Compounds and methods for treating cervical precancerous lesions or skin precancerous lesions

Abstract

This invention provides the use of compounds, their stereoisomers, or pharmaceutically acceptable salts in the preparation of medicaments for the treatment and / or prevention of precancerous cervical lesions or precancerous skin lesions. This invention demonstrates that the compounds can effectively inhibit the activity of cells with precancerous cervical lesions or precancerous skin lesions, and have significant clinical application value.

Description

[0001] This application claims priority to the prior application filed by the applicant with the China National Intellectual Property Administration on April 11, 2025, with patent application number 202510459197.3, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention belongs to the pharmaceutical field, specifically relating to compounds and methods for treating precancerous lesions of the cervix or skin. Background Technology

[0003] Cancer is a malignant proliferative disease, referring to the irreversible carcinogenesis of cells under the influence of genetic and / or environmental factors. This results in cells having the ability to proliferate uncontrollably, invade surrounding tissues, and / or metastasize to distant sites. Its pathological characteristics include significant cellular atypia, active nuclear division, and disordered tissue structure. Currently, cancer has become one of the leading causes of death worldwide. Research has found that the development of precancerous lesions into cancer can take a long time, ranging from several years to decades, providing a sufficient window for intervention. This period is crucial for implementing cancer prevention, early screening, early diagnosis, and early treatment.

[0004] The World Health Organization defines precancerous lesions as those with a greater than 20% chance of developing into cancer. Precancerous lesions specifically refer to benign lesions with a potential risk of becoming cancerous, but which have not yet reached the stage of cancer. However, if they are not treated promptly and effectively, precancerous lesions may develop into cancer. In other words, although the cells in precancerous lesions have shown abnormalities or morphological changes, they have not broken through the basement membrane, lack invasiveness and metastatic ability, and have not yet formed true cancer. Therefore, precancerous lesions are not the same as cancer; they are two different types of lesions. Furthermore, precancerous lesions are not the precursors to cancer, but rather benign lesions with the potential to become cancerous. Precancerous lesions and cancer also differ significantly in their pathological manifestations; they represent different variations, and the degree of variation in precancerous lesions is far less than that in cancer. Their treatment methods also differ. Precancerous lesions may be caused by various factors, such as chronic inflammation, genetic factors, and unhealthy lifestyle habits. With timely and effective treatment, it is possible to completely stop the disease progression and prevent cancer from developing.

[0005] Most cancers have no obvious symptoms in their early stages, and patients are often diagnosed at an advanced stage. At this time, the risk of cancer cell spread and metastasis increases significantly, making treatment more difficult, the prognosis poorer, and patients' quality of life lower, with their lives severely threatened. Therefore, early screening to identify patients with precancerous lesions and taking proactive intervention measures to directly block the path of these patients to cancer has significant clinical significance and value.

[0006] Taking cervical cancer as an example, cervical cancer is one of the most common cancers among women, ranking second among female cancers nationwide, second only to breast cancer. Cervical cancer has become a significant disease threatening women's lives and health, and is a major public health problem that urgently needs to be addressed. The overall 5-year survival rate for cervical cancer is approximately 55%. Early-stage cervical cancer often has no obvious symptoms, and most cases are diagnosed at an advanced stage, resulting in poor treatment outcomes and prognosis. However, the occurrence and development of cervical cancer involves a gradual evolutionary process, which can take anywhere from several years to decades. This evolutionary process is generally considered to go through several stages: mild, moderate, and severe cervical intraepithelial neoplasia (CIN), early invasive carcinoma, and invasive carcinoma. CIN can be divided into three stages: mild intraepithelial neoplasia (CIN I), moderate intraepithelial neoplasia (CIN II), and severe intraepithelial neoplasia (CIN III). Carcinoma in situ is classified as CIN III. The 2006 ASCCP evidence-based medicine consensus guidelines considered CIN II and CIN III to be high-grade precancerous lesions of the cervix that should be treated. Early treatment at the precancerous stage can potentially prevent cervical cancer and improve its cure and survival rates. However, there are currently no drugs for treating precancerous cervical lesions; treatment is generally surgical, which is invasive and prone to recurrence.

