, skin peeling, and skin scabs. These were all mild reactions of grade 1-2, indicating a generally good safety profile.

[0199] Effectiveness Results:

[0200] Based on the safety, tolerability, pharmacokinetic and efficacy data generated in this clinical study, the subjects who were evaluable for efficacy in dose groups 1-5 (China and Australia) of this study included 6 patients with basal cell carcinoma of the skin, 1 patient with non-in situ squamous cell carcinoma of the skin, and 6 patients with Bowen's disease.

[0201] Efficacy of 17β-xanthoside B in the treatment of basal cell carcinoma of the skin

[0202] In the fifth dose group, a total of 6 patients with basal cell carcinoma of the skin whose efficacy could be evaluated at D16, D24 and 28 days after the last dose were enrolled. The results based on the investigator's assessment (according to the RECIST1.1 standard) showed that at D16, D24 and 28 days after the last dose, the best efficacy evaluation of the three subjects was PR, the disease control rate (DCR) of 17β-xanthoside B in the treatment of BCC was 100%; the ORR was 50%.

[0203] Efficacy of 17β-xanthoside B in the treatment of cutaneous squamous cell carcinoma

[0204] In the 5th dose group, a patient with cutaneous squamous cell carcinoma whose efficacy was evaluable at D16, D24 and 28 days after the last dose was enrolled. The results based on the investigator's assessment (according to the RECIST1.1 standard) showed that the subject's best efficacy assessment was PR at D16, D24 and 28 days after the last dose was followed up.

[0205] The efficacy of 17β-flavoglucosidase B in the treatment of Bowen's disease

[0206] Six patients with Bowen's disease whose efficacy was evaluable were enrolled in the dose-escalation phase of this study. Based on investigator assessment (according to RECIST 1.1 criteria), the disease control rate (DCR) of 17β-xanthoside B in Bowen's disease was 100%, the complete response rate (CRR) was 16.67%, and the overall response rate (ORR) was 66.7%.

[0207] Pharmacokinetic study results:

[0208] Serum concentrations of the topical gel were measured and analyzed in Chinese and Australian patients enrolled in the dose escalation phase of the clinical study (dose levels 1-5).

[0209] The serum concentration of 17β-xanthoside B was determined by liquid chromatography-mass spectrometry (LC / MS / MS), and the lower limit of quantification (LLOQ) was 0.02 ng / mL. 2 No increase in systemic exposure of 17β-xanthoside B was observed with increasing dose level across the dose range. Due to the low overall systemic absorption rate and sparse data, a statistically significant pharmacokinetic evaluation could not be performed (only seven Chinese subjects had plasma concentrations above the LLOQ, ranging from 0.022 to 0.106 ng / mL, and four Australian subjects had plasma concentrations above the LLOQ, ranging from 0.021 to 0.101 ng / mL).

[0210] It can be seen that the gel of the present application has a good therapeutic effect and is proven to have good safety.

[0211] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0212] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations on the present application. Ordinary technicians in this field can modify, replace and vary the above embodiments within the scope of the present application.

Claims

1. A compound, which is 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof, and is used for external administration to treat and / or prevent skin cancer or precancerous lesions thereof.

2. A method for treating and / or preventing skin cancer or its precancerous lesions, comprising administering 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug.

3. The method according to claim 2, characterized in that The method comprises administering a therapeutically effective amount of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, or prodrug thereof to a subject in need thereof.

4. The method according to any one of the preceding claims, characterized in that The skin cancer includes melanoma and / or non-melanoma skin cancer.

5. The method according to any one of the preceding claims, characterized in that The non-melanoma skin cancer is basal cell carcinoma and / or squamous cell carcinoma of the skin.

6. The method according to any one of the preceding claims, characterized in that The skin precancerous lesions include epithelial lesions or abnormal proliferation, preferably the precursor of cancer or can be the precursor of cancer, more preferably Bowen's disease, arsenic keratosis, radiation dermatitis, mucosal leukoplakia or junctional nevus.