[0007] As medical research deepens, researchers are paying more and more attention to the screening and treatment of precancerous lesions. However, due to their differences from cancer, there is still a lack of sufficiently effective treatment drugs and methods for precancerous lesions. Summary of the Invention

[0008] To address the problems existing in the prior art, in a first aspect, the present invention provides the use of the following compounds, their stereoisomers, or pharmaceutically acceptable salts in the preparation of medicaments for treating and / or preventing precancerous cervical lesions or precancerous skin lesions: , , , , , , , , , , , , , , , , .

[0009] According to an embodiment of the present invention, the compound is selected from the following structures:

[0010]

[0011]

[0012] According to an embodiment of the present invention, the precancerous lesions of the cervix include: cervical squamous cell precancerous lesions and cervical glandular cell precancerous lesions. According to an embodiment of the present invention, squamous cell precancerous lesions include: cervical intraepithelial neoplasia grade 1 (CIN 1, CIN I), cervical intraepithelial neoplasia grade 2 (CIN 2, CIN II), and cervical intraepithelial neoplasia grade 3 (CIN 3, CIN III).

[0013] According to embodiments of the present invention, the precancerous lesions of the cervix can also be defined as: low-grade squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL). According to embodiments of the present invention, the precancerous lesion of cervical adenocarcinoma is cervical adenocarcinoma in situ (AIS).

[0014] According to an embodiment of the present invention, the precancerous lesion of the skin includes Bowen's disease.

[0015] According to an embodiment of the present invention, in the use described, the medicament is a composition comprising the compound described in the first aspect, its stereoisomers, or pharmaceutically acceptable salts; According to embodiments of the present invention, the composition may optionally also include one, two or more other therapeutic agents; the therapeutic agents may be used to treat and / or prevent cervical precancerous lesions or skin precancerous lesions.

[0016] In a second aspect, the present invention also provides a pharmaceutical composition for treating and / or preventing precancerous lesions of the cervix or precancerous lesions of the skin, said pharmaceutical composition comprising the compound described in the first aspect, its stereoisomers, or pharmaceutically acceptable salts.

[0017] According to embodiments of the present invention, the pharmaceutical composition may optionally further comprise one, two or more other active agents; According to embodiments of the present invention, the pharmaceutical composition may optionally further comprise other drugs for treating and / or preventing precancerous cervical lesions or precancerous skin lesions.

[0018] According to an embodiment of the present invention, the cervical precancerous lesion or skin precancerous lesion has the foregoing definition.

[0019] Thirdly, the present invention also provides a method for treating and / or preventing precancerous lesions of the cervix or skin, the method comprising administering to a subject a therapeutically effective amount of the compound of the first aspect, its stereoisomer or pharmaceutically acceptable salt, or the pharmaceutical composition of the second aspect.

[0020] According to an embodiment of the present invention, the cervical precancerous lesion or skin precancerous lesion has the foregoing definition.

[0021] Terminology Definitions and Explanations Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures should be understood as being within the scope of this application specification and / or claims.

[0022] The stereochemical definitions and rules used in this invention generally follow those of S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereo chemistry of Organic Compounds,” John Wiley & Sons, Inc., New York, 1994.

[0023] "Stereoisomers" are compounds that have the same chemical structure but whose atoms or groups are arranged differently in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and hindered isomers, etc.

[0024] "Enantiomers" refer to two non-overlapping but mirror-image isomers of a compound.

[0025] A diastereomer is a stereoisomer that has two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and reactivity. Mixtures of diastereomers can be separated by high-resolution analytical procedures such as electrophoresis and chromatography, for example, HPLC.

[0026] Any asymmetric atom (e.g., carbon, etc.) in the compounds disclosed in this invention can exist in racemic or enantiomerically enriched forms, such as (R)-, (S)-, or (R,S)- configurations. In some embodiments, each asymmetric atom has at least 0% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R)- or (S)- configuration.

[0027] Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0028] In the case of racemic amines, diastereomers are obtained from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid in R and S forms, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline), or various optically active camphorsulfonic acids. Chromatographic enantiomer separation can also be advantageously performed using optically active resolving agents (e.g., dinitrobenzoylphenylglycine immobilized on silica gel, cellulose triacetate or other carbohydrate derivatives, or chiral derivatized isobutylene ester polymers). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, such as hexane / isopropanol / acetonitrile.