7. The method according to any one of the preceding claims, characterized in that 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is administered to the affected area of ​​the subject at the following dosages: about 0.0003 mg to about 100 mg; about 0.008 mg to about 90 mg; about 0.1 mg to about 80 mg; about 0.1 mg to about 70 mg; about 0.1 mg to about 60 mg; about 0.1 mg to about 50 mg; about 0.1 mg to about 40 mg; about 0.1 mg to about 30 mg; about 0.1 mg to about 20 mg; about 0.1 mg to about 15 mg; about 0.1 mg to about 10 mg; about 0.1 mg to about 8 mg; about 0.2 mg to about 5 mg; about 0.5 mg to about 2.5 mg; about 0.02 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, About 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.6 mg, about 1.7 mg, about 1.8 mg, about 1.9 mg, about 2.0 mg, about 2.1 mg, about 2.2 mg, about 2.3 mg, about 2.4 mg, about 2.5 mg, about, 2.6 mg, about 2.7 mg, about 2.8 mg, about 2.9 mg, about 3 mg, about 4 mg, or about 5 mg.

8. The method according to any one of the preceding claims, characterized in that 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is administered to the affected area of ​​the subject at the following dosage: about 0.0003 mg / cm 2 To about 10mg / cm 2 ; preferably about 0.001 mg / cm 2 About 1mg / cm 2 ; preferably about 0.002mg / cm 2 To about 0.5mg / cm 2 ; preferably about 0.004 mg / cm 2 To about 0.25mg / cm 2 ; preferably about 0.008 mg / cm 2 To about 0.2mg / cm 2 ; preferably about 0.01 mg / cm 2 To about 0.18mg / cm 2 ; preferably about 0.016mg / cm 2 To about 0.16mg / cm 2 ; preferably about 0.02mg / cm 2 To about 0.15mg / cm 2 ; preferably about 0.025mg / cm 2 To about 0.14mg / cm 2 ; preferably about 0.027mg / cm 2 To about 0.13mg / cm 2 ; preferably about 0.03mg / cm 2 To about 0.125mg / cm 2 ; preferably about 0.035mg / cm 2 To about 0.12mg / cm 2 ; preferably about 0.04mg / cm 2 To about 0.1mg / cm 2 ; about 0.001mg / cm 2 , about 0.002mg / cm 2 , about 0.003mg / cm 2 , about 0.004mg / cm 2 , about 0.005mg / cm 2 , about 0.006mg / cm 2 , about 0.007mg / cm 2 , about 0.008mg / cm 2 , about 0.009mg / cm 2 , about 0.01mg / cm 2 , about 0.015mg / cm 2 , about 0.02mg / cm 2 , about 0.025mg / cm 2 , about 0.03mg / cm 2 , about 0.035mg / cm 2 , about 0.04mg / cm 2 , about 0.045mg / cm 2 , about 0.05mg / cm 2 , about 0.055mg / cm 2 , about 0.06mg / cm 2 , about 0.065mg / cm 2 , about 0.07mg / cm 2 , about 0.075mg / cm 2 , about 0.08mg / cm 2 , about 0.085mg / cm 2 , about 0.09mg / cm 2 , about 0.095mg / cm 2 , about 0.1mg / cm 2 , about 0.15mg / cm 2 , about 0.2mg / cm 2 , about 0.25mg / cm 2 , about 0.3mg / cm 2 , about 0.35mg / cm 2 , about 0.4mg / cm 2 , about 0.45mg / cm 2 , about 0.5mg / cm 2 , about 0.55mg / cm 2 , about 0.6mg / cm 2 , or about 0.65 mg / cm 2 .

9. The method according to any one of the preceding claims, characterized in that The affected area in this subject was approximately 0.01 cm 2 About 300cm 2 Preferably, the diseased area of ​​the subject is about 1 cm 2 To about 200cm 2 , about 1cm 2 To about 100cm 2 , about 1cm 2 To about 75cm 2 , about 1cm 2 To about 50cm 2 , about 1cm 2 To about 25cm 2 ; About 10cm 2 To about 200cm 2 , about 10cm 2 To about 100cm 2 , about 10cm 2 To about 75cm 2 , about 10cm 2 To about 50cm 2 , about 10cm 2 To about 25cm 2 ; About 25cm 2 To about 200cm 2 、Approx. 25cm 2 To about 100cm 2 、Approx. 25cm 2 To about 75cm 2 ; About 25cm 2 To about 90cm 2 、Approx. 25cm 2 To about 80cm 2 , or about 25cm 2 To about 70cm 2 、Approx. 25cm 2 About 60cm 2 、Approx. 25cm 2 To about 50cm 2 、Approx. 25cm 2 To about 40cm 2 , or about 25cm 2 About 30cm 2 Preferably, the diseased area of ​​the subject is about 0.2 cm 2 、About 0.5cm 2 , about 1cm 2 , about 2cm 2 , about 3cm 2 、Approx. 4cm 2 、About 5cm 2 、About 6cm 2 、About 7cm 2 、About 8cm 2 、About 9cm 2 , about 10cm 2 、Approx. 11cm 2 、Approx. 12cm 2 、Approx. 13cm 2 、Approx. 14cm 2 、Approx. 15cm 2 、Approx. 16cm 2 、Approx. 17cm 2 、Approx. 18cm 2 、Approx. 19cm 2 、About 20cm 2 、Approx. 25cm 2 、About 30cm 2 、Approx. 35cm 2 、Approx. 40cm 2 、Approx. 45cm 2 、Approx. 50cm 2 、Approx. 55cm 2 、Approx. 60cm 2 、Approx. 65cm 2 、Approx. 70cm 2 、Approx. 75cm 2 、Approx. 80cm 2 、Approx. 85cm 2 、Approx. 90cm 2 、Approx. 95cm 2 , or about 100cm 2 .

10. The method according to any one of the preceding claims, characterized in that The affected area of ​​the subject was the skin.

11. The method according to any one of the preceding claims, characterized in that The subject's affected area is located at one or more locations independently selected from the group consisting of: scalp, forehead, forearm, face, nose, ear, eyelid, lip, neck, arm, hand, trunk, leg, and foot.

12. The method according to any one of the preceding claims, characterized in that The 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is prepared into a pharmaceutical composition suitable for external use.

13. The method according to any one of the preceding claims, characterized in that Based on the total weight of the pharmaceutical composition, the content of the 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is about 0.025-4% by weight, preferably about 0.05-3% by weight, preferably about 0.2-0.8% by weight, preferably about 0.075% by weight, about 0.1% by weight, about 0.125% by weight, about 0.15% by weight, about 0.175% by weight, about 0.2% by weight, about 0.225% by weight, about 0.25% by weight, about 0.275% by weight, about 0.3% by weight, about 0.325% by weight, about 0.35% by weight, about 0.375% by weight, about 0.4% by weight, about 0.425% by weight, about 0.5% by weight, about 0.6% by weight, about 0.7% by weight, about 0.8% by weight, about 0.9% by weight, about 1.1% by weight, about 1.2% by weight, about 1.3% by weight, about 1.4% by weight, about 1.5% by weight, about 1.6% by weight, about 1.7% by weight, about 1.8% by weight, about 1.9% by weight 45 weight%, about 0.475 weight%, about 0.5 weight%, about 0.525 weight%, about 0.55 weight%, about 0.575 weight%, about 0.6 weight%, about 0.625 weight%, about 0.65 weight%, about 0.675 weight%, about 0.7 weight%, about 0.725 weight%, about 0.75 weight%, about 0.775 weight%, about 0.8 weight%, about 0.825 weight%, about 0.85 weight%, about 0.875 weight%, about 0.9 weight%, about 0.925 weight%, about 0.95 weight%, about 0.975 weight%, about 1 weight%, about 1.25 weight%, about 1.5 weight%, about 1.75 weight%, about 2.0 weight%, about 2.25 weight%, about 2.5 weight%, about 2.75 weight%.

14. The method according to any one of the preceding claims, characterized in that The amount of the pharmaceutical composition applied to the affected skin area is about 0.001 g / cm 2 Skin surface area: about 0.5 g / cm 2 The skin surface area is preferably about 0.0015 g / cm 2 Skin surface area: about 0.2 g / cm 2 Skin surface area; preferably about 0.0015 g / cm 2 Skin surface area: about 0.15 g / cm 2 Skin surface area; preferably about 0.002 g / cm 2 Skin surface area: approximately 0.125 g / cm 2 Skin surface area; preferably about 0.003 g / cm 2 Skin surface area: about 0.1 g / cm 2 Skin surface area; preferably about 0.004 g / cm 2 Skin surface area: about 0.005g / cm 2 Skin surface area: about 0.006g / cm 2 Skin surface area: about 0.007 g / cm 2 Skin surface area: about 0.008 g / cm 2 Skin surface area: about 0.009 g / cm 2 Skin surface area: about 0.01g / cm 2 Skin surface area: about 0.0125 g / cm 2 Skin surface area: about 0.015 g / cm 2 Skin surface area: about 0.0175 g / cm 2 Skin surface area: about 0.02g / cm 2 Skin surface area: about 0.03g / cm 2 Skin surface area: about 0.04g / cm 2 Skin surface area: about 0.05g / cm 2 Skin surface area: about 0.06g / cm 2 Skin surface area: about 0.07g / cm 2 Skin surface area: about 0.08g / cm 2 Skin surface area: about 0.09g / cm 2 Skin surface area.