[0029] The term "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable when administered to humans and generally do not produce allergic or similar inappropriate reactions, such as gastrointestinal upset, dizziness, etc.

[0030] Pharmaceutically acceptable salts can be, for example, acid addition salts of compounds of the present invention having sufficient basicity and containing a nitrogen atom in the chain or ring, such as acid addition salts formed with inorganic acids including: hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfate; or acid addition salts formed with organic acids including: formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, and diglucose. Gluconic acid, 3-hydroxy-2-naphtholic acid, nicotinic acid, pyruvic acid, pectinic acid, persulfate, 3-phenylpropionic acid, picric acid, pentyl acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucohepanoic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, hemisulfonic acid or thiocyanate.

[0031] Alternatively, another suitable pharmaceutically acceptable salt of the compounds of the present invention having sufficient acidity is an alkali metal salt (e.g., sodium or potassium salt), an alkaline earth metal salt (e.g., calcium or magnesium salt), an ammonium salt, or a salt formed with an organic base that provides a physiologically acceptable cation, such as a salt formed with sodium ions, potassium ions, N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serine, trihydroxymethylaminomethane, aminopropylene glycol, 1-amino-2,3,4-butanetriol. As an example, the pharmaceutically acceptable salts include salts formed by the -COOH group with the following substances: sodium ion, potassium ion, calcium ion, magnesium ion, N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serine, trihydroxymethylaminomethane, aminopropylene glycol, and 1-amino-2,3,4-butanetriol.

[0032] In addition, basic nitrogen-containing groups can be quaternized using the following reagents: lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and dipentyl sulfate; long-chain halides, such as decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides; and aralkyl halides such as benzyl and phenethyl bromides. As examples, pharmaceutically acceptable salts include hydrochlorides, sulfates, nitrates, hydrogen sulfates, hydrobromides, acetates, oxalates, citrates, methanesulfonates, formates, or meglumine salts.

[0033] Since the compounds of the present invention can have multiple salt-forming sites, the pharmaceutically acceptable salt includes not only the salt formed at one of the salt-forming sites of the compounds of the present invention, but also the salt formed at two, three, or all of the salt-forming sites. Therefore, the molar ratio of the compound of formula (I) to the anion of the acid or the cation of the base required for salt formation in the pharmaceutically acceptable salt can vary over a wide range, for example, it can be 4:1 to 1:4, such as 3:1, 2:1, 1:1, 1:2, 1:3, etc.

[0034] In this invention, "solvent" refers to an association formed by one or more solvent molecules with the compound of this invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed when the solvent molecules are water.

[0035] As used in this invention, the term "treatment" refers to any disease or condition, and in some embodiments, it means improving the disease or condition (i.e., slowing down or stopping or alleviating the development of the disease or at least one of its clinical symptoms). In other embodiments, "treatment" means alleviating or improving at least one bodily parameter, including bodily parameters that may not be perceived by the patient. In still other embodiments, "treatment" means regulating the disease or condition physically (e.g., stabilizing perceptible symptoms) or physiologically (e.g., stabilizing bodily parameters) or both. In still other embodiments, "treatment" means preventing or delaying the onset, occurrence, or worsening of the disease or condition.

[0036] The term "effective amount" or "therapeutic effective amount" refers to the amount of the compound of this invention that is sufficient to achieve the intended application (including, but not limited to, the treatment of diseases as defined below) without toxicity or acceptable toxicity. Therapeutic effective amounts may vary depending on factors such as the intended application (in vitro or in vivo), or the subject being treated and the condition of the disease, such as the subject's weight and age, the severity of the disease, and the route of administration, which can be readily determined by those skilled in the art. Specific dosages will vary depending on factors such as the specific compound selected, the dosing regimen, whether it is administered in combination with other compounds, the timing of administration, the tissue to which the drug is administered, and the physical delivery system used.

[0037] Unless otherwise stated, any abbreviations for protecting groups, amino acids and other compounds used in this invention shall be those that are commonly used and recognized, or refer to the IUPAC-IUB Commission on Biochemical Nomenclature (see Biochem. 1972, 11: 942-944).

[0038] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0039] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention. Attached Figure Description

[0040] Figure 1 The number of live cells in CIN3 precancerous cervical lesions was determined (n=3, *P<0.05, **P<0.01, ***P<0.001). Detailed Implementation

[0041] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0042] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0043] Example 1: Basic structural information of the active compound 17β-Xanthin (compound EEO) has the following structure: .

[0044] 17β-Xanthocyanin derivatives: compounds EE1 to EE17. The preparation process of compounds EE1 to EE17 can be found in patent CN202510289442.0.

[0045] Chemical name of compound EE1: (2R,3S,4R,5S,6S)-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran -3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3,5-diyl dimethylbis(carbonate).

[0046] Chemical name of compound EE2: (2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrof uran-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-ylmethyl carbonate.

[0047] The chemical name of compound EE3: diethyl ((2R,3S,4R,5S,6S)-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran -3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3,5-diyl) bis(carbonate).

[0048] Chemical name of compound EE4: ethyl ((2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihyd rofuran-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-yl) carbonate.

[0049] The chemical name of compound EE5: (2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrof uran-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-ylpropyl carbonate.

[0050] The chemical name of compound EE6: (2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofur an-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-ylisopropyl carbonate.

[0051] The chemical name of compound EE7: (2R,3S,4R,5S,6S)-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran -3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3,5-diyl dipropylbis(carbonate).

[0052] The chemical name of compound EE8: (2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran- 3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-yl(2-hydroxyethyl) carbonate.

[0053] The chemical name of compound EE9: (2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran-3 -yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-yl(3-hydroxypropyl) carbonate.

[0054] The chemical name of compound EE10: (2S,3S,4S,5S,6R)-5-hydroxy-6-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrof uran-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-2-methyltetrahydro-2H-pyran-3-ylpropyl carbonate.

[0055] Chemical name of compound EE11: hexyl ((2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihyd rofuran-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-yl) carbonate.

[0056] The chemical name of compound EE12: (2R,3S,4R,5S,6S)-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran-3-yl) hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3,5-diyldiisopropyl bis(carbonate).

[0057] The chemical name of compound EE13: (2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran-3-yl )hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-yl(1-methylpiperidin-4-yl) carbonate.

[0058] The chemical name of compound EE14: (2R,3S,4R,5S,6S)-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran -3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3,5-diyl bis(1-methylpiperidin-4-yl) bis(carbonate).

[0059] The chemical name of compound EE15: 1-ethylpiperidin-4-yl ((2R,3S,4R,5S,6S)-5-hydroxy-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihyd rofuran-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3-yl) carbonate.

[0060] The chemical name of compound EE16: (2R,3S,4R,5S,6S)-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran -3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3,5-diyl bis(3-hydroxypropyl) bis(carbonate).

[0061] The chemical name of compound EE17: (2R,3S,4R,5S,6S)-2-(((3S,5R,8R,9S,10S,13R,14S,17R)-14-hydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran -3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-4-methoxy-6-methyltetrahydro-2H-pyran-3,5-diyl bis(2-hydroxyethyl) bis(carbonate).

[0062] The structural formulas of compounds EE1 to EE17 are shown in Table 1.

[0063] Table 1. Structural formulas of compounds EE1 to EE17

[0064]

[0065]

[0066] Example 2: Study on the in vivo antitumor activity of the compound Using a human cervical cancer HeLa cell Balb / c-nu mouse model, in vivo activity screening experiments were conducted on different 17β-flavin derivatives by intramuscular injection and oral administration.

[0067] Tumor measurement method: Hold the mouse with your left hand to fix it, and then use a vernier caliper with your right hand to measure and record the longest diameter (a) and short diameter (b) of the tumor.

[0068] The formula for calculating tumor volume (TV) is: V = 1 / 2 × a × b 2 .