15. The method according to any one of the preceding claims, characterized in that 17β-xanthoside B or its pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug is administered every day, every other day, every two days, every three days, every four days, every five days, every six days, or every seven days; the administration day is administered once, twice, three times, or four times.

16. The method according to any one of the preceding claims, characterized in that The pharmaceutical composition is a gel.

17. The method according to any one of the preceding claims, characterized in that The pharmaceutical composition comprises a gel matrix, wherein the gel matrix is ​​preferably at least one of sodium carboxymethylcellulose, carbomer, gelatin, and alginate, and the content of the gel matrix is ​​about 0.5-5% by weight, preferably about 1-4% by weight, preferably about 1-3% by weight, preferably about 1.5% by weight, about 2% by weight, about 2.5% by weight, about 3% by weight based on the total weight of the pharmaceutical composition; and / or The pharmaceutical composition comprises a humectant, wherein the humectant is at least one selected from alcohol humectants; preferably, the humectant is glycerol, and the content of the humectant is about 5-35% by weight, preferably about 10-30% by weight, preferably about 15-25% by weight, more preferably about 15% by weight, about 20% by weight, about 25% by weight, about 30% by weight, based on the total weight of the pharmaceutical composition; and / or The pharmaceutical composition comprises a solvent; preferably, the solvent is at least one of ethanol, propylene glycol, and benzyl alcohol, more preferably the ethanol is 95% ethanol or anhydrous ethanol; based on the total weight of the pharmaceutical composition, the content of the solvent is about 20-75% by weight, preferably about 25-70% by weight, more preferably about 30% by weight, about 35% by weight, about 40% by weight, about 45% by weight, about 50% by weight, about 55% by weight, about 60% by weight, about 65% by weight; and / or The pharmaceutical composition comprises water, preferably purified water, distilled water or deionized water, and the content of the water is about 20-75% by weight, preferably about 25-70% by weight, more preferably about 30% by weight, about 35% by weight, about 40% by weight, about 45% by weight, about 50% by weight, about 55% by weight, about 60% by weight, about 65% by weight based on the total weight of the pharmaceutical composition; and / or The pharmaceutical composition comprises a permeation enhancer, which is at least one selected from azone, borneol, Tween 80, sodium lauryl sulfate, and poloxamer, more preferably the permeation enhancer is azone or borneol, and the content of the permeation enhancer is about 0-5% by weight based on the total weight of the pharmaceutical composition; preferably about 0.1-4% by weight; more preferably about 0.25% by weight; about 0.5% by weight; about 0.75% by weight; about 1% by weight; about 1.25% by weight; about 1.5% by weight; about 1.75% by weight; about 2% by weight; about 2.25% by weight; about 2.5% by weight; about 2.75% by weight; about 3% by weight; about 3.25% by weight; about 3.5% by weight; about 3.75% by weight.

18. The method according to any one of the preceding claims, characterized in that The pharmaceutical composition comprises a preservative, and the preservative is a paraben preservative and / or butylated hydroxytoluene; preferably, the paraben preservative is methylparaben and / or ethylparaben; based on the total weight of the pharmaceutical composition, the content of the preservative is about 0-4 weight %; preferably about 0.1-3 weight %; more preferably about 0.25 weight %; about 0.5 weight %; about 0.75 weight %; about 1 weight %; about 1.25 weight %; about 1.5 weight %; about 1.75 weight %; about 2 weight %; about 2.25 weight %; about 2.5 weight %; about 2.75 weight %.

19. The method according to any one of the preceding claims, characterized in that The pharmaceutical composition comprises: About 0.2-0.8% by weight of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof; and / or about 1.5% by weight of a gel matrix; and / or about 40% by weight of a solvent; and / or About 0.5% by weight of a penetration enhancer; and / or about 20% by weight of a humectant; and The remaining amount of water.

20. The method according to any one of the preceding claims, characterized in that Administration of 17β-xanthoside B or a pharmaceutically acceptable salt, racemate, stereoisomer, enantiomer, diastereomer, tautomer, metabolite, solvate, prodrug thereof reduces the number and / or severity of local skin reactions or other adverse side effects in a subject compared to other treatments for skin cancer or its precancerous lesions.

21. The method according to any one of the preceding claims, characterized in that The local skin reaction is selected from the group consisting of vesicle formation, pustule formation, erosion, ulceration, redness, swelling, flaking, scaling, lumps, dryness, pus and blisters.

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