[0069] The relative tumor volume (RTV) is calculated based on the measurement results using the formula: RTV = Vt / V0. Where V0 is the tumor volume measured during individual drug administration (i.e., d0), and Vt is the tumor volume at each measurement.

[0070] Calculate the relative tumor proliferation rate T / C (%) using the following formula: T / C (%) = T RTV / C RTV ×100%. (T) RTV Treatment group RTV; C RTV(Model control group RTV).

[0071] Experimental results showed that the antitumor activity of the compound of this invention was significantly better than that of other types of derivatives.

[0072] The antitumor activities of the representative compounds of this invention are as follows: EE5: QD (once daily) oral administration of 1.6 mg / Kg, relative tumor proliferation rate at the end of administration D21 (day 21): 39.9%.

[0073] Other representative derivatives: MW794.89 (LCMS, M+1, 795.37); QD (once daily) oral 2.1 mg / kg, Q3D (every three days) intramuscular injection 2.1 mg / kg, both were inactive (the relative tumor proliferation rate at D16 after the end of administration was greater than 99%).

[0074] Example 3 Pharmacodynamic Experiment - Study on the Therapeutic Effect of Compounds on Precancerous Lesions 1. Effect of compound EE7 on the number of viable cells in clinical tissues of precancerous cervical lesions The effect of compound EE7 on the number of viable cells in clinical tissues with CIN3 precancerous cervical lesions was investigated using the CCK8 assay.

[0075] 1.1 Test Methods (1) Group setup: A negative control group, compound EE7 groups (200 ng / mL, 600 ng / mL, 2000 ng / mL and 6000 ng / mL groups), and a positive control Cisplatin group (1 μM, 3 μM and 10 μM groups) were set up. The solvent used in each group was 0.5% DMSO + 0.5% ethanol at the concentration of the culture medium. The negative control group consisted of clinical tissues of cervical precancerous lesions treated with solvent only; the compound EE7 group consisted of clinical tissues of cervical precancerous lesions treated with compound EE7 at concentrations of 200 ng / mL, 600 ng / mL, 2000 ng / mL and 6000 ng / mL, respectively; the positive control Cisplatin group consisted of clinical tissues of cervical precancerous lesions treated with cisplatin at concentrations of 1 μM, 3 μM and 10 μM, respectively; each group had three replicates.

[0076] (2) In each well of a 96-well plate, add 100 μL of complete culture medium containing 20% ​​FBS containing the test substance, and then add 100 μL of clinical samples of cervical precancerous lesions to each well. Incubate at 37°C and 5% CO2 for 144 h.

[0077] (3) CCK8 assay: After culturing for 144 h, the number of viable cells in the tissue was detected using the CCK8 assay. 20 μL of CCK8 reagent was added to each well, and the well was incubated at 37°C for 2 h. The well was then shaken for 20 s on a microplate reader, and the OD value was detected at a wavelength of 450 nm. The viable cell rate (Live cell number %) was calculated using the following formula: Live cell number % = (OD of compound group – zeroing OD) / (OD of negative control group – zeroing OD) * 100%; The inhibition rate (IR) of the detected tissue can also be further calculated using the following formula: IR (%) = 100% - Live cell number %.

[0078] 1.2 Test Results The CCK8 test results are shown in Table 2 and... Figure 1 Compared with the negative control group, as the concentration of compound EE7 increased, the viable cell rate of clinical tissues with CIN3 cervical precancerous lesions decreased and the viable cell inhibition rate increased, with the highest inhibition rate being 92.1% (6000 ng / ml).

[0079] Table 2 CCK8 test results

[0080] Based on the above experimental indicators, it can be seen that the MCM complex inhibitor of the present invention has a significant inhibitory effect on the number of viable cells in clinical tissues of precancerous cervical lesions.

[0081] Example 4: Compound's effect on vaginal stimulation in SD rats 1. To detect the irritant effect and intensity of the compound on the vagina of experimental animals.

[0082] 1.1 Test Methods Healthy, early-adult female SD rats, weighing 175-200 g, were selected. Before the experiment, the vaginal opening of the rats should be checked for discharge, congestion, edema, and other injuries. If inflammation and / or injury were found, the rats should be discarded. The experiment was conducted on rats outside of estrus. The experiment consisted of a treatment group and a control group, with three rats in each group.

[0083] Rats in all groups were administered the drug vaginally once daily for 7 days (each compound was administered in equimolar amounts). The experimental drugs were prepared from the test compound and 5% stearic acid / polyethylene glycol 400 / polyethylene glycol 4000 and 94.5% mixed fatty acid glycerides. During the administration period, the general condition of the rats was observed and they were weighed daily. Photographs of the rat vaginal opening were taken on days 0, 3, and 7 after administration. After the administration period, the rats were anesthetized and necropsies were performed to observe for damage to major organs and the vagina-uterus-ovary system. The vagina-uterus-ovary system was then removed for photographic recording, vaginal weight was measured, and vaginal edema was assessed.

[0084] Table 3. Summary of Groups and Dosage Design

[0085] Note: pv: vaginal administration; QD×7: once daily for 7 consecutive days.

[0086] 1.2 Observation and indicator monitoring: 1.2.1 General observation and vaginal opening photography Adaptation period: Observe the general condition of the rats and whether there is any discharge, congestion, edema and other damage at the vaginal opening.

[0087] During the daily administration period, the experimental rats were observed and their data recorded. Observations included: the presence or absence of vaginal discharge, congestion, edema, and other injuries at the vaginal opening; whether the rats were dead or near death; their food and water intake; presence of external injuries; feces; appearance; fur; mental state; and activity level. Photographs of the rat vaginal opening were taken on days 0, 3, and 7.

[0088] 1.2.2 Weight Adaptation period: Rats were weighed before receiving and grouping.

[0089] Dosing period: During the dosing period, the experimental rats were weighed and their weight recorded daily. After the last dose, the rats were anesthetized and euthanized. Dissection was performed to observe the uterus, ovaries, and major organs for any visible abnormalities, and photographs were taken. The uterus was dissected from the fundus down to the vaginal opening, and the intact ovary-uterus-vaginal region was removed. After rinsing with physiological saline, the vaginal mucosa was observed for congestion, edema, degeneration, etc., and photographs were taken. Vaginal weight and vaginal edema scores were recorded (scoring criteria are shown in Table 5). After fixation with 4% paraformaldehyde, gross necropsy was performed to observe the major organs and tissues: heart, liver, spleen, lungs, kidneys, stomach, and colon and rectum. If any visible lesions were found, pathological examination was required.

[0090] Table 4. Scoring criteria for vaginal edema

[0091] 1.3 Test Results 1.3.1 General observation and vaginal opening photography observation No secretions, congestion, edema, or other damage were found at the vaginal opening of rats during the adaptation period.

[0092] During the drug administration period, no secretions, congestion, edema or other damage were found at the vaginal opening of the rats, and their mental state and activity were normal. No abnormalities were observed.

[0093] 1.3.2 Weight As can be seen from the table below, there were no significant differences in the body weight of the animals in each group at different time points, indicating that the compound of this application had no effect on the body weight of the animals under the experimental conditions.

[0094] Table 5. Body weight of animals in each group at different time points (n=3, )

[0095] 1.3.3 Final necropsy observation, vaginal weight, and edema score: Table 6. Vaginal weight and vaginal edema scores of each group after the drug administration period (n=3, )

[0096] *P<0.05 compared with the control group. After the drug administration period ended, the animals in each group were anesthetized and subjected to necropsy. The experimental results showed that the EE7 group did not cause any visible pathological changes in the major organs of SD rats. In contrast, the EE0 group showed a significant increase in vaginal weight (P<0.05 vs. control group), and mild vaginal edema was visible to the naked eye. Furthermore, the vaginal edema score showed that the EE7 and EE8 groups had a vaginal edema irritation score of 0, which was significantly better than the EE0 and EE9 groups.

[0097] Example 5: Study on the therapeutic effect of compounds on Bowen's disease This clinical study was conducted to evaluate the activity and safety of 17β-xanthinylglycoside (compound EEO). Results showed that 17β-xanthinylglycoside had good clinically relevant activity in the treatment of Bowen's disease; and it was safe based on local tolerability and local skin response (LSR).

[0098] 5.1 Observation Indicators CR: Complete remission, indicating that all tumor target lesions have disappeared; PR: Partial remission, indicating that the tumor volume has shrunk by at least 30%; SD: The disease is stable, indicating that there is no significant change in tumor size; Objective response rate (ORR): The proportion of subjects with CR and PR (calculated based on the best response during the dosing period) to the corresponding analysis set of subjects; Disease control rate (DCR): The proportion of subjects with CR, PR, and SD (calculated based on the best response during the dosing period) out of the subjects in the corresponding analysis set; Duration of remission (DOR): refers to the time from the first assessment of CR or PR to the first assessment of PD (Progressive Disease) or death from any cause. Time to progression of tumor (TTP): refers to the time elapsed from the first time a subject takes medication until the date of confirmed disease progression; Progression-free survival (PFS): refers to the time elapsed from the first dose of medication to the date of definite disease progression or death from any cause. 5.2 Validity Results The efficacy of 17β-xanthin ethinyl glycoside in the treatment of Bowen's disease This study enrolled 6 patients with evaluable Bowen's disease in the first phase of a clinical trial. Based on investigator assessment (according to RECIST 1.1 criteria), the disease control rate (DCR) of 17β-flavin glycoside B in treating Bowen's disease was 100%; the complete response rate (CRR) was 16.67%; and the objective response rate (ORR) was 66.7%.

[0099] Example 6: Toxicity / Safety Study In this experiment, Balb / c mice were randomly divided into groups of three based on body weight and administered the compounds of this invention. The mice were administered the compounds via gavage or intravenous injection, respectively. The gavage dose was 5 mg / kg of each test substance, with a volume of 5 mL / kg. The intravenous injection dose was 3 mg / kg of each test substance, with a volume of 3 mL / kg. The toxic effects on the mice were observed after administration. The results are as follows: No significant abnormal reactions were observed in EE7 when administered via gavage at a dose of 5 mg / kg or via intravenous injection at a dose of 3 mg / kg; however, EE9 exhibited abnormalities such as bradykinesia or prone position after administration of EE7 via gavage at a dose of 5 mg / kg or via intravenous injection at a dose of 3 mg / kg.

[0100] The above results indicate that the compound EE7 of this invention has relatively superior safety.

[0101] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Use of a compound, its stereoisomer, or a pharmaceutically acceptable salt in the preparation of a medicament for treating precancerous cervical lesions or precancerous skin lesions, said compound being selected from: 、 、 , , , , , , , , , , , , , ,or .

2. The use according to claim 1, characterized in that, The compound is selected from the following structures: 。 3. The use according to claim 1 or 2, characterized in that, The drug may optionally include one, two or more other therapeutic agents; the therapeutic agents may be used to treat and / or prevent cervical precancerous lesions or skin precancerous lesions.

4. The use according to any one of claims 1-4, characterized in that, The precancerous lesions of the cervix include: cervical squamous cell precancerous lesions and cervical glandular cell precancerous lesions; preferably, squamous cell precancerous lesions include: cervical intraepithelial neoplasia grade 1 (CIN 1, CIN I), cervical intraepithelial neoplasia grade 2 (CIN 2, CIN II), and cervical intraepithelial neoplasia grade 3 (CIN 3, CIN III); preferably, the precancerous lesions of the cervix are cervical intraepithelial lesions, including low-grade squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL). Preferably, the precancerous lesion of cervical adenocarcinoma is cervical adenocarcinoma in situ (AIS).

5. The use according to any one of claims 1-4, characterized in that, The precancerous skin lesions mentioned are selected from Bowen disease.

6. A pharmaceutical composition for treating and / or preventing precancerous lesions of the cervix or skin, said pharmaceutical composition comprising the compound of claim 1 or 2, its stereoisomer, or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition according to claim 6, characterized in that, The pharmaceutical composition may optionally further include other medications for treating and / or preventing precancerous cervical lesions.

8. A method for treating and / or preventing precancerous lesions of the cervix or skin, the method comprising administering to a subject a therapeutically effective amount of the compound of claim 1 or 2, its stereoisomer, or a pharmaceutically acceptable salt thereof.

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